AU2006203778B2 - Therapeutic binding molecules - Google Patents

Description

P/00/0 II Regulation 3.2 AUSTRALIA Patents Act 1990 COMPLETE SPECIFICATION STANDARD PATENT (ORIGINAL) Name of Applicant: Novartis AG, of Lichtstrasse 35, Basel CH-4056, Switzerland Actual Inventors: Gregorio Aversa Frank Kolbinger Jose M. Carballido Herrera Andris Asz6di Josd W. Saldanha Bruce M. Hall Address for Service: DAVIES COLLISON CAVE, Patent & Trademark Attorneys, of 1 Nicholson Street, Melbourne, 3000, Victoria, Australia Ph: 03 9254 2777 Fax: 03 9254 2770 Attorney Code: DM Invention Title: "Therapeutic binding molecules" The following statement is a full description of this invention, including the best method of performing it known to us:- -1 Therapeutic binding molecules This application is a divisional of Australian Patent Application No. 2002231795, the entire contents of which is incorporated herein by reference. 5 Field of the Invention The present invention relates to organic compounds, such as to binding molecules against CD45 antigen isoforms, such as for example monoclonal antibodies (mAbs). 10 Background of the Invention One approach in the treatment of a variety of diseases is to achieve the elimination or the inactivation of pathogenic leukocytes and the potential for induction of tolerance to inactivate 15 pathological immune responses. Organ, cell and tissue transplant rejection and the various autoimmune diseases are thought to be primarily the result of T-cell mediated immune response triggered by helper T-cells which are capable of recognizing specific antigens which are captured, processed and 20 presented to the helper T cells by antigen presenting cell (APC) such as macrophages and dendritic cells, in the form of an antigen-MHC complex, I.e. the helper T-cell when recognizing specific antigens is stimulated to produce cytokines such as IL-2 and to express or upregulate some cytokine receptors and other activation molecules and to proliferate. Some of these activated helper T-cells may act directly or indirectly, I.e. assisting effector 25 cytotoxic T-ceils or B cells, to destroy cells or tissues expressing the selected antigen. After the termination of the immune response some of the mature clonally selected cells remain as memory helper and memory cytotoxic T-cells, which circulate in the body and rapidly recognize the antigen if appearing again. If the antigen triggering this response is an innocuous environmental antigen the result Is allergy, if the antigen Is not a foreign antigen, 30 but a self antigen, it can result is autoimmune disease; if the antigen Is an antigen from a transplanted organ, the result can be graft rejection. The immune system has developed to recognize self from non-self. This property enables an organism to survive In an environment exposed to the daily challenges of pathogens. This 35 specificity for non-self and tolerance towards self arises during the development of the T cell repertoire In the thymus through processes of positive and negative selection, which also comprise the recognition and elimination of autoreactive T cells. This type of tolerance Is -2 referred to as central tolerance. However, some of these autoreactive cells escape this selective mechanism and pose a potential hazard for the development of autoimmune diseases. To control the autoreactive T cells that have escaped to the periphery, the Immune system has peripheral reg ulatory mechanisms that provide protection against autoimmunity. 5 These mechanisms are a basis for peripheral tolerance. Cell surface antigens recognized by specific mAbs are generally designated by a CD (Cluster of Differentiation) number assigned by successive International Leukocyte Typing workshops and the term CD45 applied herein refers to the cell surface leukocyte common 10 antigen CD45; and an mAb to that antigen is designated herein as "anti-CD45". The leukocyte common antigen (LCA) or CD45 Is the major component of anti-lymphocyte globulin (ALG). CD45 belongs to the family of transmembrane tyrosine phosphat ases and is both a positive and negative regulator of cell activation, depending upon receptor interaction. 15 The phosphatase activity of CD45 appears to be required for activation of Src-family kinases associated with antigen receptor of B and T lymphocytes (Trowbridge IS et al, Annu Rev Immunol. 1994;12:85-116). Thus, in T cell activation, CD45 is essential for signal I and CD45-deficicient cells have profound defects in TCR-mediated activation events. 20 The CD45 antigen exists in different isoforms comprising a family of transmembrane glycoproteins. Distinct Isoforms of CD45 differ in their extracellular domain structure which arise from alternative splicing of 3 variable exons coding for part of the CD45 extracellular region (Streuli MF. et al, J. Exp. Med. 1987; 166:1548-1566). The various Isoforms of CD45 have different extra-cellular domains, but have the same transmembrane and cytoplasmic 25 segments having two homologous, highly conserved phosphatase domains of approximately 300 residues. Different isoform combinations are differentially expressed on subpopulations of T and B lymphocytes (Thomas ML. et al, Immunol. Today 1988; 9:320-325). Some monoclonal antibodies recognize an epitope common to all the different isoforms, while other mAbs have a restricted (CD45R) s pecificity, dependent on which of the alternatively spliced 30 exons (A, B or C) they recognize. For example, monoclonal antibodies recognizing the product of exon A are consequently designated CD45RA, those recognizing the various isoforms containing exon B have been designated CD45RB (Beverley PCL et al, Immunol. Supp. 1988; 1:3-5). Antibodies such as UCHL1 selectively bind to the 180 kDa isoform CD45RO (without any of the variable exons A, B or C) which appears to be restricted to a -3 subset of activated T cells, memory cells and cortical thymocytes and is not detected on B cells (Terry LA et al, Immunol. 1988; 64:331-336). Description of the Figures 5 Figure 1 shows that the inhibition of primary MLR by the "candidate mAb" is dose-dependent in the range of 0.001 and 10 pg/ml. "Concentration" is concentration of the "candidate mAb". Figure 2 shows the plasmid map of the expression vector HCMV-G1 HuAb-VHQ comprising the heavy chain having the nucleotide sequence SEQ ID NO:12 (3921-4274) in the complete 10 expression vector nucleotide sequence SEQ ID NO:15. Figure 3 shows the plasmid map of the expression vector HCMV-G1 HuAb-VHE comprising the heavy chain having the nucleotide sequence SEQ ID NO:1 1 (3921-4274) in the complete expression vector nucleotide sequence SEQ ID NO:16. Figure 4 shows the plasmid map of the expression vector HCMV-K HuAb-humV1 comprising 15 the light chain having the nucleotide sequence SEQ ID NO:14 (3964-4284) in the complete expression vector nucleotide sequence SEQ ID NO:17. Figure 5 shows the plasmid map of the expression vector HCMV-K HuAb-humV2 comprising the light chain having the nucleotide sequence SEQ ID NO:13 (3926-4246) in the complete expression vector nucleotide sequence SEQ ID NO:18. 