AU2015268754B2 - Suppression of cancer - Google Patents

Abstract

Abstract The present invention relates to polypeptides for use in suppressing cancer and cancer disorders. The treatment employs use of a non-cytotoxic protease, which is targeted to the cancer cell, and, when so delivered, the protease is internalised and inhibits 5 secretion from the cancer cell.

Description

The present invention relates to polypeptides for use in suppressing cancer and cancer disorders. The treatment employs use of a non-cytotoxic protease, which is targeted to the cancer cell, and, when so delivered, the protease is internalised and inhibits secretion from the cancer cell.

2015268754 16 Dec 2015

Australia

Patents Act 1990

COMPLETE SPECIFICATION STANDARD PATENT

Invention Title:

Suppression of cancer

P/00/011 Regulation 3.2

The following statement is a full description of this invention, including the best method of performing it known to us:

1001301673

2015268754 16 Dec 2015

Suppression of cancer

Cross reference to related applications

This application is a divisional of Australian patent application no. 2009315435, the entire disclosure of which is incorporated herein by reference.

The present invention relates to the suppression of cancer.

Cancers are made up of cells that divide, invade and survive in an aberrant manner in the body. Cancers develop when alterations to the DNA within cells affect the control of cell division and other processes relevant to cell survival. As cancerous tumours grow, they invade the body tissues surrounding them. This is harmful to the body as it damages surrounding normal tissues and has a significant effect on physiological responses. In addition, cancers may spread to other parts of the body and develop into secondary tumours. Assessment of cancer can be undertaken using a variety of tests including CT scan, MRI imaging, PET scans, combined PET-CT scan and ultrasound tests. Generally, the earlier a cancer is detected and confirmed, the better the overall prognosis for the patient.

A wide range of treatments are available for patients diagnosed with cancer. These include a variety of surgical interventions, radiotherapy, chemotherapy and hormone therapies. These treatments may be used in isolation but most commonly they are used in combination with each other. The use of a combination of therapies is dependent on the type of cancer diagnosed with certain cancers (such as breast cancer) commonly requiring multiple therapeutic interventions. There are a variety of new therapeutic approaches under development utilising biological products including antibodies, immunotherapies and vaccination techniques to treat cancer. Additional treatment approaches including the use of novel therapeutics designed to block angiogenesis are under development. Novel approaches including the use of gene therapy are also being investigated as potential therapeutics. However, despite the significant scientific efforts and huge financial commitment to developing new cancer therapies, the prognosis for many patients remains extremely poor.

Breast cancer is the most common cancer in the UK and every year more than 44,000 women are diagnosed with this type of cancer. Worldwide, more

-1A2015268754 16 Dec 2015 than a million women are diagnosed with breast cancer every year. In the UK, approximately 80% of patients will survive for 5 years or longer.

Lung cancer is the 2nd commonest cancer in the UK and over 37,000 people 5 were diagnosed with lung cancer in the UK in 2003. For patients diagnosed with lung cancer, only approximately 20% will survive for at least 1 year after diagnosis and only approximately 6% will live for 5 years or longer after diagnosis.

Colorectal cancer is the 3rd commonest cancer type overall and it is the 2nd commonest affecting women (after breast cancer). Just over 35,000 people were diagnosed with colorectal cancer in the UK in 2003. Of those diagnosed with bowel and rectal cancer in England and Wales, 46% will survive for at least 5 years after their diagnosis.

Pancreatic cancer is the fourth most common cause of death from cancer in the Western world and over 7000 cases are diagnosed in the UK each year. Despite therapeutic advances only 3-4% of those diagnosed with pancreatic cancer survive for 5 years or longer and successful therapeutic intervention is limited to surgery that is applicable to only 15% of patients.

Renal cancer is the 9th commonest cancer type overall with over 7000 new cases diagnosed and greater than 3500 deaths in the UK each year. Around 208,500 new cases of renal cancer are diagnosed in the world each year, accounting for just under 2% of all cancers. It is commoner in men than in women, with a male to female ratio of 1.5:1. Thirty percent of renal cancer patients show signs of advanced renal cell carcinoma at initial diagnosis, with metastases detected in 15- 25% of patients at this time.

Cancer represents a worldwide problem that is predicted to grow significantly. In the year 2000, malignant tumours were responsible for 12 per cent of the nearly 56 million deaths reported worldwide from all causes. In some countries, more than a quarter of deaths could be attributed to cancer. In 2000, 5.3 million men and 4.7 million women developed a malignant tumour

2015268754 19 Dec 2017

A and altogether 6.2 million died from the disease. It is predicted that cancer rates could further increase by 50% to 15 million new cases in the year 2020, (WHO World Cancer Report (IARC, Ed B.W. Stewart and P. Kleihues, 2003).

For a large number of cancers, autocrine signalling (defined as a mode of hormone action in which a hormone binds to receptors on and affects the function of the cell type that produced it) and/ or paracrine signalling (defined as a mode of hormone action in which hormone released from endocrine or endocrine-like cells binds to receptors on nearby cells and affects their function), are believed to play a significant role in development of the disease state. For a given cancer, multiple autocrine signalling loops may be implicated in development and maintenance of the cancer state. In addition to providing a stimulus for cell division, autocrine signalling can enhance cell survival acting to protect cells from mechanisms of apoptosis and necrosis and increase the metastatic potential of the cancer cell.

There is therefore a need in the art for new therapies/ therapeutics capable of specifically addressing cancer. This need is addressed by the present invention, which solves one or more of the above-mentioned problems.

In more detail, a first aspect of the present invention provides a method of suppressing or treating cancer by inhibiting autocrine secretion from a cancer cell in a patient, said method comprising administering to the patient an effective amount of a polypeptide comprising (i) a non-cytotoxic protease, which protease is capable of cleaving a SNARE protein in a cancer cell;

(ii) a Targeting Moiety (TM) that is capable of binding to a Binding Site on a cancer cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the cancer cell; and (iii) a translocation domain that is capable of translocating the protease from within an endosome, across the endosomal membrane and into the cytosol of the cancer cell;

3a

2015268754 19 Dec 2017 wherein SNARE protein expression is up-regulated in the cancer cell when compared with SNARE protein expression in the same cell type in a noncancerous state;

wherein the polypeptide lacks the natural binding function of a clostridial 5 neurotoxin H_cc domain which enables the clostridial neurotoxin to bind to nerve terminals at the neuromuscular junction;

and with the proviso that the cancer cell is not a neuroendocrine tumour cell; and with the proviso that the polypeptide is not a clostridial neurotoxin (holotoxin) molecule.

2015268754 16 Dec 2015

The polypeptide of the present invention is not a naturally-occurring clostridial neurotoxin molecule (also known as clostridial holotoxin). Clostridial holotoxin is one of the most lethal neurotoxins known to man, and, as such, has significant limitations as a therapeutic molecule. Also, in the context of cancer, clostridial holotoxin is associated with undesirable off-site targeting (ie. targeting of noncancer cells), for example to the neuromuscular junction.

In use, a polypeptide of the invention binds to a cancer cell. Thereafter, the 10 translocation component effects transport of the protease component into the cytosol of the cancer cell. Finally, once inside, the protease inhibits the exocytic fusion process of the cancer cell by cleaving SNARE protein present in the cytosol of the cancer cell. Thus, by inactivating the exocytic fusion apparatus of the cancer ceil, the polypeptide of the invention inhibits secretion (eg. of TNF15 alpha, acetylcholine, fibroblastic growth factor, gastrin releasing peptide, interleukin-6, VEGF, and/ or autocrine mobility factor) therefrom. Accordingly, where said secretion peptides are, for example, involved in autocrine or paracrine signalling, the polypeptide of the invention reduces the signalling drive that the autocrine/ paracrine loop would otherwise provide to the cancer cell. Accordingly, the polypeptides of the present invention are capable of suppressing or treating cancer.

The first aspect also embraces a corresponding method for suppressing cancer, said method comprising administering a therapeutically effective amount of a polypeptide of the present invention to a patient. The present invention thereby provides an effective means for treating cancer such as renal, breast, lung, pancreatic, and colorectal (eg. colon).

The polypeptides of the present invention provide a distinct advantage over other therapeutics in that they have the potential to inhibit the secretion of multiple autocrine and paracrine peptides from a targeted cancer cell using a single therapeutic molecule. This reduces significantly the ability of a cancer to adapt to a specific treatment regime by switching to and using alternative autocrine signalling pathways. Thus, the present invention provides a more

-4 2015268754 16 Dec 2015 efficacious therapy by blocking multiple pathways simultaneously.

The principal target cell of the present invention is a cancer cell, for example a cancer cell that secretes one or more hormones (or other bioactive molecules), which, once secreted, bind to a receptor on and affect the function of said cancer cell. Examples of cancer cells targeted by the present invention include: breast, lung, pancreatic, colorectal (eg. colon), adrenal, oesophageal, lymphoma (eg. B-cell; Mantell cell), leukaemia (eg. multiple myeloma), acute leukaemia, bladder, bone, brain tumours, bowel, cervical, chronic lymphocytic leukaemia,

Hodgkin’s lymphoma, renal, liver, skin (eg. basal, squamous, melanoma), oropharyngeal, myeloma, prostate (eg. soft tissue sarcoma), gastric, testicular, and uterine.

In one embodiment, the target cell of the present invention is a cancer cell in which the level of SNARE protein expression is undesirable.

For example, undesirable SNARE expression may occur when the expression of a SNARE protein is up-regulated when compared with the same (or very similar) cell type when in a non-cancerous state. In one embodiment, the mRNA encoding a SNARE protein is up-regulated in a target cancer cell. In another (or the same) embodiment, the cellular protein concentration of a SNARE protein is (also) increased in the target cancer cell. When measuring SNARE mRNA or protein levels, a value of One’ may be assigned to the SNARE mRNA or protein concentration in a non-cancerous cell of a normal individual, which cell is otherwise the same (or very similar) to the cell type of the cancer cell in question - this value might be described as a ‘basal expression level’, and is typically an average of expression level values obtained from several cells (of the same or very similar type) and/ or from different normal individuals. For example, a lung cell (e.g. a neuroectodermal lung cell, or a lung epithelium cell) in a normal individual may be used for comparison with a small cell lung cancer cell. Thus, up-regulated in the context of the present invention means that the SNARE mRNA or protein level is ‘greater than 1’ when compared with the same cell type in a non-cancerous state in a normal individual. In one embodiment, the SNARE protein or mRNA level is up-regulated by greater than 2- or 3-fold, or by greater

-5 2015268754 16 Dec 2015 than 4- or 5-fold, or by greater than 6- or 7-fold. In another embodiment, the SNARE protein or mRNA level is up-regulated by greater than 8- or 9-fold, or by greater than 10- or 12-fold, or by greater than 15- or 20-fold. In another embodiment, the SNARE protein or mRNA level is up-regulated by greater than

40- or 50-fold, or by greater than 60- or 70-fold, or by greater than 90- or 100fold.

Alternatively, undesirable SNARE expression may occur when the level of SNARE mRNA and/ or protein expression in a cancer cell is substantially normal (ie. substantially the same) when compared with that of a corresponding identical or very similar non-cancerous cell type. In this scenario, however, for example when a patient is suffering from a [prostate, bladder or cervical carcinoma], even a low or normal level of SNARE protein expression may support a secretion from the cancer cells of this type that is paracrine or autocrine in function, and so in the context of the cancer, the SNARE protein is supporting an undesirable secretion. In this scenario, the polypeptides of the present invention are capable of suppressing secretion from said cancer ceils, and thereby addressing this need in the art.

In this regard, the present inventors have identified undesirable SNARE expression in a range of cancer cell types. These data are presented as Box Plots in Figures 3-47. In summary, the present inventors have confirmed that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-1a, syntaxin-2), and VAMP (eg. VAMP-1) is observed in small cell lung cancer cells (eg. when compared with normal lung cells); that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-2), VAMP (eg. VAMP-1, VAMP-2, VAMP-3) is observed in lung carcinoid cells; that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-2), VAMP (eg. VAMP-1, VAMP-2) is observed in lung carcinoid cells (eg. when compared with normal lung cells); that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-2, syntaxin-3), and VAMP (eg. VAMP-1, VAMP-3) is observed in bladder carcinoma cells; that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-1a), and VAMP (eg. VAMP-2) is observed in breast

- 6 2015268754 16 Dec 2015 cancer cells; that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-1a, syntaxin-1b, syntaxin-2, syntaxin-3), and VAMP (eg. VAMP-1, VAMP-2, and VAMP-3) is observed in pancreatic adenocarcinoma cells (eg. when compared with normal pancreatic cells); that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-1a, syntaxin-1b, syntaxin3), and VAMP (eg. VAMP-2, VAMP-3) is observed in prostate carcinoma cells (eg. when compared with benign prostate cells); that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-2), and VAMP (eg. VAMP3) is observed in cervix cancer cells (eg. when compared with normal cervix cells); that undesirable SNARE expression such as syntaxin (eg. syntaxin 3) and VAMP (eg. VAMP-1, VAMP-2, VAMP-3) is observed in B-celi leukaemia cells (eg. when compared with normal B-cell lymphocytes); that undesirable SNARE expression such as syntaxin (eg. syntaxin-1a, syntaxin-3) and VAMP (eg. VAMP-1) is observed (eg. up-regulated) in ovarian cancer cells (eg. when compared with normal ovary cells); that undesirable SNARE expression such as SNAP-25 and VAMP (eg. VAMP-1, VAMP-2, VAMP-3) is observed in gastrointestinal (Gl) carcinoid cells (eg. when compared with normal Gl cells); and that undesirable SNARE expression such as SNAP-25, syntaxin (eg. syntaxin-1a, syntaxin-2) and VAMP (eg. VAMP-3) is observed (eg. up-regulated) in head and neck cancer cells (eg. when compared with normal head and neck cells). In addition, the present inventors have identified that undesirable SNARE expression such as SNAP-25 and VAMP (eg. VAMP-2) is observed in colon cancer cells; that undesirable SNARE expression such as SNAP-25 is observed in adrenal cancer cells; that undesirable SNARE expression such as syntaxin (eg. syntaxin-2) is observed in skin (eg. melanoma) cancer cells; that undesirable SNARE expression such as syntaxin (eg. syntaxin-3) is observed in leukemia (eg. multiple melanoma) cancer cells; that undesirable SNARE expression such as syntaxin (eg. syntaxin-1a) and VAMP (eg. VAMP-1) is observed in lung adenocarcinoma cells; that undesirable SNARE expression such as VAMP (eg. VAMP-1) is observed in liver cancer cells; that undesirable SNARE expression such as VAMP (eg. VAMP-2) is observed in oesophagus cancer cells; and that undesirable SNARE expression such as VAMP (eg. VAMP-3) is observed in lymphoma cancer cells (eg. B-cell lymphoma and Mantell cell).

- 7 2015268754 16 Dec 2015

The ‘bioactive’ component of the polypeptides of the present invention is provided by a non-cytotoxic protease. This distinct group of proteases act by proteolyticaliy-cleaving intracellular transport proteins known as SNARE proteins (e.g. SNAP-25, VAMP, or Syntaxin) - see Gerald K (2002) Cell and Molecular Biology” (4th edition) John Wiley & Sons, Inc. The acronym SNARE derives from the term Soluble NSF Attachment Receptor, where NSF means N-ethylmaleimide-Sensitive Factor. SNARE proteins are integral to intracellular vesicle formation, and thus to secretion of molecules via vesicle transport from a cell. Accordingly, once delivered to a desired target cell, the non-cytotoxic protease is capable of inhibiting cellular secretion from the target cell.

Non-cytotoxic proteases are a discrete class of molecules that do not kill cells;

instead, they act by inhibiting cellular processes other than protein synthesis. Non-cytotoxic proteases are produced as part of a larger toxin molecule by a variety of plants, and by a variety of microorganisms such as Clostridium sp. and Neisseria sp.

Clostridial neurotoxins represent a major group of non-cytotoxic toxin molecules, and comprise two polypeptide chains joined together by a disulphide bond. The two chains are termed the heavy chain (H-chain), which has a molecular mass of approximately 100 kDa, and the light chain (L-chain), which has a molecular mass of approximately 50 kDa. It is the L-chain, which possesses a protease function and exhibits a high substrate specificity for vesicle and/or plasma membrane associated (SNARE) proteins involved in the exocytic process (eg. synaptobrevin, syntaxin or SNAP-25). These substrates are important components of the neurosecretory machinery.

Neisseria sp., most importantly from the species N. gonorrhoeae, and Streptococcus sp., most importantly from the species S. pneumoniae, produce functionally similar non-cytotoxic toxin molecules. An example of such a noncytotoxic protease is IgA protease (see WO99/58571, which is hereby incorporated in its entirety by reference thereto). Thus, the non-cytotoxic

- 8 2015268754 16 Dec 2015 protease of the present invention is preferably a clostridial neurotoxin protease or an IgA protease.

Turning now to the Targeting Moiety (TM) component of the present invention, it 5 is this component that binds the polypeptide of the present invention to a cancer cell. The TM is preferably a peptide.

The TM binds to a Binding Site on the cancer cell, thereby providing selectivity of the polypeptide to this species of target cell over other cells. By way of example, in gastric cancer, the TM preferably binds to gastric cancer cells in preference to other cells of the stomach, such as those that are not cancerous and/ or to other non-cancerous (or cancerous) cells in the body. In this regard, preferred TM embodiments of the present invention include antibodies (eg. monoclonal antibodies, antibody fragments such as Fab, F(ab)’2, Fv, ScFv, etc., and antibody domains peptides), as well as binding scaffolds, which bind to the receptors identified below. Accordingly, the polypeptides of present invention may include commercially available antibodies or binding scaffolds, which have been designed to achieve specific binding to the target cell or receptor in question. Alternatively, preferred TMs include peptide ligands, such as cytokines, growth factors, neuropeptides, and lectins.

A TM of the present invention binds to a receptor on a cancer cell. By way of example, a TM of the present invention binds to a receptor on a cancer cell selected from the group comprising: a growth hormone-releasing hormone (GHRH, aka GHRF/GRF) receptor; an insulin-like growth factor receptor (e.g. an IGF-1 receptor); a corticotropin releasing factor receptor (e.g. CRHR-2); a gastrin-releasing peptide (GRP) receptor; a bombesin peptide (BB) receptor such as BRS-1, BRS-2, or BRS-3; a growth hormone (GH) receptor; an interleukin receptor (e.g. IL8RA or IL13RA1); a vascular endothelial growth factor (VEGF) receptor; an acetylcholine (ACH) receptor; a somatostatin (SST) receptor, such as SSTi, SST₂, SST₃, SST4 or SST₅; a cortistatin (CST) receptor; a chemokine (C-X-C motif) receptor such as CXCR4; a neuropilin receptor; a gonadotropin-releasing hormone (GnRH) receptor; a VIP-glucagon-GRF-secretin superfamily receptor, such as a PAC

-92015268754 16 Dec 2015 (eg. PACi) receptor or a vasoactive intestinal peptide VPAC receptor (e.g. VPAC-1 or VPAC-2) receptor; or an ErbB family member receptor such as an EGF receptor. All of these receptors are over-expressed in cancer cells.

In one embodiment of the present invention, the TM binds to a receptor on a cancer cell selected from the group comprising: FLT such as FLT1, FLT4, FLT3, BRS such as BRS3; CHRN such as CHRNA1, CHRND or CHRNE; EPHA such as EPHA7, EPHA4, EPHA5, EPHA3, EPHA1, EPHA2; EFN such as EFNB3, EFNA1, EFNB2, EFNA3, EFNB1; ErbB such as ERBB2, ERBB4;

DLK1, DLL3, FGF such as FGFR1, FGFR3, FGFR2; GRPR; GNRHR; GRPR; GnRHR; JAG such as JAG1 (CD339) or JAG2, IFGR such as IGF1R; leukaemia inhibitory factor receptor (LIFR); NMBR; NOTCHR such as NOTCH3 or NOTGH4; VIPR such as VIPR1; VEGFR such as VEGFR2; SSTR such as SSTR1; NMBR; or PDGFR such as PDGFRA or PDGFRB. All of these receptors are over-expressed in cancer cells.

In one embodiment, the TM is selected from: an adrenomedullin (ADM) peptide, an AM peptide, an autocrine motility factor (AMF) peptide, an amphiregulin peptide, an APRIL (a proliferation-inducing ligand) peptide, an artemin peptide, a betacellulin peptide, a bombesin peptide, a calcitonin receptor (CTR) binding peptide, an ErbB peptide such as an EGF peptide, an endothelin peptide, an erythropoietin peptide (EPO), a fibroblast growth factor (FGF) peptide such as a FGF-5 peptide, an FGF-18 peptide, a bFGF, a FGF8 or a FGF17 peptide, a follicle-stimulating hormone (FSH) peptide, a gastrin peptide, a gastrin releasing peptide (GRP), a glial cell line-defined neurotrophic factor (GDNF) peptide, a ghrelin (GHRL) peptide, a growth hormone-releasing hormone (GHRH) peptide, a granulocyte colonystimulating factor (G-CSF) peptide, a growth hormone (GH) peptide, a heparin-binding EGF-like growth factor (ΗΒ-EGF) peptide, a hepatocyte growth factor/ scatter factor (HGF/SF) peptide, an interleukin (IL) such as an IL-1 alpha peptide, an IL-6 peptide, an IL-8 peptide or an IL-10 peptide, an IGF-1 peptide, a stromal cell-derived factor-1 (SDF-1) peptide, a keratinocyte growth factor (KGF) peptide, a proepithelin/ granulin peptide, a leptin (LEP) peptide, a LIF peptide, an alpha-melanotropin peptide, a MGSA/GRO-alpha,

- 102015268754 16 Dec 2015 beta or gamma peptide, a NRG-1 alpha peptide, an oxytocin peptide, an osteopontin (OPN) peptide, a neuregulin-1 peptide, a PACAP peptide, a PDGF peptide such as a PDGF-alpha peptide, a PDGF-beta peptide, a PDGF-C peptide or a PDGF-D peptide, a prolactin peptide, a SCF peptide, a somatostatin peptide, a tumour necrosis factor (TNF) peptide such as a TNFalpha peptide, a TGF-beta peptide, a TGF-beta1 peptide, a VEGF peptide such as a VEGF-C peptide or a VEGF-D peptide, a vasopressin peptide, a VIP peptide, an angiopoietin peptide such as an angiopoietin-1 peptide or angiopoietin-2 peptide, a B-CLL peptide, a BCGF-12KD peptide, a BAFF peptide, a galanin peptide, a GDNF peptide, a GnRH peptide, an IGF-II peptide, a LH peptide, a neurotrophin peptide, a substance P peptide, or a TGF-alpha peptide; as well as truncations and peptide analogues thereof.

In one embodiment, a TM of the present invention binds to a leptin receptor.

Suitable examples of such TMs include: leptin peptides such as a full-length leptin peptide (eg. leptin^), and truncations or peptide analogues thereof such as Ieptin₂2-167, leptin₇o-95, and Ieptin₁₁₆-122In another embodiment, a TM of the present invention binds to a ghrelin receptor. Examples of suitable TMs in this regard include: ghrelin peptides such as full-length ghrelin (eg. ghrelin-117) and truncations or peptide analogues thereof such as ghrelin₂4-n7, and ghrelin₅2-n7; [Trp3, Arg5]-ghrelin (1-5), des-GIn-Ghrelin, cortistatin-8, His-D-Trp-Ala-Trp-D-Phe-Lys-NH₂, growth hormone releasing peptide (e.g. GHRP-6), or hexarelin.

In one embodiment, a TM of the present invention binds to a somatostatin (SST) receptor. By way of example, suitable TMs include: SST peptides and cortistatin (CST)-peptides, as well as peptide analogues thereof such as DPhe-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-NH₂ (BIM 23052), D-Phe-Phe-Tyr-D30 Trp-Lys-Val-Phe-D-Nal-NH₂ (BIM 23056) or c[Cys-Phe-Phe-D-Trp-Lys-ThrPhe-Cys]-NH₂ (BIM-23268). Further examples include the SST peptides SST14 and SST-28; as well as peptide and peptide analogues such as: octreotide, lanreotide, BIM23027, vapreotide, seglitide, and SOM230. These TMs are preferred TMs for binding to SST receptors, in particular to ssti, sst₂, sst₃, sst₄

- 11 2015268754 16 Dec 2015 and sst₅ receptors.

In another embodiment, a TM of the present invention binds to a gonadotropin-releasing hormone (GnRH) receptor. GnRH is also known as

Luteinizing-Hormone Releasing Hormone (LHRH). Examples of suitable GnRH receptor TMs include: GnRHI peptides, GnRHIl peptides and GnRHIII peptides, for example the full-length 92 amino acid GnRH precursor polypeptide and truncations thereof such as the decapeptide: pyroGlu-HisTrp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly CONH2.

In one embodiment, a TM of the present invention binds to insulin-like growth factor (IGF) receptor. Suitable examples include, for example IGF-1 peptides and IGF-2 peptides.

In one embodiment, a TM of the present invention binds to a VlP-glucagonGRF-secretin superfamily receptor, such as to a PAC (eg. PAC^ or to a VPAC (e.g. VPAC-1 or VPAC-2) receptor. Suitable examples of such TMs include pituitary adenylate cyclase-activating peptides (PACAP), vasoactive intestinal peptides (VIP), as well as truncations and peptide analogues thereof.

In one embodiment the TM is a VIP peptide including VIP-1 and VIP-2 peptides, for example VIP(1-28), or a truncation or peptide analogue thereof. These TMs demonstrate a selective binding to VPAC-1. Alternatively, a TM demonstrating a selective binding to VPAC2 may be employed, such as, for example mROM (see Yu et al., Peptides 27 (6) p1359-66 (2006), which is hereby incorporated by reference thereto). In another embodiment, the TM may be a PACAP peptide, for example PACAP(1-38) or PACAP(1-27), or a truncation of peptide analogue thereof. These TMs are preferred TMs for binding to PAC (eg. PAC-1) receptors.

In one embodiment, a TM of the present invention binds to an interleukin receptor. Suitable TM examples include: IL-1 peptides (e.g. IL-1 a, IL-β, IL-18 peptides) and truncations or peptide analogues thereof, IL-2 peptides (e.g. IL- 122015268754 16 Dec 2015

2, IL-3, IL-12, IL-23 peptides) and truncations or peptide analogues thereof, IL-6 and IL-8 peptides and truncations or peptide analogues thereof, and IL17 peptides (e.g. 11-17A, IL-17C peptides) and truncations or peptide analogues thereof.

In another embodiment, a TM of the present invention binds to an NGF receptor. Examples of suitable TMs include full-length NGF, and truncations or peptide analogues thereof.

In one embodiment, a TM of the present invention binds to a vasoactive epidermal growth factor (VEGF) receptor. Examples of suitable TMs include: VEGF peptide (e.g. VEGF-A, VEGF-B, VEGF-C, VEGF-D or VEGF-E and associated splice variants) and truncations or peptide analogues thereof, and placental growth factor (PIGF) and truncations or peptide analogues thereof.

In another embodiment, a TM of the present invention binds to an ErbB receptor. By way of example, the TM is selected from EGF peptides, transforming growth factor-α (TGF-a) peptides, chimeras of EGF and TGF-a, amphiregulin peptides, betacellulin peptides, epigen peptides, epiregulin peptides, heparin-binding EGF (ΗΒ-EGF) peptides, neuregulin (NRG) peptides such as NRG1a, NRG13, NRG2a, NRG23, NRG3, NRG4 and neuroregulin splice variants, tomoregulin 1 and 2 peptides, neuroglycan-C peptides, I in-3 peptides, vein peptides, gurken peptides, spitz peptides, or keren peptides; as well as truncations and peptide analogues thereof. There are 4 classes of ErbB receptor (termed ErbB1, erbB2, erbB3 and erbB4), which are also referred to as HER receptors. A number of variants of these receptors exist, which arise from alternate splicing and/ or cleavage of the fulllength receptor (eg EGFR v1 translation starts at aa543; EGFR vll deletion of aa521-603; EGFR vlll deletion of aa 6-273; EGFRvlll/A12-13 deletion of aa 630 273 and 409-520; EGFR vIV deletion of aa 959-1030; EGFR vV truncation at residue 958; EGFR TDM/2-7 tandem duplication of 6-273; EGFR TDM/18-25 tandem duplication of 664-1030; EGFR-TDM/18-26 tandem duplication of 664-1014). In addition, there are four ErbB4 receptor isoforms called erbB4

- 132015268754 16 Dec 2015

JM-a, erbB4 JM-b, erbB4 CYT-1 and erbB4 CYT-1.

Preferred TMs bind to ErbB receptors (eg. ErbB1, ErbB2, ErbB3, ErbB4) and splice variants thereof, in particular the ErbB1 receptor. ErbB TMs may also include proteins which contain EGF motifs with a splice site between the fourth and fifth cysteines within the six cysteine EGF-module, where this module is placed in close proximity to the transmembrane region of the potential ligand. For example, interphotoreceptor matrix proteoglycan-2 (IMP2), meprin (ΜΕΡ)1α, ΜΕΡ1β, mucin (MUC)3, MUC4, MUC12. and MUC17, as well as proteins with a T-knot scaffold such as potato carboxypeptidase inhibitor, and antibodies to ErbB receptors such as cetuximab, ABX-EGF, trastuzumab, or EMD72000. Further examples include chimeras generated by swapping domains (loop sequences and/or connecting amino acids) of different ErbB ligands, such as a mammalian erbB receptor ligand in which the B-loop sequence has replaced by those present in the insect (Drosophila) ErbB ligands, an ErbB ligand in which the C-loop sequence of EGF has been replaced by that of TGFa(44-50), EGF ligands in which one or more domain has been replaced by corresponding sequences in TGFa to create EGF/TGFa chimeras (e.g. E3T, T3E, E4T, T4E, T3E4T, T6E and E3T4E, and EGF chimeras in which the N-terminal TGFa sequence (WSHFND) or the neuregulin sequence (SHLVK) has been used to replace the N-terminal EGF sequence C-terminal of the first cysteine residue (NSDSE), T1E, and Biregulin. Yet further chimeras include EGF in which a domain has been replaced by an EGF-like domain of another protein, such as a blood coagulation, neural development or cell adhesion protein (e.g. Notch 3, Delta 1, EMR1, F4/80, EMR3 and EMR4 receptors).

In another embodiment, a TM of the present invention binds to a growthhormone-releasing hormone (GHRH) receptor. GHRH is also known as growth-hormone-releasing factor (GRF or GHRF) or somatocrinin. Suitable TM examples of the present invention include the full-length GHRH (1-44) peptide, and truncations or peptide analogues thereof such as GHRH(1-29), GHRH(1-37), hGHRH(1-40)-OH,

- 142015268754 16 Dec 2015 [MeTyrl, Ala 15,22,Nle27]-hGHRH(1 -29)-NH2 [MeTyrl ,Ala8,9,15,22,28,Nle 27j-hGHRH(1-29)-NH2 cyclo(25-29)[MeTyr1, Ala 15,DAsp25,Nle27,Orn29+++]-hGHRH(1 -29)-NH2. (D-Tyrl)-GHRH (1-29)-NH2 (D-Ala2)-GHRH (1-29)-NH2 (D-Asp3)-GHRH (1-29)-NH2 (D-Ala4)-GHRH (1-29)-NH2 (D-Thr7)-GHRH (1-29)-NH2 (D-Asn8)-GHRH (1-29)-NH2 (D-Ser9)-GHRH (1-29)-NH2 (D-TyrlO)-GHRH (1-29)-NH2 (Phe4)-GHRH (1-29)-NH2 (pCI-Phe6)-GHRH (1-29)-NH2 (N-Ac-Tyr1 )-GHRH (1-29)-NH2 (N-Ac-Tyr1, D-Ala2)-GHRH (1-29)-NH2 (N-Ac-D-Tyr1, D-Ala2)-GHRH (1-29)-NH2 (N-Ac-D-Tyr1, D-Ala 2, D-Asp3)-GHRH (1-29)-NH2 (D-Ala2, NLeu27)-GHRH (1-29)-NH2 (His1, D-Ala2, NLeu27)-GHRH (1-29)-NH2 (N-Ac-His1, D-Ala2, N-Leu27)-GHRH (1 -29)-NH2 (His1, D-Ala 2, D-Ala 4, Nleu27)-GHRH (1-29)-NH2 (D-Ala2, D-Asp3, D-Asn8, NLeu27)-GHRH (1-29)-NH2 (D-Asp3, D-Asn8, NLeu27)-GHRH (1-29)-NH2 [His1, NLeu27]-hGHRH(1-29)-NH2 [NLeu27J-hGHRH(1-29)-NH2

H-Tyr-Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-GIn-LeuSer-Ala-Arg-Lys-Leu-Leu-GIn-Asp-lle-Met-Ser-Arg-GIn-GIn-Gly-Glu-Ser-AsnGln-Glu-Arg-Gly-Ala-Arg-Ala-Arg-Leu-NH2

H-Tyr-Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-GIn-Leu30 Ser-Ala-Arg-Lys-Leu-Leu-Gln-Asp-lle-Met-Ser-Arg-NH2

H-Tyr-D-Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-GIn-LeuSer-Ala-Arg-Lys-Leu-Leu-Gln-Asp-lle-Met-Ser-Arg-NH2

H-Tyr-Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-lle-Leu-Gly-GIn-Leu-Ser

Ala-Arg-Lys-Leu-Leu-GIn-Asp-lle-Met-Asn-Arg-GIn-GIn-Gly-Glu-Arg-Asn-GIn152015268754 16 Dec 2015

Glu-Gln-Gly-Ala-Lys-Val-Arg-Leu-NH2

H-Tyr-Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-GIn-LeuSer-Ala-Arg-Lys-Leu-Leu-GIn-Asp-lle-Met-Asn-Arg-GIn-GIn-Gly-Glu-Arg-AsnGln-Glu-Gln-Gly-Ala-Lys-Val-Arg-Leu-NH2

His-Val-Asp-Ala-lie-Phe-Thr-GIn-Ser-Tyr-Arg-Lys-Val-Leu-Ala-GIn-Leu-SerAla-Arg-Lys-Leu-Leu-GIn-Asp-lle-Leu-Asn-Arg

His-Val-Asp-Ala-lle-Phe-Thr-GIn-Ser-Tyr-Arg-Lys-Val-Leu-Ala-GIn-Leu-SerAla-Arg-Lys-Leu-Leu-GIn-Asp-lle-Leu-Asn-Arg-GIn-GIn-Gly-Glu-Arg-Asn-GInGlu-GIn-Gly-Ala.

In a further embodiment, the TM binds to a bombesin receptor (eg. BRS-1, BRS-2, or BRS-3). TMs for use in the present invention that bind to a bombesin receptor include: bombesin - a 14 amino acid peptide originally isolated from the skin of a frog (pGlu-GIn-Arg-Leu-Giy-Asn-GIn-Trp-ASa-Val-Gly-His-Leu15 Met-NH₂); and the two known homologues in mammals, namely neuromedin B, and gastrin releasing peptide (GRP) such as porcine GRP - Aia-Pro-ValSer-Vai-GSy-Giy-GSy-Thr-Val-Leu-Ala-Lys-Mef-Tyr-Pro-Arg-Gly-Asn-His-TrpA!a-Vai-Giy-His-Leu-Met-NH2, and human GRP - Vai-Pro-Leu-Pro-Aia-GiyGly-Giy-Thr-Vai-Leu-Thr-Lys-Met-Tyr-Pro-Arg-GSy-Asn-HiS’Trp-Aia-Val-Gly20 His-Leu-Mel-NHs. Additional TMs include corresponding bombesin, neuromedin B and GRP truncations as well as peptide analogues thereof.

In one embodiment, the cancer cell is a lung cancer cell (e.g. a small cell lung cancer cell, a non-small cell lung cancer cell, or a carcinoid lung cancer cell).

The present inventors have confirmed that undesirable SNARE expression is observed in lung cancer cells. In this embodiment, the TM ligand binds to a receptor on the lung cancer cell, such as to a receptor selected from: an erythropoietin (EPO) receptor, a VEGF receptor such as a VEGF-3 receptor, an ErbB receptor such as an EGF receptor, an ErbB2 or 3 receptor, an GRP receptor, an ET(A) receptor, a CCK receptor such as a CCK-A or CCK-B receptor, a FLT receptor such a FLT 3 or FLT4 receptor, a CHRND receptor, an EPHA receptor such as an EPHA7, an EPHA4 or an EPHA5 receptor, an EFNA receptor such as an EFNA3 or EFNB3 receptor, a DLK1 receptor, an FGF receptor such as a FGF1 receptor, or a JAG receptor such as a JAG1

- 162015268754 16 Dec 2015 (CD339) or a JAG2 receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: erythropoietin (EPO), VEGF such as VEGF-B, VEGF-C, acetylcholine, ephrin peptides such as ephrin-A1,

A2, A3, A4 or A5, pro-neuregulin peptides such as pro-neuregulin-2, neuregulin peptides such as neuregulin or NTAK, EGF, TGF such as TGFbeta, GRP, bombesin like peptide, endothelin, PGF, TNF such as TNF-alpha, IL such as IL-6, IL-8, oxytocin, vasopressin, NRG such as NRG-1 alpha, bradykinin, HGF, GHRH, FGF such as bFGF, aFGF, FGF-1, FGF-2, NOTCH peptide ligands, FLT3 cytokine, or gastrin; as well as truncations and peptide analogues thereof.

In another embodiment the cancer cell is a bladder cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in bladder cancer cells. In this embodiment, the TM ligand binds to a receptor on the bladder cancer cell, such as to a receptor selected from: an IGF receptor such as an IGF1 receptor, a G-CSF receptor, a VEGF receptor such as a VEGF-2 receptor, an ErbB receptor such as an EGF receptor, a HGF receptor, or a FGF receptor such as a high/low affinity bFGF receptor.

Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: G-CSF, VEGF, ΗΒ-EGF, IGF such as IGF-1 or IGF-2, amphiregulin, betacellulin, hepatocyte growth factor (HGF), ErbB such as EGF, TGF such as TGF-apha, IL such as IL-6, granulin peptide such as granulin-4, or FGF such as bFGF; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a breast cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed In breast cancer cells. In this embodiment, the TM ligand binds to a receptor on the breast cancer cell, such as to a receptor selected from: an IGF receptor such as an IGF1 receptor, a VIP receptor such as VIPR-1, a GRP receptor, a NMB receptor, a CXC receptor such as CXCR4, a TNF receptor such as TNF receptor 1 or 2, a VEGF receptor such as VEGFR-2 or VEGFR-3, a neuropilin

- 172015268754 16 Dec 2015 receptor such as NRP-1 or NRP-2, an integrin receptor such as integrin receptor alpha9beta1, an OB receptor, an ET receptor such as ET-RA or ETRB, an erythropoietin receptor (EpoR), a TGF-beta receptor, an AMF receptor, or a prolactin receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: TNF such as TNF-alpha, VEGF such as VEGF- A, VEGF-B, VEGF-C, IL such as IL-6, amphiregulin, leptin, endothelin such as ET-1, ET-2, ET-3, FGF such as FGF-2, GHRH, granulin peptide such as granulin-4, erythropoietin, neuromedin peptides such as GRP neuromedin C, neuromedin B, autocrine motility factor (AMF), prolactin, growth hormone such as hGH, IGF such as IGF-1 or IGF-2, VIP peptides, or PACAP peptides such as PACAP-27 or PACAP-38; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a pancreatic cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in pancreatic adenocarcinoma cells. In this embodiment, the TM ligand binds to a receptor on the pancreatic cancer cell, such as to a receptor selected from: a VIP receptor such as a VIP1 receptor, a VEGF receptor such as a VEGF-1 or

VEGF2 receptor, a SST receptor such as a SST1 receptor, a CHRN receptor such as CHRNG, CHRNE, CHRNA1 or CHRND, an IGF receptor such as IGF1R, a BRS receptor such as BRS3, a GnRH receptor, a GRP receptor, a NMB receptor, a type-1 growth factor receptor, an ErbB receptor such as EGFR, a CCK receptor such as CCKB or CCKC, a PDGF receptor such as

PDGF-alpha, an ADM receptor, a keratinoctye growth factor (KGF) receptor such as FGFR2b, a type I FGF receptor, a GnRH receptor, a GFRalpha3/RET receptor, or a GDNF receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: ErbB such as EGF, TGF such as TGF-alpha, gastrin, PDGF, adrenomedullin, VEGF, KGF, FGF such as FGF-5 or FGF (a/b), VIP, SST peptides such as SST-14 or SST-28, acetylcholine, neuromedin peptides such as GRP neuromedin C, neuromedin B, PACAP peptided such as PACAP-27 or PACAP-38, IL such as IL-1 a, GnRH, bombesin, artemin, GDNF, or IGF such as IGF-1 or IGF-2; as well as

- 182015268754 16 Dec 2015 truncations and peptide analogues thereof.

In one embodiment the cancer cell is a prostate cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in prostate carcinoma cells. In this embodiment, the TM ligand binds to a receptor on the prostate cancer cell, such as to a receptor selected from: a CHRN such as a CHRNG or CHRNE receptor, an IGF receptor such as an IGF1 receptor, a VIP receptor such as a VIP1 receptor, a CT receptor, an ErbB receptor such as an EGFR (MR2), a [p75(NTR)] or TrkA receptor, a CC receptor such as a CCR2, a FGF receptor such as FGFR-1 to -4, a KGF receptor, a FSH receptor, a GHS receptor such as GHS-R 1a, a CXC receptor such as CXCR2, a VPAC or PAC receptor such as VPAC1 or PAC1, a CRL receptor, a c-Met/HGF receptor, or a GH receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: calcitonin (CT), TGF such as TGF-alpha, adrenomedullin, acetylcholine, prolactin, IL such as IL-6 or IL-8, NGF, PDF, MCP such as MCP-1, somatostatin, FGF such as FGF1 (acidic FGF), FGF2 (basic FGF), FGF6 or FGF8, EPO, VEGF, KGF, FSH, ghrelin, FGF17, TNF such as TNF-alpha, VIP,

PACAP peptides such as PACAP-27 or PACAP-38, AM, HGF, GH, GM-CSF, or IGF such as IGF-1 or IGF-2; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a cervix or uterine cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in cervix cancer cells. In this embodiment, the TM ligand binds to a receptor on the cervix or uterine cancer cell, such as to a receptor selected from: EPHA receptor such as EPHA5, EPHA7, EPHA4, EPHA3 receptor, a PDGF receptor such as PDGFRA, an EFNA receptor such as EFNA1, an SST receptor such as SSTR1, ab ErbB receptor such as an EGFR, a FLT receptor such as a FLT-1 receptor, a FLK receptor such as a FLK-1 receptor, a VEGF receptor such as VEGFR-3, an ET receptor such as ET(A)R, an IGF receptor such as IGF-1 R, or an LIF receptor such as LIFR. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as

- 192015268754 16 Dec 2015 well as truncations and peptide analogues thereof. Examples include; ErbB such as EGF, TGF such as TGF-alpha, G-CSF, PDGF peptides such as PDGFA or PDGFC, SST peptides such as SST-14 or SST-28, IGF such as IGF-1 or IGF-2, ephrin peptides such as ephrin-A1, A2, A3, A5 or A5, VEGF such as VEGF-C or VEGF-D, granulin peptide such as granulin-4, endothelin such as ET-1, or IL such as IL-6 or IL-1; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a leukaemia cancer cell. The present 10 inventors have confirmed that undesirable SNARE expression is observed in leukaemia cells. In this embodiment, the TM ligand binds to a receptor on the leukaemia cell, such as to a receptor selected from; an FLT receptor such as an FLT1 or 3 receptor, an FGF receptor such as FGFR1, 2 or 3, an EFN receptor such as an EFNB1 or EFNB3 receptor, a JAG receptor such as a

JAG1 (cd339) receptor, a PDGF receptor such as PDGFRA or B, EPHA1, EPHA2, a NOTCH receptor such as a NOTCH3 or 4 receptor, an EPHA receptor such as an EPHA7 receptor, a LIF receptor, an EFN receptor such as an EFNA1 or 3 receptor, a DLL receptor such as a DLL3 receptor, an ErbB receptor such as an ErbB2 or 3 receptor, a KDR receptor, a GHS receptor such as GHS-R type 1a, an IL receptor such as IL-1 receptor type II, or an IGF receptor such as an IGF1R. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: IGF such as IGF-1 or IGF-2, VEGF peptides such as VEGFB, ghrelin, APRIL, pro-neuregulin peptides such as pro-neuregulin-2, neuregulin peptides, NTAK, FGF such as bFGF, aFGF or FGF-1, NOTCH peptides such as NOTCH1, 2 or 3, PGF peptides, PDGF peptides such as PDGFA, PDGFB or PDGFD, ephrin peptides such as ephrin-A (1-5), LIF, GP30 ligand, erythropoietin, JAG peptides such as JAG-1 or JAG-2, Delta-1, IL such as IL1-alpha, or GM-CSF; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is an ovary cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in ovarian cancer cells. In this embodiment, the TM ligand binds to a receptor on

-202015268754 16 Dec 2015 the ovary cancer cell, such as to a receptor selected from: an ErbB receptor such as an ErbB2, 3 or 4 receptor, an FGF receptor such as FGFR3, a JAG receptor such as a JAG2 receptor, an EPH receptor such as an EPHA1 or 2 receptor, an IGF receptor such as an IGF1R, a GRP receptor, an EFN receptor such as an EFNB3, A1 or B2 receptor, or a GNRH receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: NRG peptides such as NRG-2 or NRG-3, or heparinbinding EGF-like growth factor peptides; neuregulin peptides, GP30 peptide,

NOTCH peptides, granulin peptides such as granulin-4, ephrin-A peptides suchas ephrin A1-A5, IGF peptides such as IGF-1 or fGF-2, NTAK, GRP, or GnRH; as well as truncations and peptide analogues thereof.

In one embodiment the cancer ceil is a bone cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in bone cancer cells. In this embodiment, the TM ligand binds to a receptor on the bone cancer cell, such as to a receptor selected from: an IGF receptor such as an IGF-IR, a GHRH receptor, or a FGF receptor such as a bFGF receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: PDGF such as PDGF-alpha or PDGF-beta, TGF such as TGFbetal, IGF such as IGF-1, GHRH, FGF such as bFGF, G-CSF, or IL such as IL-6; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a brain cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in brain cancer cells. In this embodiment, the TM ligand binds to a receptor on the brain cancer cell, such as to a receptor selected from: an EPO receptor, receptor 78 kDa glycoprotein (gp78), an ErbB receptor such as EGFR, as

PDGF receptor such as PDGFR-alpha, a CRL receptor such as CRLR/RAMP2 or CRLR/RAMP3, a neuropilin receptor such as NRP-1 or NRP-2, a CXC receptor such as CXCR4, a VEGFR such as VEGFR-1, an IGFR such as IGF1R, a GDNF receptor such as GDNFR-alpha 1, or a MET receptor. Preferred TMs in this embodiment include the corresponding natural

-21 2015268754 16 Dec 2015 ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: erythropoietin (EPO) peptides, IGF peptides, autocrine motility factor (AMF) peptides, GDNF peptides, HB-EGF peptides, TGF peptides such as TGF-alpha, PDGF peptides such as PDGFA, PDGF-B,

PDGF-C or PDGF-D, neuregulin peptides such as neuregulin-1, adrenomedullin, IL peptides such as IL-6, scatter factor/hepatocyte growth factor (SF/HGF) peptides, granuiin peptides such as granulin-4, VEGF peptides such as VEGF-A, or TGF peptides such as TGF beta 1 or TGF beta 2; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a colorectal cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in colorectal cancer cells In this embodiment, the TM ligand binds to a receptor on the colorectal cancer cell, such as to a receptor selected from: an IGF receptor such as IGF-R1, an ErbB receptor such as EGFR, a GRP receptor, an IL receptor such as IL6-R, a CCK receptor such as CCK-B receptor, or a prolactin receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: FGF such as FGF18, IGF such as IGF-1, TGF such as TGF-alpha, GRP, IL such as IL6, ErbB such as EGF, gastrin, or prolactin; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a liver cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in liver cancer cells In this embodiment, the TM ligand binds to a receptor on the liver cancer cell, such as to a receptor selected from: a TrkA (NGF) receptor, an IGF receptor such as IGF-IR, a HGF receptor such as a HGF-Met receptor, a c-met receptor, a gp78 receptor, or an ErbB receptor such as an EGFR. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: NGF, IL such as IL-6 or IL-8, IGF such as IGF-1 or IGF-2, HGF, SF/HGF, hepatopoietin (HPO), AMF, TGF such as TGF-beta or TGFalpha, LIF or PDGF; as well as truncations and peptide analogues thereof.

-222015268754 16 Dec 2015

In one embodiment the cancer cell is a skin cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in skin cancer cells In this embodiment, the TM ligand binds to a receptor on the skin (eg. basal, melanoma, squamous) cancer cell, such as to a receptor selected from: a FGF receptor such as FGFR-1, a c-kit receptor, a VEGF receptor such as VEGFR-2, a c-Met receptor, or a melanocortin receptor such as a melanocortin-1 receptor. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: IL such as IL8 or IL-10, FGF such as bFGF, SCF, VEGF such as VEGF-A, ErbB such as EGF, EPO, osteopontin (OPN), TGF such as TGF-beta, MGSA/GRO such as MGSA/GRO alpha, beta or gamma, HGF/SF, alpha-melanotropin, amphiregulin, or AMF; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a Kaposi sarcoma cancer cell. The present inventors have confirmed that undesirable SNARE expression is observed in Kaposi sarcoma cancer cells In this embodiment, the TM ligand binds to a receptor on the Kaposi sarcoma cell, such as to a receptor selected from: a PDGF receptor such as PDGFRA, a c-kit receptor, a TGF-beta receptor such as TGFR-1, an endothelin receptor such as ETA-R, a chemokine receptor such as CXCR3 or CCRL2, a VEGF receptor such as VEGFR-2, an IGF receptor such as IGF-IR, or an angiopoietin receptor such as TIE2. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof.

Examples include: PDGFa, IGF-1, Ang2, IL such as IL6, VEGF such as VEGF-A, endothelin-1, TGF such as TGF-beta, CXCL11, CCL8/14; as well as truncations and peptide analogues thereof.

In one embodiment the cancer cell is a renal cancer cell. In this embodiment, the TM ligand binds to a receptor on the renal cancer cell, such as to a receptor selected from: an IGF receptor such as IGF-IR, a VEGF receptor, a CXC receptor, or an ErbB receptor such as an EGFR. Preferred TMs in this embodiment include the corresponding natural ligand to said receptors, as well as truncations and peptide analogues thereof. Examples include: IL such

-2324

2015268754 19 Dec 2017 as IL-6 or IL-8, IGF such as IGF-1 or IGF-2, TGF such as TGF-beta or TGF-alpha, VEGF such as VEGF-A, chemokines such as CXCL12; as well as truncations and peptide analogues thereof.

The above embodiments describe particular cancers in which the present inventors have demonstrated undesirable SNARE expression (eg. up-regulated SNARE expression). The present invention is not, however, limited to said specific cancer types, and embraces all cancer types in which SNARE expression occurs.

The above-described undesirable SNARE expression (eg. up-regulation) in cancer cells may optionally be linked to an up-regulation of one or more specific receptor types present on the cancer cells in question. In this regard, the present inventors have identified particular receptor types (detailed above), which are up15 regulated in the same cells in which undesirable SNARE expression is observed (eg. up-regulated).

Comprises/comprising and grammatical variations thereof when used in this specification are to be taken to specify the presence of stated features, integers, steps or components or groups thereof, but do not preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.

Polypeptide preparation

The polypeptides of the present invention comprise 3 principal components: a 'bioactive' (ie. a non-cytotoxic protease); a TM; and a translocation domain. The general technology associated with the preparation of such fusion proteins is often referred to as re-targeted toxin technology. By way of exemplification, we refer to: W094/21300; WO96/33273; WO98/07864; WO00/10598; WO01/21213; W006/059093; WOOO/62814; WO00/04926; WO93/15766; WOOO/61192; and WO99/58571. All of these publications are herein incorporated by reference thereto.

24a

2015268754 19 Dec 2017

In more detail, the TM component of the present invention may be fused to either the protease component or the translocation component of the present invention. Said fusion is preferably by way of a covalent bond, for example either a direct covalent bond or via a spacer/ linker molecule. The protease component and the translocation component are preferably linked together via a covalent bond, for example either a direct covalent bond or via a spacer/ linker molecule. Suitable spacer/linked molecules are well known in the art, and typically

2015268754 16 Dec 2015 comprise an amino acid-based sequence of between 5 and 40, preferably between 10 and 30 amino acid residues in length.

In use, the polypeptides have a di-chain conformation, wherein the protease 5 component and the translocation component are linked together, preferably via a disulphide bond.

The polypeptides of the present invention may be prepared by conventional chemical conjugation techniques, which are well known to a skilled person. By way of example, reference is made to Hermanson, G.T. (1996), Bioconjugate techniques, Academic Press, and to Wong, S.S. (1991), Chemistry of protein conjugation and cross-linking, CRC Press, Nagy et al., PNAS 95 p1794-99 (1998). Further detailed methodologies for attaching synthetic TMs to a polypeptide of the present invention are provided in, for example, EP0257742.

The above-mentioned conjugation publications are herein incorporated by reference thereto.

Alternatively, the polypeptides may be prepared by recombinant preparation of a single polypeptide fusion protein (see, for example, WO98/07864). This technique is based on the in vivo bacterial mechanism by which native clostridial neurotoxin (i.e. holotoxin) is prepared, and results in a fusion protein having the following ‘simplified’ structural arrangement:

NH₂ - [protease component] - [translocation component] - [TM] - COOH

According to WO98/07864, the TM is placed towards the C-terminal end of the fusion protein. The fusion protein is then activated by treatment with a protease, which cleaves at a site between the protease component and the translocation component. A di-chain protein is thus produced, comprising the protease component as a single polypeptide chain covalently attached (via a disulphide bridge) to another single polypeptide chain containing the translocation component plus TM.

Alternatively, according to W006/059093, the TM component of the fusion -252015268754 16 Dec 2015 protein is located towards the middle of the linear fusion protein sequence, between the protease cleavage site and the translocation component. This ensures that the TM is attached to the translocation domain (ie. as occurs with native clostridial holotoxin), though in this case the two components are reversed in order vis-a-vis native holotoxin. Subsequent cleavage at the protease cleavage site exposes the N-terminal portion of the TM, and provides the di-chain polypeptide fusion protein.

The above-mentioned protease cleavage sequence(s) may be introduced (and/ or any inherent cleavage sequence removed) at the DNA level by conventional means, such as by site-directed mutagenesis. Screening to confirm the presence of cleavage sequences may be performed manually or with the assistance of computer software (e.g. the MapDraw program by DNASTAR, Inc.). Whilst any protease cleavage site may be employed (ie.

clostridial, or non-clostridial), the following are preferred:

Enterokinase Factor Xa

TEV(Tobacco Etch virus)

Thrombin

PreScission (DDDDKf) (IEGR)/IDGRf) (ENLYFQfG) (LVPRjGS) (LEVLFQjGP).

Additional protease cleavage sites include recognition sequences that are cleaved by a non-cytotoxic protease, for example by a clostridial neurotoxin.

These include the SNARE (eg. SNAP-25, syntaxin, VAMP) protein recognition sequences that are cleaved by non-cytotoxic proteases such as clostridial neurotoxins. Particular examples are provided in US2007/0166332, which is hereby incorporated in its entirety by reference thereto.

Also embraced by the term protease cleavage site is an intein, which is a selfcleaving sequence. The self-splicing reaction is controllable, for example by varying the concentration of reducing agent present. The above-mentioned ‘activation’ cleavage sites may also be employed as a ‘destructive’ cleavage

-262015268754 16 Dec 2015 site (discussed below) should one be incorporated into a polypeptide of the present invention.

In a preferred embodiment, the fusion protein of the present invention may 5 comprise one or more N-terminal and/ or C-terminal located purification tags.

Whilst any purification tag may be employed, the following are preferred:

His-tag (e.g. 6 * histidine), preferably as a C-terminal and/ or N-terminal tag MBP-tag (maltose binding protein), preferably as an N-terminal tag

GST-tag (glutathione-S-transferase), preferably as an N-terminal tag His-MBP-tag, preferably as an N-terminal tag GST-MBP-tag, preferably as an N-terminal tag Thioredoxin-tag, preferably as an N-terminal tag CBD-tag (Chitin Binding Domain), preferably as an N-terminal tag.

One or more peptide spacer/ linker molecules may be included in the fusion protein. For example, a peptide spacer may be employed between a purification tag and the rest of the fusion protein molecule.

Thus, a third aspect of the present invention provides a nucleic acid (e.g. DNA) sequence encoding a polypeptide as described above (i.e. the second aspect of the present invention).

Said nucleic acid may be included in the form of a vector, such as a plasmid, which may optionally include one or more of an origin of replication, a nucleic acid integration site, a promoter, a terminator, and a ribosome binding site.

The present invention also includes a method for expressing the abovedescribed nucleic acid sequence (i.e. the third aspect of the present invention) in a host cell, in particular in E. coli.

The present invention also includes a method for activating a polypeptide of the present invention, said method comprising contacting the polypeptide with a protease that cleaves the polypeptide at a recognition site (cleavage site)

-272015268754 16 Dec 2015 located between the non-cytotoxic protease component and the translocation component, thereby converting the polypeptide into a di-chain polypeptide wherein the non-cytotoxic protease and translocation components are joined together by a disulphide bond. In a preferred embodiment, the recognition site is not native to a naturally-occurring clostridial neurotoxin and/ or to a naturallyoccurring IgA protease.

The polypeptides of the present invention may be further modified to reduce or prevent unwanted side-effects associated with dispersal into non-targeted areas. According to this embodiment, the polypeptide comprises a destructive cleavage site. The destructive cleavage site is distinct from the ‘activation’ site (i.e. di-chain formation), and is cleavable by a second protease and not by the non-cytotoxic protease. Moreover, when so cleaved at the destructive cleavage site by the second protease, the polypeptide has reduced potency (e.g. reduced binding ability to the intended target cell, reduced translocation activity and/ or reduced non-cytotoxic protease activity). For completeness, any of the ‘destructive’ cleavage sites of the present invention may be separately employed as an ‘activation’ site in a polypeptide of the present invention.

Thus, according to this embodiment, the present invention provides a polypeptide that can be controllably inactivated and/ or destroyed at an off-site location.

In a preferred embodiment, the destructive cleavage site is recognised and cleaved by a second protease (i.e. a destructive protease) selected from a circulating protease (e.g. an extracellular protease, such as a serum protease or a protease of the blood clotting cascade), a tissue-associated protease (e.g. a matrix metalloprotease (MMP), such as an MMP of muscle), and an intracellular protease (preferably a protease that is absent from the target cell).

Thus, in use, should a polypeptide of the present invention become dispersed away from its intended target cell and/ or be taken up by a non-target cell, the polypeptide will become inactivated by cleavage of the destructive cleavage site (by the second protease).

-282015268754 16 Dec 2015

In one embodiment, the destructive cleavage site is recognised and cleaved by a second protease that is present within an off-site cell-type. In this embodiment, the off-site cell and the target cell are preferably different cell types. Alternatively (or in addition), the destructive cleavage site is recognised and cleaved by a second protease that is present at an off-site location (e.g. distal to the target cell). Accordingly, when destructive cleavage occurs extracellularly, the target cell and the off-site cell may be either the same or different cell-types. In this regard, the target cell and the off-site cell may each possess a receptor to which the same polypeptide of the invention binds.

The destructive cleavage site of the present invention provides for inactivation/ destruction of the polypeptide when the polypeptide is in or at an off-site location. In this regard, cleavage at the destructive cleavage site minimises the potency of the polypeptide (when compared with an identical polypeptide lacking the same destructive cleavage site, or possessing the same destructive site but in an uncleaved form). By way of example, reduced potency includes: reduced binding (to a mammalian cell receptor) and/ or reduced translocation (across the endosomal membrane of a mammalian cell in the direction of the cytosol), and/ or reduced SNARE protein cleavage.

When selecting destructive cleavage site(s) in the context of the present invention, it is preferred that the destructive cleavage site(s) are not substrates for any proteases that may be separately used for post25 translational modification of the polypeptide of the present invention as part of its manufacturing process. In this regard, the non-cytotoxic proteases of the present invention typically employ a protease activation event (via a separate ‘activation’ protease cleavage site, which is structurally distinct from the destructive cleavage site of the present invention). The purpose of the activation cleavage site is to cleave a peptide bond between the non-cytotoxic protease and the translocation or the binding components of the polypeptide of the present invention, thereby providing an ‘activated’ di-chain polypeptide wherein said two components are linked together via a di-sulfide bond.

-292015268754 16 Dec 2015

Thus, to help ensure that the destructive cleavage site(s) of the polypeptides of the present invention do not adversely affect the ‘activation’ cleavage site and subsequent di-sulfide bond formation, the former are preferably introduced into polypeptide of the present invention at a position of at least 20, at least 30, at least 40, at least 50, and more preferably at least 60, at least 70, at least 80 (contiguous) amino acid residues away from the ‘activation’ cleavage site.

The destructive cleavage site(s) and the activation cleavage site are 10 preferably exogenous (i.e. engineered/ artificial) with regard to the native components of the polypeptide. In other words, said cleavage sites are preferably not inherent to the corresponding native components of the polypeptide. By way of example, a protease or translocation component based on BoNT/A L-chain or H-chain (respectively) may be engineered according to the present invention to include a cleavage site. Said cleavage site would not, however, be present in the corresponding BoNT native L-chain or H-chain. Similarly, when the Targeting Moiety component of the polypeptide is engineered to include a protease cleavage site, said cleavage site would not be present in the corresponding native sequence of the corresponding Targeting Moiety.

In a preferred embodiment of the present invention, the destructive cleavage site(s) and the ‘activation’ cleavage site are not cleaved by the same protease. In one embodiment, the two cleavage sites differ from one another in that at least one, more preferably at least two, particularly preferably at least three, and most preferably at least four of the tolerated amino acids within the respective recognition sequences is/ are different.

By way of example, in the case of a polypeptide chimera containing a Factor

Xa ‘activation’ site between clostridial L-chain and Hn components, it is preferred to employ a destructive cleavage site that is a site other than a Factor Xa site, which may be inserted elsewhere in the L-chain and/ or Hn and/ or TM component(s). In this scenario, the polypeptide may be modified to accommodate an alternative ‘activation’ site between the L-chain and H_N

- 302015268754 16 Dec 2015 components (for example, an enterokinase cleavage site), in which case a separate Factor Xa cleavage site may be incorporated elsewhere into the polypeptide as the destructive cleavage site. Alternatively, the existing Factor Xa ‘activation’ site between the L-chain and H_N components may be retained, and an alternative cleavage site such as a thrombin cleavage site incorporated as the destructive cleavage site.

When identifying suitable sites within the primary sequence of any of the components of the present invention for inclusion of cleavage site(s), it is preferable to select a primary sequence that closely matches with the proposed cleavage site that is to be inserted. By doing so, minimal structural changes are introduced into the polypeptide. By way of example, cleavage sites typically comprise at least 3 contiguous amino acid residues. Thus, in a preferred embodiment, a cleavage site is selected that already possesses (in the correct position(s)) at least one, preferably at least two of the amino acid residues that are required in order to introduce the new cleavage site. By way of example, in one embodiment, the Caspase 3 cleavage site (DMQD) may be introduced. In this regard, a preferred insertion position is identified that already includes a primary sequence selected from, for example, Dxxx, xMxx, xxQx, xxxD, DMxx, DxQx, DxxD, xMQx, xMxD, xxQD, DMQx, xMQD, DxQD, and DMxD.

Similarly, it is preferred to introduce the cleavage sites into surface exposed regions. Within surface exposed regions, existing loop regions are preferred.

In a preferred embodiment of the present invention, the destructive cleavage site(s) are introduced at one or more of the following position(s), which are based on the primary amino acid sequence of BoNT/A. Whilst the insertion positions are identified (for convenience) by reference to BoNT/A, the primary amino acid sequences of alternative protease domains and/ or translocation domains may be readily aligned with said BoNT/A positions.

For the protease component, one or more of the following positions is preferred: 27-31, 56-63, 73-75, 78-81, 99-105, 120-124, 137-144, 161-165,

- 31 2015268754 16 Dec 2015

169-173, 187-194, 202-214, 237-241, 243-250, 300-304, 323-335, 375-382, 391-400, and 413-423. The above numbering preferably starts from the Nterminus of the protease component of the present invention.

In a preferred embodiment, the destructive cleavage site(s) are located at a position greater than 8 amino acid residues, preferably greater than 10 amino acid residues, more preferably greater than 25 amino acid residues, particularly preferably greater than 50 amino acid residues from the Nterminus of the protease component. Similarly, in a preferred embodiment, the destructive cleavage site(s) are located at a position greater than 20 amino acid residues, preferably greater than 30 amino acid residues, more preferably greater than 40 amino acid residues, particularly preferably greater than 50 amino acid residues from the C-terminus of the protease component.

For the translocation component, one or more of the following positions is preferred: 474-479, 483-495, 507-543, 557-567, 576-580, 618-631, 643-650, 669-677, 751-767, 823-834, 845-859. The above numbering preferably acknowledges a starting position of 449 for the N-terminus of the translocation domain component of the present invention, and an ending position of 871 for the C-terminus of the translocation domain component.

In a preferred embodiment, the destructive cleavage site(s) are located at a position greater than 10 amino acid residues, preferably greater than 25 amino acid residues, more preferably greater than 40 amino acid residues, particularly preferably greater than 50 amino acid residues from the Nterminus of the translocation component. Similarly, in a preferred embodiment, the destructive cleavage site(s) are located at a position greater than 10 amino acid residues, preferably greater than 25 amino acid residues, more preferably greater than 40 amino acid residues, particularly preferably greater than 50 amino acid residues from the C-terminus of the translocation component.

In a preferred embodiment, the destructive cleavage site(s) are located at a position greater than 10 amino acid residues, preferably greater than 25

-322015268754 16 Dec 2015 amino acid residues, more preferably greater than 40 amino acid residues, particularly preferably greater than 50 amino acid residues from the Nterminus of the TM component. Similarly, in a preferred embodiment, the destructive cleavage site(s) are located at a position greater than 10 amino acid residues, preferably greater than 25 amino acid residues, more preferably greater than 40 amino acid residues, particularly preferably greater than 50 amino acid residues from the C-terminus of the TM component.

The polypeptide of the present invention may include one or more (e.g. two, 10 three, four, five or more) destructive protease cleavage sites. Where more than one destructive cleavage site is included, each cleavage site may be the same or different. In this regard, use of more than one destructive cleavage site provides improved off-site inactivation. Similarly, use of two or more different destructive cleavage sites provides additional design flexibility.

The destructive cleavage site(s) may be engineered into any of the following component(s) of the polypeptide: the non-cytotoxic protease component; the translocation component; the Targeting Moiety; or the spacer peptide (if present). In this regard, the destructive cleavage site(s) are chosen to ensure minimal adverse effect on the potency of the polypeptide (for example by having minimal effect on the targeting/ binding regions and/ or translocation domain, and/ or on the non-cytotoxic protease domain) whilst ensuring that the polypeptide is labile away from its target site/ target cell.

Preferred destructive cleavage sites (plus the corresponding second proteases) are listed in the Table immediately below. The listed cleavage sites are purely illustrative and are not intended to be limiting to the present invention.

Second protease	Destructive cleavage recognition sequence	site	Tolerated recognition sequence variance P4-P3-P2-P1-T-P1’-P2’-P3’
		P4	P3	P2	P1	P1’	P2’	P 3’
Thrombin	LVPRVGS	A,F,G,I,	A,F,G	P	R	Not D	Not	-

-332015268754 16 Dec 2015

		L,T,V or M	,I,L,T, V,W or A			or E	D or E
Thrombin	GRVG			G	R	G
Factor Xa	IEGRT	A,F,G,I, L,T,V or M	D or E	G	R			-
ADAM 17	PLAQATVRSSS
Human airway trypsin-like protease (HAT)	SKGRTSLIGRV
ACE (peptidyl- dipeptidase A)						Not P	Not D or E	N /A
Elastase (leukocyte)	MEATVTY	M, R	E	A, H	V, T	V, T, H	Y	-
Furin	RXR/KRT	R	X	R or K	R
Granzyme	IEPDT	I	E	P	D			-
Caspase 1		F,W,Y, L		H, A,T	D	Not P, E.D. Q. K or R		-
Caspase 2	DVADT	D	V	A	D	Not P. E.D. Q. K or R		-
Caspase 3	DMQDT	D	M	Q	D	Not P, E.D. Q. K or R		-
Caspase 4	LEVDT	L	E	V	D	Not P, E.D. Q. K or R		-
Caspase 5		LorW	E	H	D			-

-342015268754 16 Dec 2015

Caspase 6		V	E	H or I	D	Not P. E.D. Q. K or R		-
Caspase 7	DEVDT	D	E	V	D	Not P, E.D. Q. K or R		-
Caspase 8		I or L	E	T	D	Not P, E.D. Q. K or R		-
Caspase 9	LEHDT	L	E	H	D		———	-
Caspase 10	IEHDT	I	E	H	D	———	-	-

Matrix metalloproteases (MMPs) are a preferred group of destructive proteases in the context of the present invention. Within this group, ADAM17 (EC 3.4.24.86, also known as TACE), is preferred and cleaves a variety of membrane-anchored, cell-surface proteins to shed the extracellular domains. Additional, preferred MMPs include adamalysins, serralysins, and astacins.

Another group of preferred destructive proteases is a mammalian blood protease, such as Thrombin, Coagulation Factor Vila, Coagulation Factor IXa, Coagulation Factor Xa, Coagulation Factor Xla, Coagulation Factor Xlla, Kallikrein, Protein C, and MBP-associated serine protease.

In one embodiment of the present invention, said destructive cleavage site comprises a recognition sequence having at least 3 or 4, preferably 5 or 6, more preferably 6 or 7, and particularly preferably at least 8 contiguous amino acid residues. In this regard, the longer (in terms of contiguous amino acid residues) the recognition sequence, the less likely non-specific cleavage of the destructive site will occur via an unintended second protease.

- 35 2015268754 16 Dec 2015

It is preferred that the destructive cleavage site of the present invention is introduced into the protease component and/ or the Targeting Moiety and/ or into the translocation component and/ or into the spacer peptide. Of these four components, the protease component is preferred. Accordingly, the polypeptide may be rapidly inactivated by direct destruction of the noncytotoxic protease and/ or binding and/ or translocation components.

Polypeptide delivery

In use, the present invention employs a pharmaceutical composition, comprising a polypeptide, together with at least one component selected from a pharmaceutically acceptable carrier, excipient, adjuvant, propellant and/ or salt.

The polypeptides of the present invention may be formulated for oral, parenteral, continuous infusion, implant, inhalation or topical application. Compositions suitable for injection may be in the form of solutions, suspensions or emulsions, or dry powders which are dissolved or suspended in a suitable vehicle prior to use.

Local delivery means may include an aerosol, or other spray (eg. a nebuliser). In this regard, an aerosol formulation of a polypeptide enables delivery to the lungs and/or other nasal and/or bronchial or airway passages.

The preferred route of administration is selected from: systemic, laparoscopic and/ or localised injection (e.g. injection directly into the tumour).

In the case of formulations for injection, it is optional to include a pharmaceutically active substance to assist retention at or reduce removal of the polypeptide from the site of administration. One example of such a pharmaceutically active substance is a vasoconstrictor such as adrenaline. Such a formulation confers the advantage of increasing the residence time of polypeptide following administration and thus increasing and/or enhancing its effect.

-362015268754 16 Dec 2015

The dosage ranges for administration of the polypeptides of the present invention are those to produce the desired therapeutic effect. It will be appreciated that the dosage range required depends on the precise nature of the polypeptide or composition, the route of administration, the nature of the formulation, the age of the patient, the nature, extent or severity of the patient’s condition, contraindications, if any, and the judgement of the attending physician. Variations in these dosage levels can be adjusted using standard empirical routines for optimisation.

Suitable daily dosages (per kg weight of patient) are in the range 0.0001-1 mg/kg, preferably 0.0001-0.5 mg/kg, more preferably 0.002-0.5 mg/kg, and particularly preferably 0.004-0.5 mg/kg. The unit dosage can vary from less that 1 microgram to 30mg, but typically will be in the region of 0.01 to 1 mg per dose, which may be administered daily or preferably less frequently, such as weekly or six monthly.

A particularly preferred dosing regimen is based on 2.5 ng of polypeptide as the 1X dose. In this regard, preferred dosages are in the range 1X-100X (i.e.

2.5-250 ng).

Fluid dosage forms are typically prepared utilising the polypeptide and a pyrogen-free sterile vehicle. The polypeptide, depending on the vehicle and concentration used, can be either dissolved or suspended in the vehicle. In preparing solutions the polypeptide can be dissolved in the vehicle, the solution being made isotonic if necessary by addition of sodium chloride and sterilised by filtration through a sterile filter using aseptic techniques before filling into suitable sterile vials or ampoules and sealing. Alternatively, if solution stability is adequate, the solution in its sealed containers may be sterilised by autoclaving. Advantageously additives such as buffering, solubilising, stabilising, preservative or bactericidal, suspending or emulsifying agents and or local anaesthetic agents may be dissolved in the vehicle.

Dry powders, which are dissolved or suspended in a suitable vehicle prior to use, may be prepared by filling pre-sterilised ingredients into a sterile

-372015268754 16 Dec 2015 container using aseptic technique in a sterile area. Alternatively the ingredients may be dissolved into suitable containers using aseptic technique in a sterile area. The product is then freeze dried and the containers are sealed aseptically.

Parenteral suspensions, suitable for intramuscular, subcutaneous or intradermal injection, are prepared in substantially the same manner, except that the sterile components are suspended in the sterile vehicle, instead of being dissolved and sterilisation cannot be accomplished by filtration. The components may be isolated in a sterile state or alternatively it may be sterilised after isolation, e.g. by gamma irradiation.

Advantageously, a suspending agent for example polyvinylpyrrolidone is included in the composition/s to facilitate uniform distribution of the components.

Administration in accordance with the present invention may take advantage of a variety of delivery technologies including microparticle encapsulation, viral delivery systems or high-pressure aerosol impingement.

Definitions Section

Targeting Moiety (TM) means any chemical structure that functionally interacts with a Binding Site to cause a physical association between the polypeptide of the invention and the surface of a target cell. In the context of the present invention, the target cell is a cancerous cell, for example one in which autocrine stimulation is driving proliferation, survival, metastatic potential or angiogenesis. The term TM embraces any molecule (ie. a naturally occurring molecule, or a chemically/physically modified variant thereof) that is capable of binding to a Binding Site on the target cell, which

Binding Site is capable of internalisation (eg. endosome formation) - also referred to as receptor-mediated endocytosis. The TM may possess an endosomal membrane translocation function, in which case separate TM and Translocation Domain components need not be present in an agent of the present invention. Throughout the preceding description, specific TMs have

- 382015268754 16 Dec 2015 been described. Reference to said TMs is merely exemplary, and the present invention embraces all variants and derivatives thereof, which retain the basic binding (i.e. targeting) ability of the exemplified TMs.

In one embodiment, the cancer cell target of the present invention may be a cancer cell that does not secrete growth hormone (ie. no significant or detectable growth hormone is secreted therefrom).

As mentioned previously, preferred TMs include antibodies (eg. antibody fragments) and binding scaffolds; especially commercially avilable antibodies/ fragments and scaffolds designed for the purpose of binding (eg. specifically) to cancer cells.

Protein scaffolds represent a new generation of universal binding frameworks to complement the expanding repertoire of therapeutic monoclonal antibodies and derivatives such as scFvs, Fab molecules, dAbs (single-domain antibodies), diabodies and minibodies, each of which may be employed as a TM of the present invention. Scaffold systems create or modify known protein recognition domains either through creation of novel scaffolds or modification of known protein binding domains. Such scaffolds include but are not limited to:

(i) protein A based scaffolds - affibodies (Nord, K. et al 1997 “Binding proteins selected from combinatorial libraries of an alpha-helical bacterial receptor domain”. Nat Biotechnol 15, 772-777);

(ii) lipocalin based scaffolds - anticalins (Skerra 2008 “Alternative binding proteins: anticalins - harnessing the structural plasticity of the lipocalin ligand pocket to engineer novel binding activities”. FEBS J. 275:2677-83);

(iii) fibronectin based scaffolds - adnectin (Dineen et al 2008 “The Adnectin CT-322 is a novel VEGF receptor 2 inhibitor that decreases tumor burden in an orthotopic mouse model of pancreatic cancer”. BMC Cancer 8:352);

(iv) avimers (Silverman et al 2005 “Multivalent avimer proteins evolved by exon shuffling of a family of human receptor domains”. Nat Biotechnol 23:1556-61);

(v) ankyrin based scaffolds - darpins (Zahnd et al 2006 “Selection and

-392015268754 16 Dec 2015 characterization of Her2 binding-designed ankyrin repeat proteins”. J Biol Chem. 281:35167-75); and (vi) centyrin scaffolds - based on a protein fold that has significant structural homology to Ig domains with loops that are analogous to CDRs. Ig domains are a common module in human proteins and have been widely applied as alternative scaffold proteins. Each of the above ‘scaffold’ publications is hereby incorporated (in its entirety) by reference thereto.

Binding scaffolds can be used to target particular cell types via interaction with 10 specific cell surface proteins, receptors or other ceil surface epitopes such as sugar groups. Such modified scaffolds can be engineered onto recombinant non-cytotoxic protease based polypeptides of the present invention to target specific cancer cell types of interest.

The TM of the present invention binds (preferably specifically binds) to the target cell in question. The term “specifically binds” preferably means that a given TM binds to the target cell with a binding affinity (Ka) of 10⁶ M¹ or greater, preferably 10⁷ M‘¹ or greater, more preferably 10^θ M'¹ or greater, and most preferably, 10⁹ M'¹ or greater.

Reference to TM in the present specification embraces fragments and variants thereof, which retain the ability to bind to the target cell in question. By way of example, a variant may have at least 80%, preferably at least 90%, more preferably at least 95%, and most preferably at least 97 or at least 99% amino acid sequence homology with the reference TM. Thus, a variant may include one or more analogues of an amino acid (e.g. an unnatural amino acid), or a substituted linkage. Also, by way of example, the term fragment, when used in relation to a TM, means a peptide having at least ten, preferably at least twenty, more preferably at least thirty, and most preferably at least forty amino acid residues of the reference TM. The term fragment also relates to the above-mentioned variants. Thus, by way of example, a fragment of the present invention may comprise a peptide sequence having at least 10, 20, 30 or 40 amino acids, wherein the peptide sequence has at least 80% sequence homology over a corresponding peptide sequence (of contiguous) amino

-402015268754 16 Dec 2015 acids of the reference peptide.

By way of example, ErbB peptide TMs may be modified to generate mutein ErbB ligands with improved properties such as increased stability. By way of example, ErbB TM muteins include ErbB peptides having amino acid modifications such as a valine residue at position 46 or 47 (EGFVal46 or 47), which confers stability to cellular proteases. ErbB TMs may also have amino acids deleted or additional amino acids inserted. This includes but is not limited to EGF having a deletion of the two C-terminal amino acids and a neutral amino acid substitution at position 51 (particularly EGF51Gln51; see US20020098178A1), and EGF with amino acids deleted (e.g. rEGF2-48; rEGF3-48 and rEGF4-48). Fragments of ErbB TMs may include fragments of TGFa which contain predicted β-turn regions (e.g. a peptide of the sequence Ac-C-H-S-G-Y-V-G-A-R-C-O-OMe), fragments of EGF such as [Ala20]EGF(14-31), and the peptide YHWYGYTPQNVI or GE11. All of the above patent specifications are incorporated herein by reference thereto.

By way of further example, somatostatin (SST) and cortistatin (CST) have high structural homology, and bind to all known SST receptors. Full-length

SST has the amino acid sequence:

MLSCRLQCALAALSIVLALGCVTGAPSDPRLRQFLQKSLAAAAGKQELAKYF

LAELLSEPNQTENDALEPEDLSQAAEQDEMRLELQRSANSNPAMAPRERKA

GCKNFFWKTFTSC

Full-length CST has the amino acid sequence:

MYRHKNSWRLGLKYPPSSKEETQVPKTLISGLPGRKSSSRVGEKLQSAHKM

PLSPGLLLLLLSGATATAALPLEGGPTGRDSEHMQEAAGIRKSSLLTFLAWW

FEWTSQASAGPLIGEEAREVARRQEGAPPQQSARRDRMPCRNFFWKTFSS

CK

Reference to these TMs includes the following fragments (and corresponding variants) thereof:

NFFWKTF;

(R or K)NFFWKTF;

-41 2015268754 16 Dec 2015

C(R or K)NFFWKTF;

(P or G)C(R or K)NFFWKTF;

NFFWKTF(S orT);

NFFWKTF(S or T)S;

NFFWKTF(S or T)SC;

(Ror K)NFFWKTF(S or T);

(R or K)NFFWKTF(S or T)S;

(R or K)NFFWKTF(S or T)SC;

C(Ror K)NFFWKTF(S orT);

C(R or K)NFFWKTF(S or T)S;

C(R or K)NFFWKTF(S or T)SC;

(P or G)C(R or K)NFFWKTF(S or T);

(P or G)C(R or K)NFFWKTF(S or T)S; or (P or G)C(R or K)NFFWKTF(S or T)C.

With regard to the above sequences, where a (P or G) alternative is given, a P is preferred in the case of a CST TM, whereas a G is preferred in the case of an SST TM. Where an (R or K) alternative is given, an R is preferred in the case of a CST TM, whereas a K is preferred in the case of an SST TM. Where an (S or T) alternative is given, an S is preferred in the case of a CST TM, whereas a T is preferred in the case of an SST TM.

Preferred fragments comprise at least 7 or at least 10 amino acid residues, preferably at least 14 or at least 17 amino acid residues, and more preferably at least 28 or 29 amino acid residues. By way of example, preferred sequences include:

SANSNPAMAPRERKAGCKNFFWKTFTSC (SST-28);

AGCKNFFWKTFTSC (SST-14);

QEGAPPQQSARRDRMPCRNFFWKTFSSCK (CST-29); QERPPLQQPPHRDKKPCKNFFWKTFSSCK (CST-29); QERPPPQQPPHLDKKPCKNFFWKTFSSCK (CST-29)

DRMPCRNFFWKTFSSCK (CST-17);

PCRNFFWKTFSSCK (CST-14); and

- 42 2015268754 16 Dec 2015

PCKNFFWKTFSSCK (CST-14)

The TM may comprise a longer amino acid sequence, for example, at least 30 or 35 amino acid residues, or at least 40 or 45 amino acid residues, so long as the TM is able to bind to a normal GH-secreting cell, preferably to an SST or to a CST receptor on a normal GH-secreting cell. In this regard, the TM is preferably a fragment of full-length SST or CST, though including at least the core sequence ’’NFFWKTF” or one of the above-defined primary amino acid sequences.

It is routine to confirm that a TM binds to the selected target cell. For example, a simple radioactive displacement experiment may be employed in which tissue or cells representative of a growth hormone-secreting cell are exposed to labelled (eg. tritiated) TM in the presence of an excess of unlabelled TM. In such an experiment, the relative proportions of non-specific and specific binding may be assessed, thereby allowing confirmation that the TM binds to the target cell. Optionally, the assay may include one or more binding antagonists, and the assay may further comprise observing a loss of TM binding. Examples of this type of experiment can be found in Hulme, E.C.

(1990), Receptor-binding studies, a brief outline, pp. 303-311, In Receptor biochemistry, A Practical Approach, Ed. E.C. Hulme, Oxford University Press.

In the context of the present invention, reference to a peptide TM (e.g. GHRH peptide, or leptin peptide) embraces peptide analogues thereof, so long as the analogue binds to the same receptor as the corresponding ‘reference’ TM. Said analogues may include synthetic residues such as:

β-Nai - β-naphthylaianine β-Pa! ~ β -pyridyialanine hArg(Bu) ~ N-guanidino-(butyi)-homoarginine hArg(Et)z - N, N'-guanidino-(dimethyl)-homoarginine hArg(CH2CFs)2 ~ N, N-guanidino-bis-(2,2,2,-trifluoroethyl)-homoarginine hArg(GHs, hexyl) = N, N-guan!dino-(methyl, hexyl)- homoarginine Lys(Me) --- N^e-methyllysine

-432015268754 16 Dec 2015

Lys(iPr) ··' N®-isopropyllysine AmPhe ·- aminomethylphenyiaianine AChxAla ~ aminocyclohexyialanine Abu ~ α-aminobutyric acid

Tpo = 4-thiaproiine

MeLeu ~ N-methyileucine Orn = ornithine Nle - norleucine Nva ~ norvaiine

Trp(Br) ~ 5-bromo-tryplophan Trp(F) ~ 5-fluoro-tryptophan Trp(NOs) = 5-nitro-tryptophan Gaba ~ y-aminobutyric acid Bmp = J-mercaptopropionyl

Ac ~ acetyl

Pen - pencillamine

By way of example, the above peptide analogue aspect is described in more detail with reference ίο specific peptide TMs, such as SST peptides, GHRH peptides, bombesin peptides, GnRH peptides, and ghrelin peptides, though the same principle applies to ail TMs of the present invention..

Somatostatin analogues, which can be used to practice the present invention include, but are not limited to, those described in the following publications, which are hereby incorporated by reference: Van Binst, G. et al. Peptide Research 5: 8 (1992); Horvath, A. et al. Abstract, Conformations of Somatostatin Analogs Having Antitumor Activity, 22nd European peptide Symposium, September 13-19,1992, Interlaken, Switzerland; US5,506,339; EP0363589; U54,904,642; 034,871,717; US4,725,577; US4,684,620;

US4,650,787; US4,585,755; US4,725,577; 1184,522,813; US4,369,179;

US4,360,516: US4,328,214; US4,316,890; US4,319,518; US4,291,022;

US4,238,481: US4,235,888; 054,211,693,- US4,190,648; US4,146,612;

1184,133,782: USS,506,339; US4,261,885; US4,282,143; US4,190,575;

USS,552,520; EP0389180; EP0505680; US4,603,120; EP0030920;

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US4,853,371; WO90/12811; WG97/G1579; WO91/18016; WO98/08529 and WO98/08528: WO/0075186 and WOOO/G6185; WO99/56769; and FR 2,522,655. Each of these publications is incorporated in its entirety by reference thereto.

Methods for synthesizing analogues are well documented, as illustrated, for example, by the patents cited above. For example, synthesis of H-D-Phe-PhePhe-D-Trp-Lys-Thr~Phe-Thr-NH2, can be achieved by following the protocol set forth in Example 1 of EP0395417A1. Similarly, synthesis analogues with a substituted N-terminus can be achieved, for example, by following the protocol set forth in WO88/02756, EP0329295, and USS,240,561.

The use of linear SST analogues are also included within the scope of this invention, for example: H-D-Phe-p-chloro-Phe-Tyr-D-Trp-Lys-Thr-Phe-Thr15 NH2; H-D-Phe-p-N02-Phe-Tyr-D-Trp-Lys-Vai-Phe-Thr~NH2; H-D-*Na!~pch!oro-Phe-Tyr-D-Trp-Lys-Vai-Phe-Thr-NH2; H-D~Phe-Phe-Phe-D-Trp-LysThr-Phe-Thr-NH2; H-D-Phe-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH2; H-D-Phe-pchloro-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH2; and H-D-Phe-Ala-Tyr-D-Trp-LysVal-Ala-D-beta-Nal-NH2.

One or more chemical moieties, eg. a sugar derivative, mono or poly-hydroxy (C2-12) alkyl, mono or poly-hydroxy (C2-12) acyl groups, or a piperazine derivative, can be attached to a SST analogue, e g. to the N-terminus amino acid - see WO88/Q2756, EP0329295, and USS,240,561.

GHRH peptide analogues date back to the 1990s, and include the ‘standard antogonist’ [Ac-Tyr', D-Arg2jhGH-RH (1-29) Nha. US Patent 4, 659,693 discloses GH-RH antagonistic analogs which contain certain N, Ν'- dialkylomega-guanidino alpha-amino acyl residues in position 2 of the GH-RH (1-29) sequence. Additional examples are provided in WO91/16923, US 5,550,212, US5,942,489, US6,057,422 USS, 942,489, US6,057, 422, WO96/032126, WO967022782, W096/016707, WO94/011397, WO94/011396, each of which is herein incorporated by reference thereto.

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Examples of bombesin analogues suitable for use in the present invention include TMs comprising: D-Phe-GIn-Trp-Ala-Vai-Giy-His-Leu-Met-NHz (code named BiM-26218), D-Phe-GIn-Trp-Aia-Val-Giy-Hss-Leu-Leu-NHg (code named BiM-26187); D-Cpa-Gin-Trp-Aia-Vai-Giy-His-Leu-φ [CH₂NH]-Phe-NH₂ (code named BIM-26159), and D-Phe-Gin-Trp-Aia-Vai-Giy-His-Leu-φ [CH₂NH]-Cpa-NH₂ (code named BIM-26189); D-Phe-GIn-Trp-Ala-Val-Nmethyi-D-Aia-His-Leu- methyiester, and D-F_g-Phe-G!n--Trp-A!a-Va!-D-Ala-HisLeu- methyiester.

Bombesin analogues include peptides derived from the naturally-occurring, structurally-related peptides, namely, bombesin, neuromedin B, neuromedin C, litorin, and GRP. The relevant amino acid sequences of these naturally occurring TM peptides are listed below:

Bombesin (last 10 aa’s): Gly-Asn-Gln~Trp-Ala-Val-G!y-His-Leu~Met-NH₂ Neuromedin 8: Giy-Asn-Leu-Trp-Aia-Thr-Giy-His-Phe-Met-NH?

Neuromedin C: Gly-Asn~His-Trp-Ala-Vai-Giy~Hss~Leu-Met-NH₂

Litorin: pGlu-Gln-Trp-Ala-Val-Gly-His-Phe-Met-NH2

Human GRP (last 10 aa’s): Giy-Asn-His-Trp-Aia-Vai-Gly-His-Leu-Met-NH2

Analogs suitable for use in the present invention are described in U.S. Serial Number 502,438, filed March 30, 1990, U.S. Serial No. 397,169. filed August 21, 1989, U.S. Serial No. 376,555, filed July 7, 1989, U.S. Serial Number 394,727, filed August 16, 1989, U.S. Serial No. 317,941, filed March 2, 1989,

U.S. Serial Number 282,328, filed December 9, 1988, U.S. Serial No. 257,998, filed October 14, 1988, U.S. Serial No. 248,771, filed September 23, 1988, U.S. Serial No. 2.07,759, filed June 16, 1988, U.S. Serial No. 204,171, filed June 8, 1988, U.S. Serial No. 173,311, filed March 25, 1988, U.S. Serial No. 100,571, filed September 24, 1987; and U.S. Serial No. 520,225, filed

May 9, 1990, U.S, Serial No. 440,039, filed November 21, 1989. Ail these applications are hereby incorporated by reference. Bombesin analogs are also described in Zachary et al., Proc. Nat. Aca. Sci. 82:7616 (1985): Heimbrook et al., ’’Synthetic Peptides: Approaches to Bioiogicai Problems, UCLA Symposium on Mol. and Cell. Biol. New Series, Voi. 86, ed. Tarn and Kaiser;

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Heinz-Erian et ai., Am. J. Physiol. G439 (1986); Martinez et al., J. Med. Ghent. 28:1874 (1985); Gargosky et al., Biochem. J. 247:427 (1987): Dubreuil et ai. , Drug Design and Delivery, Vo! 2:49, Harwood Academic Publishers, GB (1987); Heikkiia et a!., J. Biol. Chem. 262:16456 (1987); Caranikas et al. , J.

Med. Ghent. 25:1313 (1982); Saeed et ai., Peptides 10:597 (1989); Resell et ai., Trends in Pharmacological Sciences 3:211 (1982); Lundberg et al., Proc. Nat, Aca. Sci. 80:1120, (1983); Engberg et al., Nature 293:222 (1984); Mizrahi et al., Euro. J. Pharma. 82:101 (1982); Leander et al., Nature 294:467 (1981); Woil et a!., Biochem. Biophys. Res. Comm. 155:359 (1988); Rivier et al.,

Biochem. 17:1766 (1978); Cuttitta et a!., Cancer Surveys 4:707 (1985); Aumelas et ah, Int. J. Peptide Res. 30:596 (1987): ail of which are also hereby incorporated by reference.

The analogs can be prepared by conventional techniques, such as those described in WO92/20363 and EP0737691. Additional bombesin analogues are described in, for example, WO89/02897, WO91/17181, W090/03980 and WO91 /02746, all of which are herein incorporated by reference thereto.

Examples of ghrelin analogues suitable for use as a TM of the present invention comprise: Tyr-DTrp-DLys-Trp-DPhe-NH₂, Tyr-DTrp-Lys-Trp-DPheNH₂, His-DTrp-DLys-Trp-DPhe-NHs, His-DTrp-DLys-Phe-DTrp-NH₂, HisDTrp-DArg-Trp-DPhe-NH₂, His~DTrp~DLys-Trp-DPhe-Lys~NH₂, DesaminoTyrDTrp-Ala-Trp-DPhe-NH_2> DesaminoTyr-DTrp-DLys-Trp-DPhe-NH₂,

DeaminoTyr-DTrp-Ser-Trp-DPhe-LyS’NHz, DesarninoTyr-DTrp-Ser-Trp-DPhe25 NH₂, His-DTrp-DTrp-Phe-Met-NHs, Tyr-DTrp-DTrp~Phe-Phe-NH₂, Giyqj[CH₂NH]-DpNal-Ala-Trp-DPheLys-NH₂, Glyip[CH2NH]-DbetaNal-DLyS~ TrP-DPhe-Lys-NH₂, DA!a-DbetaNal-DLys-DTrp~Phe-L.ys~NH₂. His-DbetaNalDLys-Trp-DPhe-Lys-NH₂, A!a-His-DTrp-DLys~Trp-DPhe-Lys-NH₂₎

Aia9[CH₂NH]-DbetaMal-A!a-Trp-DPhe-Lys-NH₂₍ DbeiaNal-Ala-Trp-DPhe-Ala30 NH₂, DAIa-DcyclohexylAla-Ala-Phe-DPhe~N!e-NH₂, DcyclohexylAla-Aia-PheDTrp-Lys-NH;?, DAia-DbetaASa-Thr-DThr-Lys-NH₂, DcyclohexylAia-Ala-TrpDPhe-NH2, DAIa-DbetaNal-A!a-Ala-DAIa-Lys-NH₂, DbetaNal-Ala-Trp-DPheLeu-NH₂, His-DTrp~Phe-Trp-DPhe-LyS’NH₂, DAIa-DbetaNai-DAIa-DTrp-PheLys-NH₂, pAla-Trp-DA!a-DTrp-Phe-NH₂, His-Trp~DAIa-DTrp-Phe-LysNH₂,

-472015268754 16 Dec 2015

DLys-DpNal-Aia-Trp-DPhe-Lys-NHz, DAla-DbetaNal-DLys-DTrp-Phe-LysNH₂, Tyr-DAIa-Phe-Aib-NHs, Tyr-DAia-Sar-NMePhe-NH₂, ayAbu-DTrp-DTrpSer-NHs, ayAbu-DTrp-DTrp-Lys-NH₂, ayAbu-DTrp-DTrp-Orn-MH₂, aAbuDTrp-DTrp-Orn-NH₂, DThr~D0Nal-DTrp-DPro-Arg~NH₂, DASa-Aia-DAIa-DTrp5 Phe-Lys-NH₂, A^[CH₂NH3Hi5-DTrp-ASa-Trp-DPhe-Lys-NH₂, Lys-DHis-DTrpPhe-NH?:, yAbu-DTrp-DTrp-Orn-NH₂, inip-Trp~Trp~Phe-NH₂, Ac-DTrp-PbeDTrp-Leu-NHi’, Ac-DTrp-Phe-DTrp-Lys-NH₂, Ac-DTrp-DTrp-Lys-NH₂, DLysTyr-DTrp-DTrp-Phe-Lys-NH₂, Ac-DbetaNai-Leu-Pro-NH₂, pAla-Trp-DTrpDTrp-Orn-NH₂₎ DVal-DaNal-DTrp-Phe-Arg-NHz, DLeu-DaNal-DTrp-Phe-Arg10 NH₂, CyciohexylAla-DaNai-DTrp- Phe-Arg-NH₂₎ DTp-DaNal-DTrp-Phe-ArgNH₂, DAia-DpNal-DPro-Phe-Arg-NHs, Ac-DaNa!-DTrp-Phe-Arg-NH₂, DaNaiDTrp-Phe-Arg-NH₂, His-DTrp-DTrp-Lys-NH₂, Ac-DpNal~DTrp-NH₂, aAib-DTrpDcyclohexyiAia~NH₂₎ aAib-DTrp-DAia-cyclohexy!Ala-NH₂₎ DAlaDcyclohexySAIa“Ala-AS3“Phe-DPhe-Nie-NH_2j DPhe-Aia-Phe-DPal-NH^ DPhe15 Ala-Phe-DPhe-Lys-NHs, DLys-Tyr-DTrp-DTrp-Phe-NH₂, Ac-DLys-Tyr-DTrpDTrp-Phe-NH₂, Arg-DTrp-Leu-Tyr-Trp-Pro(cyclic Arg-Pro), Ac-DpNaS-PtcLysILys-DPhe-NH2, DPahPhe-DTrp-Pbe-Met-NH₂, DPhe-Trp-DPhe-Phe-MetNH₂, DPal-Trp-DPhe-Phe-Met-NHs, pAia-Pa!-DTrp-DTrp-Orn-NH₂, ayAbiiTrp-DTrp-DTrp-Om-NH₂₎ pAla-Trp-DTrp-DTrp-Lys-NHg, yAbu-Trp-DTrp-DTrp20 Orn-NH₂₍ Ava-Trp-DTrp-DTrp-Orn-NH₂, DLys-Tyr-DTrp-Aia-Trp-DPhe-NH₂, His-DTrp-DArg-Trp-DPhe-NH₂, <G!u-His-Trp-DSer-DArg-NH₂, DPhe-DPheDTrp-Met-DLys-NHs, G-(2-methylaNyl) benzophenone oxime, (R)~2-amino-3(!H-indo!“3-yl)“l-(4“phenylpiperidin-1 -yi)propan-1 -one, N-((R)-1-((R)-1-((S)-3“ (SH4ndol-3-yl)-1 -oxo-1 -(4-pheny ipiperidin-1-y!)propan-2~yiamino)-6-am!no-125 oxohexan-2-y!amino)-3-hydroxy-1 -oxopropan-2-yi)benzamide, (S)-N-((S)-3(1 H-indo!-3-yi)-1 -oxo-1 -(4- phenyl piperid in - 4 -yi)propan-2-yi)-6-acetarnido-2((S)-2-amino-3-(benzyloxy)propanamido)hexanamide, (S)-N-((R)-3-(1 H-indol3-yi)-1-oxo-1-(4- phenySp!peridin-1~yi)propan-2-yl}-2-((S)~2-acetamido-3(benzyioxy)propanamido)-6-aminohexanamide, (R)-N-(3~(1 H-indoi-3~yl)-1 -(430 (2-methoxyphenyi)pipendin”1 -yi)-1 -oxopropan-2-yl)-4-amlrsobutanamide, (R)N-(3-(1 H-i ndol-3-yl)-1 ~(4~(2-methoxyphenyl)pipendin-1 -yi )-1 -oxopropan-2-yl)2-arnino-2-methyipropanamide, methyl 3-(p-tolylcarbamoyi)-2-naphthoate, ethyl 3-{4-(2-methoxyphenyl)piperidine-1 -carbonyi)-2-naphthoate, 3-(2methoxyphenyicarbamoyi)-2-naphthoate, (S)-2,4~diamino-N-((R)-3-482015268754 16 Dec 2015 (naphth3ien-2-ylmethoxy)-1-oxo~1-(4-phenylpiper!din-1-y!)propan-2yQbutanamide, naphthalene-2,3-diyibis((4-(2-methoxyphenyl)piperazin-1yi)methanone), (R)-2-amino-N-(3-(benzyloxy)-1-oxo-1-(4-phenylpiperazin-1yl)propan-2-yi)-2-methyipropanamide, or (R)-2-amino-3-(benzySoxy)~1 -(45 pheny!piperazin-1-yl)propan-1-one.

Examples of GnRH analogues suitable for use as a TM in the present invention include those known from, for example, EP171477, WO96/033729, WO92/022322, WO92/013883, and WO91/05563, each of which is herein incorporated by reference thereto. Specific examples comprise:

(NAcDQal¹,DPtf²,DPAI³,cjsPzACAIa⁵,DPicLys⁶,DAIa¹⁰)LHRH;

(NAcDNal¹,DpCIPhe²,DPal³,cjsPzACAIa⁵,DNicLys⁶,ILys⁸,DAIa¹⁰)LHRH; (NAcDNal¹,DpCIPhe²,DPal³,Thr⁴,PicLys⁵,DPicLys⁶,ILys⁸,DAIa¹⁰)LHRH; (NAcDNal¹,DpCIPhe²,DPal³,PicLys⁵,DPicLys⁶,Thr⁷,ILys⁸,DAIa¹⁰)LHRH;

(NapDThr¹, DpCIPhe², DPal³, PicLys⁵,DPicLys⁶,ILys⁸,DAIa¹⁰)LHRH;

(NAcDNal¹ , DpCIPhe²,DPal³,NicLys⁵,DNicl_ys⁶,Thr⁷,ILys⁸,DAIa¹⁰)LHRH; (NAcDNal¹ , DpCIPhe²,DPal³,Thr⁴NicLys⁵,DNicLys⁶,Thr⁷,ILys⁸,DAIa¹⁰)LHRH; (NAcDNal¹,DpCIPhe²,DPal³,PicLys⁵,D(PicSar)Lys⁶,ILys⁸,DAIa¹⁰)LHRH’ (NAcDNal¹,DpCIPhe²,DPal³,D(PicSar)Lys⁶,ILys⁸,DAIa¹⁰)LHRH;

(NAcDNal¹,DpCIPhe²,DPal³,PicLys⁵,D(6ANic)Lys⁶,ILys⁸,DAIa¹⁰)LHRH; (NAcDNal¹,DpCIPhe²,DPal³,PicLys⁵,D(6ANic)0rn⁶,ILys⁸,DAIa¹°)LHRH; (NAcDQal¹ ,DCpa²,DPal³,cisPzACAIa⁵,DPicLys⁶,NLeu⁷,ILys⁸,DAIa¹⁰)LHRH; (NAcDNal¹,DCpa²,DPal³ DPicLys⁵,DAPhe(PicSar)^p,ILys⁸,DAIa^l0)LHRH; (NAcDQal¹ ,DCpa²,DPal³,PicLys⁵,DPal⁶,ILys⁸,DAIa¹ °)LHRH;

(NAcDNal¹ , DCpa²,DPal³,PicLys⁵,DOrn(ACyp)⁶,ILys⁸,DAIa¹⁰)LHRH; N-acetylD-beta-NaS-D-Phe-D-Phe-Ser-Tyr-D-Lys(cyclo-pentyi)-Phe-Arg-Pro-D-AiaΝΗξ; N-acetyi-D-0-Nai-D-Phe-D-Phe-Ser-Tyr-D-Lys(cyclopentyi)-PheLys(cyclopentyi)-Pro-D-Aia-NH2; N-acetyi-D-beta-Nal-D-Phe-D-Phe-Ser-TyrD-Arg-Phe-OsopropyQD-Lys-Pro-D-Ala-NH?;

N-acety1-D-beta-Nal-D-Phe-D-Phe-Ser-Tyr-D-Lys(benzyl)-Phe-Arg-Pro-DAia-NH^: N-acetyl-D-beta-Nai-D-Phe-D-Phe-Ser-Tyr-D-Lys(CI-benzyl)-PheArg-Pro-D-Ala-Nl-b; N-acety!-D-beta-Nal-D-Phe-D-Phe-Ser-Tyr-D-Lys(heptyi)Phe-Arg-Pro-D-Ala-NHs; N-acety!-D-beta-Nal~D-Phe-D-Phe-Ser-Tyr-D-Arg.Phe-Lys-(t-butylmethyi)-Pro-D-Afa-NH2; N-acetyl-D-beta-Nai-D-Phe-D-Phe-492015268754 16 Dec 2015

Ser-Tyr-D-Arg-Phe-Lys-(4-methyi-benzyl)-Pra-D-Aia-NH₂; N-acetyl-D-betaNai-D-Phe-D-Phe-Ser-Tyr-D-Arg-Phe-Lys-(benzyl)-Pro-D-Aia-NH₂; N-acetyiD-bete-Nai-D-p-CI-Phe-D-Trp-Ser-Tyr-DP-NH₂Phe~Phe-(isopropyl}Lys-ProD-Aia-NHs; N-acetyl-D-beta-Nal-D-Phe-D-Phe-Ser-Tyr-D-Lys(heptyi)-Phe5 Lys-(heptyl)-Pro-D-A!a-NH₂; N-acetyi-D-3-Na!-D-Phe-D-Phe-Ser-Tyr-D-Lys(1 butyipentyi)-Phe-Lys(1-buty!pentyi)-Arg-Pro-D~Aia-NH₂.

The polypeptides of the present invention lack a functional He domain of a clostridial neurotoxin. Accordingly, said polypeptides are not able to bind rat synaptosomal membranes (via a clostridial H_c component) in binding assays as described in Shone et al. (1985) Eur. J. Biochem. 151, 75-82. In a preferred embodiment, the polypeptides preferably lack the last 50 C-terminal amino acids of a clostridial neurotoxin holotoxin. In another embodiment, the polypeptides preferably lack the last 100, preferably the last 150, more preferably the last 200, particularly preferably the last 250, and most preferably the last 300 C-terminal amino acid residues of a clostridial neurotoxin holotoxin. Alternatively, the Hc binding activity may be negated/ reduced by mutagenesis - by way of example, referring to BoNT/ A for convenience, modification of one or two amino acid residue mutations (W1266 to L and Y1267 to F) in the ganglioside binding pocket causes the H_c region to lose its receptor binding function. Analogous mutations may be made to nonserotype A clostridial peptide components, e.g. a construct based on botulinum B with mutations (W1262 to L and Y1263 to F) or botulinum E (W1224 to L and Y1225 to F). Other mutations to the active site achieve the same ablation of H_c receptor binding activity, e.g. Y1267S in botulinum type A toxin and the corresponding highly conserved residue in the other clostridial neurotoxins. Details of this and other mutations are described in Rummel et al (2004) (Molecular Microbiol. 51:631-634), which is hereby incorporated by reference thereto.

In another embodiment, the polypeptides of the present invention lack a functional H_c domain of a clostridial neurotoxin and also lack any functionally equivalent TM. Accordingly, said polypeptides lack the natural binding function of a clostridial neurotoxin and are not able to bind rat synaptosomal

-502015268754 16 Dec 2015 membranes (via a clostridial H_c component, or via any functionally equivalent TM) in binding assays as described in Shone et al. (1985) Eur. J. Biochem. 151,75-82.

The He peptide of a native clostridial neurotoxin comprises approximately 400-440 amino acid residues, and consists of two functionally distinct domains of approximately 25kDa each, namely the N-terminal region (commonly referred to as the H_Cn peptide or domain) and the C-terminal region (commonly referred to as the H_Cc peptide or domain). This fact is confirmed by the following publications, each of which is herein incorporated in its entirety by reference thereto: Umland TC (1997) Nat. Struct. Biol. 4: 788-792; Herreros J (2000) Biochem. J. 347: 199-204; Halpern J (1993) J. Biol. Chem. 268: 15, pp. 11188-11192; Rummel A (2007) PNAS 104: 359-364; Lacey DB (1998) Nat. Struct. Biol. 5: 898-902; Knapp (1998) Am. Cryst. Assoc. Abstract

Papers 25: 90; Swaminathan and Eswaramoorthy (2000) Nat. Struct. Biol. 7: 1751-1759; and Rummel A (2004) Mol. Microbiol. 51(3), 631-643. Moreover, it has been well documented that the C-terminal region (Hcc), which constitutes the C-terminal 160-200 amino acid residues, is responsible for binding of a clostridial neurotoxin to its natural cell receptors, namely to nerve terminals at the neuromuscular junction - this fact is also confirmed by the above publications. Thus, reference throughout this specification to a clostridial heavy-chain lacking a functional heavy chain H_c peptide (or domain) such that the heavy-chain is incapable of binding to cell surface receptors to which a native clostridial neurotoxin binds means that the clostridial heavy-chain simply lacks a functional H_Cc peptide. In other words, the H_Cc peptide region is either partially or wholly deleted, or otherwise modified (e.g. through conventional chemical or proteolytic treatment) to inactivate its native binding ability for nerve terminals at the neuromuscular junction.

Thus, in one embodiment, a clostridial H_N peptide of the present invention lacks part of a C-terminal peptide portion (H_Cc) of a clostridial neurotoxin and thus lacks the He binding function of native clostridial neurotoxin. By way of example, in one embodiment, the C-terminally extended clostridial Hn peptide lacks the C-terminal 40 amino acid residues, or the C-terminal 60 amino acid

-51 2015268754 16 Dec 2015 residues, or the C-terminal 80 amino acid residues, or the C-terminai 100 amino acid residues, or the C-terminal 120 amino acid residues, or the Cterminal 140 amino acid residues, or the C-terminal 150 amino acid residues, or the C-terminal 160 amino acid residues of a clostridial neurotoxin heavy5 chain. In another embodiment, the clostridial Hn peptide of the present invention lacks the entire C-terminal peptide portion (H_Cc) of a clostridial neurotoxin and thus lacks the H_c binding function of native clostridial neurotoxin. By way of example, in one embodiment, the clostridial H_N peptide lacks the C-terminal 165 amino acid residues, or the C-terminal 170 amino acid residues, or the C-terminal 175 amino acid residues, or the C-terminal 180 amino acid residues, or the C-terminal 185 amino acid residues, or the Cterminal 190 amino acid residues, or the C-terminal 195 amino acid residues of a clostridial neurotoxin heavy-chain. By way of further example, the clostridial Hn peptide of the present invention lacks a clostridial H_Cc reference sequence selected from the group consisting of:

Botulinum type A neurotoxin - amino acid residues (Y1111 -L1296) Botulinum type B neurotoxin - amino acid residues (Y1098-E1291) Botulinum type C neurotoxin - amino acid residues (Y1112-E1291)

Botulinum type D neurotoxin - amino acid residues (Y1099-E1276)

Botulinum type E neurotoxin - amino acid residues (Y1086-K1252) Botulinum type F neurotoxin - amino acid residues (Y1106-E1274) Botulinum type G neurotoxin - amino acid residues (Y1106-E1297) Tetanus neurotoxin - amino acid residues (Y1128-D1315).

The above-identified reference sequences should be considered a guide as slight variations may occur according to sub-serotypes.

The protease of the present invention embraces all non-cytotoxic proteases that are capable of cleaving one or more proteins of the exocytic fusion apparatus in eukaryotic cells.

The protease of the present invention is preferably a bacterial protease (or fragment thereof). More preferably the bacterial protease is selected from the

-522015268754 16 Dec 2015 genera Clostridium or Neisseria/ Streptococcus (e.g. a clostridial L-chain, or a neisserial IgA protease preferably from N. gonorrhoeae orS. pneumoniae).

The present invention also embraces variant non-cytotoxic proteases (ie. 5 variants of naturally-occurring protease molecules), so long as the variant proteases still demonstrate the requisite protease activity. By way of example, a variant may have at least 70%, preferably at least 80%, more preferably at least 90%, and most preferably at least 95 or at least 98% amino acid sequence homology with a reference protease sequence. Thus, the term variant includes non-cytotic proteases having enhanced (or decreased) endopeptidase activity - particular mention here is made to the increased K_Cat/K_m of BoNT/A mutants Q161A, E54A, and K165L see Ahmed, S.A. (2008) Protein J. DOI 10.1007/s10930-007-9118-8, which is incorporated by reference thereto. The term fragment, when used in relation to a protease, typically means a peptide having at least 150, preferably at least 200, more preferably at least 250, and most preferably at least 300 amino acid residues of the reference protease. As with the TM ‘fragment’ component (discussed above), protease ‘fragments’ of the present invention embrace fragments of variant proteases based on a reference sequence.

The protease of the present invention preferably demonstrates a serine or metalloprotease activity (e.g. endopeptidase activity). The protease is preferably specific for a SNARE protein (e.g. SNAP-25, synaptobrevin/VAMP, or syntaxin).

Particular mention is made to the protease domains of neurotoxins, for example the protease domains of bacterial neurotoxins. Thus, the present invention embraces the use of neurotoxin domains, which occur in nature, as well as recombinantly prepared versions of said naturally-occurring neurotoxins.

Exemplary neurotoxins are produced by Clostridia, and the term clostridial neurotoxin embraces neurotoxins produced by C. tetani (TeNT), and by C. botulinum (BoNT) serotypes A-G, as well as the closely related BoNT-like -532015268754 16 Dec 2015 neurotoxins produced by C. baratii and C. butyricum. The above-mentioned abbreviations are used throughout the present specification. For example, the nomenclature BoNT/A denotes the source of neurotoxin as BoNT (serotype A). Corresponding nomenclature applies to other BoNT serotypes.

BoNTs are the most potent toxins known, with median lethal dose (LD50) values for mice ranging from 0.5 to 5 ng/kg depending on the serotype. BoNTs are adsorbed in the gastrointestinal tract, and, after entering the general circulation, bind to the presynaptic membrane of cholinergic nerve terminals and prevent the release of their neurotransmitter acetylcholine. BoNT/B, BoNT/D, BoNT/F and BoNT/G cleave synaptobrevin/vesicleassociated membrane protein (VAMP); BoNT/C, BoNT/A and BoNT/E cleave the synaptosomal-associated protein of 25 kDa (SNAP-25); and BoNT/C cleaves syntaxin.

BoNTs share a common structure, being di-chain proteins of -150 kDa, consisting of a heavy chain (H-chain) of -100 kDa covalently joined by a single disulphide bond to a light chain (L-chain) of -50 kDa. The H-chain consists of two domains, each of -50 kDa. The C-terminal domain (H_c) is required for the high-affinity neuronal binding, whereas the N-terminal domain (Hn) is proposed to be involved in membrane translocation. The L-chain is a zinc-dependent metalloprotease responsible for the cleavage of the substrate SNARE protein.

The term L-chain fragment means a component of the L-chain of a neurotoxin, which fragment demonstrates a metalloprotease activity and is capable of proteolytically cleaving a vesicle and/or plasma membrane associated protein involved in cellular exocytosis.

Examples of suitable protease (reference) sequences include:

Botulinum type A neurotoxin - amino acid residues (1-448)

Botulinum type B neurotoxin - amino acid residues (1-440)

Botulinum type C neurotoxin - amino acid residues (1 -441)

-542015268754 16 Dec 2015

Botulinum type D neurotoxin - amino acid residues (1-445)

Botulinum type E neurotoxin - amino acid residues (1-422)

Botulinum type F neurotoxin - amino acid residues (1-439)

Botulinum type G neurotoxin - amino acid residues (1 -441)

Tetanus neurotoxin - amino acid residues (1-457)

IgA protease - amino acid residues (1-959)* * Pohlner, J. et al. (1987). Nature 325, pp. 458-462, which is hereby incorporated by reference thereto.

The above-identified reference sequence should be considered a guide as slight variations may occur according to sub-serotypes. By way of example, US 2007/0166332 (hereby incorporated by reference thereto) cites slightly different clostridial sequences:

Botulinum type A neurotoxin - amino acid residues (M1-K448)

Botulinum type B neurotoxin - amino acid residues (M1-K441)

Botulinum type C neurotoxin - amino acid residues (M1-K449)

Botulinum type D neurotoxin - amino acid residues (M1-R445)

Botulinum type E neurotoxin - amino acid residues (M1-R422)

Botulinum type F neurotoxin - amino acid residues (M1-K439)

Botulinum type G neurotoxin - amino acid residues (M1-K446)

Tetanus neurotoxin - amino acid residues (M1-A457)

A variety of clostridial toxin fragments comprising the light chain can be useful in aspects of the present invention with the proviso that these light chain fragments can specifically target the core components of the neurotransmitter release apparatus and thus participate in executing the overall cellular mechanism whereby a clostridial toxin proteolytically cleaves a substrate. The light chains of clostridial toxins are approximately 420-460 amino acids in length and comprise an enzymatic domain. Research has shown that the entire length of a clostridial toxin light chain is not necessary for the enzymatic activity of the enzymatic domain. As a non-limiting example, the first eight amino acids of the BoNT/A light chain are not required for enzymatic activity.

-552015268754 16 Dec 2015

As another non-limiting example, the first eight amino acids of the TeNT light chain are not required for enzymatic activity. Likewise, the carboxyl-terminus of the light chain is not necessary for activity. As a non-limiting example, the last 32 amino acids of the BoNT/A light chain (residues 417-448) are not required for enzymatic activity. As another non-limiting example, the last 31 amino acids of the TeNT light chain (residues 427-457) are not required for enzymatic activity. Thus, aspects of this embodiment can include clostridial toxin light chains comprising an enzymatic domain having a length of, for example, at least 350 amino acids, at least 375 amino acids, at least 400 amino acids, at least 425 amino acids and at least 450 amino acids. Other aspects of this embodiment can include clostridial toxin light chains comprising an enzymatic domain having a length of, for example, at most 350 amino acids, at most 375 amino acids, at most 400 amino acids, at most 425 amino acids and at most 450 amino acids.

The polypeptides of the present invention, especially the protease component thereof, may be PEGylated - this may help to increase stability, for example duration of action of the protease component. PEGylation is particularly preferred when the protease comprises a BoNT/A, B or Ci protease.

PEGylation preferably includes the addition of PEG to the N-terminus of the protease component. By way of example, the N-terminus of a protease may be extended with one or more amino acid (e.g. cysteine) residues, which may be the same or different. One or more of said amino acid residues may have its own PEG molecule attached (e.g. covalently attached) thereto. An example of this technology is described in W02007/104567, which is incorporated in its entirety by reference thereto.

A Translocation Domain is a molecule that enables translocation of a protease into a target cell such that a functional expression of protease activity occurs within the cytosol of the target cell. Whether any molecule (e.g. a protein or peptide) possesses the requisite translocation function of the present invention may be confirmed by any one of a number of conventional assays.

For example, Shone C. (1987) describes an in vitro assay employing -562015268754 16 Dec 2015 liposomes, which are challenged with a test molecule. Presence of the requisite translocation function is confirmed by release from the liposomes of K⁺ and/ or labelled NAD, which may be readily monitored [see Shone C. (1987) Eur. J. Biochem; vol. 167(1): pp. 175-180].

A further example is provided by Blaustein R. (1987), which describes a simple in vitro assay employing planar phospholipid bilayer membranes. The membranes are challenged with a test molecule and the requisite translocation function is confirmed by an increase in conductance across said membranes [see Blaustein (1987) FEBS Letts; vol. 226, no. 1: pp. 115-120].

Additional methodology to enable assessment of membrane fusion and thus identification of Translocation Domains suitable for use in the present invention are provided by Methods in Enzymology Vol 220 and 221,

Membrane Fusion Techniques, Parts A and B, Academic Press 1993.

The present invention also embraces variant translocation domains, so long as the variant domains still demonstrate the requisite translocation activity. By way of example, a variant may have at least 70%, preferably at least 80%, more preferably at least 90%, and most preferably at least 95% or at least 98% amino acid sequence homology with a reference translocation domain. The term fragment, when used in relation to a translocation domain, means a peptide having at least 20, preferably at least 40, more preferably at least 80, and most preferably at least 100 amino acid residues of the reference translocation domain. In the case of a clostridial translocation domain, the fragment preferably has at least 100, preferably at least 150, more preferably at least 200, and most preferably at least 250 amino acid residues of the reference translocation domain (eg. Hn domain). As with the TM 'fragment' component (discussed above), translocation 'fragments’ of the present invention embrace fragments of variant translocation domains based on the reference sequences.

The Translocation Domain is preferably capable of formation of ion-permeable pores in lipid membranes under conditions of low pH. Preferably it has been

-572015268754 16 Dec 2015 found to use only those portions of the protein molecule capable of poreformation within the endosomal membrane.

The Translocation Domain may be obtained from a microbial protein source, 5 in particular from a bacterial or viral protein source. Hence, in one embodiment, the Translocation Domain is a translocating domain of an enzyme, such as a bacterial toxin or viral protein.

It is well documented that certain domains of bacterial toxin molecules are capable of forming such pores. It is also known that certain translocation domains of virally expressed membrane fusion proteins are capable of forming such pores. Such domains may be employed in the present invention.

The Translocation Domain may be of a clostridial origin, such as the H_N domain (or a functional component thereof). H_N means a portion or fragment of the H-chain of a clostridial neurotoxin approximately equivalent to the amino-terminal half of the H-chain, or the domain corresponding to that fragment in the intact H-chain. The H-chain lacks the natural binding function of the He component of the H-chain. In this regard, the H_c function may be removed by deletion of the He amino acid sequence (either at the DNA synthesis level, or at the post-synthesis level by nuclease or protease treatment). Alternatively, the H_c function may be inactivated by chemical or biological treatment. Thus, the H-chain is incapable of binding to the Binding

Site on a target cell to which native clostridial neurotoxin (i.e. holotoxin) binds.

Examples of suitable (reference) Translocation Domains include:

Botulinum type A neurotoxin - amino acid residues (449-871)

Botulinum type B neurotoxin - amino acid residues (441-858)

Botulinum type C neurotoxin - amino acid residues (442-866)

Botulinum type D neurotoxin - amino acid residues (446-862)

Botulinum type E neurotoxin - amino acid residues (423-845)

Botulinum type F neurotoxin - amino acid residues (440-864)

-582015268754 16 Dec 2015

Botulinum type G neurotoxin - amino acid residues (442-863)

Tetanus neurotoxin - amino acid residues (458-879)

The above-identified reference sequence should be considered a guide as 5 slight variations may occur according to sub-serotypes. By way of example,

US 2007/0166332 (hereby incorporated by reference thereto) cites slightly different clostridial sequences:

Botulinum type A neurotoxin - amino acid residues (A449-K871)

Botulinum type B neurotoxin - amino acid residues (A442-S858)

Botulinum type C neurotoxin - amino acid residues (T450-N866)

Botulinum type D neurotoxin - amino acid residues (D446-N862)

Botulinum type E neurotoxin - amino acid residues (K423-K845)

Botulinum type F neurotoxin - amino acid residues (A440-K864)

Botulinum type G neurotoxin - amino acid residues (S447-S863)

Tetanus neurotoxin - amino acid residues (S458-V879)

In the context of the present invention, a variety of Clostridial toxin H_N regions comprising a translocation domain can be useful in aspects of the present invention with the proviso that these active fragments can facilitate the release of a non-cytotoxic protease (e.g. a clostridial L-chain) from intracellular vesicles into the cytoplasm of the target cell and thus participate in executing the overall cellular mechanism whereby a clostridial toxin proteolytically cleaves a substrate. The H_N regions from the heavy chains of Clostridial toxins are approximately 410-430 amino acids in length and comprise a translocation domain. Research has shown that the entire length of a H_N region from a Clostridial toxin heavy chain is not necessary for the translocating activity of the translocation domain. Thus, aspects of this embodiment can include clostridial toxin Hn regions comprising a translocation domain having a length of, for example, at least 350 amino acids, at least 375 amino acids, at least 400 amino acids and at least 425 amino acids. Other aspects of this embodiment can include clostridial toxin H_N regions comprising translocation domain having a length of, for example, at most 350 amino acids, at most 375 amino acids, at most 400 amino acids and at most 425

-592015268754 16 Dec 2015 amino acids.

For further details on the genetic basis of toxin production in Clostridium botulinum and C. tetani, we refer to Henderson ef al (1997) in The Clostridia:

Molecular Biology and Pathogenesis, Academic press.

The term H_N embraces naturally-occurring neurotoxin Hn portions, and modified H_N portions having amino acid sequences that do not occur in nature and/ or synthetic amino acid residues, so long as the modified H_N portions still demonstrate the above-mentioned translocation function.

Alternatively, the Translocation Domain may be of a non-clostridial origin. Examples of non-clostridial (reference) Translocation Domain origins include, but not be restricted to, the translocation domain of diphtheria toxin [O=Keefe et al., Proc. Natl. Acad. Sci. USA (1992) 89, 6202-6206; Silverman ef al., J. Biol. Chem. (1993) 269, 22524-22532; and London, E. (1992) Biochem. Biophys. Acta., 1112, pp.25-51], the translocation domain of Pseudomonas exotoxin type A [Prior et al. Biochemistry (1992) 31, 3555-3559], the translocation domains of anthrax toxin [Blanke et al. Proc. Natl. Acad. Sci.

USA (1996) 93, 8437-8442], a variety of fusogenic or hydrophobic peptides of translocating function [Plank et al. J. Biol. Chem. (1994) 269, 12918-12924; and Wagner ef al (1992) PNAS, 89, pp.7934-7938], and amphiphilic peptides [Murata et al (1992) Biochem., 31, pp.1986-1992]. The Translocation Domain may mirror the Translocation Domain present in a naturally-occurring protein, or may include amino acid variations so long as the variations do not destroy the translocating ability of the Translocation Domain.

Particular examples of viral (reference) Translocation Domains suitable for use in the present invention include certain translocating domains of virally expressed membrane fusion proteins. For example, Wagner et al. (1992) and Murata ef al. (1992) describe the translocation (i.e. membrane fusion and vesiculation) function of a number of fusogenic and amphiphilic peptides derived from the N-terminal region of influenza virus haemagglutinin. Other virally expressed membrane fusion proteins known to have the desired

- 602015268754 16 Dec 2015 translocating activity are a translocating domain of a fusogenic peptide of Semliki Forest Virus (SFV), a translocating domain of vesicular stomatitis virus (VSV) glycoprotein G, a translocating domain of SER virus F protein and a translocating domain of Foamy virus envelope glycoprotein. Virally encoded

Aspike proteins have particular application in the context of the present invention, for example, the E1 protein of SFV and the G protein of the G protein of VSV.

Use of the (reference) Translocation Domains listed in Table (below) includes 10 use of sequence variants thereof. A variant may comprise one or more conservative nucleic acid substitutions and/ or nucleic acid deletions or insertions, with the proviso that the variant possesses the requisite translocating function. A variant may also comprise one or more amino acid substitutions and/ or amino acid deletions or insertions, so long as the variant possesses the requisite translocating function.

Translocation Domain source	Amino acid residues	References
Diphtheria toxin	194-380	Silverman et al., 1994, J. Biol. Chem. 269, 22524-22532 London E., 1992, Biochem. Biophys. Acta., 1113, 25-51
Domain II of pseudomonas exotoxin	405-613	Prior etal., 1992, Biochemistry 31,3555-3559 Kihara & Pastan, 1994, Bioconj Chem. 5, 532-538
Influenza virus haemagglutinin	GLFGAIAGFIENGWE GMIDGWYG, and Variants thereof	Plank etal., 1994, J. Biol. Chem. 269, 12918-12924 Wagner et al., 1992, PNAS, 89, 7934-7938 Murata etal, 1992, Biochemistry 31, 1986-1992

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Translocation Domain source	Amino acid residues	References
Semliki Forest virus fusogenic protein	Translocation domain	Kielian etal., 1996, J Cell Biol. 134(4), 863-872
Vesicular Stomatitis virus glycoprotein G	118-139	Yao etal., 2003, Virology 310(2), 319-332
SER virus F protein	Translocation domain	Seth ef a/., 2003, J Virol 77(11) 6520-6527
Foamy virus envelope glycoprotein	Translocation domain	Picard-Maureau et al., 2003, J Virol. 77(8), 4722-4730

The polypeptides of the present invention may further comprise a translocation facilitating domain. Said domain facilitates delivery of the noncytotoxic protease into the cytosol of the target cell and are described, for example, in WO 08/008803 and WO 08/008805, each of which is herein incorporated by reference thereto.

By way of example, suitable translocation facilitating domains include an enveloped virus fusogenic peptide domain, for example, suitable fusogenic peptide domains include influenzavirus fusogenic peptide domain (eg. influenza A virus fusogenic peptide domain of 23 amino acids), alphavirus fusogenic peptide domain (eg. Semliki Forest virus fusogenic peptide domain of 26 amino acids), vesiculovirus fusogenic peptide domain (eg. vesicular stomatitis virus fusogenic peptide domain of 21 amino acids), respirovirus fusogenic peptide domain (eg. Sendai virus fusogenic peptide domain of 25 amino acids), morbiliivirus fusogenic peptide domain (eg. Canine distemper virus fusogenic peptide domain of 25 amino acids), avulavirus fusogenic peptide domain (eg. Newcastle disease virus fusogenic peptide domain of 25 amino acids), henipavirus fusogenic peptide domain (eg. Hendra virus fusogenic peptide domain of 25 amino acids), metapneumovirus fusogenic peptide domain (eg. Human metapneumovirus fusogenic peptide domain of 25 amino acids) or spumavirus fusogenic peptide domain such as simian

-622015268754 16 Dec 2015 foamy virus fusogenic peptide domain; or fragments or variants thereof.

By way of further example, a translocation facilitating domain may comprise a Clostridial toxin H_Cn domain or a fragment or variant thereof. In more detail, a

Clostridial toxin H_Cn translocation facilitating domain may have a length of at least 200 amino acids, at least 225 amino acids, at least 250 amino acids, at least 275 amino acids. In this regard, a Clostridial toxin H_Cn translocation facilitating domain preferably has a length of at most 200 amino acids, at most 225 amino acids, at most 250 amino acids, or at most 275 amino acids.

Specific (reference) examples include:

Botulinum type A neurotoxin - amino acid residues (872-1110)

Botulinum type B neurotoxin - amino acid residues (859-1097)

Botulinum type C neurotoxin - amino acid residues (867-1111)

Botulinum type D neurotoxin - amino acid residues (863-1098)

Botulinum type E neurotoxin - amino acid residues (846-1085)

Botulinum type F neurotoxin - amino acid residues (865-1105)

Botulinum type G neurotoxin - amino acid residues (864-1105)

Tetanus neurotoxin - amino acid residues (880-1127)

The above sequence positions may vary a little according to serotype/ subtype, and further examples of suitable (reference) Clostridial toxin H_Cn domains include:

Botulinum type A neurotoxin - amino acid residues (874-1110)

Botulinum type B neurotoxin - amino acid residues (861-1097)

Botulinum type C neurotoxin - amino acid residues (869-1111)

Botulinum type D neurotoxin - amino acid residues (865-1098)

Botulinum type E neurotoxin - amino acid residues (848-1085)

Botulinum type F neurotoxin - amino acid residues (867-1105)

Botulinum type G neurotoxin - amino acid residues (866-1105)

Tetanus neurotoxin - amino acid residues (882-1127)

Any of the above-described facilitating domains may be combined with any of the previously described translocation domain peptides that are suitable for use in the present invention. Thus, by way of example, a non-clostridial

-632015268754 16 Dec 2015 facilitating domain may be combined with non-clostridial translocation domain peptide or with clostridial translocation domain peptide. Alternatively, a Clostridial toxin H_Cn translocation facilitating domain may be combined with a non-clostridal translocation domain peptide. Alternatively, a Clostridial toxin

Hcn facilitating domain may be combined or with a clostridial translocation domain peptide, examples of which include:

Botulinum type A neurotoxin - amino acid residues (449-1110)

Botulinum type B neurotoxin - amino acid residues (442-1097)

Botulinum type C neurotoxin - amino acid residues (450-1111)

Botulinum type D neurotoxin - amino acid residues (446-1098)

Botulinum type E neurotoxin - amino acid residues (423-1085)

Botulinum type F neurotoxin - amino acid residues (440-1105)

Botulinum type G neurotoxin - amino acid residues (447-1105)

Tetanus neurotoxin - amino acid residues (458-1127)

Sequence homology:

Any of a variety of sequence alignment methods can be used to determine percent identity, including, without limitation, global methods, local methods and hybrid methods, such as, e.g., segment approach methods. Protocols to determine percent identity are routine procedures within the scope of one skilled in the. Global methods align sequences from the beginning to the end of the molecule and determine the best alignment by adding up scores of individual residue pairs and by imposing gap penalties. Non-limiting methods include, e.g., CLUSTAL W, see, e.g., Julie D. Thompson et al., CLUSTAL W:

Improving the Sensitivity of Progressive Multiple Sequence Alignment Through Sequence Weighting, Position- Specific Gap Penalties and Weight Matrix Choice, 22(22) Nucleic Acids Research 4673-4680 (1994); and iterative refinement, see, e.g., Osamu Gotoh, Significant Improvement in Accuracy of Multiple Protein. Sequence Alignments by Iterative Refinement as Assessed by Reference to Structural Alignments, 264(4) J. Mol. Biol, 823-838 (1996), Local methods align sequences by identifying one or more conserved motifs shared by all of the input sequences. Non-limiting methods include, e.g., Match-box, see, e.g., Eric Depiereux and Ernest Feytmans, Match-Box: A Fundamentally New Algorithm for the Simultaneous Alignment of Several

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Protein Sequences, 8(5) CABiOS 501 -509 (1992); Gibbs sampling, see, e.g., C. E. Lawrence et ai., Detecting Subtle Sequence Signals: A Gibbs Sampling Strategy for Multiple Alignment, 262(5131 ) Science 208-214 (1993); Aiign-M, see, e.g., ivo Van Wall© et ak, Aiign-M - A New Algorithm for Multiple

Alignment of Highly Divergent Sequences, 20(9) Bioinformatics:1428-1435 (2004).

Thus, percent sequence identity is determined by conventional methods. See, for example, Altschul et al., Bull. Math. Bio. 48: 603-16, 1986 and

Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915-19, 1992. Briefly, two amino acid sequences are aligned to optimize the alignment scores using a gap opening penalty of 10, a gap extension penalty of 1, and the blosum 62 scoring matrix of Henikoff and Henikoff (ibid.) as shown below (amino acids are indicated by the standard one-letter codes).

Alignment scores for determining sequence identity

ARNDCQEGHILKMFPSTWYV A 4

R -1 5

N -2 0 6 D-2-2 1 6 C 0-3-3-3 9 Q-1 1 0 0-35

E -1 0 0 2 -4 2 5

G 0-2 0-1-3 -2 -2 6 H -2 0 1 -1 -3 0 0 -2 8 I -1 -3 -3 -3 -1 -3 -3 -4 -3 4 L -1 -2 -3 -4 -1 -2 -3 -4-3 2 4

K -1 2 0 -1 -3 1 1 -2 -1 -3 -2 5

M -1 -1 -2 -3 -1 0-2-3-2 1 2-1 5 F -2 -3 -3 -3 -2 -3 -3-3-1 0 0-3 0 6 P -1 -2 -2 -1 -3 -1 -1 -2 -2 -3 -3 -1 -2 -4 7 S 1 -1 1 0 -1 0 0 0 -1 -2 -2 0 -1 -2 -1 4

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T 0 -1 0-1-1 -1 -1 -2 -2 -1 -1 -1 -1 -2-115 W -3 -3 -4 -4 -2 -2 -3 -2 -2 -3 -2 -3 -1 1-4-3-211

Y -2 -2 -2 -3 -2 -1 -2 -3 2 -1 -1 -2 -1 3-3-2-2 2 7

V 0-3 -3 -3 -1 -2 -2 -3-3 3 1 -2 1 -1 -2 -2 0-3 -1 4

The percent identity is then calculated as:

Total number of identical matches _x 100 [length of the longer sequence plus the number of gaps introduced into the longer sequence in order to align the two sequences]

Substantially homologous polypeptides are characterized as having one or more amino acid substitutions, deletions or additions. These changes are preferably of a minor nature, that is conservative amino acid substitutions (see below) and other substitutions that do not significantly affect the folding or activity of the polypeptide; small deletions, typically of one to about 30 amino acids; and small amino- or carboxyl-terminal extensions, such as an amino20 terminal methionine residue, a small linker peptide of up to about 20-25 residues, or an affinity tag.

Conservative amino acid substitutions

Basic:	arginine lysine histidine
Acidic:	glutamic acid aspartic acid
Polar:	glutamine asparagine

Hydrophobic: leucine isoleucine valine

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Aromatic: phenylalanine tryptophan tyrosine

Small: glycine alanine serine threonine methionine

In addition to the 20 standard amino acids, non-standard amino acids (such as 4-hydroxyproline, 6-/V-methyl lysine, 2-aminoisobutyric acid, isovaline and a -methyl serine) may be substituted for amino acid residues of the polypeptides of the present invention. A limited number of non-conservative amino acids, amino acids that are not encoded by the genetic code, and unnatural amino acids may be substituted for clostridial polypeptide amino acid residues. The polypeptides of the present invention can also comprise non-naturally occurring amino acid residues.

Non-naturally occurring amino acids include, without limitation, trans-320 methylproline, 2,4-methano-proline, cis-4-hydroxyproline, trans-4-hydroxyproline, N-methylglycine, allo-threonine, methyl-threonine, hydroxyethylcysteine, hydroxyethylhomo-cysteine, nitro-glutamine, homoglutamine, pipecolic acid, tert-leucine, norvaline, 2-azaphenylalanine, 3-azaphenylalanine, 4-azaphenyl-alanine, and 4-fluorophenylalanine. Several methods are known in the art for incorporating non-naturally occurring amino acid residues into proteins. For example, an in vitro system can be employed wherein nonsense mutations are suppressed using chemically aminoacylated suppressor tRNAs. Methods for synthesizing amino acids and aminoacylating tRNA are known in the art. Transcription and translation of plasmids containing nonsense mutations is carried out in a cell free system comprising an E. coli S30 extract and commercially available enzymes and other reagents. Proteins are purified by chromatography. See, for example, Robertson et al., J. Am. Chem. Soc. 113:2722, 1991; Ellman et al., Methods Enzymol. 202:301, 1991; Chung et al., Science 259:806-9. 1993; and Chung

- 672015268754 16 Dec 2015 et al., Proc. Natl. Acad. Sci. USA 90:10145-9, 1993). In a second method, translation is carried out in Xenopus oocytes by microinjection of mutated mRNA and chemically aminoacylated suppressor tRNAs (Turcatti et al., 1 Biol. Chem. 271:19991-8, 1996). Within a third method, E. coli cells are cultured in the absence of a natural amino acid that is to be replaced (e.g., phenylalanine) and in the presence of the desired non-naturally occurring amino acid(s) (e.g., 2-azaphenylalanine, 3-azaphenylalanine, 4azaphenylalanine, or 4-fluorophenylalanine). The non-naturally occurring amino acid is incorporated into the polypeptide in place of its natural counterpart. See, Koide et al., Biochem. 33:7470-6, 1994. Naturally occurring amino acid residues can be converted to non-naturally occurring species by in vitro chemical modification. Chemical modification can be combined with site-directed mutagenesis to further expand the range of substitutions (Wynn and Richards, Protein Sci. 2:395-403, 1993).

