NZ709739B2 - Anti-il-17a antibodies and their use in treating autoimmune and inflammatory disorders - Google Patents

Abstract

The present disclosure relates to antibodies and proteins comprising an antigen-binding portion thereof that specifically bind to the pro-inflammatory cytokine IL-17A. The disclosure more specifically relates to specific antibodies and proteins that are IL-17A antagonists (inhibit the activities of IL-17A and IL-17AF), are capable of inhibiting IL- 17A induced cytokine production in in vitro assays, having an inhibitory effect in an antigen-induced arthritis model in vivo, and can selectively bind to homodimeric IL-17A and heterodimeric IL-17AF across several species. The disclosure further relates to compositions and methods of use for said antibodies and proteins to treat pathological disorders that can be treated by inhibiting IL-17A or IL17AF mediated activity, such as rheumatoid arthritis, psoriasis, systemic lupus erythematosus (SLE), lupus nephritis, chronic obstructive pulmonary disease, asthma or cystic fibrosis or other autoimmune and inflammatory disorders. IL-17A and IL-17AF), are capable of inhibiting IL- 17A induced cytokine production in in vitro assays, having an inhibitory effect in an antigen-induced arthritis model in vivo, and can selectively bind to homodimeric IL-17A and heterodimeric IL-17AF across several species. The disclosure further relates to compositions and methods of use for said antibodies and proteins to treat pathological disorders that can be treated by inhibiting IL-17A or IL17AF mediated activity, such as rheumatoid arthritis, psoriasis, systemic lupus erythematosus (SLE), lupus nephritis, chronic obstructive pulmonary disease, asthma or cystic fibrosis or other autoimmune and inflammatory disorders.

Description

ANTI-IL-17A ANTIBODIES AND THEIR USE IN TREATING AUTOIMMUNE AND INFLAMMATORY DISORDERS RELATED APPLICATIONS The present disclosure claims priority to US 406, filed 8 February 2013, which is incorporated by reference herein in its ty.

FIELD OF THE INVENTION The present disclosure relates to antibodies and proteins comprising an antigen-binding portion thereof that specifically bind to IL-17A. The disclosure more specifically relates ’IO to specific dies and ns that inhibit the effects of IL-17A and are capable of inhibiting IL-17A-induced activity, as well as compositions and methods of use for said antibodies and proteins, e.g. to treat pathological ers that can be treated by inhibition of IL-17A signaling, for example mune and inflammatory disorders such as rheumatoid arthritis, psoriasis, systemic lupus erythematosus (SLE), lupus nephritis, multiple sclerosis, or chronic obstructive pulmonary disease, asthma or cystic fibrosis.

BACKGROUND OF THE INVENTION Interleukin-17A A also sometimes called IL-17) is the central lymphokine produced by a newly defined subset of inflammatory T cells, the Th1? cells. In several animal models, these cells are pivotal for various autoimmune and inflammatory processes. Increased levels of IL-17A have been associated with uveitis (Ambadi-Obi, et al 2007, Nature Med; 13:711-718), rheumatoid arthritis (RA), psoriasis, airway inflammation, chronic ctive pulmonary disease , inflammatory bowel disease (Crohn’s disease and ulcerative colitis), allograft rejection, cancer, intra- peritoneal abscesses and adhesions, and multiple sclerosis (Weaver, et al 2007, Annu Rev Immunol; 25:821-852; Witowski et al 2004, Cell Mol Life Sci; -579). Th1? cells can rapidly initiate an inflammatory response that is dominated by neutrophils ec, et al 2009, NEJM; 361:888—98). lL-17A was ally identified as a transcript from a rodent T-cell hybridoma. It is the founding member of a group of cytokines called the lL-17 family. Known as CTLA8 in rodents, lL-17A shows high homology to viral lL-17A encoded by an open reading frame of the T-lymphotropic ovirus herpesvirus saimiri (Rouvier E, et al 1993, J. lmmunol. 150: 5445—56). lL-17A is a cytokine that acts as a potent mediator in d-type reactions by increasing chemokine production in various tissues to recruit monocytes and neutrophils to the site of inflammation, similar to eron gamma. The lL-17 family functions in the role of proinflammatory cytokines that respond to the invasion of the immune system by 1O extracellular pathogens and induces destruction of the pathogen’s cellular matrix. lL- 17A acts synergistically with tumor necrosis factor and eukin-1 (Miossec P, et al 2009, N. Engl. J. Med. 361:888—98).

To elicit its functions, lL-17A binds to a type | cell surface receptor called lL-17R of which there are at least two variants, lL-17RA and lL-17RC (Pappu R, et al 2012, Trends lmmunol.; 33:343-9). lL-17RA binds lL-17A, lL-17AF and lL-17F and is expressed in multiple tissues: vascular endothelial cells, peripheral T cells, B cell lineages, fibroblast, lung, myelomonocytic cells, and marrow stromal cells (Kolls JK, Linden A 2004, lmmunity 21:467—76; Kawaguchi M, et al 2004, J. Allergy Clin. lmmunol. 114:1265—73; y TA, et al 2003, Cytokine Growth Factor Rev. 14:155—74). 2O In addition to lL-17A, members of the lL-17 family include lL-17B, lL-17C, lL-17D, lL- 17E (also called lL-25), and lL-17F. All members of the lL-17 family have a r protein ure, with four highly conserved cysteine es critical to their 3- dimensional shape. Phylogenetic is reveals that among lL-17 family members, the lL-17F isoforms 1 and 2 (ML-1) have the highest homology to lL-17A (sharing 55 and 40% amino acid identity to lL-17A respectively), followed by lL-17B (29%), lL-17D (25%), lL-17C (23%), and lL-17E being most distantly related to lL-17A (17%). These cytokines are all well conserved in mammals, with as much as 62—88% of amino acids conserved between the human and mouse homologs (Kolls JK, Linden A 2004, Immunity 21 :467—76). lL-17A is a 155-amino acid protein that is a disulfide-linked, homodimeric, ed glycoprotein with a molecular mass of 35 kDa (Kolls JK, Linden A 2004, lmmunity 21 :467—76). The structure of lL-17A consists of a signal e ed by the amino acid region characteristic of the lL-17 family. An ed glycosylation site on the protein was first identified after purification of the n revealed two bands in standard SDS-PAGE analysis, one at 15 kDa and another at 20 kDa. Comparison of different s of the lL-17 family revealed four conserved cysteines that form two ide bonds (Yao Z, et al 1995, J. lmmunol. 155:5483—6). lL-17 is unique in that it bears no resemblance to other known interleukins. Furthermore, lL-17 bears no resemblance to any other known proteins or structural domains (Kolls JK, Linden A2004, lmmunity 21:467—76). Generally, other members of the lL-17 family such as lL-17F form 1O homodimers (like lL-17A). lL-17A is also known to form a heterodimer with lL-17F under certain circumstances.

Heterodimeric lL-17AF is also produced by Th1? cells following stimulation by lL-23. lL-17AF is thought to signal h the lL-17RA and lL-17RC receptors like lL-17A and lL-17F. The biological functions of lL-17AF are similar to those of lL-17A and lL-17F.

Stimulation of target cells by lL-17AF induces the production of a variety of chemokines, in addition to ain/vay neutrophilia in appropriate circumstances. lL-17AF is considered to be less potent in these activities than homodimeric lL-17A, but more potent than homodimeric lL-17F. For example, if the potency of lL-17A is 1, then the relative potency of lL-17AF is about 1/10 of that of lL-17A and the relative y of lL-17F is 2O about 1/100 of that of lL-17A. Human and mouse lL-17AF both show activity on mouse cells. lL-17AF consists of a total of 271 amino acids and has a molecular weight of approximately 30.7kDa (data from product description of Human lL-17AF Heterodimer from Shenandoah Biotechnology).

A number of relevant crystal structures have been published. These e the crystal structure for homodimeric lL-17F (Hymowitz et al 2001, EMBO J, 19:5332—5341).

The crystal structure of lL-17F in complex with the receptor lL-17RA has also been published (Ely et al., 2009 Nature Immunology 10:1245—1251). In on at least one crystal ure of lL-17A in complex with the Fab fragment of an dy has been published (Gerhardt et al., 2009 Journal of lar Biology, 5:905-921).

W0 2014/122613 Several inflammatory and autoimmune diseases including psoriasis are linked to exacerbated Th1 and/or Th17 ses. Many of them are currently treated either with l immunosuppressants or very selectively acting biologicals such as anti-TNF-d antibodies that are not effective in all patients. These were found to increase the risk for infections and to become ineffective after repeated treatment. Therefore, there is an unmet medical need for treatments with sed safety profiles and simultaneous capacity to induce long-term remission or cure of the disease.

Numerous immune regulatory functions have been reported for the lL-17 family of cytokines, it is presumed due to their induction of many immune ing molecules. 1O The most notable role of lL-17A is its involvement in inducing and ing proinflammatory responses. lL-17A is also associated with allergic responses. lL-17 induces the production of many other cytokines (such as lL-6, G-CSF, , lL-1B, TGF-B, TNF-d), chemokines ding lL-8, GRO-d, and , and prostaglandins (e.g., PGE2) from many cell types (fibroblasts, endothelial cells, epithelial cells, keratinocytes, and macrophages). The release of nes causes many functions, such as ain/vay remodeling, a characteristic of lL-17A ses. The increased expression of chemokines attracts other cells including neutrophils but not eosinophils. lL-17 on is also essential to a subset of CD4+ T-cells called T helper 17 (Th1?) cells. As a result of these roles, the lL-17 family has been linked to many immune/autoimmune related diseases including rheumatoid arthritis, asthma, lupus, allograft rejection and anti-tumor immunity (Aggarvval S, Gurney AL 2002, J. Leukoc.

Biol. 71:1—8). Additionally, links have been drawn to further conditions such as osteoarthritis, septicemia, septic or endotoxic shock, allergic reactions, bone loss, psoriasis, ischemia, systemic sclerosis, fibrosis, and stroke.

Thus, there is a need for ic dies that antagonize the effects of lL-17A and are capable of inhibiting lL-17A induced activity, and especially compositions and methods of use for said antibodies to treat pathological disorders that can be treated by inhibition of lL-17A signaling.

SUMMARY OF THE INVENTION Therefore, in one aspect, the disclosure es an isolated antibody or protein comprising an antigen-binding portion of an antibody, comprising CDR amino acid sequences having at least 95% identity to those encoded by SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3, and CDR amino acid sequence having at least 64% identity to those d by SEQ ID NO: 42, SEQ ID NO: 23 and SEQ ID NO: 11, and n said antibody or molecule specifically binds to homodimeric IL-17A and heterodimeric IL-17AF, but does not specifically bind to homodimeric IL-17F.

In a ular aspect the invention provides an isolated antibody or protein comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein: a) said VH comprises, in sequence, the three complementarity determining regions (CDRs) set forth in SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3, and b) said VL comprises, in sequence, the three CDRs set forth in SEQ ID NO: 42, SEQ ID NO: 23 and SEQ ID NO: 11, and wherein said antibody or protein specifically binds to homodimeric IL-17A and heterodimeric IL-17AF, but does not specifically bind to homodimeric IL-17F.In one embodiment, the IL-17A, IL-17AF or IL-17F are selected from one or more, such as two or three or more, of cynomolgus , rhesus macaque monkey, marmoset monkey, rat, mouse or human. In one specific embodiment, the IL-17A, IL-17AF or IL-17F is from human.

In one specific embodiment, the IL-17A, IL-17AF or IL-17F is from human and mouse. In one specific embodiment, the IL-17A, IL-17AF or IL-17F is from cynomolgus monkey, rhesus macaque monkey, marmoset monkey, rat, mouse and human.

In one specific ment, the isolated antibody or protein comprising an antigen-binding portion thereof of the disclosure, comprises an amino acid ce having at least 95% identity to SEQ ID NO: 12, and an amino acid sequence having at least 90% identity to SEQ ID NO: 43. In one embodiment, the ed antibody or comprises an amino acid sequence having at least 95% identity to SEQ ID NO: 14, and an amino acid sequence having at least 95% identity to SEQ ID NO: 44.

In one embodiment, the isolated antibody or protein comprising an antigen-binding n thereof comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 domain ed from the group consisting of a) a light chain CDR1 domain of SEQ ID NO: 73, wherein the first variable amino acid is selected from the group consisting of Gly (G) and Val (V); the second variable amino acid is ed from the group consisting of Tyr (Y), Asn (N) and Ile (I); the third variable amino acid is selected from the group consisting of Trp (W) and Ser (S); and the fourth le amino acid is selected from the group consisting of Glu (E) and Ala (A); b) a light chain CDR2 domain of SEQ ID NO: 74, wherein the variable amino acid is selected from the group consisting of Asn (N) and Gln (Q); and c) a light chain CDR3 domain of SEQ ID NO: 75, wherein the variable amino acid is selected from the group consisting of Asn (N) and Asp (D).

In one embodiment, the isolated antibody or protein sing an antigen-binding portion thereof comprises heavy chain CDRs comprising, in ce, a) SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3, and light chain CDRs comprising, in sequence, b) SEQ ID NO: 42, SEQ ID NO: 23 and SEQ ID NO: 11, c) SEQ ID NO: 42, SEQ ID NO: and SEQ ID NO: 11, d) SEQ ID NO: 34, SEQ ID NO: 23 and SEQ ID NO: 11, e) SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24, or f) SEQ ID NO: 9, SEQ ID NO: and SEQ ID NO:11.

In one specific embodiment, the isolated antibody or protein comprising an antigen- binding portion thereof comprises heavy chain CDRs, in sequence, SEQ ID NO: 7, SEQ 1O ID NO: 8 and SEQ ID NO: 3 and light chain CDRs, in sequence, SEQ ID NO: 42, SEQ ID NO: 23 and SEQ ID NO:11.

In another specific embodiment, the isolated antibody or protein comprising an antigen- binding n thereof comprises heavy chain CDRs, in sequence, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3 and light chain CDRs, in sequence SEQ ID NO: 42, SEQ ID N010 and SEQ ID NO:11.

In another specific embodiment, the isolated dy or protein sing an antigen- binding portion thereof comprises heavy chain CDRs, in sequence, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3 and light chain CDRs, in sequence, SEQ ID NO: 34, SEQ ID NO: 23 and SEQ ID NO:11.

In another specific ment, the isolated antibody or protein comprising an antigen- binding portion thereof comprises heavy chain CDRs, in sequence, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3 and light chain CDRs, in sequence, SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24.

In another specific embodiment, the isolated antibody or protein comprising an antigen- binding portion thereof comprises heavy chain CDRs, in sequence, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3 and light chain CDRs, in ce, SEQ ID NO: 9, SEQ ID N010 and SEQ ID NO:11.

In one ment, the isolated dy or protein comprising an antigen-binding portion thereof comprises an immunoglobulin heavy chain comprising a) SEQ ID NO: 12, and an immunoglobulin light chain comprising b) SEQ ID NO: 43, c) SEQ ID NO: 53, d) SEQ ID NO: 35, e) SEQ ID NO: 25, or f) SEQ ID NO: 13.

In one specific embodiment, the isolated antibody or n sing an antigenbinding portion thereof comprises an immunoglobulin heavy chain according to SEQ ID NO: 12 and an immunoglobulin light chain according to SEQ ID NO: 43.

In one specific embodiment, the isolated antibody or protein comprising an antigen- binding portion thereof ses an immunoglobulin heavy chain according to SEQ ID NO: 12 and an immunoglobulin light chain ing to SEQ ID NO: 53.

In one specific embodiment, the ed antibody or protein comprising an antigen- 1O binding portion thereof comprises an immunoglobulin heavy chain according to SEQ ID NO: 12 and an globulin light chain according to SEQ ID NO: 35.

In one specific embodiment, the isolated antibody or protein comprising an antigen- binding portion thereof comprises an immunoglobulin heavy chain according to SEQ ID NO: 12 and an immunoglobulin light chain according to SEQ ID NO: 25.

In one specific embodiment, the isolated antibody or protein comprising an n- binding portion thereof comprises an immunoglobulin heavy chain according to SEQ ID NO: 12 and an immunoglobulin light chain according to SEQ ID NO: 13.

In one embodiment, the isolated antibody or protein comprising an antigen-binding portion thereof comprises an immunoglobulin heavy chain sing a) SEQ ID NO: 14, and an immunoglobulin light chain comprising b) SEQ ID NO: 44, c) SEQ ID NO: 54, d) SEQ ID NO: 36, e) SEQ ID NO: 26, or f) SEQ ID NO: 15.

In one ic embodiment, the isolated antibody or protein comprising an antigen- binding portion thereof comprises an immunoglobulin heavy chain according to SEQ ID NO: 14, and an immunoglobulin light chain according to SEQ ID NO: 44.

In one specific embodiment, the isolated antibody or protein comprising an antigen- binding n thereof comprises an immunoglobulin heavy chain according to SEQ ID NO: 14, and an immunoglobulin light chain according to SEQ ID NO: 54.

In one specific embodiment, the isolated dy or protein comprising an antigen- binding portion thereof comprises an globulin heavy chain according to SEQ ID NO: 14, and an immunoglobulin light chain according to SEQ ID NO: 36.

In one specific embodiment, the isolated antibody or protein comprising an antigenbinding portion thereof comprises an globulin heavy chain according to SEQ ID NO: 14, and an immunoglobulin light chain according to SEQ ID NO: 26.

In one specific embodiment, the isolated antibody or protein comprising an antigenbinding portion thereof ses an immunoglobulin heavy chain according to SEQ ID NO: 14, and an immunoglobulin light chain according to SEQ ID NO: 15.

In one aspect of the disclosure, an isolated antibody or n comprising an antigen- binding portion f is provided, which binds to the same epitope as an isolated antibody or molecule according to specific embodiments of the disclosure.

In one embodiment, the isolated dy or n comprising an antigen-binding portion thereof binds to an IL-17A epitope, such as a human IL-17A epitope, which comprises Arg78, Glu80, and Trp90.

The IL-17A epitope may further comprise Tyr85 or Arg124.

In one embodiment, the IL-17A epitope, such as a human IL-17A epitope, further comprises one or more of Pro82, Ser87 or Val88.

In one aspect of the disclosure, an isolated antibody or protein comprising an antigen- binding portion thereof is provided, which comprises an antigen recognition surface having epitope recognition characteristics equivalent to an antibody or molecule according to ic embodiments.

In one aspect of the disclosure, an isolated antibody or protein comprising an antigen- binding n thereof is provided which is cross-blocked from binding to IL-17A, such as human IL-17A, or IL-17AF, such as human F, by at least one antibody or n comprising an antigen-binding portion thereof according to ic embodiments.

In one embodiment, the antibody or n comprising an antigen-binding portion thereof does not specifically bind to a) any one or more of human lL-17F homodimer, IL- 178 homodimer, lL-17C homodimer, lL-17D homodimer, lL-17E homodimer, and/or b) any one or more of cynomolgus monkey lL-17F homodimer, mouse lL-17F homodimer, and/or c) any one or more of other human cytokines selected from the group consisting of lL-1, lL-3, lL-4, lL-6, lL-8, glFN, TNF alpha, EGF, GMCSF, TGF beta 2, and/or d) any one or more of other mouse cytokines, selected from the group consisting of lL-1 b, lL-2, lL-4, lL-6, lL-12, IL18, lL23, lFN or TNF.

In one embodiment, the antibody or protein comprising an antigen-binding portion 1O thereof binds to lL-17A, such as human lL-17A, so that the antibody or protein comprising an antigen-binding portion thereof inhibits or blocks binding between lL-17A and its receptor, and reduces or neutralizes lL-17A activity.

In one embodiment, the binding affinity of the antibody or protein comprising an antigen- binding portion thereof for human lL-17A is 100 nM or less, 10 nM or less, 1 nM or less, 100 pM or less, or 10 pM or less as measured by BiacoreTM. In a specific embodiment, the binding affinity of the antibody or n comprising an antigen-binding portion thereof for human lL-17A is below 200 pM, or below 100 pM, as measured by B iacoreTM.

In one embodiment, the antibody or n sing an antigen-binding portion thereof is capable of inhibiting lL-6 secretion, or GRO-alpha secretion when assessed in vitro, preferably using cultured chondrocytes or lasts.

In one embodiment, the antibody or protein sing an antigen-binding n f is capable of inhibiting knee swelling in an antigen induced arthritis experimental model in vivo, such as a rat del.

In one ment, the antibody or protein sing an antigen-binding portion thereof is conjugated to a r active moiety.

The antibody or protein comprising an antigen-binding portion thereof may be monoclonal antibody or an antigen-binding portion thereof, preferably a chimeric, humanized, or human antibody or portion thereof.

W0 2014/122613 In an aspect of the disclosure, a pharmaceutical composition is provided, sing an antibody or protein sing an antigen-binding portion thereof according to embodiments of the sure, in combination with one or more pharmaceutically acceptable excipient, diluent or carrier.

In an embodiment, the pharmaceutical composition comprises one or more additional active ients.

In one specific embodiment, said pharmaceutical composition is a lyophilisate. In another specific ment, the pharmaceutical composition is a liquid formulation comprising a therapeutically acceptable amount of an antibody or molecule of the 1O disclosure, preferably prepared as a pre-filled syringe.

The sure further relates to the use of said antibody or protein comprising an antigen-binding n thereof of the disclosure, in particular XABl, XABZ, XABS, XAB4 or XAB5 antibodies, for use as a ment, more preferably, for the treatment of a pathological disorder that is mediated by lL-17A or that can be treated by inhibition of lL-17A signaling, or lL-6 or GRO-alpha secretion.

In one specific embodiment, the antibodies or proteins comprising an antigen-binding portion thereof of the disclosure may be used for the treatment of mune and inflammatory disorders, such as tis, rheumatoid arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, asthma, multiple sclerosis, or cystic fibrosis.

In one aspect of the sure, a use of said antibody or protein comprising an antigen- binding portion thereof of the disclosure, in particular XABl, XABZ, XABS, XAB4 or XAB5 antibodies, in the manufacture of a medicament for use in the ent of a pathological disorder mediated by lL-17A or that can be treated by inhibiting lL-6 or pha secretion is provided.

In one specific embodiment, the a pathological disorder mediated by lL-17A or that can be treated by inhibiting lL-6 or GRO-alpha secretion is an inflammatory disorder or condition, such as arthritis, toid arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, asthma, multiple sclerosis or cystic fibrosis.

W0 2014/122613 In one aspect of the disclosure, a method of treating a pathological disorder mediated by lL-17A, or that can be treated by inhibiting lL-6 or GRO-alpha secretion is ed, said method comprising stering an effective amount of an isolated antibody or molecule ing to the disclosure, in particular XAB1, XAB2, XAB3, XAB4 or XAB5 antibodies, such that the condition is alleviated.

In an embodiment, the condition is an inflammatory disorder or condition, such as tis, rheumatoid arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, asthma, le sclerosis or cystic fibrosis.

The disclosure also relates to the means for producing the antibodies or protein 1O comprising an antigen-binding portion thereof of the disclosure. Such means e isolated c acid molecules encoding at least the heavy and/or light variable region(s) of the antibody or protein of the disclosure or cloning expression vectors comprising such nucleic acids, in particular, for the inant production of an antibody or protein according to the disclosure, for e XAB1, XAB2, XAB3, XAB4 or XAB5, in a host cell. In a specific embodiment, such cloning or sion vector comprises at least one nucleic acid sequence selected from the group consisting of SEQ ID NO: 18, 31, 51, 19, 28, 32, 38, 40, 46, 48, 52, 56, and 58. In r embodiment, it comprises at least one of the following coding sequences of variable heavy and light chain sequences selected from the group consisting of SEQ ID NO: 16, 29, 49, 17, 27, 30, 37, 39, 45, 47, 50, 55, and 57, operatively linked to suitable promoter sequences, which are well known to a person skilled in the art.

In an ment, the nucleic acid molecule is a messenger RNA (mRNA), The disclosure further s to a host cell comprising one or more cloning or expression vectors as described above and to the process for the production of an antibody or protein comprising an antigen-binding portion thereof of the disclosure, in particular XAB1, XAB2, XAB3, XAB4 or XAB5, said process comprising culturing the host cell, purifying and recovering said antibody or protein.

Definitions In order that the present disclosure may be more readily tood, certain terms are first defined. Additional definitions are set forth throughout the detailed description.

W0 2014/122613 The term "immune response" refers to the action of, for example, lymphocytes, antigen presenting cells, phagocytic cells, granulocytes, and soluble macromolecules produced by the above cells or the liver (including antibodies, cytokines, and complement) that results in selective damage to, ction of, or elimination from the human body of invading pathogens, cells or tissues infected with ens, cancerous cells, or, in cases of munity or pathological inflammation, normal human cells or tissues.

A "signal transduction y" or "signaling ty" refers to a biochemical causal relationship generally initiated by a protein-protein interaction such as binding of a growth factor to a receptor, resulting in transmission of a signal from one portion of a 1O cell to another portion of a cell. In general, the transmission involves specific phosphorylation of one or more tyrosine, serine, or ine residues on one or more proteins in the series of reactions causing signal transduction. Penultimate processes typically include nuclear events, resulting in a change in gene expression.

A naturally ing "antibody" is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds. Each heavy chain is comprised of a heavy chain le region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant . The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity ining regions (CDR), interspersed with regions that are more conserved, termed ork regions (FR).

Each VH and VL is composed of three CDRs and four FRs arranged from amino- terminus to y-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies may mediate the g of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

The terms "complementarity determining region," and "CDR," as used herein refer to the sequences of amino acids within antibody variable regions which confer antigen specificity and binding affinity. In general, there are three CDRs in each heavy chain W0 2014/122613 variable region (HCDR1, HCDR2, HCDR3) and three CDRs in each light chain variable region (LCDR1, LCDR2, LCDR3).

The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of nown schemes, including those described by Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD t" numbering scheme), Al- Lazikani et al., (1997) JMB 273,927-948 hia" numbering scheme). For example, for classic formats, under Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are ed 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 1O (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). Under Chothia the CDR amino acids in the VH are numbered 26-32 (HCDR1’), 52-56 (HCDR2’), and 95- 102 (HCDRS’); and the amino acid residues in VL are numbered 26-32 (LCDR1’), 50-52 (LCDR2’), and 91-96 (LCDR3’). By combining the CDR definitions of both Kabat and Chothia, the CDRs consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL.

The term "antigen-binding portion" of an antibody (or simply "antigen portion"), as used herein, refers to full length or one or more fragments of an antibody, such as a protein, that retain the ability to specifically bind to an n (e.g., a portion of lL-17A). It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding n" of an antibody include a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; a F(ab)2 nt, a bivalent fragment sing two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CH1 domains; a Fv fragment ting of the VL and VH domains of a single arm of an antibody; a dAb fragment (Ward et al. 1989, Nature 341:544-546), which consists of a VH domain; and an isolated complementarity ining region (CDR), or any fusion proteins comprising such antigen-binding n. ingly, the term "antigen-binding portion" may also refer to the portions ponding to the antibody of the disclosure that may be comprised within alternative W0 2014/122613 2014/058854 orks or scaffolds such as camelid antibodies or ‘non-antibody’ les as described below.

Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that s them to be made as a single chain protein in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. 1988, Science 3-426; and Huston et al. 1988, Proc. Natl. Acad. Sci. 85:5879— 5883). Such single chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an dy. These antibody fragments are obtained ’IO using conventional techniques known to those of skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies.

An "isolated antibody", as used herein, refers to an antibody that is substantially free of other antibodies having different nic specificities (e.g., an isolated antibody that specifically binds to IL-17A, such as human IL-17A, is substantially free of antibodies that specifically bind to other antigens than IL-17A). An ed antibody that specifically binds to IL-17A may, however, have cross-reactivity to other ns, such as IL-17A molecules from other species, or IL-17A heterodimers, such as IL-17AF.

Moreover, an isolated antibody may be substantially free of other cellular material and/or chemicals.

The terms lonal antibody" or "monoclonal antibody composition" as used herein refer to a preparation of antibody molecules of single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular e.

The term IL-17A refers to human IL-17A as defined in SEQ ID NO: 76 or SEQ ID NO: 78 unless otherwise described. The term IL-17F refers to human IL-17F as defined in SEQ ID NO: 77 unless othen/vise described. IL-17AF is a heterodimer of an IL-17A subunit and an IL-17F subunit, as will be appreciated by a person skilled in the art.

Recombinant proteins, designated with the prefix "r", from different species were used in the assays described below. For example, recombinant human IL-17A is designated rhulL-17. A person skilled in the art knows how to express such proteins using starting materials and standard protocols as known in the art. However, in order to aid the W0 2014/122613 skilled artisan, unless otherwise stated, the following amino acid sequences may be used: cynomolgus monkey (cyno) IL-17A, SEQ ID NO: 79; cynolL-17F, SEQ ID NO: 80; rhesus monkey s) IL-17A, SEQ ID NO: 81; marmoset monkey (marmoset) IL- 17A, SEQ ID NO: 82; mouse (m) lL-17A, SEQ ID NO: 83; mlL-17F, SEQ ID NO: 84, rat lL-17A, SEQ ID NO: 85; human lL-17 receptor A (hulL-17RA), SEQ ID NO: 86. As is known to a person skilled in the art, the abovementioned sequences may vary slightly, i.e. due to originating from different population groups. In the examples, tool antibodies are also used e.g. for screening purposes. Such antibodies are standard dies and can be readily obtained by a person d in the art.

’IO The term "epitope" means a protein determinant capable of specific binding to an antibody. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics. Conformational and nonconformational epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents.

The term pe" refers to the dy class (e.g., IgM, IgE, IgG such as IgG1 or IgG4) that is provided by the heavy chain constant region genes. Isotype also includes modified versions of one of these classes, where modifications have been made to alter the Fc function, for example, to enhance or reduce effector functions or binding to Fc ors.

The term "human antibody", as used herein, is intended to include antibodies having variable regions in which both the framework and CDR regions are derived from sequences of human . Furthermore, if the antibody contains a constant region, the constant region also is derived from such human sequences, e.g., human germline sequences, or mutant versions of human germline ces or antibody containing sus framework ces derived from human framework sequences analysis, for e, as described in Knappik, et al. 2000, J Mol Biol 296:57-86).

A "humanized" antibody is an antibody that retains the reactivity of a non-human antibody while being less immunogenic in . This can be achieved, for instance, by retaining the non-human CDR regions and replacing the remaining parts of the dy with their human counterparts (i.e., the constant region as well as the W0 2014/122613 framework ns of the variable region). See, e.g., Morrison et al. 1984, Proc. Natl.

Acad. Sci. USA, 81:6851-6855; Morrison and Oi, 1988, Adv. lmmunol., 44:65-92; Verhoeyen et al. 1988, e, 239:1534-1536; Padlan 1991, Molec. lmmun., 28:489- 498; and Padlan 1994, Molec. lmmun., 31:169-217. Other examples of human engineering technology e, but are not limited to Xoma technology disclosed in US ,766,886.

The human antibodies of the disclosure may include amino acid residues not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by c mutation in vivo). However, the term "human 1O antibody", as used herein, is not intended to include antibodies in which CDR ces derived from the germline of another mammalian species, such as a mouse, have been d onto human framework sequences.

The term "human onal antibody" refers to antibodies displaying a single g specificity which have variable regions in which both the framework and CDR regions are derived from human sequences.

The term "recombinant human antibody", as used herein, includes all human antibodies that are prepared, expressed, created or isolated by inant means, such as antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human globulin genes or a hybridoma prepared therefrom, antibodies isolated from a host cell transformed to express the human antibody, e.g., from a transfectoma, antibodies isolated from a recombinant, combinatorial human antibody library, and antibodies prepared, expressed, created or isolated by any other means that involve splicing of all or a portion of a human immunoglobulin gene sequence to other DNA sequences. Such recombinant human dies have variable regions in which the framework and CDR regions are derived from human ne immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies can be subjected to in vitro mutagenesis (or, when an animal transgenic for human lg sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human dy germline repertoire in vivo.

W0 2014/122613 As used herein, "isotype" refers to the dy class (e.g., lgM, lgE, lgG, such as lgG1 or lgG4) that is provided by the heavy chain nt region genes.

The phrases "an antibody recognizing an antigen" and "an antibody specific for an antigen" are used interchangeably herein with the term "an antibody which binds specifically to an antigen".

As used herein, an antibody or a protein that "specifically binds to lL-17A polypeptide" is intended to refer to an antibody or protein that binds to human lL-17A polypeptide with a KB of 100nM or less, 10nM or less, 1nM or less, 100pM or less, or 10pM or less. An antibody that "cross-reacts with an antigen other than lL-17A " is intended to refer to an 1O antibody that binds that n with a KD of 10nM or less, 1 nM or less, or 100 pM or less. An antibody that "does not cross-react with a particular antigen" is intended to refer to an antibody that binds to that antigen, with a KB of 100 nM or greater, or a KB of 1 uM or grater, or a KB of 10 uM or greater. In certain embodiments, such antibodies that do not cross-react with the antigen exhibit essentially undetectable binding t these proteins in standard binding assays.

The term "Kassoc" or "Ka", as used herein, is ed to refer to the association rate of a particular antibody-antigen ction, whereas the term "Kdis" or ","Kd as used , is intended to refer to the dissociation rate of a particular antibody-antigen interaction.

The term "KD", as used herein, is intended to refer to the iation constant, which is obtained from the ratio of Kd to Ka (i.e. Kd/Ka) and is sed as a molar concentration (M). KD values for antibodies can be determined using methods well established in the art. A method for determining the KB of an antibody is by using surface plasmon resonance, or using a biosensor system such as a BiacoreTM system, well known to a person skilled in the art and operated e.g. as described in the Examples.

The tion of the binding of lL-17 to its receptor may be conveniently tested in various assays including such assays are described hereinafter in the text. By the term "to the same extent" is meant that the reference and the equivalent molecules exhibit, on a statistical basis, essentially identical lL-17 inhibitory activity in one of the assays referred to herein (see Examples). For example, lL-17 binding molecules of the disclosure typically have a half l inhibitory concentration (IC50), for the inhibition W0 2014/122613 of human lL-17 on lL-6 production induced by human lL-17 in human dermal fibroblasts, which is within +/-1O5, i.e. below 10 nM, more preferably 9, 8, 7, 6, 5, 4, 3 or 2 nM of that of, ably substantially the same as, the |C5o of the tive reference molecule when d e.g. as described in the Examples. Alternatively, the assay used may be an assay of competitive inhibition of binding of lL-17 by soluble lL-17 receptors and the lL-17 binding molecules of the disclosure.

As used herein, the term "Affinity" refers to the strength of ction between antibody and antigen at single nic sites. Within each antigenic site, the variable region of the antibody "arm" interacts through weak non-covalent forces with the antigen at 1O numerous sites; the more interactions, the stronger the affinity. As used herein, the term "high affinity" for an lgG antibody or fragment thereof (e.g., a Fab fragment) refers to an dy having a KD of 10'8 M or less, 10'9 M or less, or 10'10 M, or 10'11 M or less, or '12 M or less, or 10'13 M or less for a target antigen. However, high affinity binding can vary for other antibody isotypes. For example, high affinity binding for an lgM isotype refers to an antibody having a KB of 10'7 M or less, or 10'8 M or less.

As used herein, the term "Avidity" refers to an informative measure of the l ity or strength of the antibody-antigen complex. It is controlled by three major factors: antibody epitope affinity; the valence of both the antigen and antibody; and the structural arrangement of the interacting parts. Ultimately these factors define the specificity of the antibody, that is, the hood that the particular antibody is binding to a e antigen epitope.

As used herein, an antibody or protein that inhibits lL-17A binding to lL-17R is intended to refer to an antibody or protein that inhibits lL-17A binding to lL-17R with an |C5o of 10nM or less, preferably with an |C5o of 1nM or less, more preferably with an |C5o of 100pM, or less, as measured in an in vitro itive binding assay. Such assay is described in more details in the examples below.

As used herein, the term "IL-17A antagonist" or "IL-17A ng molecule" is intended to refer to an antibody or protein that inhibits lL-17A induced signaling activity through the lL-17R and thereby reduces or neutralizes lL-17A activity. This can be shown in a human cell assay such as the lL-17A dependent lL-6 or GRO-alpha production assay in human cells. Such assay is described in more detail in the Examples below. In some W0 2014/122613 ments, the antibodies or proteins of the disclosure t lL-17A dependent lL-6 or GRO-alpha production as measured in an in vitro human cell assay at an |C5o of 10nM or less, 1nM or less, or 100pM or less. Such an assay is described in more details in the Examples below. In some embodiments the antibodies or proteins of the disclosure inhibit antigen induced arthritis in in vivo assays in mice and rats. Such assays are bed in the Examples in more detail below.

As used , the term "ADCC" or "antibody dependent cell cytotoxicity" activity refers to cell depleting activity. ADCC activity can be measured by standard ADCC assay, well known to a person skilled in the art. 1O As used herein, the term "selectivity" for an antibody or protein of the disclosure refers to an antibody or protein that binds to a certain target ptide, but not to closely related ptides. The phrases "an dy recognizing an antigen" and ll antibody specific for an antigen" are used interchangeably herein with the term "an antibody which binds specifically to an n".

The term "nucleic acid" or "polynucleotide" refers to deoxyribonucleic acids (DNA) or ribonucleic acids (RNA) and polymers thereof in either single- or double-stranded form.

Unless specifically limited, the term encompasses nucleic acids containing known ues of l nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides.

(See, US. Pat. No. 8,278,036 to Kariko et a/., which discloses mRNA molecules with uridine replaced by pseudouridine, methods of synthesizing the same, and methods for the delivery of eutic proteins in vivo.) Methods for packaging mRNA can be used, for example, those disclosed in US. Pat. No. 8,278,036 to Kariko et al; and patent application W02013/O90186A1, to Moderna. Unless ise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively ed variants thereof (e.g., degenerate codon substitutions), s, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et a/., Nucleic Acid Res. 19:5081 (1991); Ohtsuka eta/., J. Biol. Chem. 260:2605-2608 (1985); and Rossolini et a/., Mol. Cell. Probes 8:91- 98 (1994)).

W0 2014/122613 As used herein, the term "subject" includes any human or nonhuman animal. The term "nonhuman animal" includes all vertebrates, e.g., mammals and non-mammals, such as an primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. As used herein, the terms "cyno" or "cynomolgus" refer to the cynomolgus monkey (Macaca fascicu/aris). As used , the terms "rhesus" or "rhesus macaque" refer to the rhesus macaque monkey a mulatta). As used herein, the term "marmoset" refers to a marmoset monkey.

As used herein, the term "treating" or "treatment" of any disease or disorder (i.e., rheumatoid arthritis) refers in one embodiment, to rating the disease or disorder 1O (i.e., slowing or arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In another embodiment "treating" or "treatment" refers to alleviating or ameliorating at least one physical parameter including those which may not be discernible by the patient. In yet another embodiment, "treating" or ment" refers to modulating the disease or disorder, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both. Methods for assessing treatment and/or prevention of disease are generally known in the art, unless specifically bed herein.

As used herein, "selecting" and "selected" in reference to a patient is used to mean that a particular patient is specifically chosen from a larger group of patients due to the particular patient having a predetermined criterion. Similarly, "selectively treating a patient" refers to providing treatment to a patient that is specifically chosen from a larger group of patients due to the particular t having a ermined criteria. Similarly, "selectively administering" refers to stering a drug to a t that is ically chosen from a larger group of patients due to the ular patient having a predetermined criterion.

As used herein, the term, "optimized" means that a nucleotide sequence has been altered to encode an amino acid sequence using codons that are preferred in the production cell or organism, generally a eukaryotic cell, for e, a cell of Pichia, a cell of Trichoderma, a Chinese Hamster Ovary cell (CHO) or a human cell. The optimized nucleotide sequence is ered to retain completely or as much as possible the amino acid sequence originally encoded by the ng nucleotide sequence, which is also known as the "parental" sequence. The optimized sequences W0 2014/122613 herein have been engineered to have codons that are preferred in CHO mammalian cells, however optimized expression of these sequences in other eukaryotic cells is also envisioned . The amino acid ces encoded by optimized nucleotide sequences are also referred to as optimized.

As used herein, the percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity = # of cal positions/total # of positions x 100), taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal ent of the two sequences. The comparison of sequences and determination of percent identity 1O between two sequences can be accomplished using a mathematical algorithm, as described below.

The percent identity between two amino acid sequences can be ined using the algorithm of E. Meyers and W. Miller 1988,Comput. Appl. ., 4:11-17, which has been incorporated into the ALIGN program (version 2.0), using a PAM12O weight residue table, a gap length penalty of 12 and a gap penalty of 4. Alternatively, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch 1970, J. Mol, Biol. 48:444-453 algorithm which has been orated into the GAP program in the GCG software package (available at http://www.gcg.com), using either a Blossom 62 matrix or a PAM25O matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.

The percent identity between two nucleotide amino acid sequences may also be determined using for example algorithms such as the BLASTN program for nucleic acid sequences using as defaults a word length (W) of 11, an expectation (E) of 10, M=5, N=4, and a comparison of both strands.

The terms "cross-block", -blocked", "cross-blocking" are used interchangeably herein to mean the ability of an antibody or other binding agent to interfere with the binding of other antibodies or binding agents to lL-17A in a standard itive binding assay.

The ability or extent to which an antibody or other binding agent, such as a protein comprising an antigen-binding n of an antibody, is able to interfere with the binding W0 2014/122613 of another dy or binding molecule to IL-17A, and therefore whether it can be said to cross-block ing to the disclosure, can be determined using standard competition binding assays. One suitable assay involves the use of the BiacoreTM technology (e.g. by using the BiacoreTM 3000 instrument (BiacoreTM, Uppsala, Sweden)), which can measure the extent of interactions using surface plasmon resonance logy. Another assay for measuring cross-blocking uses an ELISA- based approach. Further details on these methods are given in the Examples.

For example, the dies exemplified herein (i.e. XAB1, XABZ, XABS, XAB4 and XAB5) and proteins comprising an antigen-binding portion thereof will all -block" 1O one r. All of these antibodies target the same epitope on IL-17A. Other cross- ng antibodies would be anticipated to bind to the same, or a related, epitope.

According to the disclosure, a cross-blocking antibody or other binding agent, such as a protein comprising an antigen-binding portion of an antibody, according to the disclosure binds to IL-17A in the described BiacoreTM cross-blocking assay such that the recorded binding of the combination (mixture) of the antibodies or binding agents is between 80% and 0.1% (e.g. 80% to 4%) of the maximum tical binding, specifically between 75% and 0.1% (e.g. 75% to 4%) of the maximum theoretical g, and more specifically between 70% and 0.1% (e.g. 70% to 4%), and more specifically n 65% and 0.1% (e.g. 65% to 4%) of maximum theoretical binding (as defined above) of the two antibodies or binding agents in combination An antibody is defined as cross-blocking in the ELISA assay as described in the Examples, if the solution phase anti-IL-17A antibody is able to cause a reduction of between 60% and 100%, ically between 70% and 100%, and more specifically between 80% and 100%, of the IL-17A detection signal (i.e. the amount IL-17A bound by the coated antibody) as compared to the IL-17A detection signal obtained in the absence of the solution phase anti-IL-17A antibody (i.e. the positive control .

DETAILED DESCRIPTION OF THE INVENTION The present disclosure is based, in part, on the discovery of antibody molecules that specifically bind to homodimeric IL-17A and dimeric IL-17AF, but do not specifically bind to homodimeric IL-17F. The disclosure relates to both full IgG format W0 2014/122613 antibodies as well as proteins comprising an antigen-binding portion thereof, which will be further bed below.

Accordingly, the present disclosure provides antibodies as well as proteins comprising an antigen-binding portion thereof with binding lities that are surprisingly similar for several species, such as selected from one or more of cynomolgus, rhesus macaque, marmoset, rat, mouse or human, as well as pharmaceutical compositions, production methods, and methods of use of such antibodies and compositions.

Recombinant antibodies Antibodies of the disclosure include the human inant antibody XAB1 and antibody derivates XABZ, XABS, XAB4 and XAB5, which were derived, isolated and structurally terized by their full length heavy and light chain amino acid sequences as described in Table 1 below.

Table 1. Full length heavy and light chain amino acid sequences of XAB1, XABZ, XAB3, XAB4 and XAB5.

Antibody Full Length Heavy Chain Full Length Light Chain Amino acid ce Amino acid sequence XAB1 SEQ ID NO: 14 SEQ ID NO: 15 XABZ SEQ ID NO: 14 SEQ ID NO: 26 XA83 SEQ ID NO: 14 SEQ ID NO: 36 XAB4 SEQ ID NO: 14 SEQ ID NO: 44 XAB5 SEQ ID NO: 14 SEQ ID NO: 54 SEQUENCE 100% 97% IDENTITY The corresponding variable regions, VH and VL amino acid sequences of such isolated dies XAB1, XABZ, XABS, XAB4 and XAB5 of the disclosure are shown in Table 2 below.