20 Description of the Invention We have now found a binding molecule which comprises a polypeptide sequence which binds to CD45RO and CD45RB, hereinafter also designated as a "CD45RO/RB binding 25 molecule". These binding molecule according to the invention may induce immunosuppres sion, inhibit primary T cell responses and induce T cell tolerance. Furthermore, the binding molecules of the invention inhibit primary mixed lymphocyte responses (MLR). Cells derived from cultures treated with CD45RO/RB binding molecules preferredly also have impaired proliferative responses in secondary MLR even In the 30 absence of CD45RO/RB binding molecules In the secondary MLR. Such impaired proliferative responses In secondary MLR are an Indication of the ability of binding molecules of the Invention to Induce tolerance. Additionally, In vivo administration of CD45RO/RB binding molecule to severe combined Immunodeficiency (SCID) mice undergoing xeno GVHD following injection with human PBMC may prolong mice survival, compared to control -4 treated mice, even though circulating human T cells may still be detected in CD45RO/RB binding molecule treated mice. By "CD45RO/RB binding molecule" is meant any molecule capable of binding specifically to 5 the CD45RB and CD45RO isoforms of the CD45 antigen, either alone or associated with other molecules. The binding reaction may be shown by standard methods (qualitative assay) including for example any kind of binding assay such as direct or indirect immunofluorescence together with fluorescence microscopy or cytofluorimetric (FACS) analysis, enzyme-linked immunosorbent assay (ELISA) or radiolmmunoass ay in which 10 binding of the molecule to cells expressing a particular CD45 isoform can be visualized. In addition, the binding of this molecule may result in the alteration of the function of the cells expressing these isoforms. For example inhibition of primary or secondary mixed lymphocyte response (MLR) may be determined, such as an in vitro assay or a bloassay for determining the inhibition of primary or secondary MLR in the presence and in the absence of a 15 CD45RO/RB binding molecule and determining the differences in primary MLR inhibition. Alternatively, the in vitro functional modulatory effects can also be determined by measuring the PBMC or T cells or CD4' T cells proliferation, production of cytokines, change in the expression of cell surface molecules e.g. following cell activation in MLR, or following 20 stimulation with specific antigen such as tetanus toxoid or other antigens, or with polyclonal stimulators such as phytohemagglutinin (PHA) or anti-CD3 and anti-CD28 antibodies or phorbol esters and Ca 2 + ionophores. The cultures are set up in a similar manner as described for MLR except that instead of allogeneic cells as stimulators soluble antigen or polyclonal stimulators such as those mentioned above are used. T cell proliferation is 25 measured preferably as described above by 3 H-thymidine incorporation. Cytokine production is measured preferably by sandwich ELISA where a cytokine capture antibody is coated on the surface of a 96-well plate, the supematants from the cultures are added and incubated for I hr at room temperature and a detecting antibody specific for the particular cytokine is then added, following a second-step antibody conjugated to an enzyme 30 such as Horseradish peroxidase followed by the corresponding substrate and the absorbance is measured in a plate reader. The change in cell surface molecules may be preferably measured by direct or indirect immunofluorescence after staining the target cells with antibodies specific for a particular cell surface molecule. The antibody can be either directly labeled with flourochrome or a fluorescently labeled second step antibody specific for the first antibody can be used, and the cells are analysed with a cytofluorimeter. The binding molecule of the invention has a binding specificity for both CD45RO and 5 CD45RB ("CD45RB/RO binding molecule"). Preferably the binding molecule binds to CD45RO isoforms with a dissociation constant (Kd) <15nM, more preferably with a Kd<lOnM, most preferably with a Kd<5nM. Preferably the binding molecule binds binds to CD45RB isoforms with a Kd<15nM, more preferably with a Kd<lOnM, most preferably with a Kd<5nM. 10 In a further preferred embodiment the binding molecule of the invention binds those CD45 isoforms which 1) include the A and B epitopes but not the C epitope of the CD45 molecule; and/or 2) Include the B epitope but not the A and not the C epitope of the CD45 molecule; and/or 15 3) isoforms which do not include any of the A, B or C epitopes of the CD45 molecule. In yet a further preferred embodiment the binding molecule of the invention does not bind CD45 isoforms which include 1) all of the the A, B and C epitopes of the CD45 molecule; and/or 20 2) both the B and C epitopes but not the A epitope of the CD45 molecule. In further preferred embodiments the binding molecule of the invention further 1) recognises memory and in vivo alloactivated T cells; and/or 2) binds to its target on human T cells, such as for example PEER cells; wherein said 25 binding preferably is with a Kd<15nM, more preferably with a Kd<lOnM, most preferably with a Kd<5nM; and/or 3) Inhibits in vitro alloreactive T cell function, preferably with an IC5 of about 5nM, more preferably with an ICw of about I nM, most preferably with an IC5 of about 0,5nM or even 0,inM; and/or 30 4) induces alloantigen-specific T cell tolerance in vitro; and/or -6 5) prevents lethal xenogeneic graft versus host disease (GvHD) induced in SCID mice by injection of human PBMC when admistlered in an effective amount. In a further preferred embodiment the binding molecule of the invention binds to the same 5 epitope as the monoclonal antibody "A6" as described by Aversa et al., Cellular Immunology 158, 314-328 (1994). Due to the above-described binding properties and biological activities, such binding molecules of the invention are particularly useful in medicine, for therapy and/or prophylaxis. 10 Diseases in which binding molecules of the invention are particularly useful include autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies, as will be further set out below. We have found that a molecule comprising a polypeptide of SEQ ID NO: I and a polypeptide 15 of SEQ ID NO: 2 is a CD45RO/RB binding molecule. We also have found the hypervariable regions CDR1', CDR2' and CDR3' in a CD45RO/RB binding molecule of SEQ ID NO:1, CDR1' having the amino acid sequence Arg-Ala-Ser-Gin-Asn-Ile-Gly-Thr-Ser-lIe-GIn (RASQNIGTSIQ), CDR2' having the amino acid sequence Ser-Ser-Ser-Glu-Ser-lie-Ser (SSSESIS) and CDR3' having the amino acid sequence Gln-Gin-Ser-Asn-Thr-Trp-Pro-Phe 20 Thr (QQSNTWPFT). We also have found the hypervariable regions CDR1, CDR2 and C DR3 in a CD45RO/RB binding molecule of SEQ ID NO:2, CDR1 having the amino acid sequence Asn-Tyr-Ile-Ile His (NYIIH), CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His-Gly-Thr 25 Lys-Tyr-Asn-Glu-Lys-Phe -Lys-Gly (YFNPYNHGTKYNEKFKG) and CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-Ala-Trp-Phe-Asp-Thr (SGPYAWFDT). CDRs are 3 specific complementary determining regions which are also called hypervariable regions which essentially determine the antigen binding characteristics. These CDRs are 30 part of the variable region, e.g. of SEQ ID NO: 1 or SEQ ID NO: 2, respectively, wherein these CDRs alternate with framework regions (FR's) e.g. constant regions. A SEQ ID NO: 1 is part of a light chain, e.g. of SEQ ID NO: 3, and a SEQ ID NO:2 is part of a heavy chain, e.g. of SEQ ID NO: 4, in a chimeric antibody according to the present invention. The CDRs of a heavy chain together with the CDRs of an associated light chain essentially constitute -7 the antigen binding site of a molecule of the present Invention. It is known that the contribution made by a light chain variable region to the energetics of binding is small compared to that made by the associated heavy chain variable region and that isolated heavy chain variable regions have an antigen binding activity on their own. Such molecules 5 are commonly referred to as single domain antibodies. In one aspect the present invention provides a molecule comprising at least one antigen binding site, e.g. a CD45RO/RB binding molecule, comprising in sequence the hypervariable regions CDR1, CDR2 and CDR3, said CDR1 having the amino acid sequence Asn-Tyr-ile 10 lle-His (NYIIH), said CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His Gly-Thr-Lys-Tyr-Asn-Glu-Lys-Phe -Lys-Gly (YFNPYNHGTKYNEKFKG) and said CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-Aia-Trp-Phe-Asp-Thr (SGPYAWFDT); e.g. and direct equivalents thereof. 15 in another aspect the present invention provides a molecule comprising at least one antigen binding site, e.g. a CD45RO/RB binding molecule, comprising a) a first domain comprising in sequence the hypervariable regions CDR1, CDR2 and CDR3, said CDR1 having the amino acid sequence Asn-Tyr-Ile-Ile-His (NYIIH), said CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His-Gly-Thr-Lys-Tyr-Asn-Glu 20 Lys-Phe -Lys-Gly (YFNPYNHGTKYNEKFKG) and said CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-AJa-Trp-Phe-Asp-Thr (SGPYAWFDT); and b) a second domain comprising in sequence the hypervariable regions CDR1', CDR2' and CDR3, CDR1' having the amino acid sequence Arg-Ala-Ser-Gin-Asn-Ile-Gly-Thr-Ser-lie Gin (RASQNIGTSIQ), CDR2' having the amino acid sequence Ser-Ser-Ser-Glu-Ser-le 25 Ser (SSSESIS) and CDR3' having the amino acid sequence Gln-GIn-Ser-Asn-Thr-Trp Pro-Phe-Thr (QQSNTWPFT), e.g. and direct equivalents thereof. In a preferred embodiment the first domain comprising In sequence the hypervariable 30 regions CDR1, CDR2 and CDR3 is an immunoglobulin heavy chain, and the second domain comprising In sequence the hypervariable regions CDR1', CDR2' and CDR3' is an immunoglobulin light chain.

-8 In another aspect the present invention provides a molecule, e.g. a CD45RO/RB binding molecule, comprising a polypeptide of SEQ ID NO: I and/or a polypeptide of SEQ ID NO: 2. preferably comprising in one domain a polypeptide of SEQ ID NO: I and in another domain a polypeptide of SEQ ID NO: 2, e.g. a chimeric monoclonal antibody, and in another aspect 5 A molecule, e.g. a CD45RO/RB binding molecule, comprising a polypeptide of SEQ ID NO: 3 and/or a polypeptide of SEQ ID NO: 4, preferably comprising in one domain a polypeptide of SEQ ID NO: 3 and in another domain a polypeptide of SEQ ID NO: 4, e.g. a chimeric monoclonal antibody. 10 When the antigen binding site comprises both the first and second domains or a polypeptide of SEQ ID NO: 1 or SEQ ID NO:3, respectively, and a polypeptide of SEQ ID NO: 2 or of SEQ ID NO:4, respectively, these may be located on the same polypeptide, or, preferably each domain may be on a different chain, e.g. the first domain being part of an heavy chain, e.g. immunoglobulin heavy chain, or fragment thereof and the second domain being part of a 15 light chain, e.g. an immunoglobulin light chain or fragment thereof. We have further found that a CD45RO/RB binding molecule according to the present invention is a CD45RO/RB binding molecule in mammalian, e.g. human, body environment. A CD45RO/RB binding molecule according to the present invention can thus be designated 20 as a monoclonal antibody (mAb), wherein the binding activity is determined mainly by the CDR regions as described above, e.g. said CDR regions being associated with other molecules without binding specifity, such as framework, e.g. constant regions, which are substantially of human orig in. 25 In another aspect the present invention provides a CD45RO/RB binding molecule which is not the monoclonal antibody "A6" as described by Aversa et al., Cellular Immunology 158, 314-328 (1994). In another aspect the present invention provides a CD45RO/RB binding molecule according 30 to the present invention which is a chimeric, a humanised or a fully human monoclonal antibody. Examples of a CD45RO/RB binding molecules include chimeric or humanised antibodies e.g. derived from antibodies as produced by B-cells or hybridomas and or any fragment -9 thereof, e.g. F(ab')2 and Fab fragments, as well as single chain or single domain antibodies. A single chain antibody consists of the variable regions of antibody heavy and light chains covalently bound by a peptide linker, usually consisting of from 10 to 30 amino acids, preferably from 15 to 25 amino acids. Therefore, such a structure does not include the 5 constant part of the heavy and light chains and it is believed that the small peptide spacer should be less antigenic than a whole constant part. By a chimeric antibody is meant an antibody in which the constant regions of heavy and light chains or both are of human origin while the variable domains of both heavy and light chains are of non-human (e.g. murine) origin. By a humanised antibody Is meant an antibody in which the hypervariable regions 10 (CDRs) are of non-human (e.g. murine) origin while all or substantially all the other part, e.g. the constant regions and the highly conserved parts of the variable regions are of human origins. A humanised antibody may however retain a few amino acids of the murine sequence in the parts of the variable regions adjacent to the hypervariable regions. 15 Hypervariable regions, i.e. CDR's according to the present invention may be associated with any kind of framework regions, e.g. constant parts of the light and heavy chains, of human origin. Suitable framework regions are e.g. described in "Sequences of proteins of immunological interest", Kabat, E.A. et al, US department of health and human services, Public health service, National Institute of health. Preferably the constant part of a human 20 heavy chain may be of the IgG I type, including subtypes, preferably the constant part of a human light chain may be of the K or X type, more preferably of the K type. A preferred constant part of a heavy chain is a polypeptide of SEQ ID NO: 4 (without the CDR1', CDR2' and CDR3' sequence parts which are specified above) and a preferred constant part of a light chain is a polypeptide of SEQ ID NO: 3 (without the CDR1, CDR2 and CDR3 sequence 25 parts which are specified above). We also have found a humanised antibody comprising a light chain variable region of amino acid SEQ ID NO:7 or of amino acid SEQ ID NO:8, which comprises CDR1', CDR2' and CDR3 according to the present invention and a heavy chain variable region of SEQ:ID NO:9 30 or of SEQ:ID NO:10, which comprises CDR1, CDR2 and CDR3 according to the present invention.