A limited number of non-conservative amino acids, amino acids that are not encoded by the genetic code, non-naturally occurring amino acids, and unnatural amino acids may be substituted for amino acid residues of polypeptides ofthe present invention.

Essential amino acids in the polypeptides of the present invention can be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham and Wells, Science 244: 1081-5, 1989). Sites of biological interaction can also be determined by physical analysis of structure, as determined by such techniques as nuclear magnetic resonance, crystallography, electron diffraction or photoaffinity labeling, in conjunction with mutation of putative contact site amino acids. See, for example, de Vos et al., Science 255:30612, 1992; Smith et al.. J. Mol. Biol. 224:899-904. 1992; Wlodaver et al., FEBS

Lett. 309:59-64, 1992. The identities of essential amino acids can also be inferred from analysis of homologies with related components (e.g. the translocation or protease components) of the polypeptides of the present invention.

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Multiple amino acid substitutions can be made and tested using known methods of mutagenesis and screening, such as those disclosed by ReidhaarOlson and Sauer (Science 241:53-7. 1988) or Bowie and Sauer (Proc. Natl. Acad. Sci. USA 86:2152-6, 1989). Briefly, these authors disclose methods for simultaneously randomizing two or more positions in a polypeptide, selecting for functional polypeptide, and then sequencing the mutagenized polypeptides to determine the spectrum of allowable substitutions at each position. Other methods that can be used include phage display (e.g., Lowman et al., Biochem. 30:10832-7. 1991; Ladner et al., U.S. Patent No. 5,223,409; Huse,

WIPO Publication WO 92/06204) and region-directed mutagenesis (Derbyshire et al.. Gene 46:145. 1986; Ner et al., DNA 7:127, 1988).

Multiple amino acid substitutions can be made and tested using known methods of mutagenesis and screening, such as those disclosed by Reidhaar15 Olson and Sauer (Science 241:53-7, 1988) or Bowie and Sauer (Proc. Natl. Acad. Sci. USA 86:2152-6, 1989). Briefly, these authors disclose methods for simultaneously randomizing two or more positions in a polypeptide, selecting for functional polypeptide, and then sequencing the mutagenized polypeptides to determine the spectrum of allowable substitutions at each position. Other methods that can be used include phage display (e.g., Lowman et al., Biochem. 30:10832-7. 1991; Ladner et al., U.S. Patent No. 5,223,409; Huse, WIPO Publication WO 92/06204) and region-directed mutagenesis (Derbyshire et al., Gene 46:145, 1986; Ner et al.. DNA 7:127. 1988).

There now follows a brief description of the Figures, which illustrate aspects and/ or embodiments of the present invention.

Figure 1 - Purification of LHn/D-CT-CST28 fusion protein

Using the methodology outlined in Example 3, a LHn/D-CT-CST28 fusion protein was purified from E. coli BL21 (DE3) cells. Briefly, the soluble products obtained following cell disruption were applied to a nickel-charged affinity capture column. Bound proteins were eluted with 200 mM imidazole, treated with enterokinase to activate the fusion protein and then re-applied to a second nickel-charged affinity capture column. Samples from the

-692015268754 16 Dec 2015 purification procedure were assessed by SDS-PAGE. Lane 1'. First nickel chelating Sepharose column eluant, Lane 2: Second nickel chelating Sepharose column eluant under non-reducing conditions, Lane 3: Second nickel chelating Sepharose column eluant under reducing conditions, lane 4:

Molecular mass markers (kDa).

Figure 2 - Purification of LHn/A-CT-SST14 fusion protein

Using the methodology outlined in Example 3, an LHn/A-CT-SST14 fusion protein was purified from E. coli BL21 (DE3) cells. Briefly, the soluble products obtained following cell disruption were applied to a nickel-charged affinity capture column. Bound proteins were eluted with 200 mM imidazole, treated with Factor Xa to activate the fusion protein and then re-applied to a second nickel-charged affinity capture column. Samples from the purification procedure were assessed by SDS-PAGE. Lane 1: First nickel chelating

Sepharose column eluant, Lane 2: Molecular mass markers (kDa), Lanes 3-4: Second nickel chelating Sepharose column eluant under non-reducing conditions, Lanes 5-6: Second nickel chelating Sepharose column eluant under reducing conditions.

Figures 3-47 - Box Plots showing up-regulation of SNARE protein mRNA in cancer cells

Statistical analysis of the differences in SNARE expression between normal and primary cancer tissues was completed through use of Oncomine algorithms (Compendia Bioscience, Ann Arbor, Ml, USA) and gene microarray analysis tool. SNARE mRNA expression was compared to the median expression of all other genes in the respective study, for which a Normalised expression value was generated. Only studies with analysis results with P<0.05 are illustrated. In more detail, Figures 3-10 illustrate SNAP-25 expression profiles in different cancer versus non-cancer cells. Figures 11-16 illustrate syntaxin-1 expression profiles in different cancer versus non-cancer cells. Figures 17-20 illustrate syntaxin-2 expression profiles in different cancer versus non-cancer ceils. Figures 21-24 illustrate syntaxin-3 expression profiles in different cancer versus non-cancer cells. Figures 25-32 illustrate VAMP-1 expression profiles in different cancer versus non-cancer

- 702015268754 16 Dec 2015 cells. Figures 33-38 illustrate VAMP-2 expression profiles in different cancer versus non-cancer cells. Figures 39-45 illustrate VAMP-3 expression profiles in different cancer versus non-cancer cells. In all cases, the data illustrate upregulation of SNARE proteins in cancer cells. Figure 46 illustrates a statistically significant difference in syntaxin-1 mRNA expression level between patients with recurrent invasive ductal breast cancer at 5 years post diagnosis and those patients without recurrence. Figure 47 illustrates a statistically significant difference in syntaxin-1 mRNA expression level in breast cancer patients in whom a metastatic event has occurred at 5 years post diagnosis versus those without metastases. Data is presented in box-plot form (see Figure 3 for explanation) and/or as a histogram, where each bar represents the normalised gene expression level in each patient tissue sample.

Figure 48 - Profiling cell surface receptor and SNARE protein expression in Renal Cell carcinoma lines by Western blot analysis.

Protein extracts from eight human renal cell carcinoma (RCC) lines grown in vitro were prepared in standard Laemmli sample buffer (lanes 3-10). Protein extracts prepared from primary cultures of rat spinal cord neurons and dorsal root ganglia were included as controls. Proteins were separated on 10% SDS20 polyacrylamide gels and electrophoretically transferred to a nitrocellulose membrane. After blocking of the membrane, primary antisera were used to probe for each specific protein and detection was enabled by a peroxidiseconjugated anti-species IgG. The receptor proteins detected were ErbB receptor (epidermal growth factor receptor, EGF) and growth hormone releasing hormone receptor (GHRHR). The SNARE proteins tested were SNAP-25, syntaxin-2 and syntaxin-3. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a protein loading control.

Figure 49 - Detection of SNAP-25 protein cleavage in a Renal Cell carcinoma line by Western blot analysis after 24hr treatment with an EGF-LHA fusion.

Figure 49 shows the effect of 24 hour treatment of 786-0 human RCC cells with a number of EGF-liganded fusion protein molecules. In more detail, EGFLHA, at doses between 1 and 100nM, generated cleaved SNAP-25 species

- 71 2015268754 16 Dec 2015 whereas control molecules, specifically EGF-LHB (non- SNAP-25 targeting), a catalytically inactive form of EGF-LHA, EGF-LH-DE-A (referred to as ‘EGF-0’ from hereon in) and the ‘free’ EGF ligand, did not. Indicated concentrations are in nM.

Figure 50 - Inhibition of in vitro proliferation of a Renal Cell carcinoma line by an EGF-LHA fusion.

786-0 cells seeded into a cell culture vessel were counted in a pre-define region at 24 and 48 hours after treatment with 25nM of EGF-iiganded fusion proteins.

Figure 51 - inhibition of FGF-2 secretion from a Renal Cell carcinoma line by an EGF-LHA fusion.

Culture media from 786-0 cells treated for 24 hours with EGF-liganded fusion 15 proteins were analysed for Fibroblast Growth Factor-2 by standard methods.

In more detail, EGF-LHA, at doses between 1 and 50nM, demonstrated a dose-dependent decrease in FGF-2 levels present in the culture medium whereas control molecules, specifically the catalytically inactive form of EGFLHA, EGF-0, and ‘free’ EGF ligand, did not. Indicated concentrations are in nM.

Figure 52 - Inhibition of in vitro proliferation of the A498 Renal Cell carcinoma line by an EGF-LHA fusion.

Figure 52 shows the effect of an EGF-liganded fusion protein (EGFv3-LHA, 25 300nM) on cellular proliferation of A498 cell line after 48 hours. Cells were stained using the tetrazolium salt WST as per standard protocols. The unliganded molecule LHA was included as a control.

Figure 53 - Inhibition of in vitro proliferation of the ACHN Renal Cell 30 carcinoma line by an EGF-LHA fusion.

Figure 53 shows the effect of an EGF-liganded fusion protein (EGFv3-LHA, at 300nM) on cellular proliferation of ACHN cell line after 48 hours. Cells were stained using the tetrazolium salt WST as per standard protocols. The unliganded molecule LHA was included as a control.

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Figure 54 - Inhibition of in vitro secretion of gastrin releasing peptide (GRP) from the small cell lung cancer cel, line DMS-53 by an EGF-LHA fusion.

Figure 54 shows the effect of an EGF-liganded fusion protein (EGFv3-LHA, at 150nM) on GRP secretion from the DMS-53 cell line after treatment for 24hr. Medium removed from the cells were analysed 48 hours after removal of fusion proteins. The catalytically inactive form of EGF-LHA, EGF-0, was included as a control, as was cycloheximide, a general protein synthesis inhibitor.

Nomenclature
SST	somatostatin
TGF(A)	transforming growth factor (alpha)
GHRL	ghrelin
LEP	leptin
ET(A)	endothelin-1
FLT	vascular endothelial growth factor receptor
CHRN(D)	acetylcholine receptor (subunit delta)
EPHA	ephrin type-A receptor
EFNA	ephrin-A
DLK1	delta-like protein 1
JAG	jagged protein
NRG	neuregulin
G-CSF	granulocyte colony-stimulating factor
AMF	autocrine motility factor
NMB	neuromedin-B
CCK	cholecystokinin
PDGF	platelet-derived growth factor
ADM	adrenomedullin
GDNF	glial cell line-derived neurotrophic factor
TrkA	high affinity nerve growth factor
FSH	follicle-stimulating hormone
CXCR	C-X-C chemokine receptor

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CRLR calcitonin-receptor-like receptor

PDF prostate differentiation factor

MCP monocyte chemotactic protein

KGF keratinocyte growth factor

FLK1 vascular endothelial growth factor receptor 2

PDGFR platelet-derived growth factor receptor

NOTCH neurogenic locus notch homolog protein

DLL delta-like protein

GHS growth hormone secretagogue c-MET hepatocyte growth factor c-kit mast/stem cell growth factor

MGSA/GRO melanoma growth stimulatory activity/growth related gene

BCGF B-cell growth factor

GnRH gonadotropin-releasing hormone receptor

Ang-2 angiopoietin-2

FGF fibroblast growth factor

ErbB epidermal growth factor receptor family member

VIPR vasoactive intestinal polypeptide receptor

BRS bombesin receptor subtype

GRP gastrin releasing peptide

LIF leukaemia inhibitory factor

GHRH growth hormone-releasing hormone

IGF insulin-like growth factor

CRHR-2 corticotropin releasing factor receptor-2

BB bombesin

GH growth hormone

IL interleukin

VEGF vascular endothelial growth factor

ACH acetylcholine

CST cortistatin

VPAC vasoactive intestinal peptide receptor

GRPR gastrin releasing peptide receptor

CTR calcitonin binding receptor

EPO erythropoietin

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ΗΒ-EGF heparin-binding EGF-like growth factor

HGF/SF hepatocyte growth factor/scatter factor

SDF-1 stromal cell-derived factor 1

CXCL12 chemokine (C-X-C motif) ligand 12 (SDF-1)

TNF tumour necrosis factor

PGF placental growth factor

Gran4 granulin-4

TIE2 angiopoietin receptor-2

LH luteinising hormone

CCL CC chemokine ligand

NT neurotrophin

NTAK neuregulin-2

BAFF B-cell activating factor

GM-CSF granulocyte-macrophage colony stimulating factor

NGF nerve growth factor

PACAP pituitary adenylate cyclase-activating peptide

OB leptin

NRP neuropilin receptor

Summary of Examples

Example 1 Preparation of a LHA backbone construct Example 2 Construction of LHD-CT-CST28

Example 3 Expression and purification of a LHD-CT-CST28 fusion protein Example 4 Construction of LHA-CP-EGF

Example 5 Expression and purification of a LHA-CP-EGF fusion protein Example 6 Chemical conjugation of LHn/A to GnRH TM Example 7 Method for treating colorectal cancer Example 8 Method for treating breast cancer Example 9 Method for treating prostate cancer

Example 10 Method for treating lung carcinoid tumours Example 11 Method for treating bladder cancer Example 12 Method for treating small cell lung cancer Example 13 Method for treating prostate cancer Example 14 Method for treating cervical cancer

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Example 15 Method for treating leukaemia Example 16 Method for treating small cell lung cancer Example 17 Method for treating pancreatic cancer Example 18 Method for treating metastatic bone cancer

Example 19 Method for treating metastatic small cell lung cancer in the brain Example 20 Method for treating bowel cancer Example 21 Method for treating chronic lymphocytic leukaemia Example 22 Method for treating liver cancer Example 23 Method for treating Hodgkin’s lymphoma

Example 24 Method for treating renal cancer Example 25 Method for treating skin cancer Example 26 Method for treating oropharyngeal cancer Example 27 Method for treating myeloma cancer Example 28 Method for treating soft tissue sarcoma cancer

Example 29 Method for treating gastric cancer Example 30 Method for treating testicular cancer Example 31 Method for treating uterine cancer Example 32 Method for treating Karposi sarcoma Example 33 Method for treating primary brain cancer

Example 34 Method for treating rectal cancer

Example 35 Assessment of proliferation changes, inhibition of cellular secretion and concomitant SNARE cleavage after treatment of in vitro cultured renal cancer cell lines with a polypeptide of the present invention.

Summary of SEQ ID NOs

Where an initial Met amino acid residue or a corresponding initial codon is indicated in any of the following SEQ ID NOs, said residue/ codon is optional.

1. DNA sequence of LH_N/A 30 2. DNA sequence of LH_N/B

3. DNA sequence of LHn/C

4. DNA sequence of LH_N/D

5. DNA sequence of the CT-CST28 linker

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6. DNA sequence of the LHD-CT-CST28 fusion

7. Protein sequence of the LHD-CT-CST28 fusion

8. DNA sequence of the CP-EGF linker

9. DNA sequence of the LHA-CP-EGF fusion

10. Protein sequence of the LHA-CP-EGF fusion

11. Protein sequence of LH_N/A

12. Protein sequence of LHn/B

13. Protein sequence of LH_N/C

14. Protein sequence of LH_N/D

15. Synthesised GnRH peptide

16. Protein sequence ofthe LHB-CT-SST28 fusion

17. Protein sequence ofthe LHA-CP-SST28 fusion

18. Protein sequence ofthe LHD-CT-EGF fusion

19. Protein sequence of the LHD-CT-VIP fusion

20. Protein sequence of the LHC-CT-IGF1 fusion

21. Protein sequence ofthe LHD-CT-IGF1 fusion

22. Protein sequence ofthe LHC-CT-VIP fusion

23. Protein sequence ofthe LHC-CT-GnRH fusion

24. Protein sequence ofthe LHD-CT-GnRH fusion

25. Protein sequence of the LHD-CT-GRP fusion

26. Protein sequence ofthe LHB-CT-GRP fusion

27. Protein sequence ofthe LHC-CT-LIF fusion

28. Protein sequence ofthe LHB-CP-LIF fusion

29. Protein sequence ofthe LHC-CT-FGF1 fusion

30. Protein sequence of the LHA-CP-FGF1 fusion .Protein sequence ofthe LHA-CT-FGF9 fusion

32. Protein sequence ofthe LHC-CP-FGF9 fusion

33. Protein sequence ofthe lgA-HNtet-CT-SST14 fusion

34. Protein sequence ofthe lgA-HNtet-CP-SST14 fusion

35.Protein sequence ofthe LHA-CT-SST14 fusion

36. Protein sequence ofthe LHA-CT-EGFv3 fusion

37. Protein sequence ofthe LHE-CT-IL6 fusion

38. Protein sequence ofthe LHB-CT-IL8 fusion

39. Protein sequence ofthe LHF-CP-GRAN4 fusion

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40. Protein sequence of the LHD-CP-TGFa fusion 41 .Protein sequence of the LHD-CP-TGFb fusion

42. Protein sequence of the LHB-CT-TNFa fusion

43. Protein sequence of the LHD-CT-SDF1 fusion

44. Protein sequence of the LHC-CT-VEGF fusion

Examples

Example 1 Preparation of a LH_N/A backbone construct

The following procedure creates a clone for use as an expression backbone for multidomain protein expression. This example is based on preparation of a serotype A based clone (SEQ ID1), though the procedures and methods are equally applicable to all LH_N serotypes such as serotype B (SEQ ID2), serotype C (SEQ ID3) and serotype D (SEQ ID4) and other protease or translocation domains by using the appropriate published sequence for synthesis.

Preparation of cloning and expression vectors pCR 4 (Invitrogen) is the chosen standard cloning vector chosen due to the lack of restriction sequences within the vector and adjacent sequencing primer sites for easy construct confirmation. The expression vector is based on the pET (Novagen) expression vector which has been modified to contain the multiple cloning site Ndel-BamHI-Sall-Pstl-Xbal-Hindlll for construct insertion, a fragment of the expression vector has been removed to create a non25 mobilisable plasmid, a variety of different fusion tags have been inserted to increase purification options and an existing Xbal site in the vector backbone has been removed to simplify sub-cloning.

Preparation of LC/A

The DNA sequence is designed by back translation of the LC/A amino acid sequence (obtained from freely available database sources such as GenBank (accession number P10845) using one of a variety of reverse translation software tools (for example Backtranslation tool v2.0 (Entelechon)). BamHI/Sall recognition sequences are incorporated at the 5’ and 3’ ends

-782015268754 16 Dec 2015 respectively of the sequence maintaining the correct reading frame. The DNA sequence is screened (using software such as SeqBuilder, DNASTAR Inc.) for restriction enzyme cleavage sequences incorporated during the back translation. Any cleavage sequences that are found to be common to those required by the cloning system are removed by the Backtranslation tool from the proposed coding sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the %GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, September 13 2004). This optimised DNA sequence containing the LC/A open reading frame (ORF) is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Preparation of Hn/A insert

The DNA sequence is designed by back translation of the H_N/A amino acid sequence (obtained from freely available database sources such as GenBank (accession number P10845) using one of a variety of reverse translation software tools (for example Back translation tool v2.0 (Entelechon)). A Pstl restriction sequence added to the N-terminus and Xbal-stop codon-Hindlll to the C-terminus ensuring the correct reading frame in maintained. The DNA sequence is screened (using software such as SeqBuilder, DNASTAR Inc.) for restriction enzyme cleavage sequences incorporated during the back translation. Any sequences that are found to be common to those required by the cloning system are removed by the Backtranslation tool from the proposed coding sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the %GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, September 13 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Preparation of the interdomain (LC-H_N linker)

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The LC-Hn linker can be designed from first principle, using the existing sequence information for the linker as the template. For example, the serotype A linker (in this case defined as the inter-domain polypeptide region that exists between the cysteines of the disulphide bridge between LC and

Hn) has the sequence VRGIIPFKTKSLDEGYNKALNDL. This sequence information is freely available from available database sources such as GenBank (accession number P10845). For generation of a specific protease cleavage site, the native recognition sequence for Factor Xa can be used in the modified sequence VDGIITSKTKSLIEGRNKALNLQ or an enterokinase recognition sequence is inserted into the activation loop to generate the sequence VDGIITSKTKSDDDDKNKALNLQ. Using one of a variety of reverse translation software tools (for example Backtranslation tool v2.0 (Entelechon), the DNA sequence encoding the linker region is determined. BamHI/Sall and Pstl/Xbal/stop codon/HindlII restriction enzyme sequences are incorporated at either end, in the correct reading frames. The DNA sequence is screened (using software such as Seqbuilder, DNASTAR Inc.) for restriction enzyme cleavage sequences incorporated during the back translation. Any sequences that are found to be common to those required by the cloning system are removed by the Backtranslation tool from the proposed coding sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the %GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, September 13 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or SigmaGenosys) and is provided in the pCR 4 vector.

Assembly and confirmation of the backbone clone

Due to the small size, the activation linker must be transferred using a two step process. The pCR-4 linker vector is cleaved with BamHI + Sail combination restriction enzymes and the cleaved linker vector then serves as the recipient for BamHI + Sail restriction enzyme cleaved LC DNA. Once the LC encoding DNA is inserted upstream of the linker DNA, the entire LC-linker DNA fragment can then be isolated and transferred to the pET expression

- 802015268754 16 Dec 2015 vector MCS. The LC-linker is cut out from the pCR 4 cloning vector using BamHI/Pstl restriction enzymes digests. The pET expression vector is digested with the same enzymes but is also treated with antarctic phosphatase as an extra precaution to prevent re-circularisation. The LC5 linker and the pET vector backbone are gel purified and the purified insert and vector backbone are ligated together using T4 DNA ligase. The product is transformed with TOP10 cells which are then screened for LC-linker using BamHI/Pstl restriction digestion. The process is then repeated for the Hn insertion into the Pstl/HindIII restriction sites of the pET-LC-linker construct.

Screening with restriction enzymes is sufficient to ensure the final backbone is correct as all components are already sequenced confirmed during synthesis. However, during the sub-cloning of some components into the backbone, where similar size fragments are being removed and inserted, sequencing of a small region to confirm correct insertion is required.

Example 2 Construction of LH_N/D-CT-CST28

The following procedure creates a clone for use as an expression construct for multidomain fusion expression where the targeting moiety (TM) is presented C-terminally to the translocation domain. This example is based on preparation of the LHn/D-CT-CST28 fusion (SEQ ID6), though the procedures and methods are equally applicable to create other protease, translocation and TM fusions, where the TM of C-terminal to the translocation domain. In this example, a flanking 15 amino acid glycine-serine spacer is engineered into the interdomain sequence to ensure accessibility of the ligand to its receptor, but other spacers are applicable.

Preparation of spacer-CST28 insert

For presentation of a CST28 sequence at the C-terminus of the Hn domain, a

DNA sequence is designed to flank the spacer and targeting moiety (TM) regions allowing incorporation into the backbone clone (SEQ ID4). The DNA sequence can be arranged as SamHI-Sa/l-Psfl-Xbal-spacer-CST28-stop codon-/-//ridIiI (SEQ ID5). The DNA sequence can be designed using one of a variety of reverse translation software tools (for example EditSeq best E. coli

- 81 2015268754 16 Dec 2015 reverse translation (DNASTAR Inc.), or Backtranslation tool v2.0 (Entelechon)). Once the TM DNA is designed, the additional DNA required to encode the preferred spacer is created in silico. It is preferred to ensure the correct reading frame is maintained for the spacer, TM and restriction sequences and that the Xba\ sequence is not preceded by the bases TC, which would result in DAM methylation. The DNA sequence is screened for restriction sequences incorporated and any additional sequences are removed manually from the remaining sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the %GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, September 13 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Assembly and confirmation of the backbone clone

In order to create a LHn/D-CT-CST28 construct (SEQ ID6) using the backbone construct (SEQ ID4) and the newly synthesised pCR 4-spacer-TM vector encoding the CST28 TM (SEQ ID5), a one or two step method can be used; typically the two step method is used when the TM DNA is less than 100 base pairs. Using the one step method the TM can be inserted directly into the backbone construct buy cutting the pCR 4-spacer-TM vector with Xba\ and Hind\\\ restriction enzymes and inserting the TM encoding DNA fragment into a similarly cut pET backbone construct. Using the two-step method the LH_n domain is excised from the backbone clone using restriction enzymes SamHI and Xba\ and ligated into similarly digested pCR 4-spacer-TM vector. This creates an LH_N-spacer-TM ORF in pCR 4 that can be excised from the vector using restriction enzymes Bam HI and HindlW for subsequent ligation into the similarly cleaved pET expression construct. The final construct contains the LC-linker-HN-spacer-CST28 DNA (SEQ ID6) which will result in a fusion protein containing the sequence illustrated in SEQ ID7.

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Example 3 Expression and purification of a LHn/D-CT-CST28 fusion protein

This example is based on preparation of an LH_N/D protein that incorporates a CST28 TM polypeptide at the carboxyl terminus of the H_N domain (SEQ ID7), where the pET expression vector ORF also encodes a histidine purification tag. These procedures and methods are equally applicable to fusion protein sequences shown in SEQ ID 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29, 31, 33, 35, 36, 37, 38, 42, 43 or 44. Where appropriate, the activation enzyme should be selected to be compatible with the protease activation site within each sequence.

Expression of LHn/D-CT-CST28

Expression of the LH_N/D-CT-CST28 protein is achieved using the following protocol. Inoculate 100 ml of modified TB containing 0.2 % glucosamine and

30 pg/ml kanamycin in a 250 ml flask with a single colony from the LH_N/D-CTCST28 expression strain. Grow the culture at 37°C, 225 rpm for 16 hours. Inoculate 1 L of modified TB containing 0.2 % glucosamine and 30 pg/ml kanamycin in a 2 L flask with 10 ml of overnight culture. Grow cultures at 37°C until an approximate Οϋβοο nm of 0.5 is reached at which point reduce the temperature to 16°C. After 1 hour induce the cultures with 1 mM IPTG and grow at 16°C for a further 16 hours.

Purification of LHn/D-CT-CST28 protein

Defrost falcon tube containing 35 ml 50 mM HEPES pH 7.2 200 mM NaCI and approximately 10 g of E. coli BL21 (DE3) cell paste. Homogenise the cell paste (20 psi) ensuring the sample remains cool. Spin the lysed cells at 18 000 rpm, 4°C for 30 minutes. Load the supernatant onto a 0.1 M N1SO4 charged Chelating column (20-30 ml column is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCI. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. The eluted fusion protein is dialysed against 5 L of 50 mM HEPES pH 7.2 200 mM NaCI at 4°C overnight and the OD₂8o nm measured to establish the protein concentration. Add 3.2 pi enterokinase (New England Biolabs) per mg fusion protein and incubate static overnight at

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25°G. Load onto a 0.1 M N1SO4 charged Chelating column (20-30 ml column is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCI. Wash column to baseline with 50 mM HEPES pH 7.2 200 mM NaCI. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. Dialyse the eluted fusion protein against 5 L of 50 mM HEPES pH 7.2 150 mM NaCI at 4°C overnight and concentrate the fusion to about 2 mg/ml, aliquot sample and freeze at -20°C. Test purified protein using OD280, BCA and purity analysis. Figures 1 and 2 demonstrate purification of fusion proteins following this method as analysed by SDS-PAGE.

Example 4 Construction of LHn/A-CP-EGF

The following procedure creates a clone for use as an expression construct for multidomain fusion expression where the targeting moiety (TM) is presented centrally between the protease and translocation domain. This example is based on preparation of the LHn/A-CP-EGF fusion (SEQ ID9), though the procedures and methods are equally applicable to create other protease, translocation and TM fusions, where the TM is N-terminal to the translocation domain. In this example, a flanking helical spacer is engineered into the interdomain sequence to ensure accessibility of the ligand to its receptor, but other spacers are applicable.

Preparation of spacer-human EGF insert

The LC-H_n inter-domain polypeptide linker region exists between the cysteines of the disulphide bridge between LC and Hn- For insertion of a protease cleavage site, spacer and a targeting moiety (TM) region into the activation loop, one of a variety of reverse translation software tools (for example Backtranslation tool v2.0 (Entelechon) are used to determine the DNA sequence encoding the linker region. For central presentation of a TM sequence at the N-terminus of the Hn domain, a DNA sequence is designed for the spacer and targeting moiety (TM) regions allowing incorporation into the backbone clone (SEQ ID1). The DNA sequence can be arranged as BamHI-Sa/l-spacer-protease activation site-EGF-spacer-Psfl-Xdal-stop codon-/7/ndlll (SEQ ID8). Once the TM DNA is designed, the additional DNA

- 84 2015268754 16 Dec 2015 required to encode the preferred spacer is created in silico. It is preferred to ensure the correct reading frame is maintained for the spacer, TM and restriction sequences and that the Xba\ sequence is not preceded by the bases TC, which would result in DAM methylation. The DNA sequence is screened for restriction sequence incorporated and any additional sites are removed manually from the remaining sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the %GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, September 13 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Assembly and confirmation of the backbone clone

In order to create a LC-spacer-activation site-EGF-spacer-H_N construct (SEQ ID9) using the backbone construct (SEQ ID1) and the newly synthesised pCR 4-spacer-activation site-TM-spacer vector encoding the EGF TM (SEQ ID8), a one or two step method can be used; typically the two step method is used when the TM DNA is less than 100 base pairs. Using the one step method the TM linker region can be inserted directly into the backbone construct buy cutting the pCR 4- spacer-activation site-TM-spacer vector with Sa/I and Pst\ restriction enzymes and inserting the TM encoding DNA fragment into a similarly cut pET backbone construct. Using the two-step method the LC domain is excised from the backbone clone using restriction enzymes BamHI and Sa/I and ligated into similarly digested pCR 4- spacer-activation site-TMspacer vector. This creates a LC-spacer-activation site-TM-spacer ORF in pCR 4 that can be excised from the vector using restriction enzymes BamHI and Pstl for subsequent ligation into similarly pET expression construct. The final construct contains the LC-spacer-activation site-EGF-spacer-HN DNA (SEQ ID9) which will result in a fusion protein containing the sequence illustrated in SEQ ID10.

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Example 5 Expression and purification of a LH_N/A-CP-EGF fusion protein

This example is based on preparation of an LH_N/A protein that incorporates a EGF TM polypeptide into the interdomain linker region (SEQ ID10), where the pET expression vector ORF also encodes a histidine purification tag. These procedures and methods are equally applicable to the other fusion protein shown in SEQ ID 17, 28, 30, 32, 34, 39, 40 or 41. Where appropriate, the activation enzyme should be selected to be compatible with the protease activation site within each sequence.

Expression of LHn/A-CP-EGF

Expression of the LHn/A-CP-EGF protein is achieved using the following protocol. Inoculate 100 ml of modified TB containing 0.2 % glucosamine and 30 pg/ml kanamycin in a 250 ml flask with a single colony from the LHA-CP15 EGF expression strain. Grow the culture at 37°C, 225 rpm for 16 hours. Inoculate 1 L of modified TB containing 0.2 % glucosamine and 30 pg/ml kanamycin in a 2 L flask with 10 ml of overnight culture. Grow cultures at 37°C until an approximate Οϋβοο nm of 0.5 is reached at which point reduce the temperature to 16°C. After 1 hour induce the cultures with 1 mM IPTG and grow at 16°C for a further 16 hours.

Purification of LHn/A-CP-EGF protein

Defrost falcon tube containing 35 ml 50 mM HEPES pH 7.2 200 mM NaCI and approximately 10 g of E. coli BL21 (DE3) cell paste. Homogenise the cell paste (20 psi) ensuring the sample remains cool. Spin the lysed cells at 18 000 rpm, 4°C for 30 minutes. Load the supernatant onto a 0.1 M NiSO4 charged Chelating column (20-30 ml column is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCI. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. The eluted fusion protein is dialysed against 5 L of 50 mM HEPES pH 7.2 200 mM NaCI at 4°C overnight and the OD₂8o nm measured to establish the protein concentration. Add 3.2 pi enterokinase (New England Biolabs) per mg fusion protein and incubate static overnight at 25°C. Load onto a 0.1 M NiSO4 charged Chelating column (20-30 ml column

-862015268754 16 Dec 2015 is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCl. Wash column to baseline with 50 mM HEPES pH 7.2 200 mM NaCl. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. Dialyse the eluted fusion protein against 5L of 50 mM HEPES pH 7.2 150 mM NaCl at 4°C overnight and concentrate the fusion to about 2 mg/ml, aliquot sample and freeze at -20°C. Test purified protein using OD₂8o, BCA and purity analysis.

Example 6 Chemical conjugation of LHn/A to GnRH TM

The following procedure creates a chemically conjugated molecule containing the LHn/A amino acid sequence (SEQ ID11), prepared from SEQ ID1 using the production method outlined in example 3, and a GnRH peptide which has been chemically synthesised (SEQ ID15). However, the procedures and methods are equally applicable for the conjugation of other peptides to other protease/translocation domain proteins such as those containing the amino acid sequences SEQ ID12, 13 and 14.

The LHn/A protein was buffer exchanged from 50 mM Hepes 150 mM salt into PBSE (100mM 14.2g NA2HPO4, 100mM 5.85g NaCl, 1mM EDTANa₂ pH 7.5 with 1M HCI) using the Bio-rad PD10 column. This was done by washing one column volume of PBSE through the PD10 column, the protein was then added to the column until no more drops exit the end of the PD10 column. 8 mis of PBSE was then added and 0.5ml fractions are collected. The collected fractions are the measured using the A₂₈o reading and fractions containing protein are pooled. A concentration of 1.55 mg/ml of LH_N/A was obtained from the buffer exchange step and this was used to set up the following reactions:

LHn/A 1.55 mg/ml	20 mM SPDP or Sulfo-LC-SPDP
A 200 pi	0
B 200 pi	4 fold increase 0.62 pi
C 200 pi	8 fold increase 1.24 pi

- 872015268754 16 Dec 2015

Sample were left to tumble at RT for 3 hours before being passed down another PD10 column to buffer exchange into PBSE and the protein containing fractions pooled. A final concentration of 25 mM DTT was then added to derivatised protein and then the samples left at room temperature for

10 minutes. A₂₈o and A343 readings were then taken to work out the ratio of

SPDP:LH_n/A interaction and the reaction which resulted in a derivatisation ration of between 1 and 3 was used for the peptide conjugation. The SPDP reagent binds to the primary amines of the LHn/A via an Nhydroxysuccinimide (NHS) ester, leaving the sulphydryl-reactive portion to form a disulphide bond to the free SH group on the free cysteine on the synthesised peptide. In this case the peptide sequence is GnRH which has been synthesised with a cysteine contained within the peptide to allow conjugation whilst leaving the N-terminus and C-terminus of the GnRH to interact with its receptor (SEQ ID15). The SPDP-derivatised LHn/A was mixed with a 4-fold excess of the GnRH ligand and the reaction was then left at RT for 90 minutes whilst tumbling. The excess GnRH was then removed using either a PD10 column leaving LHN/A-GnRH conjugated molecule.

Example 7 - Method for treating colorectal cancer

A 62 year old man presents with a stage II colorectal cancer. To reduce and/ or to prevent metastasis he receives a direct injection of a polypeptide of the present invention (eg. botulinum type A neurotoxin protease and translocation domain and a VIP peptide). Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is optionally performed in combination with chemotherapy, and is repeated 2 months later and 4 weeks later no tumour is observable anymore with the usual detection tools (colonoscopy, CT scan, PET scan, etc.) and the level of carcinoembryonic antigen (CEA) returned to the normal.

Example 8 - Method for treating breast cancer

A 61 year old woman presents with a stage II breast cancer. To treat and/ or prevent metastasis she receives an IV injection of a polypeptide of the present invention (eg. a botulinum type C neurotoxin protease, a botulinum type C neurotoxin translocation domain and a GnRH peptide), optionally in

- 882015268754 16 Dec 2015 combination with chemotherapy. Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 6 weeks later no tumour is observable anymore with the usual detection tools (MRI, ultrasound, breast-specific positron emission tomography, mammography, Scintigraphy, etc).

Example 9 - Method for treating prostate cancer

A 77 year old man presents with a stage II prostate cancer. To treatment and/ or to prevent metastasis he receives a intravenous injection of a polypeptide of the present invention (eg. a botulinum type C neurotoxin protease, a botulinum type C neurotoxin translocation domain and an IGF-1 peptide), optionally in combination with hormone therapy. Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 8 weeks later no tumour is observable anymore with the usual detection tools (X-ray, ProstaScint scan, MRI, transrectal ultrasonography, CT scan, etc.) and the levels of PSA came back to normal.

Example 10 - Method for treating lung carcinoid tumours

A 66 year old woman presents with lung carcinoid tumours. To treat and/ or to prevent metastasis she receives an IV injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain and a bFGF-1 peptide), optionally in combination with chemotherapy. Within 4 weeks a significant decrease in the size of the tumour is observed without appearance of metastasis elsewhere.. The treatment is repeated 1 month later and 4 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan, bronchoscopy, etc.) or using the usual blood tests recommended for this cancer.

Example 11 - Method for treating bladder cancer

A 56 year old man presents with a stage II bladder cancer. To treat and/ or to prevent metastasis he receives a direct injection of a polypeptide of the present invention (eg. an IgA protease, a tetanus neurotoxin translocation domain and an IGF-1 peptide), optionally in combination with chemotherapy. Within 2

- 892015268754 16 Dec 2015 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 4 weeks later no tumour is observable anymore with the usual detection tools (colonoscopy, CT scan, PET scan, etc.).

Example 12 - Method for treating small cell lung cancer

A 79 year old man is diagnosed with a stage I small cell lung cancer. To treat and/ or to prevent metastasis he receives a injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain and a neuregulin ERBB3 peptide), optionally in combination with chemotherapy. Within 3 weeks a significant decrease in size of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 5 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan,

MRI, PET scanning, Radionuclide imaging, bronchoscopy, etc.) or using the usual blood tests recommended for this cancer.

Example 13 - Method for treating prostate cancer

A 72 year old man is diagnosed with a stage II prostate cancer. To treat and/ or to prevent metastasis he receives a intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a GS20 linker, and an IGF-1 peptide), optionally in combination with androgen deprivative treatment. Within 10 days a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 5 weeks later no tumour is observable anymore with the usual detection tools (X-ray, ProstaScint scan, MRI, transrectal ultrasonography, CT scan, etc.) and the levels of PSA came back to normal.

Example 14 - Method for treating cervical cancer

A 60 year old woman diagnosed with cervical cancer at a limited stage is treated with surgery. To improve the effects of the treatment and to prevent metastasis she receives an intravenous injection of a polypeptide of the present invention (eg. botulinum type D neurotoxin protease, a botulinum type D neurotoxin

- 902015268754 16 Dec 2015 translocation domain, a GS20 linker, and a somatostatin-14 peptide). Within 6 weeks no re-appearance of the tumour is observed,. The treatment is repeated 3 months later and 8 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan, MRI, PET scanning, Radionuclide imaging, bronchoscopy, etc.) or using the usual blood tests recommended for this cancer.

Example 15 - Method for treating leukaemia

A 42 year old man is diagnosed with a stage II Hairy cell leukaemia cancer. To treat and/ or to prevent metastasis he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain, a GS20 linker, and a FGF-1 peptide), optionally in combination with chemotherapy. Within 10 days significant reduction in tumour burden is observed without appearance of metastasis. The treatment is repeated 1 month later and 3 weeks later no tumour is observable anymore with the usual detection tools (MRI, complete blood count, etc).

Example 16 - Method for treating small cel, lung cancer

A 56 year old man is diagnosed with a small cell lung cancer at an extensive stage. To treat and/ or to prevent metastasis elsewhere he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain and an EGF peptide), optionally in combination with chemotherapy and radiation therapy. Within 3 weeks a significant shrinkage of the tumour and a diminution in size of the metastasis is observed without appearance of new metastasis elsewhere. The treatment is repeated twice after 2 months and 5months. The patients died 11 months later, 6 months later than expected with this type of treatment and this stage of the disease.

Example 17 - Method for treating pancreatic cancer

A 48 year old woman is diagnosed with pancreatic cancer at an advanced stage. She receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C

- 91 2015268754 16 Dec 2015 neurotoxin translocation domain and an EGF peptide), optionally in combination with the chemotherapeutic agent, Gemcitabine. Within 3 weeks a significant reduction in primary tumour growth and a diminution in size of the metastases are observed without appearance of new metastasis elsewhere.

The treatment is repeated twice after 2 months and 5 months. The patient died 12 months later, 6 months later than expected with this type of treatment and this stage of the disease.

Example 18 - Method for treating metastatic bone cancer

A 71 year old man is diagnosed with a stage IV prostate cancer. To treat metastatic growth in his bone he receives an intravenous injection of a polypeptide of the present invention (eg. a botuiinum type D neurotoxin protease, a botuiinum type D neurotoxin translocation domain and a TGF-beta peptide), optionally in combination with external beam radiation plus hormone therapy. Within 4 weeks a significant shrinkage of the tumour at the metastatic sites is observed without appearance of metastasis elsewhere. The treatment is repeated 2 and 4 months later. After 6 months no detectable increase in tumour burden is observable anymore with the usual detection tools (X-ray, ProstaScint scan, MRI, CT scan, etc.).

Example 19 - Method for treating metastatic small cell lung cancer in the brain

A 65 year old man diagnosed with a small cell lung cancer at an advanced stage also presents with multiple brain metastases. To treat, a polypeptide of the present invention (eg. a botuiinum type A neurotoxin protease, a botuiinum type A neurotoxin translocation domain, and a SDF-1 peptide) is delivered into his brain by convection enhanced delivery, optionally in combination with chemotherapy and whole brain radiation. Within 4 weeks significant shrinkage of the metastatic tumour sites is observed without appearance of metastasis elsewhere. The treatment is repeated at 2 months and 8 weeks later no metastatic tumour is observable anymore with the usual detection tools (X-rays, CT scan, MRI, PET scanning, Radionuclide imaging, etc.) or using the usual blood tests recommended for this cancer.

-922015268754 16 Dec 2015

Example 20 - Method for treating bowel cancer

A 76 year old man is diagnosed with a stage II small bowel cancer. To treatment and/ or to prevent metastasis he receives a direct injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease and translocation domain and an EGF peptide), optionally in combination with chemotherapy. Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere and surgery is then realized to remove the tumour.

Example 21 - Method for treating chronic lymphocytic leukaemia

A 54 year old man is diagnosed with relapsed chronic lymphocytic leukaemia. To treat, he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type B neurotoxin protease, a botulinum type B neurotoxin translocation domain and an LIF-1 peptide), optionally in combination with chemotherapy. Within 2 weeks a significant reduction in tumour cell count in the patient’s blood is observed which remains stable for an extended period of time as determined by microscopic examination and flow cytometric analysis of the patient’s blood

Example 22 - Method for treating liver cancer

A 55 year old woman is diagnosed with a stage IV hepatic cancer. To treat, she receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain and a TGF-alpha peptide), optionally in combination with chemotherapy. Within 2 weeks a significant shrinkage of the tumour is observed on PET-CT imaging studies along with a reduction of alphafetoprotein in the blood, which was elevated prior to treatment.

Example 23 - Method for treating Hodgkin’s lymphoma

A 24 year old man is diagnosed with stage IVA Hodgkin’s lymphoma. To treat, he receives repeated intravenous injections of a polypeptide of the present invention (eg. a botulinum type B neurotoxin protease, a botulinum type B neurotoxin translocation domain and an VEGF peptide), optionally in combination with chemotherapy. Within 2 weeks a significant reduction in

- 93 2015268754 16 Dec 2015 tumour volume is observed and tumour blood flow using the usual detection tools (MRI, CT scans) and leads to complete remission within 2 months.

Example 24 - Method for treating renal cancer

A 54 year old man is diagnosed with advanced renal cell carcinoma. To treat, he receives an intravenous administration of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botuiinum type C neurotoxin translocation domain and a VEGF peptide), optionally in combination with the anti-angiogenic therapeutic, Sunitinib, a small molecule tyrosine kinase inhibitor (TKI). Within 14 days a significant shrinkage of the tumour over and above that expected with TKI alone is observed using the usual detection tools (CT, MRI scans, blood tests).

Example 25 - Method for treating skin cancer

A 31 year old woman is diagnosed with a facial melanoma. To treat, she receives direct administration of a polypeptide of the present invention (eg. a botuiinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a GS20 linker, and a VEGF peptide), optionally in combination with chemotherapy and immunotherapy. Within 21 days a significant shrinkage of the tumour over and above that expected with chemoand immunotherapy alone is observed, allowing surgical resection with clear but much reduced margins.

Example 26 - Method for treating oropharyngeal cancer

A 63 year old man is diagnosed with a stage III head and neck cancer. To treat, he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain and a TGF-alpha peptide), optionally in combination with radiotherapy and chemotherapy. Within 4 weeks a significant improvement in the patient’s ability to swallow food is apparent which is maintained longer than would be expected with the combination therapy alone.

Example 27 - Method for treating myeloma cancer

A 73 year old woman diagnosed with multiple myeloma associated osteolytic - 942015268754 16 Dec 2015 bone lesions and hypercalcemia is treated with the standard chemotherapy. To improve the effects of the treatments she receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain and an IL-6 peptide). Within 4 weeks a significant reduction in the size of bone lesions and severity of hypercalcemia is observed without appearance of new lesions elsewhere using the usual detection tools (MRI, X-ray, blood tests).

Example 28 - Method for treating soft tissue sarcoma cancer

A 51 year old woman diagnosed with a fibrosarcoma of the leg is treated with radiation therapy in an attempt to reduce tumour size prior to surgical resection. To improve the effects of the radiation treatment she receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a

GS20 linker, and a bFGF peptide). Within 14 days a significant shrinkage of the tumour over and above that expected with radiation therapy alone is observed, allowing surgical resection with clear margins.

Example 29 - Method for treating gastric cancer

A 84 year old man diagnosed with advanced gastric cancer and unable to undergo surgical resection is treated with radiation therapy to relieve tumour associated blockage. To improve the effects of the treatment he receives multiple direct injections of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain, a GS20 linker, and a GRP peptide). Within 5 days a significant shrinkage of the tumour is observed with the usual detection tools (gastroscopic examination and CT scan). The treatment is repeated 1 month later and 4 months later tumour blockage has not recurred.

Example 30 - Method for treating testicular cancer

A 32 year old man is diagnosed with a stage I seminoma cancer. To treat and/ or to prevent recurrence he receives a intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a GS20 linker, and a VEGF peptide),

- 95 2015268754 16 Dec 2015 optionally in combination with chemotherapy. Within 14 days a significant shrinkage of the tumour is observed. The treatment is repeated 1 month later and 6 weeks later no tumour is observable anymore with the usual detection tools (blood tests and CT scans).

Example 31 - Method for treating uterine cancer

A 76 year old woman diagnosed with a stage IIA endometrial cancer is treated with usual chemotherapy and radiotherapy. To improve the effects of the treatment and to prevent metastasis she receives a direct injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease and translocation domain and an EGF peptide). Within 6 weeks a significant shrinkage of the tumour is observed on direct visualization of the uterine cavity by hysteroscopy without appearance of metastasis elsewhere.

Example 32 - Method for treating Karposi sarcoma

A 48 year old man is diagnosed with acquired immunity deficiency syndrome (AIDS) and presents with multiple Karposi sarcoma lesions. To treat, he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain and an IL-6 peptide), optionally in combination with interferon alpha. Within 3 weeks a significant reduction in lesion size is observed without appearance of new lesions elsewhere. The treatment is repeated twice after 1 month and 3 months which effectively stops the progression of the Kaposi sarcoma.

Example 33 - Method for treating primary brain cancer

A 45 year old woman is diagnosed with glioblastoma. To treat and/ or to prevent further metastasis she receives an intracranial application of a polypeptide of the present invention (eg. a botulinum type C neurotoxin protease, a botulinum type C neurotoxin translocation domain and a VEGF peptide), optionally in combination with chemotherapy. Within 4 weeks a significant decrease in the size of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 1 month later and 4 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan, etc.).

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Example 34 - Method for treating rectal cancer

A 77 year old man diagnosed with a stage II rectal cancer is treated with a polypeptide of the present invention (eg. botulinum type A neurotoxin protease and an anti-EGFR antibody F(ab)’₂ fragment), optionally incombination with radiotherapy and surgery. He receives localised injections of polypeptide (eg. 3 days in advance of a standard regiment of fractionated radiation) and within 2 weeks a significant shrinkage of the tumour is observed which enables complete surgical resection. 12 months later no tumour recurrence is observable with the usual detection tools (colonoscopy, CT scan, PET scan, etc.) and the level of carcinoembryonic antigen (CEA) returned to the normal.

Example 35 - Assessment of proliferation changes, inhibition of cellular secretion and concomitant SNARE cleavage after treatment of in vitro cultured renal cancer cell lines with a polypeptide of the present invention.

Methods:

Proliferation analysis:

An appropriate volume containing 1000 cells of a suitable renal cancer cell line, for example A498, ACHN or 786-0, are seeded into the wells of a 96-well cell culture plate in a suitable growth medium supplemented with 10% Foetal Bovine Serum and incubated at 37°C in a humidified atmosphere with 5% CO₂. Cells are allowed to adhere overnight after which treatment with an

EGF-liganded LHA molecule, such as an LHA molecule with a C-terminal presented EGF ligand, is initiated. After 24 hours, the treatment media are removed, cell monolayers are washed to remove traces of LHA-EGF and fresh medium applied to cells. After a further 24 hours a conventional colorimetric proliferation assay (based on the determination of the cleavage of the tetrazolium salt WST-1 to formazan by cellular enzymes) is performed. Specifically, WST-1 is added to the culture medium for 4 hours after which the optical density at 440 nm is determined for each treatment. Figures 50-52 demonstrate inhibition of in vitro proliferation from a renal cell carcinoma line

- 972015268754 16 Dec 2015 by an EGF-LHA fusion.

Cellular Secretion analysis:

10,000 cells of a suitable renal cancer cell line (for example 786-0, A498 or

ACHN) are seeded in the wells of a 24 well plate in an appropriate culture medium containing 10% foetal bovine serum. Plates are incubated overnight in an incubator at 37°C in a humidified atmosphere with 5% CO₂to allow cells to adhere. Cell cultures are then treated with an appropriate polypeptide of the present invention which targets a specific receptor on the cells of interest (for the cell lines detailed above an appropriate molecule would be an EGFliganded LHA as these 3 cell lines all express EGF receptors, Figure 48). After 24 hours, the treatment medium is removed, cell monolayers refed with fresh culture medium and 24-48 hours later samples of the culture medium are removed to fresh microfuge tubes which are subsequently spun at 1500 rpm for 5 minutes to remove floating cells. The resultant supernatants are removed to fresh tubes and aliquots then used for the quantification of specific analytes (for example VEGF, TNF-alpha) by enzyme-linked immunosorbent assay or ELISA (Figure 53). An example of analysis of inhibition of a secretion (GRP) from a small cell lung carcinoma cell line is provided in Figure 54.

Western blot analysis to detect SNARE cleavage due to cellular uptake of polypeptides of the present invention:

Cell cultures are treated with polypeptides as detailed above for the analysis of cellular secretions. After removal of culture medium for subsequent ELISA analysis, cell monolayers in each well are washed three times with phosphatebuffered saline and protein extracts prepared by cellular lysis in standard Laemmli sample buffer. Cellular protein are separated on 12% SDSpolyacrylamide gels and electrophoretically transferred to a nitrocellulose membrane. After blocking of the membrane, primary antisera are used to probe for the SNARE protein of interest and detection of full length and/or cleaved forms is enabled by a peroxidase-conjugated anti-species IgG. In the case of treatment of the renal cell line 786-0 with an LHA-EGF molecule, cleavage of the SNARE SNAP-25 is observed (see Figure 49).

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SEQ ID NOs

1. DNA sequence of LH_N/A ggatccATGGAGTTCGTTAACAAACAGTTCAACTATAAAGACCCAGTTAACGGTGTTGACATTGCTTAC

ATCAAAATCCCGAACGCTGGCCAGATGCAGCCGGTAAAGGCATTCAAAATCCACAACAAAATCTGGGTT

ATCCCGGAACGTGATACCTTTACTAACCCGGAAGAAGGTGACCTGAACCCGCCACCGGAAGCGAAACAG

GTGCCGGTATCTTACTATGACTCCACCTACCTGTCTACCGATAACGAAAAGGACAACTACCTGAAAGGT GTTACTAAACTGTTCGAGCGTATTTACTCCACCGACCTGGGCCGTATGCTGCTGACTAGCATCGTTCGC GGTATCCCGTTCTGGGGCGGTTCTACCATCGATACCGAACTGAAAGTAATCGACACTAACTGCATCAAC gttattcagccggacggttcctatcgttccgaagaactgaacctggtgatcatcggcccgtctgctgat ATCATCCAGTTCGAGTGTCTGAGCTTTGGTCACGAAGTTCTGAACCTCACCCGTAACGGCTACGGTTCC

ACTCAGTACATCCGTTTCTCTCCGGACTTCACCTTCGGTTTTGAAGAATCCCTGGAAGTAGACACGAAC CCACTGCTGGGCGCTGGTAAATTCGCAACTGATCCTGCGGTTACCCTGGCTCACGAACTGATTCATGCA GGCCACCGCCTGTACGGTATCGCCATCAATCCGAACCGTGTCTTCAAAGTTAACACCAACGCGTATTAC GAGATGTCCGGTCTGGAAGTTAGCTTCGAAGAACTGCGTACTTTTGGCGGTCACGACGCTAAATTCATC GACTCTCTGCAAGAAAACGAGTTCCGTCTGTACTACTATAACAAGTTCAAAGATATCGCATCCACCCTG

AACAAAGCGAAATCCATCGTGGGTACCACTGCTTCTCTCCAGTACATGAAGAACGTTTTTAAAGAAAAA TACCTGCTCAGCGAAGACACCTCCGGCAAATTCTCTGTAGACAAGTTGAAATTCGATAAACTTTACAAA ATGCTGACTGAAATTTACACCGAAGACAACTTCGTTAAGTTCTTTAAAGTTCTGAACCGCAAAACCTAT CTGAACTTCGACAAGGCAGTATTCAAAATCAACATCGTGCCGAAAGTTAACTACACTATCTACGATGGT TTCAACCTGCGTAACACCAACCTGGCTGCTAATTTTAACGGCCAGAACACGGAAATCAACAACATGAAC

TTCACAAAACTGAAAAACTTCACTGGTCTGTTCGAGTTTTACAAGCTGCTGTGCGTCGACGGCATCATT ACCTCCAAAACTAAATCTGACGATGACGATAAAAACAAAGCGCTGAACCTGCAGTGTATCAAGGTTAAC AACTGGGATTTATTCTTCAGCCCGAGTGAAGACAACTTCACCAACGACCTGAACAAAGGTGAAGAAATC ACCTCAGATACTAACATCGAAGCAGCCGAAGAAAACATCTCGCTGGACCTGATCCAGCAGTACTACCTG ACCTTTAATTTCGACAACGAGCCGGAAAACATTTCTATCGAAAACCTGAGCTCTGATATCATCGGCCAG

CTGGAACTGATGCCGAACATCGAACGTTTCCCAAACGGTAAAAAGTACGAGCTGGACAAATATACCATG TTCCACTACCTGCGCGCGCAGGAATTTGAACACGGCAAATCCCGTATCGCACTGACTAACTCCGTTAAC GAAGCTCTGCTCAACCCGTCCCGTGTATACACCTTCTTCTCTAGCGACTACGTGAAAAAGGTCAACAAA GCGACTGAAGCTGCAATGTTCTTGGGTTGGGTTGAACAGCTTGTTTATGATTTTACCGACGAGACGTCC GAAGTATCTACTACCGACAAAATTGCGGATATCACTATCATCATCCCGTACATCGGTCCGGCTCTGAAC

ATTGGCAACATGCTGTACAAAGACGACTTCGTTGGCGCACTGATCTTCTCCGGTGCGGTGATCCTGCTG GAGTTCATCCCGGAAATCGCCATCCCGGTACTGGGCACCTTTGCTCTGGTTTCTTACATTGCAAACAAG GTTCTGACTGTACAAACCATCGACAACGCGCTGAGCAAACGTAACGAAAAATGGGATGAAGTTTACAAA TATATCGTGACCAACTGGCTGGCTAAGGTTAATACTCAGATCGACCTCATCCGCAAAAAAATGAAAGAA GCACTGGAAAACCAGGCGGAAGCTACCAAGGCAATCATTAACTACCAGTACAACCAGTACACCGAGGAA

GAAAAAAACAACATCAACTTCAACATCGACGATCTGTCCTCTAAACTGAACGAATCCATCAACAAAGCT ATGATCAACATCAACAAGTTCCTGAACCAGTGCTCTGTAAGCTATCTGATGAACTCCATGATCCCGTAC GGTGTTAAACGTCTGGAGGACTTCGATGCGTCTCTGAAAGACGCCCTGCTGAAATACATTTACGACAAC CGTGGCACTCTGATCGGTCAGGTTGATCGTCTGAAGGACAAAGTGAACAATACCTTATCGACCGACATC CCTTTTCAGCTCAGTAAATATGTCGATAACCAACGCCTTTTGTCCACTctagaataatgaaagctt

2. DNA sequence of LH_N/B

GGATCCATGCCGGTTACCATCAACAACTTCAACTACAACGACCCGATCGACAACAACAACATCATTATG

ATGGAACCGCCGTTCGCACGTGGTACCGGACGTTACTACAAGGCTTTTAAGATCACGGACCGTATCTGG

ATCATCCCGGAACGTTACACCTTCGGTTACAAACCTGAGGACTTCAACAAGAGTAGCGGGATTTTCAAT CGTGACGTCTGCGAGTACTATGATCCAGATTATCTGAATACCAACGATAAGAAGAACATATTCCTTCAG ACTATGATTAAACTCTTCAACCGTATCAAAAGCAAACCGCTCGGTGAAAAACTCCTCGAAATGATTATC AACGGTATCCCGTACCTCGGTGACCGTCGTGTCCCGCTTGAAGAGTTCAACACCAACATCGCAAGCGTC ACCGTCAACAAACTCATCAGCAACCCAGGTGAAGTCGAACGTAAAAAAGGTATCTTCGCAAACCTCATC

ATCTTCGGTCCGGGTCCGGTCCTCAACGAAAACGAAACCATCGACATCGGTATCCAGAACCACTTCGCA AGCCGTGAAGGTTTCGGTGGTATCATGCAGATGAAATTCTGCCCGGAATACGTCAGTGTCTTCAACAAC GTCCAGGAAAACAAAGGTGCAAGCATCTTCAACCGTCGTGGTTACTTCAGCGACCCGGCACTCATCCTC ATGCATGAACTCATCCACGTCCTCCACGGTCTCTACGGTATCAAAGTTGACGACCTCCCGATCGTCCCG AACGAGAAGAAATTCTTCATGCAGAGCACCGACGCAATCCAGGCTGAGGAACTCTACACCTTCGGTGGC

-99Ο

CM

2015268754 16 Dec

CAAGACCCAAGTATCATAACCCCGTCCAGCGACAAAAGCATCTACGACAAAGTCCTCCAGAACTTCAGG

GGTATCGTGGACAGACTCAACAAAGTCCTCGTCTGCATCAGCGACCCGAACATCAATATCAACATATAC

AAGAACAAGTTCAAAGACAAGTACAAATTCGTCGAGGACAGCGAAGGCAAATACAGCATCGACGTAGAA

AGTTTCGACAAGCTCTACAAAAGCCTCATGTTCGGTTTCACCGAAACCAACATCGCCGAGAACTACAAG

ATCAAGACAAGGGCAAGTTACTTCAGCGACAGCCTCCCGCCTGTCAAAATCAAGAACCTCTTAGACAAC GAGAT T TACACAAT TGAAGAGGGC T T CAACAT CAGTGACAAAGACATGGAGAAGGAATACAGAGGT CAG AACAAGGCTATCAACAAACAGGCATACGAGGAGATCAGCAAAGAACACCTCGCAGTCTACAAGATCCAG ATGTGCGTCGACGGCATCATTACCTCCAAAACTAAATCTGACGATGACGATAAAAACAAAGCGCTGAAC CTGCAGTGCATCGACGTTGACAACGAAGACCTGTTCTTCATCGCTGACAAAAACAGCTTCAGTGACGAC

CTGAGCAAAAACGAACGTATCGAATACAACACCCAGAGCAACTACATCGAAAACGACTTCCCGATCAAC GAACTGATCCTGGACACCGACCTGATAAGTAAAATCGAACTGCCGAGCGAAAACACCGAAAGTCTGACC GACTTCAACGTTGACGTTCCGGTTTACGAAAAACAGCCGGCTATCAAGAAAATCTTCACCGACGAAAAC ACCATCTTCCAGTACCTGTACAGCCAGACCTTCCCGCTGGACATCCGTGACATCAGTCTGACCAGCAGT TTCGACGACGCTCTGCTGTTCAGCAACAAAGTTTACAGTTTCTTCAGCATGGACTACATCAAAACCGCT

AACAAAGTTGTTGAAGCAGGGCTGTTCGCTGGTTGGGTTAAACAGATCGTTAACGACTTCGTTATCGAA GCTAACAAAAGCAACACTATGGACAAAATCGCTGACATCAGTCTGATCGTTCCGTACATCGGTCTGGCT CTGAACGTTGGTAACGAAACCGCTAAAGGTAACTTTGAAAACGCTTTCGAGATCGCTGGTGCAAGCATC CTGCTGGAGTTCATCCCGGAACTGCTGATCCCGGTTGTTGGTGCTTTCCTGCTGGAAAGTTACATCGAC AACAAAAACAAGATCATCAAAACCATCGACAACGCTCTGACCAAACGTAACGAAAAATGGAGTGATATG

TACGGTCTGATCGTTGCTCAGTGGCTGAGCACCGTCAACACCCAGTTCTACACCATCAAAGAAGGTATG TACAAAGCTCTGAACTACCAGGCTCAGGCTCTGGAAGAGATCATCAAATACCGTTACAACATCTACAGT GAGAAGGAAAAGAGTAACATCAACATCGACTTCAACGACATCAACAGCAAACTGAACGAAGGTATCAAC CAGGCTATCGACAACATCAACAACTTCATCAACGGTTGCAGTGTTAGCTACCTGATGAAGAAGATGATC CCGCTGGCTGTTGAAAAACTGCTGGACTTCGACAACACCCTGAAAAAGAACCTGCTGAACTACATCGAC

GAAAACAAGCTGTACCTGATCGGTAGTGCTGAATACGAAAAAAGTAAAGTGAACAAATACCTGAAGACC ATCATGCCGTTCGACCTGAGTATCTACACCAACGACACCATCCTGATCGAAATGTTCAACAAATACAAC TCtctagaataatgaaagctt

3. DNA sequence of LH_N/C ggatccATGCCGATCACCATCAACAACTTCAACTACAGCGATCCGGTGGATAACAAAAACATCCTGTAC

CTGGATACCCATCTGAATACCCTGGCGAACGAACCGGAAAAAGCGTTTCGTATCACCGGCAACATTTGG

GTTATTCCGGATCGTTTTAGCCGTAACAGCAACCCGAATCTGAATAAACCGCCGCGTGTTACCAGCCCG

AAAAGCGGTTATTACGATCCGAACTATCTGAGCACCGATAGCGATAAAGATACCTTCCTGAAAGAAATC

ATCAAACTGTTCAAACGCATCAACAGCCGTGAAATTGGCGAAGAACTGATCTATCGCCTGAGCACCGAT ATTCCGTTTCCGGGCAACAACAACACCCCGATCAACACCTTTGATTTCGATGTGGATTTCAACAGCGTT GATGTTAAAACCCGCCAGGGTAACAATTGGGTGAAAACCGGCAGCATTAACCCGAGCGTGATTATTACC GGTCCGCGCGAAAACATTATTGATCCGGAAACCAGCACCTTTAAACTGACCAACAACACCTTTGCGGCG CAGGAAGGTTTTGGCGCGCTGAGCATTATTAGCATTAGCCCGCGCTTTATGCTGACCTATAGCAACGCG

ACCAACGATGTTGGTGAAGGCCGTTTCAGCAAAAGCGAATTTTGCATGGACCCGATCCTGATCCTGATG CATGAACTGAACCATGCGATGCATAACCTGTATGGCATCGCGATTCCGAACGATCAGACCATTAGCAGC GTGACCAGCAACATCTTTTACAGCCAGTACAACGTGAAACTGGAATATGCGGAAATCTATGCGTTTGGC GGTCCGACCATTGATCTGATTCCGAAAAGCGCGCGCAAATACTTCGAAGAAAAAGCGCTGGATTACTAT CGCAGCATTGCGAAACGTCTGAACAGCATTACCACCGCGAATCCGAGCAGCTTCAACAAATATATCGGC

GAATATAAACAGAAACTGATCCGCAAATATCGCTTTGTGGTGGAAAGCAGCGGCGAAGTTACCGTTAAC CGCAATAAATTCGTGGAACTGTACAACGAACTGACCCAGATCTTCACCGAATTTAACTATGCGAAAATC TATAACGTGCAGAACCGTAAAATCTACCTGAGCAACGTGTATACCCCGGTGACCGCGAATATTCTGGAT GATAACGTGTACGATATCCAGAACGGCTTTAACATCCCGAAAAGCAACCTGAACGTTCTGTTTATGGGC CAGAACCTGAGCCGTAATCCGGCGCTGCGTAAAGTGAACCCGGAAAACATGCTGTACCTGTTCACCAAA

TTTTGCGTCGACGCGATTGATGGTCGTAGCCTGTACAACAAAACCCTGCAGTGTCGTGAACTGCTGGTG AAAAACACCGATCTGCCGTTTATTGGCGATATCAGCGATGTGAAAACCGATATCTTCCTGCGCAAAGAT ATCAACGAAGAAACCGAAGTGATCTACTACCCGGATAACGTGAGCGTTGATCAGGTGATCCTGAGCAAA AACACCAGCGAACATGGTCAGCTGGATCTGCTGTATCCGAGCATTGATAGCGAAAGCGAAATTCTGCCG GGCGAAAACCAGGTGTTTTACGATAACCGTACCCAGAACGTGGATTACCTGAACAGCTATTACTACCTG

GAAAGCCAGAAACTGAGCGATAACGTGGAAGATTTTACCTTTACCCGCAGCATTGAAGAAGCGCTGGAT AACAGCGCGAAAGTTTACACCTATTTTCCGACCCTGGCGAACAAAGTTAATGCGGGTGTTCAGGGCGGT CTGTTTCTGATGTGGGCGAACGATGTGGTGGAAGATTTCACCACCAACATCCTGCGTAAAGATACCCTG GATAAAATCAGCGATGTTAGCGCGATTATTCCGTATATTGGTCCGGCGCTGAACATTAGCAATAGCGTG CGTCGTGGCAATTTTACCGAAGCGTTTGCGGTTACCGGTGTGACCATTCTGCTGGAAGCGTTTCCGGAA

TTTACCATTCCGGCGCTGGGTGCGTTTGTGATCTATAGCAAAGTGCAGGAACGCAACGAAATCATCAAA ACCATCGATAACTGCCTGGAACAGCGTATTAAACGCTGGAAAGATAGCTATGAATGGATGATGGGCACC

- 1002015268754 16 Dec 2015

TGGCTGAGCCGTATTATCACCCAGTTCAACAACATCAGCTACCAGATGTACGATAGCCTGAACTATCAG

GCGGGTGCGATTAAAGCGAAAATCGATCTGGAATACAAAAAATACAGCGGCAGCGATAAAGAAAACATC

AAAAGCCAGGTTGAAAACCTGAAAAACAGCCTGGATGTGAAAATTAGCGAAGCGATGAATAACATCAAC

AAATTCATCCGCGAATGCAGCGTGACCTACCTGTTCAAAAACATGCTGCCGAAAGTGATCGATGAACTG

AACGAATTTGATCGCAACACCAAAGCGAAACTGATCAACCTGATCGATAGCCACAACATTATTCTGGTG GGCGAAGTGGATAAACTGAAAGCGAAAGTTAACAACAGCTTCCAGAACACCATCCCGTTTAACATCTTC AGCTATACCAACAACAGCCTGCTGAAAGATATCATCAACGAATACTTCAAtctagaataatgaaagctt

4. DNA sequence of LH_N/D ggatccATGACGTGGCCAGTTAAGGATTTCAACTACTCAGATCCTGTAAATGACAACGATATTCTGTAC

CTTCGCATTCCACAAAATAAACTGATCACCACACCAGTCAAAGCATTCATGATTACTCAAAACATTTGG

GTCATTCCAGAACGCTTTTCTAGTGACACAAATCCGAGTTTATCTAAACCTCCGCGTCCGACGTCCAAA

TATCAGAGCTATTACGATCCCTCATATCTCAGTACGGACGAACAAAAAGATACTTTCCTTAAAGGTATC

ATTAAACTGTTTAAGCGTATTAATGAGCGCGATATCGGGAAAAAGTTGATTAATTATCTTGTTGTGGGT TCCCCGTTCATGGGCGATAGCTCTACCCCCGAAGACACTTTTGATTTTACCCGTCATACGACAAACATC GCGGTAGAGAAGTTTGAGAACGGATCGTGGAAAGTCACAAACATCATTACACCTAGCGTCTTAATTTTT GGTCCGCTGCCAAACATCTTAGATTATACAGCCAGCCTGACTTTGCAGGGGCAACAGTCGAATCCGAGT TTCGAAGGTTTTGGTACCCTGAGCATTCTGAAAGTTGCCCCGGAATTTCTGCTCACTTTTTCAGATGTC