W0 2014/122613 Table 2. Variable heavy and light chain amino acid sequences of XAB1, XABZ, XAB3, XAB4 and XAB5.

-——Antibody Variable Heavy Chain Variable Light Chain XA83 SEQ ID NO: 12 SEQ ID NO: 35 XAB4 SEQ ID NO: 12 SEQ ID NO: 43 XAB5 SEQ ID NO: 12 SEQ ID NO: 53 SEQUENCE 100% 94% IDENTITY Other antibodies of the disclosure include those having amino acids that have been d by amino acid deletion, insertion or substitution, yet have at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 98%, 99% or 100% identity to the antibodies described above, in particular in the CDR regions depicted in the sequences described above. In some ments, the antibody of the disclosure is a mutant variant of any one of XAB1, XAB2, XAB3, XAB4 and XAB5, wherein said mutant variant antibody include mutant amino acid sequences wherein no more than 1, 2, 3, 4 or 5 amino acids have been mutated by amino acid deletion, insertion or substitution in the CDR s when compared with the CDR regions depicted in the sequences bed above.

Full length light and heavy chains nucleotide coding sequences of XAB1, XAB2, XAB3, XAB4 and XAB5 are shown in Table 3 below.

Table 3. Full length heavy and light chain DNA coding sequences.

-——Antibody Full Length Heavy Chain Full Length Light Chain XAB1 SEQ ID NO: 18, SEQ ID NO: 19 SEQ ID NO: 31, SEQ ID NO: 51 XABZ SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 31, SEQ ID NO: 32 W0 2014/122613 SEQ ID NO: 18, SEQ ID NO: 38, SEQ ID NO: 31, SEQ ID NO: 40 SEQ ID NO: 51 SEQ ID NO: 18, SEQ ID NO: 46, SEQ ID NO: 31, SEQ ID NO: 48, SEQ ID NO: 51 SEQ ID NO: 52 SEQ ID NO: 18, SEQ ID NO: 56, SEQ ID NO: 31, SEQ ID NO: 58 SEQ ID NO: 51 Variable light and heavy chains nucleotide coding ces of XAB1, XAB2, XABS, XAB4 and XAB5 are shown in Table 4 below.

Table 4. Variable heavy and light chain amino acid sequences DNA coding sequences.

-——Antibody Variable Heavy Chain Variable Light Chain XAB1 SEQ ID NO: 16, SEQ ID NO: 17 SEQ ID NO: 29, SEQ ID NO: 49 XAB2 SEQ ID NO: 16, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 30 SEQ ID NO: 49 XAB3 SEQ ID NO: 16, SEQ ID NO: 37, SEQ ID NO: 29, SEQ ID NO: 39 SEQ ID NO: 49 XAB4 SEQ ID NO: 16, SEQ ID NO: 45, SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 49 SEQ ID NO: 50 XAB5 SEQ ID NO: 16, SEQ ID NO: 55, W0 2014/122613 2014/058854 SEQ ID NO: 29, SEQ ID NO: 57 SEQ ID NO: 49 Other nucleic acids encoding antibodies of the disclosure include nucleic acids that have been mutated by nucleotide deletion, insertion or substitution, yet have at least 60, 70, 80, 90, 95 or 100 percent identity to the CDR corresponding coding regions depicted in the sequences described above or in Table 5 and Table 6 below.

In some embodiments, it include variant nucleic acids wherein no more than 1, 2, 3, 4 or nucleotides have been changed by nucleotide deletion, insertion or substitution in the CDR coding s with the CDR coding regions ed in the sequences bed above or in Table 5 and Table 6 below.

For antibodies that bind to the same epitope, the VH, VL, full length light chain, and full 1O length heavy chain sequences (nucleotide sequences and amino acid sequences) can be "mixed and matched" to create other anti-lL-17A binding les of the disclosure. lL-17A binding of such "mixed and d" antibodies can be tested using the binding assays described above or other conventional binding assays (e.g., ELISAs). When these chains are mixed and d, a VH sequence from a particular VHNL pairing should be replaced with a urally similar VH ce. Likewise a full length heavy chain sequence from a particular full length heavy chain /full length light chain pairing should be replaced with a structurally similar full length heavy chain ce.

Likewise, a VL sequence from a particular VH/VL pairing should be replaced with a structurally similar VL sequence. Likewise a full length light chain sequence from a 2O particular full length heavy chain / full length light chain pairing should be replaced with a structurally similar full length light chain ce. Accordingly, in one aspect, the disclosure provides an isolated recombinant antibody or protein comprising an antigen- binding portion thereof having: a heavy chain variable region comprising an amino acid sequence ed from the group consisting of SEQ ID NO: 12 and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 13, 25, 35, 43 and 53; wherein said heavy and light chain regions are selected such that the antibody specifically binds to lL-17A.

Examples of the amino acid sequences of the VH CDR1s (also called HCDR1 or HCDR1’ depending of the CDR definition that is used), VH CDR2s (also called HCDR2 W0 2014/122613 or HCDR2’ depending of the CDR definition that is used), VH CDR3s (also called HCDR1 or HCDR1’ depending of the CDR definition that is used), VL CDR1s (also called LCDR1 or LCDR1’ depending of the CDR definition that is used), VL CDR2s (also called LCDR2 or LCDR2’ depending of the CDR tion that is used), VL CDR3s (also called HCDR3 or HCDR3’ depending of the CDR definition that is used) of some antibodies and proteins comprising an antigen-binding portion thereof according to the disclosure are shown in Table 5 and Table 6.

In Table 5, the CDR regions of some antibodies of the disclosure are delineated using the Kabat system (Kabat, E. A., et al. 1991, ces of ns of Immunological Interest, Fifth n, US. Department of Health and Human Services, NIH Publication No. 91-3242, see also Zhao&Lu 2009, Molecular Immunology 47:694-700) For the ease of reading, when CDR regions are delineated according to Kabat definition, they are called hereafter HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3 respectively.

Table 5. CDR regions of XAB1, XABZ, XAB3, XAB4 and XAB5 according to Kabat definition.

Original HCDR1 HCDR2 HCDR3 LCDR1 LCDR2 LCDR3 XA81 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID XABZ SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID XABS SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 7 NO: 8 NO: 3 NO: 34 NO: 23 NO: 11 XAB4 SEQID SEQID SEQID SEQID SEQID SEQID XAB5 SEQID SEQID SEQID SEQID SEQID SEQID CONSENSUS SEQID SEQID SEQID SEQID SEQID SEQID SEQUENCE 100% 100% 100% 64 % 86% 89% IDENTITY The consensus sequences SEQ ID NO: 73, SEQ ID NO: 74 and SEQ ID NO: 75 comprise a number of variable amino acids, designated X. Based on sequence alignment of the sequences for XAB2 to XAB5, the four variable amino acids in SEQ ID NO: 73 can advantageously be selected according to the following: The first variable amino acid (X1) can be ed from the group consisting of Gly (G) and Val (V); the second le amino acid (X2) can be selected from the group consisting of Tyr (Y), Asn (N) and lie (I); the third variable amino acid (X3) can be selected from the group consisting of Trp (W) and Ser (S); and the fourth variable amino acid (X4) can be selected from the group consisting of Glu (E) and Ala (A). SEQ ID NO: 9 has a sequence identity of 91% compared to SEQ ID NO: 22 and a sequence identity of 73% compared to SEQ ID NO: 34 and SEQ ID NO: 42. SEQ ID NO: 22 has a sequence identity of 64% compared to SEQ ID NO: 34 and SEQ ID NO: 42. SEQ ID NO: 34 has a sequence ty of 91% ed to SEQ ID NO: 42.

Similarly, the one variable amino acid in SEQ ID NO: 74 can advantageously be selected ing to the following: X1 can be selected from the group consisting of Asn (N) and Gln (Q). SEQ ID NO: 10 has a sequence identity of 86% compared to SEQ ID NO: 23.

The one variable amino acid in SEQ ID NO: 75 can advantageously be selected according to the following: X1 can be selected from the group consisting of Asn (N) and Asp (D). SEQ ID NO: 11 has a sequence identity of 89% compared to SEQ ID NO: 24.

In Table 6, the CDR regions of some antibodies of the sure are delineated using the Chothia system, Al-Lazikani et al. 1997, J. Mol. Biol. 273:927-948. For ease of g, when the CDR regions are ated according to Chothia definition, they are called hereafter HCDR1’, HCDR2’, HCDR3’, LCDR1’, LCDR2’, LCDR3’ respectively.

Table 6. CDR regions from XAB1, XABZ, XAB3, XAB4 and XAB5 according to Chothia definition.

W0 2014/122613 XAB1 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID XAB2 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID XAB3 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID XAB4 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID XAB5 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID CONSENSUS SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID ------SEQUENCE 100% 100% 100% 43% 100% 83% The consensus sequences SEQ ID NO: 71 and SEQ ID NO: 72 comprise a number of variable amino acids, designated X. Based on sequence alignment of the sequences for XAB2 to XAB5, the four variable amino acids in SEQ ID NO: 71 can advantageously be selected according to the ing: the first variable amino acid (X1) can be selected from the group consisting of Gly (G) and Val (V), the second variable amino acid (X2) can be selected from the group consisting of Tyr (Y), Asn (N) and lie (I); the third variable amino acid (X3) can be ed from the group consisting of Trp (W) and Ser (S); and the fourth variable amino acid (X4) can be selected from the group ting of Glu (E) and Ala (A). SEQ ID NO: 4 has a sequence identity of 86% compared to SEQ ID NO: 20 and a sequence identity of 57% compared to SEQ ID NO: 33 and SEQ ID NO: 41. SEQ ID NO: 20 has a sequence identity of 43% compared to SEQ ID NO: 33 and SEQ ID NO: 41. SEQ ID NO: 33 has a sequence ty of 86% compared to SEQ ID NO: 41.

Similarly, the one variable amino acid in SEQ ID NO: 72 can advantageously be selected according to the following: X1 can be selected from the group consisting of Asn (N) and Asp (D). SEQ ID NO: 6 has a sequence identity of 86% compared to SEQ ID NO: 21.

W0 22613 Given that each of these antibodies can bind to IL-17A and that antigen-binding icity is provided primarily by the CDR1, 2 and 3 regions, the VH CDR1, 2 and 3 sequences and VL CDR1, 2 and 3 sequences can be "mixed and d" (i.e., CDRs from different antibodies can be mixed and matched, each antibody containing a VH CDR1, 2 and 3 and a VL CDR1, 2 and 3 create other anti-IL-17A binding molecules of the disclosure). IL-17A binding of such "mixed and matched" antibodies can be tested using the binding assays described above and in the Examples or other conventional assays (e.g., ELISAs). When VH CDR sequences are mixed and matched, the CDR1, CDR2 and/or CDR3 sequence from a ular VH sequence should be ed with a ’IO structurally similar CDR sequence(s). Likewise, when VL CDR sequences are mixed and matched, the CDR1, CDR2 and/or CDR3 sequence from a particular VL sequence should be replaced with a structurally similar CDR sequence(s). It will be readily apparent to the ordinarily skilled n that novel VH and VL sequences can be d by substituting one or more VH and/or VL CDR region(s) sequence(s) with structurally similar sequences from the CDR sequences shown herein for onal antibodies of the present disclosure.

In one embodiment, an isolated recombinant antibody, or a protein comprising an n-binding portion thereof, has: a heavy chain variable region CDR1 according to SEQ ID NO: 7; a heavy chain variable region CDR2 according to SEQ ID NO: 8; a heavy chain variable region CDR3 according to SEQ ID NO: 3; a light chain variable region CDR1 sing an amino acid sequence selected from the group consisting of SEQ ID NO: 9, 22, 34, 42, and 73, and preferably selected from the group consisting of selected from the group consisting of SEQ ID NO: 9, 22, 34, 42; a light chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 10, 23, and 74, and preferably selected from the group ting of selected from the group consisting of SEQ ID NO: 10 and 23; and a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 11, 24, and 75 and preferably selected from the group consisting of selected from the group consisting of SEQ ID NO: 11 and 24; wherein said CDR regions are selected so that the antibody or protein of the disclosure specifically binds to IL-17A W0 2014/122613 In another embodiment, an isolated inant antibody, or a protein sing an antigen-binding portion f has: a heavy chain variable region HCDR1’ according to SEQ ID NO: 1; a heavy chain variable region HCDR2’ according to SEQ ID NO: 2; a heavy chain variable region HCDR3’ according to SEQ ID NO: 3; a light chain variable region LCDR1’ comprising an amino acid ce selected from the group consisting of SEQ ID NO: 4, 20, 33, 41, and 71, and preferably selected from the group consisting of selected from the group consisting of SEQ ID NO: 4, 20, 33, 41; a light chain variable region LCDR2’ according to SEQ ID NO: 5; and a light chain variable region LCDR3’ comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1O 6, 21, and 72, and preferably selected from the group consisting of selected from the group consisting of SEQ ID NO: 6 and 21; wherein said CDR s are selected so that the antibody or protein of the sure specifically binds to IL-17A.

In certain embodiments, the antibody or protein comprising an antigen-binding portion thereof comprises either SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3; SEQ ID NO: 12; or c) SEQ ID NO: 14.

As used herein, a human antibody comprises heavy or light chain variable regions or full length heavy or light chains that are "the t of" or "derived from" a particular germline sequence if the variable regions or full length chains of the antibody are obtained from a system that uses human germline immunoglobulin genes. Such s include immunizing a transgenic mouse carrying human immunoglobulin genes with the antigen of interest or screening a human globulin gene library displayed on phage with the antigen of st. A human dy that is "the product of" or "derived from" a human germline globulin sequence can be identified as such by comparing the amino acid sequence of the human antibody to the amino acid sequences of human germline immunoglobulins and selecting the human germline immunoglobulin sequence that is closest in sequence (i.e., greatest % identity) to the sequence of the human antibody. A human antibody that is "the product of" or "derived from" a particular human germline immunoglobulin sequence may contain amino acid differences as compared to the germline sequence, due to, for example, naturally occurring somatic mutations or intentional introduction of site-directed mutation.

However, a selected human antibody typically is at least 90% cal in amino acids sequence to an amino acid sequence encoded by a human germline immunoglobulin W0 22613 gene and contains amino acid residues that identify the human antibody as being human when compared to the germline immunoglobulin amino acid sequences of other species (e.g., murine ne sequences). In n cases, a human antibody may be at least 60%, 70%, 80%, 90%, or at least 95%, or even at least 96%, 97%, 98%, or 99% identical in amino acid sequence to the amino acid sequence encoded by the germline immunoglobulin gene. Typically, a human antibody derived from a particular human germline sequence will display no more than 10 amino acid differences from the amino acid sequence encoded by the human germline immunoglobulin gene. In n cases, the human antibody may display no more than 5, or even no more than 4, 3, 2, or 1 1O amino acid difference from the amino acid ce encoded by the germline immunoglobulin gene.

In the present disclosure, an epitope on lL-17A has been identified that is ularly preferred as a target for the binding of potentially therapeutic antibodies. This epitope is bound by XAB1, and the t antibodies XAB2, XAB3, XAB4 and XAB5 which have been ped by modification of the sequence of XAB1. This epitope is found on the lL-17A sequence, between residues Arg 78 and Trp 90.

The epitope may be considered to comprise the following most preferred amino acid residues from within the lL-17A: Arg 78, Glu 80, Trp 90. In addition, the ing amino acid residues are also preferred: Tyr 85, Arg 124. Other important amino acid residues are Pro 82, Ser 87, Val 88. Further contributing amino acid residues are Val 45*, Leu 49, Asp 81, Glu 83, Pro 86, Pro 130, Phe 133, Lys 137*, where amino acids marked with (*) designate residue contributed by the second lL-17A subunit of the homodimer lL-17A.

Antibodies that target this epitope on lL-17A have been shown to block the binding of lL-17A to its receptor to inhibit lL-17A mediated effects in vitro, and to reduce the severity of an mental in vivo model of antigen induced arthritis. In addition, antibodies that bind to this epitope have unexpectedly been shown to inhibit the in vitro effects mediated by the lL-17AF heterodimer, and also to retain an ctedly high affinity for lL-17A and lL-17AF derived from mouse and other species variations of the target molecule.

W0 2014/122613 Thus, this epitope is especially preferred since it is also unexpectedly ved in an accessible format within the structure of the lL-17AF heterodimer. Accordingly, preferred antibodies of the disclosure will also bind to lL-17AF heterodimers. Without wishing to be bound by theory, it is anticipated that the structure of the lL-17AF heterodimer is sufficiently r, to lL-17A, or that ction with the antibodies of the disclosure renders it sufficiently similar to lL-17A, that binding may still occur.

This is unexpected because structural analyses based on the available structures for lL- 17A, lL-17F and interactions between these molecules and antibodies or receptors that have been obtained by X-ray crystallography shed in the art and conducted by the 1O inventors) in combination with in silico predictions, suggested that binding to, or crossreactivity of the antibodies of the disclosure with lL-17AF would not necessarily occur.

More specifically, the N-terminal region of the lL-17F monomeric subunit of the heterodimer was predicted to sterically hamper the binding of antibodies of the disclosure to the F heterodimer. The expectation was thus that there would not be significant cross-reactivity for the antibodies with lL-17AF.

However, despite these tions, we have determined that cross-binding to lL-17AF by the disclosed antibodies does occur. This may in fact be advantageous for a number of reasons. As discussed above, lL-17AF is also implicated as a flammatory cytokine and may be involved in many of the same pathological conditions or undesirable biological events as described or suspected for lL-17A. The antibodies of the disclosure may therefore be especially valuable therapeutically because they can target or ere with both lL-17A and lL-17AF. er, the present inventors have demonstrated that this binding between the dies of the sure and lL-17AF also correlates to an inhibition of the biological activity of lL-17AF as observed in in vitro assays. Accordingly, the antibodies of the disclosure not only efficiently target and antagonize/neutralize the activity of , but also lL-17AF activity as well.

A further unexpected consequence of the work conducted by the t ors is as follows. The affinity maturation of the original ‘parent’ antibody XA81 has also resulted in a set of antibodies retaining a high affinity, or an improved affinity for lL-17A variants W0 2014/122613 derived from other species such as cynomolgus, rhesus macaque, marmoset, rat, or mouse.

This is unexpected e in striving to improve the affinity of the antibodies of the disclosure for human lL-17A it would not be expected to also improve the affinity of the ing antibodies for species variants of lL-17A. In fact, the opposite might normally be expected. s to e antibody affinity for a specific species variant (i.e. human) of a target antigen would y be expected to reduce affinity for other species variants of that antigen. The concept of the species variant (or homolog/paralog) recognizes a common ancestry for a given species, but accepts that 1O divergence has taken place over the course of evolutionary history. Accordingly, even where there is a good degree of ce conservation between ts of a particular molecule that have been identified in different species, it cannot be assumed that an improved affinity for one species variant will have an improvement on the affinity for another species variant. In fact, the ence between the sequences for ent species generally leads to the ation that improvement in affinity for one variant will be more likely to lead to a reduction (or even abolishment) of the binding affinity for another species variant. The sequence identity between mouse and human lL-17A is only 62% (Moseley et al. 2003, Cytokine & Growth Factor Reviews 14:155—174).

However, in the present case this was not observed and the antibody variants ted by the ors retained high affinity for lL-17A variants from other species.

This is useful because during the work necessary to develop a candidate antibody molecule as a useful eutic molecule a variety of tests and assays may be required to be carried out in other species or on cells, molecules or systems comprising components of, or derived from other species (such as cynomolgus, rhesus macaque, marmoset, rat, or mouse). This makes the antibodies of the disclosure are especially suitable for further development.

Accordingly, antibodies and proteins comprising an antigen-binding portion thereof as disclosed herein may share a range of desirable properties including, high affinity for lL- 17A, cross-reactivity with lL-17A from other species such as mouse, rat, cynomolgus and marmoset, lack of cross-reactivity for other lL-17 isotypes such as lL-17F, lack of cross-reactivity for other cytokines (such as human or mouse cytokines), cross-reactivity with heterodimeric lL-17AF, the ability to block g of lL-17A to its receptor such as W0 2014/122613 lL-17RA, the ability to inhibit or neutralize lL-17A induced biological effects such as the stimulation of lL-6 or GRO-alpha secretion, and/or the ability to inhibit in vivo effects mediated by lL-17A (and/or lL-17AF) such as the swelling that is observed in antigen induced arthritis models.

The antibodies and proteins comprising an antigen-binding portion thereof as disclosed herein have also been shown to provide a slow elimination of the antibody-IL17A complex, a slow turnover of the ligand and a long duration of |L17A capture. Further advantageous features of these antibodies and proteins are provided in the detailed embodiments. 1O Homologous antibodies In yet another embodiment, an antibody or n comprising an antigen-binding portion thereof as disclosed herein has full length heavy and light chain amino acid sequences; full length heavy and light chain nucleotide sequences, variable region heavy and light chain nucleotide sequences, or variable region heavy and light chain amino acid sequences, or all 6 CDR s amino acid sequences or nucleotide coding sequences that are homologous to the amino acid or nucleotide sequences of the antibodies XABl, XABZ, XABS, XAB4 and XAB5 described above, in particular in Table 1, and wherein the antibodies or proteins of the disclosure retain the desired onal properties of the original XABl, XABZ, XABS, XAB4 and XAB5 antibodies. d functional properties of the original XABl, XABZ, XABS, XAB4 and XAB5 antibodies may be selected from the group ting of: (i) binding ty to lL-17A (specific binding to lL-17A), for example, a KD being 1nM or less, 100pM or less, or 10pM or less, as measured in the BiacoreTM assay, e.g. as described the Examples; (ii) competitive inhibition of lL-17R binding to lL-17A, for example, an |C5o being 10nM or less, or 1nM or less, or 100pM or less, as ed in an in vitro competitive g assay, e.g. as described in the es; (iii) inhibition of lL-17A dependent activity, for example production of lL-6 or GRO-alpha, for example, an |C5o being 10nM or less, or 1nM or less, or W0 2014/122613 100pM or less, as ed in a cellular assay as described in the Examples; (iv) inhibition of the effects ed, for example knee swelling, as ed in an in vivo antigen induced arthritis assay as described in the Examples; (v) cross-reactivity with cynomolgus, rhesus macaque, rat, or mouse lL-17A polypeptide; (vi) cross-reactivity with human or mouse lL-17AF polypeptide; (vii) binding affinity to lL-17AF (specific binding to lL-17AF), for example, a KD being 1nM or less, 100pM or less, or 10pM or less, as measured in the 1O BiacoreTM assay, e.g. as described in the Examples; (viii) inhibition of lL-17AF, for example, an |C5o being 200nM or less, 150 nM or less, or 100 nM or less, as measured in an in vitro competitive binding assay as described in the Examples; (ix) suitable properties for drug development, in particular, it is stable and does not aggregate at in a ation at high concentration, i.e., above 50mg/ml.

For example, the disclosure relates to gous antibodies of XABl, XABZ, XABS, XAB4 and XAB5 (or a n comprising an antigen-binding portion thereof), comprising a le heavy chain (VH) and a variable light chain (VL) sequences where the CDR sequences, i.e. the 6 CDR regions; HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3 or HCDR1’, HCDR2’, HCDR3’, LCDR1’, LCDR2’, LCDR3’, share at least 60, 70, 90, 95 or 100 percent sequence identity to the corresponding CDR sequences of at least one antibody of XABl, XABZ, XABS, XAB4 and XABS, wherein said gous antibody or antigen-binding fragment thereof, such as a protein sing an antigen-binding portion thereof, specifically binds to lL-17A, and the antibody or protein exhibits at least one of the following functional ties: it inhibits g of lL-17A to its receptors, it inhibits lL-17A dependent lL-6 or GRO-alpha production in cellular assays, or inhibition of the effects observed in an in vivo antigen d arthritis assay. In a related specific embodiment, the homologous antibody or protein binds to lL-17A with a KB of 1nM or less and inhibits binding of lL-17A to its receptors as measured in an in vitro competitive binding assay with an |C5o of 1nM or less. The CDRs of XABl, XABZ, XABS, XAB4 and XAB5 are defined in the above Table and Table 6.

W0 2014/122613 The disclosure further relates to homologous antibodies of XABl, XABZ, XABS, XAB4 and XAB5 (or n-binding fragments thereof, such as a protein comprising an antigen-binding portion f) sing a heavy chain variable region and a light chain variable region that are at least 80%, 90%, or at least 95% or 100% identical to the corresponding heavy and light chain variable regions of any one of the antibodies XABl, XABZ, XABS, XAB4 or XABS; the homologous antibody or protein specifically binds to lL-17A, and it exhibits at least one of the following functional properties: it inhibits g of lL-17A to its receptor(s), it inhibits lL-17A dependent lL-6 or GRO- alpha production in cellular assays, or inhibition of the effects observed in an in vivo 1O antigen induced arthritis assay. In a related specific embodiment, the homologous antibody or n binding fragment f, such as a protein comprising an antigenbinding portion f binds to lL-17A with a KB of 1nM or less and inhibits binding of lL-17A to its receptor(s), as ed in an in vitro itive binding assay with an |C5o of 1nM or less. The VH and VL amino acid sequences of XABl, XABZ, XABS, XAB4 and XAB5 are defined in the above Table 2.

In another example, the disclosure s to homologous dies of XABl, XABZ, XABS, XAB4 and XAB5 (or antigen-binding fragments thereof, such as a protein sing an antigen-binding portion thereof) comprising a full length heavy chain and a full length light chain, wherein: the variable heavy chain is encoded by a nucleotide sequence that is at least 80%, at least 90%, at least 95%, or 100% identical to the corresponding coding nucleotide sequence of the variable heavy and light chains of XABl, XABZ, XABS, XAB4 and XABS, the homologous antibody or antigen-binding fragments thereof, such as a protein comprising an antigen-binding portion thereof, specifically binds to lL-17A, and it exhibits at least one of the following functional properties: it inhibits binding of lL-17A to its receptor(s), it inhibits lL-17A dependent production of lL-6 or GRO-alpha in cellular assays, or inhibition of the effects observed in an in vivo antigen induced arthritis assay. In a related specific embodiment, the homologous antibody or antigen-binding fragments thereof, such as a protein sing an antigen-binding portion thereof binds to lL-17A with a KB of 1nM or less and inhibits binding to lL-17A as measured in an in vitro competitive binding assay with an |C5o of 1nM or less. The coding nucleotide sequences of the variable regions of XABl, XABZ, XABS, XAB4 and XAB5 can be derived from the Table 3 showing the full length coding nucleotide sequences of XABl, XABZ, XABS, XAB4 and XAB5 and Table W0 2014/122613 2014/058854 2 showing the amino acid ces of the variable regions of XABi, XAB2, XABS, XAB4 and XAB5.

In various embodiments, the antibody or n-binding fragment thereof, such as a protein comprising an antigen-binding portion of an antibody, may exhibit one or more, two or more, three or more, or four or more of the desired functional properties sed above. The antibody or protein of the sure can be, for example, a human antibody, a humanized antibody or a chimeric antibody. In one embodiment, the antibody or protein is a fully human silent antibody, such as a fully human silent lgG1 antibody. 1O ed effector functions can be obtained by mutation in the Fc constant part of the antibodies and have been described in the Art: Strohl 2009 (LALA & N297A); Baudino 2008, D265A (Baudino et al. 2008, J. Immunol. 181:6664-69, Strohl, CO 2009, Biotechnology 20:685-91). Examples of silent lgG1 dies comprise the so-called LALA mutant comprising L234A and L235A mutation in the lgG1 Fc amino acid ce. r example of a silent lgG1 antibody comprises the D265A mutation.

The D265A mutation can also preferably be ed with the P329A mutation (DAPA). r silent lgG1 antibody comprises the N297A mutation, which results in aglycosylated or non-glycosylated antibodies.

Antibodies with mutant amino acid sequences can be obtained by mutagenesis (e.g., site-directed or PCR-mediated mutagenesis) of the coding nucleic acid molecules, followed by testing of the encoded altered antibody for retained function (i. e., the functions set forth above) using the functional assays described herein.

Antibodies with conservative modifications In certain embodiments, an antibody (or a protein comprising antigen-binding portion thereof) of the disclosure has a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences (or HCDR1’, HCDR2’ and HCDR3’) and a light chain variable region comprising LCDR1, LCDR2, and LCDR3 ces (or LCDR1’, LCDR2’, and LCDR3’), wherein one or more of these CDR sequences have specified amino acid sequences based on the antibodies XABi, XAB2, XABS, XAB4 or XAB5 described herein or conservative modifications f, and wherein the antibody or W0 2014/122613 protein retains the desired functional properties of the anti-lL-17A antibodies of the disclosure.

As used herein, the term "conservative sequence cations" is intended to refer to amino acid substitutions in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These es e amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, , 1O leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, phan, ine). Thus, one or more amino acid residues within the CDR regions of an antibody of the disclosure can be ed with other amino acid residues from the same side chain family, and the altered dy can be tested for retained function using the functional assays described herein.

Modifications can be introduced into an dy as disclosed herein by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis.

Engineered and modified antibodies Other antibodies and antigen-binding fragments, such as proteins comprising an antigen-binding portion thereof can be prepared using an antibody having one or more of the VH and/or VL sequences of XABl, XABZ, XABS, XAB4 or XAB5 shown above as starting material to engineer a modified antibody, which ed antibody may have altered properties from the starting antibody. An antibody can be engineered by ing one or more residues within one or both variable regions (i. e., VH and/or VL), for example within one or more CDR regions and/or within one or more framework regions. Additionally or alternatively, an antibody can be engineered by modifying residues within the constant region(s), for e to alter the effector function(s) of the antibody.

W0 2014/122613 One type of variable region engineering that can be performed is CDR grafting.

Antibodies ct with target antigens predominantly h amino acid residues that are located in the six heavy and light chains complementarity determining s (CDRs). For this reason, the amino acid sequences within CDRs are more diverse between individual antibodies than sequences e of CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, it is possible to express inant antibodies that mimic the properties of specific naturally occurring antibodies by constructing expression vectors that include CDR sequences from the specific naturally occurring antibody grafted onto framework sequences from a different 1O antibody with different ties (see, e.g., Riechmann, L. et al. 1998, Nature 332323- 327; Jones, P. et al. 1986, Nature 321:522-525; Queen, C. et al. 1989, Proc. Natl. Acad.

See. U.S.A. 29-10033; US. Patent No. 5,225,539 to Winter, and US. Patent Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.) Accordingly, another embodiment of the disclosure pertains to an isolated recombinant afted anti-lL-17A antibody, comprising the 6 CDR regions of any one of XAB1, XAB2, XAB3, XAB4 or XAB5 as defined in Table 5 or Table 6, yet containing different framework sequences from the original antibodies.

Such ork ces can be obtained from public DNA databases or published references that include germline dy gene sequences. For example, germline DNA sequences for human heavy and light chains variable region genes can be found in the "VBase" human germline sequence se (available on the Internet at www.mrc- cpe.cam.ac.uk/vbase), as well as in Kabat, E. A., et al. 1991, Sequences of Proteins of Immunological st, Fifth Edition, US. Department of Health and Human Services, NIH Publication No. 91-3242; Tomlinson, l. M, et al. 1992, J. Mol. Biol. 227:776-798; and Cox, J. P. L. et al. 1994, Eur. J lmmunol. 24:827-836.

Examples of framework sequences are those that are structurally similar to the framework sequences used in any one of XAB1, XAB2, XAB3, XAB4 or XAB5. The VH CDR1, 2 and 3 sequences, and the VL CDR1, 2 and 3 sequences, can be grafted onto framework regions that have the identical sequence as that found in the germline immunoglobulin gene from which the framework sequence derive, or the CDR sequences can be grafted onto framework regions that contain one or more mutations as compared to the germline sequences. For e, it has been found that in certain W0 2014/122613 instances it is beneficial to mutate residues within the framework regions to in or enhance the antigen binding ability of the antibody (see e.g., US. Patent Nos. ,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al).

Another type of variable region modification is to mutate amino acid residues within the VH and/or VL CDR1, CDR2 and/or CDR3 regions to thereby improve one or more binding properties (e.g., affinity) of the antibody of interest, known as "affinity maturation. Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to uce the mutation(s) and the effect on antibody binding, or other functional property of interest, can be evaluated in in vitro or in vivo assays as described 1O herein and provided in the Examples. Therefore, in one ment, the disclosure s to affinity matured antibodies derived from one of XAB1, XAB2, XAB3, XAB4 or XAB5 antibodies. Conservative modifications (as discussed above) can be introduced.

The mutations may be amino acid substitutions, additions or ons. Moreover, typically no more than one, two, three, four or five residues within a CDR region are altered. For example, an antibody of the disclosure is an affinity-matured antibody sing the 6 CDRs of one of XAB1, XAB2, XAB3, XAB4 or XAB5 and wherein no more than one, two, three, four or five residues within a CDR region are altered.

Accordingly, in another embodiment, the disclosure provides isolated engineered anti- lL-17A antibodies sing a heavy chain variable region and a light chain variable region which are identical to the ponding heavy and light chain variable regions of at least one of XAB1, XAB2, XAB3, XAB4 or XAB5 antibodies except that the heavy and/or light chain amino acid sequences of said ered antibodies contain one, two, three, four or five amino acid substitutions, deletions or additions as compared to the original sequences. ng antigen-binding domains into alternative frameworks or scaffolds A wide variety of antibody/immunoglobulin frameworks or scaffolds can be employed so long as the resulting ptide includes at least one binding region of XAB1, XAB2, XAB3, XAB4 or XAB5, which specifically binds to lL-17A. Such frameworks or lds include the 5 main idiotypes of human immunoglobulins, or fragments thereof (such as those disclosed elsewhere herein), and include immunoglobulins of other animal species, preferably having humanized aspects. Single heavy-chain antibodies such as W0 2014/122613 2014/058854 those identified in camelids are of particular interest in this regard. Novel frameworks, scaffolds and fragments continue to be discovered and developed by those skilled in the art.

In one aspect, the disclosure pertains to generating non-immunoglobulin based antibodies or ns sing an antigen-binding portion f using non- immunoglobulin scaffolds onto which CDRs of the disclosure can be grafted. Known or future non-immunoglobulin frameworks and scaffolds may be employed, as long as they comprise a binding region specific for the target protein of SEQ ID NO: 76. Such compounds are referred herein as "polypeptides comprising a target-specific g 1O region". Examples of non-immunoglobulin ork are r described in the sections below (camelid antibodies and non-antibody scaffold).

Came/id antibodies Antibody proteins obtained from members of the camel and dromedary (Came/us bactrianus and Cale/us dromaderius) family including new world members such as llama species (Lama paccos, Lama glama and Lama vicugna) have been characterized with respect to size, structural xity and antigenicity for human subjects. Certain lgG antibodies from this family of mammals as found in nature lack light chains, and are thus structurally distinct from the typical four chain quaternary structure having two heavy and two light chains, for antibodies from other animals. See PCT Publication No.

WO 94/04678.

A region of the camelid antibody which is the small single variable domain identified as VHH can be obtained by genetic engineering to yield a small protein having high affinity for a target, resulting in a low molecular weight antibody-derived protein known as a id nanobody". See US. Patent No. 5,759,808 issued June 2, 1998; see also Stijlemans, B. et al. 2004, J Biol Chem 279: 261; Dumoulin, M. et al. 2003, Nature 424: 783-788; Pleschberger, M. et al. 2003, Bioconjugate Chem 14: 440-448; Cortez-Retamozo, V. et al. 2002, Int J Cancer 89: 456-62; and Lauwereys, M. et al. 1998, EMBO J 17: 3512-3520. Engineered libraries of camelid antibodies and antibody fragments are commercially available, for example, from , Ghent, Belgium. As with other antibodies of non-human origin, an amino acid sequence of a camelid antibody can be altered recombinantly to obtain a sequence that more closely W0 2014/122613 resembles a human ce, i.e., the nanobody can be "humanized". Thus the natural low antigenicity of came|id antibodies to humans can be further reduced.

The came|id nanobody has a molecular weight approximately one-tenth that of a human lgG molecule and the protein has a physical er of only a few nanometers. One consequence of the small size is the ability of d nanobodies to bind to antigenic sites that are functionally invisible to larger antibody proteins, i.e., came|id nanobodies are useful as reagents detecting antigens that are otherwise cryptic using classical immunological techniques, and as possible eutic agents. Thus yet another consequence of small size is that a came|id nanobody can inhibit as a result of binding 1O to a specific site in a groove or narrow cleft of a target protein, and hence can serve in a capacity that more closely resembles the on of a classical low molecular weight drug than that of a classical antibody.

The low molecular weight and compact size further result in d nanobodies being extremely thermostable, stable to extreme pH and to proteolytic ion, and poorly antigenic. r consequence is that came|id dies readily move from the circulatory system into tissues, and even cross the blood-brain barrier and can treat disorders that affect nervous tissue. Nanobodies can further facilitated drug transport across the blood brain barrier. See US. Patent Publication No. 20040161738 published August 19, 2004. These features ed with the low antigenicity to humans indicate great eutic potential. Further, these molecules can be fully expressed in prokaryotic cells such as E. coli and are expressed as fusion proteins in bacteriophage and are functional.

Engineered nanobodies can further be customized by genetic engineering to have a half-life in a recipient subject of from 45 minutes to two weeks. In a specific ment, the came|id antibody or nanobody is obtained by grafting the CDRs sequences of the heavy or light chain of one of the human antibodies of the disclosure, XABl, XABZ, XABS, XAB4 or XABS, into nanobody or single domain antibody framework sequences, as described for example in PCT Publication No. WO 94/04678.

Non-antibody scaffold Known non-immunoglobulin frameworks or scaffolds include, but are not limited to, Adnectins (fibronectin) und Therapeutics, Inc, Waltham, MA), ankyrin (Molecular Partners AG, Zurich, Switzerland), domain antibodies (Domantis, Ltd idge, MA) and Ablynx nv (Zwijnaarde, Belgium)), lipocalin (Anticalin) (Pieris Proteolab AG, Freising, Germany), small modular immuno-pharmaceuticals (Trubion Pharmaceuticals Inc, Seattle, WA), maxybodies (Avidia, Inc. (Mountain View, CA)), Protein A ody AG, Sweden) and n (gamma-crystallin or ubiquitin) (Scil Proteins GmbH, Halle, Germany), protein epitope mimetics (Polyphor Ltd, Allschwil, rland). (a) Fibronectin scaffold The fibronectin lds are based preferably on ectin type III domain (e.g., the tenth module of the fibronectin type III (10 Fn3 domain)). The fibronectin type III domain has 7 or 8 beta strands which are distributed between two beta sheets, which themselves pack against each other to form the core of the protein, and further containing loops gous to CDRs) which connect the beta strands to each other and are solvent exposed. There are at least three such loops at each edge of the beta sheet sandwich, where the edge is the boundary of the protein perpendicular to the direction of the beta strands (US Patent No. 6,818,418).

These fibronectin-based scaffolds are not an immunoglobulin, although the overall fold is closely related to that of the smallest functional antibody nt, the variable region of the heavy chain, which comprises the entire antigen recognition unit in camel and llama lgG. Because of this structure, the non-immunoglobulin antibody mimics antigen binding ties that are similar in nature and affinity to those of antibodies. These scaffolds can be used in a loop randomization and shuffling strategy in vitro that is r to the process of affinity maturation of antibodies in vivo. These fibronectin- based molecules can be used as scaffolds where the loop regions of the molecule can be replaced with CDRs of one of XABl, XABZ, XABS, XAB4 or XAB5 using standard cloning techniques.

W0 2014/122613 (b) Ankyrin — Molecular Partners The technology is based on using proteins with ankyrin derived repeat modules as scaffolds for bearing variable regions which can be used for binding to different targets.

The ankyrin repeat module is a 33 amino acid polypeptide consisting of two anti-parallel ces and a B-turn. Binding of the variable regions is mostly zed by using ribosome display. (c) Maxybodies/AVimers - Avidia Avimers are derived from natural A-domain containing protein such as LRP-1. These domains are used by nature for protein-protein interactions and in human over 250 1O proteins are structurally based on A-domains. Avimers consist of a number of different "A-domain" monomers (2-10) linked via amino acid linkers. Avimers can be created that can bind to the target antigen using the methodology described in, for example, US Patent Publication Nos 20040175756; 20050053973; 20050048512; and 20060008844. (d) n A — Affibody Affibody® affinity ligands are small, simple proteins composed of a three-helix bundle based on the scaffold of one of the lgG-binding domains of Protein A. n A is a e protein from the ium lococcus aureus. This scaffold domain consists of 58 amino acids, 13 of which are randomized to generate Affibody® libraries with a large number of ligand variants (See e.g., US Patent No. 012). Affibody® molecules mimic antibodies; they have a molecular weight of 6 kDa, compared to the molecular weight of antibodies, which is 150 kDa. In spite of its small size, the binding site of Affibody® molecules is r to that of an antibody. (9) Anticalins — Pieris Anticalins® are ts developed by the company Pieris ProteoLab AG. They are d from lipocalins, a widespread group of small and robust proteins that are usually involved in the physiological transport or storage of chemically ive or insoluble compounds. Several natural lipocalins occur in human tissues or body liquids.

W0 2014/122613 The protein architecture is reminiscent of immunoglobulins, with hypervariable loops on top of a rigid framework. However, in contrast with dies or their recombinant fragments, lipocalins are composed of a single polypeptide chain with 160 to 180 amino acid residues, being just marginally bigger than a single immunoglobulin domain.

The set of four loops, which makes up the binding pocket, shows pronounced structural plasticity and tolerates a variety of side chains. The binding site can thus be reshaped in a proprietary process in order to recognize prescribed target molecules of different shape with high affinity and specificity.

One protein of lipocalin family, the bilin-binding n (BBP) of Pieris Brassicae has 1O been used to develop lins by mutagenizing the set of four loops. One example of a patent application describing "anticalins" is PCT Publication WO 873. (0 Affilin — Scil Proteins nTM molecules are small non-immunoglobulin proteins which are designed for specific affinities towards proteins and small molecules. New AffilinTM molecules can be very quickly selected from two libraries, each of which is based on a different human derived scaffold protein.

AffilinTM molecules do not show any ural homology to immunoglobulin proteins.

Scil Proteins employs two AffilinTM scaffolds, one of which is gamma crystalline, a human structural eye lens protein and the other is "ubiquitin" superfamily proteins. Both human scaffolds are very small, show high temperature stability and are almost resistant to pH changes and denaturing agents. This high stability is mainly due to the expanded beta sheet structure of the proteins. Examples of gamma crystalline derived proteins are described in W0200104144 and examples of itin-like" proteins are described in W02004106368. (g) Protein Epitope cs (PEM) PEM are medium-sized, cyclic, peptide-like molecules (MW 1-2kDa) mimicking betahairpin ary ures of ns, the major secondary structure involved in protein-protein interactions.

W0 2014/122613 Framework or Fc ering Engineered antibodies and proteins comprising an antigen-binding portion f of the sure include those in which modifications have been made to framework residues within VH and/or VL, e.g. to improve the properties of the antibody. Typically such framework modifications are made to decrease the immunogenicity of the antibody. For example, one approach is to utate" one or more framework residues to the ponding germline sequence. More ically, an antibody that has undergone c mutation may contain framework residues that differ from the germline sequence from which the antibody is derived. Such es can be identified by 1O comparing the antibody framework sequences to the germline sequences from which the antibody is derived. To return the framework region sequences to their germline configuration, the somatic mutations can be "backmutated" to the germline sequence by, for example, site-directed mutagenesis or PCR-mediated mutagenesis. Such "backmutated" antibodies are also intended to be encompassed by the disclosure.

Another type of ork modification involves mutating one or more residues within the framework region, or even within one or more CDR regions, to remove T cell - epitopes to thereby reduce the potential immunogenicity of the antibody. This approach is also referred to as "deimmunization" and is described in r detail in US. Patent Publication No. 20030153043 by Carr et al. 2O In addition or alternative to modifications made within the framework or CDR regions, antibodies of the disclosure may be ered to include modifications within the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and/or antigen-dependent cellular cytotoxicity. Furthermore, an antibody of the sure may be ally ed (e.g., one or more chemical moieties can be attached to the antibody) or be modified to alter its glycosylation, again to alter one or more functional properties of the antibody. Each of these embodiments is described in further detail below.

As used herein, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc region and variant Fc regions. The human lgG heavy chain Fc region is generally defined as comprising the amino acid residue from position C226 or from P230 to the carboxyl-terminus of the lgG W0 22613 2014/058854 antibody. The numbering of residues in the Fc region is that of the EU index of Kabat.