-10 In another aspect the present invention provides a humanised antibody comprising a polypeptide of SEQ ID NO:9 or of SEQ ID NO:10 and a polypeptide of SEQ ID NO:7 or of SEQ ID NO:8. 5 In another aspect the present invention provides a humanised antibody comprising - a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:7, - a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:8, - a polypeptide of SEQ ID NO:10 and a polypeptide of SEQ ID NO:7, or - a polypeptide of SEQ ID NO:10 and a polypeptide of SEQ ID NO:8. 10 A polypeptide according to the present invention, e.g. of a herein specified sequence, e.g. of CDR1, CDR2, CDR3, CDR1', CDR2', CDR3, or of a SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10 includes direct equivalents of said (poly)peptide (sequence); e.g. including a functional derivative of 15 said polypeptide. Said functional derivative may include covalent modifications of a specified sequence, and/or said functional derivative may Include amino acid sequence variants of a specified sequence. "Polypeptide". if not otherwise specified herein, includes any peptide or protein comprising 20 amino acids joined to each other by peptide bonds, having an amino acid sequence starting at the N-terminal extremity and ending at the C-terminal extremity. Preferably the polypeptide of the present invention is a monoclonal antibody, more preferred is a chimeric (V-grafted) or humanised (CDR-grafted) monoclonal antibody. The humanised (CDR grafted) monoclonal antibody may or may not include further mutations introduced into the 25 framework (FR) sequences of the acceptor antibody. A functional derivative of a polypeptide as used herein includes a molecule having a qualitative biological activity in common with a polypeptide to the present invention, i.e. having the ability to bind to CD45RO and CD45RB. A functional derivative includes 30 fragments and peptide analogs of a polpypeptide according to the present invention. Fragments comprise regions within the sequence of a polypeptide according to the present invention, e.g. of a specified sequence. The term "derivative" is used to define amino acid sequence variants, and covalent modifications of a polypeptide according to the present Invention. e.g. of a specified sequence. The functional derivatives of a polypeptide according - 11 to the present invention, e.g. of a specified sequence, preferably have at least about 65%, more preferably at least about 75%, even more preferably at least about 85%, most preferably at least about 95% overall sequence homolog y with the amino acid sequence of a polypeptide according to the present invention, e.g. of a specified sequence, and 5 substantially retain the ability to bind to CD45RO and CD45RB. The term "covalent modification" includes modifications of a polypeptide according to the present invention, e.g. of a specified sequence; or a fragment thereof with an organic proteinaceous or non-proteinaceous derivatizing agent, fusions to heterologous polypeptide 10 sequences, and post-translational modifications. Covalent modified polypeptides, e.g. of a specified sequence, still have the ability bind to CD45RO and CD45RB by crosslinking. Covalent modifications are traditionally introduced by reacting targeted amino acid residues with an organic derivatizing agent that is capable of reacting with selected sides or terminal residues, or by harnessing mechanisms of post-translational modifications that function in 15 selected recombinant host cells. Certain post-translational modifications are the result of the action of recombinant host cells on the expressed polypeptide. Glutaminyl and asparaginyl residues are frequently post-translationally deamidated to the corresponding glutamyl and aspartyl residues. Alternatively, these residues are deaminated under mildly acidic conditions. Other post-translational modifications include hydroxylation of proline and lysine, 20 phosphorylation of hydroxyl groups of seryl, tyrosine or threonyl residues, methylation of the a-amino groups of lysine, arginine, and histidine side chains, see e.g. T. E. Creighton, Proteins: Structure and Molecular Properties, W. H. Freeman & Co., San Francisco, pp. 79 86 (1983). Covalent modifications e.g. include fusion proteins comprising a polypeptide according to the present invention, e.g. of a specified sequence and their amino acid 25 sequence variants, such as immunoadhesins, and N-terminal fusions to heterologous signal sequences. "Homology" with respect to a native polypeptide and its functional derivative is defined herein as the percentage of amino acid residues in the candidate sequence that are identical with 30 the residues of a corresponding native polypeptide, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology, and not considering any conservative substitutions as part of the sequence identity. Neither N- or C terminal extensions nor insertions shall be construed as reducing identity or homology. Methods and computer programs for the alignment are well known.