ACCAGCAACCAGAGCTCAGCAGTATTAGGAAAGTCAATTTTTTGCATGGACCCGGTTATTGCACTGATG CACGAACTGACGCACTCTCTGCATCAACTGTATGGGATCAACATCCCCAGTGACAAACGTATTCGTCCC CAGGTGTCTGAAGGATTTTTCTCACAGGATGGGCCGAACGTCCAGTTCGAAGAGTTGTATACTTTCGGA GGCCTGGACGTAGAGATCATTCCCCAGATTGAGCGCAGTCAGCTGCGTGAGAAGGCATTGGGCCATTAT AAGGATATTGCAAAACGCCTGAATAACATTAACAAAACGATTCCATCTTCGTGGATCTCGAATATTGAT

AAATATAAGAAAATTTTTAGCGAGAAATATAATTTTGATAAAGATAATACAGGTAACTTTGTGGTTAAC ATTGACAAATTCAACTCCCTTTACAGTGATTTGACGAATGTAATGAGCGAAGTTGTGTATAGTTCCCAA TACAACGTTAAGAATCGTACCCATTACTTCTCTCGTCACTACCTGCCGGTTTTCGCGAACATCCTTGAC GATAATATTTACACTATTCGTGACGGCTTTAACTTGACCAACAAGGGCTTCAATATTGAAAATTCAGGC CAGAACATTGAACGCAACCCGGCCTTGCAGAAACTGTCGAGTGAATCCGTGGTTGACCTGTTTACCAAA

GTCTGCGTCGACAAAAGCGAAGAGAAGCTGTACGATGACGATGACAAAGATCGTTGGGGATCGTCCCTG CAGTGTATTAAAGTGAAAAACAATCGGCTGCCTTATGTAGCAGATAAAGATAGCATTAGTCAGGAGATT TTCGAAAATAAAATTATCACTGACGAAACCAATGTTCAGAATTATTCAGATAAATTTTCACTGGACGAA AGCATCTTAGATGGCCAAGTTCCGATTAACCCGGAAATTGTTGATCCGTTACTGCCGAACGTGAATATG GAACCGTTAAACCTCCCTGGCGAAGAGATCGTATTTTATGATGACATTACGAAATATGTGGACTACCTT

AATTCTTATTACTATTTGGAAAGCCAGAAACTGTCCAATAACGTGGAAAACATTACTCTGACCACAAGC GTGGAAGAGGCTTTAGGCTACTCAAATAAGATTTATACCTTCCTCCCGTCGCTGGCGGAAAAAGTAAAT AAAGGTGTGCAGGCTGGTCTGTTCCTCAACTGGGCGAATGAAGTTGTCGAAGACTTTACCACGAATATT ATGAAAAAGGATACCCTGGATAAAATCTCCGACGTCTCGGTTATTATCCCATATATTGGCCCTGCGTTA AATATCGGTAATAGTGCGCTGCGGGGGAATTTTAACCAGGCCTTTGCTACCGCGGGCGTCGCGTTCCTC

CTGGAGGGCTTTCCTGAATTTACTATCCCGGCGCTCGGTGTTTTTACATTTTACTCTTCCATCCAGGAG CGTGAGAAAATTATCAAAACCATCGAAAACTGCCTGGAGCAGCGGGTGAAACGCTGGAAAGATTCTTAT CAATGGATGGTGTCAAACTGGTTATCTCGCATCACGACCCAATTCAACCATATTAATTACCAGATGTAT GATAGTCTGTCGTACCAAGCTGACGCCATTAAAGCCAAAATTGATCTGGAATATAAAAAGTACTCTGGT AGCGATAAGGAGAACATCAAAAGCCAGGTGGAGAACCTTAAGAATAGTCTGGATGTGAAAATCTCTGAA

GCTATGAATAACATTAACAAATTCATTCGTGAATGTTCGGTGACGTACCTGTTCAAGAATATGCTGCCA AAAGTTATTGATGAACTGAATAAATTTGATCTGCGTACCAAAACCGAACTTATCAACCTCATCGACTCC CACAACATTATCCTTGTGGGCGAAGTGGATCGTCTGAAGGCCAAAGTAAACGAGAGCTTTGAAAATACG ATGCCGTTTAATATTTTTTCATATACCAATAACTCCTTGCTGAAAGATATCATCAATGAATATTTCAAT ctagattaataagctt

5. DNA sequence of the CT-CST28 linker

GGATCCGTCGACCTGCAGGGTCTAGAAGGCGGTGGCGGTAGCGGCGGTGGCGGTAGCGGCGGTGGCGGT

AGCGGCGGTGGCGGTAGCGCACTAGTGCAGGAAAGACCTCCATTACAACAACCTCCACATCGCGATAAG

AAACCATGTAAGAATTTCTTTTGGAAAACATTTAGCAGTTGCAAATGATAAAAGCTT

6. DNA sequence of the LHD-CT-CST28 fusion

GGATCCATGACGTGGCCAGTTAAGGATTTCAACTACTCAGATCCTGTAAATGACAACGATATTCTGTAC

CTTCGCATTCCACAAAATAAACTGATCACCACACCAGTCAAAGCATTCATGATTACTCAAAACATTTGG GTCATTCCAGAACGCTTTTCTAGTGACACAAATCCGAGTTTATCTAAACCTCCGCGTCCGACGTCCAAA

- 101 2015268754 16 Dec 2015

TATCAGAGCTATTACGATCCCTCATATCTCAGTACGGACGAACAAAAAGATACTTTCCTTAAAGGTATC

ATTAAACTGTTTAAGCGTATTAATGAGCGCGATATCGGGAAAAAGTTGATTAATTATCTTGTTGTGGGT

TCCCCGTTCATGGGCGATAGCTCTACCCCCGAAGACACTTTTGATTTTACCCGTCATACGACAAACATC

GCGGTAGAGAAGTTTGAGAACGGATCGTGGAAAGTCACAAACATCATTACACCTAGCGTCTTAATTTTT

GGTCCGCTGCCAAACATCTTAGATTATACAGCCAGCCTGACTTTGCAGGGGCAACAGTCGAATCCGAGT TTCGAAGGTTTTGGTACCCTGAGCATTCTGAAAGTTGCCCCGGAATTTCTGCTCACTTTTTCAGATGTC ACCAGCAACCAGAGCTCAGCAGTATTAGGAAAGTCAATTTTTTGCATGGACCCGGTTATTGCACTGATG CACGAACTGACGCACTCTCTGCATCAACTGTATGGGATCAACATCCCCAGTGACAAACGTATTCGTCCC CAGGTGTCTGAAGGATTTTTCTCACAGGATGGGCCGAACGTCCAGTTCGAAGAGTTGTATACTTTCGGA

GGCCTGGACGTAGAGATCATTCCCCAGATTGAGCGCAGTCAGCTGCGTGAGAAGGCATTGGGCCATTAT AAGGATATTGCAAAACGCCTGAATAACATTAACAAAACGATTCCATCTTCGTGGATCTCGAATATTGAT AAATATAAGAAAATTTTTAGCGAGAAATATAATTTTGATAAAGATAATACAGGTAACTTTGTGGTTAAC ATTGACAAATTCAACTCCCTTTACAGTGATTTGACGAATGTAATGAGCGAAGTTGTGTATAGTTCCCAA TACAACGTTAAGAATCGTACCCATTACTTCTCTCGTCACTACCTGCCGGTTTTCGCGAACATCCTTGAC

GATAATATTTACACTATTCGTGACGGCTTTAACTTGACCAACAAGGGCTTCAATATTGAAAATTCAGGC CAGAACATTGAACGCAACCCGGCCTTGCAGAAACTGTCGAGTGAATCCGTGGTTGACCTGTTTACCAAA GTCTGCGTCGACAAAAGCGAAGAGAAGCTGTACGATGACGATGACAAAGATCGTTGGGGATCGTCCCTG CAGTGTATTAAAGTGAAAAACAATCGGCTGCCTTATGTAGCAGATAAAGATAGCATTAGTCAGGAGATT TTCGAAAATAAAATTATCACTGACGAAACCAATGTTCAGAATTATTCAGATAAATTTTCACTGGACGAA

AGCATCTTAGATGGCCAAGTTCCGATTAACCCGGAAATTGTTGATCCGTTACTGCCGAACGTGAATATG GAACCGTTAAACCTCCCTGGCGAAGAGATCGTATTTTATGATGACATTACGAAATATGTGGACTACCTT AATTCTTATTACTATTTGGAAAGCCAGAAACTGTCCAATAACGTGGAAAACATTACTCTGACCACAAGC GTGGAAGAGGCTTTAGGCTACTCAAATAAGATTTATACCTTCCTCCCGTCGCTGGCGGAAAAAGTAAAT AAAGGTGTGCAGGCTGGTCTGTTCCTCAACTGGGCGAATGAAGTTGTCGAAGACTTTACCACGAATATT

ATGAAAAAGGATACCCTGGATAAAATCTCCGACGTCTCGGTTATTATCCCATATATTGGCCCTGCGTTA AATATCGGTAATAGTGCGCTGCGGGGGAATTTTAACCAGGCCTTTGCTACCGCGGGCGTCGCGTTCCTC CTGGAGGGCTTTCCTGAATTTACTATCCCGGCGCTCGGTGTTTTTACATTTTACTCTTCCATCCAGGAG CGTGAGAAAATTATCAAAACCATCGAAAACTGCCTGGAGCAGCGGGTGAAACGCTGGAAAGATTCTTAT CAATGGATGGTGTCAAACTGGTTATCTCGCATCACGACCCAATTCAACCATATTAATTACCAGATGTAT

GATAGTCTGTCGTACCAAGCTGACGCCATTAAAGCCAAAATTGATCTGGAATATAAAAAGTACTCTGGT AGCGATAAGGAGAACATCAAAAGCCAGGTGGAGAACCTTAAGAATAGTCTGGATGTGAAAATCTCTGAA GCTATGAATAACATTAACAAATTCATTCGTGAATGTTCGGTGACGTACCTGTTCAAGAATATGCTGCCA AAAGTTATTGATGAACTGAATAAATTTGATCTGCGTACCAAAACCGAACTTATCAACCTCATCGACTCC CACAACATTATCCTTGTGGGCGAAGTGGATCGTCTGAAGGCCAAAGTAAACGAGAGCTTTGAAAATACG

ATGCCGTTTAATATTTTTTCATATACCAATAACTCCTTGCTGAAAGATATCATCAATGAATATTTCAAT CTAGAAGGCGGTGGCGGTAGCGGCGGTGGCGGTAGCGGCGGTGGCGGTAGCGCACTAGTGCAGGAAAGA CCTCCATTACAACAACCTCCACATCGCGATAAGAAACCATGTAAGAATTTCTTTTGGAAAACATTTAGC AGTTGCAAAtaataagctt

7. Protein sequence of the LHD-CT-CST28 fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS

YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE

KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN

QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VE 11PQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNI DKYKKI FSEKYNFDKDNTGNFWNI DK FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLS SE SWDLFTKVCVDKSEEKLYDDDDKDRWGS SLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY

YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVQERPPL

QQPPHRDKKPCKNFFWKTFSSCK

8. DNA sequence of the CP-EGF linker ggatccGTCGACaacaacaataacaacaacaataacaacaacgacgatgacgataaaAATTCAGATAGC

GAATGTCCACTTAGTCACGACGGGTACTGTTTGCATGATGGTGTGTGTATGTATATAGAAGCACTAGAC AAATACGCTTGCAATTGCGTAGTTGGCTATATAGGAGAGCGATGCCAATATAGAGATCTGAAGTGGTGG

- 1022015268754 16 Dec 2015

GAGTTAAGGGCAgaagcggcagccaaagaagcagccgctaaggcgctgcagagtctagaataataagct t

9. DNA sequence of the LHA-CP-EGF fusion ggatccATGGAGTTCGTTAACAAACAGTTCAACTATAAAGACCCAGTTAACGGTGTTGACATTGCTTAC

ATCAAAATCCCGAACGCTGGCCAGATGCAGCCGGTAAAGGCATTCAAAATCCACAACAAAATCTGGGTT

ATCCCGGAACGTGATACCTTTACTAACCCGGAAGAAGGTGACCTGAACCCGCCACCGGAAGCGAAACAG

GTGCCGGTATCTTACTATGACTCCACCTACCTGTCTACCGATAACGAAAAGGACAACTACCTGAAAGGT

GTTACTAAACTGTTCGAGCGTATTTACTCCACCGACCTGGGCCGTATGCTGCTGACTAGCATCGTTCGC GGTATCCCGTTCTGGGGCGGTTCTACCATCGATACCGAACTGAAAGTAATCGACACTAACTGCATCAAC GTTATTCAGCCGGACGGTTCCTATCGTTCCGAAGAACTGAACCTGGTGATCATCGGCCCGTCTGCTGAT ATCATCCAGTTCGAGTGTCTGAGCTTTGGTCACGAAGTTCTGAACCTCACCCGTAACGGCTACGGTTCC ACTCAGTACATCCGTTTGTCTCCGGACTTCACCTTCGGTTTTGAAGAATCCCTGGAAGTAGACACGAAC

CCACTGCTGGGCGCTGGTAAATTCGCAACTGATCCTGCGGTTACCCTGGCTCACGAACTGATTCATGCA GGCCACCGCCTGTACGGTATCGCCATCAATCCGAACCGTGTCTTCAAAGTTAACACCAACGCGTATTAC GAGATGTCCGGTCTGGAAGTTAGCTTCGAAGAACTGCGTACTTTTGGCGGTCACGACGCTAAATTCATC GACTCTCTGCAAGAAAACGAGTTCCGTCTGTACTACTATAACAAGTTCAAAGATATCGCATCCACCCTG AACAAAGCGAAATCCATCGTGGGTACCACTGCTTCTCTCCAGTACATGAAGAACGTTTTTAAAGAAAAA

TACCTGCTCAGCGAAGACACCTCCGGCAAATTCTCTGTAGACAAGTTGAAATTCGATAAACTTTACAAA ATGCTGACTGAAATTTACACCGAAGACAACTTCGTTAAGTTCTTTAAAGTTCTGAACCGCAAAACCTAT CTGAACTTCGACAAGGCAGTATTCAAAATCAACATCGTGCCGAAAGTTAACTACACTATCTACGATGGT TTCAACCTGCGTAACACCAACCTGGCTGCTAATTTTAACGGCCAGAACACGGAAATCAACAACATGAAC TTCACAAAACTGAAAAACTTCACTGGTCTGTTCGAGTTTTACAAGCTGCTGTGCGTCGACaacaacaat aacaacaacaataacaacaacgacgatgacgataaaAATTCAGATAGCGAATGTCCACTTAGTCACGAC GGGTACTGTTTGCATGATGGTGTGTGTATGTATATAGAAGCACTAGACAAATACGCTTGCAATTGCGTA GTTGGCTATATAGGAGAGCGATGCCAATATAGAGATCTGAAGTGGTGGGAGTTAAGGGCAgaagcggca gccaaagaagcagccgctaaggcgCTGCAGTGTATCAAGGTTAACAACTGGGATTTATTCTTCAGCCCG AGTGAAGACAACTTCACCAACGACCTGAACAAAGGTGAAGAAATCACCTCAGATACTAACATCGAAGCA

GCCGAAGAAAACATCTCGCTGGACCTGATCCAGCAGTACTACCTGACCTTTAATTTCGACAACGAGCCG GAAAACATTTCTATCGAAAACCTGAGCTCTGATATCATCGGCCAGCTGGAACTGATGCCGAACATCGAA CGTTTCCCAAACGGTAAAAAGTACGAGCTGGACAAATATACCATGTTCCACTACCTGCGCGCGCAGGAA TTTGAACACGGCAAATCCCGTATCGCACTGACTAACTCCGTTAACGAAGCTCTGCTCAACCCGTCCCGT GTATACACCTTCTTCTCTAGCGACTACGTGAAAAAGGTCAACAAAGCGACTGAAGCTGCAATGTTCTTG

GGTTGGGTTGAACAGCTTGTTTATGATTTTACCGACGAGACGTCCGAAGTATCTACTACCGACAAAATT GCGGATATCACTATCATCATCCCGTACATCGGTCCGGCTCTGAACATTGGCAACATGCTGTACAAAGAC GACTTCGTTGGCGCACTGATCTTCTCCGGTGCGGTGATCCTGCTGGAGTTCATCCCGGAAATCGCCATC CCGGTACTGGGCACCTTTGCTCTGGTTTCTTACATTGCAAACAAGGTTCTGACTGTACAAACCATCGAC AACGCGCTGAGCAAACGTAACGAAAAATGGGATGAAGTTTACAAATATATCGTGACCAACTGGCTGGCT

AAGGTTAATACTCAGATCGACCTCATCCGCAAAAAAATGAAA.GAAGCACTGGAAAACCAGGCGGAAGCT ACCAAGGCAATCATTAACTACCAGTACAACCAGTACACCGAGGAAGAAAAAAACAACATCAACTTCAAC ATCGACGATCTGTCCTCTAAACTGAACGAATCCATCAACAAAGCTATGATCAACATCAACAAGTTCCTG AACCAGTGCTCTGTAAGCTATCTGATGAACTCCATGATCCCGTACGGTGTTAAACGTCTGGAGGACTTC GATGCGTCTCTGAAAGACGCCCTGCTGAAATACATTTACGACAACCGTGGCACTCTGATCGGTCAGGTT

GATCGTCTGAAGGACAAAGTGAACAATACCTTATCGACCGACATCCCTTTTCAGCTCAGTAAATATGTC GATAACCAACGCCTTTTGTCCACTctagaataatgaaagctt

10. Protein sequence of the LHA-CP-EGF fusion

EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT

EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDNNNNNNNNNNDDDDKNSDSECPLSHDGYCLHDGVCMYIEALDKYACNCWGY IGERCQYRDLKWWELRAEAAAKEAAAKALQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSDTNIEAAEE NISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHYLRAQEFEH GKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVSTTDKIADI

TIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNAL SKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDD

- 103 2015268754 16 Dec 2015

LSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL KDKVNNTLSTDIPFQLSKYVDNQRLLST

11. Protein sequence of LH_N/A

EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV

SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ

PDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL

GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL

QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS

TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLST

12. Protein sequence of LH_N/B

PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV

CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN

KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE

NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDEEKLYDDDDKDRWGSSLQCIDVDNEDLFFIADKNSFSDDLSKNER IEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYL

YSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKWEAGLFAGWVKQIVNDFVIEANKSNT MDAIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPWGAFLLESYIDNKNKII KTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSN INIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENKLYL IGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNS

13. Protein sequence of LH_N/C

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG

YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK

TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFWESSGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE

ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKI S DVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVT1LLEAFPEFTIPALGAFVIYSKVQERNE11KTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV

DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFN

14. Protein sequence of LH_N/D

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS

YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI

ERNPALQKLSSESWDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY

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YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI

ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFN

15. Synthesised GnRH peptide pGlu-His-Trp-Ser-Tyr-Gly-Cys-Arg-Pro-Gly-NH2

16. Protein sequence of the LHB-CT-SST28 fusion

PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV

CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN

KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE

NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDGIITSKTKSDDDDKNKALNLQCIDVDNEDLFFIADKNSFSDDLSK NERIEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIF

QYLYSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKWEAGLFAGWVKQIVNDFVIEANK SNTMDKIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPWGAFLLESYIDNKN KIIKTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKE KSNINIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENK LYLIGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALDSAMSN

PAMAPRERKAGCKNFFWKTFTSC

17. Protein sequence of the LHA-CP-SST28 fusion

EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV

SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK

LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKSANSNPAMAPRERKAGCKNFFWKTFTSCALAGGGGSGG GGSGGGGSALVLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSDTNIEAAEENISLDLIQQYYLTFNFD NEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLN PSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNML YKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTN

WLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESINKAMININ KFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRLKDKVNNTLSTDIPFQLS KYVDNQRLLST

18. Protein sequence of the LHD-CT-EGF fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS

YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE

KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN

QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESWDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY

YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVNSDSEC

PLSHDGYCLHDGVCMYIEALDKYACNCWGYIGERCQYRDLKWWELR

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19.

Protein sequence of the LHD-CT-VIP fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK

FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESWDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK

IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVHSDAVF TDNYTRLRKQMAVKKYLNSILN

20. Protein sequence of the LHC-CT-IGF1 fusion

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND

VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFWESSGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ

KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIY SKVQERNE11KTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVGPETLCGAELVD

ALQFVCGDRGFYFNKPTGYGSSSRRAPQTGIVDECCFRSCDLRRLEMYCAPLKPAKSA

21. Protein sequence of the LHD-CT-IGFl fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS

YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI

ERNPALQKLSSESWDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK

ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVGPETLC GAELVDALQFVCGDRGFYFNKPTGYGSSSRRAPQTGIVDECCFRSCDLRRLEMYCAPLKPAKSA

22. Protein sequence of the LHC-CT-VIP fusion

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT

IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANP S SFNKYIGEYKQKLIRKYRFWE S SGEVTVNRNK

FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL

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SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKI S DVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNE11KTID

NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVHSDAVFTDNYTR LRKQMAVKKYLNSILN

23. Protein sequence of the LHC-CT-GnRH fusion

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG

YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK

TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND

VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFWESSGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ

KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVMKPIQKLLAGLI

LLTWCVEGCSSQHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAETQRFECTTHQPRSPLRDLKGALES LIEEETGQKKI

24. Protein sequence of the LHD-CT-GnRH fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK

FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESWDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK

IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSGGGGSALVM KPIQKLLAGLILLTWCVEGCSSQHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAETQRFECTTHQPRS PLRDLKGALESLIEEETGQKKI

25. Protein sequence of the LHD-CT-GRP fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS

YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE

KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESWDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN

KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTKLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI

ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVGNHWAV GHLM

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26. Protein sequence of the LHB-CT-GRP fusion

PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD

KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDEEKLYDDDDKDRWGSSLQCIDVDNEDLFFIADKNSFSDDLSKNER IEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYL YSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKWEAGLFAGWVKQIVNDFVIEANKSNT MDAIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPWGAFLLESYIDNKNKII

KTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSN INIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENKLYL IGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALVGNHWAVGH LM

27. Protein sequence of the LHC-CT-LIF fusion

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND

VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFWESSGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ

KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVSPLPITPVNATC

AIRHPCHNNLMNQIRSQLAQLNGSANALFILYYTAQGEPFPNNLDKLCGPNVTDFPPFHANGTEKAKLV ELYRIWYLGTSLGNITRDQKILNPSALSLHSKLNATADILRGLLSNVLCRLCSKYHVGHVDVTYGPDT SGKDVFQKKKLGCQLLGKYKQIIAVLAQAF

28. Protein sequence of the LHB-CP-LIF fusion

PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV

CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN

KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE

NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP

SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDNNNNNNNNNNDDDDKSPLPITPVNATCAIRHPCHNNLMNQIRSQL AQLNGSANALFILYYTAQGEPFPNNLDKLCGPNVTDFPPFHANGTEKAKLVELYRIWYLGTSLGNITR DQKILNPSALSLHSKLNATADILRGLLSNVLCRLCSKYHVGHVDVTYGPDTSGKDVFQKKKLGCQLLGK

YKQIIAVLAQAFAEAAAKEAAAKALQCIDVDNEDLFFIADKNSFSDDLSKNERIEYNTQSNYIENDFPI NELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYLYSQTFPLDIRDISLTS SFDDALLFSNKVYSFFSMDYIKTANKWEAGLFAGWVKQIVNDFVIEANKSNTMDKIADISLIVPYIGL ALNVGNETAKGNFENAFEIAGASILLEFIPELLIPWGAFLLESYIDNKNKIIKTIDNALTKRNEKWSD MYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSNINIDFNDINSKLNEGI

NQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENKLYLIGSAEYEKSKVNKYLK TIMPFDLSIYTNDTILIEMFNKYNS

29. Protein sequence of the LHC-CT-FGF1 fusion

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK

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TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND

VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT

IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFWESSGEVTVNRNK

FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL

SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ

VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVMFNLPPGNYKKP KLLYCSNGGHFLRILPDGTVDGTRDRSDQHIQLQLSAESVGEVYIKSTETGQYLAMDTDGLLYGSQTPN EECLFLERLEENHYNTYISKKHAEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPLPVSSD

30. Protein sequence of the LHA-CP-FGF1 fusion

EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV

SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ

PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL

GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKMFNLPPGNYKKPKLLYCSNGGHFLRILPDGTVDGTRDR SDQHIQLQLSAESVGEVYIKSTETGQYLAMDTDGLLYGSQTPNEECLFLERLEENHYNTYISKKHAEKN

WFVGLKKNGSCKRGPRTHYGQKAILFLPLPVSSDGGGGSGGGGSGGGGSALVLQCIKVNNWDLFFSPSE DNFTNDLNKGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERF PNGKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGW VEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPV LGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATK

AIINYQYNQYTEEEKNNINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDA SLKDALLKYIYDNRGTLIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLST

31. Protein sequence of the LHA-CT-FGF9 fusion

EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT

EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT

VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTLEGGGGSGGGGSGGGGSGGGGSALVDHLGQSEA GGLPRGPAVTDLDHLKGILRRRQLYCRTGFHLEIFPNGTIQGTRKDHSRFGILEFISIAVGLVSIRGVD SGLYLGMNEKGELYGSEKLTQECVFREQFEENWYNTYSSNLYKHVDTGRRYYVALNKDGTPREGTRTKR

HQKFTHFLPRPVDPDKVPELYKDILSQS

32. Protein sequence of the LHC-CP-FGF9 fusion

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG

YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK

TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT I DL IPKSARKYFEEKALDYYRSIAKRLNSITTANPS SFNKYIGEYKQKLIRKYRFWE S SGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL

SRNPALRKVNPENMLYLFTKFCVDNNNNNNNNNNDDDDKDHLGQSEAGGLPRGPAVTDLDHLKGILRRR QLYCRTGFHLEIFPNGTIQGTRKDHSRFGILEFISIAVGLVSIRGVDSGLYLGMNEKGELYGSEKLTQE

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CVFREQFEENWYNTYSSNLYKHVDTGRRYYVALNKDGTPREGTRTKRHQKFTHFLPRPVDPDKVPELYK

DILSQSAEAAAKEAAAKALQCRELLVKNTDLPFIGDISDVKTDIFLRKDINEETEVIYYPDNVSVDQVI

LSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQKLSDNVEDFTFTRSIEE

ALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKISDVSAIIPYIGPALNIS

NSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTIDNCLEQRIKRWKDSYEWM MGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQVENLKNSLDVKISEAMN NINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEVDKLKAKVNNSFQNTIPF NIFSYTNNSLLKDIINEYFN

33. Protein sequence of the IgA-H_Ntet-CT-SST14 Fusion

ESNQPEKNGTATKPENSGNTTSENGQTEPEKKLELRNVSDIELYSQTNGTYRQHVSLDGIPENTDTYFV

KVKSSAFKDVYIPVASITEEKRNGQSVYKITAKAEKLQQELENKYVDNFTFYLDKKAKEENTNFTSFSN

LVKAINQNPSGTYHLAASLNANEVELGPDERSYIKDTFTGRLIGEKDGKNYAIYNLKKPLFENLSGATV

EKLSLKNVAISGKNDIGSLANEATNGTKIKQVHVDGCVDGIITSKTKSDDDDKNKALNLQCIKIKNEDL TFIAEKNSFSEEPFQDEIVSYNTKNKPLNFNYSLDKIIVDYNLQSKITLPNDRTTPVTKGIPYAPEYKS NAASTIEIHNIDDNTIYQYLYAQKSPTTLQRITMTNSVDDALINSTKIYSYFPSVISKVNQGAQGILFL QWVRDIIDDFTNESSQKTTIDKISDVSTIVPYIGPALNIVKQGYEGNFIGALETTGWLLLEYIPEITL PVIAALSIAESSTQKEKIIKTIDNFLEKRYEKWIEVYKLVKAKWLGTVNTQFQKRSYQMYRSLEYQVDA

IKKIIDYEYKIYSGPDKEQIADEINNLKNKLEEKANKAMININIFMRESSRSFLVNQMINEAKKQLLEF DTQSKNILMQYIKANSKFIGITELKKLESKINKVFSTPIPFSYSKNLDCWVDNEEDIDVLEGGGGSGGG GSGGGGSALVAGCKNFFWKTFTSC

34. Protein sequence of the IgA-H_Ntet-CP-SST14 fusion

ESNQPEKNGTATKPENSGNTTSENGQTEPEKKLELRNVSDIELYSQTNGTYRQHVSLDGIPENTDTYFV

KVKSSAFKDVYIPVASITEEKRNGQSVYKITAKAEKLQQELENKYVDNFTFYLDKKAKEENTNFTSFSN

LVKAINQNPSGTYHLAASLNANEVELGPDERSYIKDTFTGRLIGEKDGKNYAIYNLKKPLFENLSGATV

EKLSLKNVAISGKNDIGSLANEATNGTKIKQVHVDGCVDGIITSKTKSDDDDKAGCKNFFWKTFTSCAL

AGGGGSGGGGSGGGGSALALQCIKIKNEDLTFIAEKNSFSEEPFQDEIVSYNTKNKPLNFNYSLDKIIV DYNLQSKITLPNDRTTPVTKGIPYAPEYKSNAASTIEIHNIDDNTIYQYLYAQKSPTTLQRITMTNSVD DALINSTKIYSYFPSVISKVNQGAQGILFLQWVRDIIDDFTNESSQKTTIDKISDVSTIVPYIGPALNI VKQGYEGNFIGALETTGWLLLEYIPEITLPVIAALSIAESSTQKEKIIKTIDNFLEKRYEKWIEVYKL VKAKWLGTVNTQFQKRSYQMYRSLEYQVDAIKKIIDYEYKIYSGPDKEQIADEINNLKNKLEEKANKAM

ININIFMRESSRSFLVNQMINEAKKQLLEFDTQSKNILMQYIKANSKFIGITELKKLESKINKVFSTPI PFSYSKNLDCWVDNEEDIDV

35. Protein sequence of the LHA-CT-SST14 fusion

EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT

EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT

VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTLEGGGGSGGGGSGGGGSALVAGCKNFFWKTFTS C

36. Protein sequence of the LHA-CT-EGFv3 fusion

EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV

SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ

PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL

GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT

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EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIIT SKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEIT SD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS

TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTLEGGGGSGGGGSGGGGSGGGGSALVDNSDPKCP LSHEGYCLNDGVCMYIGTLDRYACNCWGYVGERCQYRDLKLAELR

37. Protein sequence of the LHE-CT-IL6 fusion

PKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTTPQDFHPPTSLKNGDSSYYD PNYLQSDEEKDRFLKIVTKIFNRINNNLSGGILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNG

SQHILLPNVIIMGAEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDPAL TLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFGGNDLNIITVAQYNDIYTNLLN DYRKIASKLSKVQVSNPQLNPYKDIFQEKYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVK CRETYIGQYKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKIIRFCVDG IITSKTKSLIEGRNKALNLQCIEINNGELFFVASENSYNDDNINTPKEIDDTVTSNNNYENDLDQVILN

FNSESAPGLSDEKLNLTIQNDAYIPKYDSNGTSDIEQHDVNELNVFFYLDAQKVPEGENNVNLTSSIDT ALLEQPKIYTFFSSEFINNVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIWPYIGLALNI GNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKNKVIKAINNALKERDEKWKEV YSFIVSNWMTKINTQFNKRKEQMYQALQNQVNAIKTIIESKYNSYTLEEKNELTNKYDIKQIENELNQK VSIAMNNIDRFLTESSISYLMKIINEVKINKLREYDENVKTYLLNYIIQHGSILGESQQELNSMVTDTL

NNSIPFKLSSYTDDKILISYFNKFFKLEGGGGSGGGGSGGGGSGGGGSALVPPGEDSKDVAAPHRQPLT SSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIIT GLLEFEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWL QDMTTHLILRSFKEFLQSSLRALRQM

38. Protein sequence of the LHB-CT-IL8 fusion

PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV

CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN

KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE

NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDGIITSKTKSLIEGRNKALNLQCIDVDNEDLFFIADKNSFSDDLSK NERIEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIF

QYLYSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKWEAGLFAGWVKQIVNDFVIEANK SNTMDKIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPWGAFLLESYIDNKN KIIKTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKE KSNINIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENK LYLIGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALVAKELR

CQCIKTYSKPFHPKFIKELRVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRWEKFLKRAENS

39. Protein sequence of the LHF-CP-GRAN4 fusion

PVAINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTNPSDFDPPASLKNGSS

AYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGKVLLQEISYAKPYLGNDHTPIDEFSPVTRTTSVNI KLSTNVESSMLLNLLVLGAGPDIFESCCYPVRKLIDPDWYDPSNYGFGSINIVTFSPEYEYTFNDISG GHNSSTESFIADPAISLAHELIHALHGLYGARGVTYEETIEVKQAPLMIAEKPIRLEEFLTFGGQDLNI ITSAMKEKIYNNLLANYEKIATRLSEVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYK KLYSFTESDLANKFKVKARNTYFIKYEFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQSIKLNPKIIDS

IPDKGLVEKIVKFAVENNNNNNNNNNLGCVDGIITSKTKSLIEGRDVKCDMEVSCPDGYTCCRLQSGAW GCCPFTQAVCCEDHIHCCPAGFTCDTQKGTCEALAGGGGSGGGGSGGGGSALVLQCIEVNNSELFFVAS ESSYNENDINTPKEIDDTTNLNNNYENNLDEVILDYNSQTIPQISNIENLNTLVQDNSYVPEYDSNGTS EIEEYDWDFNVFFYLHAQKVPEGETNISLTSSIDTALLEESKDIFFSSEFIDTINKPVNAALFIDWIS KVIRDFTTEATQKSTVDKIADISLIVPYVGLALNIIIEAEKGNFEEAFELLGVGILLEFVPELTIPVIL

VFTIKSYIDSYENKNKAIKAINNSLIEREAKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQNQVDAI

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KTAIEYKYNNYTSDEKNRLESEYNINNIEEELNKKVSLAMKNIERFMTESSISYLMKLINEAKVGKLKK

YDNHVKSDLLNYILDHRSILGEQTNELSDLVTSTLNSSIPFELSSYTNDKILIIYFNRLYKT

40. Protein sequence of the LHD-CP-TGFa fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS

YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE

KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESWDLFTKVCVDGGGGSGGGGSADDDDKWSHFNDCPDSHTQFCFHGTCRFLVQEDK

PACVCHSGYVGARCEHADLLALAGGGGSGGGGSGGGGSALALQCIKVKNNRLPYVADKDSISQEIFENK IITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSYY YLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKKD TLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREKI IKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDKE

NIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNII LVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFN

41. Protein sequence of the LHD-CP-TGFb fusion

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK

FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESWDLFTKVCVDGGGGSGGGGSADDDDKALDTNYCFSSTEKNCCVRQLYIDFRKDLG WKWIHEPKGYHANFCLGPCPYIWSLDTQYSKVLALYNQHNPGASAAPCCVPQALEPLPIVYYVGRKPKV EQLSNMIVRSCKCSALAGGGGSGGGGSGGGGSALALQCIKVKNNRLPYVADKDSISQEIFENKIITDET NVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSYYYLESQK

LSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKKDTLDKIS DVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREKIIKTIEN CLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDKENIKSQV ENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNIILVGEVD RLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFN

42. Protein sequence of the LHB-CT-TNFa fusion

PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV

CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN

KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDEEKLYDDDDKDRWGSSLQCIDVDNEDLFFIADKNSFSDDLSKNER

IEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYL YSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKWEAGLFAGWVKQIVNDFVIEANKSNT MDAIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPWGAFLLESYIDNKNKII KTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSN INIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENKLYL

IGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALVRSSSRTPS DKPVAHWANPQAEGQLQWLNRRANALLANGVELRDNQLWPSEGLYLIYSQVLFKGQGCPSTHVLLTH TISRIAVSYQTKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAES GQVYFGIIAL

43. Protein sequence of the LHD-CT-SDF1 fusion

- 112 2015268754 16 Dec 2015

TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS

YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLWGSPFMGDSSTPEDTFDFTRHTTNIAVE

KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN

QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD

VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFWNIDK FNSLYSDLTNVMSEWYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESWDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNWMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEWEDFTTNIMKK

DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALKPVSLSY RCPCRFFESHVARANVKHLKILNTPNCALQIVARLKNNNRQVCIDPKLKWIQEYLEKALNKRFKM

44. Protein sequence of the LHC-CT-VEGF fusion

PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG

YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK

TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPS SFNKYIGEYKQKLIRKYRFWE S SGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE

ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDWEDFTTNILRKDTLDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV

DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALAPMAEGGGQNHHE VVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMR IKPHQGQHIGEMSFLQHNKCECRPKKDRARQEKKSVRGKGKGQKRKRKKSRYKSWSVYVGARCCLMPWS LPGPHPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLELNERTCRCDKPRR

- 113114

2015268754 19 Dec 2017

Claims (53)

1. CLAIMS:

   1. A method of suppressing or treating cancer by inhibiting autocrine secretion from a cancer cell in a patient, said method comprising administering to the patient an effective amount of a polypeptide comprising:

   (i) a non-cytotoxic protease, which protease is capable of cleaving said SNARE protein expressed in said cancer cell;

   (ii) a Targeting Moiety (TM) that is capable of binding to a Binding Site on a cancer cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the cancer cell; and (iii) a translocation domain that is capable of translocating the protease from within an endosome, across the endosomal membrane and into the cytosol of the cancer cell;

   wherein SNARE protein expression is up-regulated in the cancer cell when compared with SNARE protein expression in the same cell type in a noncancerous state;

   wherein the polypeptide lacks the natural binding function of a clostridial neurotoxin H_cc domain which enables the clostridial neurotoxin to bind to nerve terminals at the neuromuscular junction; and with the proviso that the cancer cell is not a neuroendocrine tumour cell;

   and with the proviso that the polypeptide is not a clostridial neurotoxin (holotoxin).
2. 2. The method according to claim 1, wherein said SNARE protein expression is up-regulated greater than two-fold when compared with SNARE protein expression in the same cell type in a non-cancerous state.
3. 3. The method according to claim 2, wherein said SNARE protein expression is up-regulated greater than two-fold when compared with SNARE protein expression in the same cell type in a non-cancerous state from said patient.
4. 4. A method according to any one of Claims 1 to 3, wherein the cancer is selected from the group consisting of: lung cancer, breast cancer, pancreatic cancer, colorectal cancer, acute leukaemia, bladder cancer, bone cancer, bowel cancer, cervical cancer, chronic lymphocytic leukaemia, Hodgkin's lymphoma,

   115

   2015268754 19 Dec 2017 liver cancer, melanoma skin cancer, oropharyngeal cancer, myeloma, gastric cancer, testicular cancer, or Kaposi sarcoma.
5. 5. A method according to any one of Claims 1 to 4, wherein the cancer cell is selected from the group consisting of:, a lung cancer cell, a breast cancer cell, a pancreatic cancer cell, a colorectal cancer cell, an acute leukaemia cancer cell, a bladder cancer cell, a bone cancer cell, a bowel cancer cell, a cervical cancer cell, a chronic lymphocytic leukaemia cell, a Hodgkin's lymphoma cell, a liver cancer cell, a melanoma skin cancer cell, an oropharyngeal cancer cell, a myeloma cell, a gastric cancer cell, a testicular cancer cell, or a Kaposi sarcoma cancer cell.
6. 6. A method according to any one of the preceding claims, wherein the noncytotoxic protease comprises a clostridial neurotoxin L-chain or an IgA protease.
7. 7. A method according to any one of the preceding claims, wherein the translocation domain comprises a clostridial neurotoxin translocation domain.
8. 8. Use of an effective amount of a polypeptide comprising:

   (i) a non-cytotoxic protease, which protease is capable of cleaving said SNARE protein expressed in said cancer cell;

   (ii) a Targeting Moiety (TM) that is capable of binding to a Binding Site on a cancer cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the cancer cell; and (iii) a translocation domain that is capable of translocating the protease from within an endosome, across the endosomal membrane and into the cytosol of the cancer cell;

   in the preparation of a medicament to suppress or treat cancer by inhibiting autocrine secretion from a cancer cell;

   wherein SNARE protein expression is up-regulated in the cancer cell when compared with SNARE protein expression in the same cell type in a noncancerous state;

   116

   2015268754 19 Dec 2017 wherein the polypeptide lacks the natural binding function of a clostridial neurotoxin H_cc domain which enables the clostridial neurotoxin to bind to nerve terminals at the neuromuscular junction;

   with the proviso that the cancer cell is not a neuroendocrine tumour cell;

   and with the proviso that the polypeptide is not a clostridial neurotoxin (holotoxin).
9. 9. Use according to claim 8, wherein said SNARE protein expression is upregulated greater than two-fold when compared with SNARE protein expression in the same cell type in a non-cancerous state.
10. 10. Use according to claim 9, wherein said SNARE protein expression is upregulated greater than two-fold when compared with SNARE protein expression in the same cell type in a non-cancerous state from said patient.
11. 11. Use according to any one of claims 8 to 10, wherein the cancer is selected from the group consisting of: lung cancer, breast cancer, pancreatic cancer, colorectal cancer, acute leukaemia, bladder cancer, bone cancer, bowel cancer, cervical cancer, chronic lymphocytic leukaemia, Hodgkin's lymphoma, liver cancer, melanoma skin cancer, oropharyngeal cancer, myeloma, gastric cancer, testicular cancer, or Kaposi sarcoma.
12. 12. Use according to any one of Claims 8 to 11, wherein the cancer cell is selected from the group consisting of:, a lung cancer cell, a breast cancer cell, a pancreatic cancer cell, a colorectal cancer cell, an acute leukaemia cancer cell, a bladder cancer cell, a bone cancer cell, a bowel cancer cell, a cervical cancer cell, a chronic lymphocytic leukaemia cell, a Hodgkin's lymphoma cell, a liver cancer cell, a melanoma skin cancer cell, an oropharyngeal cancer cell, a myeloma cell, a gastric cancer cell, a testicular cancer cell, or a Kaposi sarcoma cancer cell.
13. 13. Use according to any one of claims 8 to 12, wherein the non-cytotoxic protease comprises a clostridial neurotoxin L-chain or an IgA protease.

    117
14. 14. Use according to any one of claims 8 to 13, wherein the translocation domain comprises a clostridial neurotoxin translocation domain.

    2015268754 19 Dec 2017

    IPSEN BIOINNOVATION LIMITED

    WATERMARK INTELLECTUAL PROPERTY PTY LTD P42497AU01

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    P31OO3au-pct.txt SEQUENCE LISTING <110> SYNTAXIN LIMITED <120> SUPPRESSION OF CANCER <130> P31003WO <140> PCT/GB2009/051559 <141> 2009-11-17 <150> GB 0820970.2 <151> 2008-11-17 <160> 112 <170> Patentln version 3.5 <210> 1 <211> 2619 <212> DNA <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic polynucleotide

    <400> 1 ggatccatgg agttcgttaa caaacagttc aactataaag acccagttaa cggtgttgac 60 attgcttaca tcaaaatccc gaacgctggc cagatgcagc cggtaaaggc attcaaaatc 120 cacaacaaaa tctgggttat cccggaacgt gataccttta ctaacccgga agaaggtgac 180 ctgaacccgc caccggaagc gaaacaggtg ccggtatctt actatgactc cacctacctg 240 tctaccgata acgaaaagga caactacctg aaaggtgtta ctaaactgtt cgagcgtatt 300 tactccaccg acctgggccg tatgctgctg actagcatcg ttcgcggtat cccgttctgg 360 ggcggttcta ccatcgatac cgaactgaaa gtaatcgaca ctaactgcat caacgttatt 420 cagccggacg gttcctatcg ttccgaagaa ctgaacctgg tgatcatcgg cccgtctgct 480 gatatcatcc agttcgagtg tctgagcttt ggtcacgaag ttctgaacct cacccgtaac 540 ggctacggtt ccactcagta catccgtttc tctccggact tcaccttcgg ttttgaagaa 600 tccctggaag tagacacgaa cccactgctg ggcgctggta aattcgcaac tgatcctgcg 660 gttaccctgg ctcacgaact gattcatgca ggccaccgcc tgtacggtat cgccatcaat 720 ccgaaccgtg tcttcaaagt taacaccaac gcgtattacg agatgtccgg tctggaagtt 780 agcttcgaag aactgcgtac ttttggcggt cacgacgcta aattcatcga ctctctgcaa 840 gaaaacgagt tccgtctgta ctactataac aagttcaaag atatcgcatc caccctgaac 900 aaagcgaaat ccatcgtggg taccactgct tctctccagt acatgaagaa cgtttttaaa 960 gaaaaatacc tgctcagcga agacacctcc ggcaaattct ctgtagacaa gttgaaattc 1020 gataaacttt acaaaatgct gactgaaatt tacaccgaag acaacttcgt taagttcttt 1080 aaagttctga accgcaaaac ctatctgaac ttcgacaagg cagtattcaa aatcaacatc 1140 gtgccgaaag ttaactacac tatctacgat ggtttcaacc Page 1 tgcgtaacac caacctggct 1200

    2015268754 16 Dec 2015

    P31003au-pct.txt

    gctaatttta acggccagaa cacggaaatc aacaacatga acttcacaaa actgaaaaac 1260 ttcactggtc tgttcgagtt ttacaagctg ctgtgcgtcg acggcatcat tacctccaaa 1320 actaaatctg acgatgacga taaaaacaaa gcgctgaacc tgcagtgtat caaggttaac 1380 aactgggatt tattcttcag cccgagtgaa gacaacttca ccaacgacct gaacaaaggt 1440 gaagaaatca cctcagatac taacatcgaa gcagccgaag aaaacatctc gctggacctg 1500 atccagcagt actacctgac ctttaatttc gacaacgagc cggaaaacat ttctatcgaa 1560 aacctgagct ctgatatcat cggccagctg gaactgatgc cgaacatcga acgtttccca 1620 aacggtaaaa agtacgagct ggacaaatat accatgttcc actacctgcg cgcgcaggaa 1680 tttgaacacg gcaaatcccg tatcgcactg actaactccg ttaacgaagc tctgctcaac 1740 ccgtcccgtg tatacacctt cttctctagc gactacgtga aaaaggtcaa caaagcgact 1800 gaagctgcaa tgttcttggg ttgggttgaa cagcttgttt atgattttac cgacgagacg 1860 tccgaagtat ctactaccga caaaattgcg gatatcacta tcatcatccc gtacatcggt 1920 ccggctctga acattggcaa catgctgtac aaagacgact tcgttggcgc actgatcttc 1980 tccggtgcgg tgatcctgct ggagttcatc ccggaaatcg ccatcccggt actgggcacc 2040 tttgctctgg tttcttacat tgcaaacaag gttctgactg tacaaaccat cgacaacgcg 2100 ctgagcaaac gtaacgaaaa atgggatgaa gtttacaaat atatcgtgac caactggctg 2160 gctaaggtta atactcagat cgacctcatc cgcaaaaaaa tgaaagaagc actggaaaac 2220 caggcggaag ctaccaaggc aatcattaac taccagtaca accagtacac cgaggaagaa 2280 aaaaacaaca tcaacttcaa catcgacgat ctgtcctcta aactgaacga atccatcaac 2340 aaagctatga tcaacatcaa caagttcctg aaccagtgct ctgtaagcta tctgatgaac 2400 tccatgatcc cgtacggtgt taaacgtctg gaggacttcg atgcgtctct gaaagacgcc 2460 ctgctgaaat acatttacga caaccgtggc actctgatcg gtcaggttga tcgtctgaag 2520 gacaaagtga acaatacctt atcgaccgac atcccttttc agctcagtaa atatgtcgat 2580 aaccaacgcc ttttgtccac tctagaataa tgaaagctt 2619 <210> 2 <211> 2643 <212> DNA <213> Artificial Sequence <220> <221> source <223> /note=Description of Artificial Sequence: synthetic polynucleotide <400> 2 ggatccatgc cggttaccat caacaacttc aactacaacg acccgatcga caacaacaac 60 atcattatga tggaaccgcc gttcgcacgt ggtaccggac gttactacaa ggcttttaag 120 atcaccgacc gtatctggat catcccggaa cgttacacct tcggttacaa acctgaggac 180 ttcaacaaga gtagcgggat tttcaatcgt gacgtctgcg agtactatga tccagattat 240

    Page 2

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    ctgaatacca acgataagaa gaacatattc cttcagacta tgattaaact cttcaaccgt 300 atcaaaagca aaccgctcgg tgaaaaactc ctcgaaatga ttatcaacgg tatcccgtac 360 ctcggtgacc gtcgtgtccc gcttgaagag ttcaacacca acatcgcaag cgtcaccgtc 420 aacaaactca tcagcaaccc aggtgaagtc gaacgtaaaa aaggtatctt cgcaaacctc 480 atcatcttcg gtccgggtcc ggtcctcaac gaaaacgaaa ccatcgacat cggtatccag 540 aaccacttcg caagccgtga aggtttcggt ggtatcatgc agatgaaatt ctgcccggaa 600 tacgtcagtg tcttcaacaa cgtccaggaa aacaaaggtg caagcatctt caaccgtcgt 660 ggttacttca gcgacccggc actcatcctc atgcatgaac tcatccacgt cctccacggt 720 ctctacggta tcaaagttga cgacctcccg atcgtcccga acgagaagaa attcttcatg 780 cagagcaccg acgcaatcca ggctgaggaa ctctacacct tcggtggcca agacccaagt 840 atcataaccc cgtccaccga caaaagcatc tacgacaaag tcctccagaa cttcaggggt 900 atcgtggaca gactcaacaa agtcctcgtc tgcatcagcg acccgaacat caatatcaac 960 atatacaaga acaagttcaa agacaagtac aaattcgtcg aggacagcga aggcaaatac 1020 agcatcgacg tagaaagttt cgacaagctc tacaaaagcc tcatgttcgg tttcaccgaa 1080 accaacatcg ccgagaacta caagatcaag acaagggcaa gttacttcag cgacagcctc 1140 ccgcctgtca aaatcaagaa cctcttagac aacgagattt acacaattga agagggcttc 1200 aacatcagtg acaaagacat ggagaaggaa tacagaggtc agaacaaggc tatcaacaaa 1260 caggcatacg aggagatcag caaagaacac ctcgcagtct acaagatcca gatgtgcgtc 1320 gacggcatca ttacctccaa aactaaatct gacgatgacg ataaaaacaa agcgctgaac 1380 ctgcagtgca tcgacgttga caacgaagac ctgttcttca tcgctgacaa aaacagcttc 1440 agtgacgacc tgagcaaaaa cgaacgtatc gaatacaaca cccagagcaa ctacatcgaa 1500 aacgacttcc cgatcaacga actgatcctg gacaccgacc tgataagtaa aatcgaactg 1560 ccgagcgaaa acaccgaaag tctgaccgac ttcaacgttg acgttccggt ttacgaaaaa 1620 cagccggcta tcaagaaaat cttcaccgac gaaaacacca tcttccagta cctgtacagc 1680 cagaccttcc cgctggacat ccgtgacatc agtctgacca gcagtttcga cgacgctctg 1740 ctgttcagca acaaagttta cagtttcttc agcatggact acatcaaaac cgctaacaaa 1800 gttgttgaag cagggctgtt cgctggttgg gttaaacaga tcgttaacga cttcgttatc 1860 gaagctaaca aaagcaacac tatggacaaa atcgctgaca tcagtctgat cgttccgtac 1920 atcggtctgg ctctgaacgt tggtaacgaa accgctaaag gtaactttga aaacgctttc 1980 gagatcgctg gtgcaagcat cctgctggag ttcatcccgg aactgctgat cccggttgtt 2040 ggtgctttcc tgctggaaag ttacatcgac aacaaaaaca agatcatcaa aaccatcgac 2100 aacgctctga ccaaacgtaa cgaaaaatgg agtgatatgt acggtctgat cgttgctcag 2160 tggctgagca ccgtcaacac ccagttctac accatcaaag aaggtatgta caaagctctg 2220 aactaccagg ctcaggctct ggaagagatc atcaaatacc Page 3 gttacaacat ctacagtgag 2280

    P31OO3au-pct. txt

    2015268754 16 Dec 2015 aaggaaaaga gtaacatcaa catcgacttc aacgacatca acagcaaact gaacgaaggt 2340 atcaaccagg ctatcgacaa catcaacaac ttcatcaacg gttgcagtgt tagctacctg 2400 atgaagaaga tgatcccgct ggctgttgaa aaactgctgg acttcgacaa caccctgaaa 2460 aagaacctgc tgaactacat cgacgaaaac aagctgtacc tgatcggtag tgctgaatac 2520 gaaaaaagta aagtgaacaa atacctgaag accatcatgc cgttcgacct gagtatctac 2580 accaacgaca ccatcctgat cgaaatgttc aacaaataca actctctaga ataatgaaag 2640 ctt 2643 <210> 3 <211> 2622 <212> DNA <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: synthetic polynucleotide

    <400> 3 ggatccatgc cgatcaccat caacaacttc aactacagcg atccggtgga taacaaaaac 60 atcctgtacc tggataccca tctgaatacc ctggcgaacg aaccggaaaa agcgtttcgt 120 atcaccggca acatttgggt tattccggat cgttttagcc gtaacagcaa cccgaatctg 180 aataaaccgc cgcgtgttac cagcccgaaa agcggttatt acgatccgaa ctatctgagc 240 accgatagcg ataaagatac cttcctgaaa gaaatcatca aactgttcaa acgcatcaac 300 agccgtgaaa ttggcgaaga actgatctat cgcctgagca ccgatattcc gtttccgggc 360 aacaacaaca ccccgatcaa cacctttgat ttcgatgtgg atttcaacag cgttgatgtt 420 aaaacccgcc agggtaacaa ttgggtgaaa accggcagca ttaacccgag cgtgattatt 480 accggtccgc gcgaaaacat tattgatccg gaaaccagca cctttaaact gaccaacaac 540 acctttgcgg cgcaggaagg ttttggcgcg ctgagcatta ttagcattag cccgcgcttt 600 atgctgacct atagcaacgc gaccaacgat gttggtgaag gccgtttcag caaaagcgaa 660 ttttgcatgg acccgatcct gatcctgatg catgaactga accatgcgat gcataacctg 720 tatggcatcg cgattccgaa cgatcagacc attagcagcg tgaccagcaa catcttttac 780 agccagtaca acgtgaaact ggaatatgcg gaaatctatg cgtttggcgg tccgaccatt 840 gatctgattc cgaaaagcgc gcgcaaatac ttcgaagaaa aagcgctgga ttactatcgc 900 agcattgcga aacgtctgaa cagcattacc accgcgaatc cgagcagctt caacaaatat 960 atcggcgaat ataaacagaa actgatccgc aaatatcgct ttgtggtgga aagcagcggc 1020 gaagttaccg ttaaccgcaa taaattcgtg gaactgtaca acgaactgac ccagatcttc 1080 accgaattta actatgcgaa aatctataac gtgcagaacc gtaaaatcta cctgagcaac 1140 gtgtataccc cggtgaccgc gaatattctg gatgataacg tgtacgatat ccagaacggc 1200 tttaacatcc cgaaaagcaa cctgaacgtt ctgtttatgg Page 4 gccagaacct gagccgtaat 1260

    2015268754 16 Dec 2015

    P31003au-pct.txt

    ccggcgctgc gtaaagtgaa cccggaaaac atgctgtacc tgttcaccaa attttgcgtc 1320 gacgcgattg atggtcgtag cctgtacaac aaaaccctgc agtgtcgtga actgctggtg 1380 aaaaacaccg atctgccgtt tattggcgat atcagcgatg tgaaaaccga tatcttcctg 1440 cgcaaagata tcaacgaaga aaccgaagtg atctactacc cggataacgt gagcgttgat 1500 caggtgatcc tgagcaaaaa caccagcgaa catggtcagc tggatctgct gtatccgagc 1560 attgatagcg aaagcgaaat tctgccgggc gaaaaccagg tgttttacga taaccgtacc 1620 cagaacgtgg attacctgaa cagctattac tacctggaaa gccagaaact gagcgataac 1680 gtggaagatt ttacctttac ccgcagcatt gaagaagcgc tggataacag cgcgaaagtt 1740 tacacctatt ttccgaccct ggcgaacaaa gttaatgcgg gtgttcaggg cggtctgttt 1800 ctgatgtggg cgaacgatgt ggtggaagat ttcaccacca acatcctgcg taaagatacc 1860 ctggataaaa tcagcgatgt tagcgcgatt attccgtata ttggtccggc gctgaacatt 1920 agcaatagcg tgcgtcgtgg caattttacc gaagcgtttg cggttaccgg tgtgaccatt 1980 ctgctggaag cgtttccgga atttaccatt ccggcgctgg gtgcgtttgt gatctatagc 2040 aaagtgcagg aacgcaacga aatcatcaaa accatcgata actgcctgga acagcgtatt 2100 aaacgctgga aagatagcta tgaatggatg atgggcacct ggctgagccg tattatcacc 2160 cagttcaaca acatcagcta ccagatgtac gatagcctga actatcaggc gggtgcgatt 2220 aaagcgaaaa tcgatctgga atacaaaaaa tacagcggca gcgataaaga aaacatcaaa 2280 agccaggttg aaaacctgaa aaacagcctg gatgtgaaaa ttagcgaagc gatgaataac 2340 atcaacaaat tcatccgcga atgcagcgtg acctacctgt tcaaaaacat gctgccgaaa 2400 gtgatcgatg aactgaacga atttgatcgc aacaccaaag cgaaactgat caacctgatc 2460 gatagccaca acattattct ggtgggcgaa gtggataaac tgaaagcgaa agttaacaac 2520 agcttccaga acaccatccc gtttaacatc ttcagctata ccaacaacag cctgctgaaa 2580 gatatcatca acgaatactt caatctagaa taatgaaagc tt 2622

    <210> 4 <211> 2638 <212> DNA <213> Artificial Sequence <220>

    <221> source <223> /note=”Description of Artificial Sequence: Synthetic polynucleotide <400> 4 ggatccatga cgtggccagt taaggatttc aactactcag atcctgtaaa tgacaacgat 60 attctgtacc ttcgcattcc acaaaataaa ctgatcacca caccagtcaa agcattcatg 120 attactcaaa acatttgggt cattccagaa cgcttttcta gtgacacaaa tccgagttta 180 tctaaacctc cgcgtccgac gtccaaatat cagagctatt acgatccctc atatctcagt 240 acggacgaac aaaaagatac tttccttaaa ggtatcatta aactgtttaa gcgtattaat 300

    Page 5

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    gagcgcgata tcgggaaaaa gttgattaat tatcttgttg tgggttcccc gttcatgggc 360 gatagctcta cccccgaaga cacttttgat tttacccgtc atacgacaaa catcgcggta 420 gagaagtttg agaacggatc gtggaaagtc acaaacatca ttacacctag cgtcttaatt 480 tttggtccgc tgccaaacat cttagattat acagccagcc tgactttgca ggggcaacag 540 tcgaatccga gtttcgaagg ttttggtacc ctgagcattc tgaaagttgc cccggaattt 600 ctgctcactt tttcagatgt caccagcaac cagagctcag cagtattagg aaagtcaatt 660 ttttgcatgg acccggttat tgcactgatg cacgaactga cgcactctct gcatcaactg 720 tatgggatca acatccccag tgacaaacgt attcgtcccc aggtgtctga aggatttttc 780 tcacaggatg ggccgaacgt ccagttcgaa gagttgtata ctttcggagg cctggacgta 840 gagatcattc cccagattga gcgcagtcag ctgcgtgaga aggcattggg ccattataag 900 gatattgcaa aacgcctgaa taacattaac aaaacgattc catcttcgtg gatctcgaat 960 attgataaat ataagaaaat ttttagcgag aaatataatt ttgataaaga taatacaggt 1020 aactttgtgg ttaacattga caaattcaac tccctttaca gtgatttgac gaatgtaatg 1080 agcgaagttg tgtatagttc ccaatacaac gttaagaatc gtacccatta cttctctcgt 1140 cactacctgc cggttttcgc gaacatcctt gacgataata tttacactat tcgtgacggc 1200 tttaacttga ccaacaaggg cttcaatatt gaaaattcag gccagaacat tgaacgcaac 1260 ccggccttgc agaaactgtc gagtgaatcc gtggttgacc tgtttaccaa agtctgcgtc 1320 gacaaaagcg aagagaagct gtacgatgac gatgacaaag atcgttgggg atcgtccctg 1380 cagtgtatta aagtgaaaaa caatcggctg ccttatgtag cagataaaga tagcattagt 1440 caggagattt tcgaaaataa aattatcact gacgaaacca atgttcagaa ttattcagat 1500 aaattttcac tggacgaaag catcttagat ggccaagttc cgattaaccc ggaaattgtt 1560 gatccgttac tgccgaacgt gaatatggaa ccgttaaacc tccctggcga agagatcgta 1620 ttttatgatg acattacgaa atatgtggac taccttaatt cttattacta tttggaaagc 1680 cagaaactgt ccaataacgt ggaaaacatt actctgacca caagcgtgga agaggcttta 1740 ggctactcaa ataagattta taccttcctc ccgtcgctgg cggaaaaagt aaataaaggt 1800 gtgcaggctg gtctgttcct caactgggcg aatgaagttg tcgaagactt taccacgaat 1860 attatgaaaa aggataccct ggataaaatc tccgacgtct cggttattat cccatatatt 1920 ggccctgcgt taaatatcgg taatagtgcg ctgcggggga attttaacca ggcctttgct 1980 accgcgggcg tcgcgttcct cctggagggc tttcctgaat ttactatccc ggcgctcggt 2040 gtttttacat tttactcttc catccaggag cgtgagaaaa ttatcaaaac catcgaaaac 2100 tgcctggagc agcgggtgaa acgctggaaa gattcttatc aatggatggt gtcaaactgg 2160 ttatctcgca tcacgaccca attcaaccat attaattacc agatgtatga tagtctgtcg 2220 taccaagctg acgccattaa agccaaaatt gatctggaat ataaaaagta ctctggtagc 2280 gataaggaga acatcaaaag ccaggtggag aaccttaaga Page 6 atagtctgga tgtgaaaatc 2340

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 tctgaagcta tgaataacat taacaaattc attcgtgaat gttcggtgac gtacctgttc 2400 aagaatatgc tgccaaaagt tattgatgaa ctgaataaat ttgatctgcg taccaaaacc 2460 gaacttatca acctcatcga ctcccacaac attatccttg tgggcgaagt ggatcgtctg 2520 aaggccaaag taaacgagag ctttgaaaat acgatgccgt ttaatatttt ttcatatacc 2580 aataactcct tgctgaaaga tatcatcaat gaatatttca atctagatta ataagctt 2638 <210> 5 <211> 195 <212> DNA <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: synthetic polynucleotide <400> 5 ggatccgtcg acctgcaggg tctagaaggc ggtggcggta gcggcggtgg cggtagcggc 60 ggtggcggta gcggcggtgg cggtagcgca ctagtgcagg aaagacctcc attacaacaa 120 cctccacatc gcgataagaa accatgtaag aatttctttt ggaaaacatt tagcagttgc 180 aaatgataaa agctt 195 <210> 6 <211> 2779 <212> DNA <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic polynucleotide <400> 6 ggatccatga cgtggccagt taaggatttc aactactcag atcctgtaaa tgacaacgat 60 attctgtacc ttcgcattcc acaaaataaa ctgatcacca caccagtcaa agcattcatg 120 attactcaaa acatttgggt cattccagaa cgcttttcta gtgacacaaa tccgagttta 180 tctaaacctc cgcgtccgac gtccaaatat cagagctatt acgatccctc atatctcagt 240 acggacgaac aaaaagatac tttccttaaa ggtatcatta aactgtttaa gcgtattaat 300 gagcgcgata tcgggaaaaa gttgattaat tatcttgttg tgggttcccc gttcatgggc 360 gatagctcta cccccgaaga cacttttgat tttacccgtc atacgacaaa catcgcggta 420 gagaagtttg agaacggatc gtggaaagtc acaaacatca ttacacctag cgtcttaatt 480 tttggtccgc tgccaaacat cttagattat acagccagcc tgactttgca ggggcaacag 540 tcgaatccga gtttcgaagg ttttggtacc ctgagcattc tgaaagttgc cccggaattt 600 ctgctcactt tttcagatgt caccagcaac cagagctcag cagtattagg aaagtcaatt 660 ttttgcatgg acccggttat tgcactgatg cacgaactga cgcactctct gcatcaactg 720 tatgggatca acatccccag tgacaaacgt attcgtcccc aggtgtctga aggatttttc 780

    Page 7

    2015268754 16 Dec 2015

    P31003au-pct.txt

    tcacaggatg ggccgaacgt ccagttcgaa gagttgtata ctttcggagg cctggacgta 840 gagatcattc cccagattga gcgcagtcag ctgcgtgaga aggcattggg ccattataag 900 gatattgcaa aacgcctgaa taacattaac aaaacgattc catcttcgtg gatctcgaat 960 attgataaat ataagaaaat ttttagcgag aaatataatt ttgataaaga taatacaggt 1020 aactttgtgg ttaacattga caaattcaac tccctttaca gtgatttgac gaatgtaatg 1080 agcgaagttg tgtatagttc ccaatacaac gttaagaatc gtacccatta cttctctcgt 1140 cactacctgc cggttttcgc gaacatcctt gacgataata tttacactat tcgtgacggc 1200 tttaacttga ccaacaaggg cttcaatatt gaaaattcag gccagaacat tgaacgcaac 1260 ccggccttgc agaaactgtc gagtgaatcc gtggttgacc tgtttaccaa agtctgcgtc 1320 gacaaaagcg aagagaagct gtacgatgac gatgacaaag atcgttgggg atcgtccctg 1380 cagtgtatta aagtgaaaaa caatcggctg ccttatgtag cagataaaga tagcattagt 1440 caggagattt tcgaaaataa aattatcact gacgaaacca atgttcagaa ttattcagat 1500 aaattttcac tggacgaaag catcttagat ggccaagttc cgattaaccc ggaaattgtt 1560 gatccgttac tgccgaacgt gaatatggaa ccgttaaacc tccctggcga agagatcgta 1620 ttttatgatg acattacgaa atatgtggac taccttaatt cttattacta tttggaaagc 1680 cagaaactgt ccaataacgt ggaaaacatt actctgacca caagcgtgga agaggcttta 1740 ggctactcaa ataagattta taccttcctc ccgtcgctgg cggaaaaagt aaataaaggt 1800 gtgcaggctg gtctgttcct caactgggcg aatgaagttg tcgaagactt taccacgaat 1860 attatgaaaa aggataccct ggataaaatc tccgacgtct cggttattat cccatatatt 1920 ggccctgcgt taaatatcgg taatagtgcg ctgcggggga attttaacca ggcctttgct 1980 accgcgggcg tcgcgttcct cctggagggc tttcctgaat ttactatccc ggcgctcggt 2040 gtttttacat tttactcttc catccaggag cgtgagaaaa ttatcaaaac catcgaaaac 2100 tgcctggagc agcgggtgaa acgctggaaa gattcttatc aatggatggt gtcaaactgg 2160 ttatctcgca tcacgaccca attcaaccat attaattacc agatgtatga tagtctgtcg 2220 taccaagctg acgccattaa agccaaaatt gatctggaat ataaaaagta ctctggtagc 2280 gataaggaga acatcaaaag ccaggtggag aaccttaaga atagtctgga tgtgaaaatc 2340 tctgaagcta tgaataacat taacaaattc attcgtgaat gttcggtgac gtacctgttc 2400 aagaatatgc tgccaaaagt tattgatgaa ctgaataaat ttgatctgcg taccaaaacc 2460 gaacttatca acctcatcga ctcccacaac attatccttg tgggcgaagt ggatcgtctg 2520 aaggccaaag taaacgagag ctttgaaaat acgatgccgt ttaatatttt ttcatatacc 2580 aataactcct tgctgaaaga tatcatcaat gaatatttca atctagaagg cggtggcggt 2640 agcggcggtg gcggtagcgg cggtggcggt agcgcactag tgcaggaaag acctccatta 2700 caacaacctc cacatcgcga taagaaacca tgtaagaatt tcttttggaa aacatttagc 2760

    agttgcaaat aataagctt

    2779

    Page 8

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <210> 7 <211> 920 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: synthetic polypeptide <400> 7

    Thr Trp Pro 1 Val Lys 5 Asp Phe Asn Tyr Ser Asp Pro val 10 Asn Asp 15 Asn Asp lie Leu Tyr Leu Arg Ile Pro Gin Asn Lys Leu Ile Thr Thr Pro 20 25 30 Val Lys Ala Phe Met Ile Thr Gin Asn lie Trp val Ile Pro Glu Arg 35 40 45 Phe Ser Ser Asp Thr Asn Pro ser Leu Ser Lys pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gin Ser Tyr Tyr Asp Pro ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 Gin Lys Asp Thr phe Leu Lys Gly Ile lie Lys Leu Phe Lys Arg lie 85 90 95 Asn Glu Arg Asp lie Gly Lys Lys Leu lie Asn Tyr Leu Val val Gly 100 105 110 Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg His Thr Thr Asn lie Ala Val Glu Lys Phe Glu Asn Gly Ser 130 135 140 Trp Lys Val Thr Asn lie lie Thr Pro Ser val Leu lie Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu ASp Tyr Thr Ala ser Leu Thr Leu Gin Gly Gin 165 170 175 Gin ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu Lys 180 185 190 Val Ala Pro Glu Phe Leu Leu Thr Phe ser Asp val Thr Ser Asn Gin 195 200 205 Ser ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met ASp Pro Val Ile 210 215 220