The C-terminal lysine (residue K447) of the Fc region may be removed, for example, during production or purification of the antibody. Accordingly, a composition of antibodies of the disclosure may comprise antibody tions with all K447 residues removed, antibody populations with no K447 residues removed, and antibody populations having a mixture of antibodies with and without the K447 residue.

In one embodiment, the hinge region of CH1 is modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased. This approach is described further in US. Patent No. 5,677,425 by Bodmer et al. The 1O number of cysteine es in the hinge region of CH1 is d to, for example, facilitate assembly of the light and heavy chains or to increase or decrease the ity of the antibody.

In another embodiment, the Fc hinge region of an antibody is mutated to decrease the biological half-life of the antibody. More ically, one or more amino acid mutations are introduced into the 3 domain interface region of the Fc-hinge fragment such that the antibody has impaired Staphylococcyl protein A (SpA) binding relative to native Fc-hinge domain SpA binding. This approach is described in further detail in US. Patent No. 745 by Ward et al.

In another embodiment, the antibody or protein comprising an antigen-binding portion thereof is ed to increase its biological half-life. Various approaches are possible.

For example, one or more of the following mutations can be introduced: T252L, T254S, T256F, as described in US. Patent No. 6,277,375 to Ward. Alternatively, to increase the biological half-life, the antibody can be altered within the CH1 or CL region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an lgG, as described in US. Patent Nos. 5,869,046 and 6,121,022 by Presta et al.

In yet other embodiments, the Fc region is altered by replacing at least one amino acid e with a different amino acid residue to alter the effector functions of the antibody or protein comprising an antigen-binding portion thereof. For example, one or more amino acids can be ed with a different amino acid residue such that the antibody has an altered affinity for an effector ligand but retains the antigen-binding ability of the W0 2014/122613 parent dy. The effector ligand to which affinity is d can be, for example, an Fc receptor or the C1 component of ment. This approach is described in further detail in US. Patent Nos. 5,624,821 and 5,648,260, both by Winter et al.

In another embodiment, one or more amino acids selected from amino acid es can be replaced with a different amino acid residue such that the antibody has altered C1q binding and/or reduced or abolished complement dependent cytotoxicity (CDC).

This approach is described in further detail in US. Patent Nos. 6,194,551 by ldusogie et In another embodiment, one or more amino acid residues are altered to thereby alter 1O the ability of the antibody to fix ment. This approach is described further in PCT Publication WO 94/29351 by Bodmer et at.

In yet another embodiment, the Fc region is modified to increase the ability of the antibody or protein comprising an n-binding portion thereof to mediate antibody dependent cellular xicity (ADCC) and/or to increase the affinity of the antibody for an Fcy receptor by modifying one or more amino acids. This approach is described further in PCT Publication WO 00/42072 by Presta. Moreover, the binding sites on human lgG1 for Fcle, Fclel, Fclell and FcRn have been mapped and ts with improved binding have been described (see Shields, R.L. et al. 2001, J. Biol. Chem 276:6591-6604).

In certain embodiments, the Fc domain of the lgG1 isotype is used. In some specific embodiments, a mutant variant of the lgG1 Fc fragment is used, e.g. a silent lgG1 Fc which reduces or ates the ability of the fusion polypeptide to mediate antibody dependent ar cytotoxicity (ADCC) and/or to bind to an Fcy receptor. An example of an lgG1 isotype silent mutant is lgG1 wherein Leucine is replaced by Alanine at amino acid positions 234 and 235 as described by Hezareh et al. 2001, J. Virol 75:12161-8 Another example of an lgG1 e silent mutant is lgG1 with D265A mutation (aspartate being substituted by alanine at position 265). In certain embodiments, the Fc domain is a silent Fc mutant preventing glycosylation at position 297 of the Fc domain.

For example, the Fc domain contains an amino acid tution of asparagine at position 297. An example of such amino acid substitution is the replacement of N297 by a glycine or an alanine.

W0 2014/122613 In still another embodiment, the glycosylation of an dy or protein comprising an antigen-binding portion thereof is modified. For example, an aglycoslated dy can be made (i.e., the dy lacks glycosylation). Glycosylation can be altered to, for example, increase the affinity of the antibody for the antigen. Such carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the dy ce. For example, one or more amino acid tutions can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site. Such aglycosylation may increase the ty of the antibody for antigen. Such an approach is 1O bed in further detail in US. Patent Nos. 5,714,350 and 6,350,861 by Co et al.

Additionally or alternatively, an dy or protein comprising an antigen-binding portion thereof can be made that has an d type of glycosylation, such as a cosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNac structures. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies. Such carbohydrate modifications can be accomplished by, for e, expressing the antibody in a host cell with altered glycosylation machinery. Cells with altered ylation machinery have been described in the art and can be used as host cells in which to s recombinant antibodies of the disclosure to thereby produce an antibody with altered glycosylation. For example, EP 1 176 195 by Hang et al. describes a cell line with a functionally disrupted FUT8 gene, which encodes a fucosyl transferase, such that antibodies expressed in such a cell line exhibit hypofucosylation. Therefore, in one embodiment, the antibodies of the disclosure are produced by recombinant expression in a cell line which exhibits a hypofucosylation pattern, for example, a mammalian cell line with deficient expression of the FUT8 gene encoding fucosyltransferase. PCT Publication WO 03/035835 by Presta describes a variant CHO cell line, Lecl3 cells, with reduced ability to attach fucose to Asn(297)-linked carbohydrates, also resulting in hypofucosylation of antibodies expressed in that host cell (see also Shields, R.L. et al. 2002, J. Biol. Chem. 277:26733-26740). PCT Publication WO 99/54342 by Umana et al. describes cell lines engineered to express glycoprotein-modifying glycosyl transferases (e.g., beta(1,4)-N acetylglucosaminyltransferase lll l)) such that antibodies expressed in the engineered cell lines exhibit increased bisecting GlcNac structures which results in sed ADCC activity of the antibodies (see also Umana et al. 1999, W0 2014/122613 Nat. Biotech. 17:176-180). atively, the antibodies and proteins comprising an antigen-binding portion thereof of the disclosure can be produced in a yeast or a filamentous fungus engineered for mammalian-like g|ycosy|ation pattern, and capable of producing dies lacking fucose as g|ycosy|ation pattern (see for example EP 1 297 172). r modification of the antibodies and proteins comprising an antigen-binding portion f as disclosed herein that is contemplated by the disclosure is pegylation.

These molecules can be pegylated to, for example, increase their biological (e.g., serum) half-life. For example, to pegylate an antibody, the antibody, or fragment thereof, 1O typically is reacted with polyethylene glycol (PEG), such as a reactive ester or de tive of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody fragment. The pegylation can be carried out by an acylation reaction or an tion reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any of the forms of PEG that have been used to derivatize other ns, such as mono 0) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody to be pegylated is an aglycosylated antibody. Methods for pegylating proteins are known in the art and can be applied to the antibodies of the disclosure. See for example, EP 0 154 316 by Nishimura 2O et al. and EP 0 401 384 by lshikawa et al.

Another modification of the antibodies and proteins comprising an n-binding portion thereof that is contemplated by the disclosure is a conjugate or a protein fusion of at least the antigen-binding region of the antibody of the disclosure to serum n, such as human serum albumin or a fragment thereof to increase half-life of the resulting molecule. Such approach is for example described in Ballance et al. EP 0 322 094.

Another possibility is a fusion of at least the antigen-binding region of the antibody of the disclosure to proteins capable of binding to serum proteins, such human serum albumin to increase ife of the resulting molecule. Such approach is for e described in Nygren et al., EP 0 486 525.

W0 2014/122613 Methods of engineering altered antibodies As discussed above, the anti-lL-17A antibodies having VH and VL sequences or full length heavy and light chain sequences shown herein can be used to create new anti- lL-17A antibodies by modifying full length heavy chain and/or light chain sequences, VH and/or VL sequences, or the constant region(s) attached thereto. Thus, in another aspect of the disclosure, the structural features of an anti-lL-17A antibody of the disclosure are used to create structurally related anti-lL-17A antibodies or protein comprising an antigen-binding portion thereof that retain at least one functional property of the antibodies disclosed herein, such as binding to human lL-17A and also inhibiting one or more functional properties of lL-17A (e.g., inhibiting binding of lL-17A or lL-17AF to its receptor(s), inhibiting lL-17A or lL-17AF induced tion of lL-6, GRO-alpha, etc.) inhibitory activity in in vivo assays.

Other antibodies ing substantially the same binding properties to lL-17A include ic dies or CDR grafted antibodies of any one of XABl, XABZ, XABS, XAB4 or XAB5 which retain the same VH and VL regions, or the CDR regions, of any one of XABl, XABZ, XABS, XAB4 or XAB5 and different constant regions or framework regions (for example a different Fc region selected from a different isotype, for example lgG4 or lgG2).

For example, one or more CDR regions of any one of XABl, XABZ, XABS, XAB4 or XABS, or mutations thereof, can be combined recombinantly with known framework regions and/or other CDRs to create additional, recombinantly—engineered, anti-lL-17A dies of the disclosure, as discussed above. Other types of modifications include those bed in the previous section. The starting al for the engineering method is one or more of the VH and/or VL sequences of XABl, XABZ, XABS, XAB4 or XAB5 provided in the tables above, or one or more CDR region thereof. To create the engineered antibody, it is not necessary to actually prepare (i.e., express as a protein) an antibody having one or more of the VH and/or VL sequences of XABl, XABZ, XABS, XAB4 or XABS, or one or more CDR regions thereof. Rather, the ation contained in the sequence(s) is used as the starting al to create a "second generation" ce(s) derived from the al sequence(s) and then the "second generation" sequence(s) is prepared and expressed as a protein.

W0 2014/122613 The second generation sequences are derived for example by altering the DNA coding sequence of at least one amino acid residue within the heavy chain variable region antibody sequence and/or the light chain variable region antibody sequence of any one of XABl, XAB2, XABS, XAB4 or XAB5, to create at least one altered antibody sequence; and expressing the altered antibody sequence as a protein.

Accordingly, in another embodiment, the disclosure es a method for preparing an anti-lL-17A antibody optimized for expression in a mammalian cell consisting of: a full length heavy chain antibody sequence a full length light chain antibody ce of any one of XABl, XAB2, XABS, XAB4 or XAB5; altering at least one codon in the tide 1O coding sequence, said codon encoding an amino acid residue within the full length heavy chain antibody sequence and/or the full length light chain antibody sequence to create at least one altered antibody sequence; and expressing the altered dy sequence as a protein.

The d antibody sequence can also be prepared by screening antibody libraries having unique heavy and light CDR3 sequences of any one of XABl, XAB2, XABS, XAB4 or XAB5 tively, or minimal essential binding inants as described in US Patent Publication No. 20050255552, and ative sequences for CDR1 and CDR2 sequences. The screening can be performed according to any screening technology appropriate for screening antibodies from antibody libraries, such as phage display technology.

Standard molecular biology techniques can be used to prepare and express the altered antibody sequence. The antibody encoded by the altered antibody sequence(s) is one that retains one, some or all of the desired functional properties of the anti-lL-17A antibodies bed herein, which functional properties include, but are not limited to, specifically g to human lL-17A; and/or it inhibits binding of lL-17A to its receptor(s); and/or it inhibits lL-17A induced production of, for example, lL-6 or GRO- alpha etc.

The d antibody may t one or more, two or more, or three or more of the functional properties discussed above.

W0 22613 In certain embodiments of the methods of ering antibodies of the disclosure, mutations can be introduced randomly or selectively along all or part of an anti-lL-17A antibody coding sequence and the resulting modified anti-lL-17A antibodies can be ed for binding activity and/or other functional ties as described herein.

Mutational methods have been described in the art. For example, PCT Publication WO 02/092780 by Short describes methods for ng and screening antibody mutations using saturation mutagenesis, synthetic ligation assembly, or a combination thereof. Alternatively, PCT Publication WO 03/074679 by Lazar et al. describes methods of using ational screening methods to optimize physiochemical properties of antibodies.

Nucleic acid molecules encoding antibodies of the disclosure r aspect of the disclosure pertains to nucleic acid molecules that encode the antibodies or proteins of the disclosure. Examples of variable light chain nucleotide sequences are those encoding the variable light chain amino acid sequences of any one of XABl, XAB2, XABS, XAB4 and XABS, the latter sequences being d from the Table 3 (showing the entire tide coding sequences of heavy and light chains of XABl, XAB2, XABS, XAB4 or XAB5) and Table 2 (showing the amino acid sequences of the variable regions of XABl, XAB2, XABS, XAB4 or XAB5).

The disclosure also pertains to nucleic acid molecules that derive from the latter sequences having been optimized for protein expression in mammalian cells, for example, CHO cell lines.

The nucleic acids may be present in whole cells, in a cell lysate, or may be nucleic acids in a partially purified or substantially pure form. A c acid is "isolated" or "rendered substantially pure" when purified away from other cellular components or other contaminants, e.g., other cellular nucleic acids or proteins, by standard techniques, including alkaline/SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis and others well known in the art. See, F. Ausubel, et al., ed. 1987, Current Protocols in Molecular Biology, Greene hing and Wiley lnterscience, New York. A c acid of the sure can be, for example, DNA or RNA and may or may not contain intronic sequences. In an embodiment, the nucleic acid is a cDNA W0 2014/122613 molecule. The nucleic acid may be present in a vector such as a phage display vector, or in a recombinant plasmid .

Nucleic acids of the sure can be obtained using standard molecular y techniques. Once DNA fragments encoding, for example, VH and VL segments are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example to convert the variable region genes to full-length antibody chain genes, to Fab fragment genes or to an scFv gene. In these manipulations, a VL- or VH-encoding DNA fragment is ively linked to another DNA le, or to a fragment ng another protein, such as an antibody constant region or a flexible 1O linker. The term "operatively linked", as used in this context, is intended to mean that the two DNA fragments are joined in a functional manner, for example, such that the amino acid ces encoded by the two DNA fragments remain in-frame, or such that the protein is expressed under control of a desired er.

The isolated DNA encoding the VH region can be converted to a full-length heavy chain gene by operatively linking the VH-encoding DNA to another DNA molecule encoding heavy chain constant s (CH1, CH2 and CH3). The sequences of human heavy chain constant region genes are known in the art (see e.g., Kabat, E. A., el al. 1991, Sequences of Proteins of Immunological lnterest, Fifth Edition, US. Department of Health and Human Services, NlH ation No. 91-3242) and DNA fragments encompassing these regions can be obtained by standard PCR amplification. The heavy chain constant region can be an lgG1, lgG2, lgG3, lgG4, lgA, lgE, lgM or lgD constant region. In some embodiments, the heavy chain constant region is selected among lgG1 isotypes. For a Fab fragment heavy chain gene, the VH-encoding DNA can be operatively linked to another DNA molecule encoding only the heavy chain CH1 nt region.

The isolated DNA encoding the VL region can be converted to a full-length light chain gene (as well as to a Fab light chain gene) by operatively linking the VL-encoding DNA to another DNA molecule encoding the light chain constant , CL. The sequences of human light chain constant region genes are known in the art (see e.g., Kabat, E. A., et al. 1991, Sequences of Proteins of Immunological lnterest, Fifth Edition, US.

Department of Health and Human Services, NlH Publication No. 91-3242) and DNA W0 2014/122613 fragments encompassing these regions can be obtained by standard PCR amplification.

The light chain constant region can be a kappa or a lambda constant region.

To create an scFv gene, the VH- and VL-encoding DNA fragments are operatively linked to another fragment encoding a le linker, e.g., encoding the amino acid sequence (Gly4 -Ser)3, such that the VH and VL sequences can be expressed as a contiguous single-chain protein, with the VL and VH regions joined by the flexible linker (see e.g., Bird et al. 1988, Science 242:423-426; Huston et at. 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883; erty et al. 1990, Nature 348:552-554).

Isolation of recombinant antibodies of the sure 1O A variety of s of screening antibodies and proteins comprising an antigenbinding portion thereof have been described in the Art. Such methods may be divided into in vivo systems, such as transgenic mice capable of producing fully human dies upon n immunization and in vitro s, consisting of ting dy DNA coding libraries, expressing the DNA library in an riate system for antibody production, selecting the clone that express antibody candidate that binds to the target with the affinity selection criteria and recovering the corresponding coding sequence of the selected clone. These in vitro technologies are known as display logies, and include without limitation, phage display, RNA or DNA display, ribosome display, yeast or mammalian cell display. They have been well described in the Art (for a review see for example: Nelson et al. 2010, Nature Reviews Drug discovery, "Development trends for human monoclonal antibody therapeutics" (Advance Online Publication) and Hoogenboom et al. 2001, Method in Molecular Biology 178:1- 37, O’Brien et al., ed., Human Press, , N.J.). In one specific embodiment, human recombinant antibodies of the disclosure are isolated using phage display methods for screening libraries of human recombinant antibody libraries, such as HuCAL® libraries.

Repertoires of VH and VL genes or related CDR regions can be separately cloned by polymerase chain reaction (PCR) or synthesized by DNA synthesizer and recombined randomly in phage libraries, which can then be screened for antigen-binding .

Such phage display methods for isolating human antibodies are established in the art or described in the examples below. See for example: US. Patent Nos. 5,223,409; ,403,484; and 5,571,698 to Ladner et al.; US. Patent Nos. 5,427,908 and 5,580,717 to W0 2014/122613 Dower et al.; US. Patent Nos. 5,969,108 and 197 to McCafferty et al.; and US.

Patent Nos. 5,885,793; 6,521,404; 6,544,731; 6,555,313; 6,582,915 and 6,593,081 to Griffiths et al.

In a certain embodiment, human antibodies directed against |L-17A can be identified using transgenic or transchromosomic mice carrying parts of the human immune system rather than the mouse system. These transgenic and transchromosomic mice include mice referred to herein as HuMAb mice and KM mice, respectively, and are collectively referred to herein as "human lg mice." The HuMAb mouse® (Medarex, Inc.) contains human immunoglobulin gene miniloci that 1O encode un-rearranged human heavy (u and y) and K light chain immunoglobulin ces, together with targeted mutations that inactivate the endogenous u and K chain loci (see e.g., Lonberg, et al. 1994, Nature 6-859). ingly, the mice exhibit reduced expression of mouse IgM or K, and in response to immunization, the introduced human heavy and light chain transgenes undergo class switching and somatic mutation to generate high affinity human IgGK monoclonal (Lonberg, N. et al. 1994, supra; reviewed in g, N., 1994 Handbook of Experimental cology 113:49-101; g, N. and Huszar, D. 1995, Intern. Rev. lmmunol. 13:65-93, and Harding, F. and Lonberg, N. 1995, Ann. N. Y. Acad. Sci. 764:536-546). The preparation and use of HuMAb mice, and the genomic modifications carried by such mice, is r described in Taylor, L. et al. 1992, Nucleic Acids Research 7-6295; Chen, J. et at. 1993, International Immunology 5:647-656; Tuaillon et al. 1993, Proc. Natl. Acad.

Sci. USA 94:3720-3724; Choi et al. 1993, Nature Genetics 4:117-123; Chen, J. et al. 1993, EMBO J. 12: 821-830; Tuaillon et al. 1994, J. Immunol. 12-2920; Taylor, L. et al. 1994, International Immunology 579-591; and Fishwild, D. et al. 1996, Nature Biotechnology 14: 845-851. See further, US. Patent Nos. 5,545,806; 825; ,625,126; 5,633,425; 5,789,650; 5,877,397; 5,661,016; 5,814,318; 5,874,299; and ,770,429; all to Lonberg and Kay; US. Patent No. 5,545,807 to Surani et al.; PCT Publication Nos. WO 92103918, WO 93/12227, WO 94/25585, WO 97113852, WO 98/24884 and WO 99/45962, all to Lonberg and Kay; and PCT Publication No. WO 01/14424 to Korman et al.

In another embodiment, human antibodies of the sure can be raised using a mouse that carries human immunoglobulin ces on transgenes and W0 2014/122613 transchomosomes such as a mouse that carries a human heavy chain ene and a human light chain transchromosome. Such mice, referred to herein as "KM mice", are described in detail in PCT Publication WO 02/43478 to |shida et al.

Still further, alternative enic animal systems expressing human immunoglobulin genes are available in the art and can be used to raise anti-lL-17A antibodies of the disclosure. For example, an alternative transgenic system referred to as the Xenomouse from x, Inc. can be used. Such mice are described in, e.g., US.

Patent Nos. 5,939,598; 6,075,181; 6,114,598; 6, 150,584 and 6,162,963 to Kucherlapati et al. As will be appreciated by a person d in the art, several other mouse models 1O used, such as the TrianniTM may be mouse from Trianni, Inc, the VeloclmmuneTM mouse from Regeneron Pharmaceuticals, Inc, or the KymouseTM mouse from Kymab Limited.

Moreover, alternative transchromosomic animal systems expressing human immunoglobulin genes are available in the art and can be used to raise anti-lL-17A antibodies of the disclosure. For example, mice carrying both a human heavy chain transchromosome and a human light chain transchromosome, referred to as "TC mice" can be used; such mice are bed in Tomizuka et al. 2000, Proc. Natl. Acad. Sci.

USA 97:722-727.

Human monoclonal antibodies of the disclosure can also be prepared using SCID mice into which human immune cells have been reconstituted such that a human antibody response can be generated upon immunization. Such mice are described in, for example, US. Patent Nos. 5,476,996 and 5,698,767 to Wilson et al.

Generation of onal antibodies of the sure from the murine system Monoclonal antibodies (mAbs) can be produced by a variety of techniques, including conventional monoclonal antibody methodology e.g., the standard somatic cell hybridization technique of Kohler and Milstein 1975, Nature 256:495. Many techniques for producing monoclonal antibody can be ed e.g., viral or nic transformation of B lymphocytes.

An animal system for preparing hybridomas is the murine system. Hybridoma production in the mouse is a well-established procedure. Immunization protocols and W0 2014/122613 techniques for isolation of immunized splenocytes for fusion are known in the art.

Fusion rs (e.g., murine myeloma cells) and fusion procedures are also known.

Chimeric or humanized antibodies of the present sure can be prepared based on the sequence of a murine monoclonal antibody prepared as described above. DNA encoding the heavy and light chain globulins can be obtained from the murine hybridoma of interest and engineered to contain non-murine (e.g., human) immunoglobulin sequences using standard molecular biology techniques. For example, to create a chimeric antibody, the murine variable regions can be linked to human constant regions using methods known in the art (see e.g., US. Patent No. 4,816,567 to 1O Cabilly et al.). To create a humanized antibody, the murine CDR regions can be inserted into a human framework using methods known in the art. See e.g., US. Patent No. 539 to Winter, and US. Patent Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.

Generation of hybridomas producing human monoclonal antibodies To generate hybridomas producing human monoclonal dies of the sure, splenocytes and/or lymph node cells from immunized mice can be isolated and fused to an appropriate immortalized cell line, such as a mouse a cell line. The resulting omas can be screened for the production of n-specific or epitope-specific antibodies. For example, single cell suspensions of splenic lymphocytes from immunized mice can be fused to xth the number of P3X63-Ag8.653 nonsecreting mouse myeloma cells (ATCC, CRL 1580) with 50% PEG. Cells are plated at approximately 2 x 145 in flat bottom microtiter plates, followed by a two week incubation in selective medium containing 20% fetal Clone Serum, 18% "653" conditioned media, % origen (IGEN), 4mM L-glutamine, 1mM sodium pyruvate, 5mM HEPES, 0:055mM 2- mercaptoethanol, 50 units/ml penicillin, 50mg/ml streptomycin, l ycin and 1X HAT (Sigma; the HAT is added 24 hours after the ). After approximately two weeks, cells can be cultured in medium in which the HAT is replaced with HT.

Individual wells can then be screened by ELISA for human monoclonal lgM and lgG antibodies. Once extensive hybridoma growth occurs, medium can be observed usually after 10-14 days. The antibody secreting hybridomas can be replated, screened again, and if still positive for human lgG, the monoclonal antibodies can be subcloned once or W0 2014/122613 twice by limiting dilution. The stable subclones can then be cultured in vitro to te small amounts of antibody in tissue culture medium for characterization.

To purify human monoclonal antibodies, selected hybridomas can be grown in two-liter spinner-flasks for monoclonal antibody purification. Supernatants can be filtered and concentrated before affinity chromatography with protein A-sepharose (Pharmacia, Piscataway, N.J.). Eluted lgG can be checked by gel electrophoresis and high performance liquid chromatography to ensure purity. The buffer on can be exchanged into PBS, and the concentration can be determined by Ongo using 1.43 extinction coefficient. The onal antibodies can be aliquoted and stored at -80° C. 1O Generation of transfectomas producing monoclonal antibodies dies of the disclosure can be produced in a host cell transfectoma using, for e, a combination of recombinant DNA techniques and gene transfection methods as is well known in the art (e.g., Morrison, S. 1985, Science 229:1202).

For example, to express the antibodies, or antibody fragments f, DNAs encoding partial or full-length light and heavy chains can be obtained by standard molecular biology or biochemistry techniques (e.g., DNA chemical synthesis, PCR ication or cDNA cloning using a hybridoma that expresses the antibody of interest) and the DNAs can be inserted into expression vectors such that the genes are operatively linked to transcriptional and translational control sequences. In this context, the term "operatively linked" is intended to mean that an antibody gene is ligated into a vector such that transcriptional and translational control sequences within the vector serve their ed function of regulating the transcription and translation of the antibody gene. The expression vector and expression control ces are chosen to be compatible with the expression host cell used. The antibody light chain gene and the antibody heavy chain gene can be inserted into te vector or, more typically, both genes are inserted into the same expression vector. The antibody genes are ed into the expression vector by standard methods (e.g., ligation of mentary restriction sites on the antibody gene fragment and vector, or blunt end ligation if no restriction sites are present). The light and heavy chain variable regions of the dies described herein can be used to create ength antibody genes of any antibody isotype by inserting them into expression vectors already encoding heavy chain constant and light chain W0 2014/122613 constant regions of the d e such that the VH segment is operatively linked to the CH segment(s) within the vector and the VL segment is operatively linked to the CL segment within the vector. Additionally or alternatively, the recombinant expression vector can encode a signal peptide, also called leader sequence, which facilitates secretion of the dy chain from a host cell. The antibody chain gene can be cloned into the vector such that the signal peptide is linked in frame to the amino terminus of the antibody chain gene. The signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide from a non-immunoglobulin protein).

Examples of such signal peptides are found in Table 7, and examples of polynucleotide sequences coding for the signal peptides are found in Table 8.

Table 7. Signal peptides for heavy and light e chains.

Signal peptide Sequence ID no. Used for heavy or light peptide chain SEQ ID NO: 60 Light Table 8. Polynucleotide ces coding for the signal peptides.

Coding for Sequence ID no. Coding signal peptide signal peptide sequence for heavy or light no. chain SEQ ID NO: 62 Light SEQ ID NO: 65 Heavy W0 22613 In addition to the antibody chain genes, the recombinant expression vectors of the disclosure carry regulatory ces that control the sion of the antibody chain genes in a host cell. The term "regulatory sequence" is intended to include promoters, enhancers and other sion control ts (e.g., polyadenylation signals) that control the transcription or translation of the antibody chain genes. Such regulatory sequences are described, for e, in Goeddel 1990, Gene Expression Technology.

Methods in logy 185, Academic Press, San Diego, CA). It will be appreciated by those skilled in the art that the design of the sion vector, including the selection of regulatory sequences, may depend on such factors as the choice of the host cell to be 1O transformed, the level of expression of protein desired, etc. Regulatory sequences for mammalian host cell expression include viral elements that direct high levels of protein expression in mammalian cells, such as promoters and/or enhancers derived from galovirus (CMV), Simian Virus 40 (SV40), adenovirus (e.g., the adenovirus major late promoter (AdMLP)), and a. Alternatively, nonviral regulatory sequences may be used, such as the ubiquitin promoter or P-globin promoter. Still further, regulatory elements composed of sequences from different sources, such as the SRa promoter system, which contains sequences from the SV40 early promoter and the long terminal repeat of human T cell leukemia virus type 1 (Takebe, Y. et al. 1988, Mol. Cell. Biol. 8:466-472).

In addition to the antibody chain genes and regulatory sequences, the inant expression vectors of the disclosure may carry additional sequences, such as sequences that regulate replication of the vector in host cells (e.g., origins of replication) and selectable marker genes. The able marker gene facilitates selection of host cells into which the vector has been introduced (see, e.g., US. Patent Nos. 4,399,216, 4,634,665 and 5,179,017, all by Axel et al.). For example, typically the selectable marker gene confers resistance to drugs, such as G418, hygromycin or methotrexate, on a host cell into which the vector has been introduced. Selectable marker genes include the dihydrofolate reductase (DHFR) gene (for use in dhfr- host cells with methotrexate selection/amplification) and the neo gene (for G418 selection).

For sion of the light and heavy chains, standard techniques were applied to transfect a host cell with the expression vector(s) encoding the heavy and light chains.

The various forms of the term "transfection" are intended to encompass a wide variety W0 2014/122613 2014/058854 of techniques commonly used for the uction of exogenous DNA into a prokaryotic or eukaryotic host cell, e.g., oporation, calcium-phosphate precipitation, DEAE- dextran transfection and the like. It is tically possible to express the antibodies of the disclosure in either prokaryotic or eukaryotic host cells. Expression of antibodies in eukaryotic cells, for example mammalian host cells, yeast or filamentous fungi, is discussed because such eukaryotic cells, and in particular mammalian cells, are more likely than prokaryotic cells to assemble and secrete a ly folded and immunologically active antibody.

In one specific embodiment, a cloning or expression vector according to the disclosure 1O comprises either at least one of coding sequences from Table 3, operatively linked to suitable promoter ces. In another specific embodiment, a cloning or expression vector according to the disclosure comprises either at least one of coding sequences from Table 4, operatively linked to suitable promoter sequences.

Mammalian host cells for expressing the inant antibodies of the disclosure include Chinese Hamster Ovary (CHO cells) (including dhfr- CHO cells, described Urlaub and Chasin 1980, Proc. Natl. Acad. Sci. USA 77:4216-4220 used with a DH FR able marker, e.g., as described in R.J. Kaufman and PA Sharp 1982, Mol. Biol. 1-621), CHOK1 dhfr+ cell lines, NSO myeloma cells, COS cells and SP2 cells. In particular, for use with NSO myeloma cells, another expression system is the GS gene expression system shown in PCT Publications WO 62, WO 89/01036 and EP 0 338 841. In one embodiment, mammalian host cells for sing the recombinant antibodies of the sure include mammalian cell lines deficient for FUT8 gene expression, for example as described in US Patent No. 6,946,292.

When recombinant expression vectors encoding antibody genes are introduced into mammalian host cells, the antibodies are produced by culturing the host cells for a period of time sufficient to allow for expression of the antibody in the host cells or secretion of the antibody into the culture medium in which the host cells are grown.

Antibodies can be recovered from the culture medium using standard protein purification methods (See for example Abhinav et al. 2007, Journal of Chromatography 848:28-37).

W0 2014/122613 In one specific ment, the host cell of the disclosure is a host cell transfected with an expression vector having the coding sequences selected from the group consisting of SEQ ID NO: 18, 31, 51, 19, 28, 32, 38, 40, 46, 48, 52, 56, and 58, suitable for the expression of XAB1, XAB2, XAB3, XAB4 or XAB5 respectively, operatively linked to suitable promoter sequences.

The latter host cells may then be further cultured under suitable conditions for the expression and tion of an antibody of the disclosure selected from the group consisting of XAB1, XAB2, XAB3, XAB4 or XAB5 respectively. lmmunoconjugates 1O In another aspect, the present disclosure features an anti-lL-17A antibody of the sure, or a fragment thereof, conjugated to an active or eutic moiety, such as a cytotoxin, a drug (e.g., an immunosuppressant) or a oxin. Such conjugates are referred to herein as "immunoconjugates". This may be particularly preferred if lL-17A is expressed on the surface of Th1? cells (Brucklacher-Waldert et al. 2009, J lmmunol. 183:5494-501). lmmunoconjugates that e one or more cytotoxins are referred to as otoxins." A cytotoxin or cytotoxic agent includes any agent that is detrimental to (e.g., kills) cells. es include taxon, cytochalasin B, gramicidin D, um bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, stine, t. colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1 -dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs f. Therapeutic agents also include, for example, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6- thioguanine, cytarabine, 5-fluorouracil decarbazine), ablating agents (e.g., mechlorethamine, thioepa chloraxnbucil, meiphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (ll) (DDP) cisplatin, anthracyclines (e.g., ubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents (e.g., vincristine and vinblastine).

W0 2014/122613 Other examples of therapeutic cytotoxins that can be conjugated to an antibody of the disclosure include mycins, eamicins, maytansines and auristatins, and derivatives thereof. An example of a calicheamicin antibody conjugate is commercially available (MylotargTM; Wyeth-Ayerst).

Cytoxins can be conjugated to antibodies of the sure using linker technology available in the art. Examples of linker types that have been used to conjugate a cytotoxin to an dy include, but are not limited to, hydrazones, thioethers, esters, disulfides and peptide-containing linkers. A linker can be chosen that is, for example, susceptible to cleavage by low pH within the lysosomal compartment or susceptible to 1O cleavage by proteases, such as proteases preferentially expressed in tumor tissue such as cathepsins (e.g., cathepsins B, C, D).

For further discussion of types of cytotoxins, linkers and methods for conjugating therapeutic agents to antibodies, see also Saito, G. et al. 2003, Adv. Drug Deliv. Rev. 55:199-215; Trail, P.A. et al. 2003, Cancer lmmunol. lmmunother. -337; Payne, G. 2003, Cancer Cell 3:207-212; Allen, TM. 2002, Nat. Rev. Cancer 2:750-763; Pastan, l. and an, R. J. 2002, Curr. Opin. lnvestig. Drugs 31089-1091; Senter, PD. and er, C.J. 2001, Adv. Drug Deliv. Rev. 53:247-264.

Antibodies of the present disclosure also can be ated to a radioactive isotope to generate cytotoxic radiopharmaceuticals, also referred to as radioimmunoconjugates.

Examples of radioactive isotopes that can be conjugated to antibodies for use diagnostically or therapeutically include, but are not limited to, iodinem, indiumm, yttriumgo, and lutetiumm. Method for preparing radioimmunconjugates are ished in the art. Examples of radioimmunoconjugates are commercially available, ing ZevalinTM (DEC Pharmaceuticals) and BexxarTM (Corixa Pharmaceuticals), and similar methods can be used to prepare radioimmunoconjugates using the antibodies of the disclosure.

The antibody conjugates of the disclosure can be used to modify a given biological se, and the drug moiety is not to be construed as limited to classical al therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, an enzymatically active toxin, or active fragment thereof, such as abrin, ricin A, W0 22613 pseudomonas exotoxin, or diphtheria toxin; a n such as tumor necrosis factor or interferon-y; or, biological response modifiers such as, for example, kines, interleukin-1 ("lL1"), interleukin-2 ("lL2"), eukin-6 ("IL6"), granulocyte macrophage colony stimulating factor ("GM-CSF"), granulocyte colony stimulating factor ("G-CSF"), or other growth factors. ques for conjugating such therapeutic moiety to antibodies are well known, see, e.g., Amon et al.1985, "Monoclonal Antibodies For lmmunotargeting Of Drugs ln Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56; Hellstrom et at. 1987, odies For Drug Delivery", in Controlled Drug 1O Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 ; Thorpe 1985, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506; Thorpe et al. 1982, Immunol. Rev., 62:119-58. ific molecules In another aspect, the present disclosure features bispecific or multispecific molecules sing an anti-lL-17A/AF antibody or protein comprising an antigen-binding portion thereof of the disclosure. An antibody or protein of the disclosure can be derivatized or linked to another functional le, e.g., another peptide or protein (e.g., another antibody or ligand for a receptor) to generate a bispecific le that binds to at least two different binding sites or target molecules. The antibody or protein of the sure may in fact be derivatized or linked to more than one other functional molecule to generate multi-specific molecules that bind to more than two different binding sites and/or target molecules; such specific molecules are also intended to be encompassed by the term "bispecific molecule" as used . To create a bispecific molecule of the disclosure, an antibody or protein of the disclosure can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other binding molecules, such as another antibody, antibody fragment, peptide or binding c, such that a bispecific molecule results.

Accordingly, the present disclosure includes bispecific molecules comprising at least one first binding specificity for lL-17A, for example, one antigen-binding portion of any one of XAB1, XAB2, XAB3, XAB4 or XAB5 and a second binding specificity for a W0 2014/122613 second target epitope. For example, the second target epitope is another epitope of IL- 17A different from the first target epitope. Another example is a bispecific molecule sing at least one first binding specificity for lL-17A, for example, one antigen- binding portion of any one of XAB1, XAB2, XAB3, XAB4 or XAB5 and a second binding specificity for an epitope elsewhere within lL-17A or within r target antigen.

Additionally, for the disclosure in which the bispecific molecule is multi-specific, the molecule can further include a third g specificity, in addition to the first and second target epitope.

In one embodiment, the bispecific molecules of the sure comprise as a binding 1O specificity at least one antibody, or an antibody fragment thereof, including, e.g., an Fab, Fab', 2, Fv, or a single chain Fv. The antibody may also be a light chain or heavy chain dimer, or any minimal fragment thereof such as a Fv or a single chain construct as described in Ladner et al. US. Patent No. 4,946,778.

Other antibodies which can be employed in the bispecific molecules of the sure are murine, chimeric and humanized monoclonal antibodies.

The bispecific molecules of the present disclosure can be ed by conjugating the constituent binding specificities, using methods known in the art. For example, each g-specificity of the bispecific molecule can be generated separately and then conjugated to one another. When the binding specificities are proteins or peptides, a variety of coupling or cross-linking agents can be used for covalent conjugation.

Examples of cross-linking agents include protein A, iimide, N-succinimidyl-S- acetyl-thioacetate (SATA), 5,5'-dithiobis(2—nitrobenzoic acid) (DTNB), o- phenylenedimaleimide (oPDM), inimidyl(2—pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl) axane-l-carboxylate (sulfo-SMCC) (see e.g., Karpovsky et al. 1984, J. Exp. Med. 86; Liu, MA et al. 1985, Proc. Natl.

Acad. Sci. USA 828648). Other methods include those described in Paulus 1985, Behring lns. Mitt. No. 78,118-132; Brennan et al. 1985, Science 229:81-83), and Glennie et al. 1987, J. lmmunol. 139: 2367-2375). Conjugating agents are SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, IL).

W0 2014/122613 When the binding specificities are antibodies, they can be conjugated by dryl bonding of the C-terminus hinge regions of the two heavy chains. In a particular embodiment, the hinge region is modified to contain an odd number of sulfhydryl residues, for example one, prior to conjugation.

Alternatively, both binding specificities can be d in the same vector and expressed and assembled in the same host cell. This method is particularly useful where the bispecific molecule is a mAb x mAb, mAb x Fab, Fab x F(ab')2 or ligand x Fab fusion protein. A bispecific molecule of the disclosure can be a single chain molecule comprising one single chain antibody and a binding determinant, or a single chain 1O bispecific molecule comprising two binding inants. Bispecific molecules may comprise at least two single chain molecules. Methods for preparing bispecific molecules are described for e in US. Patent Number 5,260,203; US. Patent Number 5,455,030; US. Patent Number 4,881,175; US. Patent Number 5,132,405; US. Patent Number 5,091,513; US. Patent Number 5,476,786; US. Patent Number 5,013,653; US. Patent Number 5,258,498; and US. Patent Number 5,482,858.

Binding of the bispecific molecules to their ic targets can be confirmed by, for e, enzyme-linked immunosorbent assay (ELISA), mmunoassay (REA), FACS analysis, bioassay (e.g., growth inhibition), or Western Blot assay. Each of these assays generally detects the presence of protein-antibody complexes of particular interest by employing a labeled reagent (e.g., an antibody) specific for the complex of interest.

Multivalent dies In another aspect, the present disclosure es multivalent antibodies comprising at least two cal or different antigen-binding portions of the antibodies of the disclosure g to lL-17A, for example, selected from antigen-binding portions of any one of XAB1, XAB2, XAB3, XAB4 or XAB5. In one embodiment, the multivalent antibodies provide at least two, three or four antigen-binding portions of the antibodies.

The antigen-binding portions can be linked together via protein fusion or nt or non-covalent linkage. Alternatively, methods of linkage have been described for the bispecific molecules. Tetravalent compounds can be obtained for example by cross- W0 2014/122613 linking dies of the disclosure with an antibody that binds to the constant regions of the antibodies of the disclosure, for example the Fc or hinge region.

Pharmaceutical compositions In another aspect, the present disclosure provides a composition, e.g., a pharmaceutical composition, containing one or a combination of antibodies or proteins comprising an antigen-binding portion thereof of the t disclosure, for example, one antibody selected from the group consisting of XABl, XABZ, XABS, XAB4 and XABS, formulated together with a pharmaceutically acceptable carrier. Such itions may e one or a combination of (e.g., two or more different) antibodies, or immunoconjugates or 1O ific molecules of the disclosure. For example, a ceutical composition of the disclosure can comprise a combination of antibodies or proteins that bind to different epitopes on the target n or that have complementary activities.

Pharmaceutical compositions of the disclosure also can be administered in combination therapy, i.e., combined with other agents. For example, the combination therapy can include an anti-lL-17A antibody or protein of the t disclosure, for example one antibody selected from the group ting of XABl, XABZ, XABS, XAB4 and XABS, combined with at least one other anti-inflammatory or another chemotherapeutic agent, for e, an immunosuppressant agent. Examples of therapeutic agents that can be used in combination therapy are described in greater detail below in the section on uses of the antibodies or proteins of the disclosure.

As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. The carrier should be suitable for intravenous, uscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). In one embodiment, the carrier should be suitable for subcutaneous route. Depending on the route of administration, the active nd, i.e., antibody, immunoconjugate, or bispecific molecule, may be coated in a material to protect the compound from the action of acids and other natural conditions that may vate the compound.

W0 2014/122613 The pharmaceutical compositions of the disclosure may include one or more pharmaceutically able salts. A "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound and does not impart any undesired toxicological effects (see e.g., Berge, S.M., et al. 1977, J. Pharm. Sci. 66:1-19). Examples of such salts include acid addition salts and base on salts.

Acid addition salts include those derived from ic inorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous and the like, as well as from nontoxic organic acids such as tic mono- and boxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, 1O aliphatic and aromatic sulfonic acids and the like. Base addition salts e those derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium and the like, as well as from nontoxic organic , such as N,N'- dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine and the like.

A pharmaceutical composition of the disclosure also may include a ceutically able xidant. Examples of pharmaceutically acceptable antioxidants include: water soluble antioxidants, such as ascorbic acid, ne hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; oil-soluble antioxidants, such as ascorbyl ate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Examples of suitable aqueous and eous carriers that may be employed in the pharmaceutical compositions of the disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the nance of the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of presence of microorganisms may be ensured both by sterilization procedures and by the inclusion of various W0 2014/122613 antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as, aluminum monostearate and n.

Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable ons or dispersion. The use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with 1O the active compound, use thereof in the pharmaceutical compositions of the disclosure is contemplated. Supplementary active compounds can also be incorporated into the compositions.

Therapeutic compositions typically must be e and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration.

The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper ty can be maintained, for e, by the use of a coating such as in, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, one can e isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. ged absorption of the injectable compositions can be brought about by ing in the ition an agent that delays absorption for example, monostearate salts and gelatin. s on the development of stable protein (e.g. antibody) formulations may be found in Cleland et al. 1993, Crit. Reviews. Ther. Drug Carrier Systems 10(4):307-377 and Wei Wang 1999, Int. J. Pharmaceutcs 9-88. Additional ation discussions for dies may be found, e.g., in Daugherty and Mrsny 2006, Advanced Drug Delivery Reviews 58: 686-706; US Pat. Nos 6,171,586, 4,618,486, US Publication No. 20060286103, PCT Publication WO 06/044908, WO 07/095337, WO 04/016286, Colandene et al. 2007, J. Pharm. Sci 96: 1598-1608; Schulman 2001, Am. J. Respir.

Crit. Care Med. 164:S6-S11 and other known references.

W0 22613 Solutions or suspensions used for intradermal or subcutaneous application typically include one or more of the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents, antibacterial agents such as benzyl alcohol or methyl parabens, antioxidants such as ascorbic acid or sodium bisulfite, chelating agents such ethylenediaminetetraacetic acid, buffers such as es, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. Such ations may be enclosed in ampoules, disposables syringes or multiple dose vials 1O made of glass or plastic.

Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as ed, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporating the antibodies or proteins of the sure into a sterile vehicle that contains a basic sion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

In one specific embodiment, the antibodies XABl, XAB2, XABS, XAB4 or XAB5 were administered as a liquid formulation in a vial. The amount of drug per vial was 150 mg.