- 12 "Amino acid(s)" refer to all naturally occurring L-a-amino acids, e.g. and including D-amino acids. The amino acids are identified by either the well known single-letter or three-letter designations. 5 The term "amino acid sequence variant" refers to molecules with some differences in their amino acid sequences as compared to a polypeptide according to the present invention, e.g. of a specified sequence. Amino acid sequence variants of a polypeptide according to the present invention, e.g. of a specified sequence, still have the ability to bind to CD45RO and CD45RB. Substitutional variants are those that have at least one amino acid residue 10 removed and a different amino acid inserted in its place at the same position in a polypeptide according to the present Invention, e.g. of a specified sequence. These substitutions may be single, where only one amino acid in the molecule has been substituted, or they may be multiple, where two or more amino acids have been substituted In the same molecule. Insertional variants are those with one or more amino acids inserted immediately adjacent to 15 an amino acid at a particular position in a polypeptide according to the present invention, e.g. of a specified sequence. Immediately adjacent to an amino acid means connected to either the a-carboxy or a-amino functional group of the amino acid. Deletional variants are those with one or more amino acids in a polypeptide according to the present invention, e.g. of a specified sequence, removed. Ordinarily, deletional variants will have one or two amino acids 20 deleted in a particular region of the molecule. We also have found the polynucleotide sequences of - GGCCAGTCAGAACATTGGCACAAGCATACAGTG, encoding the amino acid sequence of CDR1, 25 - TTCTTCTGAGTCTATCTCTGG; encoding the amino acid sequence of CDR 2, - ACAAAGTAATACCTGGCCATTCACGTT encoding the amino acid sequence of CDR 3, - TTATATTATCCACTG, encoding the amino acid sequence of CDR1', - TTTTAATCCTTACAATCATGGTACTAAGTACAATGAGAAGTTCAAAGGCAG encoding the amino acid sequence of CDR2', 30 AGGACCCTATGCCTGGTTTGACACCTG encoding the amino acid sequence of CDR3', - SEQ ID NO:5 encoding a polypeptide of SEQ ID NO: 1, I.e. the variable region of a light chain of an mAb according to the present Invention; - SEQ ID NO:6 encoding a polypeptide of SEQ ID NO:2, I.e. the variable region of the heavy chain of an mAb according to the present invention; - 13 - SEQ ID NO:11 encoding a polypeptide of SEQ ID NO:9. i.e. a heavy chain variable region including CDR1, CDR2 and CDR3 according to the present invention; - SEQ ID NO:12 encoding a polypeptide of SEQ ID NO:10, i.e. a heavy chain variable region including CDR1, CDR2 and CDR3 according to the present invention; 5 - SEQ ID NO:13 encoding a polypeptide of SEQ ID NO:7, i.e. a light chain variable region including CDR1', CDR2' and CDR3' according to the present invention; and - SEQ ID NO:14 encoding a polypeptide of SEQ ID NO:8, i.e. a light chain variable region including CDR1', CDR2' and CDR3' according to the present invention. 10 In another aspect the present invention provides isolated polynucleotides comprising polynucleotides encoding a CD45RO/RB binding molecule, e.g. encoding the amino acid sequence of CDR1, CDR2 and CDR3 according to the present invention and/or, preferably and, polynucletides encoding the amino acid sequence of CDR1', CDR2' and CDR3' according to the present invention; and 15 Polynucleotides comprising a polynucleotide of SEQ ID NO: 5 and/or, preferably and, a polynucleotide of SEQ ID NO: 6; and Polynucleotides comprising polynucleotides encoding a polypeptide of SEQ ID NO:7 or SEQ ID NO:8 and a polypeptide of SEQ ID NO:9 or SEQ ID NO:10; e.g. encoding - a polypeptide of SEQ ID NO:7 and a polypeptide of SEQ ID NO:9, 20 - a polypeptide of SEQ ID NO:7 and a polypeptide of SEQ ID NO:10, - a polypeptide of SEQ ID NO:8 and a polypeptide of SEQ ID NO:9, or - a polypeptide of SEQ ID NO:8 and a polypeptide of SEQ ID NO:10; and Polynucleotides comprising a polynucleotide of SEQ ID NO:11 or of SEQ ID NO:12 and a polynucleotide of SEQ ID NO:13 or a polynucleotide of SEQ ID NO:14, preferably 25 comprising - a polynucleotide of SEQ ID NO:11 and a polynucleotide of SEQ ID NO:13, - a polynucleotide of SEQ ID NO:1 I and a polynucleotide of SEQ ID NO:14, - a polynucleotide of SEQ ID NO:12 and a polynucleotide of SEQ ID NO:13, or - a polynucleotide of SEQ ID NO:12 and a polynucleotide of SEQ ID NO:14. 30 "Polynucleotide", if not otherwise specified herein, includes any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA, or modified RNA or DNA, including without limitation single and double stranded RNA, and RNA that is a mixture of single- and double-stranded regions.

-14 A polynucleotide according to the present invention, e.g. a polynucleotide encoding the amino acid sequence CDR1, CDR2, CDR3, CDR1', CDR2', CDR3', or of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ 5 ID NO:10, respectively, such as a polynucleotide of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:1 1, SEQ ID NO:12, SEQ ID NO:13 or SEQ ID NO:14, respectively, includes allelic variants thereof and/or their complements; e.g. including a polynucleotide that hybridizes to the nucleotide sequence of SEQ ID NO: 5, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13 or SEQ ID NO:14, respectively; e.g. encoding a polypeptide having at least 10 80% identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, e.g. Including a functional derivative of said polypeptide, e.g. said functional derivative having at least 65% homology with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, e.g. said functional derivative including 15 covalent modifications of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, e.g. said functional derivative including amino acid sequence variants of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively; e.g. a SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:1 1, SEQ ID NO:12, SEQ ID 20 NO:13 or SEQ ID NO:14, respectively includes a sequence, which as a result of the redundancy (degeneracy) of the genetic code, also encodes a polypeptide of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, or encodes a polypeptide with an amino acid sequence which has at least 80% identity with the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ 25 ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively. A CD45RO/RB binding molecule, e.g. which is a chimeric or humanised antibody, may be produced by recombinant DNA techniques. Thus, one or more DNA molecules encoding the 30 CD45RO/RB may be constructed, placed under appropriate control sequences and transferred into a suitable host (organism) for expression by an appropriate vector. In another aspect the present invention provides a polynucleotide which encodes a single, heavy and/or a light chain of a CD45RO/RB binding molecule according to the present -15 Invention; and the use of a polynucleotide according to the present invention for the production of a CD45RO/RB binding molecule according to the present invention by recombinant means. 5 A CD45RO/RB binding molecule may be obtained according, e.g. analogously, to a method as conventional together with the information provided herein, e.g. with the knowledge of the amino acid sequence of the hypervarlable or variable regions and the polynucleotide sequences encoding these regions. A method for constructing a variable domain gene is e.g. described in EP 239 400 and may be briefly summarized as follows: A gene encoding a 10 variable region of a mAb of whatever specificity may be cloned. The DNA segments encoding the framework and hypervarlable regions are determined and the DNA segments encoding the hypervariable regions are removed. Double stranded synthetic CDR cassettes are prepared by DNA synthesis according to the CDR and CDR' sequences as specified herein. These cassettes are provided with sticky ends so that they can be ligated at junctions 15 of a desired framework of human origin. Polynucleotides encoding single chain antibodies may also be prepared according to, e.g. analogously, to a method as conventional. A polynucleotide according to the present invention thus prepared may be conveniently transferred into an appropriate expression vector. 