    Page 9

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Ala 225 Leu Met His Glu Leu 230 Thr Hi s Ser Leu His 235 Gin Leu Tyr Gly lie 240 Asn lie pro ser Asp Lys Arg He Arg Pro Gin val Ser Glu Gly Phe 245 250 255 Phe Ser Gin Asp Gly Pro Asn Val Gin Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu ASp val Glu lie He Pro Gin lie Glu Arg Ser Gin Leu 275 280 285 Arg Glu Lys Ala Leu Gly His Tyr Lys Asp lie Ala Lys Arg Leu Asn 290 295 300 Asn lie Asn Lys Thr lie Pro Ser ser Trp lie Ser Asn lie Asp Lys 305 310 315 320 Tyr Lys Lys lie Phe ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Gly Asn Phe val val Asn lie Asp Lys Phe Asn Ser Leu Tyr Ser Asp 340 345 350 Leu Thr Asn val Met ser Glu Val Val Tyr ser Ser Gin Tyr Asn Val 355 360 365 Lys Asn Arg Thr Hi s Tyr Phe Ser Arg Hi s Tyr Leu Pro val Phe Ala 370 375 380 Asn lie Leu Asp Asp Asn He Tyr Thr lie Arg ASp Gly Phe Asn Leu 385 390 395 400 Thr Asn Lys Gly Phe Asn He Gl u Asn Ser Gly Gin Asn He Glu Arg 405 410 415 Asn Pro Ala Leu Gin Lys Leu Ser Ser Glu Ser val val Asp Leu Phe 420 425 430 Thr Lys val cys Val Asp Lys Ser Glu Glu Lys Leu Tyr Asp Asp Asp 435 440 445 Asp Lys Asp Arg Trp Gly ser Ser Leu Gin Cys lie Lys Val Lys Asn 450 455 460 Asn Arg Leu Pro Tyr val Ala Asp Lys Asp Ser lie ser Gin Glu lie 465 470 475 480 Phe Glu Asn Lys lie lie Thr Asp Glu Thr Asn Val Gin Asn Tyr Ser 485 490 495

    Page 10

    P31OO3au-pct.txt

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    Asp Lys Phe Ser 500 Leu Asp Glu Ser lie 505 Leu Asp Gly Gln val 510 Pro Ile Asn Pro Glu ile val Asp Pro Leu Leu pro Asn val Asn Met Glu Pro 515 520 525 Leu Asn Leu pro Gly Glu Glu Ile Val Phe Tyr Asp Asp Ile Thr Lys 530 535 540 Tyr val Asp Tyr Leu Asn Ser Tyr Tyr Tyr Leu Glu Ser Gln Lys Leu 545 550 555 560 ser Asn Asn val Glu Asn lie Thr Leu Thr Thr Ser val Glu Glu Ala 565 570 575 Leu Gly Tyr Ser Asn Lys Ile Tyr Thr Phe Leu Pro Ser Leu Ala Glu 580 585 590 Lys val Asn Lys Gly val Gln Ala Gly Leu Phe Leu Asn Trp Ala Asn 595 600 605 Glu Val Val Glu Asp Phe Thr Thr Asn lie Met Lys Lys Asp Thr Leu 610 615 620 Asp Lys lie Ser Asp val Ser val lie lie Pro Tyr lie Gly Pro Ala 625 630 635 640 Leu Asn Ile Gly Asn ser Ala Leu Arg Gly Asn Phe Asn Gln Ala Phe 645 650 655 Ala Thr Ala Gly val Ala Phe Leu Leu Glu Gly Phe Pro Glu Phe Thr 660 665 670 Ile Pro Ala Leu Gly Val Phe Thr Phe Tyr Ser Ser Ile Gln Glu Arg 675 680 685 Glu Lys Ile ile Lys Thr lie Glu Asn Cys Leu Glu Gln Arg val Lys 690 695 700 Arg Trp Lys Asp ser Tyr Gln Trp Met val Ser Asn Trp Leu Ser Arg 705 710 715 720 He Thr Thr Gln Phe Asn His lie Asn Tyr Gln Met Tyr Asp ser Leu 725 730 735 ser Tyr Gln Ala Asp Ala lie Lys Ala Lys lie Asp Leu Glu Tyr Lys 740 745 750 Lys Tyr Ser Gly ser Asp Lys Glu Asn Ile Lys Ser Gln val Glu Asn 755 760 765

    Page 11

    P31OO3au-pct.txt

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    Leu Lys Asn Ser Leu Asp Val 775 Lys ile Ser Glu Ala Met 780 Asn Asn lie 770 Asn Lys Phe Ile Arg Glu cys ser Val Thr Tyr Leu Phe Lys Asn Met 785 790 795 800 Leu Pro Lys val Ile Asp Glu Leu Asn Lys Phe Asp Leu Arg Thr Lys 805 810 815 Thr Glu Leu lie Asn Leu Ile Asp Ser Hi s Asn lie lie Leu val Gly 820 825 830 Glu val Asp Arg Leu Lys Ala Lys val Asn Glu Ser Phe Glu Asn Thr 835 840 845 Met Pro Phe Asn Tie Phe Ser Tyr Thr Asn Asn ser Leu Leu Lys Asp 850 855 860 Ile Ile Asn Glu Tyr Phe Asn Leu Glu Gly Gly Gly Gly ser Gly Gly 865 870 875 880 Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Val Gin Glu Arg pro Pro 885 890 895 Leu Gin Gin Pro pro Hi s Arg Asp Lys Lys pro Cys Lys Asn Phe Phe 900 905 910 Trp Lys Thr Phe Ser Ser Cys Lys 915 920

    <210> 8 <211> 277 <212> DNA <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polynucleotide <400> 8 ggatccgtcg acaacaacaa taacaacaac aataacaaca acgacgatga cgataaaaat 60 tcagatagcg aatgtccact tagtcacgac gggtactgtt tgcatgatgg tgtgtgtatg 120 tatatagaag cactagacaa atacgcttgc aattgcgtag ttggctatat aggagagcga 180 tgccaatata gagatctgaa gtggtgggag ttaagggcag aagcggcagc caaagaagca 240 gccgctaagg cgctgcagag tctagaataa taagctt 277 <210> 9 <211> 2802 <212> DNA <213> Artificial Sequence page 12

    2015268754 16 Dec 2015

    P31003au-pct.txt <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polynucleotide

    <400> 9 ggatccatgg agttcgttaa caaacagttc aactataaag acccagttaa cggtgttgac 60 attgcttaca tcaaaatccc gaacgctggc cagatgcagc cggtaaaggc attcaaaatc 120 cacaacaaaa tctgggttat cccggaacgt gataccttta ctaacccgga agaaggtgac 180 ctgaacccgc caccggaagc gaaacaggtg ccggtatctt actatgactc cacctacctg 240 tctaccgata acgaaaagga caactacctg aaaggtgtta ctaaactgtt cgagcgtatt 300 tactccaccg acctgggccg tatgctgctg actagcatcg ttcgcggtat cccgttctgg 360 ggcggttcta ccatcgatac cgaactgaaa gtaatcgaca ctaactgcat caacgttatt 420 cagccggacg gttcctatcg ttccgaagaa ctgaacctgg tgatcatcgg cccgtctgct 480 gatatcatcc agttcgagtg tctgagcttt ggtcacgaag ttctgaacct cacccgtaac 540 ggctacggtt ccactcagta catccgtttc tctccggact tcaccttcgg ttttgaagaa 600 tccctggaag tagacacgaa cccactgctg ggcgctggta aattcgcaac tgatcctgcg 660 gttaccctgg ctcacgaact gattcatgca ggccaccgcc tgtacggtat cgccatcaat 720 ccgaaccgtg tcttcaaagt taacaccaac gcgtattacg agatgtccgg tctggaagtt 780 agcttcgaag aactgcgtac ttttggcggt cacgacgcta aattcatcga ctctctgcaa 840 gaaaacgagt tccgtctgta ctactataac aagttcaaag atatcgcatc caccctgaac 900 aaagcgaaat ccatcgtggg taccactgct tctctccagt acatgaagaa cgtttttaaa 960 gaaaaatacc tgctcagcga agacacctcc ggcaaattct ctgtagacaa gttgaaattc 1020 gataaacttt acaaaatgct gactgaaatt tacaccgaag acaacttcgt taagttcttt 1080 aaagttctga accgcaaaac ctatctgaac ttcgacaagg cagtattcaa aatcaacatc 1140 gtgccgaaag ttaactacac tatctacgat ggtttcaacc tgcgtaacac caacctggct 1200 gctaatttta acggccagaa cacggaaatc aacaacatga acttcacaaa actgaaaaac 1260 ttcactggtc tgttcgagtt ttacaagctg ctgtgcgtcg acaacaacaa taacaacaac 1320 aataacaaca acgacgatga cgataaaaat tcagatagcg aatgtccact tagtcacgac 1380 gggtactgtt tgcatgatgg tgtgtgtatg tatatagaag cactagacaa atacgcttgc 1440 aattgcgtag ttggctatat aggagagcga tgccaatata gagatctgaa gtggtgggag 1500 ttaagggcag aagcggcagc caaagaagca gccgctaagg cgctgcagtg tatcaaggtt 1560 aacaactggg atttattctt cagcccgagt gaagacaact tcaccaacga cctgaacaaa 1620 ggtgaagaaa tcacctcaga tactaacatc gaagcagccg aagaaaacat ctcgctggac 1680 ctgatccagc agtactacct gacctttaat ttcgacaacg agccggaaaa catttctatc 1740 gaaaacctga gctctgatat catcggccag ctggaactga tgccgaacat cgaacgtttc 1800 ccaaacggta aaaagtacga gctggacaaa tataccatgt tccactacct gcgcgcgcag 1860

    Page 13 p31OO3au-pct.txt

    2015268754 16 Dec 2015

    gaatttgaac acggcaaatc ccgtatcgca ctgactaact ccgttaacga agctctgctc 1920 aacccgtccc gtgtatacac cttcttctct agcgactacg tgaaaaaggt caacaaagcg 1980 actgaagctg caatgttctt gggttgggtt gaacagcttg tttatgattt taccgacgag 2040 acgtccgaag tatctactac cgacaaaatt gcggatatca ctatcatcat cccgtacatc 2100 ggtccggctc tgaacattgg caacatgctg tacaaagacg acttcgttgg cgcactgatc 2160 ttctccggtg cggtgatcct gctggagttc atcccggaaa tcgccatccc ggtactgggc 2220 acctttgctc tggtttctta cattgcaaac aaggttctga ctgtacaaac catcgacaac 2280 gcgctgagca aacgtaacga aaaatgggat gaagtttaca aatatatcgt gaccaactgg 2340 ctggctaagg ttaatactca gatcgacctc atccgcaaaa aaatgaaaga agcactggaa 2400 aaccaggcgg aagctaccaa ggcaatcatt aactaccagt acaaccagta caccgaggaa 2460 gaaaaaaaca acatcaactt caacatcgac gatctgtcct ctaaactgaa cgaatccatc 2520 aacaaagcta tgatcaacat caacaagttc ctgaaccagt gctctgtaag ctatctgatg 2580 aactccatga tcccgtacgg tgttaaacgt ctggaggact tcgatgcgtc tctgaaagac 2640 gccctgctga aatacattta cgacaaccgt ggcactctga tcggtcaggt tgatcgtctg 2700 aaggacaaag tgaacaatac cttatcgacc gacatccctt ttcagctcag taaatatgtc 2760 gataaccaac gccttttgtc cactctagaa taatgaaagc tt 2802

    <210> 10 <211> 925 <212> PRT <213> Artificial Sequence <22O>

    <221> source <223> /note=Description of Artificial Sequence: synthetic polypeptide <400> 10

    Glu 1 Phe Val Asn Lys 5 Gin Phe Asn Tyr Lys Asp 10 Pro val Asn Gly 15 val Asp Ile Ala Tyr lie Lys lie Pro Asn Ala Gly Gin Met Gin Pro val 20 25 30 Lys Ala Phe Lys lie His Asn Lys Ile Trp val ile pro Glu Arg Asp 35 40 45 Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala 50 55 60 Lys Gin Val pro Val Ser Tyr Tyr Asp ser Thr Tyr Leu Ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly val Thr Lys Leu Phe Glu Arg

    85 90 95

    Page 14

    2015268754 16 Dec 2015

    lie P31 003a u-pc t.tx t Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser lie Val Arg 100 105 110 Gly Ile pro Phe Trp Gly Gly Ser Thr ile Asp Thr Glu Leu Lys val 115 120 125 lie Asp Thr Asn cys lie Asn val lie Gin Pro Asp Gly Ser Tyr Arg 130 135 140 Ser Glu Glu Leu Asn Leu Val ile lie Gly Pro Ser Ala Asp lie ile 145 150 155 160 Gin Phe Glu Cys Leu Ser Phe Gly Hi s Glu val Leu Asn Leu Thr Arg 165 170 175 Asn Gly Tyr Gly Ser Thr Gin Tyr Ile Arg Phe Ser Pro Asp Phe Thr 180 185 190 Phe Gly Phe Glu Glu ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly 195 200 205 Ala Gly Lys Phe Ala Thr Asp Pro Ala val Thr Leu Ala Hi s Glu Leu 210 215 220 lie His Ala Gly His Arg Leu Tyr Gly lie Ala Ile Asn Pro Asn Arg 225 230 235 240 val Phe Lys val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu 245 250 255 val Ser Phe Glu Glu Leu Arg Thr phe Gly Gly His Asp Ala Lys Phe 260 265 270 Ile Asp ser 275 Leu Gin Glu Asn Glu 280 phe Arg Leu Tyr Tyr 285 Tyr Asn Lys Phe Lys ASp lie Ala ser Thr Leu Asn Lys Ala Lys Ser Ile Val Gly 290 295 300 Thr Thr Ala ser Leu Gin Tyr Met Lys Asn val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu lie Tyr Thr Glu Asp Asn 340 345 350 Phe val Lys Phe Phe Lys val Leu Asn Arg Lys Thr Tyr Leu Asn Phe 355 360 365

    Page 15

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Asp Lys 370 Ala Val Phe Lys ile 375 Asn lie val Pro Lys 380 val Asn Tyr Thr Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe 385 390 395 400 Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu Lys 405 410 415 Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu cys Val Asp Asn 420 425 430 Asn Asn Asn Asn Asn Asn Asn Asn Asn Asp Asp Asp Asp Lys Asn Ser 435 440 445 Asp ser Glu Cys Pro Leu Ser Hi s Asp Gly Tyr cys Leu Hi s Asp Gly 450 455 460 Val Cys Met Tyr lie Glu Ala Leu Asp Lys Tyr Al a Cys Asn Cys Val 465 470 475 480 val Gly Tyr lie Gly Glu Arg cys Gln Tyr Arg Asp Leu Lys Trp Trp 485 490 495 Glu Leu Arg Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu 500 505 510 Gln Cys lie Lys val Asn Asn Trp Asp Leu Phe Phe ser Pro Ser Glu 515 520 525 Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu Ile Thr Ser Asp 530 535 540 Thr Asn lie Glu Ala Ala Glu Glu Asn lie Ser Leu Asp Leu lie Gln 545 550 555 560 Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro Glu Asn lie Ser 565 570 575 Ile Glu Asn Leu Ser Ser Asp Ile Ile Gly Gln Leu Glu Leu Met Pro 580 585 590 Asn lie Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu Leu Asp Lys Tyr 595 600 605 Thr Met Phe Hi s Tyr Leu Arg Ala Gln Glu Phe Glu Hi s Gly Lys Ser 610 615 620 Arg ile Ala Leu Thr Asn Ser val Asn Glu Ala Leu Leu Asn Pro ser 625 630 635 640

    Page 16

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    P31OO3au-pct.txt

    Arg val Tyr Thr Phe Phe Ser Ser Asp Tyr Val Lys Lys Val Asn Lys 645 650 655

    Ala Thr Glu Ala Ala Met Phe Leu Gly Trp val Glu Gln Leu val Tyr 660 665 670 Asp Phe Thr Asp Glu Thr ser Glu Val Ser Thr Thr Asp Lys lie Ala 675 680 685 Asp lie Thr Ile lie lie Pro Tyr ile Gly Pro Ala Leu Asn lie Gly 690 695 700 Asn Met Leu Tyr Lys Asp ASP Phe Val Gly Ala Leu Ile Phe Ser Gly 705 710 715 720 Ala val lie Leu Leu Glu Phe Ile Pro Glu Ile Ala ile Pro Val Leu 725 730 735 Gly Thr Phe Ala Leu val Ser Tyr Ile Ala Asn Lys val Leu Thr val 740 745 750 Gln Thr lie Asp Asn Ala Leu Ser Lys Arg Asn Glu Lys Trp Asp Glu 755 760 765 val Tyr Lys Tyr lie val Thr Asn Trp Leu Al a Lys Val Asn Thr Gln 770 775 780 lie Asp Leu lie Arg Lys Lys Met Lys Glu Ala Leu Glu Asn Gln Ala 785 790 795 800 Glu Ala Thr Lys Ala lie Ile Asn Tyr Gln Tyr Asn Gln Tyr Thr Glu 805 810 815 Glu Glu Lys Asn Asn Ile Asn Phe Asn Ile Asp Asp Leu Ser ser Lys 820 825 830 Leu Asn Glu Ser lie Asn Lys Ala Met lie Asn Ile Asn Lys Phe Leu 835 840 845 Asn Gln cys ser val ser Tyr Leu Met Asn Ser Met Ile Pro Tyr Gly 850 855 860 val Lys Arg Leu Glu Asp Phe Asp Ala ser Leu Lys Asp Ala Leu Leu 865 870 875 880 Lys Tyr ile Tyr Asp Asn Arg Gly Thr Leu lie Gly Gln val Asp Arg 885 890 895 Leu Lys Asp Lys val Asn Asn Thr Leu Ser Thr Asp lie Pro Phe Gln 900 905 910

    Page 17

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    P31003au-pct.txt

    Leu Ser Lys Tyr val Asp Asn Gin Arg Leu Leu ser Thr 915 920 925 <210> 11 <211> 864 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polypeptide <400> 11

    Glu 1 Phe Val Asn Lys Gin Phe Asn Tyr Lys Asp Pro val Asn Gly 15 val 5 10 Asp lie Ala Tyr lie Lys lie pro Asn Ala Gly Gin Met Gin Pro val 20 25 30 Lys Ala Phe Lys lie His Asn Lys lie Trp val lie Pro Glu Arg Asp 35 40 45 Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala 50 55 60 Lys Gin val pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly val Thr Lys Leu Phe Glu Arg 85 90 95 lie Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser lie Val Arg 100 105 110 Gly 'lie Pro Phe Trp Gly Gly Ser Thr lie Asp Thr Glu Leu Lys Val 115 120 125 lie Asp Thr Asn cys lie Asn Val lie Gin Pro Asp Gly Ser Tyr Arg 130 135 140 Ser Glu Glu Leu Asn Leu Val lie lie Gly Pro Ser Ala Asp lie lie 145 150 155 160 Gin Phe Glu cys Lys Ser Phe Gly Hi s Glu Val Leu Asn Leu Thr Arg 165 170 175 Asn Gly Tyr Gly Ser Thr Gin Tyr lie Arg Phe Ser pro Asp Phe Thr 180 185 190 Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly 195 200 205 Ala Gly Lys Phe Ala Thr Asp Pro Ala val Thr Leu Ala His Glu Leu

    page 18

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    P31OO3au-pct.txt

    210 215 220

    lie 225 His Ala Gly His Arg Leu Tyr Gly lie 230 Ala 235 lie Asn Pro Asn Arg 240 Val Phe Lys val Asn Thr Asn Ala Tyr Tyr Glu Met ser Gly Leu Glu 245 250 255 val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe 260 265 270 lie Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys 275 280 285 Phe Lys Asp lie Ala ser Thr Leu Asn Lys Ala Lys Ser lie val Gly 290 295 300 Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu Ser Glu Asp Thr ser Gly Lys Phe Ser val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu lie Tyr Thr Glu ASP Asn 340 345 350 Phe val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn Phe 355 360 365 Asp Lys Ala Val Phe Lys lie Asn lie val Pro Lys val Asn Tyr Thr 370 375 380 lie Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe 385 390 395 400 Asn Gly Gin Asn Thr Glu lie Asn Asn Met Asn Phe Thr Lys Leu Lys 405 410 415 Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys val Asp Gly 420 425 430 lie He Thr Ser Lys Thr Lys Ser Asp Asp Asp Asp Lys Asn Lys Ala 435 440 445 Leu Asn Leu Gin Cys lie Lys Val Asn Asn Trp Asp Leu Phe Phe Ser 450 455 460 Pro ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu lie 465 470 475 480 Thr ser Asp Thr Asn lie Glu Ala Ala Glu Glu Asn lie Ser Leu Asp

    Page 19

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    P31OO3au-pct.txt

    485 490 495 Leu lie Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro Glu 500 505 510 Asn lie Ser ile Glu Asn Leu Ser Ser Asp lie lie Gly Gln Leu Glu 515 520 525 Leu Met Pro Asn ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu Leu 530 535 540 Asp Lys Tyr Thr Met phe His Tyr Leu Arg Ala Gln Glu Phe Glu Hi s 545 550 555 560 Gly Lys Ser Arg ile Ala Leu Thr Asn ser Val Asn Glu Ala Leu Leu 565 570 575 Asn pro Ser Arg Val Tyr Thr Phe Phe Ser Ser Asp Tyr val Lys Lys 580 585 590 val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp val Glu Gln 595 600 605 Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu val ser Thr Thr Asp 610 615 620 Lys Ile Ala Asp Ile Thr lie ile lie Pro Tyr Ile Gly pro Al a Leu 625 630 635 640 Asn lie Gly Asn Met Leu Tyr Lys Asp Asp Phe val Gly Ala Leu Ile 645 650 655 Phe ser Gly Al a val ile Leu Leu Glu Phe Ile Pro Glu lie Al a lie 660 665 670 Pro val Leu Gly Thr Phe Ala Leu val ser Tyr lie Ala Asn Lys Val 675 680 685 Leu Thr Val Gln Thr ile Asp Asn Ala Leu Ser Lys Arg Asn Glu Lys 690 695 700 Trp Asp Glu val Tyr Lys Tyr ile val Thr Asn Trp Leu Ala Lys val 705 710 715 720 Asn Thr Gln lie Asp Leu lie Arg Lys Lys Met Lys Glu Ala Leu Glu 725 730 735 Asn Gln Ala Glu Ala Thr Lys Ala lie Ile Asn Tyr Gln Tyr Asn Gln 740 745 750 Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn lie Asp Asp Leu Pag ie 20

    2015268754 16 Dec 2015

    P31OO3au-pct.txt

    755

    760

    765

    Ser Ser Lys Leu Asn Glu Ser 775 Ile Asn Lys Ala Met 780 Ile Asn Ile Asn 770 Lys Phe Leu Asn Gin Cys Ser Val Ser Tyr Leu Met Asn Ser Met lie 785 790 795 800 Pro Tyr Gly val Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys Asp 805 810 815 Ala Leu Leu Lys Tyr ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly Gin 820 825 830 Val Asp Arg Leu Lys Asp Lys val Asn Asn Thr Leu Ser Thr Asp lie 835 840 845 Pro Phe Gin Leu Ser Lys Tyr val Asp Asn Gin Arg Leu Leu Ser Thr 850 855 860

    <210> 12 <211> 869 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 12

    Pro 1 val Thr ile Asn 5 Asn Phe Asn Tyr Asn Asp 10 Pro lie Asp Asn 15 Asn Asn lie lie Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg Tyr 20 25 30 Tyr Lys Ala Phe Lys Ile Thr Asp Arg ile Trp Ile lie Pro Glu Arg 35 40 45 Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly Ile 50 55 60 Phe Asn Arg Asp val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn Thr 65 70 75 80 Asn Asp Lys Lys Asn lie Phe Leu Gin Thr Met lie Lys Leu Phe Asn 85 90 95 Arg Ile Lys ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met lie lie 100 105 110 Asn Gly Ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu Phe

    115 120 125

    Page 21

    P31003au-pct.txt

    2015268754 16 Dec 2015

    Asn Thr Asn lie Ala ser Val Thr Val Asn Lys Leu 140 lie Ser Asn Pro 130 135 Gly Glu val Glu Arg Lys Lys Gly lie Phe Ala Asn Leu lie lie Phe 145 150 155 160 Gly Pro Gly Pro val Leu Asn Glu Asn Glu Thr lie Asp He Gly lie 165 170 175 Gin Asn Hi s Phe Ala Ser Arg Glu Gly Phe Gly Gly lie Met Gin Met 180 185 190 Lys Phe Cys Pro Glu Tyr val ser Val Phe Asn Asn val Gin Glu Asn 195 200 205 Lys Gly Ala Ser lie Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro Ala 210 215 220 Leu lie Leu Met Hi s Glu Leu lie Hi s val Leu Hi s Gly Leu Tyr Gly 225 230 235 240 lie Lys val Asp Asp Leu pro lie val Pro Asn Glu Lys Lys Phe Phe 245 250 255 Met Gin Ser Thr Asp Ala lie Gin Ala Glu Glu Leu Tyr Thr Phe Gly 260 265 270 Gly Gin Asp Pro Ser lie lie Thr Pro Ser Thr Asp Lys ser lie Tyr 275 280 285 Asp Lys val Leu Gin Asn Phe Arg Gly lie Val Asp Arg Leu Asn Lys 290 295 300 val Leu val Cys lie Ser ASp Pro Asn lie Asn lie Asn lie Tyr Lys 305 310 315 320 Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly Lys 325 330 335 Tyr Ser lie Asp val Glu ser Phe Asp Lys Leu Tyr Lys ser Leu Met 340 345 350 Phe Gly Phe Thr Glu Thr Asn lie Ala Glu Asn Tyr Lys lie Lys Thr 355 360 365 Arg Ala Ser Tyr Phe ser Asp Ser Leu Pro Pro val Lys lie Lys Asn 370 375 380 Leu Leu Asp Asn Glu lie Tyr Thr lie Glu Glu Gly Phe Asn lie ser

    385 390 395 400

    Page 22

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    P31OO3au-pct.txt

    Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gin Asn Lys Ala lie Asn 405 410 415 Lys Gin Ala Tyr Glu Glu lie Ser Lys Glu His Leu Ala Val Tyr Lys 420 425 430 lie Gin Met Cys val Asp Glu Glu Lys Leu Tyr Asp Asp Asp Asp Lys 435 440 445 Asp Arg Trp Gly Ser Ser Leu Gin cys He Asp Val Asp Asn Glu Asp 450 455 460 Leu Phe Phe lie Ala Asp Lys Asn ser Phe Ser ASP Asp Leu Ser Lys 465 470 475 480 Asn Glu Arg lie Glu Tyr Asn Thr Gin Ser Asn Tyr lie Glu Asn Asp 485 490 495 Phe Pro lie Asn Glu Leu lie Leu Asp Thr Asp Leu lie Ser Lys lie 500 505 510 Glu Leu Pro Ser Glu Asn Thr Glu Ser Leu Thr Asp Phe Asn val Asp 515 520 525 val Pro val Tyr Glu Lys Gin Pro Ala lie Lys Lys lie Phe Thr Asp 530 535 540 Glu Asn Thr lie Phe Gin Tyr Leu Tyr Ser Gin Thr Phe Pro Leu Asp 545 550 555 560 lie Arg Asp lie Ser Leu Thr Ser Ser Phe Asp Asp Ala Leu Leu Phe 565 570 575 Ser Asn Lys Val Tyr Ser Phe Phe Ser Met ASp Tyr lie Lys Thr Ala 580 585 590 Asn Lys val Val Glu Ala Gly Leu Phe Ala Gly Trp Val Lys Gin lie 595 600 605 val Asn Asp Phe val lie Glu Ala Asn Lys Ser Asn Thr Met Asp Ala 610 615 620 lie Ala Asp lie ser Leu lie val Pro Tyr lie Gly Leu Ala Leu Asn 625 630 635 640 Val Gly Asn Glu Thr Ala Lys Gly Asn Phe Glu Asn Ala Phe Glu lie 645 650 655 Ala Gly Ala Ser lie Leu Leu Glu Phe lie Pro Glu Leu Leu He Pro 660 665 670

    Page 23

    P31003au-pct.txt

    2015268754 16 Dec 2015

    val Val Gly Ala Phe Leu Leu Glu Ser Tyr lie Asp Asn 685 Lys Asn Lys 675 680 Ile lie Lys Thr ile Asp Asn Ala Leu Thr Lys Arg Asn Glu Lys Trp 690 695 700 ser Asp Met Tyr Gly Leu Ile Val Ala Gln Trp Leu Ser Thr val Asn 705 710 715 720 Thr Gln phe Tyr Thr Ile Lys Glu Gly Met Tyr Lys Ala Leu Asn Tyr 725 730 735 Gln Ala Gln Al a Leu Glu Glu ile Ile Lys Tyr Arg Tyr Asn lie Tyr 740 745 750 Ser Glu Lys Glu Lys ser Asn Ile Asn ile Asp Phe Asn Asp Ile Asn 755 760 765 Ser Lys Leu Asn Glu Gly lie Asn Gln Ala lie Asp Asn Ile Asn Asn 770 775 780 Phe lie Asn Gly cys Ser val Ser Tyr Leu Met Lys Lys Met lie Pro 785 790 795 800 Leu Ala val Gl u Lys Leu Leu Asp Phe Asp Asn Thr Leu Lys Lys Asn 805 810 815 Leu Leu Asn Tyr Ile Asp Glu Asn Lys Leu Tyr Leu ile Gly ser Al a 820 825 830 Glu Tyr Glu Lys Ser Lys Val Asn Lys Tyr Leu Lys Thr ile Met Pro 835 840 845 phe Asp Leu Ser lie Tyr Thr Asn Asp Thr Ile Leu Ile Glu Met Phe 850 855 860

    Asn Lys Tyr Asn Ser 865 <210> 13 <211> 865 <212> PRT <213> Artificial sequence <22O>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polypeptide <400> 13

    Pro Ile Thr lie Asn Asn Phe Asn Tyr Ser Asp Pro val Asp Asn Lys 15 10 15

    Page 24

    P31003au-pct.txt

    2015268754 16 Dec 2015

    Asn Ile Leu Tyr Leu 20 Asp Thr His Leu Asn Thr Leu Ala Asn Glu Pro 25 30 Glu Lys Ala Phe Arg Ile Thr Gly Asn Ile Trp val lie Pro Asp Arg 35 40 45 Phe ser Arg Asn ser Asn Pro Asn Leu Asn Lys Pro Pro Arg val Thr 50 55 60 Ser Pro Lys ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp Ser 65 70 75 80 Asp Lys Asp Thr Phe Leu Lys Glu lie Ile Lys Leu Phe Lys Arg Ile 85 90 95 Asn Ser Arg Glu Ile Gly Glu Glu Leu lie Tyr Arg Leu ser Thr Asp 100 105 110 lie Pro Phe Pro Gly Asn Asn Asn Thr Pro lie Asn Thr Phe Asp Phe 115 120 125 Asp val ASp Phe Asn ser Val Asp Val Lys Thr Arg Gin Gly Asn Asn 130 135 140 Trp val Lys Thr Gly Ser Ile Asn Pro Ser val lie lie Thr Gly Pro 145 150 155 160 Arg Glu Asn lie lie Asp Pro Glu Thr ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Ala Ala Gin Glu Gly Phe Gly Ala Leu Ser lie Ile Ser 180 185 190 Ile ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp Val 195 200 205 Gly Glu Gly Arg Phe Ser Lys Ser Glu phe cys Met ASP Pro lie Leu 210 215 220 Ile Leu Met Hi s Glu Leu Asn Hi s Ala Met His Asn Leu Tyr Gly lie 225 230 235 240 Ala lie Pro Asn Asp Gin Thr lie Ser Ser val Thr Ser Asn lie Phe 245 250 255 Tyr ser Gin Tyr Asn val Lys Leu Glu Tyr Ala Glu lie Tyr Ala Phe 260 265 270 Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr Phe 275 280 285

    Page 25

    P31003au-pct.txt

    2015268754 16 Dec 2015

    Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser lie Ala Lys Arg Leu Asn 290 295 300 Ser lie Thr Thr Ala Asn Pro ser Ser Phe Asn Lys Tyr lie Gly Glu 305 310 315 320 Tyr Lys Gin Lys Leu lie Arg Lys Tyr Arg Phe val val Glu Ser Ser 325 330 335 Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn Glu 340 345 350 Leu Thr Gin lie Phe Thr Glu Phe Asn Tyr Al a Lys lie Tyr Asn val 355 360 365 Gin Asn Arg Lys lie Tyr Leu Ser Asn val Tyr Thr pro Val Thr Ala 370 375 380 Asn lie Leu ASp Asp Asn Val Tyr Asp lie Gin Asn Gly Phe Asn lie 385 390 395 400 Pro Lys Ser Asn Leu Asn val Leu Phe Met Gly Gin Asn Leu ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe Cys val Asp Ala lie Asp Gly Arg Ser Leu Tyr Asn Lys 435 440 445 Thr Leu Gin cys Arg Glu Leu Leu val Lys Asn Thr ASp Leu Pro Phe 450 455 460 lie Gly Asp lie Ser Asp Val Lys Thr Asp lie Phe Leu Arg Lys Asp 465 470 475 480 lie Asn Glu Glu Thr Glu Val lie Tyr Tyr Pro Asp Asn Val Ser Val 485 490 495 Asp Gin Val lie Leu Ser Lys Asn Thr Ser Glu Hi s Gly Gin Leu Asp 500 505 510 Leu Leu Tyr Pro Ser He Asp ser Glu Ser Glu lie Leu Pro Gly Glu 515 520 525 Asn Gin Val Phe Tyr Asp Asn Arg Thr Gin Asn val Asp Tyr Leu Asn 530 535 540 ser Tyr Tyr Tyr Leu Glu ser Gin Lys Leu Ser Asp Asn val Glu Asp 545 550 555 560

    Page 26

    P31003au-pct.txt

    2015268754 16 Dec 2015

    Phe Thr Phe Thr Arg Ser Ile Glu Glu Ala Leu Asp Asn Ser Ala 575 Lys 565 570 val Tyr Thr Tyr Phe pro Thr Leu Ala Asn Lys Val Asn Ala Gly val 580 585 590 Gin Gly Gly Leu Phe Leu Met Trp Ala Asn Asp val val Glu Asp Phe 595 600 605 Thr Thr Asn lie Leu Arg Lys Asp Thr Leu ASP Lys Ile ser Asp Val 610 615 620 Ser Ala Ile lie Pro Tyr Ile Gly Pro Ala Leu Asn Ile Ser Asn Ser 625 630 635 640 val Arg Arg Gly Asn Phe Thr Glu Ala Phe Ala Val Thr Gly Val Thr 645 650 655 lie Leu Leu Glu Ala Phe Pro Glu Phe Thr lie Pro Ala Leu Gly Ala 660 665 670 Phe Val lie Tyr Ser Lys Val Gin Glu Arg Asn Glu Ile Ile Lys Thr 675 680 685 lie Asp Asn Cys Leu Glu Gin Arg lie Lys Arg Trp Lys Asp Ser Tyr 690 695 700 Glu Trp Met Met Gly Thr Trp Leu ser Arg lie lie Thr Gin Phe Asn 705 710 715 720 Asn lie Ser Tyr Gin Met Tyr Asp Ser Leu Asn Tyr Gin Ala Gly Ala 725 730 735 Ile Lys Ala Lys Ile Asp Leu Glu Tyr Lys Lys Tyr Ser Gly Ser Asp 740 745 750 Lys Glu Asn Ile Lys ser Gin Val Glu Asn Leu Lys Asn ser Leu Asp 755 760 765 val Lys lie Ser Glu Ala Met Asn Asn lie Asn Lys Phe lie Arg Glu 770 775 780 Cys ser val Thr Tyr Leu Phe Lys Asn Met Leu Pro Lys val lie Asp 785 790 795 800 Glu Leu Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu lie Asn Leu 805 810 815 Ile Asp Ser His Asn lie Ile Leu Val Gly Glu val Asp Lys Leu Lys 820 825 830

    Page 27

    2015268754 16 Dec 2015

    P31003au-pct.txt

    Ala Lys Val Asn Asn Ser Phe Gin Asn Thr Ile Pro Phe Asn lie Phe 835 840 845

    Ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp lie lie Asn Glu Tyr phe 850 855 860

    Asn

    865 <210> 14 <211> 871 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial sequence: synthetic polypeptide

    <400> 14 Lys 5 Asp Phe Asn Tyr Ser Asp Pro 10 Val Asn Asp 15 Asn Thr 1 Trp Pro val Asp lie Leu Tyr Leu Arg lie Pro Gin Asn Lys Leu lie Thr Thr Pro 20 25 30 Val Lys Ala Phe Met He Thr Gin Asn lie T rp val lie Pro Glu Arg 35 40 45 Phe ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gin Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 Gin Lys Asp Thr Phe Leu Lys Gly He lie Lys Leu Phe Lys Arg lie 85 90 95 Asn Glu Arg Asp lie Gly Lys Lys Leu Ile Asn Tyr Leu val Val Gly 100 105 110 Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg Hi s Thr Thr Asn Ile Ala val Glu Lys Phe Glu Asn Gly Ser 130 135 140 Trp Lys Val Thr Asn lie Ile Thr Pro Ser val Leu Ile Phe Gly Pro 145 150 155 160 Leu Pro Asn Ile Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gin Gly Gin 165 170 175

    Page 28

    2015268754 16 Dec 2015

    P31 003a u-pc t.tx t Gin Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu Lys 180 185 190 val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr ser Asn Gin 195 200 205 Ser Ser Al a val Leu Gly Lys Ser lie Phe cys Met ASp Pro val lie 210 215 220 Ala Leu Met Hi s Glu Leu Thr His Ser Leu Hi s Gin Leu Tyr Gly lie 225 230 235 240 Asn lie Pro ser Asp Lys Arg Ile Arg Pro Gin val Ser Glu Gly Phe 245 250 255 Phe ser Gin Asp Gly Pro Asn Val Gin Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu ASp Val Glu He Ile Pro Gin Ile Glu Arg Ser Gin Leu 275 280 285 Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu Asn 290 295 300 Asn lie Asn Lys Thr Ile Pro Ser Ser Trp lie Ser Asn lie Asp Lys 305 310 315 320 Tyr Lys Lys lie Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Gly Asn Phe val val Asn lie Asp Lys Phe Asn Ser Leu Tyr ser Asp 340 345 350 Leu Thr Asn Val Met Ser Glu val Val Tyr Ser Ser Gin Tyr Asn Val 355 360 365 Lys Asn Arg Thr Hi s Tyr Phe Ser Arg His Tyr Leu Pro val Phe Ala 370 375 380 Asn lie Leu Asp Asp Asn lie Tyr Thr lie Arg Asp Gly Phe Asn Leu 385 390 395 400 Thr Asn Lys Gly Phe Asn lie Glu Asn Ser Gly Gin Asn lie Glu Arg 405 410 415 Asn Pro Ala Leu Gin Lys Leu Ser Ser Glu Ser val val Asp Leu Phe 420 425 430 Thr Lys val Cys val Asp Lys ser Glu Glu Lys Leu Tyr Asp Asp Asp 435 440 445

    Page 29

    P31OO3au-pct.txt Asp Lys Asp Arg Trp Gly ser ser Leu Gln Cys ile Lys val Lys Asn o 450 455 460 CN o /1 Λ Asn Arg Leu Pro Tyr val Ala Asp Lys Asp Ser lie ser Gln Glu lie QJ Q 465 470 475 480 kO Phe Glu Asn Lys lie Ile Thr Asp Glu Thr Asn Val Gln Asn Tyr ser 485 490 495 Vj- Asp Lys Phe Ser Leu Asp Glu ser lie Leu Asp Gly Gln val Pro lie in 500 505 510 o oo kO Asn Pro Glu lie val Asp pro Leu Leu Pro Asn val Asn Met Glu Pro CN 515 520 525 on o Leu Asn Leu Pro Gly Glu Glu lie Val Phe Tyr Asp ASp Ile Thr Lys CN 530 535 540

    Tyr Val Asp Tyr 545 Leu Asn ser Tyr Tyr Tyr Leu Glu Ser Gln Lys Leu 560 550 555 Ser Asn Asn val Glu Asn Ile Thr Leu Thr Thr Ser val Glu Glu Ala 565 570 575 Leu Gly Tyr Ser Asn Lys Ile Tyr Thr Phe Leu Pro ser Leu Ala Glu 580 585 590 Lys val Asn Lys Gly val Gln Ala Gly Leu Phe Leu Asn Trp Ala Asn 595 600 605 Glu val Val Glu Asp Phe Thr Thr Asn lie Met Lys Lys Asp Thr Leu 610 615 620 Asp Lys ile Ser Asp Val Ser Val lie lie Pro Tyr lie Gly Pro Ala 625 630 635 640 Leu Asn Ile Gly Asn Ser Ala Leu Arg Gly Asn Phe Asn Gln Ala Phe 645 650 655 Ala Thr Ala Gly Val Ala Phe Leu Leu Glu Gly Phe Pro Glu Phe Thr 660 665 670 Ile Pro Ala Leu Gly Val Phe Thr Phe Tyr Ser Ser lie Gln Glu Arg 675 680 685 Glu Lys Ile Ile Lys Thr Ile Glu Asn cys Leu Glu Gln Arg Val Lys 690 695 700 Arg Trp Lys Asp ser Tyr Gln Trp Met val Ser Asn Trp Leu Ser Arg 705 710 715 720

    Page 30

    2015268754 16 Dec 2015

    lie Thr Thr P31003au-pct. txt Gin Phe 725 Asn His lie Asn Tyr 730 Gin Met Tyr Asp Ser 735 Leu Ser Tyr Gin Ala Asp Ala lie Lys Ala Lys Ile ASp Leu Glu Tyr Lys 740 745 750 Lys Tyr Ser Gly Ser Asp Lys Glu Asn lie Lys ser Gin Val Glu Asn 755 760 765 Leu Lys Asn Ser Leu Asp Val Lys Ile Ser Glu Ala Met Asn Asn He 770 775 780 Asn Lys Phe Ile Arg Glu Cys Ser val Thr Tyr Leu Phe Lys Asn Met 785 790 795 800 Leu Pro Lys val ile Asp Glu Leu Asn Lys Phe Asp Leu Arg Thr Lys 805 810 815 Thr Glu Leu ile Asn Leu lie Asp Ser Hi s Asn Ile Ile Leu Val Gly 820 825 830 Glu Val Asp Arg Leu Lys Ala Lys Val Asn Glu Ser Phe Glu Asn Thr 835 840 845 Met Pro Phe Asn lie Phe Ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp 850 855 860 lie lie Asn Glu Tyr Phe Asn 865 870

    <210> 15 <211> 10 <212> PRT <213> Artificial sequence <22O>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic peptide <400> 15

    Glu His Trp Ser Tyr Gly Cys Arg Pro Gly

    1 5 10 <210> 16 <211> 920 <212> PRT <213> Artificial Sequence <22O>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 16

    Pro Val Thr lie Asn Asn Phe Asn Tyr Asn Asp Pro Ile Asp Asn Asn 15 10 15

    Page 31

    P31OO3au-pct.txt υη

    Ο (Μ

    Ο

    Ω ’tjοο (Μ ο

    (Μ

    Asn ile lie Met Met 20 Glu Pro Pro Phe Ala Arg 25 Gly Thr Gly Arg Tyr 30 Tyr Lys Ala Phe Lys lie Thr Asp Arg lie Trp Ile Ile Pro Glu Arg 35 40 45 Tyr Thr Phe Gly Tyr Lys Pro Glu Asp phe Asn Lys Ser Ser Gly ile 50 55 60 phe Asn Arg Asp val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn Thr 65 70 75 80 Asn Asp Lys Lys Asn lie Phe Leu Gln Thr Met lie Lys Leu Phe Asn 85 90 95 Arg Ile Lys ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met lie lie 100 105 110 Asn Gly lie Pro Tyr Leu Gly Asp Arg Arg val Pro Leu Glu Glu Phe 115 120 125 Asn Thr Asn lie Ala ser Val Thr val Asn Lys Leu Ile Ser Asn Pro 130 135 140 Gly Glu val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu lie Ile Phe 145 150 155 160 Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr lie Asp Ile Gly Ile 165 170 175 Gln Asn Hi s Phe Ala Ser Arg Glu Gly Phe Gly Gly lie Met Gln Met 180 185 190 Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gln Glu Asn 195 200 205 Lys Gly Ala Ser ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro Ala 210 215 220 Leu ile Leu Met His Glu Leu lie Hi s val Leu His Gly Leu Tyr Gly 225 230 235 240 Ile Lys val Asp Asp Leu Pro Ile Val Pro Asn Glu Lys Lys Phe Phe 245 250 255 Met Gln Ser Thr Asp Ala lie Gln Ala Glu Glu Leu Tyr Thr Phe Gly 260 265 270 Gly Gln ASp Pro ser lie ile Thr Pro Ser Thr Asp Lys Ser lie Tyr 275 280 285

    Page 32

    2015268754 16 Dec 2015

    P31OO3au-pct.txt

    Asp Lys val Leu Gln Asn Phe 295 Arg Gly Ile Val Asp 300 Arg Leu Asn Lys 290 val Leu val Cys lie Ser Asp Pro Asn lie Asn lie Asn Ile Tyr Lys 305 310 315 320 Asn Lys Phe Lys Asp Lys Tyr Lys phe Val Glu ASp Ser Glu Gly Lys 325 330 335 Tyr Ser ile Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu Met 340 345 350 Phe Gly Phe Thr Glu Thr Asn Ile Ala Glu Asn Tyr Lys Ile Lys Thr 355 360 365 Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro val Lys Ile Lys Asn 370 375 380 Leu Leu Asp Asn Glu lie Tyr Thr Ile Glu Glu Gly Phe Asn lie Ser 385 390 395 400 Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gln Asn Lys Ala Ile Asn 405 410 415 Lys Gln Ala Tyr Glu Glu Ile ser Lys Glu Hi s Leu Al a Val Tyr Lys 420 425 430 lie Gln Met Cys val Asp Gly Ile lie Thr Ser Lys Thr Lys Ser Asp 435 440 445 Asp Asp ASp Lys Asn Lys Ala Leu Asn Leu Gln Cys Ile ASp val Asp 450 455 460 Asn Glu ASp Leu Phe Phe Ile Ala Asp Lys Asn ser Phe ser Asp Asp 465 470 475 480 Leu Ser Lys Asn Glu Arg Ile Glu Tyr Asn Thr Gln Ser Asn Tyr Ile 485 490 495 Glu Asn Asp Phe Pro Ile Asn Glu Leu Ile Leu Asp Thr Asp Leu Ile 500 505 510 ser Lys ile Glu Leu Pro ser Glu Asn Thr Glu Ser Leu Thr Asp Phe 515 520 525 Asn val 530 ASp val Pro val Tyr 535 Glu Lys Gln Pro Ala 540 Ile Lys Lys Ile Phe Thr Asp Glu Asn Thr Ile Phe Gln Tyr Leu Tyr Ser Gln Thr Phe 545 550 555 560

    Page 33

    P31003au-pct.txt

    Ο CM Pro Leu Asp lie Arg Asp lie Ser Leu Thr Ser ser Phe Asp ASp Ala 565 570 575 Ο <υ Ω Leu Leu Phe Ser Asn Lys Val Tyr ser Phe Phe ser Met Asp Tyr lie kO 580 585 590 r—Η Lys Thr Ala Asn Lys val val Glu Ala Gly Leu Phe Ala Gly Trp val 595 600 605 τΤ ο Lys Gin lie val Asn Asp Phe val lie Glu Ala Asn Lys ser Asn Thr οο 610 615 620 <ο CM <ΖΊ Met ASp Lys lie Ala Asp lie ser Leu lie Val Pro Tyr He Gly Leu Η 625 630 635 640 Ο CM Ala Leu Asn val Gly Asn Glu Thr Ala Lys Gly Asn Phe Glu Asn Al a 645 650 655 Phe Glu lie Ala Gly Ala ser lie Leu Leu Glu Phe lie pro Glu Leu 660 665 670 Leu lie Pro Val val Gly Ala Phe Leu Leu Glu ser Tyr lie Asp Asn 675 680 685 Lys Asn Lys lie lie Lys Thr lie Asp Asn Ala Leu Thr Lys Arg Asn 690 695 700 Glu Lys Trp Ser Asp Met Tyr Gl y Leu lie val Ala Gin Trp Leu Ser 705 710 715 720 Thr Val Asn Thr Gin Phe Tyr Thr lie Lys Glu Gly Met Tyr Lys Ala 725 730 735 Leu Asn Tyr Gin Ala Gin Ala Leu Glu Glu lie lie Lys Tyr Arg Tyr 740 745 750 Asn lie Tyr Ser Glu Lys Glu Lys Ser Asn lie Asn lie ASp Phe Asn 755 760 765 Asp lie Asn Ser Lys Leu Asn Glu Gly lie Asn Gin Ala lie Asp Asn 770 775 780 lie Asn Asn Phe lie Asn Gly Cys Ser val Ser Tyr Leu Met Lys Lys 785 790 795 800 Met lie Pro Leu Ala Val Glu Lys Leu Leu Asp Phe Asp Asn Thr Leu 805 810 815 Lys Lys Asn Leu Leu Asn Tyr lie Asp Glu Asn Lys Leu Tyr Leu lie 820 825 830

    Page 34

    2015268754 16 Dec 2015

    P31003au-pct.txt

    Gly Ser Ala 835 Glu Tyr Glu Lys Ser Lys val Asn Lys Tyr Leu Lys Thr 840 845 lie Met Pro Phe Asp Leu ser lie Tyr Thr Asn Asp Thr lie Leu lie 850 855 860 Glu Met Phe Asn Lys Tyr Asn Ser Leu Glu Gly Gly Gly Gly Ser Gly 865 870 875 880 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Asp ser Ala Asn Ser 885 890 895 Asn Pro Ala Met Ala Pro Arg Glu Arg Lys Ala Gly Cys Lys Asn Phe 900 905 910

    Phe Trp Lys Thr Phe Thr ser cys 915 920 <210> 17 <211> 908 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: synthetic polypeptide <400> 17

    Glu Phe 1 Val Asn Lys 5 Gin Phe Asn Tyr Lys Asp Pro val Asn Gly val 10 15 Asp lie Ala Tyr lie Lys lie pro Asn Ala Gly Gin Met Gin Pro val 20 25 30 Lys Ala phe Lys lie Hi s Asn Lys lie Trp val lie Pro Glu Arg Asp 35 40 45 Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro pro Pro Glu Ala 50 55 60 Lys Gin val Pro val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu Phe Glu Arg 85 90 95 lie Tyr ser Thr Asp Leu Gly Arg Met Leu Leu Thr ser He val Arg 100 105 110 Gly lie Pro Phe Trp Gly Gly ser Thr lie Asp Thr Glu Leu Lys val

    115 120 125

    Page 35

    P31OO3au-pct.txt

    IT)

    S¹¹⁶ CH Asp Thr Asn Cys lie Asn val lie Gin Pro Asp Gly Ser Tyr Arg 130 Glu Glu 135 val lie 140 Asp He He 160 Q CD Ω Ser 145 Leu Asn Leu 150 lie Gly Pro 155 Ser Ala kO Gin Phe Glu Cys Leu Ser Phe Gly His Glu Val Leu Asn Leu Thr Arg 165 170 175 Tf IT) Asn Gly Tyr Gly Ser Thr Gin Tyr lie Arg Phe ser Pro Asp Phe Thr 180 185 190 oo <© CH IT) Phe Gly phe Glu Glu Ser Leu Glu val Asp Thr Asn Pro Leu Leu Gly 195 200 205 o CH Ala Gly Lys Phe Ala Thr Asp pro Ala Val Thr Leu Al a Hi s Glu Leu 210 215 220 lie Hi s Ala Gly Hi s Arg Leu Tyr Gly lie Ala lie Asn Pro Asn Arg 225 230 235 240 Val Phe Lys Val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu 245 250 255 Val Ser phe Glu Glu Leu Arg Thr Phe Gly Gly Hi s Asp Ala Lys Phe 260 265 270 lie Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys 275 280 285 Phe Lys Asp lie Ala Ser Thr Leu Asn Lys Ala Lys ser lie val Gly 290 295 300 Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu Ser Glu Asp Thr ser Gly Lys Phe Ser val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu lie Tyr Thr Gl u Asp Asn 340 345 350 Phe val Lys Phe Phe Lys val Leu Asn Arg Lys Thr Tyr Leu Asn Phe 355 360 365 Asp Lys Ala val Phe Lys lie Asn lie Val pro Lys Val Asn Tyr Thr 370 375 380 lie Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn phe 385 390 395 400

    Page 36

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    P31OO3au-pct.txt

    Asn Gly Gln Asn Thr Glu lie Asn Asn Met Asn Phe Thr Lys Leu Lys 405 410 415 Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Asp Gly 420 425 430 lie lie Thr ser Lys Thr Lys ser Asp Asp Asp Asp Lys Ser Ala Asn 435 440 445 ser Asn Pro Al a Met Ala Pro Arg Glu Arg Lys Ala Gly cys Lys Asn 450 455 460 Phe Phe Trp Lys Thr Phe Thr Ser cys Ala Leu Ala Gly Gly Gly Gly 465 470 475 480 ser Gly Gly Gly Gly ser Gly Gly Gly Gly Ser Ala Leu Val Leu Gln 485 490 495 cys lie Lys val Asn Asn Trp Asp Leu Phe Phe Ser Pro Ser Glu Asp 500 505 510 Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu Ile Thr ser Asp Thr 515 520 525 Asn lie Glu Ala Ala Glu Glu Asn lie Ser Leu Asp Leu lie Gln Gln 530 535 540 Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro Glu Asn lie ser lie 545 550 555 560 Glu Asn Leu ser Ser Asp Ile ile Gly Gln Leu Glu Leu Met pro Asn 565 570 575 ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu Leu Asp Lys Tyr Thr 580 585 590 Met Phe His Tyr Leu Arg Ala Gln Glu Phe Glu Hi s Gly Lys Ser Arg 595 600 605 lie Ala Leu Thr Asn Ser Val Asn Glu Ala Leu Leu Asn Pro ser Arg 610 615 620 val Tyr Thr Phe Phe Ser Ser Asp Tyr val Lys Lys val Asn Lys Ala 625 630 635 640 Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu Gln Leu val Tyr Asp 645 650 655 Phe Thr Asp Glu Thr Ser Glu val Ser Thr Thr Asp Lys Ile Ala Asp 660 665 670

    Page 37

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    P31003au-pct.txt

    ile Thr lie 675 ile lie Pro Tyr lie Gly Pro Ala 680 Leu Asn 685 lie Gly Asn Met Leu Tyr Lys Asp ASp Phe Val Gly Ala Leu lie Phe Ser Gly Ala 690 695 700 val ile Leu Leu Glu Phe lie Pro Glu lie Ala Ile Pro Val Leu Gly 705 710 715 720 Thr Phe Ala Leu Val Ser Tyr He Ala Asn Lys val Leu Thr val Gin 725 730 735 Thr Ile Asp Asn Ala Leu ser Lys Arg Asn Glu Lys Trp Asp Glu Val 740 745 750 Tyr Lys Tyr Ile Val Thr Asn Trp Leu Ala Lys Val Asn Thr Gin Ile 755 760 765 Asp Leu Ile Arg Lys Lys Met Lys Glu Ala Leu Glu Asn Gin Ala Glu 770 775 780 Ala Thr Lys Al a lie lie Asn Tyr Gin Tyr Asn Gin Tyr Thr Glu Glu 785 790 795 800 Glu Lys Asn Asn lie Asn Phe Asn lie Asp ASp Leu Ser Ser Lys Leu 805 810 815 Asn Glu ser Ile Asn Lys Ala Met lie Asn lie Asn Lys Phe Leu Asn 820 825 830 Gin Cys ser Val Ser Tyr Leu Met Asn Ser Met Ile Pro Tyr Gly val 835 840 845 Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys Asp Ala Leu Leu Lys 850 855 860 Tyr Ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly Gin val Asp Arg Leu 865 870 875 880 Lys Asp Lys Val Asn Asn Thr Leu Ser Thr Asp Ile Pro Phe Gin Leu 885 890 895 Ser Lys Tyr Val Asp Asn Gin Arg Leu Leu Ser Thr 900 905

    <210> 18 <211> 944 <212> PRT <213> Artificial Sequence <22O>

    <221> source

    Page 38

    2015268754 16 Dec 2015

    P31OO3au-pct.txt <223> /note=Description of Artificial sequence: Synthetic polypeptide <400> 18

    Thr Trp Pro val 1 Lys 5 Asp Phe Asn Tyr Ser Asp 10 Pro val Asn Asp 15 Asn ASp lie Leu Tyr Leu Arg lie Pro Gin Asn Lys Leu lie Thr Thr Pro 20 25 30 val Lys Ala Phe Met lie Thr Gin Asn lie Trp val lie Pro Glu Arg 35 40 45 Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gin Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 Gin Lys Asp Thr Phe Leu Lys Gly lie lie Lys Leu Phe Lys Arg lie 85 90 95 Asn Glu Arg Asp lie Gly Lys Lys Leu lie Asn Tyr Leu Val val Gly 100 105 110 Ser Pro Phe Met Gly ASp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg Hi s Thr Thr Asn He Ala val Glu Lys Phe Glu Asn Gly ser 130 135 140 Trp Lys Val Thr Asn lie lie Thr Pro Ser val Leu lie Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gin Gly Gin 165 170 175 Gin Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser lie Leu Lys 180 185 190 Val Ala Pro Glu phe Leu Leu Thr Phe Ser Asp val Thr ser Asn Gin 195 200 205 Ser ser Ala val Leu Gly Lys Ser lie Phe Cys Met Asp Pro val lie 210 215 220 Ala Leu Met Hi s Glu Leu Thr His Ser Leu Hi s Gin Leu Tyr Gly lie 225 230 235 240 Asn lie Pro Ser Asp Lys Arg lie Arg pro Gin Val Ser Glu Gly Phe

    245 250 255

    Page 39

    ιζχ P3K D03ai j-pct.txt *4 J ³he Ser Gin Asp Gly Pro Asn val Gin Phe Glu Glu Leu Tyr Thr Phe o 260 265 270 CH O Gly Gly Leu Asp Val Glu lie lie Pro Gin lie Glu Arg Ser Gin Leu <D Q 275 280 285 Arg Glu Lys Ala Leu Gly Hi s Tyr Lys Asp lie Ala Lys Arg Leu Asn 290 295 300 Asn lie Asn Lys Thr lie Pro Ser Ser Trp lie Ser Asn lie Asp Lys XI in 305 310 315 320 o oo <O Tyr Lys Lys lie Phe ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr CH 325 330 335 m o Gly Asn Phe Val Val Asn lie Asp Lys Phe Asn ser Leu Tyr Ser Asp <N 340 345 350 Leu Thr Asn Val Met Ser Glu val Val Tyr ser ser Gin Tyr Asn Val 355 360 365 Lys Asn Arg Thr His Tyr Phe ser Arg Hi s Tyr Leu Pro val Phe Ala 370 375 380 Asn lie Leu Asp Asp Asn He Tyr Thr lie Arg Asp Gly Phe Asn Leu 385 390 395 400 Thr Asn Lys Gly Phe Asn lie Glu Asn ser Gly Gin Asn lie Glu Arg 405 410 415 Asn Pro Ala Leu Gin Lys Leu Ser Ser Glu Ser Val val Asp Leu Phe 420 425 430 Thr Lys Val cys val Asp Lys Ser Glu Glu Lys Leu Tyr Asp Asp Asp 435 440 445 Asp Lys Asp Arg Trp Gly ser ser Leu Gin Cys lie Lys Val Lys Asn 450 455 460 Asn Arg Leu Pro Tyr Val Ala Asp Lys Asp ser lie Ser Gin Glu lie 465 470 475 480 Phe Glu Asn Lys lie lie Thr Asp Glu Thr Asn val Gin Asn Tyr Ser 485 490 495 Asp Lys Phe ser Leu Asp Glu Ser lie Leu Asp Gly Gin Val pro lie 500 505 510 Asn Pro Glu lie val ASp Pro Leu Leu Pro Asn val Asn Met Glu Pro

    515 520 525

    Page 40

    2015268754 16 Dec 2015

    Leu Asn 530 Leu Pro P31003au-pct.txt Gly Glu Glu 535 lie Val Phe Tyr Asp 540 Asp He Thr Lys Tyr Val Asp Tyr Leu Asn ser Tyr Tyr Tyr Leu Glu Ser Gin Lys Leu 545 550 555 560 Ser Asn Asn val Glu Asn ile Thr Leu Thr Thr ser val Glu Gl u Ala 565 570 575 Leu Gly Tyr Ser Asn Lys lie Tyr Thr Phe Leu Pro Ser Leu Ala Glu 580 585 590 Lys Val Asn Lys Gly val Gin Ala Gly Leu Phe Leu Asn Trp Ala Asn 595 600 605 Glu Val val Glu Asp Phe Thr Thr Asn lie Met Lys Lys Asp Thr Leu 610 615 620 Asp Lys lie Ser Asp val Ser Val Ile lie Pro Tyr He Gly Pro Ala 625 630 635 640 Leu Asn lie Gly Asn Ser Ala Leu Arg Gly Asn Phe Asn Gin Ala Phe 645 650 655 Ala Thr Ala Gly Val Ala Phe Leu Leu GlU Gly Phe Pro Glu Phe Thr 660 665 670 ile pro Ala Leu Gly val Phe Thr Phe Tyr Ser Ser lie Gin Glu Arg 675 680 685 Glu Lys ile ile Lys Thr Ile Glu Asn Cys Leu Glu Gin Arg val Lys 690 695 700 Arg Trp Lys Asp Ser Tyr Gin Trp Met val Ser Asn Trp Leu Ser Arg 705 710 715 720 Ile Thr Thr Gin Phe Asn His Ile Asn Tyr Gin Met Tyr Asp Ser Leu 725 730 735 Ser Tyr Gin Ala Asp Ala Ile Lys Ala Lys Ile Asp Leu Glu Tyr Lys 740 745 750 Lys Tyr Ser Gly Ser Asp Lys Glu Asn lie Lys Ser Gin val Glu Asn 755 760 765 Leu Lys Asn Ser Leu Asp val Lys lie Ser Glu Ala Met Asn Asn Ile 770 775 780 Asn Lys Phe Ile Arg Glu Cys Ser Val Thr Tyr Leu Phe Lys Asn Met 785 790 795 800

    Page 41

    2015268754 16 Dec 2015

    P31003au~pct. txt

    Leu Pro Lys Val lie 805 Asp Glu Leu Asn Lys Phe Asp Leu Arg Thr Lys 810 815 Thr Glu Leu Ile Asn Leu lie Asp Ser His Asn lie Ile Leu val Gly 820 825 830 Glu Val Asp Arg Leu Lys Ala Lys val Asn Glu ser Phe Glu Asn Thr 835 840 845 Met Pro Phe Asn lie Phe Ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp 850 855 860 Ile lie Asn Glu Tyr Phe Asn Leu Glu Gly Gly Gly Gly ser Gly Gly 865 870 875 880 Gl y Gl y Ser Gly Gly Gly Gly ser Ala Leu Val Asn Ser Asp Ser Glu 885 890 895 Cys Pro Leu Ser His Asp Gly Tyr cys Leu His Asp Gly Val Cys Met 900 905 910 Tyr lie Glu Ala Leu Asp Lys Tyr Ala cys Asn cys val val Gly Tyr 915 920 925 lie Gly Glu Arg Cys Gln Tyr Arg Asp Leu Lys Trp Trp Glu Leu Arg 930 935 940 <210> 19 <211> 919 <212> PRT <213> Artificial f Sequence <220> <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 19 Thr Trp Pro val Lys Asp Phe Asn Tyr Ser Asp Pro val Asn Asp Asn 1 5 10 15 Asp Ile Leu Tyr Leu Arg Ile Pro Gln Asn Lys Leu lie Thr Thr Pro 20 25 30 val Lys Ala Phe Met Ile Thr Gln Asn ile Trp val lie Pro Glu Arg 35 40 45 Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 Gln Lys Asp Thr Phe Leu Lys Gly Ile Ile Lys Leu Phe Lys Arg lie Page 42

    2015268754 16 Dec 2015

    85 P31OO3au-pct.txt 90 95 Asn Glu Arg Asp Ile Gly Lys Lys Leu lie Asn Tyr Leu val Val Gly 100 105 110 ser Pro Phe Met Gly Asp Ser Ser Thr pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg Hi s Thr Thr Asn lie Ala val Glu Lys Phe Glu Asn Gly Ser 130 135 140 Trp Lys val Thr Asn lie lie Thr Pro ser Val Leu Ile Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly Gln 165 170 175 Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser lie Leu Lys 180 185 190 Val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp val Thr Ser Asn Gln 195 200 205 ser Ser Ala Val Leu Gly Lys Ser Ile Phe cys Met Asp Pro val Ile 210 215 220 Ala Leu Met His Glu Leu Thr Hi s Ser Leu His Gln Leu Tyr Gly lie 225 230 235 240 Asn Ile Pro Ser Asp Lys Arg lie Arg Pro Gln Val Ser Glu Gly Phe 245 250 255 Phe ser Gln Asp Gly Pro Asn Val Gln Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu Asp val Glu Ile Ile Pro Gln Ile Glu Arg ser Gln Leu 275 280 285 Arg Glu Lys Ala Leu Gly His Tyr Lys Asp lie Ala Lys Arg Leu Asn 290 295 300 Asn Ile Asn Lys Thr lie Pro ser Ser Trp Ile Ser Asn He Asp Lys 305 310 315 320 Tyr Lys Lys lie Phe ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Gly Asn Phe val val Asn Ile Asp Lys Phe Asn Ser Leu Tyr ser Asp 340 345 350 Leu Thr . Asn Val Met Ser Glu val val Tyr Ser Ser Gln Tyr Asn val Page 43

    P31OO3au-pct.txt

    355 360 365

    Ο

    Lys Asn Arg Thr His Tyr Phe Ser 375 Arg His Tyr Leu Pro val Phe Asn Ala Leu 400 O <D A _Asn kO 385 t—H 370 Asp Thr 380 Asp Asn 390 Ile Tyr Ile Arg 395 Asp Gly Phe ile Leu Thr xl- Asn Lys Gly Phe 405 Asn lie Glu Asn Ser 410 Gly Gin Asn lie Glu 415 Arg OO ^Asn Pro Ala Leu 420 Gin Lys Leu ser Ser 425 Glu Ser val val Asp 430 Leu Phe CH 'Ti -r-L. Thr O Lys Val 435 Cys val Asp Lys ser 440 Glu Glu Lys Leu Tyr 445 Asp Asp Asp ΓΗ Asp Lys 450 Asp Arg Trp Gly Ser 455 ser Leu Gin cys Ile 460 Lys Val Lys Asn Asn 465 Arg Leu Pro Tyr Val 470 Ala Asp Lys Asp Ser 475 lie Ser Gin Glu lie 480 phe Glu Asn Lys ile 485 lie Thr Asp Glu Thr 490 Asn val Gin Asn Tyr 495 Ser Asp Lys Phe Ser 500 Leu Asp Glu ser Ile 505 Leu Asp Gly Gin Val 510 Pro Ile Asn Pro Glu 515 Ile Val Asp Pro Leu 520 Leu Pro Asn Val Asn 525 Met Glu Pro Leu Asn 530 Leu Pro Gly Glu Glu 535 Ile val Phe Tyr Asp 540 Asp lie Thr Lys Tyr 545 Val Asp Tyr Leu Asn 550 Ser Tyr Tyr Tyr Leu 555 Glu Ser Gin Lys Leu 560 Ser Asn Asn val Glu 565 Asn lie Thr Leu Thr 570 Thr Ser Val Glu Glu 575 Ala Leu Gly Tyr ser 580 Asn Lys lie Tyr Thr 585 Phe Leu Pro Ser Leu 590 Ala Glu Lys Val Asn 595 Lys Gly Val Gin Ala 600 Gly Leu Phe Leu Asn 605 Trp Ala Asn Glu val 610 Val Glu Asp Phe Thr 615 Thr Asn lie Met Lys 620 Lys Asp Thr Leu Asp Lys Ile Ser Asp Val Ser Val Ile lie Pro Tyr Ile Gly Pro Ala

    Page 44

    2015268754 16 Dec 2015

    P31OO3au-pct.txt

    625 630 635 640

    Leu Asn lie Gly Asn Ser Ala Leu Arg Gly Asn Phe Asn Gin Ala Phe 645 650 655 Ala Thr Ala Gly Val Ala Phe Leu Leu Glu Gly Phe Pro Glu Phe Thr 660 665 670 lie Pro Ala Leu Gly val Phe Thr Phe Tyr Ser Ser lie Gin Glu Arg 675 680 685 Glu Lys lie lie Lys Thr lie Glu Asn cys Leu Glu Gin Arg val Lys 690 695 700 Arg Trp Lys Asp Ser Tyr Gin Trp Met val Ser Asn Trp Leu Ser Arg 705 710 715 720 He Thr Thr Gin Phe Asn Hi s lie Asn Tyr Gin Met Tyr Asp Ser Leu 725 730 735 Ser Tyr Gin Ala Asp Ala lie Lys Ala Lys lie Asp Leu Glu Tyr Lys 740 745 750 Lys Tyr Ser Gly ser Asp Lys Glu Asn lie Lys Ser Gin val Glu Asn 755 760 765 Leu Lys Asn ser Leu Asp val Lys lie Ser Glu Ala Met Asn Asn lie 770 775 780 Asn Lys Phe lie Arg Glu Cys ser Val Thr Tyr Leu Phe Lys Asn Met 785 790 795 800 Leu Pro Lys Val lie Asp Glu Leu Asn Lys Phe Asp Leu Arg Thr Lys 805 810 815 Thr Glu Leu lie Asn Leu lie Asp Ser Hi s Asn lie lie Leu Val Gly 820 825 830 Glu Val Asp Arg Leu Lys Ala Lys val Asn Glu Ser Phe Glu Asn Thr 835 840 845 Met Pro Phe Asn lie Phe Ser Tyr Thr Asn Asn ser Leu Leu Lys Asp 850 855 860 lie lie Asn Glu Tyr Phe Asn Leu Glu Gly Gly Gly Gly Ser Gly Gly 865 870 875 880 Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Val His Ser Asp Ala val 885 890 895 Phe Thr Asp Asn Tyr Thr Arg Leu Arg Lys Gin Met Ala Val Lys Lys