The liquid contained 150 mg/mL antibody, 4.8 mM L-Histidine, 15.2 mM L-Histidine-HCI 220 mM Sucrose and 0.04% Polysorbate 20, at pH 6.0 1r 0.5. A 20% overfill was added to permit complete removal of the ed dose.

The amount of active ingredient which can be ed with a carrier material to produce a single dosage form will vary ing upon the subject being treated, and the particular mode of administration. The amount of active ient which can be ed with a carrier material to e a single dosage form will generally be that amount of the ition which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 0.01 per cent to about ninety-nine percent of active ingredient, from about 0.1 per cent to about 70 per cent, or from about W0 2014/122613 1 percent to about 30 percent of active ient in ation with a pharmaceutically acceptable carrier.

Dosage regimens are adjusted to provide the optimum desired response (e.g., a eutic response). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic ion. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and mity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary s for the subjects to be treated; each 1O unit contains a predetermined quantity of active compound ated to e the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the disclosure are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.

For administration of the antibody or protein, the dosage ranges from about 0.0001 to 150 mg/kg, such as 5, 15, and 50 mg/kg subcutaneous administration, and more usually 0.01 to 5 mg/kg, of the host body weight. For example dosages can be 0.3 mg/kg body weight, 1 mg/kg body weight, 3 mg/kg body weight, 5 mg/kg body weight or 10 mg/kg body weight or within the range of 1-10 mg/kg. An ary treatment regime entails administration once per week, once every two weeks, once every three weeks, once every four weeks, once per month, once every 3 months or once every three to 6 months. Dosage regimens for an anti-lL-17A antibody or protein of the disclosure include 1 mg/kg body weight, 3 mg/kg body weight, 5 mg/kg, 10 mg/kg, 20 mg/kg or 30 mg/kg by intravenous administration, with the antibody being given using one of the following dosing schedules: every four weeks for six dosages, then every three months; every three weeks; 3 mg/kg body weight once followed by 1 mg/kg body weight every three weeks.

In some methods, two or more antibodies with different binding specificities are administered simultaneously, in which case the dosage of each antibody administered falls within the ranges indicated. Antibodies or proteins of the sure are usually administered on multiple occasions. als between single dosages can be, for W0 2014/122613 example, weekly, monthly, every three months or yearly. lntervals can also be irregular as indicated by measuring blood levels of antibody to the target antigen in the patient. In some methods, dosage is ed to achieve a plasma dy concentration of about 1-1000 ug/ml and in some methods about 25-300 ug/ml.

Alternatively, antibody or protein can be administered as a sustained release formulation, in which case less nt administration is required. Dosage and frequency vary depending on the half-life of the antibody in the patient. In l, human antibodies show the longest ife, followed by humanized antibodies, chimeric antibodies, and nonhuman antibodies. The dosage and frequency of 1O administration can vary depending on whether the treatment is prophylactic or therapeutic. ln prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. Some ts continue to receive treatment for the rest of their lives. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes ed until progression of the e is reduced or terminated or until the patient shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regime.

Actual dosage levels of the active ingredients in the pharmaceutical itions of the present disclosure may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level will depend upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present disclosure employed, or the ester, salt or amide thereof, the route of stration, the time of administration, the rate of excretion of the particular compound being employed, the on of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

A peutically ive " of an anti-lL-17A antibody or protein of the disclosure can result in a decrease in severity of disease symptoms, an increase in W0 2014/122613 frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction.

A composition of the present disclosure can be administered by one or more routes of administration using one or more of a variety of s known in the art. As will be appreciated by the skilled n, the route and/or mode of administration will vary depending upon the desired results. Routes of administration for antibodies of the disclosure include intravenous, intramuscular, ermal, intraperitoneal, subcutaneous, spinal or other parenteral routes of administration, for example by ion or infusion. The phrase "parenteral administration" as used herein means 1O modes of administration other than enteral and topical administration, usually by injection, and es, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, pinal, epidural and intrastemal ion and infusion.

Alternatively, an dy or protein of the disclosure can be stered by a nonparenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, gually or topically.

The antibodies or proteins of the disclosure can be prepared with carriers that will protect the antibodies against rapid e, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems.

Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, ycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, e.g., Sustained and Controlled Release Drug ry Systems, J.R. Robinson, ed., Marcel Dekker, Inc, New York, 1978.

Therapeutic compositions can be administered with medical devices known in the art.

For example, in one embodiment, a therapeutic composition of the disclosure can be administered with a needleless hypodermic injection device, such as the s shown in US. Patent Nos. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824 or 4,596,556. Examples of well-known implants and modules useful in the present disclosure include: US. Patent No. 4,487,603, which shows an implantable micro- W0 2014/122613 infusion pump for dispensing medication at a controlled rate; US. Patent No. 4,486,194, which shows a therapeutic device for administering medicants through the skin; US.

Patent No. 4,447,233, which shows a medication infusion pump for delivering medication at a precise infusion rate; US. Patent No. 4,447,224, which shows a variable flow implantable on apparatus for continuous drug delivery; US. Patent No. 4,439,196, which shows an osmotic drug delivery system having multi-chamber compartments; and US. Patent No. 4,475,196, which shows an osmotic drug delivery system. Many other such implants, delivery systems, and s are known to those skilled in the art. 1O In certain embodiments, the antibodies or proteins of the disclosure can be formulated to ensure proper distribution in vivo. For example, the blood-brain r (BBB) excludes many highly hydrophilic compounds. To ensure that the therapeutic compounds of the disclosure cross the BBB (if desired), they can be formulated, for example, in liposomes. For methods of manufacturing liposomes, see, e.g., US.

Patents 4,522,811; 5,374,548; and 331. The liposomes may comprise one or more moieties which are selectively transported into ic cells or organs, thus enhance targeted drug delivery (see, e.g., V.V. Ranade 1989, J. Cline Pharmacol. 29:685). Exemplary targeting moieties e folate or biotin (see, e.g., US. Patent ,416,016 to Low et al.); mannosides (Umezawa et al. 1988, Biochem. Biophys. Res.

Commun. 153:1038); antibodies (P.G. Bloeman et al. 1995, FEBS Lett. 0; M.

Owais et al. 1995, Antimicrob. Agents ther. 39:180); surfactant n A receptor (Briscoe et al. 1995, Am. J. Physiol.1233:134); p120 (Schreier et al. 1994, J.

Biol. Chem. 269:9090); see also Keinanen and Laukkanen 1994, FEBS Lett. 346:123; Killion andFidler 1994, lmmunomethods 4:273.

Uses and methods of the disclosure The antibodies or proteins of the present sure have in vitro and in vivo stic and therapeutic utilities. For example, these les can be administered to cells in culture, e.g. in vitro or in vivo, or in a subject, e.g., in vivo, to treat, prevent or diagnose a variety of disorders.

W0 2014/122613 The methods are particularly suitable for treating, preventing or diagnosing lL-17A - related disorders and/or autoimmune and inflammatory disorders, e.g., rheumatoid tis, or psoriasis.

Specifically, the disclosure provides a method of treating lL-17A -re|ated disorders and/or autoimmune and inflammatory disorders. In certain embodiments the method comprises the step of stering isolated antibody or n comprising an antigen- binding portion thereof, according to the disclosure, to a subject in need thereof.

The disclosure also provides methods for decreasing or suppressing lL-17A or lL-17AF induced signaling response in target cells or tissues by ting a cell with a 1O ition comprising a therapeutically effective dose of the antibodies of the disclosure.

The disclosure also provides methods for decreasing levels of IL6, CXCL1, lL-8, GM- CSF and/or CCL2 in a cell comprising the step of contacting a cell with antibody or antigen binding fragment, such as a protein comprising an n-binding n thereof, according to the disclosure.

In the t description the phrase "lL-17A/AF mediated disease" or "lL-17A/AF - related disorder" encompasses all diseases and medical conditions in which lL-17A or lL-17AF plays a role, whether directly or indirectly, in the disease or medical condition, ing the causation, pment, progress, persistence or pathology of the disease or condition. Accordingly these terms include conditions associated with or characterized by aberrant lL-17A/AF levels and/or diseases or ions that can be treated by reducing or suppressing lL-17A/AF induced activity in target cells or s e.g. the production of lL-6 or pha. These include matory conditions and autoimmune diseases, such as arthritis, rheumatoid arthritis, or psoriasis. These further include allergies and allergic conditions, hypersensitivity reactions, chronic obstructive pulmonary disease, cystic fibrosis and organ or tissue transplant rejection.

For example, the antibodies or proteins of the disclosure may be used for the treatment of recipients of heart, lung, combined heart-lung, liver, kidney, pancreatic, skin or corneal lants, including allograft rejection or xenograft rejection, and for the W0 2014/122613 prevention of graft-versus-host disease, such as following bone marrow transplant, and organ transplant associated arteriosclerosis.

The antibodies or proteins of the disclosure, whilst not being limited to, are useful for the treatment, prevention, or amelioration of autoimmune disease and of inflammatory conditions, in particular inflammatory conditions with an aetiology including an autoimmune component such as arthritis (for example rheumatoid arthritis, arthritis chronica progrediente and arthritis deformans) and rheumatic diseases, including inflammatory conditions and rheumatic diseases involving bone loss, inflammatory pain, spondyloarhropathies including ankylosing spondylitis, Reiter syndrome, ve 1O arthritis, psoriatic arthritis, juvenile idiopathic arthritis and enterophathis tis, enthesitis, hypersensitivity (including both ainNays hypersensitivity and dermal ensitivity) and allergies. Specific auto-immune es for which antibodies of the disclosure may be employed include autoimmune haematological disorders (including e.g. hemolytic a, aplastic anaemia, pure red cell anaemia and thic thrombocytopenia), systemic lupus erythematosus (SLE), lupus tis, inflammatory muscle diseases (dermatomyosytis), periodontitis, polychondritis, derma, Wegener granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, Steven-Johnson syndrome, idiopathic sprue, autoimmune matory bowel disease (including e.g. ulcerative colitis, s disease and irritable bowel syndrome), endocrine ophthalmopathy, ’ disease, sarcoidosis, le sclerosis, systemic sclerosis, fibrotic diseases, primary biliary cirrhosis, juvenile diabetes (diabetes mellitus type I), uveitis, keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitial lung fibrosis, periprosthetic osteolysis, glomerulonephritis (with and t nephrotic syndrome, e.g. ing idiopathic nephrotic me or minimal change nephropathy), multiple a other types of tumors, inflammatory disease of skin and cornea, myositis, loosening of bone implants, metabolic disorders, (such as y, atherosclerosis and other cardiovascular diseases including d cardiomyopathy, myocarditis, diabetes mellitus type II, and dyslipidemia), and autoimmune thyroid diseases (including Hashimoto thyroiditis), small and medium vessel primary vasculitis, large vessel vasculitides including giant cell arteritis, hidradenitis suppurativa, neuromyelitis optica, Sjogren’s syndrome, Behcet’s e, atopic and contact dermatitis, bronchiolitis, inflammatory muscle es, autoimmune peripheral neurophaties, immunological renal, hepatic and thyroid diseases, W0 2014/122613 inflammation and thrombosis, autoinflammatory fever syndromes, hematological disorders, and bullous diseases of the skin and mucous membranes. Anatomically, uveitis can be anterior, intermediate, posterior, or pan- uveitis. It can be chronic or acute. The etiology of uveitis can be autoimmune or non- infectious, infectious, associated with systemic disease, or a white-dot syndrome.

The antibodies or proteins of the disclosure may also be useful for the treatment, prevention, or amelioration of asthma, bronchitis, bronchiolitis, thic interstitial pneumonias, pneumoconiosis, pulmonary emphysema, and other obstructive or inflammatory diseases of the airways. 1O The antibodies or proteins of the disclosure may also be useful for treating diseases of bone metabolism including osteoarthritis, osteoporosis and other inflammatory arthritis, and bone loss in general, including age-related bone loss, and in particular periodontal disease.

In addition, the antibodies or proteins of the disclosure may also be useful for treating chronic candidiasis and other chronic fungal diseases, as well as cations of infections with tes, and complications of smoking are considered to be promising avenues of treatment, as well as viral infection and complications of viral infection.

Inhibition of lL-17 and its receptor is among the most ing new modes of actions (MOA) for the treatment of chronic inflammatory es, with psoriasis being the most advanced indication among several diseases currently being d for lL-17 modulator drug development (Miossec P and Kolls JK. 2012, Nat Rev Drug Discov.10:763-76).

Several studies have unambiguously trated that blocking lL-17A in ts with moderate to severe plaque psoriasis is safe in the short term and induces very remarkable improvements (e.g. Hueber W, Patel DD, Dryja T, et al 2010, Sci Transl Med. ;2:52ra72). These findings exceeded ations and confirmed the esis that lL-17A is a key signaling molecule in the pathogenesis of psoriasis (Garber K. 2012,Nat Biotechnology 30:475—477).

Furthermore, in l animal models including the most common multiple sclerosis (MS) model mental autoimmune encephalomyelitis, lL-17 is pivotal in the inflammatory processes (Bettelli E, et al 2008, Nature; 453:1051-57, Wang HH, et al W0 2014/122613 2011, J Clin Neurosci; 18(10):1313-7, Matsushita T, et al 2013, PLoS One; 8(4):e64835). |L-17 effects are mainly lammatory, and synergize with other cytokines. |L-17 effects such as ion of chemokine production by epithelial cells, upregulation of interleukin (|L)-1b, tumor necrosis factor alpha (TNFa) and matrix metalloproteinase (MMP)-9 in macrophages, and induction of the secretion of IL 6, lL-8 and prostaglandin E2, fit well with many aspects of the MS pathology. There is also data arguing against a pivotal role of |L-17 in neuroinflammation, including transgenic overexpression models in mice (Haak S et al 2009, JCI; 119:61-69).

Asthma is a heterogeneous inflammatory e of the always that is manifested 1O clinically by symptoms of airflow obstruction that varies in severity either neously or in response to treatment. While asthma has been considered to be driven by T helper cell type 2 (Th2) cells and their products, recent data t that a Th2-high gene signature is present in the always of only ~50% of subjects with asthma (Woodruff PG et al 2009, Am J Respir Crit Care Med 180:388—95). philic inflammation is dominant in acute severe ; some individuals with asthma present with ent sputum neutrophilia and a poor clinical response to inhaled steroids; and sputum neutrophilia is prominent in asthmatic duals taking large doses of d and/or oral steroids (Wenzel 2012, Nature Med 18:716-25).

Increased levels of |L-17A that correlate with the severity of asthma have been reported in the circulation and always of individuals with asthma compared to healthy controls.

Pre-clinical s in mouse models of allergic ary inflammation have implicated a requirement for |L-17A and its receptor (lL-17RA) in neutrophilic ain/vay inflammation and steroid-resistant ain/vay hyper responsiveness. Thus, the properties of |L-17A in vitro, its presence in increased amounts in asthma, and the pre-clinical models of the disease support a role for |L-17A in neutrophilic and/or Th2-low forms of the disease that are poorly responsive to steroids (Cosmi L et al 2009, Am J Respir Crit Care Med 180:388—95).

Thus, the following list of conditions comprises particularly preferred targets for treatment with antibodies or proteins comprising an antigen-binding portion thereof according to the disclosure: Multiple sclerosis, psoriasis, asthma, systemic lupus erythematosus (SLE), and lupus nephritis.

W0 2014/122613 The antibodies or proteins of the disclosure may be administered as the sole active ingredient or in conjunction with, e.g. as an adjuvant to or in combination to, other drugs e.g. immunosuppressive or immunomodulating agents or other nflammatory agents or other cytotoxic or anti-cancer agents, e.g. for the treatment or prevention of diseases mentioned above. For example, the antibodies of the disclosure may be used in combination with DMARD, e.g. Gold salts, sulphasalazine, antimalarias, methotrexate, D-penicillamine, azathioprine, enolic acid, tacrolimus, sirolimus, minocycline, leflunomide, glucocorticoids; a calcineurin inhibitor, e.g. porin A or FK 506; a modulator of lymphocyte recirculation, e.g. FTY720 and FTY720 analogs; a mTOR 1O inhibitor, e.g. rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, CCl779, ABT578, AP23573 or TAFA-93; an ascomycin having immuno-suppressive properties, e.g. ABT-281, ASM981, etc.; corticosteroids; cyclophosphamide; azathioprine; leflunomide; mizoribine; myco-pheno-late mofetil; 15-deoxyspergualine or an immunosuppressive homologue, ue or derivative f; immunosuppressive monoclonal antibodies, e.g., monoclonal antibodies to leukocyte receptors, e.g., MHC, CD2, CD3, CD4, CD7, CD8, CD25, CD28, CD40. CD45, CD58, CD80, CD86 or their ligands; other immunomodulatory compounds, e.g. a recombinant binding le having at least a portion of the extracellular domain of CTLA4 or a mutant thereof, e.g. an at least extracellular portion of CTLA4 or a mutant f joined to a LA4 protein sequence, e.g. CTLA4lg (for ex. designated ATCC 68629) or a mutant thereof, e.g.

LEA29Y; adhesion molecule inhibitors, e.g. LFA-1 antagonists, lCAM-1 or -3 antagonists, VCAM-4 antagonists or VLA-4 antagonists; or a chemotherapeutic agent, e.g. paclitaxel, gemcitabine, cisplatinum, doxorubicin or 5-fluorouracil; anti TNF , e.g. monoclonal antibodies to TNF, e.g. infliximab, adalimumab, CDP870, or or constructs to TNF-RI or TNF-Rll, e.g. Etanercept, PEG-TNF-Rl; blockers of proinflammatory cytokines, |L1 blockers, e.g. Anakinra or |L1 trap, canakinumab, |L13 blockers, |L4 blockers, |L6 blockers, other |L17 blockers (such as numab, broadalumab, ixekizumab); chemokines blockers, e.g tors or activators of proteases, e.g. metalloproteases, anti-IL15 dies, anti-lL6 antibodies, anti-lL4 antibodies, L13 antibodies, anti-CD20 antibodies, NSAle, such as n or an anti-infectious agent (list not limited to the agent mentioned).

In accordance with the foregoing the present disclosure provides in a yet further aspect: W0 2014/122613 A method as defined above comprising co-administration, e.g. concomitantly or in ce, of a therapeutically effective amount of an anti-lL-17A antibody or protein comprising an antigen-binding portion thereof as disclosed herein, and at least one second drug substance, said second drug substance being a immuno-suppressive / immunomodulatory, anti-inflammatory chemotherapeutic or anti-infectious drug, e.g. as indicated above.

Or, a therapeutic combination, e.g. a kit, comprising of a therapeutically effective amount of a) an antibody or protein or protein comprising an antigen-binding portion thereof as disclosed herein, and b) at least one second substance selected from an 1O immuno-suppressive / immunomodulatory, anti-inflammatory chemotherapeutic or anti- infectious drug, e.g. as indicated above. The kit may comprise instructions for its administration.

Where the antibodies or proteins comprising an antigen-binding portion thereof as disclosed herein are stered in conjunction with other immuno-suppressive / immunomodulatory, anti-inflammatory chemotherapeutic or anti-infectious y, dosages of the co-administered combination compound will of course vary depending on the type of co-drug employed, e.g. whether it is a DMARD, anti-TNF, |L1 blocker or others, on the specific drug employed, on the condition being treated and so forth.

In one embodiment, the antibodies or proteins comprising an n-binding portion thereof can be used to detect levels of lL-17A, or levels of cells that n lL-17A. This can be achieved, for example, by contacting a sample (such as an in vitro sample) and a l sample with the anti-lL-17A antibody (or n) under conditions that allow for the formation of a x between the antibody and lL-17A. Any complexes formed between the antibody (or n) and lL-17A are ed and compared in the sample and the control. For example, standard detection methods, well known in the art, such as ELISA and flow cytometric , can be performed using the compositions of the sure.

Accordingly, in one aspect, the disclosure further provides methods for detecting the presence of lL-17A (e.g., human lL-17A antigen) in a sample, or measuring the amount of lL-17A, comprising contacting the sample, and a control sample, with an antibody or n of the disclosure, or an n-binding portion thereof, which specifically binds W0 2014/122613 to IL-17A, under conditions that allow for formation of a complex between the antibody or portion thereof and IL-17A. The formation of a complex is then detected, wherein a difference in x formation between the sample and the control sample is indicative of the presence of IL-17A in the sample.

Also within the scope of the disclosure are kits consisting of the compositions (e.g., antibodies, ns, human antibodies and bispecific molecules) of the disclosure and instructions for use. The kit can further contain a least one additional reagent, or one or more onal dies or proteins of the disclosure (e.g., an antibody having a complementary activity which binds to an epitope on the target antigen distinct from the 1O first antibody). Kits typically include a label indicating the intended use of the contents of the kit. The term label es any g, or recorded material supplied on or with the kit, or which othenNise accompanies the kit. The kit may further comprise tools for diagnosing whether a t belongs to a group that will respond to an anti-IL-17A dy treatment, as d above.

The disclosure having been fully bed is now further illustrated by the following examples and claims, which are illustrative and are not meant to be further limiting.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 provides the three-dimensional structure of the XA81 Fab. Figure 1A is a space-filling representation. Figure 18 is a n representation.

Figure 2 provides the three-dimensional structure of the XA81 Fv complex with human IL-17A. Figure 2A shows the two XA81 Fv fragments in space-filling representation and Figure 28 shows the two XA81 Fv fragments in cartoon representation.

Figure 3 shows graphs of ELISA results according to an example. The graph numbering corresponds to the candidate designation as follows: 1 is M8440; 2 is M8464; 3 is M8468; 4 is M8444; 5 is M8435; 6 is M8463; 7 is XA81. Figure 3A is a graph g the normalized signal versus the Fab Concentration (M). Figure 38 is a graph showing the normalized remaining signal versus the washing incubation time (hours). Figure 3C is a graph showing the normalized signal versus the Fab competitor concentration (M).

W0 2014/122613 Figure 4 provides the three-dimensional structure of the XABZ Fv complex with human lL-17A. Figure 4A shows the two XABZ Fv fragments in space-filling entation and Figure 4B shows the two XABZ Fv fragments are shown in cartoon representation.

Figure 5 provides the dimensional structure of the XABZ Fv complex with human lL-17A as a close-up view.

Figure 6 provides the three-dimensional structure of the XAB5 Fv complex with human lL-17A. Figure 6A shows the two XAB5 Fv fragments in space-filling representation and Figure 6B shows the two XAB5 Fv fragments in cartoon representation.

Figure 7 es the three-dimensional structure of the XAB5 Fv x with human 1O lL-17A, as a close-up view of the antibody L-CDR1.

Figure 8 provides the three-dimensional structure of the XAB4 Fv x with human lL-17A. Figure 8A shows the two XAB4 Fv fragments in space-filling representation Figure 8B shows the two XAB4 Fv fragments in cartoon representation.

Figure 9 provides the three-dimensional structure of the XAB4 Fv complex with human lL-17A as a close-up view of the antibody L-CDR1.

Figure 10 es the three-dimensional ure of the XAB4 Fv complex with human lL-17A as a close-up view of the antibody L-CDR2.

Figure 11 is a graph showing the therapeutic score for XAB4 in an experimental autoimmune encephalomyelitis (EAE) model.

Figure 12 is a graph showing the therapeutic weight change (%) of s in the EAE model.

Figure 13 is a graph showing the cumulative therapeutic scores in the EAE model.

Figures 14 and 15 are graphs showing comparisons of the therapeutic score pre- and post-treatment in the EAE model.

Figure 16 is a graph showing the prophylactic score for XAB4 in the EAE model.

W0 2014/122613 Figure 17 is a graph showing the prophylactic weight change (%) of animals in the EAE model.

Figure 18 is a graph showing the cumulative lactic scores in the EAE model.

Figure 19 is a graph showing the maximum prophylactic scores in the EAE model.

Figure 20 is a graph showing the EAE onset in the EAE model.

Figure 21 is a graph g antagonistic effect of XAB4 on lL-6 e in a human astrocyte model.

Figure 22 is a graph showing antagonistic effect of XAB4 on CXCL1 release in the human astrocyte model. 1O Figure 23 is a graph showing antagonistic effect of XAB4 on lL-8 release in the human yte model.

Figure 24 is a graph showing antagonistic effect of XAB4 on GM-CSF release in the human astrocyte model.

Figure 25 is a graph showing antagonistic effect of XAB4 on CCL2 release in the human astrocyte model.

Examples XA81 is a human lgG1/K monoclonal antibody. It was generated using standard molecular biological ques. In brief, the Medarex system was used. Mice were immunized with recombinant human lL-17A. Mice were ized by C02 inhalation and spleen cells were harvested and fused with a myeloma cell line using PEG 4000.

Fused cells were plated into wells with a feeder layer of peritoneal cells. Supernatants were taken from cultured cells and assayed for lL-17A reactive mAbs by ELISA. Clones positive for the production of lL-17A mAbs were selected and plated out.

The hybridoma responsible for the secretion of XA81 was identified for further characterization on the basis of initial promising antibody/antigen binding characteristics W0 2014/122613 such as binding affinity for IL-17A, ability to block IL-17A binding to its receptor, and ability to block IL-17A mediated biological effects in in vitro assays.

The amino acid sequence of XAB1 is SEQ ID NO: 14 (heavy chain) and SEQ ID NO: 15 (light chain). XAB1 was chosen for uent affinity maturation.

As a first step toward structure-guided affinity maturation, the crystal structure of the XAB1 Fab in the free state as well as the ponding Fv complex with human IL-17A were determined as described below. The analysis of the three-dimensional structure of the XAB1 Fv x with human IL-17A allowed for a rational affinity maturation s to be carried out alongside, and as an alternative to, a more randomised 1O process. Further details are ed below.

In addition, X-ray crystallography was used to characterise some of the affinity matured variant antibodies that were generated. Analysis of crystal data from the ty matured variants allowed for a deeper understanding of the binding behaviour of the variant antibodies and some unexpected properties were discovered as will be described further below.

Example 1. l ure of the XAB1 Fab in the free state (i) al and Methods Standard molecular biological protocols were used to obtain the XAB1 Fab antibody fragment. In brief, the Fab was cloned and expressed in E. co/iW311O with a C-terminal hexahistidine tag on the heavy-chain. The recombinant protein was purified by Ni- chelate chromatography followed by size-exclusion chromatography on a SPX-75 column in 10mM TRIS pH 7.4, 25mM NaCl. The XAB1 Fab was then concentrated by ultra-filtration to 10.4mg/m| and crystallized.

Standard crystallization protocols were followed. In brief, ls were grown at 19°C in SD2 96 well-plates, using the method of vapour diffusion in sitting drops. The protein stock was mixed 1:1 with a crystallization buffer containing 40% PEG 300, 0.1M sodium phosphate-citrate pH 4.2. Total drop size was 0.4 ul. Prior to X-ray data collection, one crystal was mounted in a nylon cryo-loop and directly flash cooled into liquid nitrogen.

W0 2014/122613 X-ray data collection and processing was carried out using standard ols. y, X-ray data to 2.1A resolution were collected at the Swiss Light , beamline X1OSA, with a MAR225 CCD detector, using 1.0000A X-ray radiation. In total, 180 images of 10° oscillation each were recorded at a crystal-to-detector distance of 190mm and processed with the HKL2000 re package. The crystal belonged to space group C2 with cell parameters a=51.63A, b=132.09A, c=77.25A, or = 90000, B = 98.880, y = 90.000 and one XAB1 Fab molecule in the asymmetric unit. R-sym to 2.1A resolution was 10.4% and data completeness 99.0%.

The structure was determined by molecular replacement with the program PHASER. 1O Search models for the VHNL and CH1/CL domains were ted from PDB entry 1HEZ. Iterative model ng and refinement were performed with the programs Coot (Crystallographic Object-Oriented Toolkit) and CNX (Crystallography & NMR eXplorer) version 2002, until no further significant improvements could be made to the model.

Final R- and R-free for all data were 0.188 and 0.231, respectively. The final refined model showed a root-mean-square deviation (RMSD) from ideal bond lengths and bond angles of 0.004A and 09°, tively. (ii) Results The results of the X-ray refinement of the XAB1 Fab are provided in Table 9 and the three-dimensional structure is shown in Figure 1.

Table 9. X-ray ment of the XAB1 Fab with the program CNX.

REMARK 3 REMARK 3REF|NEMENT.

REMARK 3 PROGRAM : CNX 2002 REMARK 3 AUTHORS :Brunger, Adams, Clore, Delano, REMARK 3 Gros, Grosse-Kunstleve, Jiang, REMARK 3 Kuszewski, Nilges, Pannu, Read, REMARK 3 Rice, Simonson, Warren REMARK 3 and REMARK 3 Accelrys Inc., REMARK 3 Yip, Dzakula).

REMARK 3 REMARK 3 DATA USED IN REFINEMENT.

REMARK 3 RESOLUTION RANGE HIGH ROMS) : 2.10 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS): 33.33 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.0 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 1964563062 REMARK 3 DATA CUTOFF LOW (ABS(F)): 0.000000 REMARK COMPLETENESS (WORKING+TEST) (%) : 98.2 REMARK NUMBER OF REFLECTIONS : 29298 REMARK REMARK FIT TO DATA USED IN REFINEMENT.

REMARK CROSS-VALIDATION METHOD : THROUGHOUT REMARK FREE R VALUE TEST SET SELECTION : RANDOM REMARK R VALUE (WORKING SET) : 0.188 REMARK FREE R VALUE : 0.231 REMARK FREE R VALUE TEST SET SIZE (%) : 4.9 REMARK FREE R VALUE TEST SET COUNT : 1436 REMARK ESTIMATED ERROR OF FREE R VALUE : 0.006 REMARK REMARK FIT IN THE HIGHEST RESOLUTION BIN.

REMARK TOTAL NUMBER OF BINS USED : 6 REMARK wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww BIN RESOLUTION RANGE HIGH (A) : 2.10 REMARK BIN RESOLUTION RANGE LOW (A) : 2.23 REMARK BIN COMPLETENESS (WORKING+TEST) (%) : 94.7 REMARK REFLECTIONS IN BIN (WORKING SET) : 4478 REMARK BIN R VALUE (WORKING SET) : 0.201 REMARK BIN FREE R VALUE : 0.241 REMARK BIN FREE R VALUE TEST SET SIZE (%) : 4.5 REMARK BIN FREE R VALUE TEST SET COUNT : 213 REMARK ESTIMATED ERROR OF BIN FREE R VALUE : 0.016 REMARK REMARK NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.

REMARK PROTEIN ATOMS : 3311 REMARK NUCLEIC ACID ATOMS : 0 REMARK HETEROGEN ATOMS : 5 REMARK SOLVENT ATOMS : 313 REMARK REMARK B .

REMARK FROM WILSON PLOT (A**2) : 21.1 REMARK MEAN B VALUE (OVERALL, A**2) : 27.4 REMARK OVERALL ROPIC B VALUE.

REMARK B11 (A**2) : -6.02 REMARK BZZ(A**2): 3.30 REMARK B33 (A**2): 2.73 REMARK B12(A**2): 0.00 REMARK B13(A**2): 3.82 REMARK BZ3(A**2): 0.00 REMARK REMARK BULK SOLVENT MODELING.

REMARK METHOD USED : FLAT MODEL REMARK KSOL : 0.399279 REMARK BSOL : 54.4727 (A**2) REMARK REMARK ESTIMATED COORDINATE ERROR.

REMARK ESD FROM LUZZATI PLOT (A) : 0.21 REMARK ESD FROM SIGMAA (A) : 0.12 REMARK LOW RESOLUTION CUTOFF (A) : 5.00 REMARK REMARK CROSS-VALIDATED ESTIMATED COORDINATE ERROR.

REMARK ESD FROM C-V I PLOT (A) : 0.29 REMARK ESD FROM C-V SIGMAA (A) : 0.14 REMARK REMARK RMS DEVIATIONS FROM IDEAL VALUES.

REMARK BOND LENGTHS (A) : 0.004 REMARK BOND ANGLES (DEGREES) : 0.9 REMARK AL ANGLES ES) : 21.4 REMARK IMPROPER ANGLES (DEGREES) : 0.58 REMARK 3 REMARK 3 ISOTROPIC THERMAL MODEL: RESTRAINED REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA REMARK 3 MAIN-CHAIN BOND (A**2): 1.41 ; 1.50 REMARK 3 MAIN-CHAIN ANGLE (A**2): 2.21 ; 2.00 REMARK 3 SIDE-CHAIN BOND (A**2): 2.31 ; 2.00 REMARK 3 SIDE-CHAIN ANGLE (A**2): 3.44; 2.50 REMARK 3 REMARK 3 NCS MODEL : NONE REMARK 3 REMARK 3 NCS RESTRAINTS. RMS VEIGHT REMARK 3 GROUP 1 POSITIONAL (A): NULL ;NULL REMARK 3 GROUP 1 B-FACTOR (A**2) : NULL ;NULL REMARK 3 REMARK 3 PARAMETER FILE 1 :protein_rep.param REMARK 3 PARAMETER FILE 2 :water_rep.param REMARK 3 TOPOLOGY FILE 1 :protein_no_cter.top REMARK 3 TOPOLOGY FILE 2 .top REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL SSBOND 1CYSL 23 CYSL 88 SSBOND 2 CYS L 134 CYS L 194 SSBOND 3CYSH 22 CYS H 96 SSBOND 4 CYS H 143 CYS H 199 CRYST1 51.627 132.089 77.247 90.00 98.88 90.00C121 8 ORIGX1 1.000000 0.000000 0.000000 0.00000 ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 0.000000 0.000000 1.000000 0 SCALE1 70 0.000000 0.003027 0.00000 SCALE2 0.000000 0.007571 0.000000 0.00000 SCALE3 0.000000 0.000000 0.013103 0.00000 Figure 1 provides the three-dimensional structure of the XAB1 Fab as obtained in Example 1. Figure 1A is a space-filling representation. Figure 1B is a cartoon representation. The heavy- and lightchain of the XAB1 Fab appears in dark and light grey, respectively.

Example 2. Crystal ure of the XAB1 FV complex with human IL-17A: analysis of the paratope for structure-guided affinity maturation (i) al and Methods Standard molecular biological protocols were used to obtain the XAB1 Fv antibody nt. In brief, the Fv was cloned and expressed in E. coli W3110 with a C-terminal hexahistidine tag on the heavy-chain and a C-terminal Strep-tag on the light-chain. The recombinant protein was purified by Ni-chelate chromatography.

W0 2014/122613 The XAB1 Fv nt complex with human lL-17A was then prepared using standard methodology. In brief, human lL-17A (1.1mg) was mixed with an excess of Fv (2.7mg) and the complex was run on a S100 size-exclusion chromatography, in 10mM TRIS pH 7.4, 25mM NaCl. The protein complex was then trated by ultra-filtration to 21 .2mg/ml and crystallized.

Standard llization protocols were followed. In brief, crystals were grown at 19°C in SD2 96 well-plates, using the method of vapour ion in sitting drops. The protein stock was mixed 1:1 with a crystallization buffer containing 10% PEG 20,000, 0.1M Bicine pH 9.0, 2.0% (v/v) dioxane. Total drop size was 0.4 ul. Prior to X-ray data 1O collection, one crystal was y transferred into a 1:1 mix of the crystallization buffer with 20% PEG 20,000, 30% glycerol, and then flash cooled into liquid nitrogen.

X-ray data collection and processing was carried out using standard protocols. Briefly, X-ray data to 3.0A resolution were collected at the Swiss Light Source, Beamline X1OSA, with a MAR225 CCD detector, using 1.0000A X-ray radiation. In total, 110 images of 10° oscillation each were recorded at a crystal-to-detector distance of 300mm and processed with the HKL2000 software package. The crystal belonged to space group P21212 with cell parameters a=184.31A, b=55.81A, c=70.99A, or = B = y = 90°. R-sym to 3.0A tion was 11.2% and data completeness 99.9%.

The structure was determined by molecular replacement with the program PHASER. A search model for the XAB1 Fv was generated from the crystal ure of the XAB1 Fab previously determined (see Example 1). A search model for lL-17A was generated from the published human lL-17F crystal structure (PDB entry 1jpy). lterative model building and refinement were performed with Coot allographic Object-Oriented Toolkit) and CNX (Crystallography & NMR eXplorer) version 2002, until no further significant improvements could be made to the model. Final R- and R-free for all data were 0.215 and 0.269, respectively. The final refined model showed a ean- square deviation (RMSD) from ideal bond lengths and bond angles of 0.007A and 1.00, respectively. (ii) Results The molecular replacement calculations revealed a dimeric complex comprising one |L- 17A homodimer with two XAB1 Fv fragments bound. The results of the X-ray refinement of the XAB1 Fv complex with human |L-17A are provided in Table 10 and the three- dimensional structure of this complex is shown in Figure 2. Each XAB1 Fv makes ts to both |L-17A ts, but the vast majority of the olecular ts (about 96% of the buried surface) are contributed by one |L-17A subunit only.

Table 10. X-ray refinement of the XAB1 Fv complex with |L-17A obtained by the program CNX.

REMARK 3 REMARK 3 REFINEMENT.

REMARK PROGRAM : CNX 2002 REMARK on(no:(no:wwwwe»we»wwwwwwwwwwwwwwwwwwwwwwwww AUTHORS : Brunger, Adams, Clore, Delano, REMARK Gros, Grosse-Kunstleve, Jiang, REMARK Kuszewski, Nilges, Pannu, Read, REMARK Rice, Simonson, Warren REMARK and REMARK ys |nc., REMARK (Badger, Berard, Kumar, Szalma, REMARK Yip, Dzakula).

REMARK REMARK DATA USED IN REFINEMENT.

REMARK RESOLUTION RANGE HIGH ROMS) : 3.01 REMARK RESOLUTION RANGE Low (ANGSTROMS): 47.74 REMARK DATA CUTOFF (SIGMA(F)) : 0.0 REMARK DATA CUTOFF HIGH (ABS(F)) : 1527617580 REMARK DATA CUTOFF Low (ABS(F)) : 0.000000 REMARK COMPLETENESS (WORKING+TEST) (%) : 99.5 REMARK NUMBER OF REFLECTIONS : 15190 REMARK REMARK FIT TO DATA USED IN REFINEMENT.

REMARK CROSS-VALIDATION METHOD : THROUGHOUT REMARK FREE R VALUE TEST SET SELECTION : RANDOM REMARK R VALUE (WORKING SET) : 0.215 REMARK FREE R VALUE : 0.269 REMARK FREE R VALUE TEST SET SIZE (%) : 4.9 REMARK FREE R VALUE TEST SET COUNT : 748 REMARK ESTIMATED ERROR OF FREE R VALUE : 0.010 REMARK REMARK FIT IN THE HIGHEST RESOLUTION BIN.

REMARK TOTAL NUMBER OF BINS USED : 6 REMARK BIN RESOLUTION RANGE HIGH (A) : 3.00 REMARK BIN TION RANGE LOW (A) : 3.19 REMARK BIN COMPLETENESS (WORKING+TEST) (%) : 94.6 REMARK REFLECTIONS IN BIN (WORKING SET) : 2234 REMARK BIN R VALUE (WORKING SET) : 0.301 REMARK BIN FREE R VALUE : 0.350 REMARK BIN FREE R VALUE TEST SET SIZE (%) : 5.3 REMARK BIN FREE R VALUE TEST SET COUNT : 124 REMARK TED ERROR OF BIN FREE R VALUE : 0.031 REMARK REMARK NUMBER OF NON-HYDROGEN ATOMS USED IN MENT.

REMARK PROTEIN ATOMS : 5007 REMARK NUCLEIC ACID ATOMS : 0 REMARK GEN ATOMS : 0 REMARK SOLVENT ATOMS : 33 REMARK REMARK B VALUES.

REMARK FROM WILSON PLOT (A**2) : 54.9 REMARK MEAN B VALUE (OVERALL, A**2) : 44.8 REMARK OVERALL ANISOTROPIC B VALUE.

REMARK B11 (A**2): 5.66 REMARK BZZ(A**2): 0.97 REMARK B33 (A**2) : -6.63 REMARK wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww B12(A**2): 0.00 REMARK B13(A**2): 0.00 REMARK BZ3(A**2): 0.00 REMARK REMARK BULK SOLVENT MODELING.

REMARK METHOD USED : FLAT MODEL REMARK KSOL : 0.313124 REMARK BSOL : 20.608 (A**2) REMARK REMARK ESTIMATED COORDINATE ERROR.

REMARK ESD FROM LUZZATI PLOT (A) : 0.33 REMARK ESD FROM SIGMAA (A) : 0.39 REMARK LOW RESOLUTION CUTOFF (A) : 5.00 REMARK REMARK CROSS-VALIDATED ESTIMATED COORDINATE ERROR.

REMARK ESD FROM C-V LUZZATI PLOT (A) : 0.44 REMARK ESD FROM C-V SIGMAA (A) : 0.51 REMARK REMARK RMS DEVIATIONS FROM IDEAL VALUES.

REMARK BOND LENGTHS (A) : 0.007 REMARK BOND ANGLES (DEGREES) : 1.0 REMARK DIHEDRAL ANGLES (DEGREES) : 22.1 REMARK IMPROPER ANGLES (DEGREES) : 0.78 REMARK REMARK ISOTROPIC THERMAL MODEL : RESTRAINED REMARK REMARK ISOTROPIC L FACTOR INTS. RMS SIGMA REMARK MAIN-CHAIN BOND (A**2) : 1.46 ; 1.50 REMARK MAIN-CHAIN ANGLE (A**2) : 2.62 ; 2.00 REMARK SIDE-CHAIN BOND (A**2) : 1.63 ; 2.00 REMARK SIDE-CHAIN ANGLE (A**2) : 2.62 ; 2.50 REMARK REMARK NCS MODEL : NONE REMARK REMARK NCS RESTRAINTS. RMS SIGMANVEIGHT REMARK GROUP 1 POSITIONAL (A): NULL ;NULL REMARK GROUP 1 B-FACTOR (A**2) : NULL ;NULL REMARK REMARK PARAMETER FILE 1 in_rep.param REMARK PARAMETER FILE 2 :water_rep.param REMARK TOPOLOGY FILE 1 :protein_no_cter.top REMARK TOPOLOGY FILE 2 :water.top REMARK REMARK OTHER REFINEMENT REMARKS: NULL W0 2014/122613 SSBOND 1 CYS L 23 CYS L 88 SSBOND 2 CYS H 22 CYS H 96 SSBOND 3 CYS A 23 CYS A 88 SSBOND 4 CYS B 22 CYS B 96 SSBOND 5 CYS C 94 CYS C 144 SSBOND 6 CYS C 99 CYS C 146 SSBOND 7 CYS D 94 CYS D 144 SSBOND 8 CYS D 99 CYS D 146 CRYST1 184.306 55.813 70.991 90.00 90.00 90.00 P 21 21 2 24 ORIGX1 1.000000 0.000000 0.000000 0.00000 ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 00 0.000000 1.000000 0.00000 SCALE1 0.005426 0.000000 0.000000 0.00000 SCALE2 0.000000 0.017917 0.000000 0.00000 SCALE3 0.000000 0.000000 0.014086 0.00000 Figure 2 provides the three-dimensional structure of the XAB1 Fv complex with human lL-17A, as obtained in e 2. Figure 2A shows the two XAB1 Fv fragments in space-filling representation; the lL-17A homodimer is shown in cartoon representation.

Figure 2B shows the two XAB1 Fv fragments in cartoon representation; the lL-17A homodimer is shown in space-filling representation. The heavy- and light-chain of the XAB1 Fv are represented as dark and light grey, respectively. One chain of the lL-17A homodimer is represented as light grey, the other is represented as dark grey.

A ed analysis of the complex was performed. A careful visual inspection of the crystal structure with the programs Coot and Pymol was carried out, and the amount of protein surface buried at the antibody-antigen interface was calculated with the program AREAIMOL of the CCP4 m suite. Intermolecular contacts were defined using a cut-off distance of 3.9A between antibody and n atoms. An overview of binding can be summarized as follows. Binding of XAB1 is symmetrical; each Fv nt binds to an equivalent epitope on the |L17A homodimer.

The binding of each Fv nt buried on average 1732A2 of ed surface, and involved 30 antibody and 25 lL-17A amino acid residues. The contribution to the buried surface of the XAB1 chain (around 56OA2) was r than that of the heavy chain (around 275A2). In on, CDRH2 did not make any direct contacts to lL-17A and appeared to be too far from the protein antigen to provide opportunities for affinity maturation. The CDRH1 contribution appeared to be limited to one amino-acid side- chain only (Tyr32); this CDR was also too far from lL-17A to offer opportunities for affinity maturation through amino acid substitutions. The XAB1 CDRH3 made multiple tight contacts with lL-17A. r, careful inspection of the structure in this region W0 2014/122613 failed to reveal any opportunity for further enhancing these contacts by point mutations; therefore, CDRH3 was deemed unsuitable as a target region for affinity enhancement.