20 Appropriate cell lines may be found according, e.g. analogously, to a method as conventional. Expression vectors, e.g. comprising suitable promotor(s) and genes encoding heavy and light chain constant parts are known e.g. and are commercially available. Appropriate hosts are known or may be found according, e.g. analogously, to a method as conventional and include cell culture or transgenic animals. 25 In another aspect the present invention provides an expression vector comprising a polynucleotide encoding a CD45RO/RB binding molecule according to the present invention, e.g. of sequence SEQ ID NO:15, SEQ ID NO:16,. SEQ ID NO:17 or SEQ ID NO:18. 30 In another aspect the present invention provides - An expression system comprising a polynucleotide according to the present invention wherein said expression system or part thereof is capable of producing a CD45RO/RB binding molecule according to the present invention, when said expression system or part thereof is present in a compatible host cell; - 16 and - An isolated host cell which comprises an expression system as defined above. We have further found that a CD45RO/RB binding molecule according to the present 5 invention inhibit primary alloimmune responses In a dose-dependent fashion as determined by in vitro MLR. The results indicate that the cells which had been alloactivated in the presence of a CD45RO/RB binding molecule according to the present invention are impaired in their responses to alloantigen. This confirms the indication that a CD45RO/RB binding molecule according to the present invention can act directly on the effector alloreactive T 10 cells and modulate their function. In addition, the functional properties of T cells derived from the primary MLR were further studied in restimulation experiments In secondary MLR, using specific stimulator cells or third-party stimulators to assess the specificity of the observed functional effects. We have found that the cells derived from primary MLRs in which a CD45RO/RB binding molecule according to the present Invention is present, were impaired 15 in their ability to respond to subsequent optimal stimulation with specific stimulator cells, although there was no antibody added to the secondary cultures. The specificity of the inhibition was demonstrated by the ability of cells treated with a CD45RO/RB binding molecule according to the present invention to respond normally to stimulator cells from unrelated third-party donors. Restimulation experiments using T cells derived from primary 20 MLR cultures thus indicate that the cells which had been alloactivated a CD45RO/RB binding molecule according to the present invention are hyporesponsive, I.e. tolerant, to the original alloantigen. Furthermore we have found that cell proliferation in cells pre-treated with a CD45RO/RB 25 binding molecule according to the present invention could be rescued by exogenous IL-2. This Indicates that treatment of alloreactive T cells with a CD45RO/RB binding molecule according to the present invention induces a state of tolerance. Indeed, the reduced proliferative responses observed in cells treated with a CD45RO/RB binding molecule according to the present invention, was due to impairement of T cell function, and these 30 cells were able to respond to exogenous IL-2, indicating that these cells are in an anergic, true unresponsive state. The specificity of this response was shown by the ability of cells treated with a CD45RO/RB binding molecule according to the present Invention to proliferate normally to unrelated donor cells to the level of the control treated cells.

- 17 In addition experiments indicate that the binding of a CD45RO/RB binding molecule according to the present invention to CD45RO and CD45RB may inhibit the memory responses of peripheral blood mononuclear cells (PBMC) from immunized donors to specific recall antigen. Binding of a CD45RO/RB binding molecule according to the present invention 5 to CD45RO and CD45RB thus is also effective in inhibiting memory responses to soluble Ag. The ability of a CD45RO/RB binding molecule according to the present invention to inhibit recall responses to tetanus in PBMC from immunized donors Indicate that the a CD45RO/RB binding molecule according to the present invention is able to target and modulate the activation of memory T cells. E.g. these data indicate that a CD45RO/RB binding molecule 10 according to the present invention in addition to recognizing alloreactive and activated T cells is able to modulate their function, resulting in induction of T cell anergy. This property may be Important in treatment of ongoing immune responses to autoantigens and allergens and possibly to alloantigens as seen in autoimmune diseases, allergy and chronic rejection, and diseases, such as psoriasis, inflammatory bowel disease, where memory responses play a 15 role in the maintenance of disease state. It is believed to be an Important feature in a disease situation, such as in autoimmune diseases in which memory responses to autoantig ens may play a major role for the disease maintenance. We have also found that a CD45RO/RB binding molecule according to the present invention 20 may modulate T cell proliferative responses in a mixed lymphocyte response (MLR) in vivo, i.e. a CD45RO/RB binding molecule according to the present invention was found to have corresponding inhibitory properties in vivo testing. A CD45RO/RB binding molecule according to the present invention may thus have 25 immunosuppressive and tolerogenic properties and may be useful for in vivo and ex-vivo tolerance induction to alloantigens, autoantigens, allergens and bacterial flora antigens, e.g. a CD45RO/RB binding molecule according to the present invention may be useful in the treatment and prophylaxis of diseases e.g. including autoimmune diseases, such as, but not limited to, rheumatoid arthritis, autoimmune thyroditis, Graves disease, type I and type I 30 diabetes, multiple sclerosis, systemic lupus erythematosus, Sj6gren syndrome, scleroderma, autoimmune gastritis, glomerulonephritis, transplant rejection, e.g. organ and tissue allograft and xenograft rejection, graft versus host disease (GVHD), and also psoriasis, inflammatory bowel disease and allergies.

- 18 In another aspect the present Invention provides the use of a CD45RO/RB binding molecule according to the present Invention as a pharmaceutical, e.g. In the treatment and prophylaxis of autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies. 5 In another aspect the present invention provides a CD45RO/RB binding molecule according to the present invention for the production of a medicament in the treatment and prophylaxis of diseases associated with autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies. 10 In another aspect the present invention provides a pharmaceutical composition comprising a CD45RO/RB binding molecule according to the present invention in association with at least one pharmaceutically acceptable carrier or diluent. 15 A pharmaceutical composition may comprise further, e.g. active, ingredients, e.g. other immunomodulatory antibodies such as, but not confined to anti-ICOS, anti-CD154, anti CD134L or recombinant proteins such as, but not confined to rCTLA-4 (CD152), rOX40 (CD134), or immunomodulatory com pounds such as, but not confined to cyclosporin A, FTY720, RAD, rapamycin, FK506, 15-deoxyspergualin, steroids. 20 In another aspect the present invention provides a method of treatment and/or prophylaxis of diseases associated with autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies comprising administering to a subject In need of such treatment and/or prophylaxis an effective amount of a CD45RO/RB binding molecule according to the 25 present invention, e.g. in the form of a pharmaceutical composition according to the present invention. Autoimune diseases to be treated with binding molecule of the present Invention further include, but are not limited to, rheumatoid arthritis, autoimmune thyroditis, Graves disease, 30 type I and type I diabetes, multiple sclerosis, systemic lupus erythematosus, Sj6gren syndrome, scleroderma, autoimmune gastritis, glomerulonephritls; transplant rejection, e.g. organ and tissue allograft and xenograft rejection and graft-versus-host disease (GVHD).