    Page 45

    5268754 16 Dec 2015

    900 P31OO3au-pct.txt 905 910 Tyr Leu Asn Ser Ile Leu Asn 915 <210> <211> <212> <213> 20 955 PRT Artificial sequence <220> <221> <223> source /note=Description of Artificial sequence: Synthetic

    polypeptide <400> 20

    Pro Ile Thr lie Asn Asn Phe Asn Tyr Ser Asp 10 Pro Val Asp Asn 15 Lys 1 5 Asn Ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu Pro 20 25 30 Glu Lys Ala Phe Arg lie Thr Gly Asn lie Trp val lie Pro Asp Arg 35 40 45 Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg val Thr 50 55 60 Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp Ser 65 70 75 80 Asp Lys Asp Thr Phe 85 Leu Lys Glu lie lie 90 Lys Leu Phe Lys Arg 95 Ile Asn ser Arg Glu lie Gly Glu Glu Leu ile Tyr Arg Leu Ser Thr Asp 100 105 110 lie Pro Phe Pro Gly Asn Asn Asn Thr Pro ile Asn Thr Phe Asp Phe 115 120 125 Asp val Asp Phe Asn ser Val Asp Val Lys Thr Arg Gin Gly Asn Asn 130 135 140 Trp Val Lys Thr Gly Ser Ile Asn Pro Ser val Ile Ile Thr Gly Pro 145 150 155 160 Arg Glu Asn lie lie Asp Pro Gl u Thr Ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Ala Ala Gin Glu Gly Phe Gly Ala Leu Ser lie Ile Ser 180 185 190 Ile Ser Pro Arg Phe Met Leu Thr Tyr ser Asn Ala Thr Asn Asp val

    195 200 205

    Page 46

    2015268754 16 Dec 2015

    P31OO3au-pct.txt

    Gly Glu Gly Arg phe ser Lys ser Glu Phe cys Met Asp Pro lie Leu 210 215 220 ile Leu Met Hi s Glu Leu Asn Hi s Ala Met His Asn Leu Tyr Gly lie 225 230 235 240 Ala lie Pro Asn Asp Gin Thr lie Ser Ser val Thr Ser Asn lie Phe 245 250 255 Tyr Ser Gin Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala Phe 260 265 270 Gly Gly Pro Thr Ile Asp Leu Ile Pro Lys Ser Ala Arg Lys Tyr Phe 275 280 285 Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser lie Ala Lys Arg Leu Asn 290 295 300 Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr lie Gly Glu 305 310 315 320 Tyr Lys Gin Lys Leu Ile Arg Lys Tyr Arg Phe val val Glu Ser Ser 325 330 335 Gly Glu val Thr val Asn Arg Asn Lys Phe val Glu Leu Tyr Asn Glu 340 345 350 Leu Thr Gin lie Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn Val 355 360 365 Gin Asn Arg Lys Ile Tyr Leu ser Asn Val Tyr Thr Pro val Thr Ala 370 375 380 Asn ile Leu Asp ASP Asn Val Tyr Asp lie Gin Asn Gly Phe Asn He 385 390 395 400 Pro Lys Ser Asn Leu Asn val Leu Phe Met Gly Gin Asn Leu Ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe Cys val Asp Ala lie Asp Gly Arg Ser Leu Tyr Asn Lys 435 440 445 Thr Leu Gin Cys Arg Glu Leu Leu Val Lys Asn Thr ASp Leu Pro Phe 450 455 460 lie Gly Asp ile ser Asp val Lys Thr ASp lie Phe Leu Arg Lys Asp 465 470 475 480

    Page 47

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    Ο (Ν

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    Ο

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    (Ν

    P31003au-pct.txt

    lie Asn Glu Glu Thr Glu Val lie Tyr Tyr Pro Asp 490 Asn val Ser 495 val 485 Asp Gin val Ile Leu Ser Lys Asn Thr Ser Glu Hl s Gly Gin Leu Asp 500 505 510 Leu Leu Tyr Pro ser lie Asp ser Glu Ser Glu lie Leu Pro Gly Glu 515 520 525 Asn Gin Val Phe Tyr ASp Asn Arg Thr Gin Asn Val Asp Tyr Leu Asn 530 535 540 Ser Tyr Tyr Tyr Leu Glu Ser Gin Lys Leu ser Asp Asn Val Glu Asp 545 550 555 560 Phe Thr Phe Thr Arg ser ile Glu Glu Ala Leu ASp Asn Ser Ala Lys 565 570 575 val Tyr Thr Tyr phe Pro Thr Leu Ala Asn Lys Val Asn Al a Gly val 580 585 590 Gin Gly Gly Leu phe Leu Met Trp Ala Asn Asp Val Val Glu Asp Phe 595 600 605 Thr Thr Asn lie Leu Arg Lys Asp Thr Leu Asp Lys ile Ser Asp Val 610 615 620 Ser Ala lie lie Pro Tyr Ile Gly Pro Ala Leu Asn Ile Ser Asn Ser 625 630 635 640 Val Arg Arg Gly Asn Phe Thr Glu Ala Phe Ala Val Thr Gly val Thr 645 650 655 lie Leu Leu Glu Ala Phe Pro Glu Phe Thr lie Pro Ala Leu Gly Ala 660 665 670 Phe Val lie Tyr Ser Lys Val Gin Glu Arg Asn Glu lie Ile Lys Thr 675 680 685 lie Asp Asn Cys Leu Glu Gin Arg Ile Lys Arg Trp Lys Asp Ser Tyr 690 695 700 Glu Trp Met Met Gly Thr Trp Leu Ser Arg lie Ile Thr Gin Phe Asn 705 710 715 720 Asn lie ser Tyr Gin Met Tyr Asp Ser Leu Asn Tyr Gin Ala Gly Ala 725 730 735 lie Lys Ala Lys ile Asp Leu Glu Tyr Lys Lys Tyr Ser Gly ser Asp 740 745 750

    Page 48

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    P31OO3au~pct.txt

    Lys Glu Asn Ile Lys Ser Gin val Glu Asn Leu Lys Asn ser Leu Asp 755 760 765 Val Lys lie Ser Glu Ala Met Asn Asn lie Asn Lys Phe Ile Arg Glu 770 775 780 Cys Ser val Thr Tyr Leu Phe Lys Asn Met Leu pro Lys val lie Asp 785 790 795 800 Glu Leu Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu Ile Asn Leu 805 810 815 Ile Asp Ser Hi s Asn Ile Ile Leu Val Gly Glu Val Asp Lys Leu Lys 820 825 830 Ala Lys Val Asn Asn ser Phe Gin Asn Thr Ile Pro Phe Asn lie Phe 835 840 845 Ser Tyr Thr Asn Asn ser Leu Leu Lys Asp Ile lie Asn Glu Tyr Phe 850 855 860 Asn Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 865 870 875 880 Gly ser Ala Leu val Gly Pro Glu Thr Leu cys Gly Al a Glu Leu Val 885 890 895 Asp Ala Leu Gin Phe val Cys Gly Asp Arg Gly Phe Tyr Phe Asn Lys 900 905 910 Pro Thr Gly Tyr Gly ser Ser Ser Arg Arg Ala Pro Gin Thr Gly lie 915 920 925 val Asp Glu cys Cys Phe Arg Ser Cys Asp Leu Arg Arg Leu Glu Met 930 935 940 Tyr cys Ala pro Leu Lys Pro Ala Lys Ser Ala

    945 950 955 <210> 21 <211> 961 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polypeptide <400> 21

    Thr Trp Pro val Lys Asp Phe Asn Tyr Ser Asp Pro Val Asn Asp Asn 15 10 15

    Page 49

    P31003au~pct.txt

    IT)

    ΞΓ Asp lie Leu Tyr Leu Arg lie Pro Gln 25 Asn Lys Leu Ile Thr 30 Thr pro CN 20 o Ile <D val Lys Ala Phe Met lie Thr Gln Asn lie Trp Val Pro Glu Arg Q 35 40 45 Phe ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro pro Arg Pro Thr 50 55 60 Xt tn o Ser Lys Tyr Gln ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 00 CN IT) Gln Lys Asp Thr Phe Leu Lys Gly Ile lie Lys Leu Phe Lys Arg lie 85 90 95 1—1 o CN Asn Glu Arg Asp lie Gly Lys Lys Leu ile Asn Tyr Leu Val Val Gly 100 105 110 Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg Hi s Thr Thr Asn lie Ala val Glu Lys Phe Glu Asn Gly ser 130 135 140 Trp Lys val Thr Asn lie lie Thr pro ser Val Leu Ile Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu Asp Tyr Thr Ala ser Leu Thr Leu Gln Gly Gln 165 170 175 Gln Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu ser lie Leu Lys 180 185 190 val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp val Thr Ser Asn Gln 195 200 205 Ser Ser Ala Val Leu Gly Lys ser ile Phe cys Met Asp Pro val ile 210 215 220 Ala Leu Met Hi s Glu Leu Thr Hi s ser Leu His Gln Leu Tyr Gly lie 225 230 235 240 Asn lie Pro Ser Asp Lys Arg lie Arg Pro Gln Val Ser Glu Gly Phe 245 250 255 Phe ser Gln Asp Gly Pro Asn val Gln Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu Asp val Glu Ile Ile Pro Gln lie Glu Arg Ser Gln Leu 275 280 285

    Page 50

    2015268754 16 Dec 2015

    Arg Glu 290 Lys Ala Leu Gly Hi s 295 Tyr Asn 305 lie Asn Lys Thr lie 310 Pro Ser Tyr Lys Lys lie Phe 325 Ser Glu Lys Gly Asn Phe val 340 Val Asn lie Asp Leu Thr Asn 355 val Met Ser Glu Val 360 Lys Asn 370 Arg Thr Hi s Tyr Phe 375 Ser Asn 385 lie Leu Asp Asp Asn 390 lie Tyr Thr Asn Lys Gly Phe 405 Asn lie Glu Asn Pro Ala Leu 420 Gin Lys Leu ser Thr Lys Val 435 cys val Asp Lys Ser 440 Asp Lys 450 Asp Arg Trp Gly Ser 455 Ser Asn 465 Arg Leu Pro Tyr val 470 Ala Asp Phe Glu Asn Lys lie 485 lie Thr Asp Asp Lys Phe Ser 500 Leu Asp Glu Ser Asn Pro Glu 515 lie Val Asp Pro Leu 520 Leu Asn 530 Leu Pro Gly Glu Glu 535 lie Tyr 545 Val Asp Tyr Leu Asn 550 Ser Tyr

    P31003au-pct.txt

    Lys Asp lie Ala 300 Lys Arg Leu Asn Ser Trp He 315 ser Asn lie Asp Lys 320 Tyr Asn 330 Phe Asp Lys Asp Asn 335 Thr Lys 345 Phe Asn Ser Leu Tyr 350 Ser Asp Val Tyr Ser Ser Gin 365 Tyr Asn Val Arg Hi s Tyr Leu 380 Pro val Phe Ala Thr lie Arg 395 ASp Gly Phe Asn Leu 400 Asn Ser 410 Gly Gin Asn lie Glu 415 Arg Ser 425 Glu ser val val Asp 430 Leu Phe Glu Glu Lys Leu Tyr 445 Asp Asp Asp Leu Gin Cys lie 460 Lys Val Lys Asn Lys Asp Ser 475 lie Ser Gin Glu lie 480 Glu Thr 490 Asn val Gin Asn Tyr 495 Ser lie 505 Leu Asp Gly Gin Val 510 pro He Leu Pro Asn Val Asn 525 Met Glu Pro val Phe Tyr Asp 540 Asp lie Thr Lys Tyr Tyr Leu 555 Glu Ser Gin Lys Leu 560

    Page 51

    P31OO3au-pct.txt on ο

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    Ser Asn Asn val Glu Asn lie Thr Leu Thr Thr Ser val Glu Glu 575 Ala 565 570 Leu Gly Tyr ser Asn Lys Ile Tyr Thr Phe Leu Pro Ser Leu Ala Glu 580 585 590 Lys Val Asn Lys Gly val Gln Ala Gly Leu Phe Leu Asn Trp Ala Asn 595 600 605 Glu val Val Glu Asp Phe Thr Thr Asn lie Met Lys Lys Asp Thr Leu 610 615 620 Asp Lys Ile ser Asp val Ser val lie lie Pro Tyr Ile Gly pro Ala 625 630 635 640 Leu Asn lie Gly Asn Ser Ala Leu Arg Gly Asn Phe Asn Gln Ala Phe 645 650 655 Ala Thr Ala Gly Val Ala Phe Leu Leu Glu Gly Phe Pro Glu Phe Thr 660 665 670 ile Pro Ala Leu Gly Val Phe Thr Phe Tyr Ser Ser Ile Gln Glu Arg 675 680 685 Glu Lys lie lie Lys Thr He Glu Asn cys Leu Glu Gln Arg val Lys 690 695 700 Arg Trp Lys Asp Ser Tyr Gln Trp Met val Ser Asn Trp Leu Ser Arg 705 710 715 720 ile Thr Thr Gln Phe Asn Hi s lie Asn Tyr Gln Met Tyr Asp Ser Leu 725 730 735 Ser Tyr Gln Ala Asp Ala Ile Lys Ala Lys ile Asp Leu Glu Tyr Lys 740 745 750 Lys Tyr ser Gly Ser Asp Lys Glu Asn ile Lys Ser Gln Val Glu Asn 755 760 765 Leu Lys Asn Ser Leu Asp val Lys lie Ser Glu Ala Met Asn Asn Ile 770 775 780 Asn Lys Phe lie Arg Glu cys ser val Thr Tyr Leu Phe Lys Asn Met 785 790 795 800 Leu Pro Lys val lie Asp Glu Leu Asn Lys Phe Asp Leu Arg Thr Lys 805 810 815 Thr Glu Leu lie Asn Leu lie Asp Ser Hi s Asn Ile lie Leu Val Gly 820 825 830

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    P31OO3au-pct.txt

    Glu val Asp Arg Leu 835 Lys Ala Lys 840 Val Asn Glu Ser Phe 845 Glu Asn Thr Met Pro Phe Asn lie Phe ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp 850 855 860 Ile lie Asn Glu Tyr Phe Asn Leu Glu Gly Gly Gly Gly Ser Gly Gly 865 870 875 880 Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Val Gly Pro Glu Thr Leu 885 890 895 Cys Gly Ala Glu Leu val Asp Ala Leu Gin Phe Val Cys Gly Asp Arg 900 905 910 Gly Phe Tyr Phe Asn Lys Pro Thr Gly Tyr Gly Ser ser Ser Arg Arg 915 920 925 Ala Pro Gin Thr Gly ile Val Asp Glu Cys Cys Phe Arg Ser Cys Asp 930 935 940 Leu Arg Arg Leu Glu Met Tyr Cys Ala Pro Leu Lys Pro Ala Lys Ser 945 950 955 960 Ala <210> 22 <211> 913 <212> PRT <213> Artificial Sequence <220> <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 22 pro ile Thr lie Asn Asn Phe Asn Tyr Ser Asp Pro val Asp Asn Lys 1 5 10 15 Asn lie Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu Pro 20 25 30 Glu Lys Ala Phe Arg lie Thr Gly Asn lie Trp val lie Pro Asp Arg 35 40 45 Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro pro Arg val Thr 50 55 60 Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp Ser 65 70 75 80

    Page 53

    IT)

    Ο

    CN

    Ο <υ

    Q vt oo kO

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    CN

    ASP Lys Asp Thr phe Leu Lys Glu 85 P31003au-pct. txt lie lie 90 Lys Leu Phe Lys Arg 95 Ile Asn ser Arg Glu ile Gly Glu Glu Leu ile Tyr Arg Leu Ser Thr Asp 100 105 110 lie Pro Phe Pro Gly Asn Asn Asn Thr Pro lie Asn Thr Phe Asp Phe 115 120 125 Asp val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gln Gly Asn Asn 130 135 140 Trp val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly pro 145 150 155 160 Arg Glu Asn lie lie Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Ala Ala Gln Glu Gly phe Gly Ala Leu ser lie Ile Ser 180 185 190 lie Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp val 195 200 205 Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile Leu 210 215 220 Ile Leu Met His Glu Leu Asn His Ala Met His Asn Leu Tyr Gly ile 225 230 235 240 Ala lie Pro Asn Asp Gln Thr lie ser Ser Val Thr Ser Asn ile Phe 245 250 255 Tyr ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu lie Tyr Ala Phe 260 265 270 Gly Gly Pro Thr Ile Asp Leu lie Pro Lys ser Ala Arg Lys Tyr Phe 275 280 285 Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu Asn 290 295 300 Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly Glu 305 310 315 320 Tyr Lys Gln Lys Leu lie Arg Lys Tyr Arg Phe val val Glu ser Ser 325 330 335 Gly Glu val Thr Val Asn Arg Asn Lys Phe val Glu Leu Tyr Asn Glu 340 345 350

    Page 54

    2015268754 16 Dec 2015

    P31 003a u-pc t.tx t Leu Thr Gln 355 lie Phe Thr Glu Phe 360 Asn Tyr Ala Lys lie 365 Tyr Asn Val Gln Asn Arg Lys lie Tyr Leu Ser Asn Val Tyr Thr Pro Val Thr Ala 370 375 380 Asn lie Leu Asp Asp Asn val Tyr Asp lie Gln Asn Gly Phe Asn lie 385 390 395 400 pro Lys ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe Cys val Asp Ala Ile ASP Gly Arg ser Leu Tyr Asn Lys 435 440 445 Thr Leu Gln cys Arg Glu Leu Leu val Lys Asn Thr Asp Leu Pro Phe 450 455 460 lie Gly Asp lie Ser Asp Val Lys Thr Asp Ile Phe Leu Arg Lys Asp 465 470 475 480 Ile Asn Glu Glu Thr Glu Val Ile Tyr Tyr Pro Asp Asn Val Ser val 485 490 495 Asp Gln val lie Leu ser Lys Asn Thr Ser Glu Hi s Gly Gln Leu Asp 500 505 510 Leu Leu Tyr Pro Ser Ile Asp Ser Glu ser Glu lie Leu Pro Gly Glu 515 520 525 Asn Gln val Phe Tyr Asp Asn Arg Thr Gln Asn val Asp Tyr Leu Asn 530 535 540 ser Tyr Tyr Tyr Leu Glu Ser Gln Lys Leu Ser Asp Asn val Glu Asp 545 550 555 560 Phe Thr Phe Thr Arg Ser lie Glu Glu Ala Leu Asp Asn ser Ala Lys 565 570 575 val Tyr Thr Tyr Phe pro Thr Leu Ala Asn Lys val Asn Ala Gly val 580 585 590 Gln Gly Gly Leu Phe Leu Met Trp Ala Asn Asp val Val Glu Asp Phe 595 600 605 Thr Thr Asn Ile Leu Arg Lys Asp Thr Leu Asp Lys Ile Ser Asp Val 610 615 620

    Page 55

    P3K 303ai j-pct.txt *4 i ser Ala lie Ile Pro Tyr lie Gly Pro Ala Leu Asn lie Ser Asn Ser o 625 630 635 640 <N o Val Arg Arg Gly Asn phe Thr Glu Al a Phe Ala Val Thr Gly val Thr <D Q 645 650 655 lie Leu Leu Glu Ala phe Pro Glu Phe Thr ile Pro Ala Leu Gly Ala 660 665 670 T-f- Phe val lie Tyr ser Lys val Gin Glu Arg Asn Glu lie Ile Lys Thr xl •X'j 675 680 685 l> oo kO ile Asp Asn cys Leu Glu Gin Arg lie Lys Arg Trp Lys Asp Ser Tyr Cfl 690 695 700 LA r-H r-> Glu Trp Met Met Gly Thr Trp Leu Ser Arg ile lie Thr Gin Phe Asn CA 705 710 715 720 Asn Ile ser Tyr Gin Met Tyr Asp Ser Leu Asn Tyr Gin Ala Gly Ala 725 730 735 lie Lys Ala Lys Ile Asp Leu Glu Tyr Lys Lys Tyr Ser Gly ser Asp 740 745 750 Lys Glu Asn Ile Lys ser Gin Val Glu Asn Leu Lys Asn Ser Leu Asp 755 760 765 Val Lys Ile Ser Glu Ala Met Asn Asn Ile Asn Lys Phe Ile Arg Glu 770 775 780 Cys ser Val Thr Tyr Leu Phe Lys Asn Met Leu Pro Lys Val lie Asp 785 790 795 800 Glu Leu Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu Ile Asn Leu 805 810 815 Ile Asp Ser Hi s Asn Ile Ile Leu Val Gly Glu val Asp Lys Leu Lys 820 825 830 Ala Lys val Asn Asn Ser Phe Gin Asn Thr lie Pro Phe Asn lie Phe 835 840 845 ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp lie He Asn Glu Tyr Phe 850 855 860 Asn Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 865 870 875 880 Gly Ser Ala Leu val His Ser Asp Ala Val Phe Thr Asp Asn Tyr Thr 885 890 895

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    P31OO3au-pct.txt

    Arg Leu Arg Lys Gln Met Ala Val Lys Lys Tyr Leu Asn Ser lie Leu 900 905 910

    Asn <210> 23 <211> 977 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide

    <400> 23 Phe Asn Tyr ser 10 Asp Pro val Asp Asn 15 Lys pro 1 lie Thr lie Asn Asn 5 Asn Ile Leu Tyr Leu Asp Thr Hi s Leu Asn Thr Leu Ala Asn Glu Pro 20 25 30 Glu Lys Ala Phe Arg Ile Thr Gly Asn lie Trp Val lie Pro Asp Arg 35 40 45 Phe Ser Arg Asn ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val Thr 50 55 60 Ser Pro Lys ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp ser 65 70 75 80 Asp Lys Asp Thr Phe Leu Lys Glu lie lie Lys Leu Phe Lys Arg Ile 85 90 95 Asn Ser Arg Glu Ile Gly Glu Glu Leu lie Tyr Arg Leu Ser Thr Asp 100 105 110 lie Pro Phe Pro Gly Asn Asn Asn Thr Pro Ile Asn Thr Phe Asp Phe 115 120 125 Asp Val Asp Phe Asn Ser Val ASP Val Lys Thr Arg Gln Gly Asn Asn 130 135 140 Trp Val Lys Thr Gly ser ile Asn Pro ser val Ile Ile Thr Gly Pro 145 150 155 160 Arg Glu Asn He Ile Asp Pro Glu Thr ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser lie lie Ser 180 185 190 lie Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp val

    Page 57

    P31003au-pct.txt

    195 200 205

    Ο

    (Μ Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile Leu Ο 210 215 220 <υ ο Ω lie Leu Met His Glu Leu Asn Hi s Ala Met His Asn Leu Tyr Gly Ile 90 225 230 235 240 Η Ala lie pro Asn Asp Gln Thr lie Ser Ser Val Thr Ser Asn lie Phe 245 250 255 ιη ο οο 90 Tyr Ser Gln Tyr Asn val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala Phe 260 265 270 (Μ Gly Gly Pro Thr Ile Asp Leu lie Pro Lys Ser Ala Arg Lys Tyr Phe ο 275 280 285 (Μ Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser Ile Ala Lys Arg Leu Asn 290 295 300 Ser Ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly Glu 305 310 315 320 Tyr Lys Gln Lys Leu 325 Ile Arg Lys Tyr Arg 330 Phe val val Glu ser 335 Ser Gly Glu val Thr Val Asn Arg Asn Lys Phe val Glu Leu Tyr Asn Glu 340 345 350 Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys lie Tyr Asn val 355 360 365 Gin Asn Arg Lys lie Tyr Leu Ser Asn val Tyr Thr Pro val Thr Ala 370 375 380 Asn Ile Leu Asp Asp Asn Val Tyr Asp Ile Gln Asn Gly Phe Asn lie 385 390 395 400 Pro Lys Ser Asn Leu Asn val Leu Phe Met Gly Gln Asn Leu Ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe Cys Val Asp Ala ile Asp Gly Arg Ser Leu Tyr Asn Lys 435 440 445 Thr Leu Gln cys Arg Glu Leu Leu Val Lys Asn Thr Asp Leu Pro Phe 450 455 460 lie Gly Asp lie Ser Asp Val Lys Thr Asp Ile Phe Leu Arg Lys Asp

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    P31003au-pct.txt

    465 470 475 480

    lie Asn Glu Glu Thr Glu Val 485 lie Tyr Tyr 490 Pro Asp Asn val Ser 495 val ASP Gin Val lie Leu Ser Lys Asn Thr Ser Glu His Gly Gin Leu Asp 500 505 510 Leu Leu Tyr Pro Ser lie Asp Ser Glu Ser Glu lie Leu Pro Gly Glu 515 520 525 Asn Gin Val Phe Tyr Asp Asn Arg Thr Gin Asn Val ASp Tyr Leu Asn 530 535 540 Ser Tyr Tyr Tyr Leu Glu ser Gin Lys Leu Ser Asp Asn val Glu Asp 545 550 555 560 Phe Thr Phe Thr Arg Ser lie Glu Glu Ala Leu ASp Asn ser Ala Lys 565 570 575 Val Tyr Thr Tyr Phe pro Thr Leu Ala Asn Lys Val Asn Ala Gly Val 580 585 590 Gin Gly Gly Leu Phe Leu Met Trp Ala Asn Asp Val val Glu Asp Phe 595 600 605 Thr Thr Asn lie Leu Arg Lys Asp Thr Leu Asp Lys lie Ser Asp val 610 615 620 ser Ala lie lie Pro Tyr lie Gly Pro Ala Leu Asn lie Ser Asn Ser 625 630 635 640 val Arg Arg Gly Asn Phe Thr Glu Ala Phe Ala Val Thr Gly val Thr 645 650 655 lie Leu Leu Glu Ala Phe Pro Glu Phe Thr lie Pro Ala Leu Gly Ala 660 665 670 Phe val He Tyr ser Lys val Gin Glu Arg Asn Glu lie lie Lys Thr 675 680 685 lie Asp Asn Cys Leu Glu Gin Arg lie Lys Arg Trp Lys Asp ser Tyr 690 695 700 Glu Trp Met Met Gly Thr Trp Leu Ser Arg lie lie Thr Gin Phe Asn 705 710 715 720 Asn He Ser Tyr Gin Met Tyr Asp Ser Leu Asn Tyr Gin Ala Gly Ala 725 730 735 lie Lys Ala Lys lie Asp Leu Glu Tyr Lys Lys Tyr ser Gly ser Asp

    Page 59

    2015268754 16 Dec 2015

    P31OO3au-pct.txt

    740 745 750

    Lys Glu Asn lie Lys Ser Gin Val 760 Glu Asn Leu Lys Asn 765 Ser Leu ASP 755 val Lys lie Ser Glu Ala Met Asn Asn He Asn Lys Phe lie Arg Glu 770 775 780 cys ser Val Thr Tyr Leu Phe Lys Asn Met Leu Pro Lys Val He Asp 785 790 795 800 Glu Leu Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu lie Asn Leu 805 810 815 lie Asp Ser His Asn lie lie Leu Val Gly Glu Val Asp Lys Leu Lys 820 825 830 Ala Lys Val Asn Asn Ser Phe Gin Asn Thr lie Pro Phe Asn lie Phe 835 840 845 ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp lie lie Asn Glu Tyr Phe 850 855 860 Asn Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 865 870 875 880 Gly Ser Ala Leu Val Met Lys Pro lie Gin Lys Leu Leu Ala Gly Leu 885 890 895 lie Leu Leu Thr Trp cys Val Glu Gly Cys ser Ser Gin His Trp Ser 900 905 910 Tyr Gly Leu Arg Pro Gly Gly Lys Arg Asp Ala Glu Asn Leu lie Asp 915 920 925 Ser Phe Gin Glu lie val Lys Glu Val Gly Gin Leu Ala Glu Thr Gin 930 935 940 Arg Phe Glu Cys Thr Thr His Gin Pro Arg Ser Pro Leu Arg Asp Leu 945 950 955 960 Lys Gly Ala Leu Glu Ser Leu lie Glu Glu Glu Thr Gly Gin Lys Lys

    965 970 975 lie <210> 24 <211> 988 <212> PRT <213> Artificial sequence

    Page 60

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 24

    Thr 1 Trp Pro val Lys Asp 5 Phe Asn Tyr Ser Asp Pro 10 val Asn Asp 15 Asn Asp lie Leu Tyr Leu Arg lie Pro Gin Asn Lys Leu lie Thr Thr Pro 20 25 30 Val Lys Ala Phe Met lie Thr Gin Asn lie Trp Val lie Pro Glu Arg 35 40 45 Phe Ser Ser Asp Thr Asn pro Ser Leu Ser Lys Pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gin Ser Tyr Tyr Asp Pro Ser Tyr Leu ser Thr Asp Glu 65 70 75 80 Gin Lys Asp Thr Phe Leu Lys Gly lie lie Lys Leu Phe Lys Arg lie 85 90 95 Asn Glu Arg Asp lie Gly Lys Lys Leu lie Asn Tyr Leu val val Gly 100 105 110 Ser Pro Phe Met Gly Asp ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg His Thr Thr Asn lie Ala val Glu Lys Phe Glu Asn Gly Ser 130 135 140 Trp Lys val Thr Asn lie lie Thr Pro Ser Val Leu lie Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu ASp Tyr Thr Ala Ser Leu Thr Leu Gin Gly Gin 165 170 175 Gin Ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser lie Leu Lys 180 185 190 val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp val Thr Ser Asn Gin 195 200 205 ser Ser Ala val Leu Gly Lys Ser lie Phe cys Met Asp Pro val lie 210 215 220 Ala Leu Met Hi s Glu Leu Thr His Ser Leu Hi s Gin Leu Tyr Gly lie 225 230 235 240 Asn lie Pro Ser Asp Lys Arg lie Arg Pro Gin val Ser Glu Gly Phe 245 250 255

    Page 61

    P31OO3au-pct.txt in r-H o

    (N o

    CD

    Ω

    Xl^- in o

    oo kO

    CM in ©

    (N

    Phe ser Gin Asp Gly Pro Asn Val Gin Phe Glu Glu Leu 265 Tyr Thr 270 Phe 260 Gly Gly Leu Asp val Glu lie He pro Gin He Glu Arg Ser Gin Leu 275 280 285 Arg Glu Lys Ala Leu Gly His Tyr Lys Asp He Al a Lys Arg Leu Asn 290 295 300 Asn lie Asn Lys Thr lie Pro Ser Ser Trp He ser Asn He Asp Lys 305 310 315 320 Tyr Lys Lys lie Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Gly Asn Phe Val Val Asn He Asp Lys Phe Asn Ser Leu Tyr Ser Asp 340 345 350 Leu Thr Asn val Met Ser Glu val val Tyr ser Ser Gin Tyr Asn val 355 360 365 Lys Asn Arg Thr Hi s Tyr Phe Ser Arg Hi s Tyr Leu Pro Val Phe Ala 370 375 380 Asn lie Leu Asp Asp Asn lie Tyr Thr lie Arg Asp Gly Phe Asn Leu 385 390 395 400 Thr Asn Lys Gly Phe Asn He Glu Asn ser Gly Gin Asn He Glu Arg 405 410 415 Asn pro Ala Leu Gin Lys Leu Ser ser Glu ser val val Asp Leu Phe 420 425 430 Thr Lys val cys val ASp Lys Ser Glu Glu Lys Leu Tyr Asp Asp Asp 435 440 445 Asp Lys Asp Arg Trp Gly Ser Ser Leu Gin Cys lie Lys Val Lys Asn 450 455 460 Asn Arg Leu Pro Tyr val Ala Asp Lys Asp ser lie Ser Gin Glu lie 465 470 475 480 Phe Glu Asn Lys lie lie Thr Asp Glu Thr Asn val Gin Asn Tyr ser 485 490 495 Asp Lys Phe Ser Leu ASp Glu Ser lie Leu Asp Gly Gin Val Pro lie 500 505 510 Asn pro Glu lie val Asp Pro Leu Leu Pro Asn val Asn Met Glu Pro

    515 520 525

    Page 62

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Leu Asn Leu 530 Pro Gly Glu Glu Ile val 535 Phe Tyr Asp 540 Asp ile Thr Lys Tyr val Asp Tyr Leu Asn ser Tyr Tyr Tyr Leu Glu ser Gin Lys Leu 545 550 555 560 Ser Asn Asn val Glu Asn lie Thr Leu Thr Thr Ser val Glu Glu Ala 565 570 575 Leu Gly Tyr Ser Asn Lys Ile Tyr Thr Phe Leu Pro Ser Leu Ala Glu 580 585 590 Lys Val Asn Lys Gly Val Gin Ala Gly Leu Phe Leu Asn Trp Ala Asn 595 600 605 Glu Val val Glu Asp Phe Thr Thr Asn Ile Met Lys Lys Asp Thr Leu 610 615 620 Asp Lys lie Ser Asp val Ser val lie lie Pro Tyr lie Gly Pro Ala 625 630 635 640 Leu Asn Ile Gly Asn Ser Ala Leu Arg Gly Asn Phe Asn Gin Ala Phe 645 650 655 Al a Thr Ala Gly val Ala Phe Leu Leu Glu Gly Phe pro Glu Phe Thr 660 665 670 Ile Pro Ala Leu Gly val Phe Thr Phe Tyr Ser Ser lie Gin Glu Arg 675 680 685 Glu Lys lie lie Lys Thr ile Glu Asn cys Leu Glu Gin Arg Val Lys 690 695 700 Arg Trp Lys Asp Ser Tyr Gin Trp Met val Ser Asn Trp Leu ser Arg 705 710 715 720 Ile Thr Thr Gin Phe Asn His Ile Asn Tyr Gin Met Tyr Asp ser Leu 725 730 735 Ser Tyr Gin Ala Asp Ala lie Lys Ala Lys lie Asp Leu Glu Tyr Lys 740 745 750 Lys Tyr ser Gly Ser Asp Lys Glu Asn Ile Lys Ser Gin Val Glu Asn 755 760 765 Leu Lys Asn Ser Leu Asp Val Lys lie Ser Glu Ala Met Asn Asn lie 770 775 780 Asn Lys Phe lie Arg Glu Cys Ser val Thr Tyr Leu Phe Lys Asn Met 785 790 795 800

    Page 63

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Leu Pro Lys Val Ile Asp Glu Leu Asn Lys Phe Asp Leu Arg Thr Lys 805 810 815 Thr Glu Leu lie Asn Leu ile Asp Ser His Asn lie ile Leu Val Gly 820 825 830 Glu val Asp Arg Leu Lys Ala Lys Val Asn Glu Ser Phe Glu Asn Thr 835 840 845 Met Pro Phe Asn lie Phe Ser Tyr Thr Asn Asn ser Leu Leu Lys Asp 850 855 860 lie ile Asn Glu Tyr Phe Asn Leu Glu Gly Gly Gly Gly Ser Gly Gly 865 870 875 880 Gly Gly Ser Gly Gly 885 Gly Gly Ser Gly Gly 890 Gly Gly ser Ala Leu 895 Val Met Lys Pro Ile Gin Lys Leu Leu Ala Gly Leu lie Leu Leu Thr Trp 900 905 910 cys val Glu Gly cys Ser Ser Gin Hi s Trp Ser Tyr Gly Leu Arg Pro 915 920 925 Gly Gly Lys Arg Asp Ala Glu Asn Leu Ile Asp ser Phe Gin Glu He 930 935 940 val Lys Glu val Gly Gin Leu Ala Glu Thr Gin Arg Phe Glu cys Thr 945 950 955 960 Thr Hi s Gin Pro Arg Ser pro Leu Arg Asp Leu Lys Gly Ala Leu Glu 965 970 975 Ser Leu lie Glu Glu Glu Thr Gly Gin Lys Lys lie 980 985

    <210> 25 <211> 901 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 25

    Thr Trp Pro val Lys Asp Phe Asn Tyr Ser Asp Pro val Asn Asp Asn 1 5 10 15 Asp Ile Leu Tyr Leu Arg He Pro Gin Asn Lys Leu lie Thr Thr Pro

    20 25 30

    Page 64

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    val Lys Ala 35 Phe Met lie Thr Gin Asn lie Trp 40 val Ile 45 Pro Glu Arg Phe Ser ser Asp Thr Asn Pro Ser Leu ser Lys pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gin Ser Tyr Tyr Asp Pro ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 Gin Lys Asp Thr Phe Leu Lys Gly lie lie Lys Leu Phe Lys Arg He 85 90 95 Asn Glu Arg Asp Ile Gly Lys Lys Leu ile Asn Tyr Leu val Val Gly 100 105 110 Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg Hi s Thr Thr Asn ile Ala Val Glu Lys phe Glu Asn Gly Ser 130 135 140 Trp Lys val Thr Asn lie Ile Thr Pro ser Val Leu lie Phe Gly Pro 145 150 155 160 Leu pro Asn lie Leu Asp Tyr Thr Ala ser Leu Thr Leu Gin Gly Gin 165 170 175 Gin ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser lie Leu Lys 180 185 190 val Ala Pro Glu Phe Leu Leu Thr Phe ser Asp val Thr ser Asn Gin 195 200 205 Ser Ser Ala Val Leu Gly Lys Ser Ile Phe Cys Met ASp Pro val ile 210 215 220 Ala Leu Met His Glu Leu Thr His Ser Leu Hi s Gin Leu Tyr Gly Ile 225 230 235 240 Asn lie Pro Ser Asp Lys Arg lie Arg Pro Gin val ser Glu Gly Phe 245 250 255 Phe Ser Gin Asp Gly Pro Asn val Gin Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu Asp Val Glu lie lie Pro Gin Ile Glu Arg Ser Gin Leu 275 280 285 Arg Glu Lys Ala Leu Gly His Tyr Lys Asp Ile Ala Lys Arg Leu Asn 290 295 300

    Page 65

    P31003au-pct.txt ι/Ί o

    <N !\sn

    305

    Q ^1^- in oo

    CH in o

    <N

    Gly

    Leu

    Lys

    Asn

    385

    Thr

    Asn

    Thr

    Asp

    Asn

    465

    Phe

    Asp

    Asn

    Leu

    Tyr

    545 ser

    lie Asn Lys Thr lie Pro 310 Ser Ser Trp lie Ser Asn lie 315 Asp Lys 320 Lys Lys lie Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Asn Phe val Val Asn lie ASp Lys Phe Asn ser Leu Tyr Ser Asp 340 345 350 Thr Asn Val Met Ser Glu val Val Tyr Ser ser Gin Tyr Asn Val 355 360 365 Asn Arg Thr Hi s Tyr Phe Ser Arg Hi s Tyr Leu Pro val Phe Ala 370 375 380 lie Leu Asp Asp Asn lie Tyr Thr lie Arg Asp Gly Phe Asn Leu 390 395 400 Asn Lys Gly Phe Asn lie Glu Asn Ser Gly Gin Asn lie Glu Arg 405 410 415 pro Ala Leu Gin Lys Leu Ser Ser Glu Ser Val val Asp Leu Phe 420 425 430 Lys val cys Val Asp Lys Ser Glu Glu Lys Leu Tyr Asp Asp Asp 435 440 445 Lys Asp Arg Trp Gly Ser Ser Leu Gin cys lie Lys val Lys Asn 450 455 460 Arg Leu Pro Tyr Val 470 Ala Asp Lys Asp Ser 475 lie ser Gin Glu lie 480 Glu Asn Lys lie lie Thr Asp Glu Thr Asn val Gin Asn Tyr Ser 485 490 495 Lys Phe ser Leu Asp Glu Ser lie Leu Asp Gly Gin val pro lie 500 505 510 Pro Glu lie val Asp Pro Leu Leu Pro Asn val Asn Met Glu Pro 515 520 525 Asn Leu Pro Gly Glu Glu lie Val Phe Tyr Asp Asp lie Thr Lys 530 535 540 Val Asp Tyr Leu Asn Ser Tyr Tyr Tyr Leu Glu ser Gin Lys Leu 550 555 560 Asn Asn Val Glu Asn lie Thr Leu Thr Thr Ser Val Glu Glu Ala

    565 570 575

    Page 66

    2015268754 16 Dec 2015

    P31003au~pct.txt

    Leu Gly Tyr Ser Asn Lys lie Tyr Thr Phe Leu Pro Ser Leu Ala Glu 580 585 590 Lys val Asn Lys Gly val Gin Ala Gly Leu phe Leu Asn Trp Ala Asn 595 600 605 Glu val Val Glu Asp Phe Thr Thr Asn lie Met Lys Lys Asp Thr Leu 610 615 620 Asp Lys Ile ser Asp Val Ser val lie He Pro Tyr Ile Gly pro Ala 625 630 635 640 Leu Asn Ile Gly Asn Ser Ala Leu Arg Gly Asn Phe Asn Gin Ala Phe 645 650 655 Ala Thr Ala Gly Val Ala Phe Leu Leu Glu Gly Phe Pro Glu Phe Thr 660 665 670 He Pro Ala Leu Gly val Phe Thr Phe Tyr Ser Ser Ile Gin Glu Arg 675 680 685 Glu Lys He lie Lys Thr He Glu Asn Cys Leu Glu Gin Arg val Lys 690 695 700 Arg Trp Lys Asp Ser Tyr Gin Trp Met val ser Asn Trp Leu ser Arg 705 710 715 720 ile Thr Thr Gin Phe Asn Hi s lie Asn Tyr Gin Met Tyr Asp ser Leu 725 730 735 ser Tyr Gin Ala Asp Ala lie Lys Ala Lys Ile Asp Leu Glu Tyr Lys 740 745 750 Lys Tyr Ser Gly Ser Asp Lys Glu Asn Ile Lys ser Gin Val Glu Asn 755 760 765 Leu Lys Asn Ser Leu Asp val Lys lie Ser Glu Ala Met Asn Asn lie 770 775 780 Asn Lys Phe ile Arg Gl u cys Ser val Thr Tyr Leu Phe Lys Asn Met 785 790 795 800 Leu Pro Lys val lie Asp Glu Leu Asn Lys Phe Asp Leu Arg Thr Lys 805 810 815 Thr Glu Leu lie Asn Leu ile Asp Ser Hi s Asn lie lie Leu Val Gly 820 825 830 Glu Val Asp Arg Leu Lys Ala Lys val Asn Glu Ser Phe Glu Asn Thr 835 840 845

    page 67

    P31003au-pct.txt

    2015268754 16 Dec 2015

    Met Pro Phe Asn lie Phe Ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp 850 855 860 ile lie Asn Glu Tyr phe Asn Leu Glu Gly Gly Gly Gly ser Gly Gly 865 870 875 880 Gly Gly Ser Gly Gly Gly Gly ser Ala Leu Val Gly Asn Hi s Trp Ala

    885 890 895

    Val Gly His Leu Met 900 <210> 26 <211> 899 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 26

    pro Val Thr lie Asn Asn Phe Asn Tyr Asn ASp Pro lie Asp Asn 15 Asn 1 5 10 Asn Ile lie Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg Tyr 20 25 30 Tyr Lys Ala Phe Lys Ile Thr Asp Arg Ile Trp ile Ile Pro Glu Arg 35 40 45 Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser ser Gly ile 50 55 60 Phe Asn Arg Asp Val Cys Gl u Tyr Tyr Asp Pro ASp Tyr Leu Asn Thr 65 70 75 80 Asn Asp Lys Lys Asn 85 Ile Phe Leu Gin Thr 90 Met lie Lys Leu Phe 95 Asn Arg Ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met ile lie 100 105 110

    Asn Gly lie Pro Tyr 115 Leu Gly Asp 120 Arg Arg Val Pro Leu 125 Glu Glu Phe Asn Thr Asn lie Ala Ser Val Thr val Asn Lys Leu Ile Ser Asn Pro 130 135 140 Gly Glu val Glu Arg Lys Lys Gly lie Phe Ala Asn Leu lie Ile Phe 145 150 155 160

    Page 68

    2015268754 16 Dec 2015

    Gly Pro Gly Pro val 165 P31003au-pct. txt lie Leu Asn Glu Asn Glu 170 Thr lie Asp He Gly 175 Gin Asn Hi s Phe Ala Ser Arg Glu Gly Phe Gly Gly lie Met Gin Met 180 185 190 Lys Phe Cys Pro Glu Tyr Val Ser Val Phe Asn Asn Val Gin Glu Asn 195 200 205 Lys Gly Ala Ser lie Phe Asn Arg Arg Gly Tyr Phe Ser Asp pro Ala 210 215 220 Leu lie Leu Met His Glu Leu lie His val Leu Hi s Gly Leu Tyr Gly 225 230 235 240 lie Lys Val Asp Asp Leu Pro lie Val Pro Asn Glu Lys Lys Phe Phe 245 250 255 Met Gin Ser Thr Asp Ala lie Gin Ala Glu Glu Leu Tyr Thr Phe Gly 260 265 270 Gly Gin Asp Pro Ser He lie Thr Pro Ser Thr Asp Lys Ser lie Tyr 275 280 285 Asp Lys Val Leu Gin Asn phe Arg Gly He val Asp Arg Leu Asn Lys 290 295 300 val Leu val cys He Ser Asp Pro Asn He Asn lie Asn He Tyr Lys 305 310 315 320 Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly Lys 325 330 335 Tyr Ser lie Asp Val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu Met 340 345 350 Phe Gly Phe Thr Glu Thr Asn lie Ala Glu Asn Tyr Lys lie Lys Thr 355 360 365 Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro val Lys He Lys Asn 370 375 380 Leu Leu Asp Asn Glu lie Tyr Thr lie Glu Glu Gly Phe Asn lie Ser 385 390 395 400 ASP Lys Asp Met Glu Lys Glu Tyr Arg Gly Gin Asn Lys Ala lie Asn 405 410 415 Lys Gin Ala Tyr Glu Glu lie Ser Lys Glu Hi s Leu Ala val Tyr Lys 420 425 430

    Page 69

    P31OO3au-pct.txt _J rle Gin Met Cys val Asp Glu Glu Lys Leu Tyr Asp Asp Asp Asp Lys _o 435 440 445 <N

    4sp Arg Trp Gly ser Ala ser Leu Gin Cys Ile Asp Val Asp Asn Glu Asp Q Leu 465 450 Phe Asp 470 455 460 Phe Ile Lys Asn Ser phe Ser 475 Asp Asp Leu Ser Lys 480 Asn Glu Arg Ile Glu Tyr Asn Thr Gin ser Asn Tyr ile Glu Asn Asp IT) oo 485 490 495 Phe Pro Ile Asn Glu Leu lie Leu Asp Thr Asp Leu ile Ser Lys lie 500 505 510 IT) o Glu Leu Pro Ser Glu Asn Thr Glu Ser Leu Thr Asp Phe Asn val Asp (N 515 520 525 val Pro val Tyr Glu Lys Gin Pro Ala lie Lys Lys ile Phe Thr Asp 530 535 540 Glu Asn Thr Ile Phe Gin Tyr Leu Tyr Ser Gin Thr phe Pro Leu Asp 545 550 555 560 lie Arg Asp lie ser Leu Thr Ser Ser phe ASp Asp Ala Leu Leu Phe 565 570 575 Ser Asn Lys val Tyr Ser Phe phe Ser Met Asp Tyr ile Lys Thr Ala 580 585 590 Asn Lys Val Val Glu Ala Gly Leu Phe Ala Gly Trp Val Lys Gin lie 595 600 605 val Asn Asp Phe val lie Glu Ala Asn Lys Ser Asn Thr Met Asp Ala 610 615 620 Ile Ala Asp lie Ser Leu ile val Pro Tyr Ile Gly Leu Ala Leu Asn 625 630 635 640 Val Gly Asn Glu Thr Ala Lys Gly Asn Phe Glu Asn Ala Phe Glu Ile 645 650 655 Al a Gly Ala Ser Ile Leu Leu Glu Phe ile Pro Glu Leu Leu lie Pro 660 665 670 Val Val Gly Ala Phe Leu Leu Glu Ser Tyr ile Asp Asn Lys Asn Lys 675 680 685 Ile lie Lys Thr lie Asp Asn Ala Leu Thr Lys Arg Asn Glu Lys Trp 690 695 700

    Page 70

    2015268754 16 Dec 2015

    Ser 705 Asp Met Tyr Gly P31OO3au-pct.txt Asn 720 Leu 710 lie val Ala Gln Trp 715 Leu Ser Thr Val Thr Gln Phe Tyr Thr Ile Lys Glu Gly Met Tyr Lys Ala Leu Asn Tyr 725 730 735 Gln Ala Gln Ala Leu Glu Glu lie lie Lys Tyr Arg Tyr Asn lie Tyr 740 745 750 Ser Glu Lys Glu Lys Ser Asn Ile Asn lie Asp Phe Asn Asp lie Asn 755 760 765 Ser Lys Leu Asn Glu Gly Ile Asn Gln Ala Ile Asp Asn lie Asn Asn 770 775 780 Phe Ile Asn Gly Cys Ser val ser Tyr Leu Met Lys Lys Met Ile Pro 785 790 795 800 Leu Ala val Glu Lys Leu Leu Asp Phe Asp Asn Thr Leu Lys Lys Asn 805 810 815 Leu Leu Asn Tyr Ile Asp Glu Asn Lys Leu Tyr Leu lie Gly Ser Ala 820 825 830 Glu Tyr Glu Lys ser Lys Val Asn Lys Tyr Leu Lys Thr lie Met Pro 835 840 845 Phe Asp Leu Ser lie Tyr Thr Asn Asp Thr He Leu lie Glu Met Phe 850 855 860 Asn Lys Tyr Asn Ser Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly 865 870 875 880 ser Gly Gly Gly Gly ser Ala Leu val Gly Asn Hi s Trp Ala Val Gly 885 890 895

    His Leu Met <210> 27 <211> 1065 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypepti de <400> 27

    Pro lie Thr Ile Asn Asn Phe Asn Tyr Ser Asp Pro val Asp Asn Lys 1 5 10 15 Asn ile Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu Pro

    Page 71

    P31OO3au-pct. txt

    20 25 30

    O

    ^Glu o Ω h ^phe r—H Lys Ala Phe Arg lie Thr Gly Asn lie Trp val pro 60 lie Pro Asp Arg 45 Ser 50 35 ser Asn pro 55 40 Arg Asn Pro Arg Val Thr Asn Leu Asn Lys ser Pro Lys Ser Gly Tyr Tyr Asp pro Asn Tyr Leu Ser Thr Asp Ser =t 65 70 75 80 υη oo Asp Lys Asp Thr Phe Leu Lys Glu lie lie Lys Leu Phe Lys Arg lie NO 85 90 95 ΓΊ 1 Asn Ser Arg Glu lie Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr Asp o 100 105 110 ΓΊ Ile Pro Phe Pro Gly Asn Asn Asn Thr pro Ile Asn Thr Phe Asp Phe 115 120 125 Asp Val Asp phe Asn Ser val Asp val Lys Thr Arg Gln Gly Asn Asn 130 135 140 Trp val Lys Thr Gly Ser Ile Asn Pro ser Val Ile lie Thr Gly Pro 145 150 155 160 Arg Glu Asn Ile Ile Asp Pro Glu Thr ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Ala Leu Ser Ile Ile ser 180 185 190 Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp Val 195 200 205 Gly Glu Gly Arg Phe ser Lys Ser Glu Phe Cys Met Asp Pro ile Leu 210 215 220 lie Leu Met Hi s Glu Leu Asn Hi s Ala Met Hi s Asn Leu Tyr Gly lie 225 230 235 240 Ala lie Pro Asn Asp Gln Thr ile ser Ser Val Thr Ser Asn ile Phe 245 250 255 Tyr Ser Gln Tyr Asn val Lys Leu Glu Tyr Ala Glu He Tyr Ala Phe 260 265 270 Gly Gly Pro Thr lie Asp Leu Ile pro Lys ser Ala Arg Lys Tyr phe 275 280 285 Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser lie Ala Lys Arg Leu Asn

    Page 72

    2015268754 16 Dec 2015

    P31OO3au-pct.txt

    290 295 300

    ser lie Thr Thr Ala Asn Pro ser ser Phe Asn Lys Tyr lie Gly Glu 320 305 310 315 Tyr Lys Gln Lys Leu lie Arg Lys Tyr Arg Phe val val Glu ser ser 325 330 335 Gly Glu Val Thr Val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn Glu 340 345 350 Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys He Tyr Asn val 355 360 365 Gln Asn Arg Lys lie Tyr Leu ser Asn val Tyr Thr Pro val Thr Al a 370 375 380 Asn ile Leu Asp Asp Asn Val Tyr Asp He Gln Asn Gly Phe Asn lie 385 390 395 400 Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe Cys Val Asp Ala lie Asp Gly Arg Ser Leu Tyr Asn Lys 435 440 445 Thr Leu Gln Cys Arg Glu Leu Leu Val Lys Asn Thr Asp Leu Pro Phe 450 455 460 lie Gly Asp Ile Ser Asp Val Lys Thr Asp Ile Phe Leu Arg Lys Asp 465 470 475 480 ile Asn Glu Glu Thr Glu val lie Tyr Tyr Pro Asp Asn Val Ser val 485 490 495 Asp Gln Val Ile Leu Ser Lys Asn Thr Ser Glu Hi s Gly Gln Leu Asp 500 505 510 Leu Leu Tyr Pro ser lie Asp Ser Glu Ser Glu Ile Leu Pro Gly Glu 515 520 525 Asn Gln Val Phe Tyr Asp Asn Arg Thr Gln Asn Val Asp Tyr Leu Asn 530 535 540 Ser Tyr Tyr Tyr Leu Glu ser Gln Lys Leu Ser Asp Asn Val Glu Asp 545 550 555 560 Phe Thr Phe Thr Arg Ser lie Glu Glu Ala Leu Asp Asn ser Ala Lys

    Page 73

    575

    565

    IT) ι—Η

    Ο

    ΓΗ

    Ο <υ

    Ω

    P31OO3au-pct.txt

    570

    ΙΓ)

    Ο

    ΓΗ

    Ο

    ΓΗ

    val Tyr Thr Tyr Phe Pro Thr Leu Ala Asn Lys val Asn Ala Gly val 580 585 590 Gin Gly Gly Leu Phe Leu Met Trp Ala Asn Asp Val val Glu Asp Phe 595 600 605 Thr Thr Asn lie Leu Arg Lys Asp Thr Leu Asp Lys lie Ser Asp val 610 615 620 Ser Ala lie lie Pro Tyr lie Gly Pro Ala Leu Asn lie Ser Asn ser 625 630 635 640 Val Arg Arg Gly Asn 645 Phe Thr Glu Ala Phe 650 Ala Val Thr Gly val 655 Thr lie Leu Leu Glu Ala Phe Pro Glu Phe Thr lie Pro Ala Leu Gly Ala 660 665 670 Phe val lie Tyr Ser Lys val Gin Glu Arg Asn Glu lie lie Lys Thr 675 680 685 lie Asp Asn cys Leu Glu Gin Arg lie Lys Arg Trp Lys Asp Ser Tyr 690 695 700 Glu Trp Met Met Gly Thr Trp Leu Ser Arg lie lie Thr Gin Phe Asn 705 710 715 720 Asn lie Ser Tyr Gin Met Tyr Asp ser Leu Asn Tyr Gin Ala Gly Ala 725 730 735 lie Lys Ala Lys lie Asp Leu Glu Tyr Lys Lys Tyr Ser Gl y Ser Asp 740 745 750 Lys Glu Asn lie Lys Ser Gin val Glu Asn Leu Lys Asn ser Leu Asp 755 760 765 val Lys lie Ser Glu Ala Met Asn Asn lie Asn Lys Phe lie Arg Glu 770 775 780 Cys Ser Val Thr Tyr Leu phe Lys Asn Met Leu Pro Lys val lie Asp 785 790 795 800 Glu Leu Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu He Asn Leu 805 810 815 lie Asp Ser Hi s Asn lie rle Leu val Gly Glu val Asp Lys Leu Lys 820 825 830 Ala Lys val Asn Asn Ser Phe Gin Asn Thr lie Pro Phe Asn lie Phe

    Page 74

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    835 840 845 Ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp lie ile Asn Glu Tyr Phe 850 855 860 Asn Leu Glu Gly Gly Gly Gly ser Gly Gly Gly Gly Ser Gly Gly Gly 865 870 875 880 Gly ser Ala Leu Val Ser Pro Leu Pro Ile Thr pro val Asn Ala Thr 885 890 895 Cys Ala ile Arg His Pro cys His Asn Asn Leu Met Asn Gln lie Arg 900 905 910 ser Gln Leu Al a Gln Leu Asn Gly Ser Ala Asn Ala Leu Phe lie Leu 915 920 925 Tyr Tyr Thr Ala Gln Gly Glu Pro Phe Pro Asn Asn Leu Asp Lys Leu 930 935 940 Cys Gly Pro Asn val Thr ASP Phe Pro Pro Phe His Ala Asn Gly Thr 945 950 955 960 Glu Lys Ala Lys Leu Val Glu Leu Tyr Arg Ile val val Tyr Leu Gly 965 970 975 Thr Ser Leu Gly Asn lie Thr Arg Asp Gln Lys ile Leu Asn Pro Ser 980 985 990 Ala Leu Ser Leu His Ser Lys Leu Asn i Ala . Thr Ala Asc i ile Leu Arg 995 1000 1005 Gly Leu Leu Ser Asn val Leu Cys Arg Leu Cys Ser Lys Tyr His 1010 1015 1020 Val Gly Hi s Val Asp Val Thr Tyr Gly Pro Asp Thr ser Gly Lys 1025 1030 1035 Asp Val Phe Gln Lys Lys Lys Leu Gly Cys Gln Leu Leu Gly Lys 1040 1045 1050 Tyr Lys Gln lie lie Ala Val Leu Ala Gln Ala Phe 1055 1060 1065

    <210> 28 <211> 1060 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide

    Page 75

    IT)

    P31003au-pct. txt

    r-H C— <400> 28 ν—> CN Pro 1 val Thr lie Asn 5 Asn Phe Asn Tyr Asn 10 Asp Pro Ile Asp Asn 15 Asn <D CD Q Asn lie Ile Met Met Glu pro Pro Phe Ala Arg Gly Thr Gly Arg Tyr 20 25 30 Tyr Lys Ala Phe Lys Ile Thr Asp Arg ile Trp lie lie pro Glu Arg 35 40 45 xr i/D o Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly lie OO 50 55 60 CN i/D Phe Asn Arg Asp val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn Thr H 65 70 75 80 © CN Asn Asp Lys Lys Asn Ile Phe Leu Gln Thr Met lie Lys Leu Phe Asn 85 90 95 Arg lie Lys ser 100 Lys Pro Leu Gly Glu 105 Lys Leu Leu Glu Met 110 lie Ile Asn Gly lie Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu Phe 115 120 125 Asn Thr Asn lie Ala ser val Thr Val Asn Lys Leu lie Ser Asn Pro 130 135 140 Gly Glu val Glu Arg Lys Lys Gly lie Phe Ala Asn Leu lie ile Phe 145 150 155 160 Gly Pro Gly pro val Leu Asn Glu Asn Glu Thr lie ASp lie Gly Ile 165 170 175 Gln Asn His phe Ala Ser Arg Gl u Gly phe Gly Gly lie Met Gln Met 180 185 190 Lys Phe cys Pro Glu Tyr val Ser val Phe Asn Asn val Gln Glu Asn 195 200 205 Lys Gly Ala Ser lie Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro Ala 210 215 220 Leu Ile Leu Met His Glu Leu lie Hi s val Leu Hi s Gly Leu Tyr Gly 225 230 235 240 ile Lys val Asp Asp Leu Pro lie Val Pro Asn Glu Lys Lys Phe Phe 245 250 255 Met Gln Ser Thr ASp Ala ile Gln Ala Glu Glu Leu Tyr Thr Phe Gly

    260 265 270

    Page 76

    P31OO3au-pct.txt

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    Gly Gin Asp 275 Pro Ser lie lie Thr Pro Ser Thr Asp Lys Ser lie Tyr 280 285 Asp Lys 290 val Leu Gin Asn Phe 295 Arg Gly lie val Asp 300 Arg Leu Asn Lys val Leu val Cys lie Ser Asp Pro Asn lie Asn lie Asn lie Tyr Lys 305 310 315 320 Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly Lys 325 330 335 Tyr Ser lie Asp 340 Val Glu Ser Phe Asp 345 Lys Leu Tyr Lys Ser 350 Leu Met Phe Gly Phe Thr Glu Thr Asn lie Ala Glu Asn Tyr Lys lie Lys Thr 355 360 365 Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro val Lys lie Lys Asn 370 375 380 Leu Leu Asp Asn Glu lie Tyr Thr lie Glu Glu Gly Phe Asn lie ser 385 390 395 400 Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gin Asn Lys Ala lie Asn 405 410 415 Lys Gin Ala Tyr Glu Glu lie Ser Lys Glu His Leu Ala Val Tyr Lys

    420 425 430

    lie Gin Met cys val Asp Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn 435 440 445 Asp Asp Asp Asp Lys Ser Pro Leu Pro lie Thr Pro Val Asn Ala Thr 450 455 460 Cys Ala lie Arg Hi s Pro Cys His Asn Asn Leu Met Asn Gin lie Arg 465 470 475 480 ser Gin Leu Ala Gin Leu Asn Gly Ser Ala Asn Al a Leu Phe lie Leu 485 490 495 Tyr Tyr Thr Ala Gin Gly Glu Pro Phe Pro Asn Asn Leu Asp Lys Leu 500 505 510 Cys Gly Pro Asn Val Thr Asp Phe Pro Pro Phe Hi s Ala Asn Gly Thr 515 520 525 Glu Lys Ala Lys Leu val Glu Leu Tyr Arg lie val val Tyr Leu Gly 530 535 540

    Page 77 p31OO3au-pct.txt

    o _u ΓΗ rhr Ser Leu Gly Asn lie Thr Arg 550 Asp Gin Lys lie Leu 555 Asn Pro ser 560 Ο Ο 545 Ω Ala Leu Ser Leu Hi s Ser Lys Leu Asn Ala Thr Ala Asp Ile Leu Arg 565 570 575 Gly Leu Leu Ser Asn val Leu cys Arg Leu cys ser Lys Tyr Hi s val 580 585 590 ^ι- ιη ο Gly His Val Asp Val Thr Tyr Gly pro Asp Thr ser Gly Lys Asp val 00 (Μ QD 595 600 605 Phe Gin Lys Lys Lys Leu Gly Cys Gin Leu Leu Gly Lys Tyr Lys Gin Ο Γ4 610 615 620 Ile lie Ala val Leu Ala Gin Ala Phe Ala Glu Ala Ala Ala Lys Glu 625 630 635 640 Ala Ala Ala Lys Ala Leu Gin cys lie Asp val Asp Asn Glu Asp Leu 645 650 655 Phe Phe ile Ala Asp Lys Asn Ser Phe Ser Asp Asp Leu Ser Lys Asn 660 665 670 Glu Arg lie Glu Tyr Asn Thr Gin ser Asn Tyr Ile Glu Asn Asp Phe 675 680 685 Pro ile Asn Glu Leu ile Leu ASp Thr Asp Leu lie ser Lys lie Glu 690 695 700 Leu Pro ser Glu Asn Thr Glu Ser Leu Thr Asp phe Asn val Asp val 705 710 715 720 Pro val Tyr Glu Lys Gin Pro Ala lie Lys Lys lie Phe Thr Asp Glu 725 730 735 Asn Thr lie Phe Gin Tyr Leu Tyr Ser Gin Thr Phe Pro Leu Asp lie 740 745 750 Arg Asp lie ser Leu Thr ser Ser Phe Asp Asp Ala Leu Leu Phe Ser 755 760 765 Asn Lys Val Tyr Ser Phe Phe Ser Met Asp Tyr lie Lys Thr Ala Asn 770 775 780 Lys val Val Glu Ala Gly Leu Phe Ala Gly Trp val Lys Gin lie Val 785 790 795 800 Asn Asp Phe val lie Glu Ala Asn Lys Ser Asn Thr Met Asp Lys ile 805 810 815

    Page 78

    P31OO3au-pct.txt

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    Ala Asp Ile Ser Leu Ile Val Pro Tyr ile Gly 825 Leu Ala Leu Asn Val 830 820 Gly Asn Glu Thr Ala Lys Gly Asn Phe Glu Asn Ala Phe Glu Ile Ala 835 840 845 Gly Ala Ser lie Leu Leu Glu Phe lie Pro Glu Leu Leu lie Pro val 850 855 860 Val Gly Ala Phe Leu Leu Glu Ser Tyr ile Asp Asn Lys Asn Lys Ile 865 870 875 880 lie Lys Thr lie Asp Asn Ala Leu Thr Lys Arg Asn Glu Lys Trp ser 885 890 895 Asp Met Tyr Gly Leu Ile val Ala Gln Trp Leu Ser Thr Val Asn Thr 900 905 910 Gln Phe Tyr Thr lie Lys Glu Gly Met Tyr Lys Ala Leu Asn Tyr Gln 915 920 925 Ala Gln Ala Leu Glu Glu Ile ile Lys Tyr Arg Tyr Asn Ile Tyr Ser 930 935 940 Glu Lys Glu Lys Ser Asn lie Asn lie Asp Phe Asn Asp Ile Asn Ser 945 950 955 960 Lys Leu Asn Glu Gly Ile Asn Gln Ala Ile Asp Asn lie Asn Asn Phe 965 970 975 lie Asn Gly cys ser Val ser Tyr Leu Met Lys Lys Met Ile Pro Leu 980 985 990 Ala Val Glu Lys Leu Leu Asp Phe Asp Asn Thr ' Leu Lys ; Lys Asn Leu 995 1000 1005 Leu Asn Tyr ile Asp Glu Asn Lys Leu Tyr Leu lie Gly Ser Ala 1010 1015 1020 Glu Tyr Glu Lys Ser Lys Val Asn Lys Tyr Leu Lys Thr lie Met 1025 1030 1035 Pro Phe Asp Leu Ser lie Tyr Thr Asn Asp Thr Ile Leu lie Glu 1040 1045 1050

    <210> 29 <211> 1026

    Page 79

    Met Phe Asn Lys Tyr Asn Ser

    1055 1060

    P31OO3au-pct.txt

    6 Dec 2015 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 29 on

    ΓΟΟ

    CH un o

    OH

    Pro lie Thr 1 ile Asn 5 Asn Phe Asn Tyr ser Asp 10 Pro val Asp Asn 15 Lys Asn lie Leu Tyr Leu Asp Thr His Leu Asn Thr Leu Ala Asn Glu Pro 20 25 30 Gl u Lys Ala Phe Arg lie Thr Gly Asn lie Trp val ile Pro Asp Arg 35 40 45 Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg val Thr 50 55 60 Ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp ser 65 70 75 80 Asp Lys Asp Thr Phe Leu Lys Glu Ile Ile Lys Leu Phe Lys Arg Ile 85 90 95 Asn Ser Arg Glu lie Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr Asp 100 105 110 lie Pro Phe Pro Gly Asn Asn Asn Thr Pro lie Asn Thr Phe Asp Phe 115 120 125 Asp Val Asp Phe Asn Ser Val Asp Val Lys Thr Arg Gin Gly Asn Asn 130 135 140 Trp val Lys Thr Gly Ser Ile Asn Pro ser val Ile Ile Thr Gly Pro 145 150 155 160 Arg Glu Asn Ile Ile Asp Pro Glu Thr ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Ala Ala Gin Glu Gly Phe Gly Ala Leu Ser lie Ile Ser 180 185 190 Ile Ser Pro Arg Phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp Val 195 200 205 Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe cys Met Asp Pro lie Leu 210 215 220 ile Leu Met Hi s Glu Leu Asn His Ala Met His Asn Leu Tyr Gly Ile

    225 230 235 240

    Page 80

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Ala Ile Pro Asn Asp Gin Thr lie Ser Ser val Thr Ser Asn lie Phe 245 250 255 Tyr Ser Gin Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala Phe 260 265 270 Gly Gly Pro Thr Ile Asp Leu lie Pro Lys ser Ala Arg Lys Tyr Phe 275 280 285 Glu Glu 290 Lys Ala Leu Asp Tyr 295 Tyr Arg Ser ile Ala 300 Lys Arg Leu Asn Ser He Thr Thr Ala Asn Pro ser Ser Phe Asn Lys Tyr Ile Gly Glu 305 310 315 320 Tyr Lys Gin Lys Leu lie Arg Lys Tyr Arg Phe val Val Glu Ser Ser 325 330 335 Gly Glu val Thr Val Asn Arg Asn Lys Phe val Glu Leu Tyr Asn Glu 340 345 350 Leu Thr Gin Ile Phe Thr Glu Phe Asn Tyr Ala Lys Ile Tyr Asn val 355 360 365 Gin Asn Arg Lys Ile Tyr Leu ser Asn val Tyr Thr Pro val Thr Ala 370 375 380 Asn Ile Leu Asp Asp Asn Val Tyr Asp lie Gin Asn Gly Phe Asn He 385 390 395 400 Pro Lys Ser Asn Leu Asn val Leu Phe Met Gly Gin Asn Leu Ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe Cys val Asp Ala lie Asp Gly Arg Ser Leu Tyr Asn Lys 435 440 445 Thr Leu Gl n Cys Arg Glu Leu Leu Val Lys Asn Thr Asp Leu Pro Phe 450 455 460 lie Gly Asp Ile ser Asp Val Lys Thr Asp lie Phe Leu Arg Lys Asp 465 470 475 480 Ile Asn Glu Glu Thr Glu val lie Tyr Tyr Pro Asp Asn Val Ser Val 485 490 495 Asp Gin val lie Leu Ser Lys Asn Thr Ser Glu His Gly Gin Leu Asp 500 505 510