In contrast, tion of the light-chain CDRs showed multiple opportunities for affinity maturation. Among the three light-chain CDRs, CDRL1 was considered most promising, and based on this observation, the inventors proposed to randomize positions 30 to 32 of the light-chain in an attempt to strengthen contacts to lL-17A residues Arg124, Phe133 and Tyr85.

Affinity maturation by rational design 1O Based on the results above, it was seen that the XA81 interface with homodimeric lL- 17A was comparatively small and was characterized by a dominant contribution from the light chain, no ement from CDRH2, and mainly indirect contribution by CDRH1 (i.e. via stabilization of . ingly, the heavy chain of XA81 did not appear to offer promising opportunities for ty maturation.

In st, the XA81 light chain did offer some opportunities in amino acid residues 30 to 32, with an optional insertion of up to 4 amino acid residues (CDRL1), amino acids 51 to 53 and 56 (CDRL2) and amino acid residues 92 and 93, with an optional insertion of up to 4 amino acid residues.

The availability of the published crystal ure for homodimeric human lL-17F, and 2O the structure of homodimeric lL-17F in complex with the human receptor lL-17RA allowed for predictions to be made based on the ed structures of crystallized lL- 17A and lL-17A in complex with XA81 (and variants thereof).

The structural similarities predicted between lL-17F and lL-17A (on the basis of ce identity and homology) were investigated. lL-17F and lL-17A bore a structural resemblance. The inventors hypothesized that lL-17A would bind to the N-terminal domain of its receptor in the same manner as has been shown for the published lL- 17F/lL-17RA complex (Ely LK et al 2009, Nat l. 10:1245-51).

W0 2014/122613 On the basis of the observed structure and comparison of the known sequences for human lL-17A and lL-17F, along with the sequence of lL-17A derived from other species, a number of additional predictions were made by the inventors: It was ed that XA81 (and antibodies variants derived there from having an improved affinity for the epitope targeted by XABi) would be highly specific for human lL-17A. It was hypothesized that such antibodies would retain some reactivity with lL-17A from other s (on the basis of the high degree of conserved sequence identity or homology between s). However, on the basis of the available sequence data and structural predictions it was not clear to what extent reactivity 1O with species variants of lL-17A could be expected. Given the lack of structural similarity with other lnterleukins, cross-reactivity with such molecules (from humans or other s) was expected to be very unlikely.

In addition, differences between the sequences of lL-17A and lL-17F (in particular N- terminal region) gave rise to predictions that the L-17A antibodies of the disclosure would not bind to lL-17F. For example, overlays of the two crystal structures indicated that steric hindrance would prevent binding between these antibodies and lL-17F.

Furthermore, an extrapolation to the structure of lL-17AF heterodimers also suggested that such interference, in particular in the inal , would hamper binding of the antibodies to lL-17AF heterodimers and thereby result in a lack of binding to lL- 17AF, Le. a lack of cross-reactivity by these antibodies for lL-17AF dimers.

Example 3. Generation of ty d antibody variants Actual affinity maturation of the initial antibody XA81 focused on the light chain, for reasons discussed above. The work was carried out in three steps: (i) library generation, (ii) library screening, and (iii) candidate characterisation.

The protein engineering work (i.e. affinity maturation) was carried out in the Fab fragment format for ease of handling. Candidates were formatted back to full lgG after engineering.

W0 2014/122613 (i) Library generation The DNA sequence ng the variable domain of the light chain was mutated to create a library of gene variants. Two different approaches (A and B) were used for library generation, providing two separate libraries. 1) Method A — random mutation by error prone PCR: The DNA region ng the variable domain of the light chain of XABl was randomly mutated using error prone PCR. In more detail, this region was amplified using the polymerase Mutazyme II, which uced mutations at a high ncy (for more detail, see the guidance supplied with the GeneMorph ll random nesis kit, 1O supplied by Stratagene #200550). However, any suitable random mutation technique or strategy could be used.

The pool of PCR fragment variants was then cloned by cutting and g into the expression vector of XABl. Essentially, the parent, unmutated sequence was cut out of the expression vector and replaced by a randomly mutagenized sequence which was pasted in its place. Standard molecular biology techniques were used to accomplish this.

This resulted in a library of expression vector variants comprising a variety of randomly mutagenized variable domain ces. 2) Method B — mutation by rational design: Under this approach, the generation of the library was guided by the structural analysis carried out as a precursor to affinity maturation. Specific amino acid es (in particular in CDR1 of the light chain of XABl) were targeted based on the e and paratope information derived from the crystal structure described above.

Three amino acid residues, ed on the basis of the crystal structure information, were fully randomised. Standard molecular biology approaches were used for the construction.

Firstly, a fragment of the variable region encoding the riate CDR and a first part of the light chain framework was amplified by PCR, using degenerate oligonucleotides.

W0 22613 That is, the oligonucleotides, ng the CDR were synthesised in such a way as to provide a variety of bases at a defined position or positions. Design of the oligonucleotide enabled randomization of specifically targeted amino acid positions in the CDR by NNK degenerated codons (in which N stands for all 4 bases, A, T, C and G and K for G and T) and allowed all 20 natural amino acids at those positions.

Following this first step, a second fragment overlapping the first one and encoding the remaining part of the light chain, was also amplified by PCR. Both fragments were then led by an "assembly" PCR to generate the complete le light chain and cloned back into the expression vector in a ‘cut and paste’ manner. Thereby the 1O parental sequence was replaced with a range of rationally mutated sequences, whereby at specific amino acid positions all 20 natural amino acids were represented. (ii) Library screening Once libraries comprising sequences encoding XA81 variants had been ted it was necessary to screen them in order to select those which had superior characteristics to the parental XA81 sequence, for example higher affinity for lL-17A.

Two screening techniques were used. Firstly, a high throughput screening was done by "colony filtration screening" (CFS). This assay permitted a ient screening of large number of clones. It allowed reduction to positive hits prior to ELISA screening, which was useful in particular for the random approach "method A" as the y size was much larger (>105) compared to the library size in d B" (only 8000). ELISA screening is convenient for 104 clones or less and gives more quantitative results. 1) Colony filtration screening (CFS): The ol for CFS was based on Skerra et al. 1991, Anal Biochem 196:151—155.

Some adaptations were made.

E. coli colonies expressing the Fab variant libraries were grown on a filter on top of a Petri dish containing LB agar and glucose. In parallel, a PVDF membrane was coated with the target protein (IL-17A). The coated membrane was placed on the agar plate.

The filter with colonies of Fab fragment expressing E. coli was placed on top of the ne. The Fab fragments expressed by the cells diffused from the colonies and W0 2014/122613 bound the target lL-17A. The Fab fragment thus captured on the PVDF membrane was then detected using a secondary antibody ated with alkaline phosphatase for Western staining. The ions for selecting only variants with improved binding properties were previously established using XAB1 as reference.

More ically, after transformation of E. coli cells with the library, the cells were spread on a DuraporeTM membrane filter (0.22pm GV, Millipore®, cat #GVWP09050) placed on a Petri dish containing LB agar + 1% glucose + antibiotic of interest. The plates were incubated overnight at 30°C The PVDF membrane (lmmobilon-P, Millipore®, cat #IPVH08100) was pre-wet in 1O methanol, washed in PBS and coated with a hulL-17A solution at 1 ug/ml in PBS. The membrane was incubated overnight at room temperature. After coating, the membrane was washed 2 times in Tris buffered saline (TBS) + 0.05% Tween (TBST) and blocked two hours at room temperature in 5% milk TBST. Then, the membrane was washed four times in TBST and soaked in 2xYT medium with 1mM IPTG. This membrane, called the capture membrane was placed onto a LB agar plate with 1mM IPTG + antibiotic of interest, and was covered with the Durapore membrane with the colonies on top. The ing ch was incubated four hours at 30°C.

After this incubation, the capture membrane was washed 4 times with TBST and blocked in 5% milk TBST for 1 hour at room ature. Then, the membrane was washed once with TBST and incubated with a secondary antibody (anti-hu_kappa light chain antibody, alkaline-phosphatase (AP) conjugated, Sigma # A3813, diluted 1:5000 in 2% milk TBST), 1 hour at room temperature. Afterward, the membrane was washed 4 times in TBST, once in TBS and incubated in the substrate on (SigmaFast BT , 1 tablet in 10ml H20). When the signal reached the expected ity the membrane was washed with water and allowed to dry.

After development of the signal on the capture membrane, the colonies giving stronger signal than the parental XAB1 were picked and allowed to proceed to a secondary ELISA screening described below.

W0 22613 2) ELISA Screening: Following the CFS, ELISA was used to screen the candidates selected by CFS. In brief, for the relative low number of variants identified by error-prone PCR mutagenesis (i.e. library A) the ELISA was performed manually in a 96 well format. In contrast, for the ies constructed by rational design (method B), a larger number of ed clones needed to be screened at ELISA level to be able to discriminate between their different binding affinity to IL-17A and identify the clones with the highest affinity. An ELISA robot was used for that purpose in a 384 well plate format. However, the ELISA protocol was the same in each case, the only difference being the volumes of reagents. 1O a) Cell cultures: Clones were first grown overnight at 30°C, 900rpm, in 2xYT medium + 1%glucose + antibiotic of interest. The plates containing these cultures were called master plates.

The next day, ts of cultures from the master plates were transferred into expression plates containing 2xYT medium + 0.1% glucose + antibiotic of st.

These plates were incubated at 30°C, 900rpm about 3 hours. Then isopropyl B-D thiogalactopyranoside (IPTG) on was added to a final concentration of 0.5mM. The plates were incubated overnight at 30°C, 990 rpm.

The next day, lysis buffer (2x) Borate buffered saline (BBS) solution (Teknova #80205) + 2.5mg/ml lysosyme + 10u/ml Benzonase) was added to the cultures. Plates were incubated 1 hour at room temperature, then 12.5 % milk TBST was added for blocking.

After 30 min incubation, cells lysates were diluted 1:10 in 2% milk TBST and were transferred into the ELISA plates. b) ELISA: ELISA plates (Nunc Maxisorp) were coated with a hulL-17A solution at 1 ug/ml during 1 hour. The plates were washed once with TBST and blocked 1 hour with 5% milk TBST.

After ng, plates were washed 3 times with TBST and then, d cell lysates were loaded on the plates and incubated 1 hour. Afterward, plates were washed 3 times with TBST and were incubated 1 hour with a secondary antibody AP conjugated.

W0 2014/122613 1 00 The plates were finally washed 3 times with TBST and then incubated with the substrate solution (AttoPhos ate Set, Roche #11 681 982 001). The whole process was performed at room ature.

In addition to the "classic" ELISA bed above, modified ELISA were also undertaken for a better discrimination between clones with very high affinity (in the pico- molar range) for the target protein. An "off-rate" ELISA and a "competition" ELISA were developed for this purpose, as detailed below. c) "Off-rate" ELISA: For this assay, the modification compared to the "classic" ELISA protocol regarded the washing step after the binding step (incubation of cell lysate in ELISA plates). In the "classic" protocol, the plate was washed 3 times with TBST. The washing solution was dispensed and immediately aspirated, without any tion time. For the "off-rate" ELISA, the plate was washed 6 times during at least 3 hours. This long wash increased the stringency of the assay, and allowed fying clones with a slow off-rate. d) "Competition" ELISA: This modified ELISA protocol included an extra step after the binding step. After tion of cell , the plates were washed 3 times with TBST and then, a solution of the parental XA81 (200 nM in 2% milk TBST) was incubated overnight at room temperature. This long incubation with an excess of the parental Fab allowed, as in the case of "off-rate" ELISA, to identify clones with slow off-rate, which lead to better discrimination between clones with an affinity in the picomolar range. The rest of the protocol was similar to the "classic" ELISA protocol. The secondary antibody used in this case was an AP conjugated anti-Flag tag antibody, since the Fabs ts from the library had a Flag tag at the C-terminus of the heavy chain but not the parental XA81 Fab used for the competition. (iii) Candidate characterisation The hits identified during the screening were produced on a larger scale for further physicochemical characterisation and to m high affinity binding to IL-17A, and/or W0 2014/122613 1 01 other advantageous properties in additional assays. These are described below in more detail. (iv) Results: ing and l characterization of candidates fol/owing ah‘inity maturation ofXAB1 1) Random mutagenesis approach (method A): The mutation rate after the error-prone PCR library generation was found to peak at 2 to 3 mutations per gene. Around 3x104 clones were screened by colony filter screening and a number of 94 clones were identified as improved and allowed to proceed to 1O binding, off-rate and competition ELISA. ELISA data in combination with cing results led to the identification of 6 candidates highlighting 3 potential hot spots for improvement, Gly at position 28 to Val (G28V) in LCDR1; Gly at on 66 to Val (G66V) or Ser (G668) in framework 3; Asn92 to Asp (N92D) in LCDR3 (data not shown, but oning is identical to that of XAB2, VL, i.e. SEQ ID NO: 25).

A stop codon was observed in one of the clones, but was not relevant as the E. coli strain used was an amber suppressor strain allowing read-through. Based on the data obtained, a G28V and G66V mutation appeared to cause the best ement. A variant of XAB1 was generated by standard molecular biology techniques carrying the two point mutations mentioned. A further variant was cloned having the N92D 2O substitution in addition, in order to test whether the l of the potential post- translational deamidation site (N92, S93) would be beneficial. More detailed profiling was done on those two variants, in particular of the triple mutant variant referred to as XAB_A2 which finally led to XAB2. ln XAB2, amino acids number 1 to 23 according to the Kabat tion constitute framework 1, amino acids number 24 to 34 (Kabat) constitute LCDR1, amino acids number 35 to 49 (Kabat) constitute framework 2, amino acids 50 to 56 (Kabat) constitute LCDR2, amino acids 57 to 88 (Kabat) constitute framework 3, amino acids 89 to 97 (Kabat) constitute LCDR3 and amino acids 98 to 107 (Kabat) tute framework 4. The same ision of other VL sequences according to embodiments of the disclosure also applies.

W0 2014/122613 1 02 Thus, the G66V substitution mentioned above is in a framework region, which is called the outer loop. This framework region is able to contribute to binding in some cases.

Based on the available structural information it was retrospectively suggested that this on indeed might be able to interact with a region of IL-17A which cannot be resolved from the crystal structure but may be in proximity to the outer loop. 2) Rational mutagenesis ch (method B): A snapshot of the amino acid distribution at the randomized positions was generated by sequencing of 32 randomly picked members. There was no significant bias, though statistics with this low number of sequences cannot be done. Around 4x104 clones were screened which oversampled the theoretical library size of 8000. A high number of hits were identified and 2630 clones were allowed to d to ELISA screening. ming binding, off-rate and competition ELISA, 60 clones with the highest ements were ced. In those 60 clones 22 unique sequences were found, and the result is summarized in Table 11.

Table 11. ELISA of all selected 22 unique candidates. Values are normalized to parental Fab XAB1. The representation indicates how often a certain sequence was found within the 60 hits. The difference in amino acid sequence is given in the three last s. XAB1 has the amino acids | S A at those positions. ELISA signals determined from crude extract of Fab expression culture from E. coli.

Candidate Classic Off- Compe- Represent- name ELISA rate titon ELISA ation % ELISA W0 2014/122613 1 O3 —3:33: --—— —--——--n Of the 22 unique clones, 6 were selected for 0.5 L scale standard E. coli expression and two step purification by IMAC (Ni-NTA) and SEC. Purified Fabs were then used to confirm the ement in binding by ELISA.

ELISA results of selected and ed Fab candidates in comparison to XA81 are shown in Figure 3, where the graph numbering corresponds to the candidate designation as follows: 1 is M8440; 2 is M8464; 3 is M8468; 4 is M8444; 5 is M8435; 6 is M8463; 7 is XA81.

Figure 3A is a graph showing the normalized signal versus the Fab concentration (M). It 1O can be seen that all the selected clones resulted in a higher signal than XA81. Figure 38 is a graph g the normalized remaining signal versus the washing incubation time ). All the selected clones result in a higher signal than XA81. Figure 3C is a graph showing the normalized signal versus the Fab competitor concentration (M).

Again, it can be seen that all the selected clones result in a higher signal than XA81.

W0 2014/122613 1 04 Example 4. Targeting a potential post-translational deamidation site.

The inventors hypothesized that the amino acid motif gine ed by glycine (NG) or, to lower extend also when followed by serine (NS), may be susceptible to post- translational deamidation. Such motifs are present in L-CDR2 (position 56/57) and L- CDR3 (92/93) of the antibody XAB1. Four lgG variants were generated in order to test whether the NG site could be removed without affecting binding and activity ties.

These four point mutation variants were cloned by standard molecular biology procedures and produced by standard transient transfection of HEK cells in 100 ml scale and purified via a protein A column. 1O Purified lgG variants were analyzed in an in vitro neutralization assay (e.g. as described in examples 12 and 13) to compare their activity to the parental XAB1 lgG. Results showed that out of these four ts, three had a reduced activity. But the candidate XAB_B12 (mutation N56Q) retained activity compared to the parental XAB1.

Table 12. ew of sequence modifications to XAB1, and corresponding effect on in vitro neutralization.

Kabat CDR L-CDR2 -__050(nM) e# 49 5O 51 52 53 54 55 56 57 lL-6 lL-8 Kabat# 49 5O 51 52 53 54 55 56 57 lgG Generic variants name XAB_G57T XAB_N56Q XAB_N56T XAB_N56S W0 2014/122613 1 05 Having thus identified the most suitable substitution, it was introduced to the most ing hits identified during the ty maturation process, resulting in XABZ (XAB_A2 N560), XABB (MB468 N560), XAB4 (MB435 N560). They were produced by standard transient transfection of HEK cells and purified via protein A column along with XAB5 (MB435), which still carried the NG site.

The NG motif was removed (N560) for the XABZ, XABS, XAB4, but was still present in XAB5. The NS motif in L-CDR3 was removed (N92D) in XABZ, as found during the random affinity maturation approach. Therefore, an optimal set of variants was available to test the susceptibility for deamidation of the potential sites. 1O The four purified candidates were d in a buffer pH 8 and incubated at 40°C in order to force the deamidation reaction. Aliquots were taken at several time points to ine the degree of deamidation by cation ge chromatography (CEX), according to principles well known to a person d in the art, and the in vitro neutralization activity by a cell based assay was determined (e.g. as described in es 12 and 13).

CEX results showed an increase of acidic variants percentage over time, as expected for any lgG, likely due to post-translational cation sites in the antibody framework, but the extent of increase was higher for XAB5 than for the other candidates, Le. 72% vs. 46% after one week and 94% vs. 83% after 4 weeks. Finally, in vitro neutralization activity assay results correlated with the CEX results, showing that XAB5 lost activity after 4 weeks incubation during forced deamidation condition. Size-exclusion chromatography-multi angle light scattering methodology (SEC-MALS), well known to a person skilled in the art, was used to monitor the aggregation levels in the samples.

W0 2014/122613 1 06 The data is summarized in Table 13.

Table 13. Analysis by SEC-MALS, in Vitro neutralization activity and CEX.

Anti M'6" E050 b) CEXC) [%] M '60 [%] E050 b) CEXC) [%] body [%] [rig/ml] [rig/ml] T: 0 weeks T: 1 weeks XABZ 99 45 15 98 no. 45 XA83 99 4o 14 98 no. 44 XAB5 99 45 18 98 no. 72 XAB4 99 48 15 98 no. 48 Anti M'60 EC50 b) CEXC) [%] NGd) NSd) body [%] [ng/ml] sites sites T: 4 weeks XABZ 95 47 85 o o XA83 97 4o 81 o 1 XAB5 94 61 94 1 1 XAB4 94 47 84 o 1 "monomer by SEC-MALS, b) inhibition of lL-6 secretion after cell stimulation with 80 ng/ml lL-17, "acidic ts by exchange chromatography, d’number of sites in CDRs (framework region not considered) These data indicated successful removal of a potential post-translational deamidation site, which could have had an effect on antibody ty. This is advantageous, since XABZ, XABB and XAB4 are therefore likely to achieve a more neous t than XA81 as no post-translational deamidation can occur during production or storage affecting the antibody activity.

W0 2014/122613 1 07 Example 5. X-ray analysis of antibody variants derived by affinity maturation: XAB2 In brief, the XAB2 Fv was cloned and expressed in E. coli TGf1- with a C-terminal stidine tag on the heavy-chain and a C-terminal Strep-tag on the light-chain, according to principles well known to a person skilled in the art. The recombinant protein was purified by Ni—chelate chromatography and size-exclusion chromatography (SPX-75).

The XAB2 Fv fragment complex with human lL-17A was then prepared using standard methodology. In brief, human lL-17A ) was mixed with an excess of XAB2 Fv 1O (3.7mg) and the complex was run on a S100 size-exclusion chromatography, in 10mM TRIS pH 7.4, 25mM NaCl. The protein complex was then concentrated by ultra-filtration to 26.3mg/ml and crystallized.

Standard crystallization protocols were followed. In brief, crystals were grown at 19°C in SD2 l plates, using the method of vapour diffusion in sitting drops. The protein stock was mixed 1:1 with a crystallization buffer containing 0.2M calcium acetate, 20% PEG 3,350. Total drop size was 0.4 ul. Prior to X-ray data collection, one crystal was briefly transferred into a 1:1 mix of the crystallization buffer with 30% PEG 3,350, 30% glycerol, and then flash cooled into liquid nitrogen.

X-ray data tion and sing was carried out using standard protocols. Briefly, X-ray data to 2.0A resolution were collected at the Swiss Light Source, beamline X06DA, with a MAR225 CCD or, using 1.0000A X-ray radiation. In total, 360 images of 05° oscillation each were recorded at a crystal-to-detector distance of 190mm and processed with the XDS software package. The crystal belonged to space group P21212 with cell parameters a=184.72A, 6A, c=71.11A, OL = B = y = 90°. R- sym to 2.0A resolution was 5.2% and data completeness 100.0%.

As the l of the XAB2 Fv complex was highly isomorphous with the crystal of the XAB1 Fv complex (Example 2), the structure of the latter was used as input model for an initial run of crystallographic ment with the program CNX. Iterative model correction and refinement was med with Coot (Crystallographic -Oriented Toolkit) and CNX (Crystallography & NMR eXplorer) version 2002, until no further W0 2014/122613 1 08 significant improvements could be made to the crystallographic model. Final R- and R- free for all data were 0.214 and 0.259, tively. The final d model showed a root-mean-square deviation (RMSD) from ideal bond lengths and bond angles of 0.005A and 09°, respectively.

Results The results of the X-ray refinement of the XAB2 Fv complex with human |L-17A are provided in Table 14 and the three-dimensional structure of this complex is shown in Figure 4. The X-ray crystallography analysis confirmed that the variant dy XAB2 retained the target specificity and bound with high affinity to essentially the same 1O epitope as the parental XAB1 antibody. However, in the XAB1 complex structure, the light-chain loop comprising Gly66 adopts a conformation that is no longer possible when this residue is mutated to a valine. As a consequence, in the XAB2 complex, the Gly66 to valine mutation (G66V) forces the loop to adopt a new conformation, and the valine side-chain makes hydrophobic contacts to lle51 of |L-17A (Figure 5). Two more |L-17A residues, Pro42 and Arg43, become visible (ordered) in this crystal structure. These antigen residues make additional binding interactions with the XAB2 antibody, in particular hobic contacts to Val28 (Figure 5).

Table 14. X-ray ment of the XABZ Fv x with |L-17A obtained by the program CNX.

REMARK 3 REMARK EMENT.

REMARK 3 PROGRAM : CNX 2002 REMARK 3 AUTHORS :Brunger, Adams, Clore, Delano, REMARK 3 Gros, Grosse-Kunstleve, Jiang, REMARK 3 ski, Nilges, Pannu, Read, REMARK 3 Rice, Simonson, Warren REMARK 3 and REMARK 3 Accelrys Inc., REMARK 3 (Badger, Berard, Kumar, Szalma, REMARK 3 Yip, a).

REMARK 3 REMARK 3 DATA USED IN REFINEMENT.

REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.00 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS): 71.11 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.0 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 232935020 REMARK 3 DATA CUTOFF LOW (ABS(F)): 0.000000 REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 99.8 WO 22613 REMARK NUMBER OF REFLECTIONS : 50409 REMARK REMARK FIT TO DATA USED IN REFINEMENT.

REMARK CROSS-VALIDATION METHOD : HOUT REMARK FREE R VALUE TEST SET SELECTION : RANDOM REMARK R VALUE (WORKING SET) : 0.214 REMARK FREE R VALUE : 0.259 REMARK FREE R VALUE TEST SET SIZE (%) : 5.0 REMARK FREE R VALUE TEST SET COUNT : 2521 REMARK ESTIMATED ERROR OF FREE R VALUE : 0.005 REMARK REMARK FIT IN THE HIGHEST RESOLUTION BIN.

REMARK TOTAL NUMBER OF BINS USED : 6 REMARK BIN TION RANGE HIGH (A) : 2.00 REMARK wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww BIN RESOLUTION RANGE LOW (A) : 2.13 REMARK BIN COMPLETENESS (WORKING+TEST) (%) :100.0 REMARK REFLECTIONS IN BIN (WORKING SET) : 7858 REMARK BIN R VALUE (WORKING SET) : 0.262 REMARK BIN FREE R VALUE : 0.304 REMARK BIN FREE R VALUE TEST SET SIZE (%) : 5.0 REMARK BIN FREE R VALUE TEST SET COUNT : 414 REMARK ESTIMATED ERROR OF BIN FREE R VALUE : 0.015 REMARK REMARK NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.

REMARK PROTEIN ATOMS : 5055 REMARK NUCLEIC ACID ATOMS : 0 REMARK HETEROGEN ATOMS : 0 REMARK SOLVENT ATOMS : 376 REMARK REMARK B VALUES.

REMARK FROM WILSON PLOT (A**2) : 27.8 REMARK MEAN B VALUE (OVERALL, A**2) : 37.3 REMARK OVERALL ANISOTROPIC B VALUE.

REMARK B11 (A**2) : -0.85 REMARK BZZ(A**2): 3.93 REMARK B33 (A**2) : -3.08 REMARK B12(A**2): 0.00 REMARK B13(A**2): 0.00 REMARK *2): 0.00 REMARK REMARK BULK SOLVENT MODELING.

REMARK METHOD USED : FLAT MODEL REMARK KSOL : 94 REMARK BSOL : 46.0594 (A**2) REMARK REMARK ESTIMATED COORDINATE ERROR.

REMARK ESD FROM LUZZATI PLOT (A) : 0.25 REMARK ESD FROM SIGMAA (A) : 0.19 REMARK LOW RESOLUTION CUTOFF (A) : 5.00 REMARK REMARK CROSS-VALIDATED ESTIMATED COORDINATE ERROR.

REMARK ESD FROM C-V LUZZATI PLOT (A) : 0.31 REMARK ESD FROM C-V SIGMAA (A) : 0.22 REMARK REMARK RMS DEVIATIONS FROM IDEAL VALUES.

REMARK BOND LENGTHS (A) : 0.005 REMARK BOND ANGLES (DEGREES) : 0.9 REMARK DIHEDRAL ANGLES (DEGREES) : 21.0 REMARK IMPROPER ANGLES (DEGREES) : 0.70 REMARK REMARK 3 ISOTROPIC THERMAL MODEL: RESTRAINED REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR INTS. RMS SIGMA REMARK 3 MAIN-CHAIN BOND (A**2): 1.49; 1.50 REMARK 3 MAIN-CHAIN ANGLE (A**2): 2.44; 2.00 REMARK 3 SIDE-CHAIN BOND (A**2): 1.95; 2.00 REMARK 3 SIDE-CHAIN ANGLE (A**2): 2.93; 2.50 REMARK 3 REMARK 3 NCS MODEL : NONE REMARK 3 REMARK 3 NCS RESTRAINTS. RMS SIGMANVEIGHT REMARK 3 GROUP 1 POSITIONAL (A): NULL ;NULL REMARK 3 GROUP 1 OR (A**2) : NULL ;NULL REMARK 3 REMARK 3 PARAMETER FILE 1 :protein_rep.param REMARK 3 PARAMETER FILE 2 :water_rep.param REMARK 3 PARAMETER FILE 3 :ion.param REMARK 3 TOPOLOGY FILE 1 :protein.top REMARK 3 TOPOLOGY FILE 2 :water.top REMARK 3 TOPOLOGY FILE 4 op REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL SSBOND 1 CYS L 23 CYS L 88 SSBOND 2 CYS H 22 CYS H 96 SSBOND 3 CYS A 23 CYS A 88 SSBOND 4 CYS B 22 CYS B 96 SSBOND 5 CYS C 94 CYS C 144 SSBOND 6 CYS C 99 CYS C 146 SSBOND 7 CYS D 94 CYS D 144 SSBOND 8 CYS D 99 CYS D 146 CRYST1 184.719 55.558 71.109 90.00 90.00 90.00 P 21 21 2 24 ORIGX1 1.000000 0.000000 0.000000 0.00000 ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 00 0.000000 1.000000 0.00000 SCALE1 0.005414 0.000000 0.000000 0 SCALE2 0.000000 0.017999 0.000000 0.00000 SCALE3 0.000000 0.000000 0.014063 0.00000 Figure 4 provides the three-dimensional structure of the XAB2 Fv complex with human lL-17A. Figure 4A shows the two XAB2 Fv fragments in filling representation, and the lL-17A homodimer is shown in cartoon representation. Figure 4B shows the two XAB2 Fv fragments in cartoon representation, and the lL-17A homodimer is shown in space-filling representation. The heavy- and light-chain of the XAB2 Fv are shown in dark and light grey, respectively. One chain of the lL-17A mer is shown in light grey, the other is shown in dark grey.

Figure 5 provides the three-dimensional ure of the XAB2 Fv complex with human lL-17A as a close-up view of the antibody L-CDR1 and outer loop regions, bearing the glycine to valine ons (G28V and G66V, respectively). The G66V mutation leads to W0 2014/122613 1 1 1 a change in the conformation of the outer loop, as well as to additional antibody-antigen contacts to lL-17A residues Pro42, Arg43 and lle51. The XAB2 Fv is represented in light-grey cartoon, and the human lL-17A homodimer in darker shades of grey. lle51 does not belong to the same lL-17A subunit as Pro42 and Arg43.

Example 6. X-ray analysis of antibody variants d by affinity maturation: XAB5 The XAB5 Fv was cloned and expressed in E. coli TGf1- with a C-terminal hexahistidine tag on the heavy-chain and a C-terminal Strep-tag on the light-chain. The recombinant protein was purified by Ni-chelate chromatography followed by size-exclusion 1O chromatography on a SPX-75 column, in PBS buffer. LC-MS analysis showed the expected mass for the heavy-chain (13703.4Da), and the presence of two forms of the light-chain: full-length (115aa; 12627.3Da; ca. 27%) and with truncated Strep-tag (A1 to 0112; 12222.8Da; ca. 73%).

The XAB5 Fv fragment complex with human lL-17A was then prepared using standard methodology. In brief, human lL-17A (1.4mg) was mixed with an excess of XAB5 Fv ) and the x was run on a S100 size-exclusion chromatography, in 10mM TRIS pH 7.4, 25mM NaCl. The protein complex was then concentrated by ultra-filtration to 16.5mg/ml and llized.

Standard crystallization protocols were followed. In brief, crystals were grown at 19°C in SD2 96 well-plates, using the method of vapour diffusion in sitting drops. The n stock was mixed 1:1 with a crystallization buffer containing 15% PEG 5,000 MME, 0.1M MES pH 6.5, 0.2M ammonium sulfate. Total drop size was 0.4 ul. Prior to X-ray data collection, one l was y transferred into a 1:1 mix of the crystallization buffer with 20% PEG 5,000 MME, 40% glycerol, and then flash cooled into liquid nitrogen.

X-ray data tion and processing was d out using standard protocols. Briefly, X-ray data to 3.1A resolution were collected at the Swiss Light Source, beamline X1OSA, with a Pilatus or, using 1.00000A X-ray radiation. In total, 720 images of 025° oscillation each were recorded at a crystal-to-detector distance of 520mm and processed with the XDS software package. The crystal belonged to space group C2221 W0 2014/122613 1 12 with cell parameters a=55.37A, b=84.08A, c=156.35A, or = B = y = 90°. R-sym to 3.1A resolution was 8.9% and data completeness 99.7%.

The structure was ined by molecular replacement with the program , using search models derived from the previously-determined XAB2 Fv complex. ive model correction and refinement was performed with Coot (Crystallographic Object-Oriented Toolkit) and CNX (Crystallography & NMR eXplorer) version 2002, until no further significant improvements could be made to the crystallographic model Final R- and R-free for all data were 0.222 and 0.305, respectively. The final d model showed a root-mean-square deviation (RMSD) from ideal bond lengths and bond 1O angles of 0.008A and 1.20, respectively.

Results The results of the X-ray refinement of the XAB5 Fv complex with human lL-17A are provided in Table 15 and the three-dimensional ure of this complex is shown in Figure 6. In this crystal structure, the XAB5 Fv complex has exact crystallographic 2-fold symmetry: the asymmetric unit of the crystal contains only one half of the whole, dimeric complex. The XAB5 Fv makes contacts to both lL-17A ts, but the vast majority of the intermolecular contacts are to only one subunit (around 90% of the lL-17A surface buried by the XAB5 Fv is contributed by one lL-17A subunit). The X-ray crystallography analysis confirmed that the variant antibody XAB5 retained the target specificity and bound with high affinity to essentially the same epitope as the parental XAB1 antibody.

However, in the XAB5 complex ure, the light-chain CDRL1 bears three point mutations which provide enhanced binding to human lL-17A. Trp 31 of the XAB5 light- chain is engaged in strong hydrophobic/aromatic interactions with Tyr 85 of lL-17A and, to a lesser extent, Phe 133 of lL-17A. Asn 30 of the XAB5 light-chain donates a H-bond to the main-chain carbonyl of Pro 130 of lL-17A and is in van der Waals contact to Leu 49 (same lL-17A t) and Val 45 (other lL-17A subunit). Glu 32 of the XAB5 light- chain stabilizes the CDRL1 loop through intramolecular ed interactions.

Furthermore, Glu 32 makes favorable electrostatic interactions with Arg 124 of lL-17A, but is not engaged into a "head-to-head" salt-bridge ction (Figure 7).

W0 2014/122613 1 13 Table 15. X-ray refinement of the XAB5 Fv complex with lL-17A obtained by the program CNX.

REMARK 3 REMARK EMENT.

REMARK 3 M : CNX 2002 REMARK 3 AUTHORS : Brunger, Adams, CIore, Delano, REMARK 3 Gros, -Kunstleve, Jiang, REMARK 3 Kuszewski, Nilges, Pannu, Read, REMARK 3 Rice, Simonson, Warren REMARK 3 and REMARK 3 Accelrys Inc., REMARK 3 (Badger, Berard, Kumar, Szalma, REMARK 3 Yip, Dzakula).

REMARK 3 REMARK 3 DATA USED IN REFINEMENT.

REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 3.11 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS): 46.25 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.0 REMARK 3 DATA CUTOFF HIGH (ABS(F)): 3778977.84 REMARK 3 DATA CUTOFF LOW (ABS(F)) : 0.000000 REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 99.0 REMARK 3 NUMBER OF REFLECTIONS : 6801 REMARK 3 REMARK 3 FIT TO DATA USED IN MENT.

REMARK 3 VALIDATION METHOD : THROUGHOUT REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM REMARK 3 R VALUE (WORKING SET) : 0.222 REMARK 3 FREE R VALUE : 0.305 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.0 REMARK 3 FREE R VALUE TEST SET COUNT : 340 REMARK 3 ESTIMATED ERROR OF FREE R VALUE : 0.017 REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN.

REMARK 3 TOTAL NUMBER OF BINS USED : 6 REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 3.10 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 3.29 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 89.9 REMARK 3 REFLECTIONS IN BIN (WORKING SET): 961 REMARK 3 BIN R VALUE (WORKING SET) : 0.293 REMARK 3 BIN FREE R VALUE : 0.403 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 4.9 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 50 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.057 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.

REMARK 3 PROTEIN ATOMS : 2492 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 5 REMARK 3 SOLVENT ATOMS : 4 REMARK 3 REMARK 3 B VALUES.

REMARK 3 FROM WILSON PLOT (A**2) : 85.2 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 71.0 REMARK 3 OVERALL ROPIC B VALUE.

REMARK 3 B11 (A**2) : 23.21 REMARK 3 B22 (A**2) : 7.23 REMARK 3 B33 (A**2) :-30.44 REMARK 3 B12 (A**2) : 0.00 REMARK 3 B13 (A**2) : 0.00 REMARK 3 B23 : 0.00 REMARK 3 REMARK 3 BULK SOLVENT MODELING.

REMARK 3 METHOD USED : FLAT MODEL REMARK 3 KSOL :0.389339 REMARK 3 BSOL :59.5295 (A**2) REMARK 3 REMARK 3 TED COORDINATE ERROR.

REMARK 3 ESD FROM LUZZATI PLOT (A) : 0.35 REMARK 3 ESD FROM SIGMAA (A) : 0.42 REMARK 3 LOW RESOLUTION CUTOFF (A) : 5.00 REMARK 3 REMARK 3 CROSS-VALIDATED ESTIMATED COORDINATE ERROR.

REMARK 3 ESD FROM C-V LUZZATI PLOT (A) : 0.51 REMARK 3 ESD FROM C-V SIGMAA (A) : 0.45 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES.

REMARK 3 BOND LENGTHS (A) : 0.008 REMARK 3 BOND ANGLES (DEGREES) : 1.2 REMARK 3 DIHEDRALANGLES (DEGREES):23.1 REMARK 3 IMPROPER ANGLES (DEGREES) : 0.73 REMARK 3 REMARK 3 ISOTROPIC THERMAL MODEL: INED REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA REMARK 3 MAIN-CHAIN BOND (A**2): 1.40; 1.50 REMARK 3 MAIN-CHAIN ANGLE (A**2): 2.49; 2.00 REMARK 3 SIDE-CHAIN BOND : 1.82; 2.00 REMARK 3 SIDE-CHAIN ANGLE (A**2): 2.93; 2.50 REMARK 3 REMARK 3 NCS MODEL : NONE REMARK 3 REMARK 3 NCS RESTRAINTS. RMS SIGMANVEIGHT REMARK 3 GROUP 1 POSITIONAL (A): NULL ;NULL REMARK 3 GROUP 1 B-FACTOR (A**2) : NULL ;NULL REMARK 3 REMARK 3 PARAMETER FILE 1 :protein_rep.param REMARK 3 PARAMETER FILE 2 :water_rep.param REMARK 3 PARAMETER FILE 3 :ion.param REMARK 3 GY FILE 1 :protein_no_cter.top REMARK 3 TOPOLOGY FILE 2 :water.top REMARK 3 TOPOLOGY FILE 4 :ion.top REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL SSBOND 1CYSS 23 CYSS 88 SSBOND 2 CYSS 22 CYSS 96 SSBOND 3 CYS S 94 CYS S 144 SSBOND 4 CYS S 99 CYS S 146 CRYST1 55.372 84.082 156.350 90.00 90.00 90.00C2221 24 ORIGX1 1.000000 0.000000 0.000000 0 ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 0.000000 0.000000 1.000000 0.00000 SCALE1 0.018060 0.000000 0.000000 0.00000 SCALE2 00 0.011893 0.000000 0.00000 SCALE3 0.000000 0.000000 0.006396 0 Figure 6 provides the dimensional structure of the XAB5 Fv complex with human lL-17A. The full homodimeric complex with exact crystallographic two-fold symmetry is W0 2014/122613 1 15 shown here. Figure 6A shows the two XAB5 Fv fragments in space-filling representation, and the |L-17A mer is shown in cartoon representation. Figure SB shows the two XAB5 Fv fragments in cartoon representation, and the |L-17A homodimer is shown in filling representation. The heavy- and light-chain of the XAB5 Fv are shown in dark and light grey, respectively. One chain of the |L-17A homodimer is shown in light grey, the other is shown in dark grey.

Figure 7 provides the three-dimensional structure of the XAB5 Fv complex with human |L-17A. Close-up view of the antibody L-CDR1 bearing the three mutations found by the structure-guided biased library approach: Asn 30, Trp 31 and Glu 32. These XAB5 side- 1O chains bute new binding interactions to the antigen human |L-17A, in particular to |L-17A residues Tyr85, Phe133, Arg124, Pro 130, Leu 49 (all from the same |L-17A subunit) and Val 45 (from the other |L-17A subunit).

Example 7. X-ray analysis of antibody variants derived by affinity tion: XAB4 The XAB4 Fv was cloned and expressed in E. coli TG1 cells with a C-terminal hexahistidine tag on the heavy-chain and a C-terminal Strep-tag on the light-chain. The recombinant protein was purified by late tography.

The XAB4 Fv fragment complex with human |L-17A was then prepared using standard methodology. In brief, human |L-17A (0.5mg) was mixed with an excess of XAB4 Fv (1.2mg) and the complex was run on a SPX-75 size-exclusion chromatography, in 10mM TRIS pH 7.4, 25mM NaCl. The protein complex was then concentrated by ultra- tion to 6.9mg/ml and crystallized.

Standard crystallization protocols were followed. In brief, crystals were grown at 19°C in VDX 24 well-plates, using the method of vapour diffusion in g drops. The protein stock was mixed 2:1 with a llization buffer containing 15% PEG 5,000 MME, 0.1M MES pH 6.5, 0.2M ammonium sulfate. Total drop size was 3.0 ul. Prior to X-ray data collection, one crystal was briefly transferred into a 1:1 mix of the llization buffer with 25% PEG 5,000 MME, 20% glycerol, and then flash cooled into liquid nitrogen.

X-ray data collection and processing was carried out using standard protocols. Briefly, X-ray data to 3.15A resolution were collected at the Swiss Light Source, beamline W0 22613 1 16 X1OSA, with a Pilatus detector, using 0.99984A X-ray radiation. In total, 720 images of 025° oscillation each were recorded at a crystal-to-detector distance of 500mm and processed with the XDS software package. The crystal belonged to space group C2221 with cell parameters a=55.76A, b=87.11A, c=156.31A, or = B = y = 90°. R-sym to 3.15A resolution was 5.5% and data completeness 99.9%.

As the crystal of the XAB4 Fv complex was nearly isomorphous with the crystal of the XAB5 Fv complex (Example 6), the structure of the latter was used as input model for ure ination by molecular ement with the program . lterative model correction and refinement was performed with Coot (Crystallographic Object- 1O Oriented Toolkit) and Autobuster version 1.11.2 (Buster version 2.11.2), until no further significant ements could be made to the crystallographic model. Final R- and R- free for all data were 0.197 and 0.253, respectively. The final refined model showed a root-mean-square deviation (RMSD) from ideal bond s and bond angles of 0.009A and 1.00, respectively. (i) Results The results of the X-ray refinement of the XAB4 Fv complex with human lL-17A are provided in Table 16 and the three-dimensional structure of this complex is shown in Figure 8. In this crystal structure, as in the XAB5 complex (Example 6), the XAB4 Fv complex has exact crystallographic 2-fold symmetry: the asymmetric unit of the crystal contains only one half of the whole, c complex. The XAB4 Fv makes contacts to both lL-17A subunits, but the vast majority of the intermolecular contacts are to only one subunit (93% of the lL-17A surface buried by one XAB4 Fv is buted by one subunit). The X-ray llography analysis confirmed that the variant antibody XAB4 retained the target specificity and bound with high affinity to essentially the same epitope as the parental XAB1 antibody. However, in the XAB4 complex structure, as in the XAB5 complex structure, the light-chain CDRL1 bears three point ons which provide enhanced binding to human lL-17A. As already described for the XAB5 complex (Example 6), Trp 31 of the XAB4 light-chain is engaged in strong hydrophobic/aromatic interactions with Tyr 85 of lL-17A and, to a lesser extent, Phe 133 of lL-17A. Asn 30 of the XAB4 light-chain donates a H-bond to the main-chain carbonyl of Pro 130 of lL-17A and is in van der Waals t to Leu 49 (same lL-17A subunit) and Val 45 (other lL- W0 2014/122613 1 17 17A subunit). Glu 32 of the XAB4 light-chain stabilizes the CDRL1 loop through olecular H-bonded interactions. Furthermore, Glu 32 makes favorable electrostatic interactions with Arg 124 of |L-17A, but is not engaged into a "head-to- head" salt-bridge interaction (Figure 9). XAB4 also differs from XAB1 in position 56 of the light-chain, as a result of an Asn to Gln mutation designed to remove a potential deamidation site. The X-ray analysis shows that Gln 56 of XAB4 makes contacts to the protein antigen residues Leu 76 and Trp 90, and reduces the solvent-accessibility of Tyr 67 and Ser 64 (Figure 10).

Table 16. X-ray refinement of the XAB4 Fv complex with |L-17A ed by the program Autobuster.