-19 EXAMPLES The invention will be more fully understood by reference to the following examples. They 5 should not, however, be construed as limiting the scope of the invention. In the following examples all temperatures are in degree Celsius. The "candidate mAb" or "chimeric antibody" is a CD45RO/RB binding molecule according to the present invention comprising light chain of SEQ ID NO:3 and heavy chain of SEQ ID 10 NO:4. The following abbreviations are used: ELISA enzyme linked Immuno-sorbant assay FACS fluorescence activated cell sorting 15 FITC fluorescein isothiocyanate FBS foetal bovine serum GVHD graft-vs-host disease HCMV human cytomegalovirus promoter IgE immunoglobulin isotype E 20 IgG immunoglobulin isotype G PBS phosphate-buffered saline PCR polymerase chain reaction xGVHD xeno-graft-vs-host disease 25 -20 Example 1: Primary mixed lymphocyte response (MLR) Cells 5 Blood samples are obtained from healthy human donors. Peripheral blood mononuclear cells (PBMC) are isolated by centrifugation over Ficoll-Hypaque (Pharmacia LKB) from leukocytes from whole peripheral blood, leukopheres is or buffy coats with known blood type, but unknown HLA type. In some MLR experiments, PBMC are directly used as the stimulator cells after the Irradiation at 40 Gy. In the other experiments, T cells were depleted from 10 PBMC by using CD2 or CD3 Dynabeads (Dynal, Oslo, Norway). Beads and contaminating cells are removed by magnetic field. T cell-depleted PBMC are used as simulator cells after the irradiation. PBMC, CD3* T cells or CD4' T cells are used as the responder cells in MLR. Cells are prepared from different donors to stimulator cells. CD3' T cells are purified by negative 15 selection using antl-CD16 mAb (Zymed, CA), goat anti-mouse IgG Dynabeads, anti-CD14 Dynabeads, CD19 Dynabeads. In addition ant i-CD8 Dynabeads are used to purify CD4' T cells. The cells obtained are analyzed by FACScan or FACSCalibur (Becton Dickinson & Co., CA) and the purity of the cells obtained was >75%. Cells are suspended in RPM11640 medium, supplemented with 10 % heat-inactivated FBS, penicillin, streptomycin and L 20 glutamine. Reagents The chimeric anti-CD45RO/RB mAb "candidate mAb" and an isotype matched control chimeric antibody is also generated. Mouse (Human) control IgG, antibody specific for KLH 25 (keyhole limpet hemocyanin) or recombinant human IL-10 is purchased from BD Pharmingen (San Diego, CA). Anti-human CD154 mAb 5c8 is according to Lederman et al 1992. Primary Mixed lymphocyte response (MLR) 30 Aliquots of 1 x 10o PBMC or 5 X 10 4 of CD3' or CD4' cells are mixed with 1 x 105 irradiated PBMC or 5 x 104 T cells-depleted Irradiated (50 Gy) PBMC in the each well of 96-well culture plates (Costar, Cambridge, MA) In the presence of the indicated mAb or absence of Ab. In some experiments, F(ab') 2 fragment of goat anti-mouse Ig or goat anti-human Ig specific for -21 Fc portion (Jackson ImmunoResearch, West Grove, PA) is added at 10 pg/ml in addition to the candidate mAb To ensure optimal in vitro cross-linking of the target CD45 molecules. The mixed cells are cultured for 4 or 5 days at 37 0 C in 5% CO 2 and proliferation is determined by pulsing the cells with 3 H-thymidine for the last 16 - 20 hours of culture. 5 Other experiments are similar to those described above, but with the following exceptions: 1) Medium used is EX-VIVO (Bio-Whittaker) containing 10% FBS and 1% human plasma; 2) Anti-mouse total IgG (5 pg/ml) is used as secondary cross-linking step; 3) Irradiation of stimulator cells is 60 Gy. Primary MLR is performed in the presence of the "candidate mAb" or control chimeric IgG, 10 (10 pg/ml) both with a second step reagent, F(ab') 2 fragment of goat anti-human Ig specific for Fc portion (10 pg/ml). Percentage inhibition by the "candidate mAb" is calculated in comparison with the cell proliferation in the presence of control IgG 1 . Results are shown in TABLE 1 below: TABLE 1 15 Inhibition of primary MLR by 10 pg/ml of a candidate mAb according to the present invention Responder Stimulator (Irr. PBMC) % of Inhibition #211 CD4 #219 CD3 63.51 #220 CD4 #219 CD3 depl. 63.07 #227 CD4 #220 CD3 depl. 65.96 #229 CD4 #219 CD3 depl. 50.76 Average+ SD 60.83 +6.83 * * Significantly different from control value (P<0.001) A candidate mAb according to the present invention inhibits primary MLR as can be seen from TABLE 1. The average inhibitory effect is 60.83 + 6.83 % in four different donors 20 derived CD4' T cells and statistically significant. The inhibition of primary MLR by the "candidate mAb" is shown to be dose-dependent in the range of 0.001 and 10 pg/mI of the "candidate mAb" as shown in Figure 1. The IC50 for the inhibition of primary MLR by a "candidate mAb" is determined from the results of three separate MLR experiments using one donor PBMC as responder cells. Thus, 25 responder CD4' T cells from Donor #229 and #219 and irradiated PBMC depleted of T cells as stimulators are mixed in the presence of a "candidate mAb" or control chimeric Ab with 10 pg/mI of F(ab') 2 fragment of goat anti-human Ig. Experiments are repeated 3 times and percentage of proliferation in the presence of a "candidate mAb" is calculated in comparison - 22 with the T cell proliferation in the presence of control Ab. ICso value is determined using Origin (V. 6.0@). The cellular activity ICso value is calculated to be 0.87 + 0.35 nM (0.13 + 0.052 pg/ml). 5 Example 2: Secondary MLR In order to assess whether a "candidate mAb" induces unresponsiveness of CD4' T cells to specific alloantigens, secondary MLR is performed in the absence of any antibodies after the primary MLC. CD4' T cells are cultured with irradiated allogeneic stimulator cells (T cells 10 depleted PBMC) in the presence of the indicated antibody in 96-well culture plates for 10 days (primary MLC). Then, cells are collected, layered on a Ficoll-Hypaque gradient to remove dead cells, washed twice with RPMI, and restimulated with the same stimulator, 3" party stimulator cells or IL-2 (50 U/ml). The cells are cultured for 3 days and the proliferative response is determined by pulsing the cells with 3 H-thymidine for the last 16 - 20 hours of 15 culture. Specifically, CD4' T cells are cultured with irradiated allogeneic stimulator cells (T cells depleted PBMC taken from other donors) in the presence of 10 pig/ml of the "candidate mAb", control IgG1 chimeric Ab and F(ab') 2 fragment of goat anti-human ig. Primary MLR proliferation is determined