    Page 81 in

    P31003au-pct.txt

    o ^Leu CM Leu Tyr Pro ser lie Asp Ser Glu ser Glu lie Leu Pro Gly Glu 515 520 Asn 525 Asn o CD Ω Asn Gin val 530 Phe Tyr Asp Asn 535 Arg Thr Gin Val 540 ASp Tyr Leu Ser 545 Tyr Tyr Tyr Leu Glu 550 Ser Gin Lys Leu Ser 555 Asp Asn val Glu Asp 560 in Phe Thr Phe Thr Arg 565 Ser lie Glu Glu Ala 570 Leu Asp Asn Ser Ala 575 Lys oo CM in val Tyr Thr Tyr Phe 580 Pro Thr Leu Ala 585 Asn Lys val Asn Al a 590 Gly val CM Gin Gl y Gl y 595 Leu Phe Leu Met Trp Ala 600 Asn Asp val val 605 Glu Asp Phe Thr Thr Asn 610 lie Leu Arg Lys 615 Asp Thr Leu Asp Lys 620 lie Ser Asp val Ser 625 Ala lie lie Pro Tyr 630 He Gly Pro Ala Leu 635 Asn lie Ser Asn Ser 640 Val Arg Arg Gly Asn 645 Phe Thr Glu Ala Phe 650 Ala val Thr Gly Val 655 Thr lie Leu Leu Glu Ala 660 Phe Pro Glu Phe 665 Thr lie pro Ala Leu 670 Gly Ala Phe val lie 675 Tyr ser Lys Val Gl n Gl u 680 Arg Asn Glu lie 685 lie Lys Thr lie Asp Asn 690 cys Leu Glu Gin 695 Arg lie Lys Arg Trp 700 Lys Asp Ser Tyr Glu 705 Trp Met Met Gly Thr 710 Trp Leu ser Arg lie 715 lie Thr Gin Phe Asn 720 Asn lie ser Tyr Gin 725 Met Tyr Asp Ser Leu 730 Asn Tyr Gin Ala Gly 735 Ala lie Lys Ala Lys lie 740 Asp Leu Glu Tyr 745 Lys Lys Tyr Ser Gly 750 Ser Asp Lys Glu Asn 755 lie Lys ser Gin Val Glu 760 Asn Leu Lys Asn 765 Ser Leu Asp val Lys lie 770 Ser Glu Ala Met 775 Asn Asn lie Asn Lys 780 Phe lie Arg Glu

    Page 82

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    cys Ser val 785 Thr Tyr Leu Phe Lys 790 Asn Met Leu Pro Lys 795 val Ile Asp 800 Glu Leu Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu lie Asn Leu 805 810 815 lie Asp Ser His Asn lie lie Leu Val Gly Glu Val Asp Lys Leu Lys 820 825 830 Ala Lys val Asn Asn Ser Phe Gin Asn Thr Ile Pro Phe Asn Ile Phe 835 840 845 Ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp Ile Ile Asn Glu Tyr Phe 850 855 860

    Asn Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 865 870 875 880

    Gly Ser Ala Leu val 885 Met Phe Asn Leu Pro Pro Gly 890 Asn Tyr Lys 895 Lys Pro Lys Leu Leu Tyr cys Ser Asn Gly Gly His Phe Leu Arg Ile Leu 900 905 910 Pro Asp Gly Thr val Asp Gly Thr Arg Asp Arg Ser Asp Gin His lie 915 920 925 Gin Leu Gin Leu Ser Ala Glu Ser val Gly Glu val Tyr lie Lys Ser 930 935 940 Thr Glu Thr Gly Gin Tyr Leu Ala Met Asp Thr Asp Gly Leu Leu Tyr 945 950 955 960 ( Gly Ser Gin Thr Pro Asn Glu Glu cys Leu Phe Leu Glu Arg Leu Glu 965 970 975 Glu Asn His Tyr Asn Thr Tyr Ile Ser Lys Lys Hi s Ala Glu Lys Asn

    980 985 990

    Trp Phe val Gly Leu Lys Lys Asn Gly ser Cys Lys Arg Gly Pro Arg 995 1000 1005

    Thr His Tyr Gly Gin Lys Ala Ile Leu Phe Leu Pro Leu Pro val 1010 1015 1020

    Ser Ser Asp 1025 <210> 30 <211> 1018 <212> PRT

    Page 83

    P31OO3au-pct. txt <213> Artificial Sequence

    S <220>

    <221> source

    Q <223> /note=Description of Artificial sequence: Synthetic polypeptide

    Q _ <400> 30

    Glu Phe val Asn 1 Lys 5 Gin Phe Asn Tyr Lys Asp Pro Val Asn Gly val 10 15 tt)^- ASp Ile Ala Tyr Ile Lys Ile Pro Asn Ala Gly Gin Met Gin Pro val QD 20 25 30 O 00 Lys Ala Phe Lys Ile Hi s Asn Lys Ile Trp val Ile Pro Glu Arg Asp n 35 40 45 QD Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala n 50 55 60 Lys Gin Val Pro Val Ser Tyr Tyr Asp ser Thr Tyr Leu Ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly val Thr Lys Leu Phe Glu Arg 85 90 95 lie Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr ser lie val Arg 100 105 110 Gly Ile Pro Phe Trp Gl y Gl y Ser Thr ile Asp Thr Glu Leu Lys val 115 120 125 Ile Asp Thr Asn Cys lie Asn Val Ile Gin Pro Asp Gly Ser Tyr Arg 130 135 140 ser Glu Glu Leu Asn Leu Val Ile Ile Gly Pro ser Ala Asp Ile Ile 145 150 155 160 Gin Phe Glu Cys Leu ser Phe Gly His Gl u Val Leu Asn Leu Thr Arg 165 170 175 Asn Gly Tyr Gly Ser Thr Gin Tyr ile Arg Phe ser Pro Asp Phe Thr 180 185 190 Phe Gly Phe Glu Glu Ser Leu Glu val Asp Thr Asn Pro Leu Leu Gly 195 200 205 Ala Gly Lys Phe Ala Thr Asp Pro Ala val Thr Leu Ala Hi s Glu Leu 210 215 220 Ile Hi s Ala Gly Hi s Arg Leu Tyr Gly lie Ala Ile Asn Pro Asn Arg 225 230 235 240

    Page 84

    2015268754 16 Dec 2015

    val Phe Lys val P31OO3au-pct.txt Asn 245 Thr Asn Ala Tyr Tyr 250 Glu Met Ser Gly Leu 255 Glu val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe 260 265 270 lie Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys 275 280 285 Phe Lys Asp lie Ala Ser Thr Leu Asn Lys Ala Lys Ser lie val Gly 290 295 300 Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu lie Tyr Thr Glu ASp Asn 340 345 350 Phe val Lys Phe Phe Lys val Leu Asn Arg Lys Thr Tyr Leu Asn Phe 355 360 365 Asp Lys Ala val Phe Lys lie Asn lie Val Pro Lys val Asn Tyr Thr 370 375 380 lie Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe 385 390 395 400 Asn Gly Gin Asn Thr Glu lie Asn Asn Met Asn Phe Thr Lys Leu Lys 405 410 415 Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu cys Val Asp Gly 420 425 430 lie lie Thr Ser Lys Thr Lys Ser Asp Asp Asp Asp Lys Met Phe Asn 435 440 445 Leu Pro Pro Gly Asn Tyr Lys Lys Pro Lys Leu Leu Tyr cys Ser Asn 450 455 460 Gly Gly Hi s Phe Leu Arg lie Leu Pro Asp Gly Thr Val Asp Gly Thr 465 470 475 480 Arg Asp Arg Ser Asp Gin His lie Gin Leu Gin Leu Ser Ala Glu Ser 485 490 495 val Gly Glu val Tyr lie Lys Ser Thr Glu Thr Gly Gin Tyr Leu Ala 500 505 510

    Page 85

    IZ*\ P3H 0O3ai j-pc t.tx· t Met Asp Thr Asp Gly Leu Leu Tyr Gly Ser Gin Thr pro Asn Glu Glu o 515 520 525 <N <D cys Leu Phe Leu Glu Arg Leu Glu Glu Asn Hi s Tyr Asn Thr Tyr lie CD 530 535 540 Q Ser Lys Lys Hi s Ala Glu Lys Asn Trp Phe Val Gly Leu Lys Lys Asn 545 550 555 560 τ-b Gly Ser cys Lys Arg Gly pro Arg Thr Hi s Tyr Gly Gin Lys Ala He \l in 565 570 575 O oo Leu Phe Leu Pro Leu pro val Ser Ser Asp Gly Gly Gly Gly Ser Gly CH 580 585 590 in Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu val Leu Gin Cys lie <N 595 600 605 Lys Val Asn Asn Trp Asp Leu Phe Phe Ser Pro ser Glu Asp Asn Phe 610 615 620 Thr Asn Asp Leu Asn Lys Gly Glu Gl u lie Thr Ser Asp Thr Asn lie 625 630 635 640 Glu Ala Ala Glu Glu Asn lie Ser Leu Asp Leu lie Gin Gin Tyr Tyr 645 650 655 Leu Thr Phe Asn Phe Asp Asn Glu Pro Glu Asn lie Ser lie Glu Asn 660 665 670 Leu ser Ser Asp lie lie Gly Gin Leu Glu Leu Met Pro Asn lie Glu 675 680 685 Arg Phe pro Asn Gly Lys Lys Tyr Glu Leu Asp Lys Tyr Thr Met Phe 690 695 700 His Tyr Leu Arg Ala Gin Glu Phe Glu His Gly Lys ser Arg lie Ala 705 710 715 720 Leu Thr Asn Ser val Asn Glu Ala Leu Leu Asn Pro Ser Arg val Tyr 725 730 735 Thr Phe Phe Ser Ser Asp Tyr val Lys Lys Val Asn Lys Ala Thr Glu 740 745 750 Ala Ala Met Phe Leu Gly Trp val Glu Gin Leu val Tyr Asp Phe Thr 755 760 765 Asp Glu Thr Ser Glu Val ser Thr Thr Asp Lys lie Ala Asp lie Thr 770 775 780

    Page 86

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Ile Ile lie Pro Tyr Ile Gly Pro Ala Leu Asn 795 ile Gly Asn Met Leu 800 785 790 Tyr Lys Asp ASp Phe val Gly Ala Leu Ile Phe Ser Gly Ala Val Ile 805 810 815 Leu Leu Glu Phe lie Pro Glu ile Ala Ile Pro Val Leu Gly Thr Phe 820 825 830 Ala Leu val Ser Tyr Ile Ala Asn Lys val Leu Thr val Gin Thr lie 835 840 845 Asp Asn Ala Leu Ser Lys Arg Asn Glu Lys Trp Asp Glu Val Tyr Lys 850 855 860 Tyr Ile val Thr Asn Trp Leu Ala Lys val Asn Thr Gin Ile Asp Leu 865 870 875 880 ile Arg Lys Lys Met Lys Glu Ala Leu Glu Asn Gin Ala Glu Ala Thr 885 890 895 Lys Ala Ile lie Asn Tyr Gin Tyr Asn Gin Tyr Thr Glu Glu Glu Lys 900 905 910 Asn Asn lie Asn Phe Asn Ile Asp Asp Leu Ser ser Lys Leu Asn Glu 915 920 925 Ser lie Asn Lys Ala Met lie Asn lie Asn Lys Phe Leu Asn Gin cys 930 935 940 Ser Val ser Tyr Leu Met Asn Ser Met Ile Pro Tyr Gly Val Lys Arg 945 950 955 960 Leu Glu Asp Phe Asp Ala Ser Leu Lys Asp Ala Leu Leu Lys Tyr ile 965 970 975 Tyr Asp Asn Arg Gly Thr Leu Ile Gly Gin val Asp Arg Leu Lys Asp 980 985 990 Lys val Asn Asn Thr Leu Ser Thr Asp 1 > lie ! Pre » Phe > Gin i Leu Ser Lys 995 100C 1005

    Tyr val Asp Asn Gin Arg Leu Leu Ser Thr 1010 1015 <210> 31 <211> 1063 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic Page 87

    P31OO3au-pct.txt iT) o

    o <D

    Q

    ΙΤ) o

    oo <N in o

    <N polypeptide <400> 31

    Glu Phe 1 val Asn Lys Gin Phe Asn Tyr Lys Asp Pro val Asn Gly 15 Val 5 10 Asp lie Ala Tyr lie Lys lie Pro Asn Ala Gly Gin Met Gin Pro val 20 25 30 Lys Ala Phe Lys lie His Asn Lys lie Trp Val lie Pro Glu Arg Asp 35 40 45 Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu Ala 50 55 60 Lys Gin val Pro val Ser Tyr Tyr Asp ser Thr Tyr Leu ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly val Thr Lys Leu Phe Glu Arg 85 90 95 lie Tyr Ser Thr ASp Leu Gly Arg Met Leu Leu Thr ser lie Val Arg 100 105 110 Gly lie Pro Phe Trp Gly Gly Ser Thr lie Asp Thr Glu Leu Lys val 115 120 125 lie ASp Thr Asn cys lie Asn Val lie Gin Pro Asp Gly Ser Tyr Arg 130 135 140 Ser Glu Glu Leu Asn Leu Val lie lie Gly Pro Ser Ala Asp lie lie 145 150 155 160 Gin Phe Glu Cys Leu ser Phe Gly Hi s Glu Val Leu Asn Leu Thr Arg 165 170 175 Asn Gly Tyr Gly Ser Thr Gin Tyr lie Arg Phe ser Pro Asp Phe Thr 180 185 190 Phe Gly Phe Glu Glu ser Leu Glu val Asp Thr Asn Pro Leu Leu Gly 195 200 205 Ala Gly Lys Phe Ala Thr Asp Pro Al a val Thr Leu Ala Hi s Glu Leu 210 215 220 lie His Ala Gly His Arg Leu Tyr Gly lie Ala lie Asn Pro Asn Arg 225 230 235 240 Val Phe Lys val Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu Glu

    245 250 255

    Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys Phe Page 88

    2015268754 16 Dec 2015

    260 P31003au-pct.txt 265 270 Ile Asp Ser Leu Gln Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys 275 280 285 Phe Lys Asp lie Ala ser Thr Leu Asn Lys Ala Lys Ser lie Val Gly 290 295 300 Thr Thr Ala Ser Leu Gln Tyr Met Lys Asn Val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe ser val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu lie Tyr Thr Glu Asp Asn 340 345 350 Phe val Lys Phe Phe Lys val Leu Asn Arg Lys Thr Tyr Leu Asn Phe 355 360 365 Asp Lys Ala val phe Lys lie Asn Ile Val Pro Lys val Asn Tyr Thr 370 375 380 Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe 385 390 395 400 Asn Gly Gln Asn Thr Glu lie Asn Asn Met Asn Phe Thr Lys Leu Lys 405 410 415 Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Asp Gly 420 425 430 lie Ile Thr Ser Lys Thr Lys Ser Asp ASp Asp Asp Lys Asn Lys Ala 435 440 445 Leu Asn Leu Gln cys Ile Lys val Asn Asn Trp Asp Leu Phe Phe Ser 450 455 460 Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu Ile 465 470 475 480 Thr Ser Asp Thr Asn lie Glu Ala Ala Glu Glu Asn Ile Ser Leu Asp 485 490 495 Leu lie Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro Glu 500 505 510 Asn lie Ser Ile Glu Asn Leu Ser Ser Asp Ile lie Gly Gln Leu Glu 515 520 525

    Leu Met Pro Asn Ile Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu Leu Page 89

    P31003au-pct.txt

    540

    530 535

    Ο

    CM Asp Lys Tyr Thr Arg Met Phe 550 His Leu Tyr Leu Arg Ala Gln Glu Phe Glu 555 Hi s 560 Leu S⁵⁴⁵ Q _Ί Gly Lys ser Ile 565 Ala Thr Asn ser 570 val Asn Glu Ala Leu 575 Asn Pro ser Arg val Tyr Thr Phe Phe ser Ser Asp Tyr Val Lys Lys IT) O OO 580 585 590 val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu Gln CM IT) 595 600 605 Leu val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr Asp o 610 615 620 CM Lys lie Ala Asp ile Thr ile lie lie Pro Tyr ile Gly Pro Ala Leu 625 630 635 640 Asn lie Gly Asn Met Leu Tyr Lys Asp Asp Phe val Gly Ala Leu Ile 645 650 655 Phe ser Gly Ala Val lie Leu Leu Glu Phe lie pro Gl u He Ala Ile 660 665 670 Pro val Leu Gly Thr Phe Ala Leu Val Ser Tyr lie Ala Asn Lys Val 675 680 685 Leu Thr val Gln Thr Ile Asp Asn Ala Leu Ser Lys Arg Asn Glu Lys 690 695 700 Trp Asp Glu Val Tyr Lys Tyr lie Val Thr Asn Trp Leu Ala Lys val 705 710 715 720 Asn Thr Gln lie Asp Leu lie Arg Lys Lys Met Lys Glu Al a Leu Glu 725 730 735 Asn Gln Ala Glu Ala Thr Lys Ala ile lie Asn Tyr Gln Tyr Asn Gln 740 745 750 Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn lie Asp Asp Leu 755 760 765 Ser ser Lys Leu Asn Glu Ser Ile Asn Lys Ala Met Ile Asn lie Asn 770 775 780 Lys Phe Leu Asn Gln Cys Ser Val ser Tyr Leu Met Asn Ser Met lie 785 790 795 800 Pro Tyr Gly val Lys Arg Leu Glu Asp Phe Asp Al a Ser Leu Lys Asp

    Page 90

    P31003au-pct.txt

    2015268754 16 Dec 2015

    805 810 815 Ala Leu Leu Lys Tyr ile Tyr Asp Asn Arg Gly Thr Leu Ile Gly Gin 820 825 830 val Asp Arg Leu Lys ASp Lys val Asn Asn Thr Leu Ser Thr Asp lie 835 840 845 Pro Phe Gin Leu Ser Lys Tyr Val Asp Asn Gin Arg Leu Leu Ser Thr 850 855 860 Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 865 870 875 880 Ser Gly Gly Gly Gly Ser Ala Leu val Asp His Leu Gly Gin Ser Glu 885 890 895 Ala Gly Gly Leu 900 Pro Arg Gly Pro Ala 905 val Thr Asp Leu Asp 910 His Leu Lys Gly Ile Leu Arg Arg Arg Gin Leu Tyr Cys Arg Thr Gly Phe Hi s 915 920 925 Leu Glu Ile Phe Pro Asn Gly Thr Ile Gin Gly Thr Arg Lys Asp Hi s 930 935 940 Ser Arg Phe Gly lie Leu Glu Phe lie ser lie Ala Val Gly Leu val 945 950 955 960 Ser Ile Arg Gly Val Asp Ser Gly Leu Tyr Leu Gly Met Asn Glu Lys 965 970 975 Gly Glu Leu Tyr Gly ser Glu Lys Leu Thr Gin Glu Cys Val Phe Arg 980 985 990 Glu Gin Phe Glu Glu Asn Trp Tyr Asn i Thr Tyr ¹ Ser ¹ Ser As ;n Le ;u Tyr

    995 1000 1005

    Lys His val Asp Thr Gly Arg Arg Tyr Tyr Val Ala Leu Asn Lys 1010 1015 1020

    Asp Gly Thr Pro Arg Glu Gly Thr Arg Thr Lys Arg His Gin Lys 1025 1030 1035

    Phe Thr His Phe Leu pro Arg Pro Val Asp Pro Asp Lys Val Pro 1040 1045 1050

    Glu Leu Tyr Lys Asp lie Leu ser Gin ser 1055 1060 <210> 32

    Page 91

    P31003au-pct. txt

    ZJ <211> 1055 _o <212> PRT <213> Artificial Sequence

    O <220>

    0) <221> source

    Q <223> /note=Description of Artificial sequence: synthetic polypeptide <400> 32

    Pro lie Thr lie Asn Asn Phe Asn Tyr ser Asp Pro Val Asp Asn 15 Lys 1 5 10 1 V) Asn Ile Leu Tyr Leu ASp Thr His Leu Asn Thr Leu Ala Asn Glu Pro OO 20 25 30 kO CN Glu Lys Ala Phe Arg lie Thr Gly Asn Ile Trp val lie Pro Asp Arg 35 40 45 o CN Phe Ser Arg Asn Ser Asn Pro Asn Leu Asn Lys Pro Pro Arg Val Thr 50 55 60 ser Pro Lys Ser Gly Tyr Tyr Asp Pro Asn Tyr Leu Ser Thr Asp ser 65 70 75 80 Asp Lys Asp Thr Phe Leu Lys Glu Ile lie Lys Leu Phe Lys Arg lie 85 90 95 Asn ser Arg Glu Ile Gly Glu Glu Leu Ile Tyr Arg Leu Ser Thr Asp 100 105 110 Ile Pro Phe Pro Gly Asn Asn Asn Thr Pro lie Asn Thr phe Asp Phe 115 120 125 Asp val Asp Phe Asn Ser Val ASp Val Lys Thr Arg Gln Gly Asn Asn 130 135 140 Trp Val Lys Thr Gly Ser Ile Asn Pro Ser Val Ile Ile Thr Gly Pro 145 150 155 160 Arg Glu Asn lie Ile Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Al a Ala Gln Glu Gly Phe Gly Ala Leu Ser lie Ile ser 180 185 190 Ile ser Pro Arg Phe Met Leu Thr Tyr ser Asn Ala Thr Asn Asp val 195 200 205 Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe cys Met Asp Pro Ile Leu 210 215 220 ile Leu Met Hi s Glu Leu Asn Hi s Ala Met His Asn Leu Tyr Gly Ile 225 230 235 240

    Page 92

    2015268754 16 Dec 2015

    P31OO3au~pct.txt

    Ala lie Pro Asn Asp Gin Thr lie ser Ser val Thr Ser Asn lie 255 Phe 245 250 Tyr ser Gin Tyr Asn val Lys Leu Glu Tyr Ala Glu lie Tyr Ala Phe 260 265 270 Gly Gly Pro Thr lie Asp Leu lie Pro Lys Ser Ala Arg Lys Tyr Phe 275 280 285 Glu Glu 290 Lys Ala Leu Asp Tyr 295 Tyr Arg Ser lie Ala 300 Lys Arg Leu Asn Ser lie Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr lie Gly Glu 305 310 315 320 Tyr Lys Gin Lys Leu lie Arg Lys Tyr Arg Phe Val val Glu Ser ser 325 330 335 Gly Glu val Thr val Asn Arg Asn Lys Phe val Glu Leu Tyr Asn Glu 340 345 350 Leu Thr Gin lie Phe Thr Glu Phe Asn Tyr Ala Lys lie Tyr Asn val 355 360 365 Gin Asn Arg Lys lie Tyr Leu Ser Asn val Tyr Thr Pro val Thr Ala 370 375 380 Asn lie Leu Asp Asp Asn Val Tyr Asp lie Gin Asn Gly Phe Asn lie 385 390 395 400 Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gin Asn Leu Ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys Val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe cys Val Asp Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn 435 440 445 Asp Asp Asp Asp Lys Asp His Leu Gly Gin Ser Glu Ala Gly Gly Leu 450 455 460 Pro Arg Gly Pro Ala val Thr Asp Leu Asp Hi s Leu Lys Gly lie Leu 465 470 475 480 Arg Arg Arg Gin Leu Tyr Cys Arg Thr Gly Phe His Leu Glu lie Phe 485 490 495 Pro Asn Gly Thr lie Gin Gly Thr Arg Lys Asp Hi s Ser Arg phe Gly 500 505 510

    Page 93

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    IT)

    S ile Leu Glu Phe Ile Ser Ile Ala val Gly Leu val Ser 525 Ile Arg Gly O <D 515 520 Ω Val Asp Ser Gly Leu Tyr Leu Gly Met Asn Glu Lys Gly Glu Leu Tyr 530 535 540 Gly Ser Glu Lys Leu Thr Gln Glu cys Val Phe Arg Glu Gln phe Glu 545 550 555 560 ^1^- MD Thr His val Glu Asn Trp Tyr Asn Tyr Ser Ser Asn Leu Tyr Lys Asp oo kO CH MD 565 570 575 Thr Gly Arg Arg Tyr Tyr Val Ala Leu Asn Lys Asp Gly Thr Pro Arg o <N 580 585 590 Glu Gly Thr Arg Thr Lys Arg Hi s Gln Lys Phe Thr Hi s Phe Leu Pro 595 600 605 Arg Pro val Asp Pro ASp Lys val pro Glu Leu Tyr Lys Asp lie Leu 610 615 620 Ser Gln Ser Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Leu 625 630 635 640 Gln cys Arg Glu Leu Leu val Lys Asn Thr Asp Leu Pro Phe lie Gly 645 650 655 Asp Ile ser Asp val Lys Thr Asp lie Phe Leu Arg Lys Asp ile Asn 660 665 670 Glu Glu Thr Glu Val lie Tyr Tyr Pro Asp Asn val Ser Val Asp Gln 675 680 685 Val Ile Leu ser Lys Asn Thr Ser Glu His Gly Gln Leu Asp Leu Leu 690 695 700 Tyr Pro ser lie Asp ser Glu Ser Glu lie Leu Pro Gly Glu Asn Gln 705 710 715 720 Val Phe Tyr Asp Asn Arg Thr Gln Asn val Asp Tyr Leu Asn Ser Tyr 725 730 735 Tyr Tyr Leu Glu Ser Gln Lys Leu Ser Asp Asn val Glu Asp Phe Thr 740 745 750 Phe Thr Arg ser He Glu Glu Ala Leu Asp Asn Ser Ala Lys val Tyr 755 760 765 Thr Tyr Phe Pro Thr Leu Ala Asn Lys Val Asn Ala Gly val Gln Gly 770 775 780

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    Gly Leu Phe Leu Met 785 Trp 790 Ala Asn Asp val val 795 Glu Asp Phe Thr Thr 800 Asn lie Leu Arg Lys Asp Thr Leu Asp Lys lie Ser Asp val ser Ala 805 810 815 lie lie Pro Tyr Ile Gly Pro Ala Leu Asn lie Ser Asn Ser val Arg 820 825 830 Arg Gly Asn Phe Thr Glu Ala Phe Ala val Thr Gly Val Thr lie Leu 835 840 845 Leu Glu Ala Phe Pro Glu Phe Thr ile Pro Ala Leu Gly Ala Phe val 850 855 860 Ile Tyr Ser Lys val Gln Glu Arg Asn Glu lie lie Lys Thr Ile Asp 865 870 875 880 Asn Cys Leu Glu Gln Arg lie Lys Arg Trp Lys Asp Ser Tyr Glu Trp 885 890 895 Met Met Gly Thr Trp Leu Ser Arg Ile ile Thr Gln Phe Asn Asn ile 900 905 910 Ser Tyr Gln Met Tyr Asp Ser Leu Asn Tyr Gln Ala Gly Ala Ile Lys 915 920 925 Ala Lys Ile Asp Leu Glu Tyr Lys Lys Tyr Ser Gly Ser Asp Lys Glu 930 935 940 Asn lie Lys Ser Gln val Glu Asn Leu Lys Asn Ser Leu Asp val Lys 945 950 955 960 lie Ser Glu Ala Met Asn Asn ile Asn Lys Phe ile Arg Glu Cys Ser 965 970 975 val Thr Tyr Leu Phe Lys Asn Met Leu Pro Lys Val lie Asp Glu Leu 980 985 990 Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu i lie : AST i Leu lie Asp 995 1000 1005

    Ser His Asn lie lie Leu Val 1015 Gly Glu Val Asp Lys 1020 Leu Lys Ala 1010 Lys val Asn Asn Ser Phe Gln Asn Thr lie Pro Phe Asn Ile Phe 1025 1030 1035 ser Tyr Thr Asn Asn ser Leu Leu Lys Asp Ile Ile Asn Glu Tyr 1040 1045 1050

    Page 95

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    68754 16 Dec 2015

    Phe Asn 1055 <210> 33 <211> 714 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic polypeptide <400> 33

    ΓΗ on r—Η

    Ο

    ΓΗ

    Glu Ser 1 Asn Gin Pro 5 Glu Lys Asn Gly Thr Ala Thr Lys Pro Glu Asn 10 15 ser Gly Asn Thr Thr Ser Glu Asn Gly Gin Thr Glu Pro Glu Lys Lys 20 25 30 Leu Glu Leu Arg Asn val ser Asp lie Glu Leu Tyr Ser Gin Thr Asn 35 40 45 Gly Thr Tyr Arg Gin His val Ser Leu Asp Gly lie pro Glu Asn Thr 50 55 60 Asp Thr Tyr Phe val Lys val Lys ser Ser Ala Phe Lys Asp Val Tyr 65 70 75 80 lie Pro val Ala Ser lie Thr Glu Glu Lys Arg Asn Gly Gin Ser val 85 90 95 Tyr Lys lie Thr Ala Lys Ala Glu Lys Leu Gin Gin Glu Leu Glu Asn 100 105 110 Lys Tyr val Asp Asn Phe Thr Phe Tyr Leu Asp Lys Lys Ala Lys Glu 115 120 125 Glu Asn Thr Asn Phe Thr ser Phe Ser Asn Leu val Lys Ala lie Asn 130 135 140 Gin Asn Pro Ser Gly Thr Tyr Hi s Leu Ala Ala Ser Leu Asn Ala Asn 145 150 155 160 Glu val Glu Leu Gly Pro Asp Glu Arg Ser Tyr lie Lys Asp Thr Phe 165 170 175 Thr Gly Arg Leu lie Gly Glu Lys Asp Gly Lys Asn Tyr Ala lie Tyr 180 185 190 Asn Leu Lys Lys Pro Leu phe Glu Asn Leu ser Gly Al a Thr Val Glu 195 200 205

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    Lys Leu Ser Leu Lys Asn val Ala Ile Ser Gly Lys Asn Asp lie Gly 210 215 220 Ser Leu Ala Asn Glu Ala Thr Asn Gly Thr Lys Ile Lys Gin Val Hi s 225 230 235 240 Val Asp Gly Cys val Asp Gly lie He Thr ser Lys Thr Lys Ser Asp 245 250 255 Asp Asp Asp Lys Asn Lys Ala Leu Asn Leu Gin cys Ile Lys Ile Lys 260 265 270 Asn Glu Asp Leu Thr Phe lie Ala Glu Lys Asn ser Phe Ser Glu Glu 275 280 285 Pro Phe Gin Asp Glu Ile Val Ser Tyr Asn Thr Lys Asn Lys Pro Leu 290 295 300 Asn phe Asn Tyr Ser Leu Asp Lys Ile lie val ASP Tyr Asn Leu Gin 305 310 315 320 Ser Lys Ile Thr Leu Pro Asn Asp Arg Thr Thr Pro val Thr Lys Gly 325 330 335 lie Pro Tyr Ala Pro Glu Tyr Lys Ser Asn Ala Ala ser Thr lie Glu 340 345 350 lie His Asn lie Asp Asp Asn Thr lie Tyr Gin Tyr Leu Tyr Ala Gin 355 360 365 Lys Ser Pro Thr Thr Leu Gin Arg Ile Thr Met Thr Asn Ser Val Asp 370 375 380 Asp Ala Leu Ile Asn Ser Thr Lys lie Tyr Ser Tyr Phe Pro Ser Val 385 390 395 400 lie Ser Lys val Asn Gin Gly Ala Gin Gly Ile Leu Phe Leu Gin Trp 405 410 415 val Arg Asp lie He Asp Asp Phe Thr Asn Glu Ser ser Gin Lys Thr 420 425 430 Thr lie Asp Lys Ile Ser Asp val Ser Thr lie Val pro Tyr lie Gly 435 440 445 Pro Ala Leu Asn lie Val Lys Gin Gly Tyr Glu Gly Asn Phe lie Gly 450 455 460 Ala Leu Glu Thr Thr Gly val val Leu Leu Leu Glu Tyr Ile Pro Glu 465 470 475 480

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    ile Thr Leu Pro val 485 lie Ala Ala Leu Ser Ile Ala Glu Ser ser Thr 490 495 Gin Lys Glu Lys ile lie Lys Thr lie Asp Asn Phe Leu Glu Lys Arg 500 505 510 Tyr Glu Lys Trp lie Glu val Tyr Lys Leu Val Lys Ala Lys Trp Leu 515 520 525 Gly Thr Val Asn Thr Gin Phe Gin Lys Arg Ser Tyr Gin Met Tyr Arg 530 535 540 Ser Leu Glu Tyr Gin val Asp Ala lie Lys Lys Ile Ile ASp Tyr Glu 545 550 555 560 Tyr Lys lie Tyr Ser Gly Pro Asp Lys Glu Gin ile Ala Asp Glu Ile 565 570 575 Asn Asn Leu Lys Asn Lys Leu Glu Glu Lys Ala Asn Lys Ala Met ile 580 585 590 Asn lie Asn Ile Phe Met Arg Glu Ser Ser Arg Ser Phe Leu Val Asn 595 600 605 Gin Met lie Asn Glu Ala Lys Lys Gin Leu Leu Glu Phe Asp Thr Gin 610 615 620 Ser Lys Asn lie Leu Met Gin Tyr lie Lys Ala Asn Ser Lys Phe He 625 630 635 640 Gly lie Thr Glu Leu Lys Lys Leu Glu Ser Lys lie Asn Lys val Phe 645 650 655 ser Thr Pro He Pro Phe Ser Tyr Ser Lys Asn Leu Asp cys Trp Val 660 665 670 Asp Asn Glu Glu Asp lie Asp val Leu Glu Gly Gly Gly Gly Ser Gly 675 680 685 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Val Ala Gly Cys Lys 690 695 700 Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 705 710

    <210> 34 <211> 710 <212> PRT <213> Artificial Sequence <220>

    <221> source

    Page 98

    2015268754 16 Dec 2015

    P31QO3au-pct.txt <223> /note=Description of Artificial sequence: synthetic polypeptide <400> 34

    Glu Ser Asn Gin Pro Glu Lys Asn Gly Thr Ala Thr Lys Pro Glu Asn 1 5 10 15 Ser Gly Asn Thr Thr Ser Glu Asn Gly Gin Thr Glu Pro Glu Lys Lys 20 25 30 Leu Glu Leu Arg Asn Val Ser Asp lie Glu Leu Tyr Ser Gin Thr Asn 35 40 45 Gly Thr Tyr Arg Gin His Val Ser Leu Asp Gly He Pro Glu Asn Thr 50 55 60 Asp Thr Tyr Phe val Lys Val Lys Ser Ser Ala Phe Lys Asp val Tyr 65 70 75 80 lie Pro val Ala Ser lie Thr Glu Glu Lys Arg Asn Gly Gin Ser Val 85 90 95 Tyr Lys lie Thr Ala Lys Ala Glu Lys Leu Gin Gin Glu Leu Glu Asn 100 105 110 Lys Tyr val ASp Asn Phe Thr Phe Tyr Leu Asp Lys Lys Ala Lys Glu 115 120 125 Glu Asn Thr Asn Phe Thr Ser Phe Ser Asn Leu val Lys Ala lie Asn 130 135 140 Gin Asn Pro ser Gly Thr Tyr Hi s Leu Ala Ala ser Leu Asn Ala Asn 145 150 155 160 Glu val Glu Leu Gly Pro Asp Glu Arg ser Tyr lie Lys Asp Thr Phe 165 170 175 Thr Gly Arg Leu lie Gly Glu Lys Asp Gly Lys Asn Tyr Ala lie Tyr 180 185 190 Asn Leu Lys Lys Pro Leu Phe Glu Asn Leu Ser Gly Ala Thr val Glu 195 200 205 Lys Leu Ser Leu Lys Asn Val Ala lie Ser Gly Lys Asn Asp lie Gly 210 215 220 ser Leu Ala Asn Glu Ala Thr Asn Gly Thr Lys lie Lys Gin val Hi s 225 230 235 240 val Asp Gly cys Val Asp Gly lie lie Thr ser Lys Thr Lys Ser Asp 245 250 255

    Page 99

    ASp Asp Asp Lys 260 Ala Gly Cys Lys P31OO3au-pct.txt Asn phe Phe Trp Lys 265 Thr 270 Phe Thr Ser cys Ala Leu Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 275 280 285 Gly Gly Gly Ser Ala Leu Ala Leu Gin Cys lie Lys lie Lys Asn Glu 290 295 300 Asp Leu Thr Phe lie Ala Glu Lys Asn Ser Phe Ser Glu Glu pro Phe 305 310 315 320 Gin Asp Glu He val Ser Tyr Asn Thr Lys Asn Lys Pro Leu Asn Phe 325 330 335 Asn Tyr Ser Leu Asp Lys lie He Val Asp Tyr Asn Leu Gin Ser Lys 340 345 350 lie Thr Leu Pro Asn Asp Arg Thr Thr Pro val Thr Lys Gly lie Pro 355 360 365 Tyr Ala pro Gl U Tyr Lys Ser Asn Ala Ala Ser Thr lie Glu lie Hi s 370 375 380 Asn lie Asp Asp Asn Thr lie Tyr Gin Tyr Leu Tyr Ala Gin Lys Ser 385 390 395 400 Pro Thr Thr Leu Gin Arg lie Thr Met Thr Asn Ser val Asp Asp Ala 405 410 415 Leu lie Asn Ser Thr Lys lie Tyr Ser Tyr Phe Pro ser Val He Ser 420 425 430 Lys Val Asn Gin Gly Ala Gin Gly lie Leu Phe Leu Gin Trp Val Arg 435 440 445 Asp lie lie Asp Asp Phe Thr Asn Glu Ser ser Gin Lys Thr Thr He 450 455 460 Asp Lys lie Ser Asp val ser Thr lie val Pro Tyr lie Gly Pro Ala 465 470 475 480 Leu Asn lie val Lys Gin Gly Tyr Glu Gly Asn Phe lie Gly Ala Leu 485 490 495 Glu Thr Thr Gly Val Val Leu Leu Leu Glu Tyr lie Pro Glu lie Thr 500 505 510 Leu Pro Val He Ala Ala Leu Ser lie Ala Glu Ser ser Thr Gin Lys 515 520 525

    Page 100

    2015268754 16 Dec 2015

    Glu Lys Ile P31OO3au-pct.txt lie Lys Thr lie Asp Asn 535 Phe Leu Glu 540 Lys Arg Tyr Glu 530 Lys Trp ile Glu val Tyr Lys Leu val Lys Ala Lys Trp Leu Gly Thr 545 550 555 560 val Asn Thr Gin Phe Gin Lys Arg Ser Tyr Gin Met Tyr Arg Ser Leu 565 570 575 Glu Tyr Gin Val Asp Ala lie Lys Lys lie ile Asp Tyr Glu Tyr Lys 580 585 590 He Tyr Ser Gly Pro Asp Lys Glu Gin ile Ala Asp Glu Ile Asn Asn 595 600 605 Leu Lys Asn Lys Leu Glu Glu Lys Ala Asn Lys Ala Met Ile Asn lie 610 615 620 Asn Ile Phe Met Arg Glu Ser Ser Arg Ser Phe Leu val Asn Gin Met 625 630 635 640 Ile Asn Glu Ala Lys Lys Gin Leu Leu Glu Phe Asp Thr Gin Ser Lys 645 650 655 Asn lie Leu Met Gin Tyr lie Lys Ala Asn Ser Lys Phe lie Gly lie 660 665 670 Thr Glu Leu Lys Lys Leu Glu Ser Lys lie Asn Lys val Phe Ser Thr 675 680 685 Pro Ile Pro Phe Ser Tyr Ser Lys Asn Leu Asp Cys Trp Val Asp Asn 690 695 700 Glu Glu Asp lie Asp Val 705 710

    <210> 35 <211> 898 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polypeptide <400> 35

    Glu Phe Val Asn 1 Lys Gin 5 Phe Asn Tyr Lys Asp 10 Pro Val Asn Gly 15 val Asp lie Ala Tyr Ile Lys lie Pro Asn Ala Gly Gin Met Gin Pro Val 20 25 30 Lys Ala Phe Lys Ile Hi s Asn Lys lie Trp val lie Pro Glu Arg Asp

    Page 101

    P31OO3au-pct.txt

    35 40 45

    Ο

    Thr Phe Thr Asn Pro val Glu Ser 70 Glu Gly Asp Leu Asn 55 pro Pro Pro Glu Ala 60 O <D 50 Tyr Tyr Asp ser Thr 75 Gin Val Pro Tyr Leu Ser Thr Asp 80 Asn τι- Glu Lys Asp Asn 85 Tyr Leu Lys Gly Val 90 Thr Lys Leu Phe Glu 95 Arg 00 ^l1e kO Tyr ser Thr 100 Asp Leu Gly Arg Met Leu 105 Leu Thr Ser lie 110 Val Arg CH Ω Gly O Ile Pro 115 Phe Trp Gly Gly Ser Thr lie 120 Asp Thr Glu 125 Leu Lys Val CH lie Asp 130 Thr Asn cys lie Asn 135 val Ile Gin Pro Asp 140 Gly Ser Tyr Arg Ser 145 Glu Glu Leu Asn Leu 150 val ile ile Gly Pro 155 Ser Ala Asp lie Ile 160 Gin Phe Glu cys Leu 165 Ser Phe Gly His Glu 170 Val Leu Asn Leu Thr 175 Arg Asn Gly Tyr Gly 180 Ser Thr Gin Tyr lie Arg 185 Phe Ser Pro Asp 190 Phe Thr Phe Gly Phe 195 Glu Glu ser Leu Glu Val Asp 200 Thr Asn Pro 205 Leu Leu Gly Ala Gly 210 Lys Phe Ala Thr Asp 215 Pro Ala val Thr Leu 220 Ala Hi s Glu Leu lie 225 His Ala Gly Hi s Arg 230 Leu Tyr Gly lie Ala 235 Ile Asn Pro Asn Arg 240 val Phe Lys val Asn 245 Thr Asn Ala Tyr Tyr 250 Glu Met Ser Gly Leu 255 Glu val Ser Phe Glu 260 Glu Leu Arg Thr Phe Gly 265 Gly Hi s Asp Ala 270 Lys Phe lie Asp ser 275 Leu Gin Glu Asn Glu Phe Arg 280 Leu Tyr Tyr 285 Tyr Asn Lys Phe Lys 290 ASp Ile Ala Ser Thr 295 Leu Asn Lys Ala Lys 300 Ser lie Val Gly Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn val Phe Lys Glu Lys Tyr

    Page 102

    2015268754 16 Dec 2015

    305 P31OO3au-pct.txt 310 315 320 Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu lie Tyr Thr Glu Asp Asn 340 345 350 Phe val Lys Phe Phe Lys val Leu Asn Arg Lys Thr Tyr Leu Asn Phe 355 360 365 Asp Lys Ala Val Phe Lys Ile Asn Ile val Pro Lys Val Asn Tyr Thr 370 375 380 Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe 385 390 395 400 Asn Gly Gln Asn Thr Glu Ile Asn Asn Met Asn Phe Thr Lys Leu Lys 405 410 415 Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys Val Asp Gly 420 425 430 He Ile Thr Ser Lys Thr Lys ser ASp Asp Asp Asp Lys Asn Lys Ala 435 440 445 Leu Asn Leu Gln cys lie Lys Val Asn Asn Trp Asp Leu phe Phe ser 450 455 460 Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu lie 465 470 475 480 Thr Ser Asp Thr Asn lie Glu Ala Ala Glu Glu Asn lie Ser Leu Asp 485 490 495 Leu Ile Gln Gln Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro Glu 500 505 510 Asn lie Ser Ile Glu Asn Leu Ser Ser Asp Ile Ile Gly Gln Leu Glu 515 520 525 Leu Met Pro Asn lie Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu Leu 530 535 540 Asp Lys Tyr Thr Met Phe Hi s Tyr Leu Arg Ala Gln Glu phe Glu His 545 550 555 560 Gly Lys Ser Arg lie Ala Leu Thr Asn Ser val Asn Glu Ala Leu Leu 565 570 575 Asn Pro Ser Arg Val Tyr Thr Phe Phe Ser ser Asp Tyr val Lys Lys

    Page 103

    P31003au-pct.txt

    IT)

    Ο (Μ ο

    <υ

    Ω ήΙΤ)

    1^ οο kO (Μ

    ΙΤ)

    Ο (Μ

    580 585 590 val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu Gln 595 600 605 Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr Asp 610 615 620 Lys Ile Ala Asp lie Thr Ile lie lie Pro Tyr Ile Gly Pro Ala Leu 625 630 635 640 Asn Ile Gly Asn Met Leu Tyr Lys Asp Asp Phe Val Gly Ala Leu Ile 645 650 655 Phe Ser Gly Ala Val ile Leu Leu Glu Phe Ile Pro Glu lie Ala Ile 660 665 670 Pro Val Leu Gly Thr Phe Ala Leu Val ser Tyr lie Ala Asn Lys Val 675 680 685 Leu Thr Val Gln Thr lie Asp Asn Ala Leu Ser Lys Arg Asn Glu Lys 690 695 700 Trp Asp Glu Val Tyr Lys Tyr lie val Thr Asn Trp Leu Ala Lys val 705 710 715 720 Asn Thr Gln lie Asp Leu Ile Arg Lys Lys Met Lys Glu Ala Leu Glu 725 730 735 Asn Gln Ala Glu Ala Thr Lys Ala Ile Ile Asn Tyr Gln Tyr Asn Gln 740 745 750 Tyr Thr Glu Glu Glu Lys Asn Asn Ile Asn Phe Asn lie Asp Asp Leu 755 760 765 Ser ser Lys Leu Asn Glu Ser lie Asn Lys Ala Met lie Asn lie Asn 770 775 780 Lys Phe Leu Asn Gln Cys Ser Val Ser Tyr Leu Met Asn Ser Met lie 785 790 795 800 pro Tyr Gly Val Lys Arg Leu Glu Asp Phe Asp Ala Ser Leu Lys Asp 805 810 815 Ala Leu Leu Lys Tyr ile Tyr Asp Asn Arg Gly Thr Leu lie Gly Gln 820 825 830 val Asp Arg Leu Lys Asp Lys val Asn Asn Thr Leu Ser Thr Asp ile 835 840 845 pro Phe Gln Leu Ser Lys Tyr val Asp Asn Gln Arg Leu Leu Ser Thr

    Page 104

    2015268754 16 Dec 2015

    850

    855

    P31OO3au-pct.txt

    860

    Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly ser Gly Gly Gly Gly 865 870 875 880

    Ser Ala Leu Val Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr 885 890 895

    Ser Cys <210> 36 <211> 943 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 36

    Glu 1 Phe Val Asn Lys Gin Phe Asn Tyr Lys Asp Pro Val Asn Gly 15 Val 5 10 ASp Ile Ala Tyr Ile Lys lie Pro Asn Ala Gly Gin Met Gin Pro Val 20 25 30 Lys Ala Phe Lys lie Hi s Asn Lys lie Trp val lie Pro Glu Arg Asp 35 40 45 Thr Phe Thr Asn Pro Glu Glu Gly ASp Leu Asn Pro Pro Pro Glu Ala 50 55 60 Lys Gin val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr Asp 65 70 75 80 Asn Glu Lys Asp Asn Tyr Leu Lys Gly val Thr Lys Leu Phe Glu Arg 85 90 95 lie Tyr Ser Thr ASp Leu Gly Arg Met Leu Leu Thr Ser lie val Arg 100 105 110 Gly lie Pro Phe Trp Gly Gly ser Thr lie Asp Thr Glu Leu Lys val 115 120 125 lie Asp Thr Asn cys Ile Asn Val lie Gin Pro Asp Gly Ser Tyr Arg 130 135 140 Ser Glu Glu Leu Asn Leu val Ile lie Gly Pro ser Ala Asp lie Ile 145 150 155 160 Gin Phe Glu Cys Leu Ser Phe Gly Hi s Glu Val Leu Asn Leu Thr Arg

    165 170 175

    Page 105

    IT)

    P31003au-pct.txt

    o Asn Gly Tyr Gly Ser Thr Gin Tyr Ile Arg Phe Ser Pro Asp Phe Thr Cn 180 185 190 o CD n Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro Leu Leu Gly 195 200 205 Ala Gly Lys Phe Ala Thr Asp Pro Ala val Thr Leu Ala Hi s Glu Leu 210 215 220 τ|- ΐ/Ί ile His Ala Gly Hi s Arg Leu Tyr Gly Ile Ala lie Asn Pro Asn Arg 00 225 230 235 240 <o C4 K'l val Phe Lys val Asn Thr Asn Ala Tyr Tyr Glu Met ser Gly Leu Glu ” i 245 250 255 o C4 Val Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly Hi s Asp Ala Lys Phe 260 265 270 lie Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn Lys 275 280 285 Phe Lys Asp lie Ala Ser Thr Leu Asn Lys Ala Lys Ser lie Val Gly 290 295 300 Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn val Phe Lys Glu Lys Tyr 305 310 315 320 Leu Leu ser Glu Asp Thr Ser Gly Lys Phe ser val Asp Lys Leu Lys 325 330 335 Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu Ile Tyr Thr Glu Asp Asn 340 345 350 Phe val Lys Phe Phe Lys Val Leu Asn Arg Lys Thr Tyr Leu Asn Phe 355 360 365 Asp Lys Ala Val Phe Lys ile Asn ile val Pro Lys Val Asn Tyr Thr 370 375 380 Ile Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala Ala Asn Phe 385 390 395 400 Asn Gly Gin Asn Thr Glu ile Asn Asn Met Asn phe Thr Lys Leu Lys 405 410 415 Asn phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu cys Val Asp Gly 420 425 430 lie lie Thr Ser Lys Thr Lys Ser Asp Asp Asp Asp Lys Asn Lys Ala 435 440 445

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    Leu Asn Leu 450 Gin Cys lie Lys 455 val Asn Asn Trp Asp 460 Leu Phe Phe ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly Glu Glu lie 465 470 475 480 Thr Ser Asp Thr Asn lie Glu Ala Ala Glu Glu Asn lie ser Leu Asp 485 490 495 Leu lie Gin Gin Tyr Tyr Leu Thr Phe Asn Phe Asp Asn Glu Pro Glu 500 505 510 Asn lie Ser lie Glu Asn Leu Ser Ser Asp lie He Gly Gin Leu Glu 515 520 525 Leu Met Pro Asn He Glu Arg Phe Pro Asn Gly Lys Lys Tyr Glu Leu 530 535 540 Asp Lys Tyr Thr Met Phe Hi s Tyr Leu Arg Ala Gin Glu Phe Glu Hi s 545 550 555 560 Gly Lys Ser Arg lie Ala Leu Thr Asn ser Val Asn Glu Ala Leu Leu 565 570 575 Asn Pro Ser Arg val Tyr Thr Phe Phe ser ser Asp Tyr val Lys Lys 580 585 590 Val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu Gly Trp Val Glu Gin 595 600 605 Leu val Tyr Asp Phe Thr Asp Glu Thr Ser Glu Val Ser Thr Thr Asp 610 615 620 Lys lie Ala Asp lie Thr He lie lie Pro Tyr lie Gly Pro Ala Leu 625 630 635 640 Asn lie Gly Asn Met Leu Tyr Lys Asp Asp Phe val Gly Ala Leu lie 645 650 655 Phe ser Gly Ala Val lie Leu Leu Glu Phe lie Pro Glu lie Ala lie 660 665 670 Pro val Leu Gly Thr Phe Ala Leu val Ser Tyr lie Ala Asn Lys val 675 680 685 Leu Thr Val Gin Thr He Asp Asn Ala Leu ser Lys Arg Asn Glu Lys 690 695 700 Trp Asp Glu Val Tyr Lys Tyr lie Val Thr Asn Trp Leu Ala Lys val 705 710 715 720

    Page 107

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    Asn Thr Gin lie Asp Leu lie 725 Arg Lys Lys Met Lys Glu Ala Leu Glu 730 735 Asn Gin Ala Glu Ala Thr Lys Ala lie lie Asn Tyr Gin Tyr Asn Gin 740 745 750 Tyr Thr Glu Glu Glu Lys Asn Asn lie Asn Phe Asn He ASP Asp Leu 755 760 765 ser Ser Lys Leu Asn Glu Ser lie Asn Lys Ala Met lie Asn lie Asn 770 775 780 Lys Phe Leu Asn Gin Cys Ser val Ser Tyr Leu Met Asn Ser Met lie 785 790 795 800 Pro Tyr Gly Val Lys 805 Arg Leu Glu Asp Phe 810 Asp Ala Ser Leu Lys 815 Asp Al a Leu Leu Lys Tyr lie Tyr Asp Asn Arg Gly Thr Leu lie Gly Gin 820 825 830 Val Asp Arg Leu Lys Asp Lys Val Asn Asn Thr Leu Ser Thr Asp lie 835 840 845 Pro Phe Gin Leu Ser Lys Tyr Val Asp Asn Gin Arg Leu Leu Ser Thr 850 855 860 Leu Glu Gly Gly Gly Gly ser Gly Gly Gly Gly Ser Gly Gly Gly Gl y 865 870 875 880 Ser Gly Gly Gly Gly ser Ala Leu Val Asp Asn Ser Asp pro Lys Cys 885 890 895 Pro Leu ser His Glu Gly Tyr Cys Leu Asn Asp Gly Val cys Met Tyr 900 905 910 He Gly Thr Leu Asp Arg Tyr Ala Cys Asn cys val val Gly Tyr Val 915 920 925 Gly Glu Arg Cys Gin Tyr Arg Asp Leu Lys Leu Ala Glu Leu Arg 930 935 940

    <210> 37 <211> 1061 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polypeptide

    Page 108

    P31OO3au-pct.txt <400> 37

    2015268754 16 Dec 2015

    Pro Lys Ile Asn Ser Phe Asn Tyr Asn Asp Pro val Asn Asp Arg Thr 1 5 10 15 Ile Leu Tyr Ile Lys Pro Gly Gly Cys Gin Glu Phe Tyr Lys Ser Phe 20 25 30 Asn lie Met Lys Asn Ile Trp lie lie Pro Glu Arg Asn Val lie Gly 35 40 45 Thr Thr Pro Gl n Asp Phe Hi s Pro Pro Thr Ser Leu Lys Asn Gly Asp 50 55 60 ser Ser Tyr Tyr Asp Pro Asn Tyr Leu Gin Ser ASp Glu Glu Lys Asp 65 70 75 80 Arg Phe Leu Lys Ile val Thr Lys lie Phe Asn Arg Ile Asn Asn Asn 85 90 95 Leu ser Gly Gly Ile Leu Leu Glu Glu Leu Ser Lys Ala Asn Pro Tyr 100 105 110 Leu Gly Asn ASp Asn Thr Pro Asp Asn Gin Phe Hi s Ile Gly Asp Ala 115 120 125 ser Al a Val Glu lie Lys Phe Ser Asn Gly ser Gin Hi s lie Leu Leu 130 135 140 Pro Asn val lie lie Met Gly Ala Glu Pro Asp Leu Phe Glu Thr Asn 145 150 155 160 ser ser Asn lie Ser Leu Arg Asn Asn Tyr Met Pro Ser Asn His Gly 165 170 175 Phe Gly Ser lie Ala Ile Val Thr Phe Ser Pro Glu Tyr Ser Phe Arg 180 185 190 Phe Asn Asp Asn Ser Ile Asn Glu Phe Ile Gin Asp Pro Ala Leu Thr 195 200 205 Leu Met His Glu Leu lie Hi s ser Leu His Gly Leu Tyr Gly Ala Lys 210 215 220 Gly Ile Thr Thr Thr cys Ile Ile Thr Gin Gin Gin Asn Pro Leu Ile 225 230 235 240 Thr Asn Arg Lys Gly Ile Asn Ile Glu Glu Phe Leu Thr Phe Gly Gly 245 250 255 Asn Asp Leu Asn lie Ile Thr Val Ala Gin Tyr Asn Asp He Tyr Thr 260 265 270

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    o ^Asn CM o Leu val 290 Leu Asn Asp Tyr Arg Lys Ile Ala Ser Lys Leu Ser Lys val 275 ser Asn Pro 280 Asp 300 285 A Gln Gln Leu 295 Asn Pro Tyr Lys Ile Phe Gln Glu Lys 305 Tyr Gly Leu Asp Lys 310 Asp Ala ser Gly lie 315 Tyr Ser val Asn Ile 320 ’rf- Asn Lys Phe Asp Asp 325 Ile Leu Lys Lys Leu 330 Tyr ser Phe Thr Glu 335 Phe oo CM IT) Asp Leu Al a Thr 340 Lys Phe Gln val Lys Cys 345 Arg Glu Thr Tyr 350 ile Gly CM Gln Tyr Lys 355 Tyr Phe Lys Leu Ser 360 Asn Leu Leu Asn Asp 365 Ser lie Tyr Asn Ile 370 Ser Glu Gly Tyr Asn 375 Ile Asn Asn Leu Lys 380 val Asn Phe Arg Gly 385 Gln Asn Ala Asn Leu 390 Asn Pro Arg ile lie 395 Lys Pro lie Thr Gly 400 Arg Gly Leu Val Lys 405 Lys Ile Ile Arg Phe 410 Cys val Asp Gly lie 415 Ile Thr Ser Lys Thr 420 Lys ser Leu ile Glu Gly 425 Arg Asn Lys Al a 430 Leu Asn Leu Gln cys 435 Ile Glu lie Asn Asn 440 Gly Glu Leu Phe Phe 445 Val Ala Ser Glu Asn 450 ser Tyr Asn Asp Asp 455 Asn Ile Asn Thr Pro 460 Lys Glu lie Asp Asp 465 Thr val Thr ser Asn 470 Asn Asn Tyr Glu Asn 475 Asp Leu Asp Gln val 480 He Leu Asn Phe Asn 485 Ser Glu Ser Ala Pro 490 Gly Leu ser Asp Glu 495 Lys Leu Asn Leu Thr 500 lie Gln Asn Asp Ala Tyr 505 lie Pro Lys Tyr 510 Asp Ser Asn Gly Thr 515 ser Asp lie Glu Gln 520 His Asp Val Asn Glu 525 Leu Asn val Phe Phe 530 Tyr Leu Asp Ala Gln 535 Lys Val Pro Glu Gly 540 Glu Asn Asn val

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    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Asn Leu Thr ser Ser Ile Asp Thr Ala Leu Leu Gl u Gln Pro Lys Ile 545 550 555 560 Tyr Thr Phe Phe Ser Ser Glu Phe lie Asn Asn Val Asn Lys Pro val 565 570 575 Gln Ala Ala Leu Phe Val Ser Trp lie Gln Gln val Leu val Asp Phe 580 585 590 Thr Thr Glu Ala Asn Gln Lys Ser Thr val Asp Lys Ile Ala Asp lie 595 600 605 Ser Ile val val Pro Tyr Ile Gly Leu Ala Leu Asn Ile Gly Asn Glu 610 615 620 Ala Gln Lys Gly Asn Phe Lys Asp Ala Leu Glu Leu Leu Gly Ala Gly 625 630 635 640 ile Leu Leu Glu Phe Glu Pro Glu Leu Leu Ile Pro Thr Ile Leu Val 645 650 655 Phe Thr lie Lys ser Phe Leu Gly Ser Ser Asp Asn Lys Asn Lys val 660 665 670 lie Lys Ala lie Asn Asn Ala Leu Lys Glu Arg Asp Glu Lys Trp Lys 675 680 685 Glu val Tyr Ser Phe Ile val ser Asn Trp Met Thr Lys lie Asn Thr 690 695 700 Gln Phe Asn Lys Arg Lys Glu Gln Met Tyr Gln Ala Leu Gln Asn Gln 705 710 715 720 Val Asn Ala lie Lys Thr lie Ile Glu Ser Lys Tyr Asn Ser Tyr Thr 725 730 735 Leu Glu Glu Lys Asn Glu Leu Thr Asn Lys Tyr Asp lie Lys Gln lie 740 745 750 Glu Asn Glu Leu Asn Gln Lys val Ser Ile Ala Met Asn Asn Ile Asp 755 760 765 Arg Phe Leu Thr Glu Ser Ser Ile Ser Tyr Leu Met Lys lie lie Asn 770 775 780 Glu val Lys lie Asn Lys Leu Arg Glu Tyr Asp Glu Asn val Lys Thr 785 790 795 800 Tyr Leu Leu Asn Tyr Ile ile Gln His Gly Ser Ile Leu Gly Glu Ser

    805 810 815

    Page 111

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Gin Gin Glu Leu 820 Asn Ser Met Val Pro Phe Lys 835 Leu Ser ser Tyr Thr 840 Phe Asn 850 Lys Phe phe Lys Leu 855 Glu Gly 865 Ser Gly Gly Gly Gly 870 Ser Gly Pro Gly Glu Asp Ser 885 Lys Asp Val Thr Ser Ser Glu 900 Arg ile Asp Lys lie Ser Ala 915 Leu Arg Lys Glu Thr 920 Ser Ser 930 Lys Glu Al a Leu Ala 935 Glu Al a 945 Glu Lys ASP Gly Cys 950 Phe Gin Leu Val Lys lie lie 965 Thr Gly Leu Tyr Leu Gin Asn 980 Arg Phe Glu ser Gin Met Ser 995 Thr Lys val Leu Ile 100)

    Thr 825 Asp Thr Leu Asn Asn 830 ser lie Asp Asp Lys Ile Leu 845 ile Ser Tyr Gly Gly Gly Gly 860 ser Gly Gly Gly Gly Gly Gly 875 Ser Ala Leu val Pro 880 Ala Ala 890 Pro Hi s Arg Gin Pro 895 Leu Gin 905 lie Arg Tyr lie Leu 910 Asp Gly Cys Asn Lys Ser Asn 925 Met cys Glu Asn Asn Leu Asn 940 Leu Pro Lys Met Ser Gly Phe 955 Asn Glu Glu Thr cys 960 Leu Glu 970 Phe Glu val Tyr Leu 975 Glu ser Glu Gl u Gin Ala Arg Ala Val

    985 990

    Gin Phe Leu Gin Lys Lys Ala Lys 1005

    Asn Leu Asp Ala He Thr Thr Pro Asp Pro Thr Thr Asn Ala Ser 1010 1015 1020 Leu Leu Thr Lys Leu Gin Ala Gin Asn Gin Trp Leu Gin Asp Met 1025 1030 1035 Thr Thr Hi s Leu lie Leu Arg Ser Phe Lys Glu Phe Leu Gin ser 1040 1045 1050 Ser Leu Arg Ala Leu Arg Gin Met 1055 1060

    <210> 38 <211> 963 <212> PRT

    Page 112

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <213> Artificial Sequence <22O>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polypeptide <400> 38

    Pro val Thr lie Asn Asn Phe Asn Tyr Asn Asp Pro lie Asp Asn Asn 15 10 15

    Asn lie lie Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Arg Tyr 20 25 30

    Tyr Lys Ala Phe Lys lie Thr Asp Arg lie Trp Ile Ile Pro Glu Arg 35 40 45

    Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly lie 50 55 60

    Phe Asn Arg Asp val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn Thr

    65 70 75 80

    Asn Asp Lys Lys Asn lie Phe Leu Gin Thr Met lie Lys Leu Phe Asn

    85 90 95

    Arg ile Lys ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met Ile Ile 100 105 110

    Asn Gly lie Pro Tyr Leu Gly Asp Arg Arg val Pro Leu Glu Glu Phe 115 120 125

    Asn Thr Asn Ile Ala Ser val Thr Val Asn Lys Leu Ile Ser Asn Pro 130 135 140

    Gly Glu val Glu Arg Lys Lys Gly lie Phe Ala Asn Leu Ile Ile Phe

    145 150 155 160

    Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr Ile Asp Ile Gly lie

    165 170 175

    Gin Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly lie Met Gin Met 180 185 190

    Lys Phe Cys Pro Glu Tyr val ser val Phe Asn Asn Val Gin Glu Asn 195 200 205

    Lys Gly Ala Ser Ile Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro Ala 210 215 220

    Leu lie Leu Met His Glu Leu lie His val Leu His Gly Leu Tyr Gly 225 230 235 240

    Page 113

    lz*\ P3H 3O3ai j-pc t .txt i lie Lys val Asp Asp Leu Pro lie val Pro Asn Glu Lys Lys Phe Phe o 245 250 255 CM Q Met Gin ser Thr Asp Ala lie Gin Ala Glu Glu Leu Tyr Thr Phe Gly <L> Q 260 265 270 Gly Gin Asp Pro Ser lie lie Thr Pro ser Thr Asp Lys Ser lie Tyr ,-H 275 280 285 Tj- Asp Lys val Leu Gin Asn phe Arg Gly lie Val Asp Arg Leu Asn Lys >n 290 295 300 oo val Leu val cys lie Ser ASp Pro Asn lie Asn lie Asn lie Tyr Lys CM 305 310 315 320 in o Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly Lys CM 325 330 335 Tyr Ser lie ASp val Glu ser Phe Asp Lys Leu Tyr Lys ser Leu Met 340 345 350 Phe Gly Phe Thr Glu Thr Asn lie Ala Glu Asn Tyr Lys lie Lys Thr 355 360 365 Arg Ala Ser Tyr Phe Ser Asp ser Leu Pro Pro val Lys lie Lys Asn 370 375 380 Leu Leu Asp Asn Glu lie Tyr Thr lie Glu Glu Gly Phe Asn lie ser 385 390 395 400 Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gin Asn Lys Ala lie Asn 405 410 415 Lys Gin Ala Tyr Glu Glu lie Ser Lys Glu His Leu Al a val Tyr Lys 420 425 430 lie Gl n Met cys val Asp Gly lie lie Thr ser Lys Thr Lys Ser Leu 435 440 445 lie Glu Gly Arg Asn Lys Ala Leu Asn Leu Gin cys lie Asp Val Asp 450 455 460 Asn Glu Asp Leu Phe Phe lie Ala Asp Lys Asn Ser Phe Ser Asp Asp 465 470 475 480 Leu Ser Lys Asn Glu Arg lie Glu Tyr Asn Thr Gin Ser Asn Tyr lie 485 490 495 Glu Asn Asp Phe Pro lie Asn Glu Leu lie Leu Asp Thr Asp Leu lie 500 505 510

    Page 114

    2015268754 16 Dec 2015

    Ser Lys lie 515 P31OO3au-pct.txt Glu Leu Pro Ser Glu Asn Thr Glu Ser Leu Thr Asp Phe 520 525 Asn Val Asp Val Pro val Tyr Glu Lys Gin Pro Ala lie Lys Lys lie 530 535 540 Phe Thr ASp Glu Asn Thr lie Phe Gin Tyr Leu Tyr Ser Gin Thr Phe 545 550 555 560 Pro Leu Asp lie Arg Asp lie Ser Leu Thr Ser Ser Phe Asp Asp Ala 565 570 575 Leu Leu Phe Ser Asn Lys Val Tyr Ser Phe Phe Ser Met Asp Tyr He 580 585 590 Lys Thr Ala Asn Lys val val Glu Ala Gly Leu Phe Ala Gly Trp val 595 600 605 Lys Gin lie Val Asn Asp Phe Val lie Glu Ala Asn Lys Ser Asn Thr 610 615 620 Met Asp Lys lie Ala Asp lie Ser Leu lie val Pro Tyr lie Gly Leu 625 630 635 640 Ala Leu Asn val Gly Asn Glu Thr Ala Lys Gly Asn Phe Glu Asn Ala 645 650 655 Phe Glu lie Ala Gly Ala Ser He Leu Leu Glu Phe lie Pro Glu Leu 660 665 670 Leu lie Pro Val Val Gly Ala Phe Leu Leu Glu Ser Tyr lie Asp Asn 675 680 685 Lys Asn Lys lie lie Lys Thr lie Asp Asn Ala Leu Thr Lys Arg Asn 690 695 700 Glu Lys Trp Ser Asp Met Tyr Gly Leu lie Val Ala Gin Trp Leu Ser 705 710 715 720 Thr val Asn Thr Gin Phe Tyr Thr lie Lys Glu Gly Met Tyr Lys Ala 725 730 735 Leu Asn Tyr Gin Ala Gin Ala Leu Glu Glu lie He Lys Tyr Arg Tyr 740 745 750 Asn lie Tyr Ser Glu Lys Glu Lys Ser Asn lie Asn lie Asp Phe Asn 755 760 765 Asp lie Asn Ser Lys Leu Asn Glu Gly lie Asn Gin Ala lie Asp Asn 770 775 780

    Page 115

    2015268754 16 Dec 2015

    P31I D03ai j-pc t.txt He Asn Asn Phe lie Asn Gly Cys Ser Val Ser Tyr Leu Met Lys Lys 785 790 795 800 viet ile Pro Leu Al a Val Glu Lys Leu Leu Asp Phe Asp Asn Thr Leu 805 810 815 Lys Lys Asn Leu Leu Asn Tyr lie Asp Glu Asn Lys Leu Tyr Leu lie 820 825 830 Gly Ser Ala Glu Tyr Glu Lys Ser Lys Val Asn Lys Tyr Leu Lys Thr 835 840 845 ile Met Pro Phe Asp Leu ser lie Tyr Thr Asn Asp Thr Ile Leu lie 850 855 860 Glu Met Phe Asn Lys Tyr Asn ser Leu Glu Gly Gly Gly Gly Ser Gly 865 870 875 880 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Val Ala Lys Glu Leu 885 890 895 Arg Cys Gln Cys 900 Ile Lys Thr Tyr Ser 905 Lys Pro Phe Hi s pro 910 Lys Phe lie Lys Glu Leu Arg Val lie Glu Ser Gly Pro Hi s cys Ala Asn Thr 915 920 925 Glu Ile lie val Lys Leu Ser Asp Gly Arg Glu Leu cys Leu Asp Pro 930 935 940 Lys Glu Asn Trp val Gln Arg Val Val Glu Lys Phe Leu Lys Arg Ala 945 950 955 960