REMARK 3 REMARK 3REF|NEMENT.

REMARK 3 M : BUSTER 2.11.2 REMARK 3 AUTHORS : BRICOGNE,BLANC,BRANDL,FLENSBURG,KELLER, REMARK 3 : PACIOREK,ROVERSI,SHARFF,SMART,VONRHEIN,WOMACK; REMARK 3 : MATTHEWS,TEN RONRUD REMARK 3 REMARK 3 DATA USED IN REFINEMENT.

REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 3.15 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 78.15 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.0 REMARK 3 COMPLETENESS FOR RANGE (%) : 99.85 REMARK 3 NUMBER OF REFLECTIONS : 6881 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT.

REMARK 3 CROSS-VALIDATION METHOD : HOUT REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM REMARK 3 R VALUE (WORKING + TEST SET) : 0.1998 REMARK 3 R VALUE NG SET) : 0.1972 REMARK 3 FREE R VALUE : 0.2531 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.01 REMARK 3 FREE R VALUE TEST SET COUNT : 345 REMARK 3 ESTIMATED ERROR OF FREE R VALUE : NULL REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN.

REMARK 3 TOTAL NUMBER OF BINS USED : 5 REMARK 3 BIN RESOLUTION RANGE HIGH (ANGSTROMS) : 3.15 REMARK 3 BIN RESOLUTION RANGE LOW (ANGSTROMS) : 3.52 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 99.85 REMARK 3 REFLECTIONS IN BIN (WORKING + TEST SET) : 1916 REMARK 3 BIN R VALUE (WORKING + TEST SET) : 0.2376 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 1820 REMARK 3 BIN R VALUE NG SET) : 0.2326 REMARK 3 BIN FREE R VALUE : 0.3295 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 5.01 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 96 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : NULL REMARK 3 REMARK REMARK : 2499 REMARK NUCLEIC ACID ATOMS :0 REMARK HETEROGEN ATOMS : 5 REMARK T ATOMS : 0 REMARK REMARK B VALUES.

REMARK FROM WILSON PLOT (A**2) : 102.42 REMARK MEAN B VALUE (OVERALL, A**2) : 124.95 REMARK OVERALL ANISOTROPIC B VALUE.

REMARK B11 (A**2) : 11 REMARK B22 (A**2) : -28.0012 REMARK B33 (A**2) : 39.5523 REMARK wwe»wwwwwwwwwwwe»wwe»wwwe»wwwwwwe»wwwe»wwwe»wwwwwwwwwwwwwwwwwwwwwww NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.PROTEIN ATOMS B12 (A**2) : 0.0000 REMARK B13 (A**2) : 0.0000 REMARK B23 (A**2) : 0.0000 REMARK REMARK TED COORDINATE ERROR.

REMARK ESD FROM LUZZATI PLOT (A) : 0.787 REMARK DPI (BLOW EQ-9) BASED ON FREE R VALUE (A) : 0.474 REMARK REMARK REFERENCES: BLOW, D. (2002) ACTA CRYST D58, 792-797 REMARK REMARK CORRELATION COEFFICIENTS.

REMARK CORRELATION COEFFICIENT FO-FC : 0.9113 REMARK ATION COEFFICIENT FO-FC FREE : 0.8848 REMARK REMARK X-RAY WEIGHT: 20.89 REMARK REMARK GEOMETRY FUNCTION.

REMARK RESTRAINT LIBRARIES.

REMARK NUMBER OF LIBRARIES USED : 8 REMARK LIBRARY 1 :protgeo_eh99.dat (V1.8) 20110121 RD REMARK AMINO ACID DICTIONARY. BONDS AND ANGLES FROM REMARK ENGH AND HUBER EH99. OTHER VALUES BASED ON REMARK PREVIOUS TNT OR TAKEN FROM CCP4. INCLUDES REMARK HYDROGEN ATOMS.

REMARK LIBRARY 2 : exoticaa.dat (V1.8) 20100430 COLLECTION OF REMARK NON-STANDARD AMINO ACIDS, MAINLY EH91 WITHOUT REMARK IDEAL CE INFO REMARK LIBRARY 3 : nuclgeo.dat (V1.14) 20091104 REMARK LIBRARY 4 : l.dat (V1.15) 20080423 REMARK LIBRARY 5 : contactdat (V1.20.2.1) 20110510 REMARK LIBRARY 6 : idealdist_contact.dat (V1.7) 20110119 REMARK IDEAL-DISTANCE CONTACT TERM DATA AS USED IN REMARK PROLSQ. VALUES USED HERE ARE BASED ON THE REFMAC REMARK 5.5 IMPLEMENTATION.

REMARK LIBRARY 7 : restraints for SO4 (SULFATE ION) from cif REMARK dictionary SO4.cif using refmacdict2tnt revision REMARK 1.23.2.7; buster common-compounds v 1.0 (05 May REMARK 2011) REMARK LIBRARY 8 : assumedat (V1.10) 20110113 REMARK REMARK NUMBER OF GEOMETRIC ON TERMS DEFINED : 15 REMARK TERM COUNT WEIGHT FUNCTION.

REMARK BOND LENGTHS : 2566 ; 2.00 ; HARMONIC REMARK BOND ANGLES : 3486 ; 2.00 ; HARMONIC REMARK N ANGLES : 860; 2.00; SINUSOIDAL REMARK TRIGONAL CARBON PLANES : 61 ; 2.00; HARMONIC REMARK GENERAL PLANES : 369 ; 5.00 ; HARMONIC WO 22613 1 19 REMARK ISOTROPIC THERMAL S : 2566 ; 20.00 ; HARMONIC REMARK BAD NON-BONDED CONTACTS ' NULL ; NULL ; NULL REMARK IMPROPER TORSIONS : NULL ; NULL; NULL REMARK ROTATION ANGLES NULL; NULL; NULL REMARK CHIRAL IMPROPER TORSION 323; 5.00; SEMIHARMONIC REMARK SUM OF NCIES . NULL; NULL; NULL REMARK UTILITY DISTANCES : NULL; NULL; NULL REMARK UTILITY ANGLES ' NULL; NULL; NULL REMARK UTILITY TORSION NULL; NULL; NULL REMARK wwe»wwwwwwwwwwwe»wwe»wwwe»wwwwwwe»wwwe»wwwe»wwwwwwwwwwwwwwwwwwwwwww IDEAL-DIST CONTACT TERM 2984; 4.00; SEMIHARMONIC REMARK REMARK RMS DEVIATIONS FROM IDEAL VALUES.

REMARK BOND LENGTHS (A) : 0.009 REMARK BOND ANGLES (DEGREES) : 1.00 REMARK PEPTIDE OMEGA TORSION ANGLES (DEGREES) : 4.39 REMARK OTHER TORSION ANGLES (DEGREES) : 18.96 REMARK REMARK SIMILARITY.

REMARK NCS.

REMARK NCS REPRESENTATION : NONE REMARK TARGET RESTRAINTS.

REMARK TARGET REPRESENTATION : LSSR REMARK TARGET STRUCTURE : xab5_i|17a_complex_fina|_buster.pdb REMARK REMARK TLS DETAILS.

REMARK NUMBER OF TLS GROUPS : 3 REMARK REMARK TLS GROUP: 1 REMARK SET : { H|*} REMARK ORIGIN FOR THE GROUP (A): 10.9676 6 -10.1379 REMARK T TENSOR REMARK T11: -0.1266 T22: 0.0257 REMARK T33: 9 T12: -0.3040 REMARK T13: -0.0312 T23: 0.1050 REMARK L TENSOR REMARK L11: 7.4496 L22: 4.4770 REMARK L33: 4.2880 L12: 1.1123 REMARK L13: -1.8044 L23: 3.0307 REMARK S TENSOR REMARK S11: 0.2013 S12: 0.3070 S13: -0.5774 REMARK S21: 0.4752 S22: -0.5377 S23: 0.7096 REMARK S31: 1.0885 S32: -1.0885 S33: 0.3364 REMARK REMARK TLS GROUP: 2 REMARK SET : { ||*} REMARK ORIGIN FOR THE GROUP (A): 22.7365 0.7101 -35.1243 REMARK T TENSOR REMARK T11: -0.1883 T22: 0.1529 REMARK T33: -0.3560 T12: 0.0318 REMARK T13: -0.1985 T23: 0.0144 REMARK L TENSOR REMARK L11: 2.7494 L22: 9.3427 REMARK L33: 3.8648 L12: 0.8073 REMARK L13: -0.6650 L23: -2.0544 REMARK S TENSOR REMARK S11: 0.0485 S12: 0.3188 S13: 0.0579 REMARK S21: 0.0595 S22: 0.1433 S23: 0.7000 REMARK S31: 0.0050 S32: -0.6066 S33: -0.1917 REMARK REMARK TLS GROUP : 3 REMARK 3 SET : { L|*} REMARK 3 ORIGIN FOR THE GROUP (A): 33.2517 -11.1794 -14.2151 REMARK 3 T TENSOR REMARK 3 T11: 0.0667 T22: -0.1645 REMARK 3 T33: -0.2360 T12: 0.1870 REMARK 3 T13: -0.2270 T23: -0.1209 REMARK 3 L TENSOR REMARK 3 L11: 3.3694 L22: 3.7848 REMARK 3 L33: 8.8916 L12: -0.6497 REMARK 3 L13: -2.6132 L23: 0.8234 REMARK 3 S TENSOR REMARK 3 S11: -0.0839 S12: -0.2629 S13: -0.1560 REMARK 3 S21: 0.3804 S22: 0.7574 S23: 8 REMARK 3 S31: 1.0885 S32: 1.0885 S33: -0.6736 REMARK 3 REMARK 3 REFINEMENT NOTES.

REMARK 3 NUMBER OF MENT NOTES : 1 REMARK 3 NOTE 1 : IDEAL-DIST T TERM CONTACT SETUP. ALL ATOMS REMARK 3 HAVE CCP4 ATOM TYPE FROM LIBRARY REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL REMARK 3 SSBOND 1 CYS H 22 CYS H 96 1555 1555 2.03 SSBOND 2 CYS I 94 CYS I 144 1555 1555 2.05 SSBOND 3 CYS I 99 CYS I 146 1555 1555 2.04 SSBOND 4 CYS L 23 CYS L 88 1555 1555 2.07 CISPEP 1 TYR I 85 PRO I 86 0 3.67 CISPEP 2 GLU I 125 PRO I 126 0 -9.25 CISPEP 3 PRO I 126 PRO I 127 0 5.92 CISPEP 4 SER L 7 PRO L 8 0 -6.50 CISPEP 5 TYR L 94 PRO L 95 0 -6.52 CRYST1 55.760 87.109 156.306 90.00 90.00 90.00 C 2 2 21 Figure 8 provides the three-dimensional structure of the XAB4 Fv complex with human lL-17A. Figure 8A shows the two XAB4 Fv fragments in space-filling representation, and the lL-17A homodimer is shown in cartoon representation. Figure 8B shows the two XAB4 Fv fragments in cartoon representation, and the lL-17A homodimer is shown in space-filling representation. The heavy- and light-chain of the XAB4 Fv are shown in dark and light grey, respectively. One chain of the lL-17A homodimer is shown in light grey, the other is shown in dark grey.

Figure 9 provides the dimensional structure of the XAB4 Fv complex with human lL-17A as a close-up view of the antibody L-CDR1 g the three mutations found by the structure-guided biased library approach: Asn 30, Trp 31 and Glu 32. These XAB4 side-chains contribute new binding interactions to the antigen human lL-17A, in particular to lL-17A residues Tyr85, Phe133, Arg124, Pro 130, Leu 49 (all from the same lL-17A t) and Val 45 (from the other lL-17A subunit).

W0 2014/122613 1 21 Figure 10 provides the dimensional structure of the XAB4 Fv complex with human lL-17A as a close-up view of the dy L-CDR2 showing the Asn 56 to Gln mutation.

This XAB4 side-chain contributes binding contacts to lL-17A residues Trp 90 and Leu 76, and reduces the solvent-accessibility of Tyr 67 and Ser 64 (all from the same lL-17A To summarize, X-ray crystallography analysis confirmed that the variant antibodies selected for further analysis retained their target specificity and bound with high ty to essentially the same epitope as the parental XA81 antibody. Tighter binding between each of the variant antibodies and lL-17A was observed, as a result of additional or improved binding contacts (see Table 17 below). r characterisation of the variant antibodies was conducted as bed below.

Table 17. X-ray analyses of the |L-17A epitope bound by XAB1, XABZ, XAB4 and XABS: summary and structure-based, qualitative classification of epitope residues. (*): residue contributed by the second |L-17A subunit.

Epitope XAB1 XAB2 XAB4 XAB5 residue class Very important Arg 78, Glu Arg 78, Glu Arg 78, Glu Arg 78, Glu epitope 80, Trp 90 80, Trp 9O 80, Tyr 85, Trp 80, Tyr 85, Trp residues 90, Arg 124 90, Arg 124 Other Pro 82, Ser Arg 43*, Pro Pro 82, Ser Pro 82, Ser ant 87, Val 88, 82, Ser 87, 87, Val 88 87, Val 88 epitope Arg 124 Val 88, Arg residues 124 Additional Val 45*, Leu Pro 42*, Val Val 45*, Leu Val 45*, Leu contributions 49, He 51, Asp 45*, Leu 49, 49, Asp 81, 49, Asp 81, 81, Glu 83, He 51, Asp 81, Glu 83, Pro Glu 83, Pro Tyr 85, Asn Glu 83, Tyr 86, Pro 130, 86, Pro 130, 131, Lys 137* 85, Asn 131, Phe 133, Lys Phe 133, Lys Lys 137* 137* 137* Little or no Thr 44*, Leu Leu 76, His Arg 43*, Asn Arg 43*, Asn direct 76, His 77, 77, Asn 79, 50, Ser 64, 50, Leu 76, contribution Asn 79, Arg Arg 84, Pro Tyr 67, Leu His 77, Asn 84, Pro 86, 86, Lys 93, 76, His 77, 79, Arg 84, Lys 93, Glu Glu 118*, Pro Asn 79, Arg Glu 118*, Leu 118*, Pro 130, 130, Phe 133 84, Glu 118*, 122, Asn 131, Phe 133 Leu 122, Asn Leu 135* 131, Leu 135* Example 8. Affinity measurements and cross-reactivity measured by BiacoreTM Determination of kinetic binding ters was achieved by surface plasmon resonance measurements using the optical biosensor BiacoreTM T200 or T100 W0 2014/122613 1 23 //www.biacore.com). This technology allows the label-free determination of the microscopic rate constants for binding (ka) and dissociation (kd) of a ligand to a receptor.

It is therefore especially suited for characterizing the antibody-antigen interactions. ct binding of antibodies to the eTM chip surface was done via an anti-human lg antibody (GE care Bio-Sciences AB; Cat.No. BR39) 25 ug/ml in lization buffer (10mM Sodium acetate pH 5.0) or through protein A (RepliGen: rPA-50) 20 ug/ml in immobilization buffer (10mM Sodium acetate pH 5.0 or pH 4.0).

Antibody was diluted into blank buffer to a final concentration of 1.00 or 1.25 ug/ml.

Affinity measurements for the determination of dissociation constants of XAB4 or XAB1 1O was performed for recombinant hulL-17A (SEQ ID NO: 78, e.g. 2-fold increasing concentrations from 0.14 to 8.8 nM), recombinant hulL-17A/F heterodimer (e.g. 2-fold sing concentrations from 0.13 to 8 nM), recombinant 7F (SEQ ID NO: 77; e.g. 2-fold increasing concentrations from 7.8 to 500 nM) cynomolgus IL-17A (SEQ ID NO: 79; e.g. 2-fold increasing concentrations from 0.63 to 40 nM) rhesus IL-17A (SEQ ID NO: 82; e.g. 2-fold increasing concentrations from 1.6 to 100 nM), marmoset IL-17A (SEQ ID NO: 82; e.g. 2-fold increasing concentrations from 0.63 to 40 nM), recombinant A (SEQ ID NO: 83; e.g. 2-fold increasing concentrations from 0.78 to 50 nM), recombinant mlL-17A/F (R&D Systems® Cat# 5390-IL; e.g. 2-fold increasing trations from 1.25 to 40 nM) rat IL-17A (SEQ ID NO: 85; e.g. 2-fold increasing concentrations from 0.78 to 50 nM), using the indirect coupling/binding method (see above) and surface was regenerated with 10 mM glycine pH 1.75 or MgClz (3 M). One chip surface was coated and reused t significant loss of g capacity. Ligand concentrations were chosen to start below the KB and to end at a concentration higher than ten times the KB.

Similar but not identical conditions were used to measure affinity of XAB2 and XAB3.

The kinetic traces were evaluated with the BiacoreTM T200 Control Software version 1.0. The full set of these traces with increasing concentrations is taken together and is called a run. Two zero concentration samples (blank runs) were included in each analyte concentration series to allow double-referencing during data evaluation W0 2014/122613 1 24 Results The g of the anti-IL-17 antibodies XAB4, XABl, XABZ and XABB to human, cynomolgus monkey, marmoset monkey, rhesus monkey, mouse and rat |L-17A, to human and mouse /F heterodimer and to human |L-17F was determined by surface plasmon resonance using the BiacoreTM technology.

The kinetic rate constants for association (ka) and dissociation (kd), as well as the dissociation equilibrium constant (KD) were calculated.

The affinity data of XAB4 is shown in Table 18, the affinity data of XA81 is shown in 1O Table 19, the affinity data of XABZ is shown in Table 20, and the affinity data of XABB is shown in Table 21. Affinity maturation of XABl, XABZ and XABB increased the affinity towards human, cynomolgus monkey, mouse and rat .

Table 18. Affinity and kinetic rate constants of XAB4 binding. ka (1/Ms) kd(1/s) KD (M) huIL-17A 5.7 i 0.0 E-12 huIL-17A/F < 1.1 i 0.0 E-11* -17A 3.1 i 0.4 E-11 marmIL-17A 1.2 i 0.0 E+O6 2.2 i 0.0 E-05 1.8 i 0.0 E-11 rhele-17A 4.0 i 0.1 E-11 m|L-17A 1.6: 0.1 E-10 m|L-17A/F 2.604 E-10 ratIL-17A 8.4 i 1.0 E-11 n.d. = not determinable, applied antigen conc. range too low and non-specific binding of antigen to reference flow cell ed at the highest antigen concentrations (500-50 I0|V|)- ciation rate outside the limits that can be measured by the instrument (kd < 1 x 10' 1/s) W0 2014/122613 1 25 Table 19. Affinity and c rate constants of XAB1 binding. —huIL-17A 2.33E+06 9.39E-05 4.03E-11 —huIL-17A/F 9.097E+05 0.001342 1.475E-09 n.d. = not determinable, applied antigen conc. range too low and non-specific binding of antigen to reference flow cell observed at three highest n trations (500-50 pM).

Table 20. Affinity and kinetic rate constants of XABZ binding. huIL-17A 4.09E+06 7.12E-05 1.76E-11 Table 21. ty and kinetic rate constants of XAB3 binding. huIL-17A 5.48E+06 5.01E-05 9.58E-12 huIL-17A/F 3.37E+06 1.03E-04 3.29E-11 cynolL-17A 1.21 E+06 4.23E-05 3.49E-11 mIL-17A 5.87E+05 1.01 E-04 1.74E-10 ratIL-17A 9.05E+05 7.59E-05 8.26E-11 n.d. = not determinable W0 22613 1 26 The affinities and kinetic rate constants for XAB2, XAB3 and XAB5 are comparable to those observed for XAB4.

Example 9. Binding in ELISA to IL-17A and other family members A titration of the antibodies of interest on different antigens was carried out. Briefly, wells of ELISA microtiter plates (Nunc Immuno plates MaxiSorp: Invitrogen, Cat# 4- 39454A) were coated with 1 ug/ml of recombinant hulL-17A (SEQ ID NO: 76; 1.8 mg/ml), recombinant hulL-17A/F (0.59 mg/ml), recombinant hulL-17F (SEQ ID NO: 77; 1.8 mg/ml)), recombinant hulL-17B (R&D Systems® Cat# 1248|B/CF), recombinant 1O hulL-17C (R&D Systems® Cat# 1234iL/CF), recombinant hulL-17D (R&D Systems® Cat# /CF), recombinant hulL-17E (R&D Systems® Cat# L/CF), recombinant -17A (SEQ ID NO: 79; 0.21 , recombinant cynolL-17F (SEQ ID NO: 80; 1.525 mg/ml), recombinant mlL-17A (SEQ ID NO: 83; 2.8 mg/ml), inant mlL-17A/F (R&D Systems® Cat# 5390-IL), inant mlL-17F (SEQ ID NO: 84; 0.2 mg/ml) and recombinant ratlL-17A (SEQ ID NO: 85; 3.8 mg/ml) (100 ul/well) in ate buffered saline (PBS) without Ca and Mg (10x; Invitrogen Cat# 14200-083) 0.02% NaN3 (Sigma Cat# S-8032) and incubated overnight at 4°C.

The following day, microtiter plates were blocked with 300 pl of PBS/2% BSA (fraction V; Roche Cat# 10 735 094 001)/0.02% NaN3 for 1 h at 37°C. Plates were then washed 4 times with PBS/0.05% Tween 20 (Sigma Cat# P7949) /0.02% NaN3. XAB4 or XAB1 were added at 1 ug/ml in triplicate wells (100 ul/well) for 3 h at room temperature.

To verify coating of antigens to the plates, control antibodies were used and in particular, a mouse mAb anti-hulL-17F, (Novartis, 5 ug/ml) a goat anti-hu-IL-17B (R&D s® Cat# AF1248; 10 ug/ml), a mouse mAb anti-hulL-17C (R&D Systems® Cat# MAB1234; 10 ug/ml), a goat anti-hulL-17D (R&D Systems® Cat# AF1504; 10 ug/ml), a mouse mAb anti hu-IL-17E (R&D Systems® Cat# 8; 10 ug/ml), a mouse anti- mlL-17A or anti-mlL-17A/F (Novartis; 1 ug/ml), and a rat lL-17F (R&D Systems® Cat# MAB2057; 1 ug/ml; ) (100 l in PBS, 0.02% NaN3 for 3h at RT).

Plates were then washed 4 times with PBS/0.05% Tween 20/0.02% NaN3. Then, an alkaline phosphatase-conjugated goat anti-human IgG antibody (Sigma Cat# A9544) was added to the wells that received test antibody at a dilution of 1/20000 (100 ul/well) W0 2014/122613 1 27 for 2h 30min at RT. To the wells, that received mouse mAb, an alkaline phosphatase- conjugated goat anti-mouse lgG antibody (Sigma Cat# A7434) was added at a dilution of 1/1000O (1OO ul/well) for 2 h 30 min at RT. An alkaline phosphatase conjugated mouse anti goat lgG antibody (Sigma Cat# A8062) was added to the goat antibodies at a dilution of 1/5000O (1OO l) for 2 h 30 min at RT. Plates were then washed 4 times and 100 pl of the substrate (p-nitrophenyl phosphate tablets; Sigma; 5 mg Cat# N9389; 20 mg Cat#. N2765) dissolved in diethanolamine buffer pH 9.8, to give a final tration of 1 mg/ml, were added to each well.

Plates were read after 30 min in a Spectra Max M5 Microplate Reader (Molecular Devices) using filters of 405 and 490 nm. Values are the means 1r SEM of triplicate values.

Results These s show that XAB4 and XAB1 are able to bind human and mouse |L-17A, and human and mouse |L-17A/F. In addition it is shown that XAB4 is able to bind cynomolgus and rat |L-17A. Binding to human, lgus and mouse |L-17F was not detected under these experimental conditions as well as binding to other human family members B, lL-17C, lL-17D and lL-17E).

Table 22. Cross-reactivity of XAB4 and XAB1 to human, cynomolgus monkey, mouse and rat |L-17 family members, by ELISA.

XAB4 Control XAB1 Control (1 ug/ml) antibody (1 or (1 ug/ml) antibody (1 or 0D values 10 ug/ml) O.D values 10 ug/ml) (mean 1r SEM) O.D values (mean 1r O.D values (mean 1r SEM) (mean + SEM) hu lL-17A 2471 i 0.0448 - hu lL-17A/F 2137 i 0.0429 - hu |L-17F 0.049 i 0.0056 1.913 + 0. 0483 W0 2014/122613 1 28 hu |L-17B ---0.034 i 0.0007 0.283 i 0.0066 0.049 1.441 i 0.0283 hu |L-17C ---0.036 i 0.0002 0.290 i 0.0027 0.032 0.558 i 0.0169 hu |L-17D ---0.034 i 0.0005 0.292 i 0.0048 0.031 0.867 i 0.0372 hu |L-17E 0.035 i 0.0014 0.833 i 0.0239 0.033 2.054 i 0.0378 i0.0003 mouse |L-17A---1.585 i 0.0428 1.086 i 0.0119 1.439 3.697 i 0.0602 mouse 2.263 i 0.0243 1.142 i 0.0315 1.762 2.084 i 0.0223 mouse |L-17F---0.098 i 0.0060 1.294 i 0.0134 0.044 1.770 i 0.0302 Example 10. reactivity to other human, mouse and rat eukins by ELISA In another set of experiments the cross-reactivity of antibodies of the disclosure for selected human, mouse or rat cytokines was evaluated.

Triplicate wells of ELISA microtiter plates (Nunc Immuno plates MaxiSorp: lnvitrogen Cat# 4-39454A) were coated with 100 pl/well of the following cytokines: recombinant hulL1B (Novartis), recombinant hulL-3 (R&D Systems® Cat# 203-IL/CF), recombinant hulL-4 (R&D Systems® Cat# 204-IL/CF), recombinant hulL-6 (R&D s® Cat# 206-IL-1010/CF), recombinant hulL-8 (R&D s® Cat# 208-IL-O10/CF), recombinant hulL-12 (R&D s® Cat# 219-IL-OO5/CF), recombinant hulL-13 (Novartis), recombinant hulL-17A (SEQ ID NO: 76), recombinant hulL-17A/F, recombinant hulL-17F (SEQ ID NO: 77), recombinant hulL-18 (MBL Cat# ), recombinant hulL-20 (Novartis), recombinant hulL-23 (R&D Systems® Cat# 1290-IL- W0 2014/122613 1 29 O10/CF), recombinant hulFNy (Roche), inant huTNFd tis), recombinant huEGF (Sigma Cat#E9644.), recombinant Z tis), recombinant m|L-1B (R&D Systems® Cat# 401-ML), recombinant mlL-2 (R&D Systems® -020/CF), recombinant mlL-6 (R&D Systems® Cat# 406-ML-O10/CF), recombinant mlL-12 (R&D Systems® Cat# 419-ML-O10/CF), inant mlL-17A (SEQ ID NO: 83), recombinant mlL-17A/F (R&D s® Cat# 5390-IL), recombinant mlL-17F (R&D Systems® Cat# 2057—lL/CF), recombinant mlL-18 (MBL Cat#BOO4-5), recombinant mlL-23 (R&D Systems® Cat# 1887-ML), recombinant mlFN-y (R&D Systems® Cat# 485-MT), recombinant mTNFd (R&D Systems® Cat# 410-MT), recombinant rat |L-4 (R&D 1O Systems® Cat# 504-RL/CF), recombinant rat lL-6 (R&D Systems® Cat# 506-RL-O10), recombinant ratlL-12 (R&D Systems® Cat# 1760-RL/CF), recombinant ratlL-17A (SEQ ID NO: 85), recombinant 23 (R&D Systems® Cat# 3136-RL-O10/CF), recombinant d (R&D Systems® Cat# 510-RT/CF), at 1 ug/ml with the exception of recombinant mlL-6, recombinant mlL-12 and recombinant mTNFd which were coated at 0.5 ug/ml in phosphate buffered saline (PBS) without Ca and Mg (10x; Invitrogen Cat# 14200-083) 0.02% NaN3 (Sigma Cat# S-8032) and incubated overnight at 4°C.

The following day, microtiter plates were d with 300 pl of PBS/2% BSA (fraction V; Roche Cat# 10 735 094 OO1)/0.02% NaN3 for 1 h at 37°C. Plates were then washed 4 times with 05% Tween 20 (Sigma Cat# P7949) /0.02% NaN3.

The antibodies of the disclosure were added at 10 ug/ml (1OO ul/well) for 3 h at room temperature. To verify coating of antigens to the plates, 1OO ul/well of the ing control antibodies were used: a mouse anti-hulL1B (R&D Systems® Cat# MABSO1), a mouse anti-hulL-3 (R&D Systems® Cat# MABSOS), a mouse anti-hulL4 (R&D Systems® Cat# MABSO4), a mouse anti-hulL-6 (R&D Systems® Cat# MABZOS), a mouse anti-hu-IL8 (R&D Systems® Cat# MABZO8), a mouse anti-hulL-12 (R&D Systems® Cat# MABZ19), a mouse anti-hulL-13 (Novartis), a mouse anti-hulL-17A (Novartis), a mouse anti-hulL-17F (Novartis), a mouse anti-hulL-18 (MBL Cat# DO43-3), a mouse anti-hulL-20 (Abcam Cat# ab57227), a goat anti-hulL-23 (R&D Systems® Cat# AF1716), a mouse anti-hulFN-y (R&D Systems® Cat# MABZ85), a mouse anti- huTNF-d (R&D Systems® Cat# MABS10), a mouse u-EGF (R&D s® Cat# MABZBS), a human anti-huTGFBZ (Novartis), a rat anti-m|L-1B (R&D Systems® Cat# MAB401), a rat anti-mlL-2 (R&D Systems® Cat# MAB402), a rat anti-mlL-6 (R&D W0 2014/122613 1 30 Systems® Cat# ), a rat lL-12 (R&D s® Cat# ), a mouse anti-m/ratlL-17A (Novartis), a rat anti-mlL-17F (R&D Systems® Cat# MABZO57), a rat anti-mlL-18 (MBL Cat# D047-3), a rat anti-mlFN-y (R&D Systems® Cat# MAB485), a goat anti-mTNFd (R&D Systems® Cat# AFNA), a mouse anti-rat |L-4 (R&D Systems® Cat# MAB504), a goat anti-rat |L-6 (R&D Systems® Cat# AF506), a goat anti-rat |L-12 (R&D Systems® Cat# AF1760), a mouse anti-rat |L-23 (R&D Systems® Cat# MABS510), a mouse anti-rat TNFd (R&D Systems® Cat# MAB510). They were added at 1 or 5 ug/ml, in PBS, 0.02% NaN3 for 3 h at RT.

Plates were then washed 4 times with PBS/0.05% Tween 20/0.02% NaN3. Then, an 1O alkaline phosphatase-conjugated goat anti-human lgG antibody (Sigma Cat# A9544) was added to the wells with human antibodies at a dilution of 1/20000 (100 ul/well). An alkaline phosphatase-conjugated goat ouse lgG antibody (Sigma Cat# A1047) was added to the wells with mouse antibodies at a dilution of 1/10000 (100 ul/well). An alkaline phosphatase-conjugated rabbit anti-goat lgG antibody (Sigma Cat# A7650) was added to the wells with goat antibodies at a dilution of 1/1000 (100 ul/well) and an alkaline phosphatase-conjugated rabbit anti rat-lgG antibody (Sigma Cat# A6066) was added to the wells with rat antibodies at a dilution of 0 (100 l). The secondary antibodies were incubated for 2 h 30 min at RT. Plates were then washed 4 times and 100 pl of the substrate (p-nitrophenyl phosphate tablets; Sigma; 5 mg Cat#.

N9389 or 20 mg Cat# N2765) dissolved in diethanolamine buffer pH 9.8, to give a final concentration of 1 mg/ml, were added to each well.

Plates were read after 30 min at RT or ON at 40C in a Spectra Max M5 late Reader (Molecular Devices) using filters of 405 and 490 nm. Values are the means 1r SEM of triplicate values.

Results The data obtained show that both XAB4 and XAB1 are highly selective for |L-17A of human, mouse and rat origin and for |L-17A/F of human and mouse origin. In addition, under the conditions , the reactivity of XAB1 at 10 ug/ml for human |L-17F (not seen at 1 ug/ml, see above) is not observed with XAB4. Reactivity for the other cytokines tested was not detected.

W0 2014/122613 1 31 Table 23. Cross-reactivity of XAB4 and XAB1 to human cytokines by ELISA.

XAB4 l antibody XA81 Control antibody (10 ug/ml) (5 ug/ml) (10 ug/ml) (1 ug/ml) O.D values O.D values O.D values O.D values (mean 1r SEM) (mean 1r SEM) (mean 1r SEM) (mean 1r SEM) |L1B 0.015 1r 0.0075 0.867 1r 0.0107 -0.110 1r 0.0901 3.071 1r 0.0486 NB. the negative values are due to the fact that the blank (O.D. value of wells without specific antibodies) is subtracted.

Table 24. Cross-reactivity of XAB4 and XAB1 to mouse cytokines by ELISA.

XAB4 Control antibody XA81 Control antibody (10 ug/ml) (5 ug/ml) (10 ug/ml) (5 ug/ml) O.D values O.D values O.D values O.D values (mean 1r SEM) (mean 1r SEM ) (mean 1r SEM ) (mean 1r SEM ) lL-1B 0 022 i 0.0057 0611 i 0.0665 0.007 i 0.0123 0.624 i 0.0455 W0 2014/122613 1 32 |L17F 0.034 i 0.0122 3.359 i 0.0247 -0.058 i 0.0326 3.264 i 0.0309 NB. the negative values are due to the fact that the blank (O.D. value of wells without specific antibodies) is cted.

Table 25. Cross-reactivity of XAB4 and XAB1 to rat cytokines by ELISA.

XAB4 Control antibody XAB1 Control antibody (10 ug/ml) (5 ug/ml) (10 ug/ml) (5 ug/ml) O.D values O.D values O.D values O.D values (mean 1r SEM) (mean 1r SEM) (mean 1r SEM) (mean 1r SEM) lL4 0.026 1r 0.0082 3.168 1r 0.0297 0.017 1r 0.0092 3.324 1r 0.1092 |L6 0.021 i 0.0028 3.116 i 0.0318 0.000 i 0.0141 3.253 i 0.1078 NB. the negative values are due to the fact that the blank (O.D. value of wells without specific antibodies) is subtracted.

W0 2014/122613 1 33 Example 11. IL-17A — IL-17RA and IL-17A/F-IL-17RA in Vitro itive binding inhibition assay Human lL-17RA was used from a stock solution (BTP22599: 1.68 mg/ml = 46.2 uM).

ELISA microtiter plates were coated with human lL-17RA (100 ul/well, 1 ug/ml, ~27.5 nM) in PBS/ 0.02% NaN3 and incubated overnight at room temperature. The following day the plates were blocked with 300 pl of PBS/2% BSA/0.02% NaN3 for 1 h at 37°C.

Then the plates were washed 4 times with PBS/0.05% Tween20/0.02% NaN3.

Following this preparation, titration of antibody variants (50 ul, concentrations from 12 nM to 0.12 nM for lL-17A and 1200 nM to 40 nM for lL-17A/F, steps of 3) were pre- 1O ted with human lL-17A biotin (50 ul at 0.94 nM) or lL-17A/F (50 ul at 31 nM) for minutes at room temperature. 100 pl of the mixture were added to the well for 3 hours and 30 minutes at room temperature. After washing with PBS/0.05% Tween20/0.02% NaN3, four times alkaline phosphatase-conjugated streptavidin was added at a final dilution of 1/10000 (100 ul/ well). After 45 minutes at room ature plates were washed again 4 times with PBS/0.05% Tween20/0.02% NaN3 and the ate p-nitrophenylphosphate in diethanolamine buffer pH 9.8 (1 mg/ml), was added (100 ul/well).

Plates were read after 30 minutes in spectra Max M5 Microplate reader, filters 405 and 490nm (triplicates). The calculation of the percentage of inhibition and |C5o for different antibody variants was done using a four parameter ic model (Excel leit; FlT model 205).

Results Data show that both XAB4 and XAB1 are able to block the binding of hulL-17A and hulL-17A/F to the hulL-17RA. The higher affinity of XAB4 for lL-17A and lL-17A/F is reflected in a higher inhibitory capacity. |C5o values are ed in the table. The higher concentrations needed to block the lL-17A/F-lL-17RA interaction are mostly explained by the fact that about 30 fold higher trations of lL-17A/F were used in the assay.

The dy binds to the A subunit of NF and ore cannot prevent binding of the F subunit to the lL-17RA. However, binding of F to lL-17RA is rather weak, in the 300nM range.

W0 2014/122613 2014/058854 1 34 Table 26. XAB4 and XAB1 t the binding of hulL-17A and hulL-17A/F to hulL- 17RA.

Ligand\Receptor XAB4 XAB1 Control interaction |C50 (nM) (mean 1r |C50 (nM) (mean antibody (nM) SEM) 1r SEM) hulL-17A\hu|L-17RA 0.321 i 0.037 0.830 i 0.112 >60 7A/F\hu|L-17RA 153.9 i 18.9 301.3 i 51.9 _ Example 12. In Vitro neutralisation of human IL-17A and IL-17A/F activity by antibody variants of the disclosure (i) Assay on /6 cells (human chondrocyte cell line) I6, or C-20/A4, clone 6, (Goldring MB, et al 1994, J Clin Invest; 94:2307-16) cells were cultured in RPMI (Gibco Cat# 61870-010) supplemented with 10% fetal calf 1O serum low lgG (Gibco Cat# 16250-078; lot 1074403), B-mercapto ethanol (5x10'5 M final), and Normocin (0.1 mg/ml; InvivoGen Cat# ant-nr-2).

The cells were detached from plastic using an Accutase solution (PAA Cat# L11-007).

Cells were distributed into 96 well microtiter plates at a density of 5x103 in 100u| well in RPMI 1640 (Gibco Cat# 61870-010) without fetal calf serum, B-mercaptoethanol (5x10'5 M final) and Normocin (0.1 mg/ml).

The C20A4CI6 cells were allowed to adhere to the plates overnight. The next morning, different concentrations of recombinant 7A (SEQ ID NO: 76; MW 32000), recombinant hulL-17A/F (MW 32800), recombinant hulL-17F (SEQ ID NO: 77; MW 30000), or control medium in the presence of human TNFd (Novartis; MW 17500) were added in a volume of 50 ul to triplicate wells in the presence of 50 ul of different concentrations of test antibody (XAB4; XAB1), control antibody (Simulect® 1.1% solution, Batch C0011; 831179) or control medium to reach the final volume of 200 ul/well and the final concentration of 0.5% fetal calf serum.

W0 2014/122613 1 35 HulL-17A (30 pM), hulL-17A/F (300 pM) and hulL-17F (10 nM) were added together with huTNFd (6pM). XAB4 (MW 150000) was added in a concentration range from 1 to 0.003 nM to neutralize hulL-17A, in a concentration range from 10 to 0.03 nM to neutralize hulL-17A/F and in a concentration range from 3 uM to 30 nM for hulL-17F.

XAB1 (MW 150000) was added in a concentration range from 3 to 0.01 nM to neutralize hulL-17A, in a concentration range from 10 to 0.03 nM to neutralize hulL-17A/F and in a concentration range from 3 uM to 30 nM for hulL-17F. Simulect® was added in a concentration range between 3 uM to 100 nM. Culture supernatants were collected after an incubation of 24 h and hulL-6 production was measured by ELISA. 1O (ii) Assay on BJ cells (human fibroblasts) BJ cells (human skin fibroblasts from ATCC Cat# CRL 2522) were cultured in RPMI (Gibco Cat# 61870-010) supplemented with 10% fetal calf serum ultra-low lgG (Gibco Cat# 16250-078; lot 1074403), aptoethanol (5x10'5 M final) and Normocin (0.1 mg/ml; Gen Cat# ant-nr-2). The cells were detached from c using an Accutase solution (PAA Cat# L11-007).

The cells were distributed into 96 well microtiter plates at a density of 5x103 in 100 pl well in RPMI 1640 t fetal calf serum, B-mercaptoethanol (5x10'5 M final) and Normocin (0.1 mg/ml). The BJ cells were allowed to adhere to the plates overnight. The next morning, different concentrations of rhulL-17A (SEQ ID NO: 76; MW , rhulL- 17A/F (MW 32800) and rhulL-17F (SEQ ID NO: 77; MW 30000), or l medium in the presence of human TNFd (Novartis; MW 17500) were added in a volume of 50 ul to triplicate wells in the presence of 50 ul of different concentrations of test antibody (XAB4; XAB1), l antibody (Simulect® 1.1% solution, Batch # C0011; 831179), or control medium to reach the final volume of 200 ul/well and the final concentration of 2.5% fetal calf serum.

HulL-17A (30 pM), hulL-17A/F (300 pM) and hulL-17F (10 nM) were added together with huTNFd (6 pM). XAB4 (MW ) was added in a concentration range from 1 to 0.003 nM to neutralize hulL-17A, in a concentration range from 10 to 0.03 nM to lize hulL-17A/F and in a tration range from 3 uM to 30 nM for hulL-17F.

XAB1 (MW 150000) was added in a concentration range from 3 to 0.01 nM to neutralize hulL-17A, in a concentration range from 10 to 0.03 nM to neutralize hulL-17A/F and in a W0 2014/122613 1 36 concentration range from 3 uM to 30 nM for 7F. ct® was added in a concentration range between 3 uM to 100 nM. Culture supernatants were collected after an incubation of 24 h and hulL-6 and huGROd production were measured by ELISA. (iii) Detection assays 1) ELISA for detection of human IL-6 production ELISA microtiter plates were coated with an anti-human lL-6 mouse Mab (R&D Systems® Cat# MABZOS; 100ul/well at 1ug/ml) in PBS 0.02%NaN3 and incubated overnight at +4°C.The following day, microtiter plates were blocked with 300ul of PBS/2% BSA/0.02% NaN3 for 3h at room temperature. Plates were then washed 4 1O times with PBS/0.05%Tween20/0.02% NaN3. Culture supernatants of CZOA4C|6 (final dilution 1:5 for cultures stimulated with hulL-17A plus huTNFd, or 1:2 for cultures stimulated with huTNFd plus hulL-17A/F or lL-17F; 100ul/well) or BJ cells (final dilution 1:10 for cultures stimulated with hulL-17A plus huTNFd, or 1:5 for cultures ated with huTNFd plus hulL-17A/F or lL-17F; 1OO ul/well) were added.

To establish a titration curve, rhulL-6 (Novartis; 100ul/well) was titrated from 500 pg/ml to 7.8 pg/ml in 1:2 dilution steps. After an overnight incubation at room temperature, plates were washed 4 times with PBS/0.05% Tween 2% NaN3. A - conjugated goat anti-human lL-6 antibody was added (R&D Systems® Cat# BAF206; ng/ml; 1OO ul/well). Samples were left to react for 4 h at room temperature. After washing (4 , ne phosphatase-conjugated avidin (Jackson lmmunoresearch Cat# 016084) was added at a final dilution of 1/1000O (1OO ul/well).

After 40 minutes at room temperature, plates were washed again 4 times. P-Nitrophenyl Phosphate substrate tablets (Sigma; 5 mg, Cat# N9389; 20 mg, Cat# N2765) were dissolved in nolamine buffer pH 9.8 to give a final concentration of 1 mg/ml. 100 pl were added to each well and the CD. was read after 1 h in a Spectra Max M5 Microplate Reader (Molecular Devices) using filters of 405 and 490 nm.

W0 2014/122613 1 37 2) ELISA for detection of human GROa production ELISA microtiter plates were coated with an anti-human GROd mouse mAb (R&D Systems® Systems® Cat# MABZ75; 1OO ul/well at 1.5 ug/ml) in PBS/0.02% NaN3 and incubated overnight at 4°C.The following day, microtiter plates were blocked with 300 pl of PBS/2% BSA/0.02% NaN3 for 3 h at room temperature. Plates were then washed 4 times with PBS/0.05% Tween20/0.02% NaN3. e atants of BJ cells (final dilution 1:2; 100ul/well) were added.

To establish a titration curve, human GROd (R&D Systems® Cat# /CF; 1OO ul/well) was titrated from 2 ng/ml to 0.03 ng/ml in 1:2 dilution steps.) After an overnight 1O incubation at room temperature, plates were washed 4 times with PBS/0.05% Tween /0.02% NaN3.

A biotin-conjugated goat anti-human GROd dy was added (R&D Systems® Cat# BAF275; 100 ng/ml; 1OO ul/well). Samples were left to react for 4 h at room temperature. After washing (4 times), alkaline phosphatase-conjugated streptavidin (Jackson lmmunoresearch Cat# 016084) was added at a final dilution of 1/1000O (1OO ul/well). After 40 minutes at room temperature, plates were washed again 4 times.

P-Nitrophenyl Phosphate substrate tablets (Sigma; 5 mg Cat# N9389; 20 mg, Cat# N2765) were dissolved in diethanolamine buffer pH 9.8 to give a final tration of 1 mg/ml. 100 pl were added to each well and the CD. was read after 1 h in a Spectra Max M5 Microplate Reader (Molecular Devices) using filters of 405 and 490 nm. 3) Calculations Data are reported as Means +/- SEM. Four parameter curve fitting was used for ELISA ations. IC50 values for inhibition of IL-6 and GRO-d secretion by antibodies were calculated using leit (FIT model 205). (iv) Results 1) Assay on C20A4CI6 cells (human ocyte cell line) Both XAB4 and XAB1 are able to neutralize the induction of hulL-6 secretion by CZOA4CI6 cells ated with rhulL-17A and rhulL-17A/F in the presence of rhuTNFd.

W0 2014/122613 1 38 Control antibody (Simulect®) at 100 nM has no effect. |C5o values (means 1r SEM) for XAB4 and XAB1 are reported in Table 27. No inhibition on hulL-17F is ed even at Ab concentrations of 3 uM.

Table 27. Inhibitory effects of XAB4 and XAB1 on hulL-6 ion by C20A4C|6 cells.

Stimuli XAB4 XAB1 Control IC50 (nM) IC50 (nM) antibody (means 1r (means 1r SEM) SEM) rhulL-17A (1 nM)a 0.44 i 0.06 — rhulL-17A/F (3 nM)a 1.30 i 0.18 — rhuIL-17F (30 nM)a >3000 rhulL-17A (30 pM) + 0.024 1r 0.004 1.21 1r 0.09 rhulL-17A/F (300 pM) 0.108 1r 0.02 >10 rhulL-17F (10 nM) + >3000 >3000 >3000 a Background of hu lL-6 production without stimulation (013 1r 0.003) is cted b Background of hulL-6 production in cultures with TNF alone ( 0.20 1r 0.003) is subtracted 1O From these experiments it is evident that the parental XAB1 antibody shares neutralizing activity with its derivatives. The XAB4 variant is also seen to have a higher neutralizing activity than XAB1.

In an additional experiment, analogous to the experiment described above, all the antibodies XAB1-XAB5 were compared, as seen in Table 28. Here it can be seen that the inhibition es for XAB2, XABB and XAB5 are comparable to those ed for XAB4 and XAB1, especially to XAB4.

W0 22613 1 39 Table 28. Table Inhibitory effects of XAB antibodies on hulL-6 secretion by CZOA4C|6 cells.

Stimuli XAB1 XABZ XABB XAB4 XAB5 |C5O (nM) |C5O (nM) |C5O (nM) |C5O (nM) |C5O (nM) Means 1r Means 1r Means 1r Means 1r Means 1r rhuIL-17A O.29+0.03 O.72+0.08 063+O.15 O.51+0.04 O.55+0.01 (O.)5nM a Background of HuIL-6 production without stimuli (0.04 + 1.13 ng/ml) is subtracted 2) Assay on BJ cells (human fibroblasts) Both XAB4 and XAB1 neutralize the ion of huIL-6 and huGROd secretion by BJ cells stimulated with rhuIL-17A and 17A/F in the presence of huTNFd. Control antibody (Simulect®) at 100 nM has no effect. |C5o values for inhibition of |L-6 and hu GROd are reported in Table 29 and Table 30. Inhibition on huIL-17F is not observed even at Ab concentrations of 3 uM. From these experiments it is evident that the parental XAB1 antibody shares neutralizing activity with its derivatives.

W0 2014/122613 2014/058854 1 40 The XAB4 variant is also seen to have a higher neutralizing activity than XAB1.

Table 29. Inhibitory effect of XAB4 and XAB1 on hulL-6 secretion by BJ cells.

Stimuli XAB4 XAB1 l IC5O (nM) IC5O (nM) antibody (nM) Means 1r SEM Means 1r SEM rhuIL-17A (1er 063+0.02 rhulL-17A/F ()3nM 1.68+0.05 _ rhulL-17F (30 nM) >3ooo _ 17A 3(0 p|V|)+ 0. 012 + 0 002 0. 47 + 0. 02 >3000 rhuIL-17A/F (300)pM) 017 + 0. 01 3. 83 + 0. 63 >3000 rhuIL-17F 1(0 nIVI) >3000 >3000 >3000 a Background of hu lL-6 production without stimuli (0.32 + 0 002 ng/ml) is subtracted. b Background of hulL-6 production in cultures stimulated with TNF alone (0.45 1r 0.02 ng/ml) is subtracted.

Table 30. Inhibitory effect of XAB4 and XAB1 on -alpha secretion by BJ cells.

Stimuli XAB4 XAB1 Control IC5O (nM) IC5O (nM) antibody(nM) Means 1r SEM Means 1r SEM - < >a — lL-17F (30 nM) a >3ooo — |L-17A(30p|\/|)+ 0007+00004 072+012 |L-17A/F((300pr|) 01 +001 6.22+0.44 |L-17F 1(0 nM) >3000 >3000 >3000 W0 2014/122613 1 41 a Background of hu GROd production without stimuli (0.03 1r 0.01 ng/ml) is subtracted. b Background of hu GROd tion in cultures with TNF alone (0.15 1r 0.008 ng/ml) is subtracted.

In additional ments, analogous to the experiments described above, all the antibodies XAB1-XAB5 were compared, as seen in Table 31 and Table 32. Here it can be seen that the inhibition es for XAB2, XAB3 and XAB5 are comparable to those observed for XAB4 and XAB1, especially to XAB4.

Table 31. Inhibito effects of XAB antibodies on hulL-6 secretion b BJ cells.

Stimuli XAB1 XAB2 XAB3 XAB4 XAB5 |C50(nM) |C50(nM) lC50(nM) |C50(nM) |C50(nM) Means 1r Means 1r Means 1r Means 1r Means 1r rhuIL-17A 4.97+059 0.64+022 0.50+0002 0.55+0.04 054+002 (O.)5nM a ound of HulL-6 tion without stimuli (0.15 + 4. 06 ng/ml) is subtracted Table 32. Inhibitory effects of XAB antibodies on huGROd secretion by BJ cells.

Stimuli XAB1 XAB2 XAB3 XAB4 XAB5 |C50(nM) |C50(nM) lC50(nM) |C50(nM) |C50(nM) Means 1r Means 1r Means 1r Means 1r Means 1r rhuIL-17A 07 0.40+006 042+001 0.44+004 046+005 (O.)5nM a Background of HuGROd production without stimuli (0.03 + 0.02 ng/ml) is subtracted Example 13. In Vitro neutralization of mouse IL-17A and IL-17A/F activity by antibody variants of the disclosure CMT-93 cells (ATCC CCL-223) were cultured in RPMI (Gibco Cat# 010) supplemented with 10% fetal calf serum low lgG (Gibco Cat# 16250-078; lot 1074403), B-mercaptoethanol (5 x 10'5 M final) and Normocin (0.1 mg/ml; lnvivoGen Cat# ant-nr-2).

W0 2014/122613 1 42 The cells were detached from plastic using an Accutase on (PAA Cat# L11-007) and distributed into 96 wells microtiter plates at a y of 5x103 in 100 pl well in RPMI 1640 without fetal calf serum, B-mercaptoethanol and normocin.

The cells were allowed to adhere to the plates overnight. The next morning, rmlL-17A (SEQ ID NO: 83, MW 31000) at 1 nM, rmlL-17A/F (R&D Systems® Cat# L; MW 30400) at 3 nM, rmlL-17F (SEQ ID NO: 84; MW 30000) at 30 nM, rratlL-17A (SEQ ID NO: 85; MW 31000) at 1 nM or control medium were added in a volume of 50 ul to cate wells in the presence of 50 ul of different concentrations of test antibodies (XAB4 or XAB1), control antibodies (Simulect® 1.1% solution; C0011, 831179) or 1O l medium to reach the final volume of 200 ul/well and the final concentration of 1% fetal calf serum.

Culture supernatants were collected after an incubation of 24 h and KC production was measured by ELISA. (i) ELISA for detection of mouse KC tion ELISA microtiter plates were coated with a rat ouse KC MAb (R&D Systems® Cat# MAB453; 100 ul/well at 1 ug/ml) in 02% NaN3 and incubated overnight at 4°C. The following day, microtiter plates were blocked with 300 pl of PBS/2% BSA/0.02% NaN3 for 3 h at room temperature. Plates were then washed 4 times with PBS/0.05% Tween20/0.02% NaN3. Culture supernatants of CMT-93 cells (final dilution 1:5; 100 ul/well) were added.

To establish a titration curve, mouse KC (R&D Systems® # 453-KC, 100 ul/well) was titrated from 1 ng/ml to 0.016 ng/ml in 1:2 dilution steps. After an overnight incubation at room temperature, plates were washed 4 times with PBS/0.05% Tween 20/0.02% NaN3. A -conjugated goat anti-mouse KC antibody (R&D Systems® Cat# BAF453; 100 ul/well) at 0.1 ug/ml was added. Samples were left to react for 4 h at room temperature. After g (4 times), alkaline phosphatase-conjugated streptavidin (Jackson Immunoresearch Cat# 016084) was added at a final dilution of 1/10000 (100 ul/well). After 40 minutes at room temperature, plates were washed again 4 times.

P-Nitrophenyl Phosphate substrate tablets (Sigma; 5 mg Cat# N9389; 20 mg Cat# N2765) were dissolved in diethanolamine buffer pH 9.8 to give a final concentration of 1 W0 2014/122613 1 43 mg/ml. 100 pl culture supernatants were added to each well and the OD. was read after 1 h in a Spectra Max M5 Microplate Reader ular Devices) using filters of 405 and 490 nm. (ii) Calculations Data are reported as Means +/- SEM. Four parameter curve fitting was used for ELISA calculations. IC50 values for inhibition of KC secretion by antibodies were calculated using leitTM (FIT model 205). (iii) Results Both XAB4 and XAB1 are able to neutralize the induction of mouse KC secretion by 1O CMT-93 cells stimulated with mouse or rat IL-17A and mouse IL-17A/F. Control antibody (Simulect®) has no . IC50 values (means 1r SEM) for XAB4 and XAB1 are reported in Table 33. tion on hulL-17F is not ed even at Ab concentrations of 10 uM.

Table 33. Inhibitory effect of XAB4 and XAB1 on mouse KC secretion by GMT-93 cells.

Stimuli XAB4 IC5O (nM) XAB1 IC5O (nM) Control antibody Means 1r SEM Means + SEM n(M) mIL-17A( 1 n|\/|)a 13.8 i 0.48 539 + 29. 4 >3000 mIL-17A/F (3 10. 3 +106 >1000 >3000 mIL-17F (30 anl) >1 OOOO >1 OOOO >3000 A( 1 anl) 6. 7 + O. 84 467 + 25.1 >3000 a Background of KC production without stimuli (0. 07 + O. 001 ng/ml) is subtracted From these experiments it is evident that both the parental XAB1 antibody, as well as its derivates, has neutralizing activity. The XAB4 variant is also seen to have a higher neutralizing activity than XAB1.

In an additional experiment, analogous to the experiment described above, all the antibodies XAB1-XAB5 were compared, as seen in Table 34. Here it can be seen that the inhibition profiles for XABZ, XABB and XAB5 are comparable to those observed for XAB4 and XAB1, especially to XAB4.

Table 34 Inhibitory effects of XAB antibodies on KC secretion by GMT-93 cells.

Stimuli XAB1 XABZ XA83 XAB4 XAB5 IC5O (nM) IC5O (nM) IC5O (nM) IC5O (nM) IC5O (nM) Means 1r Means 1r Means 1r Means 1r Means 1r mIL-17A 128+14.2 209+O.96 7.0+O.29 7.8+O.78 (0.)15nM a Background of KC production without i (O. 19 + 5. 81 ng/ml) is cted Example 14. Rat antigen-induced arthritis assay (Rat AIA) Female Lewis rats (120-150 g) were sensitized intradermally on the back at two sites to 1O methylated bovine serum n (mBSA) homogenized 1:1 with complete Freund’s adjuvant on days -21 and -14 (0.1 ml ning 5mg/ml mBSA). On day 0, the rats were anaesthetized using a 5% isoflurane/air mixture and maintained using rane at 3.5% via a face mask for the intra-articular injections. The right knee received 50 ul of mg/ml mBSA in 5% glucose solution (antigen injected knee), while the left knee received 50 ul of 5% glucose on alone (vehicle injected knee). The diameters of the left and right knees were then measured using calipers immediately after the intra- articular injections and again on days 2, 4, and 7.

Treatments were administered by single subcutaneous injection on day -3. The antibody of the disclosure was injected at 0.15, 1.5, 15 and 116 mg/kg. Right knee ng was calculated as a ratio of left knee swelling, and the R/L knee swelling ratio d against time to give Area Under the Curve (AUC) graphs for control and treatment groups. The percentage inhibitions of the individual animals in each treatment group AUCs were calculated vs. the control group AUG (0% inhibition) using an Excel spreadsheet.

W0 2014/122613 1 45 Results are shown in Table 35. Dose d inhibition of right knee swelling was demonstrated for XAB4 with a calculated ED50 of 1.68 mg/kg s.c.

Table 35. Effects of single dose treatment with XAB4 on knee swelling from day 0 to day 7 in Lewis rat antigen-induced tis.

Antibody dose (mg/kg) Percentage inhibition of knee swelling AUC 116 77.01 $1.72" Data points represent the means 1r SEM of n=5 animals. * p<0.05 and ** p<0.01 ANOVA followed by Dunnett's test vs Control curve. 1O Similarly, dose related inhibition of knee swelling was demonstrated for XAB4 in a model using Wistar rats (data not shown), and in a model using mouse antigen-induced arthritis model (data not shown).

Example 15. Angiogenesis mechanistic model Chambers containing human |L-17A (between 150 and 200 ng), when placed subcutaneously in a mouse, cause new blood vessel growth around the implant. The amount of angiogenesis correlates with the weight of newly formed tissue in this area. lactic treatment with XAB4 at 0.01, 0.03, 0.1, 0.3, 1 and 3 mg/kg inhibited human |L-17 induced angiogenesis. The 5 higher doses all led to a potent and icant inhibition of tissue chamber weight. The 4 higher doses showed no dose dependency, however, the dose of 0.03 mg/kg was less efficacious than doses of 0.1 mg/kg and above.

W0 2014/122613 1 46 This study demonstrates that the potent angiogenic effect of lL-17A can be neutralized with an anti-lL-17A antibody and provides experimental ce of the iveness of XAB4 for human lL-17A in vivo.

Example 16. Experimental autoimmune encephalomyelitis (EAE) model The experimental mune alomyelitis (EAE) model is a known animal model for multiple sclerosis (reviewed e.g in Constantinescu et al., Br J Pharmacol 2011). It has been shown that tion of lL-17 reduces EAE severity in C57Bl/6 mice (Haak S et al 2009, JCI; 119:61-69).

Female C57Bl/6 mice (aged 9 weeks, Harlan, Germany) were immunized with a 50/50 1O e of recombinant rat myelin oligodendrocyte glycoprotein peptide (MOG1-125) (generated in-house) and complete Freund’s adjuvant (CFA, generated by adding 8 mg/ml Mycobacterium tuberculosis strain H37RA (Difco) to Incomplete Freund’s adjuvant (IFA, Sigma). Immunization was performed by aneous injection with 200 ug/animal of 25 at the base of tail on day 0. In addition, 200 ng/animal pertussis toxin (PT) was ed intraperitoneally on days 0 and 2.

Both therapeutic treatment effect and prophylactic treatment effect of XAB4 was tested.

Therapeutic treatment For the therapeutic treatment 16 mice were used (8 for XAB4 and 8 for control).

Treatment was initiated once the animals had a clinical score of at least 2.5 (severe hind limb weakness) for 3 days. After this, 15 mg/kg XAB4 or isotype control antibody was injected subcutaneously each week with a single dose.

The results are shown in Figures 11 to 15 (d.p.i is days post immunization). In all figures, XAB4 is represented by circles and the e control is represented by squares. The therapeutic score (mean+SEM) is shown in Figure 11. It is clearly seen that animals treated with XAB4 has a lower mean clinical score than isotype control.

Figure 12 shows the weight change (%) for the two groups of mice, and Figure 13 shows the cumulative therapeutic scores. Figures 14 and 15 are comparisons of the therapeutic score pre- and post-treatment. It is clearly seen in all graphs that XAB4 has W0 22613 1 47 a therapeutic effect ed to the isotype control. Thus, therapeutic ent with XAB4 significantly reduced the severity of EAE.

Prophylactic treatment For the prophylactic treatment 19 mice were used (10 for XAB4 and 9 for control). Each animal was treated one day prior to immunization with 15 mg/kg XAB4 or isotype control, through a single subcutaneous injection. After this, 15 mg/kg XAB4 or isotype control antibody was injected subcutaneously each week with a single dose.

The results are shown in Figures 16 to 20 (d.p.i is days post immunization). ln Figures 16 to 19, XAB4 is represented by closed circles and the isotype control is represented 1O by open squares. The prophylactic score (mean+SEM) is shown in Figure 16. It is y seen that animals treated with XAB4 has a lower mean clinical score. Figure 17 shows the weight change (%) for the two groups of mice, and Figure 18 shows the cumulative prophylactic scores. The m prophylactic score is seen in Figure 19. It is clearly seen in all graphs that XAB4 has an effect compared to the isotype control.

Furthermore, in Figure 20, where XAB4 is represented by a solid line and isotype control is represented by a dotted line, it is seen that EAE onset is later for the group of mice treated with XAB4, compared to the group of mice treated with isotype control.

Thus, it is shown that prophylactic treatment with XAB4 significantly delayed EAE onset and reduced maximal EAE severity.

Example 17. Attenuation of IL17A-induced levels of IL6, CXCL1, IL-8, GM-CSF, and CCL2 in human astrocytes The effects of XAB4 on the levels of lL-6, CXCL1, lL-8, GM-CSF, and CCL2 in astrocytes isolated from the cerebral cortex of the human brain were investigated.

Astrocytes release a number of growth factors, cytokines and chemokines that allow them to regulate cellular communication, migration and survival of al, glial and immune cells. The direct ication of astrocytic et with endothelial cells also allows astrocytes to control function of the blood-brain-barrier. Moreover, astrocytes release and uptake ransmitters, such as glutamate, at the synaptic cleft that allow them to regulate synaptic transmission and excitoxicity. It is significant that astrocytes form scar pathology after CNS injury, thus having apparent opposing roles in normal W0 2014/122613 2014/058854 1 48 physiology and hysiology. In e, astrocytes are suggested to play roles in a range of psychiatric, neurological and neurodegenerative disorders, where their role in neuroinflammation is likely to be important.

The data showed that co-stimulation with lL-17A and TNFd ed the release of IL- 6, CXCL1, lL-8, GM-CSF, and CCL2, and that XAB4 inhibited levels of lL-6, CXCL1, IL- 8, GM-CSF, and CCL2 in human astrocytes. These data indicate a dominate role for lL- 17A in cytokine release from astrocytes and support their use as drug targets for neuroinflammatory diseases. It is noteworthy that the pretreatment of human astrocytes with XAB4 inhibited lL-17A-induced and lL-17A/TNFd-induced, without affecting TNFd- 1O induced, levels of lL-6, CXCL1, lL-8, GM-CSF, and CCL2. Taken together, the data suggested that selective inhibition of lL-17A signaling with XAB4 ates the level of pro-inflammatory cytokines in human astrocytes. ln disease, ytes are suggested to play roles in a range of psychiatric, neurological and neurodegenerative disorders, where their role in neuroinflammation is likely to be important. Novel drugs that alter astrocyte function are thus of potential value, where regulation of yte function may prove therapeutically useful. Consequently, since XAB4 was shown to have an effect on lL-6, CXCL1, lL-8, GM-CSF, and CCL2 production of astrocytes, it can be concluded that XAB4 may be a useful therapeutic agent, such as for treatment of Multiple Sclerosis (MS).

Materials and Methods All cytokines were sed from R&D s. Basiliximab (Novartis, Basel, Switzerland) was used as isotype control. Primary antibodies used were: anti-lL17RA Alexa Fluor 647 (BG/hlL17AR, Biolegend), anti-lL17RC Alexa Fluor 488 (309822, R&D Systems, UK), 65 (Santa Cruz, USA), mouse lgG Alexa Fluor 647 (MOPC-21, Biolegend, UK), mouse lgG Alexa Fluor 488 (133303, R&D , UK), mouse lgG Biotin (G155-178, BD Biosciences, Switzerland) and rat lgG PE (A95-1, BD Biosciences, rland). Secondary antibodies and dyes used were: biotinylated goat anti-rabbit lgG (BA1000, Vector, UK), streptavidin conjugated Alexa Fluor 488 and Alexa Fluor 633 (S11223 and S2137, Life Technology, USA), goat anti-mouse Alexa Fluor 488 and Alexa Fluor 633 (A1101 and A21050, Life Technology, USA), streptavidin BV421 (405226, Biolegend, UK), Hoechst 34580 (H21486, Life Technology, USA).

W0 2014/122613 1 49 Human ytes d from cerebral cortex were purchased from ScienCell Research Laboratory (USA) ogue number 1800). Cells were grown as per provider’s instructions. Briefly cells were grown in human astrocyte media (ScienCell catalogue number 1801) supplemented with 1% astrocyte growth supplement (ScienCell catalogue number 1852), 5% fetal calf serum (ScienCell catalogue number 0010) and 1% Penicillin/Streptomycin (ScienCell catalogue number 0503). Cells were maintained in T75 e flasks at 5% C02 and 37°C with the media changed every three days until 80% confluent. For all treatments, 70,000 cells well plated in 24-well plates, grown for 3 days, serum starved for 2-4 hr, after which astrocytes were treated 1O for 2 hr with XAB4, and fter treated for 18-20 hr with recombinant human cytokines as indicated in the figure legends. The cell pellets were used to quantify mRNA levels of cytokines by qPCR and the supernatants were used to quantify the protein levels of cytokines by HTRF o, France, used for lL-6, lL-8 & CXCL1) or AlphaLlSA (PerkinElmer, USA, used for CCL2 & GM-CSF).

Measurement of cytokine mRNA was performed by real time-polymerase chain on (RT-PCR). Briefly, ytes were lysed for 5 min at room temperature by gently shaking in 350 pl lysis buffer (RLT buffer with 1% B-mercaptoethanol) and total RNA was extracted using RNeasy Microkit (74004, , Switzerland). The cDNA was synthesized using SuperScript lll reverse transcriptase (18080-400, Life Technology, Switzerland). The expression level of each gene was assessed by q-PCR in a Viia7 Real-time PCR e (Life logy, Switzerland). Taqman probes were purchased from Life Technology, Switzerland. Each sample was analyzed in triplicate and normalized to hypoxanthine-guanine phosphoribosyltransferase (HPRT). Levels of human IL6, lL8, CXCL1 protein (ng/ml) in human astrocyte supernatant (10 ul) were assessed by HTRF (IL6: 62lL6PEC; lL8: 62lL8PEC; CXCL1: 6FGROPEG, Cisbio, France) and the level of human CCL2 protein (ng/ml) in human astrocyte supernatant (5 ul) was assessed by lSA human CCL2/MCP1 (AL244C, PerkinElmer, USA). All measurements were performed according to manufacturer’s instructions.

Cells suspensions of human astrocytes were obtained from adherent cultures using PBS-5mM EDTA. For extracellular staining cells were incubated with whole mouse lgG for 10 min at 4° C in PBS 2% BSA, and then d with antibodies for 30 min at 4° C in PBS 2% BSA. For intracellular staining, cells were permeabilized with W0 2014/122613 1 50 Cytofix/Cytoperm solution (554714, BD Biosciences, Switzerland) for 20 min at 4°C before incubating with antibodies for 30 min at 4°C. After filtration through 70 um strainer, cells were acquired on a BDFortessa (BD Biosciences, Switzerland) and data analyzed using FlowJo software (Tree Star Inc, USA).

After compound treatment, cells were washed in PBS (Sigma Aldrich, Germany) followed by on in ice-cold 100% methanol for 10 min. Cells were washed 3 x 5 min in sterile PBS then permeabilized by incubation with 0.2% Triton-X-100 (Sigma Aldrich, Germany) in PBS for 5 min at room temperature. Non-reactive sites were blocked overnight at +4°C with blocking buffer which consisted of 10% normal goat serum (Life 1O Technology, USA) and 2% bovine serum albumin (Sigma h, Germany) in PBS.

The cells were then incubated in primary antibody overnight at 4°C. The primary antibody was removed and the cells washed 3 x 5 min PBS after which the secondary fluorescent dy was applied for 2 hr at room ature. The coverslips were then washed 5 x 5 min in PBS and counter stained with Hoescht 34580 nuclear stain. The coverslips were finally mounted on cope slides in VectashieldR mounting medium (Vector, UK) and the edges of the lip sealed with nail varnish. The cells were imaged using a Zeiss LSM 510 META confocal laser scanning microscope utilizing an Axiovert 200M inverted microscope (Zeiss Ltd, Germany).

Results Antagonism of TNF-d or lL-17A stimulation, or lL-17A/TNF-d co-stimulation by XAB4 is shown in Figures 21 A to 25 A. Antagonism of lL-1B or lL-17A/ lL-1B co-stimulation by XAB4 is shown in Figures 21 B to 25 B.

Figure 21 shows antagonistic effect on lL-6 e, Figure 22 shows antagonistic effect on CXCL1 release, Figure 23 shows antagonistic effect on lL-8, Figure 24 shows antagonistic effect on GM-CSF and Figure 25 shows antagonistic effect on CCL2.

Primary human ytes were treated with increasing concentrations of XAB4 (0.01nM, 0.1nM 1 nM and 10nM), with or without lL-17A (50ng/ml), TNF-d (10 , lL-1B, lL-17A/TNF-d and lL-1B/TNF-o. All trations used are indicated in the figures. The data shown is a representative of two experiments for XAB4 0.01nM, and W0 2014/122613 1 51 of three ments for XAB4 0.1nM, 1nM and 10nM. Values shown are means 1r S.E.M.

As seen in Figure 21A, XAB4 (all concentrations) has an antagonistic effect, ed to both control and isotype, on release of lL-6 from astrocytes stimulated with lL-17A, or lL-17A/TNF-d. Concentration of lL-6 (ng/ml) is represented by the y-axis and concentration of XAB4 is represented on the x-axis (0, Le. control, 0.01nM, 0.1nM 1 nM and 10nM) for each dataset, and 10 nM for isotype. The dataset to the left relates to unstimulated cells, the next dataset relates to cells stimulated with TNF-d (10 ng/ml), the next dataset relates to cells stimulated with lL-17A (50ng/ml) and the last dataset 1O relates to cells co-stimulated with TNF-d (10 ng/ml) and lL-17A (50ng/ml). The last dataset has about 10 fold higher scale of the y-axis. As seen in Figure 218, XAB4 (all trations) has no antagonistic effect on cells stimulated with lL-1B (0.1 ng/ml) or co-stimulated with lL-1B (0.1 ng/ml) and lL-17A (50 ng/ml), compared to isotype.

As seen in Figure 22A, XAB4 (all concentrations) has an antagonistic effect, compared to both control and e, on release of CXCL1 from astrocytes stimulated with lL- 17A, or lL-17A/TNF-d. Concentration of CXCL1 ) is represented by the y-axis and concentration of XAB4 is ented on the x-axis (0, Le. control, 0.01nM, 0.1nM 1 nM and 10nM) for each dataset, and 10 nM for isotype. The dataset to the left relates to unstimulated cells, the next dataset s to cells ated with TNF-d (10 ng/ml), the next dataset relates to cells stimulated with lL-17A (50ng/ml) and the last dataset relates to cells co-stimulated with TNF-d (10 ng/ml) and lL-17A (50ng/ml). The last dataset has about 10 fold higher scale of the y-axis. As seen in Figure 228, XAB4 (all concentrations) has no antagonistic effect on cells stimulated with lL-1B (0.1 ng/ml) or co-stimulated with lL-1B (0.1 ng/ml) and lL-17A (50 ng/ml), compared to e.

As seen in Figure 23A, XAB4 (all concentrations) has an antagonistic effect, compared to control, on release of lL-8 from astrocytes stimulated with , or lL-17A/TNF-d.

Compared to isotype, XAB4 (0.1nM, 1nM and 10nM) has an antagonistic effect on release of lL-8. Concentration of lL-8 (ng/ml) is represented by the y-axis and concentration of XAB4 is represented on the x-axis (0, Le. control, 0.01nM, 0.1nM 1 nM and 10nM) for each t, and 10 nM for isotype. The dataset to the left s to unstimulated cells, the next dataset relates to cells stimulated with TNF-d (10 ng/ml), the next dataset relates to cells stimulated with lL-17A (50ng/ml) and the last dataset W0 2014/122613 1 52 relates to cells mulated with TNF-d (10 ng/ml) and lL-17A ml). The last dataset has about 5 fold higher scale of the y-axis. As seen in Figure 238, XAB4 (all concentrations) has no antagonistic effect on cells stimulated with lL-1B (0.1 ng/ml) or co-stimulated with lL-1B (0.1 ng/ml) and lL-17A (50 ng/ml), compared to isotype.

As seen in Figure 24A, XAB4 (all concentrations) has an antagonistic effect, compared to both control and isotype, on release of GM-CSF from astrocytes stimulated with lL- 17A/TNF-d. XAB4 (0.1nM, 1nM and 10nM) has an antagonistic effect on e of GM- CSF from astrocytes stimulated with lL-17A, compared to isotype and control.

Concentration of GM-CSF (ng/ml) is represented by the y-axis and concentration of 1O XAB4 is ented on the x-axis (0, Le. l, 0.01nM, 0.1nM 1 nM and 10nM) for each dataset, and 10 nM for isotype. The dataset to the left s to unstimulated cells, the next dataset relates to cells stimulated with TNF-d (10 , the next dataset relates to cells stimulated with lL-17A (50ng/ml) and the last dataset relates to cells co- stimulated with TNF-d (10 ng/ml) and lL-17A (50ng/ml). As seen in Figure 248, XAB4 (all concentrations) has no or low antagonistic effect on cells stimulated with lL-1B (0.1 ng/ml) or co-stimulated with lL-1B (0.1 ng/ml) and lL-17A (50 ng/ml), compared to isotype.

As seen in Figure 25A, XAB4 (all concentrations) has an antagonistic effect, compared to both control and isotype, on release of CCL2 from astrocytes stimulated with lL-17A.

XAB4 (0.1nM, 1nM and 10nM) has an antagonistic effect on release of CCL2 from astrocytes stimulated with lL-17A/TNF-d, compared to isotype and control.

Concentration of CCL2 (ng/ml) is represented by the y-axis and concentration of XAB4 is represented on the x-axis (0, Le. control, 0.01nM, 0.1nM 1 nM and 10nM) for each dataset, and 10 nM for isotype. The dataset to the left relates to ulated cells, the next t relates to cells stimulated with TNF-o (10 ng/ml), the next dataset relates to cells stimulated with lL-17A (50ng/ml) and the last t relates to cells co-stimulated with TNF-d (10 ng/ml) and lL-17A (50ng/ml). As seen in Figure 258, XAB4 (all concentrations) has no antagonistic effect on cells stimulated with lL-1B (0.1 ng/ml) or co-stimulated with lL-1B (0.1 ng/ml) and lL-17A (50 ng/ml), compared to isotype.

Taken together, the data suggested that ive inhibition of lL-17A ing with XAB4 attenuates the level of pro-inflammatory cytokines in human astrocytes. ln disease, astrocytes are suggested to play roles in a range of psychiatric, neurological W0 2014/122613 1 53 and neurodegenerative disorders, where their role in neuroinflammation is likely to be important. Since XAB4 was shown to have an effect on lL-6, CXCL1, lL-8, GM-CSF, and CCL2 production of astrocytes, XAB4 may be a useful therapeutic agent, such as for ent of Multiple Sclerosis (MS).

Sequence ation Sequence data relating to XABl, XABZ, XABS, XAB4 and XAB5 is ized below for ease of reference.

Table 1 describes the amino acid sequences (SEQ ID NOs) of the full length heavy and light chains of examples XABl, XABZ, XABS, XAB4 and XAB5. 1O The antibodies XABl, XABZ, XABS, XAB4 or XAB5 can be produced using conventional antibody inant production and purification processes. For example, the coding sequences as described in Table 3 or Table 4 are cloned into a production vector for recombinant expression in mammalian production cell line.

Table 2 summarizes the le heavy (VH) and light chain (VL) amino acid sequence of XABl, XABZ, XABS, XAB4 or XAB5, which can be used to generate chimeric antibodies from XABl, XABZ, XABS, XAB4 or XAB5.

Table 5 summarizes the useful CDR sequences of XABl, XABZ, XABS, XAB4 and XAB5 (plus consensus sequences) to generate alternative CDR grafted antibodies, wherein the CDR regions from XABl, XABZ, XABS, XAB4 and XAB5 are defined according to Kabat definition.

Table 6 summarizes the useful CDR ces of XABl, XABZ, XABS, XAB4 and XAB5 (plus consensus sequences) to generate alternative CDR grafted antibodies, wherein the CDR regions from XABl, XABZ, XABS, XAB4 and XAB5 are defined ing to Chothia definition.

All the sequences referred to in this specification (SEQ ID NOs) are found in Table 36.

W0 2014/122613 1 54 SEQUENCE LIST Useful amino acids and nucleotide sequences for practicing the invention are found in Table 36.