    Glu Asn Ser

    J <210> 39 <211> 959 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial sequence: Synthetic polypeptide <400> 39

    Pro val Ala lie Asn Ser Phe Asn Tyr Asn Asp Pro 10 Val Asn Asp 15 Asp 1 5 Thr lie Leu Tyr Met Gln Ile Pro Tyr Glu Glu Lys Ser Lys Lys Tyr 20 25 30 Tyr Lys Ala Phe Glu lie Met Arg Asn Val Trp lie Ile pro Glu Arg

    Page 116

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    35 40 45

    Asn Thr 50 lie Gly Thr Asn Pro Ser Asp 55 Phe Asp Pro Pro Ala 60 Ser Leu Lys Asn Gly Ser ser Ala Tyr Tyr Asp Pro Asn Tyr Leu Thr Thr Asp 65 70 75 80 Ala Glu Lys Asp Arg Tyr Leu Lys Thr Thr lie Lys Leu Phe Lys Arg 85 90 95 lie Asn Ser Asn Pro Ala Gly Lys val Leu Leu Gin Glu lie Ser Tyr 100 105 110 Ala Lys Pro Tyr Leu Gly Asn Asp Hi s Thr Pro lie Asp Glu Phe Ser 115 120 125 Pro Val Thr Arg Thr Thr Ser val Asn lie Lys Leu Ser Thr Asn Val 130 135 140 Glu Ser Ser Met Leu Leu Asn Leu Leu val Leu Gly Ala Gly Pro Asp 145 150 155 160 lie Phe Glu Ser Cys Cys Tyr Pro val Arg Lys Leu lie Asp Pro ASp 165 170 175 val val Tyr Asp Pro Ser Asn Tyr Gly Phe Gly ser lie Asn lie val 180 185 190 Thr Phe Ser Pro Glu Tyr Glu Tyr Thr Phe Asn Asp lie Ser Gly Gly 195 200 205 Hi s Asn ser Ser Thr Glu Ser Phe lie Ala Asp Pro Ala lie Ser Leu 210 215 220 Ala Hi s Glu Leu lie His Ala Leu His Gly Leu Tyr Gly Ala Arg Gly 225 230 235 240 Val Thr Tyr Glu Glu Thr lie Glu val Lys Gin Ala Pro Leu Met lie 245 250 255 Ala Glu Lys Pro He Arg Leu Glu Glu Phe Leu Thr Phe Gly Gly Gin 260 265 270 Asp Leu Asn lie He Thr Ser Ala Met Lys Glu Lys He Tyr Asn Asn 275 280 285 Leu Leu Ala Asn Tyr Glu Lys lie Ala Thr Arg Leu Ser Glu val Asn 290 295 300 Ser Ala Pro pro Glu Tyr Asp He Asn Glu Tyr Lys Asp Tyr Phe Gin

    Page 117

    320 on

    Ο

    CN

    Ο <υ

    Ω kO

    P31OO3au-pct.txt

    315 vt m

    ο oo kO

    CN in o

    CN

    305 310

    Trp Lys Tyr Gly Leu 325 ASp Lys Asn Glu Asn Lys Phe 340 Asn Glu lie Tyr Ser Asp Leu 355 Ala Asn Lys Phe Lys 360 lie Lys 370 Tyr Glu Phe Leu Lys 375 Val Tyr 385 Thr val Ser Glu Gly 390 Phe Asn Arg Gly Gln Ser lie 405 Lys Leu Asn Asp Lys Gly Leu 420 Val Glu Lys He Asn Asn Asn 435 Asn Asn Asn Asn Asn 440 Thr ser 450 Lys Thr Lys Ser Leu 455 lie Met 465 Glu Val Ser cys Pro 470 A5p Gly Gly Ala Trp Gly cys 485 cys Pro Phe Hi s Ile Hi s Cys 500 Cys Pro Ala Gly Thr cys Glu 515 Ala Leu Ala Gly Gly 520 Gly Gly 530 Gly Gly ser Ala Leu 535 Val Ser 545 Glu Leu Phe Phe val 550 Ala Ser He Asn Thr Pro Lys 565 Glu lie Asp Tyr Glu Asn Asn Leu Asp Glu val

    Ala ASp 330 Gly ser Tyr Thr val 335 Asn Lys 345 Lys Leu Tyr Ser Phe 350 Thr Glu Val Lys Ala Arg Asn 365 Thr Tyr Phe Pro Asn Leu Leu 380 Asp Asp Asp Ile ile Gly Asn 395 Leu Ala Val Asn Asn 400 Pro Lys 410 Ile Ile Asp ser Ile 415 pro val 425 Lys Phe Ala val Glu 430 Asn Asn Leu Gly cys val Asp 445 Gly lie lie Glu Gly Arg Asp 460 val Lys cys Asp Tyr Thr cys 475 cys Arg Leu Gln ser 480 Thr Gln 490 Ala val Cys cys Glu 495 Asp Phe 505 Thr Cys Asp Thr Gln 510 Lys Gly Gly Gly Ser Gly Gly 525 Gly Gly Ser Leu Gln Cys lie 540 Glu val Asn Asn Glu Ser Ser 555 Tyr Asn Glu Asn Asp 560 Asp Thr 570 Thr Asn Leu Asn Asn 575 Asn ile Leu Asp Tyr Asn ser Gln Thr

    Page 118

    2015268754 16 Dec 2015 p31OO3au-pct.txt

    580 585 590

    He Pro Gln Ile Ser Asn Ile Glu Asn Leu Asn Thr Leu Val Gln Asp 595 600 605 Asn ser 610 Tyr Val Pro Glu Tyr 615 Asp ser Asn Gly Thr 620 Ser Glu lie Glu Glu Tyr Asp val val Asp Phe Asn val Phe Phe Tyr Leu His Ala Gln 625 630 635 640 Lys Val Pro Glu Gly Glu Thr Asn lie Ser Leu Thr Ser Ser lie Asp 645 650 655 Thr Ala Leu Leu Glu Glu Ser Lys Asp lie Phe Phe Ser Ser Glu Phe 660 665 670 lie Asp Thr Ile Asn Lys Pro val Asn Ala Ala Leu Phe lie Asp Trp 675 680 685 Ile Ser Lys val lie Arg Asp Phe Thr Thr Glu Ala Thr Gln Lys Ser 690 695 700 Thr Val Asp Lys lie Ala Asp lie Ser Leu Ile Val Pro Tyr Val Gly 705 710 715 720 Leu Ala Leu Asn lie lie lie Glu Ala Glu Lys Gly Asn Phe Glu Glu 725 730 735 Ala Phe Glu Leu Leu Gly Val Gly lie Leu Leu Glu Phe val Pro Glu 740 745 750 Leu Thr Ile Pro Val lie Leu val Phe Thr Ile Lys Ser Tyr ile Asp 755 760 765 ser Tyr Glu Asn Lys Asn Lys Ala Ile Lys Ala lie Asn Asn Ser Leu 770 775 780 lie Glu Arg Glu Ala Lys Trp Lys Glu lie Tyr Ser Trp lie val Ser 785 790 795 800 Asn Trp Leu Thr Arg ile Asn Thr Gln Phe Asn Lys Arg Lys Glu Gln 805 810 815 Met Tyr Gln Ala Leu Gln Asn Gln val Asp Ala lie Lys Thr Ala lie 820 825 830 Glu Tyr Lys Tyr Asn Asn Tyr Thr Ser Asp Glu Lys Asn Arg Leu Glu 835 840 845 ser Glu Tyr Asn lie Asn Asn Ile Glu Glu Glu Leu Asn Lys Lys val

    Page 119

    2015268754 16 Dec 2015

    P31OO3au-pct. txt 860

    850

    855

    ser Leu 865 Ala Met Lys Asn lie Glu 870 Arg Phe Met Thr Glu 875 Ser Ser lie 880 ser Tyr Leu Met Lys Leu lie Asn Glu Ala Lys Val Gly Lys Leu Lys 885 890 895 Lys Tyr Asp Asn His val Lys ser Asp Leu Leu Asn Tyr lie Leu Asp 900 905 910 Hi s Arg Ser lie Leu Gly Glu Gin Thr Asn Glu Leu ser Asp Leu Val 915 920 925 Thr Ser Thr Leu Asn Ser Ser lie Pro Phe Glu Leu ser ser Tyr Thr 930 935 940 Asn Asp Lys lie Leu lie lie Tyr Phe Asn Arg Leu Tyr Lys Thr 945 950 955

    <210> 40 <211> 939 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide

    <400> 40 Lys Asp phe Asn Tyr Ser Asp Pro Val Asn Asp Asn 15 Thr 1 Trp Pro Val 5 10 Asp He Leu Tyr Leu Arg lie pro Gin Asn Lys Leu He Thr Thr Pro 20 25 30 val Lys Ala Phe Met lie Thr Gin Asn He Trp val lie Pro Glu Arg 35 40 45 Phe ser Ser Asp Thr Asn Pro ser Leu Ser Lys Pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gin ser Tyr Tyr Asp Pro ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 Gin Lys Asp Thr Phe Leu Lys Gly He He Lys Leu Phe Lys Arg He 85 90 95 Asn Glu Arg Asp lie Gly Lys Lys Leu lie Asn Tyr Leu val val Gly 100 105 110 Ser pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125

    Page 120

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Thr Arg 130 His Thr Thr Asn Ile Ala 135 val Glu Lys Phe Glu Asn 140 Gly Ser Trp Lys val Thr Asn Ile Ile Thr pro Ser val Leu lie Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu Asp Tyr Thr Ala ser Leu Thr Leu Gin Gly Gin 165 170 175 Gin ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu Lys 180 185 190 val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp val Thr Ser Asn Gin 195 200 205 ser ser Ala val Leu Gly Lys Ser Ile Phe Cys Met Asp Pro val Ile 210 215 220 Ala Leu Met Hi s Glu Leu Thr Hi s ser Leu Hi s Gin Leu Tyr Gly lie 225 230 235 240 Asn Ile Pro Ser Asp Lys Arg lie Arg Pro Gin val Ser Glu Gly Phe 245 250 255 Phe Ser Gin Asp Gly Pro Asn Val Gin Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu Asp Val Glu ile Ile Pro Gin ile Glu Arg Ser Gin Leu 275 280 285 Arg Glu Lys Ala Leu Gly His Tyr Lys Asp He Ala Lys Arg Leu Asn 290 295 300 Asn lie Asn Lys Thr Ile pro Ser Ser Trp lie Ser Asn He Asp Lys 305 310 315 320 Tyr Lys Lys lie Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Gly Asn Phe val val Asn lie Asp Lys Phe Asn Ser Leu Tyr Ser Asp 340 345 350 Leu Thr Asn Val Met ser Glu Val Val Tyr Ser Ser Gin Tyr Asn val 355 360 365 Lys Asn Arg Thr His Tyr Phe Ser Arg Hi s Tyr Leu Pro val Phe Ala 370 375 380 Asn ile Leu Asp Asp Asn Ile Tyr Thr lie Arg ASP Gly Phe Asn Leu 385 390 395 400

    Page 121

    P31OO3au-pct.txt

    IT)

    Ο (Μ ο

    <υ

    Ω ^ιtn οο (Μ

    ΙΤ) ο

    (Μ

    Thr Asn Lys Gly Phe Asn Ile Glu Asn ser Gly Gin Asn Ile Glu Arg 405 410 415 Asn Pro Ala Leu Gin Lys Leu ser ser Glu Ser Val val Asp Leu Phe 420 425 430 Thr Lys val Cys Val Asp Gly Gly Gly Gly ser Gly Gly Gly Gly ser 435 440 445 Ala Asp Asp Asp Asp Lys Val val Ser Hi s Phe Asn Asp Cys Pro Asp 450 455 460 ser Hi s Thr Gin Phe Cys Phe Hi s Gly Thr Cys Arg phe Leu Val Gin 465 470 475 480 Glu Asp Lys pro Ala Cys val cys Hi s Ser Gly Tyr val Gly Ala Arg 485 490 495 cys Glu His Ala Asp Leu Leu Ala Leu Ala Gly Gly Gly Gly ser Gly 500 505 510 Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Ala Leu Gin cys lie 515 520 525 Lys Val Lys Asn Asn Arg Leu Pro Tyr Val Ala Asp Lys Asp Ser ile 530 535 540 Ser Gin Glu lie Phe Glu Asn Lys ile ile Thr Asp Glu Thr Asn val 545 550 555 560 Gin Asn Tyr Ser Asp Lys Phe Ser Leu Asp Glu Ser Ile Leu Asp Gly 565 570 575 Gin val Pro Ile Asn Pro Glu Ile val Asp Pro Leu Leu Pro Asn Val 580 585 590 Asn Met Glu Pro Leu Asn Leu Pro Gly Glu Glu lie val phe Tyr Asp 595 600 605 Asp ile Thr Lys Tyr Val Asp Tyr Leu Asn Ser Tyr Tyr Tyr Leu Glu 610 615 620 Ser Gin Lys Leu Ser Asn Asn val Glu Asn lie Thr Leu Thr. Thr ser 625 630 635 640 Val Glu Glu Ala Leu Gly Tyr ser Asn Lys lie Tyr Thr phe Leu Pro 645 650 655 Ser Leu Ala Glu Lys Val Asn Lys Gly Val Gin Ala Gly Leu Phe Leu

    660 665 670

    Page 122

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Asn Trp Ala 675 Asn Glu val Val Glu 680 Asp Phe Thr Thr Asn 685 ile Met Lys Lys Asp Thr Leu Asp Lys lie Ser Asp Val Ser Val ile lie Pro Tyr 690 695 700 lie Gly Pro Ala Leu Asn Ile Gly Asn Ser Ala Leu Arg Gly Asn Phe 705 710 715 720 Asn Gin Ala Phe Ala Thr Ala Gly Val Ala Phe Leu Leu Glu Gly Phe 725 730 735 Pro Glu Phe Thr Ile Pro Ala Leu Gly Val Phe Thr Phe Tyr Ser Ser 740 745 750 lie Gin Glu Arg Glu Lys Ile ile Lys Thr lie Glu Asn Cys Leu Glu 755 760 765 Gin Arg Val Lys Arg Trp Lys Asp ser Tyr Gin Trp Met Val Ser Asn 770 775 780 Trp Leu Ser Arg lie Thr Thr Gin Phe Asn Hi s Ile Asn Tyr Gin Met 785 790 795 800 Tyr Asp Ser Leu Ser Tyr Gin Ala Asp Ala lie Lys Ala Lys lie Asp 805 810 815

    Leu Glu Tyr Lys Lys Tyr Ser 820 Gly Ser 825 Asp Lys Glu Asn ile 830 Lys Ser Gin Val Glu Asn Leu Lys Asn Ser Leu Asp Val Lys Ile Ser Glu Ala 835 840 845 ( Met Asn Asn Ile Asn Lys Phe Ile Arg Glu Cys Ser Val Thr Tyr Leu 850 855 860 Phe Lys Asn Met Leu Pro Lys Val lie Asp Glu Leu Asn Lys Phe Asp 865 870 875 880 Leu Arg Thr Lys Thr Glu Leu Ile Asn Leu ile Asp Ser Hi s Asn lie 885 890 895 lie Leu val Gly Glu Val Asp Arg Leu Lys Ala Lys val Asn Glu ser 900 905 910 Phe Glu Asn Thr Met Pro Phe Asn Ile Phe Ser Tyr Thr Asn Asn Ser 915 920 925 Leu Leu Lys Asp lie lie Asn Glu Tyr Phe Asn 930 935

    Page 123

    P31OO3au-pct.txt

    754 16 Dec 2015 <210> 41 <211> 1002 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 41

    Thr Trp Pro Val Lys Asp Phe Asn Tyr Ser Asp Pro val Asn Asp Asn 15 10 15

    Asp lie Leu Tyr Leu Arg lie Pro Gln Asn Lys Leu lie Thr Thr Pro 20 25 30

    Val Lys Ala Phe Met lie Thr Gln Asn lie Trp Val Ile 45 Pro Glu Arg 35 40 Phe Ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro Thr 50 55 60 Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Gl u 65 70 75 80 Gln Lys Asp Thr Phe Leu Lys Gly Ile lie Lys Leu Phe Lys Arg He 85 90 95 Asn Glu Arg Asp lie Gly Lys Lys Leu ile Asn Tyr Leu val Val Gly 100 105 110 Ser Pro Phe Met Gly Asp Ser Ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg Hi s Thr Thr Asn lie Ala val Glu Lys Phe Glu Asn Gly Ser 130 135 140 Trp Lys Val Thr Asn Ile ile Thr Pro ser val Leu Ile Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu Asp Tyr Thr Ala ser Leu Thr Leu Gln Gly Gln 165 170 175 Gln ser Asn Pro Ser Phe Glu Gly Phe Gly Thr Leu Ser Ile Leu Lys 180 185 190 Val Al a Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr ser Asn Gln 195 200 205 ser ser Ala val Leu Gly Lys ser ile Phe cys Met Asp Pro Val lie 210 215 220

    Page 124

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    2015268754 16 Dec 2015

    Ala Leu Met His Glu Leu Thr His Ser Leu His Gl n Leu Tyr Gly lie 225 230 235 240 Asn lie Pro ’ Ser Asp Lys Arg lie Arg Pro Gin Val Ser Glu Gly Phe 245 250 255 Phe ser Gin Asp Gly Pro Asn Val Gin Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu Asp val Glu lie lie Pro Gin lie Gl u Arg Ser Gin Leu 275 280 285 Arg Glu Lys Ala Leu Gly His Tyr Lys Asp lie Ala Lys Arg Leu Asn 290 295 300 Asn lie Asn Lys Thr lie Pro Ser Ser Trp lie Ser Asn lie Asp Lys 305 310 315 320 Tyr Lys Lys lie Phe Ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Gly Asn Phe Val Val Asn lie Asp Lys Phe Asn ser Leu Tyr Ser Asp 340 345 350 Leu Thr Asn val Met Ser Glu val Val Tyr Ser ser Gin Tyr Asn Val 355 360 365 Lys Asn Arg Thr Hi s Tyr Phe Ser Arg His Tyr Leu Pro val phe Ala 370 375 380 Asn lie Leu Asp Asp Asn lie Tyr Thr lie Arg Asp Gly Phe Asn Leu 385 390 395 400 Thr Asn Lys Gly Phe Asn lie Glu Asn Ser Gly Gin Asn lie Glu Arg 405 410 415 Asn Pro Ala Leu Gin Lys Leu Ser Ser Glu Ser val val Asp Leu Phe 420 425 430 Thr Lys val Cys Val Asp Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 435 440 445 Ala Asp Asp Asp Asp Lys Ala Leu Asp Thr Asn Tyr Cys Phe ser Ser 450 455 460 Thr Glu Lys Asn Cys Cys val Arg Gin Leu Tyr lie Asp Phe Arg Lys 465 470 475 480 Asp Leu Gly Trp Lys Trp lie Hi s Glu Pro Lys Gly Tyr His Ala Asn 485 490 495

    Page 125

    P31OO3au-pct.txt

    Ο (Μ Phe cys Leu Gly Pro Cys Pro Tyr Ile Trp ser Leu Asp Thr Gin Tyr 500 505 510 Ο <D Ser Lys Val Leu Ala Leu Tyr Asn Gin His Asn Pro Gly Ala Ser Ala Ω 515 520 525 Ala Pro cys cys val Pro Gin Ala Leu Glu Pro Leu Pro lie val Tyr 530 535 540 Tl- V Tyr val Gly Arg Lys Pro Lys val Glu Gin Leu ser Asn Met He val OO <N 545 550 555 560 Arg Ser Cys Lys Cys Ser Ala Leu Ala Gly Gly Gly Gly Ser Gly Gly V 565 570 575 o Ala Gin lie <N Gly Gly Ser Gly Gly 580 Gly Gly Ser Ala 585 Leu Leu cys 590 Lys Val Lys Asn Asn Arg Leu Pro Tyr Val Ala Asp Lys Asp Ser lie Ser 595 600 605 Gin Glu lie Phe Glu Asn Lys lie lie Thr Asp Glu Thr Asn Val Gin 610 615 620 Asn Tyr Ser Asp Lys Phe Ser Leu Asp Glu Ser lie Leu Asp Gly Gin 625 630 635 640 Val Pro ile Asn Pro Glu ile val Asp Pro Leu Leu Pro Asn val Asn 645 650 655 Met Glu Pro Leu Asn Leu Pro Gly Glu Glu ile Val Phe Tyr Asp Asp 660 665 670 Ile Thr Lys Tyr Val Asp Tyr Leu Asn Ser Tyr Tyr Tyr Leu Glu Ser 675 680 685 Gin Lys Leu Ser Asn Asn Val Glu Asn ile Thr Leu Thr Thr ser Val 690 695 700 Glu Glu Ala Leu Gly Tyr Ser Asn Lys lie Tyr Thr Phe Leu Pro Ser 705 710 715 720 Leu Ala Glu Lys val Asn Lys Gly val Gin Ala Gly Leu Phe Leu Asn 725 730 735 Trp Ala Asn Glu val val Glu Asp Phe Thr Thr Asn Ile Met Lys Lys 740 745 750 Asp Thr Leu Asp Lys lie Ser Asp val ser Val lie He Pro Tyr Ile 755 760 765

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    P31OO3au-pct. txt

    2015268754 16 Dec 2015

    Gly Pro 770 Ala Leu Asn lie Gly Asn 775 Ser Ala Leu Arg 780 Gly Asn Phe Asn Gin Ala Phe Ala Thr Ala Gly val Ala Phe Leu Leu Glu Gly Phe Pro 785 790 795 800 Glu Phe Thr lie Pro Ala Leu Gly val Phe Thr Phe Tyr Ser ser lie 805 810 815 Gin Glu Arg Glu Lys lie lie Lys Thr lie Glu Asn cys Leu Glu Gin 820 825 830 Arg Val Lys Arg Trp Lys Asp Ser Tyr Gin Trp Met Val Ser Asn Trp 835 840 845 Leu Ser Arg lie Thr Thr Gin Phe Asn Hi s lie Asn Tyr Gin Met Tyr 850 855 860 Asp Ser Leu Ser Tyr Gin Ala Asp Ala lie Lys Ala Lys He Asp Leu 865 870 875 880 Glu Tyr Lys Lys Tyr Ser Gly Ser Asp Lys Glu Asn lie Lys Ser Gin 885 890 895 Val Glu Asn Leu Lys Asn Ser Leu Asp val Lys lie Ser Glu Ala Met 900 905 910 Asn Asn lie Asn Lys Phe lie Arg Glu Cys Ser val Thr Tyr Leu Phe 915 920 925 Lys Asn Met Leu Pro Lys Val lie Asp Glu Leu Asn Lys Phe Asp Leu 930 935 940 Arg Thr Lys Thr Glu Leu lie Asn Leu lie Asp Ser His Asn lie lie 945 950 955 960 Leu val Gly Glu Val Asp Arg Leu Lys Al a Lys Val Asn Glu Ser Phe 965 970 975 Glu Asn Thr Met Pro Phe Asn lie Phe Ser Tyr Thr Asn Asn Ser Leu

    980 985 990

    Leu Lys Asp lie lie Asn Glu Tyr Phe Asn 995 1000 <220>

    <221> source <210> 42 <211> 1045 <212> PRT <213> Artificial sequence

    Page 127

    P31OO3au-pct.txt <223> /note=Description of Artificial sequence: synthetic θ polypeptide ^0x1 <400> 42

    O Pro val Thr lie Asn Asn Phe Asn Tyr Asn Asp Pro Ile 10 Asp Asn Asn 15 i Q Met 20 5 Met Glu Pro Pro Phe 25 Ala Arg Gly Thr Asn ile lie Gly 30 Arg Tyr T-t- Tyr Lys Ala Phe Lys Ile Thr Asp Arg lie Trp Ile lie Pro Glu Arg •ΖΊ 35 40 45 l> oo Tyr Thr Phe Gly Tyr Lys Pro Glu Asp phe Asn Lys ser Ser Gly Ile 50 55 60 vn r-H Phe Asn Arg Asp val Cys Glu Tyr Tyr Asp Pro Asp Tyr Leu Asn Thr <N 65 70 75 80 Asn Asp Lys Lys Asn Ile Phe Leu Gin Thr Met lie Lys Leu phe Asn 85 90 95 Arg ile Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met ile lie 100 105 110 Asn Gly ile Pro Tyr Leu Gly Asp Arg Arg Val Pro Leu Glu Glu Phe 115 120 125 Asn Thr Asn Ile Ala Ser val Thr Val Asn Lys Leu ile Ser Asn Pro 130 135 140 Gly Glu val Glu Arg Lys Lys Gly Ile Phe Ala Asn Leu ile lie Phe 145 150 155 160 Gly Pro Gly Pro Val Leu Asn Glu Asn Glu Thr ile Asp ile Gly lie 165 170 175 Gin Asn Hi s Phe Ala Ser Arg Glu Gly Phe Gly Gly Ile Met Gin Met 180 185 190 Lys Phe Cys Pro Glu Tyr val Ser Val Phe Asn Asn val Gin Glu Asn 195 200 205 Lys Gly Ala Ser lie Phe Asn Arg Arg Gly Tyr Phe Ser Asp Pro Ala 210 215 220 Leu ile Leu Met Hi s Glu Leu Ile Hi s val Leu Hi s Gly Leu Tyr Gly 225 230 235 240 Ile Lys val Asp Asp Leu Pro lie val Pro Asn Glu Lys Lys Phe Phe 245 250 255

    Page 128

    2015268754 16 Dec 2015

    P31 003a u-pc t.tx t Met Gin Ser Thr Asp Ala Ile Gin Ala Glu Glu Leu Tyr Thr Phe Gly 260 265 270 Gly Gin Asp Pro Ser lie lie Thr Pro Ser Thr Asp Lys Ser lie Tyr 275 280 285 Asp Lys val Leu Gin Asn Phe Arg Gly lie val Asp Arg Leu Asn Lys 290 295 300 val Leu Val Cys lie Ser Asp Pro Asn lie Asn lie Asn Ile Tyr Lys 305 310 315 320 Asn Lys Phe Lys Asp Lys Tyr Lys Phe Val Glu Asp Ser Glu Gly Lys 325 330 335 Tyr Ser Ile Asp val Glu Ser Phe Asp Lys Leu Tyr Lys Ser Leu Met 340 345 350 Phe Gly Phe Thr Glu Thr Asn lie Ala Glu Asn Tyr Lys Ile Lys Thr 355 360 365 Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro val Lys Ile Lys Asn 370 375 380 Leu Leu Asp Asn Glu lie Tyr Thr Ile Glu Glu Gly Phe Asn Ile Ser 385 390 395 400 Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gin Asn Lys Ala lie Asn 405 410 415 Lys Gin Ala Tyr Glu Glu lie Ser Lys Glu His Leu Ala Val Tyr Lys 420 425 430 lie Gin Met Cys Val Asp Glu Glu Lys Leu Tyr Asp Asp Asp Asp Lys 435 440 445 Asp Arg Trp Gly Ser Ser Leu Gin Cys lie Asp val Asp Asn Glu Asp 450 455 460 Leu Phe Phe lie Ala Asp Lys Asn Ser Phe ser Asp Asp Leu Ser Lys 465 470 475 480 Asn Glu Arg lie Glu Tyr Asn Thr Gin ser Asn Tyr lie Glu Asn Asp 485 490 495 Phe Pro Ile Asn Glu Leu Ile Leu Asp Thr Asp Leu lie Ser Lys lie 500 505 510 Glu Leu Pro Ser Glu Asn Thr Glu Ser Leu Thr Asp Phe Asn val Asp 515 520 525

    Page 129

    P31OO3au-pct. txt

    ^Val O CH Pro 530 Asn Arg val Tyr Glu Lys Gin Pro Ala lie Lys Lys lie Phe Thr Asp 540 Thr lie 535 Phe Gin Tyr 550 Leu Tyr ser Gin 555 Thr Phe Pro Leu Asp 560 o <D Ω 80 Glu 545 lie Asp lie Ser 565 Leu Thr Ser Ser Phe 570 Asp Asp Ala Leu Leu 575 Phe Ser Asn Lys val Tyr ser Phe Phe ser Met Asp Tyr lie Lys Thr Ala XT ΜΊ 580 585 590 OO Asn Lys val Val Glu Ala Gly Leu Phe Ala Gly Trp Val Lys Gin lie ΓΗ 595 600 605 IT) val Asn ASP Phe val lie Glu Ala Asn Lys Ser Asn Thr Met Asp Al a ΓΗ 610 615 620 He Ala Asp lie Ser Leu lie Val Pro Tyr lie Gly Leu Ala Leu Asn 625 630 635 640 val Gly Asn Glu Thr Ala Lys Gly Asn Phe Glu Asn Ala Phe Glu lie 645 650 655 Ala Gly Ala ser lie Leu Leu Glu Phe lie Pro Glu Leu Leu lie Pro 660 665 670 val val Gly Ala Phe Leu Leu Glu Ser Tyr lie Asp Asn Lys Asn Lys 675 680 685 lie lie Lys Thr He Asp Asn Ala Leu Thr Lys Arg Asn Glu Lys Trp 690 695 700 Ser Asp Met Tyr Gly Leu lie Val Ala Gin Trp Leu ser Thr Val Asn 705 710 715 720 Thr Gin Phe Tyr Thr lie Lys Glu Gly Met Tyr Lys Ala Leu Asn Tyr 725 730 735 Gin Ala Gin Ala Leu Glu Glu lie lie Lys Tyr Arg Tyr Asn lie Tyr 740 745 750 Ser Glu Lys Glu Lys Ser Asn He Asn lie Asp Phe Asn Asp lie Asn 755 760 765 Ser Lys Leu Asn Glu Gly lie Asn Gin Ala lie Asp Asn lie Asn Asn 770 775 780 Phe lie Asn Gly cys ser Val Ser Tyr Leu Met Lys Lys Met lie Pro 785 790 795 800

    Page 130

    P31003au-pct.txt

    2015268754 16 Dec 2015

    Leu Ala val Glu Lys 805 Leu Leu Asp Phe Asp 810 Asn Thr Leu Lys Lys 815 Asn Leu Leu Asn Tyr Ile Asp Glu Asn Lys Leu Tyr Leu Ile Gly ser Ala 820 825 830 Glu Tyr Glu Lys Ser Lys val Asn Lys Tyr Leu Lys Thr Ile Met Pro 835 840 845 Phe Asp Leu Ser lie Tyr Thr Asn Asp Thr lie Leu Ile Glu Met Phe 850 855 860 Asn Lys Tyr Asn Ser Leu Glu Gly Gly Gly Gly Ser Gly Gly Gly Gly 865 870 875 880 ser Gly Gly Gly Gly ser Ala Leu Val Arg Ser Ser Ser Arg Thr pro 885 890 895 Ser Asp Lys Pro Val Ala His Val val Ala Asn Pro Gln Ala Glu Gly 900 905 910 Gln Leu Gln Trp Leu Asn Arg Arg Ala Asn Ala Leu Leu Ala Asn Gly 915 920 925 Val Glu Leu Arg Asp Asn Gln Leu val Val Pro ser Glu Gly Leu Tyr 930 935 940 Leu lie Tyr ser Gln val Leu Phe Lys Gly Gln Gly cys Pro Ser Thr 945 950 955 960 Hi s val Leu Leu Thr His Thr Ile Ser Arg Ile Ala Val Ser Tyr Gln 965 970 975 Thr Lys val Asn Leu Leu Ser Ala lie Lys Ser Pro Cys Gln Arg Glu 980 985 990 Thr Pro Glu Gly Ala Glu Ala Lys Pro * Trp Tyr Glu Pro i ile Tyr Leu 995 1000 1005 Gly Gly val Phe Gln Leu Glu Lys Gly Asp Arg Leu Ser Ala Glu 1010 1015 1020 lie Asn Arg Pro Asp Tyr Leu Asp Phe Ala Glu ser Gly Gln val 1025 1030 1035 Tyr Phe Gly ile lie Al a Leu 1040 1045

    <210> 43 <211> 962 <212> PRT <213> Artificial Sequence

    Page 131

    P31OO3au-pct. txt

    IT)

    T? <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic O polypeptide <D

    Ω <400> 43 Thr Trp Pro 1 val Lys 5 Asp phe Asn Tyr Ser 10 Asp Pro val Asn Asp 15 Asn Asp Ile Leu Tyr Leu Arg lie Pro Gln Asn Lys Leu Ile Thr Thr Pro ^1^- 20 25 30 IT) O val Lys Ala Phe Met lie Thr Gln Asn lie Trp Val lie Pro Glu Arg 35 40 45 <N VD Phe ser Ser Asp Thr Asn Pro Ser Leu Ser Lys Pro Pro Arg Pro Thr O 50 55 60 (N Ser Lys Tyr Gln Ser Tyr Tyr Asp Pro Ser Tyr Leu Ser Thr Asp Glu 65 70 75 80 Gln Lys Asp Thr phe Leu Lys Gly lie lie Lys Leu Phe Lys Arg lie 85 90 95 Asn Glu Arg Asp Ile Gly Lys Lys Leu lie Asn Tyr Leu val Val Gly 100 105 110 Ser Pro Phe Met Gly Asp ser ser Thr Pro Glu Asp Thr Phe Asp Phe 115 120 125 Thr Arg His Thr Thr Asn lie Ala Val Glu Lys Phe Glu Asn Gly Ser 130 135 140 Trp Lys val Thr Asn Ile Ile Thr Pro Ser val Leu ile Phe Gly Pro 145 150 155 160 Leu Pro Asn lie Leu Asp Tyr Thr Ala Ser Leu Thr Leu Gln Gly Gln 165 170 175 Gln ser Asn Pro ser Phe Glu Gly Phe Gly Thr Leu Ser lie Leu Lys 180 185 190 val Ala Pro Glu Phe Leu Leu Thr Phe Ser Asp Val Thr Ser Asn Gln 195 200 205 Ser Ser Ala val Leu Gly Lys Ser lie Phe cys Met ASp Pro val lie 210 215 220 Ala Leu Met Hi s Glu Leu Thr Hi s ser Leu His Gln Leu Tyr Gly Ile 225 230 235 240 Asn Ile Pro Ser Asp Lys Arg lie Arg Pro Gln val Ser Glu Gly Phe

    Page 132

    2015268754 16 Dec 2015

    245 P31OO3au-pct.txt 250 255 Phe Ser Gin Asp Gly pro Asn Val Gin Phe Glu Glu Leu Tyr Thr Phe 260 265 270 Gly Gly Leu Asp val Glu lie lie Pro Gin lie Glu Arg Ser Gin Leu 275 280 285 Arg Glu 290 Lys Ala Leu Gly His 295 Tyr Lys Asp lie Ala 300 Lys Arg Leu Asn Asn ile Asn Lys Thr Ile Pro Ser Ser Trp lie Ser Asn Ile Asp Lys 305 310 315 320 Tyr Lys Lys He Phe ser Glu Lys Tyr Asn Phe Asp Lys Asp Asn Thr 325 330 335 Gly Asn Phe val val Asn lie Asp Lys Phe Asn Ser Leu Tyr Ser Asp 340 345 350 Leu Thr Asn Val Met Ser Glu val Val Tyr ser ser Gin Tyr Asn val 355 360 365 Lys Asn Arg Thr Hi s Tyr Phe Ser Arg His Tyr Leu Pro val Phe Ala 370 375 380 Asn lie Leu Asp ASp Asn lie Tyr Thr lie Arg Asp Gly Phe Asn Leu 385 390 395 400 Thr Asn Lys Gly Phe Asn Ile Glu Asn Ser Gly Gin Asn lie Glu Arg 405 410 415 Asn Pro Ala Leu Gin Lys Leu Ser Ser Glu Ser Val val Asp Leu Phe 420 425 430 Thr Lys Val Cys val Asp Lys Ser Glu Glu Lys Leu Tyr Asp Asp Asp 435 440 445 Asp Lys Asp Arg Trp Gly Ser Ser Leu Gin cys lie Lys Val Lys Asn 450 455 460 Asn Arg Leu Pro Tyr Val Ala Asp Lys Asp ser Ile ser Gin Glu Ile 465 470 475 480 Phe Glu Asn Lys lie Ile Thr Asp Glu Thr Asn val Gin Asn Tyr Ser 485 490 495 Asp Lys Phe Ser Leu Asp Glu Ser Ile Leu Asp Gly Gin Val Pro lie 500 505 510 Asn Pro Glu lie val Asp Pro Leu Leu Pro Asn val Asn Met Glu Pro Page i 133

    P31OO3au-pct.txt

    515 520 525

    Ο

    CM Leu Asn Leu Pro Gly Glu Glu lie Val Phe Tyr Asp Asp lie Thr Lys Ο 530 535 540 <υ ο Ω Tyr Val Asp Tyr Leu Asn Ser Tyr Tyr Tyr Leu Glu Ser Gin Lys Leu 545 550 555 560 Ser Asn Asn val Glu Asn lie Thr Leu Thr Thr Ser Val Glu Glu Ala τψ 565 570 575 ΙΤ) Ο 00 Leu Gly Tyr Ser Asn Lys lie Tyr Thr Phe Leu Pro Ser Leu Ala Glu 580 585 590 CM IT) Lys Val Asn Lys Gly val Gin Ala Gly Leu Phe Leu Asn Trp Ala Asn ο 595 600 605 CM Glu val val Glu Asp Phe Thr Thr Asn lie Met Lys Lys Asp Thr Leu 610 615 620 Asp Lys lie Ser Asp val Ser Val lie lie Pro Tyr lie Gly Pro Ala 625 630 635 640 Leu Asn lie Gly Asn ser Ala Leu Arg Gly Asn phe Asn Gin Ala Phe 645 650 655 Al a Thr Ala Gly Val Ala Phe Leu Leu Glu Gly Phe Pro Glu Phe Thr 660 665 670 lie Pro Ala Leu Gly val Phe Thr Phe Tyr Ser Ser lie Gin Glu Arg 675 680 685 Glu Lys lie lie Lys Thr lie Glu Asn Cys Leu Gl u Gin Arg Val Lys 690 695 700 Arg Trp Lys Asp ser Tyr Gin Trp Met Val Ser Asn Trp Leu ser Arg 705 710 715 720 lie Thr Thr Gin Phe Asn Hi s lie Asn Tyr Gin Met Tyr Asp Ser Leu 725 730 735 Ser Tyr Gin Ala Asp Ala lie Lys Ala Lys lie ASp Leu Glu Tyr Lys 740 745 750 Lys Tyr Ser Gly Ser Asp Lys Glu Asn lie Lys ser Gin Val Glu Asn 755 760 765 Leu Lys Asn Ser Leu Asp val Lys lie Ser Glu Ala Met Asn Asn lie 770 775 780 Asn Lys Phe lie Arg Glu Cys Ser val Thr Tyr Leu phe Lys Asn Met

    Page 134

    2015268754 16 Dec 2015

    P31003au-pct.txt

    785 790 795 800

    Leu Pro Lys Val lie Asp 805 Glu Leu Asn Lys 810 Phe Asp Leu Arg Thr 815 Lys Thr Glu Leu lie Asn Leu lie Asp Ser Hi s Asn He He Leu val Gly 820 825 830 Glu val Asp Arg Leu Lys Ala Lys Val Asn Glu Ser Phe Glu Asn Thr 835 840 845 Met Pro Phe Asn lie Phe Ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp 850 855 860 lie Ile Asn Glu Tyr Phe Asn Leu Glu Gly Gly Gly Gly ser Gly Gly 865 870 875 880 Gly Gly ser Gly Gly Gly Gly Ser Ala Leu Lys Pro val ser Leu ser 885 890 895 Tyr Arg Cys Pro Cys Arg Phe Phe Glu Ser Hi s Val Ala Arg Ala Asn 900 905 910 val Lys His Leu Lys lie Leu Asn Thr Pro Asn Cys Ala Leu Gln ile 915 920 925 val Ala Arg Leu Lys Asn Asn Asn Arg Gln val cys lie Asp Pro Lys 930 935 940 Leu Lys Trp Ile Gln Glu Tyr Leu Glu Lys Ala Leu Asn Lys Arg Phe 945 950 955 960

    Lys Met (

    <210> 44 <211> 1090 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic polypeptide <400> 44

    Pro Ile Thr Ile Asn Asn Phe Asn Tyr Ser Asp pro val Asp Asn Lys 1 5 10 15 Asn lie Leu Tyr Leu Asp Thr Hi s Leu Asn Thr Leu Ala Asn Glu Pro 20 25 30 Glu Lys Ala Phe Arg ile Thr Gly Α5Π Ile Trp Val lie Pro Asp Arg

    35 40 45

    Page 135

    P31OO3au-pct.txt in r—Η

    Ο

    ΓΊ

    Ο ο

    Ω

    NO τΤ ιη ο

    οο

    NO

    ΓΊ ιη

    Ο

    ΓΊ

    Phe ser Arg Asn ser Asn 50 Pro Asn Leu Asn Lys Pro Pro Arg Val Thr 55 60 Ser Pro Lys ser Gly Tyr Tyr Asp Pro Asn Tyr Leu ser Thr ASp ser 65 70 75 80 Asp Lys Asp Thr Phe Leu Lys Glu lie lie Lys Leu Phe Lys Arg lie 85 90 95 Asn Ser Arg Glu Ile Gly Glu Glu Leu lie Tyr Arg Leu ser Thr Asp 100 105 110 He Pro Phe Pro Gly Asn Asn Asn Thr Pro lie Asn Thr Phe Asp Phe 115 120 125 Asp Val ASP Phe Asn ser Val Asp Val Lys Thr Arg Gln Gly Asn Asn 130 135 140 Trp val Lys Thr Gly ser ile Asn Pro Ser val Ile Ile Thr Gly Pro 145 150 155 160 Arg Glu Asn Ile lie Asp Pro Glu Thr Ser Thr Phe Lys Leu Thr Asn 165 170 175 Asn Thr Phe Ala Ala Gln Glu Gly Phe Gly Al a Leu Ser lie Ile Ser 180 185 190 He Ser Pro Arg phe Met Leu Thr Tyr Ser Asn Ala Thr Asn Asp val 195 200 205 Gly Glu Gly Arg Phe Ser Lys Ser Glu Phe Cys Met Asp Pro Ile Leu 210 215 220 lie Leu Met HIS Glu Leu Asn His Ala Met His Asn Leu Tyr Gly lie 225 230 235 240 Ala lie pro Asn Asp Gln Thr lie Ser Ser val Thr ser Asn Ile Phe 245 250 255 Tyr ser Gln Tyr Asn Val Lys Leu Glu Tyr Ala Glu Ile Tyr Ala Phe 260 265 270 Gly Gly pro Thr lie Asp Leu lie Pro Lys Ser Ala Arg Lys Tyr Phe 275 280 285 Glu Glu Lys Ala Leu Asp Tyr Tyr Arg Ser lie Ala Lys Arg Leu Asn 290 295 300 ser ile Thr Thr Ala Asn Pro Ser Ser Phe Asn Lys Tyr Ile Gly Gl u

    305 310 315 320

    Page 136

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Tyr Lys Gln Lys Leu ile Arg Lys Tyr Arg Phe val Val Glu Ser 335 Ser 325 330 Gly Glu val Thr val Asn Arg Asn Lys Phe Val Glu Leu Tyr Asn Glu 340 345 350 Leu Thr Gln Ile Phe Thr Glu Phe Asn Tyr Ala Lys lie Tyr Asn Val 355 360 365 Gln Asn Arg Lys Ile Tyr Leu ser Asn Val Tyr Thr Pro val Thr Ala 370 375 380 Asn Ile Leu Asp Asp Asn Val Tyr Asp lie Gln Asn Gly Phe Asn Ile 385 390 395 400 Pro Lys Ser Asn Leu Asn Val Leu Phe Met Gly Gln Asn Leu Ser Arg 405 410 415 Asn Pro Ala Leu Arg Lys val Asn Pro Glu Asn Met Leu Tyr Leu Phe 420 425 430 Thr Lys Phe Cys Val Asp Ala ile Asp Gly Arg Ser Leu Tyr Asn Lys 435 440 445 Thr Leu Gln Cys Arg Glu Leu Leu val Lys Asn Thr Asp Leu Pro Phe 450 455 460 Ile Gly Asp ile Ser Asp val Lys Thr Asp lie Phe Leu Arg Lys Asp 465 470 475 480 Ile Asn Glu Glu Thr Glu Val Ile Tyr Tyr pro Asp Asn Val Ser val 485 490 495 Asp Gln val lie Leu Ser Lys Asn Thr Ser Glu His Gly Gln Leu Asp 500 505 510 Leu Leu Tyr Pro Ser Ile Asp Ser Glu Ser Glu lie Leu pro Gly Glu 515 520 525 Asn Gln val Phe Tyr Asp Asn Arg Thr Gln Asn val Asp Tyr Leu Asn 530 535 540 ser Tyr Tyr Tyr Leu Glu ser Gln Lys Leu Ser Asp Asn val Glu Asp 545 550 555 560 Phe Thr Phe Thr Arg Ser lie Glu Glu Ala Leu ASp Asn Ser Ala Lys 565 570 575 Val Tyr Thr Tyr Phe Pro Thr Leu Ala Asn Lys val Asn Ala Gly val

    580 585 590

    Page 137

    P31OO3au-pct. txt in r—M o

    CH o

    CD

    Ω in oo kO <N in π-Η ©

    CM

    Gin Gly Gly Leu Phe Leu Met Trp Ala Asn Asp Val Val 605 Glu Asp Phe 595 600 Thr Thr Asn lie Leu Arg Lys Asp Thr Leu Asp Lys lie ser Asp val 610 615 620 Ser Ala lie lie Pro Tyr lie Gly Pro Ala Leu Asn lie ser Asn Ser 625 630 635 640 Val Arg Arg Gly Asn Phe Thr Glu Ala Phe Ala val Thr Gly val Thr 645 650 655 lie Leu Leu Glu Ala Phe Pro Glu Phe Thr lie Pro Ala Leu Gly Ala 660 665 670 Phe val lie Tyr Ser Lys val Gin Glu Arg Asn Glu lie Ile Lys Thr 675 680 685 lie Asp Asn Cys Leu Glu Gin Arg lie Lys Arg Trp Lys Asp Ser Tyr 690 695 700 Glu Trp Met Met Gly Thr Trp Leu ser Arg lie ile Thr Gin Phe Asn 705 710 715 720 Asn lie Ser Tyr Gin Met Tyr Asp ser Leu Asn Tyr Gin Ala Gly Ala 725 730 735 lie Lys Ala Lys lie Asp Leu Glu Tyr Lys Lys Tyr Ser Gly ser Asp 740 745 750 Lys Gl u Asn lie Lys Ser Gin Val Glu Asn Leu Lys Asn Ser Leu Asp 755 760 765 val Lys Ile Ser Glu Ala Met Asn Asn lie Asn Lys Phe ile Arg Glu 770 775 780 cys Ser val Thr Tyr Leu Phe Lys Asn Met Leu Pro Lys val lie Asp 785 790 795 800 Glu Leu Asn Glu Phe Asp Arg Asn Thr Lys Ala Lys Leu Ile Asn Leu 805 810 815 lie Asp Ser Hi s Asn lie lie Leu val Gly Glu val Asp Lys Leu Lys 820 825 830 Ala Lys val Asn Asn Ser Phe Gin Asn Thr ile Pro Phe Asn Ile Phe 835 840 845 ser Tyr Thr Asn Asn Ser Leu Leu Lys Asp lie lie Asn Glu Tyr Phe 850 855 860

    Page 138

    P31003au-pct.txt

    2015268754 16 Dec 2015

    Asn 865 Leu Glu Gly Gly Gly 870 Gly Ser Gly Gly Gly Gly Ser Gly Gly 875 Gly 880 Gly Ser Ala Leu Ala Pro Met Ala Glu Gly Gly Gly Gin Asn His His 885 890 895 Glu Val Val Lys Phe Met Asp Val Tyr Gin Arg Ser Tyr Cys His Pro 900 905 910 lie Glu Thr Leu Val Asp lie Phe Gin Glu Tyr Pro Asp Glu lie Glu 915 920 925 Tyr lie Phe Lys Pro Ser cys Val Pro Leu Met Arg Cys Gly Gly cys 930 935 940 Cys Asn Asp Glu Gly Leu Glu cys val Pro Thr Glu Glu Ser Asn lie 945 950 955 960 Thr Met Gl n lie Met Arg lie Lys Pro Hi s Gin Gly Gin Hi s lie Gly 965 970 975 Glu Met Ser Phe Leu Gin Hi s Asn Lys Cys Glu cys Arg Pro Lys Lys 980 985 990

    Asp Arg Ala Arg Gin Glu Lys Lys Ser val Arg Gly Lys Gly Lys Gly

    995 1000 1005 Gin Lys Arg Lys Arg Lys Lys Ser Arg Tyr Lys Ser Trp Ser Val 1010 1015 1020 Tyr Val Gly Ala Arg cys cys Leu Met Pro Trp Ser Leu Pro Gly 1025 1030 1035 Pro His Pro Cys Gly Pro cys ser Glu Arg Arg Lys His Leu Phe 1040 1045 1050 Val Gin Asp Pro Gin Thr Cys Lys Cys ser Cys Lys Asn Thr Asp 1055 1060 1065 Ser Arg cys Lys Ala Arg Gin Leu Glu Leu Asn Glu Arg Thr Cys 1070 1075 1080 Arg Cys Asp Lys Pro Arg Arg 1085 1090

    <210> 45 <211> 10 <212> PRT <213> Artificial sequence <220>

    Page 139

    P31OO3au~pct. txt

    2015268754 16 Dec 2015 <221> source <223> /note=Description of Artificial Sequence: synthetic peptide <220>

    <221> MOD_RES <222> (1)..(1) <223> pyro-Glutamate <400> 45

    Glu His Trp ser Tyr Gly Leu Arg Pro Gly 15 10 <210> 46 <211> 6 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Transforming growth factor alpha peptide <400> 46

    Trp Ser His Phe Asn Asp

    1 5 <210> 47 <211> 5 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Neuregulin peptide <400> 47

    Ser His Leu Val Lys

    1 5 <210> 48 <211> 5 <212> PRT <213> Unknown <22O>

    <221> source <223> /note=Description of unknown: Epidermal growth factor peptide <400> 48

    Asn Ser Asp ser Glu

    1 5 <210> 49 <211> 44 <212> PRT <213> Artificial sequence <22O>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide

    Page 140

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    <400> 49 Tyr Ala Asp Ala lie Phe Thr Asn Ser Tyr Arg Lys val Leu Gly Gin 1 5 10 15 Leu Ser Ala Arg Lys Leu Leu Gin Asp He Met Ser Arg Gin Gin Gly 20 25 30 Glu Ser Asn Gin Glu Arg Gly Ala Arg Ala Arg Leu 35 40

    <210> 50 <211> 29 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic peptide <400> 50

    Tyr Ala Asp Ala lie Phe Thr Asn Ser Tyr Arg Lys Val Leu Gly Gin 15 10 15

    Leu Ser Ala Arg Lys Leu Leu Gin Asp lie Met Ser Arg 20 25 <210> 51 <211> 44 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 51

    Tyr Ala Asp Ala lie Phe Thr Asn Ser Tyr Arg Lys lie Leu Gly Gin ( ¹ 5 10 15 Leu Ser Ala Arg Lys Leu Leu Gin Asp lie Met Asn Arg Gin Gin Gly 20 25 30 Glu Arg Asn Gin Glu Gin Gly Ala Lys val Arg Leu

    35 40 <210> 52 <211> 44 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 52

    Tyr Ala Asp Ala lie Phe Thr Asn Ser Tyr Arg Lys Val Leu Gly Gin Page 141

    2015268754 16 Dec 2015

    P31OO3au-pct.txt

    15 10 15

    Leu Ser Ala Arg Lys Leu Leu Gin Asp lie Met Asn Arg Gin Gin Gly 20 25 30

    Glu Arg Asn Gin Glu Gin Gly Ala Lys val Arg Leu 35 40 <210> 53 <211> 29 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic peptide <400> 53

    His val Asp Ala lie Phe Thr Gin ser Tyr Arg Lys Val Leu Ala Gin 15 10 15

    Leu ser Ala Arg Lys Leu Leu Gin Asp lie Leu Asn Arg 20 25 <210> 54 <211> 40 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic polypeptide <400> 54

    His 1 val Asp Ala lie Phe Thr Gin ser Tyr Arg Lys val Leu Ala 15 Gin 5 10 Leu ser Ala Arg Lys Leu Leu Gin Asp lie Leu Asn Arg Gin Gin Gly 20 25 30 Glu Arg Asn Gin Glu Gin Gly Al a

    35 40 <210> 55 <211> 14 <212> PRT <213> Unknown <22O>

    <221> source <223> /note=Description of unknown: Frog bombesin peptide <220>

    <221> MOD_RES <222> (1)..(1) <223> pyro-Glutamate

    Page 142

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <400> 55

    Glu Gin Arg Leu Gly Asn Gin Trp Ala Val Gly His Leu Met 15 10 <210> 56 <211> 27 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of unknown: Porcine gastrin releasing peptide <400> 56

    Ala Pro Val Ser Val Gly Gly Gly Thr val Leu Ala Lys Met Tyr Pro 15 10 15

    Arg Gly Asn His Trp Ala val Gly His Leu Met 20 25 <210> 57 <211> 27 <212> PRT <213> Homo sapiens <400> 57

    Val Pro Leu Pro Ala Gly Gly Gly Thr val Leu Thr Lys Met Tyr pro 15 10 15

    Arg Gly Asn His Trp Ala val Gly His Leu Met 20 25 <210> 58 <211> 5 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of unknown: Enterokinase peptide <400> 58

    ASP ASp Asp Asp LyS

    1 5 <210> 59 <211> 4 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of unknown: Factor Xa peptide <400> 59 ile Glu Gly Arg <210> 60

    Page 143

    P31003au-pct.txt

    2015268754 16 Dec 2015 <211> 4 <212> PRT <213> Unknown <22O>

    <221> source <223> /note=Description of unknown: Factor Xa peptide <400> 60

    Ile Asp Gly Arg <210> 61 <211> 7 <212> PRT <213> Tobacco etch vi rus <400> 61

    Glu Asn Leu Tyr Phe Gln Gly 1 5 <210> 62 <211> 6 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Thrombin peptide <400> 62

    Leu val Pro Arg Gly Ser

    1 5 <210> 63 <211> 8 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic peptide ) <400> 63

    Leu Glu Val Leu Phe Gln Gly Pro

    1 5 <210> 64 <211> 6 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial sequence: synthetic 6xHis tag <400> 64

    His His His His His His

    1 5 <210> 65

    Page 144

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <211> 4 <212> PRT <213> Unknown <22O>

    <221> source <223> /note=Description of Unknown: Caspase 3 peptide <400> 65

    Asp Met Gin Asp <210> 66 <211> 6 <212> PRT <213> unknown <220>

    <221> source <223> /note=Description of Unknown: Thrombin peptide <400> 66

    Leu Val Pro Arg Gly Ser

    1 5 <210> 67 <211> 10 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: ADAM17 peptide <400> 67

    Pro Leu Ala Gin Ala val Arg Ser Ser Ser

    1 5 10 <210> 68 <211> 10 <212> PRT <213> Homo sapiens <400> 68

    Ser Lys Gly Arg Ser Leu Ile Gly Arg Val 1 5 10 <210> 69 <211> 6 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Elastase peptide <400> 69

    Met Glu Ala val Thr Tyr

    1 5 <210> 70 <211> 4 <212> PRT

    Page 145

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Granzyme peptide <400> 70 ile Glu Pro Asp <210> 71 <211> 4 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Caspase 2 peptide <400> 71

    Asp Val Ala Asp <210> 72 <211> 4 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Caspase 4 peptide <400> 72

    Leu Glu val Asp <210> 73 <211> 4 <212> PRT <213> unknown <22O>

    <221> source <223> /note=Description of Unknown: Caspase 7 peptide <400> 73

    Asp Glu Val Asp <210> 74 <211> 4 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: caspase 9 peptide <400> 74

    Leu Glu His Asp <210> 75

    Page 146

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <211> 4 <212> PRT <213> unknown <220>

    <221> source <223> /note=Description of unknown: caspase 10 peptide <400> 75

    Ile Glu His Asp <210> 76 <211> 10 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic peptide <220>

    <221> MOD_RES <222> (10)..(10) <223> Cys-methyl ester <400> 76

    Cys His ser Gly Tyr Val Gly Ala Arg Cys 15 10 <210> 77 <211> 12 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Epidermal growth factor peptide <400> 77

    Tyr His Trp Tyr Gly Tyr Thr Pro Gin Asn Val lie

    1 5 10 <210> 78 <211> 116 <212> PRT <213> Homo sapiens <400> 78

    Met Leu Ser cys Arg Leu Gin Cys Ala Leu Ala Ala Leu ser lie val 1 5 10 15 Leu Ala Leu Gly Cys Val Thr Gly Ala Pro Ser Asp Pro Arg Leu Arg 20 25 30 Gin Phe Leu Gin Lys Ser Leu Ala Ala Ala Ala Gly Lys Gin Glu Leu 35 40 45 Ala Lys Tyr Phe Leu Ala Glu Leu Leu Ser Glu Pro Asn Gin Thr Glu

    Page 147

    P31I D03ai j-pc· t.tx· t 50 55 60 Asn ASp Ala Leu Glu Pro Glu Asp Leu Ser Gin Ala Ala Glu Gin Asp 65 70 75 80 Glu Met Arg Leu Glu Leu Gin Arg Ser Ala Asn ser Asn Pro Ala Met 85 90 95 Ala Pro Arg Glu Arg Lys Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr 100 105 110 Phe Thr Ser Cys

    115

    2015268754 16 Dec 2015 <210> 79 <211> 155 <212> PRT <213> Homo sapiens <400> 79

    Met Tyr 1 Arg His Lys 5 Asn Ser Trp Arg Leu Gly Leu Lys Tyr Pro Pro 10 15 Ser Ser Lys Glu Glu Thr Gin Val pro Lys Thr Leu lie Ser Gly Leu 20 25 30 Pro Gly Arg Lys ser Ser Ser Arg val Gly Glu Lys Leu Gin Ser Ala 35 40 45 His Lys Met Pro Leu Ser Pro Gly Leu Leu Leu Leu Leu Leu Ser Gly 50 55 60 Ala Thr Ala Thr Ala Ala Leu Pro Leu Glu Gl y Gl y Pro Thr Gly Arg 65 70 75 80 Asp Ser Glu His Met Gin Glu Ala Ala Gly lie Arg Lys Ser Ser Leu 85 90 95 Leu Thr Phe Leu Ala Trp Trp Phe Glu Trp Thr Ser Gin Ala Ser Al a 100 105 110 Gly Pro Leu Ile Gly Glu Glu Ala Arg Glu val Ala Arg Arg Gin Glu 115 120 125 Gly Ala pro Pro Gin Gin Ser Ala Arg Arg Asp Arg Met Pro Cys Arg 130 135 140 Asn Phe Phe Trp Lys Thr Phe ser Ser Cys Lys 145 150 155

    <210> 80 <211> 7 <212> PRT

    Page 148

    P31003au-pct.txt

    2015268754 16 Dec 2015 <213> Homo sapiens <400> 80

    Asn Phe Phe Trp Lys Thr Phe 1 5 <210> 81 <211> 8 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (1)..(1) <223> /replace=Lys <220>

    <221> misc_feature <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position¹' <400> 81

    Arg Asn Phe Phe Trp Lys Thr Phe

    1 5 <210> 82 <211> 9 <212> PRT <213> Homo sapiens <22O>

    <221> VARIANT <222> (2)..(2) <223> /replace=Lys <220>

    <221> misc_feature <222> (2)..(2) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position ' <400> 82

    Cys Arg Asn Phe Phe Trp Lys Thr Phe

    1 5 <210> 83 <211> 10 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (1)..(1) <223> /replace=Gly <220>

    <221> misc_feature <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said

    Page 149

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 position <220>

    <221> VARIANT <222> (3)..(3) <223> /replace=Lys <220>

    <221> misc_feature <222> (3)..(3) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position ' <400> 83

    Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe

    15 10 <210> 84 <211> 8 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (8)..(8) <223> /replace=Thr <220>

    <221> misc_feature <222> (8)..(8) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <400> 84

    Asn Phe Phe Trp Lys Thr Phe Ser

    1 5 <210> 85 <211> 9 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (8)..(8) <223> /replace=Thr <220>

    <221> misc_feature <222> (8)..(8) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <400> 85

    Asn Phe Phe Trp Lys Thr Phe ser Ser

    1 5 <210> 86 <211> 10 <212> PRT

    Page 150

    P31OO3au-pct.txt <213> Homo sapiens

    2015268754 16 Dec 2015 <220>

    <221> VARIANT <222> (8)..(8) <223> /rep1ace=Thr <220>

    <221> misc_feature <222> (8)..(8) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position ' <400> 86

    Asn Phe Phe Trp Lys Thr Phe Ser Ser cys

    15 10 <210> 87 <211> 9 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (1)..(1) <223> /replace=Lys <220>

    <221> misc_feature <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position¹' <220>

    <221> VARIANT <222> (9)..(9) <223> /replace=Thr <220>

    <221> misc_feature <222> (9)..(9) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position ' <400> 87

    Arg Asn Phe Phe Trp Lys Thr Phe Ser

    1 5 <210> 88 <211> 10 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (1)..(1) <223> /replace=Lys <220>

    <221> misc_feature page 151

    P31003au-pct. txt

    2015268754 16 Dec 2015 <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <220>

    <221> VARIANT <222> (9)..(9) <223> /replace=Thr <220>

    <221> misc_feature <222> (9)..(9) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position' <400> 88

    Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser

    1 5 10 <210> 89 <211> 11 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (1)..(1) <223> /replace©'Lys <220>

    <221> misc_feature <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position' <22O>

    <221> VARIANT <222> (9)..(9) <223> /replace=Thr <220>

    <221> misc_feature <222> (9)..(9) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <400> 89

    Arg Asn Phe phe Trp Lys Thr Phe ser ser cys

    1 5 10 <210> 90 <211> 10 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (2)..(2) <223> /replace=Lys

    Page 152

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <22Ο>

    <221> misc_feature <222> ¢2)..(2) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <220>

    <221> VARIANT <222> (10)..(10) <223> /replace=Thr <220>

    <221> misc_feature <222> (10)..(10) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <400> 90

    Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser

    1 5 10 <210> 91 <211> 11 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (2)..(2) <223> /rep1ace=”Lys <22O>

    <221> misc_feature <222> (2)..(2) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position¹' <220>

    <221> VARIANT <222> (10)..(10) <223> /replace=Thr <220>

    <221> misc_feature <222> (10)..(10) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position¹' <400> 91

    Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser

    15 10 <210> 92 <211> 12 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (2)..(2)

    Page 153

    P31003au-pct. txt

    2015268754 16 Dec 2015 <223> /replace=Lys <220>

    <221> misc_feature <222> (2)..(2) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position ' <220>

    <221> VARIANT <222> (10)..(10) <223> /replace=Thr <220>

    <221> misc_feature <222> (10)..(10) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <400> 92

    Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser cys

    1 5 10 <210> 93 <211> 11 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (1)..(1) <223> /replace=Gly <220>

    <221> misc_feature <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position ' <220>

    <221> VARIANT <222> (3)..(3) <223> /replace=Lys <220>

    <221> misc_feature <222> (3)..(3) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <220>

    <221> VARIANT <222> (11)..(11) <223> /replace=Thr <220>

    <221> misc_feature <222> (11)..(11) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position¹'

    Page 154

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <400> 93

    Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser 1 5 10 <210> 94 <211> 12 <212> PRT <213> Homo sapiens <22O>

    <221> VARIANT <222> (1)..(1) <223> /replace=Gly <220>

    <221> misc_feature <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position'¹ <220>

    <221> VARIANT <222> (3)..(3) <223> /replace=Lys <220>

    <221> misc_feature <222> (3)..(3) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <220>

    <221> VARIANT <222> (11)..(11) <223> /replace=Thr <220>

    <221> misc_feature <222> (11)..(11) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position ¹ <400> 94

    Pro cys Arg Asn Phe Phe Trp Lys Thr Phe ser ser

    1 5 10 <210> 95 <211> 12 <212> PRT <213> Homo sapiens <220>

    <221> VARIANT <222> (1)..(1) <223> /replace=Gly <220>

    <221> misc_feature <222> (1)..(1) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said

    Page 155

    P31003au-pct.txt position

    2015268754 16 Dec 2015 <220>

    <221> VARIANT <222> (3)..(3) <223> /replace=Lys <220>

    <221> misc_feature <222> (3)..(3) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <22O>

    <221> VARIANT <222> (11)..(11) <223> /replace=Thr” <220>

    <221> misc_feature <222> (11)..(11) <223> /note=Residue given in the sequence has no preference with respect to the residue in the annotation for said position <400> 95

    Pro Cys Arg Asn Phe Phe Trp Lys Thr phe Ser Cys

    15 10 <210> 96 <211> 28 <212> PRT <213> Homo sapiens <400> 96 ser Ala Asn Ser Asn Pro Ala Met Ala Pro Arg Glu Arg Lys Ala Gly 15 10 15

    Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr ser Cys 20 25 <210> 97 <211> 14 <212> PRT <213> Homo sapiens <400> 97

    Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys 1 5 10 <210> 98 <211> 29 <212> PRT <213> Homo sapiens <400> 98

    Gln Glu Gly Ala Pro Pro Gln Gln ser Ala Arg Arg Asp Arg Met Pro 15 10 15

    Cys Arg Asn Phe Phe Trp Lys Thr Phe ser Ser Cys Lys 20 25

    Page 156

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <210> 99 <211> 29 <212> PRT <213> Homo sapiens <400> 99

    Gin Glu Arg Pro Pro Leu Gin Gin Pro Pro His Arg Asp Lys Lys pro 15 10 15

    Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 20 25 <210> 100 <211> 29 <212> PRT <213> Homo sapiens <400> 100

    Gin Glu Arg Pro Pro Pro Gin Gin Pro Pro His Leu Asp Lys Lys Pro 15 10 15

    Cys Lys Asn Phe Phe Trp Lys Thr Phe Ser ser Cys Lys 20 25 <210> 101 <211> 17 <212> PRT <213> Homo sapiens <400> 101

    Asp Arg Met Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys 15 10 15

    Lys <210> 102 <211> 14 <212> PRT <213> Homo sapiens <400> 102

    Pro Cys Arg Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 1 5 10 <210> 103 <211> 14 <212> PRT <213> Homo sapiens <400> 103

    Pro cys Lys Asn Phe Phe Trp Lys Thr Phe Ser Ser Cys Lys 15 10 <210> 104 <211> 10 <212> PRT <213> Unknown

    Page 157

    P31OO3au-pct.txt

    2015268754 16 Dec 2015 <220>

    <221> source <223> /note=Description of unknown: Bombesin peptide <400> 104

    Gly Asn Gin Trp Ala val Gly His Leu Met 1 5 10 <210> 105 <211> 10 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Neuromedin B peptide <400> 105

    Gly Asn Leu Trp Ala Thr Gly His Phe Met

    1 5 10 <210> 106 <211> 10 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of unknown: Neuromedin C peptide <400> 106

    Gly Asn His Trp Ala val Gly His Leu Met

    15 10 <210> 107 <211> 9 <212> PRT <213> Unknown <220>

    <221> source <223> /note=Description of Unknown: Litorin peptide <220>

    <221> MOD_RES <222> ¢1)..(1) <223> pyro-Glutamate <400> 107

    Glu Gin Trp Ala Val Gly His Phe Met 1 5 <210> 108 <211> 10 <212> PRT <213> Homo sapiens <400> 108

    Gly Asn His Trp Ala val Gly His Leu Met

    Page 158

    P31OO3au-pct.txt

    1 5 10

    2015268754 16 Dec 2015 <210> 109 <211> 23 <212> PRT <213> influenza virus <400> 109

    Gly Leu Phe Gly Ala lie Ala Gly Phe lie Glu Asn Gly Trp Glu Gly 15 10 15

    Met lie Asp Gly Trp Tyr Gly 20 <210> 110 <211> 23 <212> PRT <213> Artificial sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic peptide <400> 110

    Val Arg Gly ile lie Pro Phe Lys Thr Lys Ser Leu Asp Glu Gly Tyr 15 10 15

    Asn Lys Ala Leu Asn Asp Leu 20 <210> 111 <211> 23 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: Synthetic peptide <400> 111 val Asp Gly Ile ile Thr ser Lys Thr Lys Ser Leu lie Glu Gly Arg 15 10 15

    Asn Lys Ala Leu Asn Leu Gln 20 <210> 112 <211> 23 <212> PRT <213> Artificial Sequence <220>

    <221> source <223> /note=Description of Artificial Sequence: synthetic peptide <400> 112

    Val Asp Gly Ile Ile Thr ser Lys Thr Lys Ser Asp Asp Asp Asp Lys 15 10 15

    Page 159

    P31OO3au-pct.txt

    2015268754 16 Dec 2015

    Asn Lys Ala Leu Asn Leu Gin 20

    Page 160