Table 36. Sequence list Antibody/ Sequence Amino acid sequence or cleotide sequence Fragment Identifier (PN) (SEQ ID NO:) CHOTHIA (CHOTHIA) CHOTHIA (CHOTHIA) —-_(CHOTHIA) —_(CHOTHIA) —_(KABAT) (KABAT) —_(KABAT) (KABAT) —_—KABAT SGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSS XAB1, VL AIQLTQSPSSLSASVGDRVTITCRPSQGIISALAVWQ QKPGKAPKLLIYDASSLENGVPSRFSGSGSGTDFTL TISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK XAB‘I, HEAVY CHAIN EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLM VTCVVVDVSHEDPEVKFNVWVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE 1 55 YKCKVSNKALPAMEKHSKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRVWQQGNVFSC LHNHYTQKSLSLSPGK XAB‘I, LIGHT CHAIN AIQLTQSPSSLSASVGDRVTITCRPSQGI ISALAVWQ QKPGKAPKLUYDASSLENGVPSRFSGSGSGTDFTL EDFATYYCQQFNSYPLTFGGGTKVHKRTV AAPSVHFPPSDEQLKSGTASVVCLLNNFYPREAKV CMNKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC PN ENCODING SEQ GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 12 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGDMNACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA GGACACCGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACUUTGGGGCCAGGGCACCCT CGTGTCCAGC PN ENCODING SEQ GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT ID NO: 13 CTGCATCTGTGGGAGACAGAGTCACCATCACTTG CCGGCCAAGTCAGGGCATTATCAGTGCTTTAGCC TGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGC TCCTGATCTATGATGCCTCCAGTTTGGAAAATGGG GTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGA CAGATTTCACTCTCACCATCAGCAGCCTGCAGCC TGAAGATTTTGCAACTTATTACTGTCAACAGTTTAA TAGTTACCCTCTCACTTTCGGCGGAGGGACCAAG GTGGAGATCAAA PN ENCODING SEQ GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 14 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGDMNACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA GGACACCGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACUUTGGGGCCAGGGCACCCT GGTCACCGTGTCCAGCGCTAGCACCAAGGGCCC CAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAG CACCAGCGGCGGCACAGCCGCCCTGGGCTGCCT GGACTACTTCCCCGAGCCCGTGACCGT GAACAGCGGAGCCCTGACCTCCGGCGT GCACACCTTCCCCGCCGTGCTGCAGAGCAGCGG CAGCCTGTCCAGCGTGGTGACAGTGCCC AGCAGCAGCCTGGGCACCCAGACCTACATCTGCA ACGTGAACCACAAGCCCAGCAACACCAAGGTGGA CAAGAGAGTGGAGCCCAAGAGCTGCGACAAGAC CCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCC AAGCCCAAGGACACCCTGATGATCAGCAGGACCC CCGAGGTGACCTGCGTGGTGGTGGACGTGAGCC 2014/058854 ACGAGGACCCAGAGGTGAAGTTCAACTGGTACGT GGACGGCGTGGAGGTGCACAACGCCAAGACCAA GCCCAGAGAGGAGCAGTACAACAGCACCTACAG GGTGGTGTCCGTGCTGACCGTGCTGCACCAGGA CTGGCTGAACGGCAAGGAATACAAGTGCAAGGTC TCCAACAAGGCCCTGCCAGCCCCCATCGAAAAGA CCATCAGCAAGGCCAAGGGCCAGCCACGGGAGC CCCAGGTGTACACCCTGCCCCCCTCCCGGGAGG AGATGACCAAGAACCAGGTGTCCCTGACCTGTCT GGTGAAGGGCTTCTACCCCAGCGACATCGCCGT GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAA CTACAAGACCACCCCCCCAGTGCTGGACAGCGAC GGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGG ACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCA GCTGCAGCGTGATGCACGAGGCCCTGCACAACC ACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGG CAAG PN ENCODING SEQ GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT |E)NO:15 CTGCATCTGTGGGAGACAGAGTCACCATCACTTG CCGGCCAAGTCAGGGCATTATCAGTGCTTTAGCC TGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGC TCCTGATCTATGATGCCTCCAGTTTGGAAAATGGG GTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGA CAGATTTCACTCTCACCATCAGCAGCCTGCAGCC TTTTGCAACTTATTACTGTCAACAGTTTAA TAGTTACCCTCTCACTTTCGGCGGAGGGACCAAG GTGGAGATCAAACGTACGGTGGCCGCTCCCAGC GTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGA AGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGA ACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTG GGACAACGCCCTGCAGAGCGGCAACAG GAGCGTCACCGAGCAGGACAGCAAGGA CTCCACCTACAGCCTGAGCAGCACCCTGACCCTG GCCGACTACGAGAAGCATAAGGTGTACG CCTGCGAGGTGACCCACCAGGGCCTGTCCAGCC CCGTGACCAAGAGCTTCAACAGGGGCGAGTGC XAB2, CDRH1 GFTFSSY (CHOTHIA) XAB2, CDRH2 KQDGSE (CHOTHIA) XAB2, CDRH3 DRGSLYY (CHOTHIA) XAB2, CDRL1 SQVHSA (CHOTHIA) XAB2, CDRL2 DAS (CHOTHIA) XAB2, CDRL3 FDSYPL CHOTHIA XAB2, CDRH1 SYWWE (KABAT) XAB2, CDRH2 MKQDGSEKYYVDSVKG KABAT XAB2, CDRH3 DRGSLYY (KABAT) KABAT (KABAT) —-—KABAT EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSS XABZ VL SPSSLSASVGDRVTITCRPSQVI ISALAVWQ QKPGKAPKLLIYDASSLEQGVPSRFSGSVSGTDFTL TISSLQPEDFATYYCQQFDSYPLTFGGGTKVEIK XABZ, HEAVY CHAIN :- EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLM ISRTPEVTCVVVDVSHEDPEVKFNVWVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK XABZ, LIGHT CHAIN AIQLTQSPSSLSASVGDRVTITCRPSQVI ISALAVWQ QKPGKAPKLLIYDASSLEQGVPSRFSGSVSGTDFTL TISSLQPEDFATYYCQQFDSYPLTFGGGTKVEI KRTV AAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC PN ENCODING SEQ 16 GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 12 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC GGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGTACTACGTGGACAGCGTGAAGGGCC CCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA GGACACCGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACTATTGGGGCCAGGGCACCCT GGTCACCGTGTCCAGC PN ENCODING SEQ CAGCTGACCCAGAGCCCCAGCAGCCTG ID NO: 25 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC CCCAGCCAGGTCATCATCAGCGCCCTG GCCTGGTATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAC AGGGCGTGCCCAGCCGGTTCAGCGGCAGCGTGT CCGGCACCGACTTCACCCTGACCATCAGCTCCCT GCAGCCCGAGGACTTCGCCACCTACTACTGCCAG CAGTTCGACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAG PN ENCODING SEQ GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG WO 22613 IDNOVM GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC AGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGDMNACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA GGACACCGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACUUTGGGGCCAGGGCACCCT GGTCACCGTGTCCAGCGCTAGCACCAAGGGCCC GTTCCCCCTGGCCCCCAGCAGCAAGAG CACCAGCGGCGGCACAGCCGCCCTGGGCTGCCT GGTGAAGGACTACTTCCCCGAGCCCGTGACCGT GTCCTGGAACAGCGGAGCCCTGACCTCCGGCGT GCACACCTTCCCCGCCGTGCTGCAGAGCAGCGG CCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCC AGCCTGGGCACCCAGACCTACATCTGCA ACGTGAACCACAAGCCCAGCAACACCAAGGTGGA CAAGAGAGTGGAGCCCAAGAGCTGCGACAAGAC CCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCC AAGCCCAAGGACACCCTGATGATCAGCAGGACCC CCGAGGTGACCTGCGTGGTGGTGGACGTGAGCC ACGAGGACCCAGAGGTGAAGTTCAACTGGTACGT GGACGGCGTGGAGGTGCACAACGCCAAGACCAA GCCCAGAGAGGAGCAGTACAACAGCACCTACAG GGTGGTGTCCGTGCTGACCGTGCTGCACCAGGA CTGGCTGAACGGCAAGGAATACAAGTGCAAGGTC TCCAACAAGGCCCTGCCAGCCCCCATCGAAAAGA CCATCAGCAAGGCCAAGGGCCAGCCACGGGAGC CCCAGGTGTACACCCTGCCCCCCTCCCGGGAGG CCAAGAACCAGGTGTCCCTGACCTGTCT GGTGAAGGGCTTCTACCCCAGCGACATCGCCGT GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAA CTACAAGACCACCCCCCCAGTGCTGGACAGCGAC GGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGG ACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCA GCTGCAGCGTGATGCACGAGGCCCTGCACAACC ACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGG CAAG PN ENCODING SEQ GCCATCCAGCTGACCCAGAGCCCCAGCAGCCTG |DNCt26 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC TGTCGGCCCAGCCAGGTCAHJUCAGCGCCCTG GCCTGGTATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAC AGGGCGTGCCCAGCCGGTTCAGCGGCAGCGTGT CCGGCACCGACTTCACCCTGACCATCAGCTCCCT GCAGCCCGAGGACTTCGCCACCTACTACTGCCAG CAGTTCGACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAGCGTACGGTGGCCG CTCCCAGCGTGTTCAKNTCCCCCCCAGCGACGA GCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTG CCTGCTGAACAACTTCTACCCCCGGGAGGCCAAG GTGCAGTGGAAGGTGGACAACGCCCTGCAGAGC GGCAACAGCCAGGAGAGCGTCACCGAGCAGGAC GACTCCACCTACAGCCTGAGCAGCACCC TGACCCTGAGCAAGGCCGACTACGAGAAGCATAA GGTGTACGCCTGCGAGGTGACCCACCAGGGCCT GTCCAGCCCCGTGACCAAGAGCTTCAACAGGGG ALTERNATIVE PN GAGGTGCAGCTGGTGGAATCAGGAGGCGACCTG NG SEQ ID GTGCAGCCTGGCGGCTCACTGAGACTGAGCTGC NOVQ GCCGCTAGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC TGGAGTGGGTGGCCAAUUTAAGCAGGACGGCTC AGAGAAGTACTACGTGGACTCAGTGAAGGGCCG GTTCACTATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCT ATIVE PN GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGA ENCODING SEQ ID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NO:25 TAGACCTAGTCAGGTGATCATTAGCGCCCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG CTGCTGATCTACGACGCTAGTAGTCTGGAACAGG GCGTGCCCTCTAGGTTTNSCGGCTCAGTGTCAGG CACCGACTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TCGATAGCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAG ALTERNATIVE PN GAGGTGCAGCTGGTGGAATCAGGAGGCGACCTG ENCODING SEQ ID GTGCAGCCTGGCGGCTCACTGAGACTGAGCTGC NO:14 GCCGCTAGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC TGGAGTGGGTGGCCAAUUTAAGCAGGACGGCTC AGAGAAGTACTACGTGGACTCAGTGAAGGGCCG GTTCACTATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCTGCTAGCACCAAGGGCCCAAGTG TCTTTCCCCTGGCCCCCAGCAGCAAGTCCACAAG CGGAGGCACTGCAGCTCTGGGTTGTCTGGTGAA GGACTACTTCCCCGAGCCCGTGACAGTGTCCTGG AACAGCGGAGCCCTGACCTCCGGCGTGCACACC TTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTAC AGCCTGAGCAGCGTCGTGACTGTGCCTAGTTCCA GCCTGGGCACCCAGACCTATATCTGCAACGTGAA CCACAAGCCCAGCAACACCAAGGTGGACAAGAGA GTGGAGCCCAAGAGCTGCGACAAGACCCACACC CCCTGCCCAGCTCCAGAACTGCTGGGA GGACCCAGCGTGTTCCTGTTCCCCCCCAAGCCCA AGGACACCCTGATGATCAGCAGGACCCCCGAGG GCGTGGTGGTGGACGTGTCCCACGAGG ACCCAGAGGTGAAGTTCAACTGGTACGTGGACGG GGTGGAGGTGCACAACGCCAAGACCAAGCCCAG AGAGGAGCAGTACAACAGCACCTACAGGGTGGT GTCCGTCCTGACAGTGCTGCACCAGGATTGGCTG AACGGCAAAGAATACAAGTGCAAAGTCTCCAACA W0 22613 AGGCCCTGCCAGCCCCAATCGAAAAGACAATCAG CAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGT GTACACCCTGCCCCCCAGCCGGGAGGAGATGAC CAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAG GGCTTCTACCCCAGCGATATCGCCGTGGAGTGG AACGGCCAGCCCGAGAACAACTACAAG ACCACCCCCCCAGTGCTGGACAGCGACGGCAGC TTCTTCCTGTACAGCAAGCTGACCGTGGACAAGT CCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCA GCGTGATGCACGAGGCCCTGCACAACCACTACAC CCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG ALTERNAHVEPN CAGCTGACTCAGTCACCTAGTAGCCTGA KSSEQID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NCX26 TAGACCTAGTCAGGTGATCATTAGCGCCCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG CTGCTGATCTACGACGCTAGTAGTCTGGAACAGG GCGTGCCCTCTAGGTTTNSCGGCTCAGTGTCAGG CACCGACTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TCGATAGCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCC TTCKKNTCCCCCCCAGCGACGAGCAGC TGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGC TGAACAACTTCTACCCCCGGGAGGCCAAGGTGCA GTGGAAGGTGGACAACGCCCTGCAGAGCGGCAA CAGCCAGGAGAGCGTCACCGAGCAGGACAGCAA GGACTCCACCTACAGCCTGAGCAGCACCCTGACC CTGAGCAAGGCCGACTACGAGAAGCATAAGGTGT ACGCCTGCGAGGTGACCCACCAGGGCCTGTCCA GCCCCGTGACCAAGAGCTTCAACAGGGGCGAGT XAB3, CDRH1 GFTFSSY (CHOTHIA) XAB3, CDRH2 N KQDGSE CHOTHIA XAB3, CDRH3 00 DRGSLYY (CHOTHIA) XAB3, CDRL1 SQGWWE (CHOTHIA) XAB3, CDRL2 O1 DAS (CHOTHIA) XAB3, CDRL3 FNSYPL (CHOTHIA) XAB3, CDRH1 SYWWB (KABAT) XAB3, CDRH2 MKQDGSEKYYVDSVKG (KABAT) XAB3, CDRH3 00 DRGSLYY KABAT XAB3, CDRL1 RPSQGIYWELA (KABAT) XAB3, CDRL2 DASSLEQ KABAT XAB3, CDRL3 _\ QQFNSYPLT 1 61 ) 2—— EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSS XAB3 VL - AIQLTQSPSSLSASVGDRVTITCRPSQGIYWELAVW QQKPGKAPKLLIYDASSLEQGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK XAB3, HEAVY CHAIN EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLM ISRTPEVTCVVVDVSHEDPEVKFNVWVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE NKALPAPI EKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK XAB3, LIGHT CHAIN AIQLTQSPSSLSASVGDRVTITCRPSQGIYWELAVW QQKPGKAPKLLIYDASSLEQGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRT VAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC PN ENCODING SEQ GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 12 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC AGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGTACTACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA GGACACCGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACTATTGGGGCCAGGGCACCCT CGTGTCCAGC PN ENCODING SEQ GCCATCCAGCTGACCCAGAGCCCCAGCAGCCTG ID NO: 35 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC TGTCGGCCCAGCCAGGGCATCTACTGGGAGCTG GCCTGGTATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAC AGGGCGTGCCCAGCCGGTTCAGCGGCAGCGGAT CCGGCACCGACTTCACCCTGACCATCAGCTCCCT CGAGGACTTCGCCACCTACTACTGCCAG CAGTTCAACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAG PN ENCODING SEQ GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 14 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC GGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGTACTACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA CGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACUUTGGGGCCAGGGCACCCT CGTGTCCAGCGCTAGCACCAAGGGCCC CAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAG CACCAGCGGCGGCACAGCCGCCCTGGGCTGCCT GGTGAAGGACTACTTCCCCGAGCCCGTGACCGT GTCCTGGAACAGCGGAGCCCTGACCTCCGGCGT GCACACCTTCCCCGCCGTGCTGCAGAGCAGCGG CCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCC AGCAGCAGCCTGGGCACCCAGACCTACATCTGCA ACGTGAACCACAAGCCCAGCAACACCAAGGTGGA AGTGGAGCCCAAGAGCTGCGACAAGAC CCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCC AAGCCCAAGGACACCCTGATGATCAGCAGGACCC CCGAGGTGACCTGCGTGGTGGTGGACGTGAGCC ACGAGGACCCAGAGGTGAAGTTCAACTGGTACGT CGTGGAGGTGCACAACGCCAAGACCAA GCCCAGAGAGGAGCAGTACAACAGCACCTACAG GGTGGTGTCCGTGCTGACCGTGCTGCACCAGGA CTGGCTGAACGGCAAGGAATACAAGTGCAAGGTC AAGGCCCTGCCAGCCCCCATCGAAAAGA CCATCAGCAAGGCCAAGGGCCAGCCACGGGAGC CCCAGGTGTACACCCTGCCCCCCTCCCGGGAGG AGATGACCAAGAACCAGGTGTCCCTGACCTGTCT GGTGAAGGGCTTCTACCCCAGCGACATCGCCGT GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAA CTACAAGACCACCCCCCCAGTGCTGGACAGCGAC GGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGG ACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCA GCTGCAGCGTGATGCACGAGGCCCTGCACAACC ACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGG CAAG PN ENCODING SEQ GCCATCCAGCTGACCCAGAGCCCCAGCAGCCTG |DNCt36 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC TGTCGGCCCAGCCAGGGCAKNACTGGGAGCTG GCCTGGTATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAC AGGGCGTGCCCAGCCGGTTCAGCGGCAGCGGAT CCGGCACCGACTTCACCCTGACCATCAGCTCCCT GCAGCCCGAGGACTTCGCCACCTACTACTGCCAG CAGTTCAACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAGCGTACGGTGGCCG CTCCCAGCGTGTTCKKNTCCCCCCCAGCGACGA GCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTG CCTGCTGAACAACTTCTACCCCCGGGAGGCCAAG GTGCAGTGGAAGGTGGACAACGCCCTGCAGAGC GGCAACAGCCAGGAGAGCGTCACCGAGCAGGAC AGCAAGGACTCCACCTACAGCCTGAGCAGCACCC TGACCCTGAGCAAGGCCGACTACGAGAAGCATAA GGTGTACGCCTGCGAGGTGACCCACCAGGGCCT GTCCAGCCCCGTGACCAAGAGCTTCAACAGGGG CGAGTGC ALTERNATIVE PN GAGGTGCAGCTGGTGGAATCAGGAGGCGACCTG ENCODING SEQ ID CCTGGCGGCTCACTGAGACTGAGCTGC NOVQ AGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC TGGAGTGGGTGGCCAAUUTAAGCAGGACGGCTC AGAGAAGTACTACGTGGACTCAGTGAAGGGCCG GTTCACTATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCT ALTERNATIVE PN GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGA ENCODING SEQ ID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NCI35 TAGACCTAGCCAGGGAATCTACTGGGAGCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG CTGCTGATCTACGACGCTAGTAGTCTGGAACAGG GCGTGCCCTCTAGGTTTNSCGGCTCAGGCTCAGG CTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TTAACTCCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAG ALTERNATIVE PN GAGGTGCAGCTGGTGGAATCAGGAGGCGACCTG ENCODING SEQ ID GTGCAGCCTGGCGGCTCACTGAGACTGAGCTGC NO:14 GCCGCTAGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC TGGAGTGGGTGGCCAAUUTAAGCAGGACGGCTC AGAGAAGTACTACGTGGACTCAGTGAAGGGCCG GTTCACTATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCTGCTAGCACCAAGGGCCCAAGTG TCTTTCCCCTGGCCCCCAGCAGCAAGTCCACAAG CGGAGGCACTGCAGCTCTGGGTTGTCTGGTGAA GGACTACTTCCCCGAGCCCGTGACAGTGTCCTGG AACAGCGGAGCCCTGACCTCCGGCGTGCACACC TTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTAC AGCCTGAGCAGCGTCGTGACTGTGCCTAGTTCCA GCCTGGGCACCCAGACCTATATCTGCAACGTGAA GCCCAGCAACACCAAGGTGGACAAGAGA GTGGAGCCCAAGAGCTGCGACAAGACCCACACC CCCTGCCCAGCTCCAGAACTGCTGGGA GGACCCAGCGTGTTCCTGTTCCCCCCCAAGCCCA AGGACACCCTGATGATCAGCAGGACCCCCGAGG TGACCTGCGTGGTGGTGGACGTGTCCCACGAGG ACCCAGAGGTGAAGTTCAACTGGTACGTGGACGG GGTGGAGGTGCACAACGCCAAGACCAAGCCCAG AGAGGAGCAGTACAACAGCACCTACAGGGTGGT CCTGACAGTGCTGCACCAGGATTGGCTG AACGGCAAAGAATACAAGTGCAAAGTCTCCAACA AGGCCCTGCCAGCCCCAATCGAAAAGACAATCAG CAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGT GTACACCCTGCCCCCCAGCCGGGAGGAGATGAC CAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAG GGCTTCTACCCCAGCGATATCGCCGTGGAGTGG W0 2014/122613 1 64 AACGGCCAGCCCGAGAACAACTACAAG ACCACCCCCCCAGTGCTGGACAGCGACGGCAGC TTCTTCCTGTACAGCAAGCTGACCGTGGACAAGT CCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCA GCGTGATGCACGAGGCCCTGCACAACCACTACAC CCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG ALTERNAHVEPN GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGA ENCODWKBSEQID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NO:&3 TAGACCTAGCCAGGGAATCTACTGGGAGCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG CTGCTGATCTACGACGCTAGTAGTCTGGAACAGG GCGTGCCCTCTAGGTTTNSCGGCTCAGGCTCAGG CACCGACTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TTAACTCCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCC AGCGTGTTCKKNTCCCCCCCAGCGACGAGCAGC TGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGC TGAACAACTTCTACCCCCGGGAGGCCAAGGTGCA GTGGAAGGTGGACAACGCCCTGCAGAGCGGCAA CAGCCAGGAGAGCGTCACCGAGCAGGACAGCAA GGACTCCACCTACAGCCTGAGCAGCACCCTGACC CTGAGCAAGGCCGACTACGAGAAGCATAAGGTGT ACGCCTGCGAGGTGACCCACCAGGGCCTGTCCA GCCCCGTGACCAAGAGCTTCAACAGGGGCGAGT XAB4, CDRH1 _\ A GFTFSSY XAB4, CDRH2 N (CHOTHIA) KQDGSE XAB4, CDRH3 OD CHOTHIA DRGSLYY XAB4, CDRL1 (CHOTHIA) SQGHMNE XAB4, CDRL2 01 (CHOTHIA) UA(I) XAB4, CDRL3 (CHOTHIA) FNSYPL XAB4, CDRH1 \l (KABAT) SYWWB XAB4, CDRH2 (KABAT) MKQDGSEKYYVDSVKG XAB4, CDRH3 OD (KABAT) Y XAB4, CDRL1 (KABAT) RPSQGINWELA XAB4, CDRL2 (KABAT) DASSLEQ XAB4, CDRL3 KABAT PLT XAB4, VH _\ N EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS PGKGLEVWHUMKQDGSEKYYVDSVKGRFW SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY VWSQGTLVTVSS 1 65 XAB4, VL 43 AIQLTQSPSSLSASVGDRVTITCRPSQGINWELAVW QQKPGKAPKLLIYDASSLEQGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK XAB4, HEAVY CHAIN 4 EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLM ISRTPEVTCVVVDVSHEDPEVKFNVWVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK XAB4, LIGHT CHAIN 44 SPSSLSASVGDRVTITCRPSQGINWELAVW APKLLIYDASSLEQGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRT VAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC PN ENCODING SEQ 16 GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 12 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGTACTACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA GGACACCGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACTATTGGGGCCAGGGCACCCT GGTCACCGTGTCCAGC PN ENCODING SEQ 45 GCCATCCAGCTGACCCAGAGCCCCAGCAGCCTG ID NO: 43 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC TGTCGGCCCAGCCAGGGCATCAACTGGGAGCTG GCCTGGTATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAC AGGGCGTGCCCAGCCGGTTCAGCGGCAGCGGAT CCGACTTCACCCTGACCATCAGCTCCCT CGAGGACTTCGCCACCTACTACTGCCAG CAGTTCAACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAG PN ENCODING SEQ 8 GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 14 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGTACTACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA CGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACTATTGGGGCCAGGGCACCCT GGTCACCGTGTCCAGCGCTAGCACCAAGGGCCC CAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAG CACCAGCGGCGGCACAGCCGCCCTGGGCTGCCT GGTGAAGGACTACTTCCCCGAGCCCGTGACCGT GTCCTGGAACAGCGGAGCCCTGACCTCCGGCGT CTTCCCCGCCGTGCTGCAGAGCAGCGG CCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCC AGCAGCAGCCTGGGCACCCAGACCTACATCTGCA ACGTGAACCACAAGCCCAGCAACACCAAGGTGGA CAAGAGAGTGGAGCCCAAGAGCTGCGACAAGAC CCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCC AAGGACACCCTGATGATCAGCAGGACCC CCGAGGTGACCTGCGTGGTGGTGGACGTGAGCC ACGAGGACCCAGAGGTGAAGTTCAACTGGTACGT GGACGGCGTGGAGGTGCACAACGCCAAGACCAA GCCCAGAGAGGAGCAGTACAACAGCACCTACAG GGTGGTGTCCGTGCTGACCGTGCTGCACCAGGA CTGGCTGAACGGCAAGGAATACAAGTGCAAGGTC TCCAACAAGGCCCTGCCAGCCCCCATCGAAAAGA GCAAGGCCAAGGGCCAGCCACGGGAGC CCCAGGTGTACACCCTGCCCCCCTCCCGGGAGG AGATGACCAAGAACCAGGTGTCCCTGACCTGTCT GGTGAAGGGCTTCTACCCCAGCGACATCGCCGT GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAA CTACAAGACCACCCCCCCAGTGCTGGACAGCGAC GGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGG ACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCA GCTGCAGCGTGATGCACGAGGCCCTGCACAACC ACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGG CAAG PN ENCODING SEQ GCCATCCAGCTGACCCAGAGCCCCAGCAGCCTG 1 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC TGTCGGCCCAGCCAGGGCATCAACTGGGAGCTG GCCTGGTATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAC AGGGCGTGCCCAGCCGGTTCAGCGGCAGCGGAT CCGACTTCACCCTGACCATCAGCTCCCT GCAGCCCGAGGACTTCGCCACCTACTACTGCCAG CAGTTCAACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAGCGTACGGTGGCCG CTCCCAGCGTGTTCKKNTCCCCCCCAGCGACGA GCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTG CCTGCTGAACAACTTCTACCCCCGGGAGGCCAAG GTGCAGTGGAAGGTGGACAACGCCCTGCAGAGC AGCCAGGAGAGCGTCACCGAGCAGGAC AGCAAGGACTCCACCTACAGCCTGAGCAGCACCC TGACCCTGAGCAAGGCCGACTACGAGAAGCATAA GGTGTACGCCTGCGAGGTGACCCACCAGGGCCT GTCCAGCCCCGTGACCAAGAGCTTCAACAGGGG CGAGTGC ALTERNATIVE PN GAGGTGCAGCTGGTGGAATCAGGAGGCGACCTG ENCODING SEQ ID GTGCAGCCTGGCGGCTCACTGAGACTGAGCTGC NOVQ GCCGCTAGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC TGGAGTGGGTGGCCAAUUTAAGCAGGACGGCTC AGAGAAGTACTACGTGGACTCAGTGAAGGGCCG WO 22613 GTTCACTATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCT ALTERNATIVE PN GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGA ENCODING SEQ ID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NO:43 TAGACCTAGTCAGGGGATTAACTGGGAGCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG CTGCTGATCTACGACGCTAGTAGTCTGGAACAGG GCGTGCCCTCTAGGTTTNSCGGCTCAGGCTCAGG CACCGACTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TTAACTCCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAG ALTERNATIVE PN GAGGTGCAGCTGGTGGAATCAGGAGGCGACCTG ENCODING SEQ ID GTGCAGCCTGGCGGCTCACTGAGACTGAGCTGC NO:14 GCCGCTAGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC TGGAGTGGGTGGCCAAUUTAAGCAGGACGGCTC GTACTACGTGGACTCAGTGAAGGGCCG TATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCTGCTAGCACCAAGGGCCCAAGTG TCTTTCCCCTGGCCCCCAGCAGCAAGTCCACAAG CACTGCAGCTCTGGGTTGTCTGGTGAA GGACTACTTCCCCGAGCCCGTGACAGTGTCCTGG AACAGCGGAGCCCTGACCTCCGGCGTGCACACC GCCGTGCTGCAGAGCAGCGGCCTGTAC AGCCTGAGCAGCGTCGTGACTGTGCCTAGTTCCA GCCTGGGCACCCAGACCTATATCTGCAACGTGAA CCACAAGCCCAGCAACACCAAGGTGGACAAGAGA GTGGAGCCCAAGAGCTGCGACAAGACCCACACC TGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGA AGCGTGTTCCTGTTCCCCCCCAAGCCCA AGGACACCCTGATGATCAGCAGGACCCCCGAGG TGACCTGCGTGGTGGTGGACGTGTCCCACGAGG ACCCAGAGGTGAAGTTCAACTGGTACGTGGACGG GGTGGAGGTGCACAACGCCAAGACCAAGCCCAG AGAGGAGCAGTACAACAGCACCTACAGGGTGGT GTCCGTCCTGACAGTGCTGCACCAGGATTGGCTG AACGGCAAAGAATACAAGTGCAAAGTCTCCAACA AGGCCCTGCCAGCCCCAATCGAAAAGACAATCAG CAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGT GTACACCCTGCCCCCCAGCCGGGAGGAGATGAC CAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAG GGCTTCTACCCCAGCGATATCGCCGTGGAGTGG GAGAGCAACGGCCAGCCCGAGAACAACTACAAG ACCACCCCCCCAGTGCTGGACAGCGACGGCAGC TTCTTCCTGTACAGCAAGCTGACCGTGGACAAGT CCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCA GCGTGATGCACGAGGCCCTGCACAACCACTACAC CCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG 1 68 ALTERNATIVE PN GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGA ENCODING SEQ ID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NO:44 TAGACCTAGTCAGGGGATTAACTGGGAGCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG CTGCTGATCTACGACGCTAGTAGTCTGGAACAGG GCGTGCCCTCTAGGTTTNGCGGCTCAGGCTCAGG CACCGACTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TTAACTCCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCC AGCGTGTTCAKNTCCCCCCCAGCGACGAGCAGC TGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGC TGAACAACTTCTACCCCCGGGAGGCCAAGGTGCA GTGGAAGGTGGACAACGCCCTGCAGAGCGGCAA CAGCCAGGAGAGCGTCACCGAGCAGGACAGCAA GGACTCCACCTACAGCCTGAGCAGCACCCTGACC CTGAGCAAGGCCGACTACGAGAAGCATAAGGTGT ACGCCTGCGAGGTGACCCACCAGGGCCTGTCCA GCCCCGTGACCAAGAGCTTCAACAGGGGCGAGT SECOND GAGGTGCAGCTGGTGGAATCTGGCGGCGACCTG ALTERNATIVE PN GTGCAGCCTGGCGGCTCTCTGAGACTGTCTTGCG ENCODING SEQ ID CCGCCTCCGGCTTCACCTTCTCCAGCTACTGGAT NO:12 GTCCTGGGTGCGACAGGCCCCTGGCAAGGGACT GGTGGCCAACATCAAGCAGGACGGCTC CGAGAAGTACTACGTGGACTCCGTGAAGGGCCG GTTCACCATCTCCCGGGACAACGCCAAGAACTCC CTGTACCTGCAGATGAACTCCCTGCGGGCCGAG GACACCGCCGTGTACTACTGCGCCAGGGACCGG GGCTCCCTGTACTATTGGGGCCAGGGCACCCTG GTGACAGTGTCCTCC SECOND GCCATCCAGCTGACCCAGTCCCCCTCCAGCCTGT ALTERNATIVE PN CTGCCTCCGTGGGCGACAGAGTGACCATCACCTG ENCODING SEQ ID TCGGCCCTCCCAGGGCATCAACTGGGAACTGGC NO: 43 TCAGCAGAAGCCCGGCAAGGCCCCCAA GATCTACGACGCCAGCTCCCTGGAACAG GGCGTGCCCTCCAGATTCTCCGGCTCTGGCTCCG GCACCGACTTCACCCTGACCATCTCCAGCCTGCA GCCCGAGGACTTCGCCACCTACTACTGCCAGCAG TCCTACCCCCTGACCTTCGGCGGAGGCA CCAAGGTGGAAATCAAG SECOND GAGGTGCAGCTGGTGGAATCTGGCGGCGACCTG ALTERNATIVE PN CCTGGCGGCTCTCTGAGACTGTCTTGCG ENCODING SEQ ID CCGCCTCCGGCTTCACCTTCTCCAGCTACTGGAT NO: 14 GTCCTGGGTGCGACAGGCCCCTGGCAAGGGACT GGAATGGGTGGCCAACATCAAGCAGGACGGCTC CGAGAAGTACTACGTGGACTCCGTGAAGGGCCG GTTCACCATCTCCCGGGACAACGCCAAGAACTCC CTGTACCTGCAGATGAACTCCCTGCGGGCCGAG GACACCGCCGTGTACTACTGCGCCAGGGACCGG GGCTCCCTGTACTATTGGGGCCAGGGCACCCTG GTGACAGTGTCCTCCGCCTCCACCAAGGGCCCAA GCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCA CCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGG TGAAGGACTACTTCCCCGAGCCCGTGACCGTGTC W0 2014/122613 CTGGAACAGCGGAGCCCTGACCTCCGGCGTGCA CACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCT GTACAGCCTGAGCAGCGTGGTGACCGTGCCCAG CAGCAGCCTGGGCACCCAGACCTACATCTGTAAC GTGAACCACAAGCCCAGCAACACCAAGGTGGACA AGAGAGTGGAGCCCAAGAGCTGTGACAAGACCC ACACCTGCCCCCCCTGCCCAGCCCCCGAGCTGC TGGGCGGACCCAGCGTGTTCCTGTTCCCCCCCAA GCCCAAGGACACCCTGATGATCAGCAGAACCCCC GAGGTGACCTGTGTGGTGGTGGACGTGTCCCAC GAGGACCCAGAGGTGAAGTTCAACTGGTACGTGG ACGGCGTGGAGGTGCACAACGCCAAGACCAAGC CCAGAGAGGAGCAGTACAACAGCACCTACAGGGT GGTGTCCGTGCTGACCGTGCTGCACCAGGACTG GCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCC AACAAGGCCCTGCCAGCCCCAATCGAAAAGACCA TCAGCAAGGCCAAGGGCCAGCCAAGAGAGCCCC AGGTGTACACCCTGCCACCCAGCAGGGAGGAGA TGACCAAGAACCAGGTGTCCCTGACCTGTCTGGT GAAGGGCTKHACCCAAGCGACATCGCCGTGGA GTGGGAGAGCAACGGCCAGCCCGAGAACAACTA CAAGACCACCCCCCCAGTGCTGGACAGCGACGG CAGCTTCTTCCTGTACAGCAAGCTGACCGTGGAC AAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCT GCTCCGTGATGCACGAGGCCCTGCACAACCACTA CACCCAGAAGAGCCTGAGCCTGTCCCCAGGCAA SECOND 52 GCCATCCAGCTGACCCAGTCCCCCTCCAGCCTGT ALTERNAHVEPN CTGCCTCCGTGGGCGACAGAGTGACCATCACCTG ENCODWflSSEQID TCGGCCCTCCCAGGGCATCAACTGGGAACTGGC NO:¢1 CTGGTATCAGCAGAAGCCCGGCAAGGCCCCCAA GCTGCTGATCTACGACGCCAGCTCCCTGGAACAG GGCGTGCCCTCCAGATTCTCCGGCTCTGGCTCCG GCACCGACTTCACCCTGACCATCTCCAGCCTGCA GGACTTCGCCACCTACTACTGCCAGCAG TTCAACTCCTACCCCCTGACCTTCGGCGGAGGCA CCAAGGTGGAAATCAAGCGTACGGTGGCCGCTC CCAGCGTGTTCATCTTCCCCCCAAGCGACGAGCA GAGCGGCACCGCCAGCGTGGTGTGTCT GCTGAACAACTTCTACCCCAGGGAGGCCAAGGTG AAGGTGGACAACGCCCTGCAGAGCGGC AACAGCCAGGAGAGCGTCACCGAGCAGGACAGC AAGGACTCCACCTACAGCCTGAGCAGCACCCTGA CCCTGAGCAAGGCCGACTACGAGAAGCACAAGG CCTGTGAGGTGACCCACCAGGGCCTGTC CAGCCCCGTGACCAAGAGCTTCAACAGGGGCGA GTGC (CHOTHIA) (CHOTHIA) (CHOTHIA) WO 22613 _-—CHOTHIA DAS —_—(CHOTHIA) FNSYPL —-—KABAT SYWMS —_—(KABAT) NIKQDGSEKYYVDSVKG —-—(KABAT) DRGSLYY —-—(KABAT) NWELA —-—(KABAT) N BAT) QQFNSYPLT EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY VTVSS XAB5, VL AIQLTQSPSSLSASVGDRVTITCRPSQGINWELAVW QQKPGKAPKLLIYDASSLENGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK XAB5, HEAVY CHAIN EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMS VWRQAPGKGLEVWANIKQDGSEKYYVDSVKGRFTI SRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNVWVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK XAB5, LIGHT CHAIN 54 AIQLTQSPSSLSASVGDRVTITCRPSQGINWELAVW QQKPGKAPKLLIYDASSLENGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC PN ENCODING SEQ 16 GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG ID NO: 12 GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGTACTACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA GGACACCGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACTATTGGGGCCAGGGCACCCT GGTCACCGTGTCCAGC PN ENCODING SEQ—55 GCCATCCAGCTGACCCAGAGCCCCAGCAGCCTG 3 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC TGTCGGCCCAGCCAGGGCATCAACTGGGAGCTG TATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAA ACGGCGTGCCCAGCCGGTTCAGCGGCAGCGGAT CCGGCACCGACTTCACCCTGACCATCAGCTCCCT GCAGCCCGAGGACTTCGCCACCTACTACTGCCAG CAGTTCAACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAG PN ENCODING SEQ GAGGTGCAGCTGGTCGAGTCTGGCGGCGACCTG IDNOVM GTGCAGCCTGGCGGCAGCCTGAGACTGAGCTGC GCCGCCAGCGGCTTCACCTTCAGCAGCTACTGGA TGTCCTGGGTCCGCCAGGCCCCTGGCAAAGGCC TCGAATGGGTGGCCAACATCAAGCAGGACGGCA GCGAGAAGDMNACGTGGACAGCGTGAAGGGCC GGTTCACCATCAGCCGGGACAACGCCAAGAACAG CCTGTACCTGCAGATGAACAGCCTGCGGGCCGA CGCCGTGTACTACTGCGCCAGGGACCG GGGCAGCCTGTACUUTGGGGCCAGGGCACCCT GGTCACCGTGTCCAGCGCTAGCACCAAGGGCCC CAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAG CACCAGCGGCGGCACAGCCGCCCTGGGCTGCCT GGTGAAGGACTACTTCCCCGAGCCCGTGACCGT GAACAGCGGAGCCCTGACCTCCGGCGT GCACACCTTCCCCGCCGTGCTGCAGAGCAGCGG CCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCC AGCAGCAGCCTGGGCACCCAGACCTACATCTGCA ACGTGAACCACAAGCCCAGCAACACCAAGGTGGA AGTGGAGCCCAAGAGCTGCGACAAGAC CCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCC AAGCCCAAGGACACCCTGATGATCAGCAGGACCC CCGAGGTGACCTGCGTGGTGGTGGACGTGAGCC ACGAGGACCCAGAGGTGAAGTTCAACTGGTACGT GGACGGCGTGGAGGTGCACAACGCCAAGACCAA GCCCAGAGAGGAGCAGTACAACAGCACCTACAG GTCCGTGCTGACCGTGCTGCACCAGGA CTGGCTGAACGGCAAGGAATACAAGTGCAAGGTC TCCAACAAGGCCCTGCCAGCCCCCATCGAAAAGA CCATCAGCAAGGCCAAGGGCCAGCCACGGGAGC CCCAGGTGTACACCCTGCCCCCCTCCCGGGAGG AGATGACCAAGAACCAGGTGTCCCTGACCTGTCT GGTGAAGGGCTTCTACCCCAGCGACATCGCCGT GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAA CTACAAGACCACCCCCCCAGTGCTGGACAGCGAC GGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGG ACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCA GCTGCAGCGTGATGCACGAGGCCCTGCACAACC ACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGG CAAG PN ENCODING SEQ GCCATCCAGCTGACCCAGAGCCCCAGCAGCCTG |DNO:&1 AGCGCCAGCGTGGGCGACAGAGTGACCATCACC TGTCGGCCCAGCCAGGGCATCAACTGGGAGCTG GCCTGGTATCAGCAGAAGCCTGGCAAGGCCCCC AAGCTGCTGATCTACGACGCCAGCTCCCTGGAAA 1 72 ACGGCGTGCCCAGCCGGTTCAGCGGCAGCGGAT CCGGCACCGACTTCACCCTGACCATCAGCTCCCT GCAGCCCGAGGACTTCGCCACCTACTACTGCCAG CAGTTCAACAGCTACCCCCTGACCTTCGGCGGAG GCACCAAGGTGGAAATCAAGCGTACGGTGGCCG CTCCCAGCGTGTTCAKNTCCCCCCCAGCGACGA GCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTG CCTGCTGAACAACTTCTACCCCCGGGAGGCCAAG GTGCAGTGGAAGGTGGACAACGCCCTGCAGAGC GGCAACAGCCAGGAGAGCGTCACCGAGCAGGAC AGCAAGGACTCCACCTACAGCCTGAGCAGCACCC TGACCCTGAGCAAGGCCGACTACGAGAAGCATAA GGTGTACGCCTGCGAGGTGACCCACCAGGGCCT CCCCGTGACCAAGAGCTTCAACAGGGG ALTERNATIVE PN GAGGTGCAGCTGGTGGAATCAGGAGGCGACCTG ENCODING SEQ ID GTGCAGCCTGGCGGCTCACTGAGACTGAGCTGC NO: 12 GCCGCTAGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC GGGTGGCCAAUUTAAGCAGGACGGCTC AGAGAAGTACTACGTGGACTCAGTGAAGGGCCG GTTCACTATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCT ALTERNATIVE PN GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGA ENCODING SEQ ID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NO: 53 TAGACCTAGTCAGGGGATTAACTGGGAGCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG CTGCTGATCTACGACGCTAGTAGTCTGGAAAACG GCGTGCCCTCTAGGTTTNSCGGCTCAGGCTCAGG CACCGACTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TTAACTCCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAG ATIVE PN CAGCTGGTGGAATCAGGAGGCGACCTG ENCODING SEQ ID GTGCAGCCTGGCGGCTCACTGAGACTGAGCTGC NO: 14 GCCGCTAGTGGCTTCACCTTTAGTAGCTACTGGA TGAGCTGGGTGCGACAGGCCCCTGGCAAGGGAC TGGAGTGGGTGGCCAAUUTAAGCAGGACGGCTC AGAGAAGTACTACGTGGACTCAGTGAAGGGCCG GTTCACTATTAGCCGGGATAACGCTAAGAATAGC CTGTACCTGCAGATGAATAGCCTGAGAGCCGAGG ACACCGCCGTGTACTACTGCGCTAGAGATAGAGG CTCACTGTACTACTGGGGCCAGGGCACCCTGGTG ACAGTGTCTTCTGCTAGCACCAAGGGCCCAAGTG CCCTGGCCCCCAGCAGCAAGTCCACAAG CGGAGGCACTGCAGCTCTGGGTTGTCTGGTGAA GGACTACTTCCCCGAGCCCGTGACAGTGTCCTGG AACAGCGGAGCCCTGACCTCCGGCGTGCACACC TTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTAC AGCCTGAGCAGCGTCGTGACTGTGCCTAGTTCCA GCCTGGGCACCCAGACCTATATCTGCAACGTGAA CCACAAGCCCAGCAACACCAAGGTGGACAAGAGA GTGGAGCCCAAGAGCTGCGACAAGACCCACACC TGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGA GGACCCAGCGTGTTCCTGTTCCCCCCCAAGCCCA AGGACACCCTGATGATCAGCAGGACCCCCGAGG TGACCTGCGTGGTGGTGGACGTGTCCCACGAGG ACCCAGAGGTGAAGTTCAACTGGTACGTGGACGG GGTGGAGGTGCACAACGCCAAGACCAAGCCCAG AGAGGAGCAGTACAACAGCACCTACAGGGTGGT CCTGACAGTGCTGCACCAGGATTGGCTG AACGGCAAAGAATACAAGTGCAAAGTCTCCAACA AGGCCCTGCCAGCCCCAATCGAAAAGACAATCAG CAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGT GTACACCCTGCCCCCCAGCCGGGAGGAGATGAC CAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAG GGCTTCTACCCCAGCGATATCGCCGTGGAGTGG GAGAGCAACGGCCAGCCCGAGAACAACTACAAG ACCACCCCCCCAGTGCTGGACAGCGACGGCAGC TTCTTCCTGTACAGCAAGCTGACCGTGGACAAGT CCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCA TGCACGAGGCCCTGCACAACCACTACAC CCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG ATIVE PN 58 GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGA ENCODING SEQ ID GCGCTAGTGTGGGCGATAGAGTGACTATCACCTG NO: 54 TAGACCTAGTCAGGGGATTAACTGGGAGCTGGCC TGGTATCAGCAGAAGCCCGGCAAGGCCCCTAAG ATCTACGACGCTAGTAGTCTGGAAAACG GCGTGCCCTCTAGGTTTNGCGGCTCAGGCTCAGG CACCGACTTCACCCTGACTATTAGTAGCCTGCAG CCCGAGGACTTCGCTACCTACTACTGTCAGCAGT TTAACTCCTACCCCCTGACCTTCGGCGGAGGCAC TAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCC AGCGTGTTCKKNTCCCCCCCAGCGACGAGCAGC TGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGC TGAACAACTTCTACCCCCGGGAGGCCAAGGTGCA GTGGAAGGTGGACAACGCCCTGCAGAGCGGCAA CAGCCAGGAGAGCGTCACCGAGCAGGACAGCAA GGACTCCACCTACAGCCTGAGCAGCACCCTGACC CTGAGCAAGGCCGACTACGAGAAGCATAAGGTGT ACGCCTGCGAGGTGACCCACCAGGGCCTGTCCA GCCCCGTGACCAAGAGCTTCAACAGGGGCGAGT LEADERSEQUENCE MEFGLSWVFLVAI LEGVHC OFTHEHEAVY CHNN SEQUENCE MDMRVPAQLLGLLLLWLPGARC OFTHEUGHT CHNN PN ENCODING SEQ 1 ATGGAATTCGGCCTGAGCTGGGTGTTCCTGGTCG IDNCt59 CGATTCTGGAAGGCGTGCACTGC PN ENCODING SEQ ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGG IDNO:&) CTTCTGCTGCTCTGGCTCCCAGGCGCCAGATGT ALTERNAHVE 3 IPFLMAAAQSVQA LEADERSEQUENCE OFTHEHEAVY W0 2014/122613 1 74 ALTERNATIVE 64 MSVLTQVLALLLLWLTGTRC LEADER SEQUENCE OF THE LIGHT CHAIN ALTERNATIVE PN 65 ATGGCCTGGGTGTGGACCCTGCCCTTCCTGATGG ENCODING SEQ ID CTCAGTCAGTGCAGGCC NO: 63 ALTERNATIVE PN ATGAGCGTGCTGACTCAGGTGCTGGCCCTGCTGC NG SEQ ID TGCTGTGGCTGACCGGCACCCGCTGC NO: 64 SECOND MEWSVWFLFFLSVTTGVHS ALTERNATIVE LEADER SEQUENCE OF THE HEAVY CHAIN SECOND MSVPTQVLGLLLLWLTDARC ALTERNATIVE LEADER SEQUENCE OF THE LIGHT CHAIN SECOND ATGGAATGGTCCTGGGTGTTCCTGTTCTTCCTGTC ATIVE PN CGTGACCACAGGCGTGCACTCC ENCODING SEQ ID NO: 67 SECOND ATGTCCGTGCCCACACAGGTGCTGGGCCTGCTG ALTERNATIVE PN CTGCTGTGGCTGACCGACGCCAGATGC ENCODING SEQ ID NO: 68 71 SQX1IX2X3X4 CDRL1 (CHOTHIA) 72 FX1SYPL CDRL3 CHOTHIA CONSENSUS, 73 RPSQX1IX2X3X4LA CDRL1 (KABAT) CONSENSUS, DASSLEX1 CDRL2 KABAT CONSENSUS, QQFX1SYPLT CDRL3 (KABAT) I7A 76 GITIPRNPGCPNSEDKNFPRTVMVNLNIHNRNTNTN YYNRSTSPWNLHRNEDPERYPSVIWEAKC RH LGCI NADGNVDYHMNSVPIQQEI LVLRREPPHCP NSFRLEKILVSVGCTCVTPIVHHVAEFRH huIL-17F 77 MRKIPKVGHTFFQKPESCPPVPGGSMKLDIGIINEN QRVSMSRNI ESRSTSPWNYTVTWDPNRYPSEVVQ AQCRNLGCINAQGKEDISMNSVPIQQETLVVRRKHQ GCSVSFQLEKVLVTVGCTCVTPVIHHVQ alternative huIL-17A 78 GPIVKAGITI PRNPGCPNSEDKNFPRTVMVNLNIHNR KRSSDYYNRSTSPWNLHRNEDPERYPSVI WEAKCRHLGCI NADGNVDYHMNSVPIQQEI LVLRRE PPHCPNSFRLEKILVSVGCTCVTPIVHHVA cynolL-17A 79 GIAIPRNSGCPNSEDKNFPRTVMVNLNIHNRNTSTN PKRSSDYYNRSTSPWNLHRNEDPERYPSVIWEAKC RHLGCVKADGNVDYHMNSVPIQQEILVLRREPRHC W0 2014/122613 1 75 PNSFRLEKILVSVGCTCVTPIVHHVA cynoIL-17F MRKI PKVGHTFFQKPESCPPVPEGSMKLDTGIINEN QRVSMSRN| ESRSTSPWNYTVTWDPNRYPSEVVQ AQCKHLGCI NAQGKEDISMNSVPIQQETLVLRRKHQ GCSVSFQLEKVLVTVGCTCVTPVVHHVQ rhesusIL-17A O) _\ GIAI PRNSGCPNSEDKNFPRTVMVNLNIHNRNTSTS PKRSSDYYNRSTSPWNLHRNEDPERYPSVIWEAKC RHLGCVKADGNVDYHMNSVPIQQEILVLRREPRHC PNSFRLEKILVSVGCTCVTPIVHHVA marmosetIL-17A O)N CPNAEDKNFPRTVMVNLNIRNRNTNSKRA SDYYNRSSSPWNLHRNEDPERYPSVIWEAKCRHLG CVDADGNVDYHMNSVPIQQEILVLRREPRHCTNSF RLEKMLVSVGCTCVTPIVHHVA O) 00 MAAI | PQSSACPNTEAKDFLQNVKVNLKVFNSLGAK VSSRRPSDYLNRSTSPVVTLHRNEDPDRYPSVIWEA QCRHQRCVNAEGKLDHHMNSVLIQQEI LVLKREPES CPFTFRVEKMLVGVGCTCVASIVRQAA mlL—17F 0) RHRKNPKAGVPALQKAGNCPPLEDNTVRV DI RI FNQNQGISVPREFQNRSSSPWDYNITRDPHRF PSEIAEAQCRHSGCI NAQGQEDSTMNSVAIQQEI LV LRREPQGCSNSFRLEKMLLKVGCTCVKPIVHQAA ratIL-17A 0) 0| MAVLIPQSSVCPNAEANNFLQNVKVNLKVI NSLSSK ASSRRPSDYLNRSTSPVVTLSRNEDPDRYPSVIWEA QCRHQRCVNAEGKLDHHMNSVLIQQEI LVLKREPEK VEKMLVGVGCTCVSSIVRHAS hulL—17RA NCTVKNSTCLDDSWI HPRNLTPSSPKDLQIQLHFAH FPVAH| EVVTLQTDASI LYLEGAELSVLQLNT NERLCVRFEFLSKLRHHHRRWRFTFSHFVVDPDQE YEVTVHHLPKPI PDGDPNHQSKNFLVPDCEHARMK SSGSLWDPNITVETLEAHQLRVSFTLWNES THYQI LLTSFPHMENHSCFEHMHHIPAPRPEEFHQR SNVTLTLRNLKGCCRHQVQIQPFFSSCLNDCLRHSA TVSCPEMPDTPEPIPDYMPLWEFRHDSGGGLNDI F EAQKIEWHE

Claims (18)

CLAIMS is:

1. An isolated antibody or protein comprising an antigen-binding portion f, comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein: a) said VH comprises, in sequence, the three complementarity determining regions (CDRs) set forth in SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 3, b) said VL comprises, in sequence, the three CDRs set forth in SEQ ID NO: 42, SEQ ID NO: 23 and SEQ ID NO: 11, and wherein said antibody or protein specifically binds to homodimeric IL-17A and heterodimeric IL-17AF, but does not specifically bind to homodimeric IL-17F.

2. An isolated antibody or n according to claim 1, wherein the IL-17A, IL- 17AF or IL-17F are selected from one or more of lgus monkey, rhesus macaque monkey, marmoset monkey, rat, mouse or human.

3. An isolated antibody or protein according to claim 1 or 2, comprising the VH amino acid sequence set forth in SEQ ID NO: 12, and the VL amino acid sequence set forth in SEQ ID NO: 43.

4. An isolated antibody or protein according any one of the preceding claims, comprising the amino acid ce set forth in SEQ ID NO: 14, and the amino acid sequence set forth in SEQ ID NO: 44.

5. An isolated antibody or protein according to any one of the ing claims, which is conjugated to a further active .

6. An isolated antibody or protein according to any one of the preceding claims, which is a monoclonal antibody or an antigen-binding portion f.

7. An isolated antibody or protein according to claim 6, which is a ic, humanized, or human antibody or portion thereof.

8. A pharmaceutical composition comprising an dy or protein according to anyone of the preceding claims, in combination with one or more pharmaceutically acceptable excipient, diluent or carrier.

9. A pharmaceutical composition according to claim 8, further comprising one or more additional active ingredients.

10. Use of an antibody or protein according to any one of claims 1 to 7 in the manufacture of a medicament for the ent of a pathological disorder mediated by IL-17A or that can be treated by ting IL-6 or GRO-alpha secretion.

11. The use according to claim 10, wherein the pathological disorder is an inflammatory disorder or condition.

12. The use according to claim 11, wherein the inflammatory disorder or condition is arthritis, rheumatoid arthritis, psoriasis, chronic obstructive pulmonary disease, systemic lupus erythematosus (SLE), lupus nephritis, asthma, multiple sclerosis or cystic fibrosis.

13. An isolated nucleic acid molecule encoding any one of the antibodies or proteins as defined in any one of claims 1 to 7.

14. A cloning or expression vector comprising one or more nucleic acid sequences according to claim 13, wherein the vector is suitable for the inant production of ed antibody or protein as defined by any one of claims 1 to 7.

15. A host cell sing one or more cloning or expression vectors according to claim 14, provided that if the cell is a human cell it is ex vivo.

16. An isolated nucleic acid molecule ing to claim 13, wherein the c acid le is messenger RNA (mRNA).

17. A process for the production of an isolated antibody or protein according to any one of claims 1 to 7, sing culturing a host cell according to claim 15, purifying and recovering said antibody or protein.

18. An isolated antibody or protein according to claim 1, substantially as herein bed with reference to any one of the Examples and/or