on day 5. For secondary MLR, the responder and stimulator cells 20 are cultured for 10 days in the presence of the "candidate mAb", then the cells are harvested, washed twice In RPM11640 and restimulated with specific stimulator, third-party stimulators or IL-2 (50 U/ml) in the absence of any Ab. Cell proliferation is determined on day 3. Results set out in TABLE 2: TABLE 2 Responder CD4+ T cells Donor # % Inhibition of 2 rY MLR #211 49.90* #220 59.33* #227 58.68* 25 * Significantly different from control value (p=cO.001 determined by t-test, SigmaStat V.2.03). # p=<0.046 In order to test whether the impaired proliferation Is due to unresponsivess as a consequence of the treatment with a "candidate mAb", the cells derived from primary MLR 30 are cultured In the presence of IL-2 (50 U/ml). Addition of IL-2 results in the rescue of -23 proliferative responses of the T cells which had been treated with a "candidate mAb" in primary MLR, to levels similar to those observed in the presence of IgG 1 control Ab. These data indicate that the impaired secondary response in T cells treated with a "candidate mAb" is due to to functional alteration of the responder T cells which become unresponsive to the 5 specific stimulator cells. Percentage inhibition is calculated according to the following formula: c.p.m. with control Ab - c.p.m. with "candidate mAb x 100 c.p.m. with control Ab 10 Statistical analysis is performed using SigmaStat (Vers. 2.03). The data is analyzed by two-way ANOVA followed by Dunnett method. In all test procedures probabilities <0.05 are considered as significant. In some experiments t-test is used (SigmaStat V.2.03). 15 Example 3: In vivo survival studies In SCID-mice Engraftment of hu-PBL in SCID mice Human peripheral blood mononuclear cells (PBMC) are injected intraperitoneally into SCID 20 mice C.B 17 /GbmsTac-Prkdc" Lystb mice (Taconic, Germantown, NY) in an amount sufficient to induce a lethal xenogeneic graft-versus-host disease (xGvHD) in >90% of the mice within 4 weeks after cell transfer. Such treated SCID mice are hereinafter designated as hu-PBL-SCID mice 25 Mab-treatment of hu-PBL-SCID mice Hu-PBL-SCID mice are treated with a "candidate mAb" or mouse or chimeric isotype matched mAb controls at day 0, immediately after PBMC injection, at day 3, day 7 and at weekly intervals thereafter. Mabs are delivered subcutaneously in 100 pl PBS at a final concentration of 5 mg/kg body weight. The treatment was stopped when all control mice 30 were dead. Evaluation of treatment results The main criterion to assess the efficacy of a "candidate mAb" In this study was the survival of the hu-PBL-SCID mice. The significance of the results is evaluated by the statistical -24 method of survival analysis using the Log-rank test (Mantel method) with the help of the Systat v9.01 software. The method of survival analysis is a non-parametric test, which not only consider whether a particular mouse is still alive but also whether if it was sacrificed for reasons irrelevant to the treatment/disease such as the requirement of perform in vitro 5 analysis with its organs/cells. Biopsies of liver, lung, kidney and spleen are obtained from dead mice for further evaluation. In addition, hu-PBL-SCID mice are weighed at the beginning (before cell transfer) and throughout (every two days) the experiment as an indirect estimation of their health status. Linear regression lines were generated using the body weight versus days post-PBMC transfer values obtained from each mouse and 10 subsequently, their slopes (control versus anti-CD45 treated mice) were compared using the non-parametric Mann-Whitney test. Results All hu-PBL-SCID mice treated with mouse mAb controls had infiltrated human leukocytes in 15 the lung, liver and spleen and died (4/4) within ca. 2 to 3 weeks after cell transfer. Death is a likely consequence of xGvHD. Control mAb-treated mice furthermore lost weight in a linear manner, ca. 10% and more within 3 weeks. All hu-PBL-SCID mice treated with a "candidate mAb" survived (4/4) without any apparent sign of disease more than 4 weeks, even although "candidate mAb"-treatment was stopped 20 after 3 weeks. "Candidate mAb"-treated mice increased weight in a linear manner, up to ca. 5% within 4 weeks. Example 4: Expression of antibodies of the invention 25 Expression of humanised antibody comprising a SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10 Expression vectors according to the plasmid map shown in Figures 2 to 5 are constructed, comprising the corresponding nucleotides encoding the amino acid sequence of humanised 30 light chain variable region humVi (SEQ ID NO:7), humanised light chain variable region humV2 (SEQ ID NO:8), humanised heavy chain variable region VHE (SEQ ID NO:9), or humanised heavy chain variable region VHQ (SEQ ID NO:10), respectively. These expression vectors have the DNA (nucleotide) sequences SEQ ID NO 15, SEQ ID NO 16, SEQ ID NO 17, or SEQ ID NO 18, respectively.

- 25 Construction of humanised antibody heavy and light chain express ion vectors Human kappa light chain expression vectors for versions VLh and VLm In order to construct the final expression vector encoding for the complete humanised light 5 chain of human kappa isotype, DNA fragments encoding the complete light chain variable regions (VLh and VLm) were excised from the VLh and VLm containing PCR-Script cloning vectors (Stratagene) (VLm region) using HindIll and Bglll. The gel-purified fragments were then subcloned into the HindlIl and BamHI sites of C21-HCMV Kappa expression vector which was created during construction of the humanised anti-IgE antibody TESC-21 10 (Kolbinger et al 1993) and which originally received from M. Bendig (MRC Collaborative Centre, London, UK) (Maeda et al. 1991). The ligation products were purified by phenol/chloroform extraction, and electroporated into electrocoporation-competent Epicurlan Coli@ XL1-Blue strain (Cat. N' #200228, Stratagene). After plating on LB/amp agar plates overnight at 370C, each 12 colonies were picked to prepare plasmid DNA from a 3 ml culture 15 using the BioRobot 9600 (Qiagen). This yielded the light chain expression vectors for the humanised antibody versions VLh and VLm, respectively, as further described in the Figures. Human gamma-1 heavy chain expression vectors for VHQ 20 For the construction of the VHQ expression vector, a step-wise approach was taken. First, the complete variable region of VHQ was assembled by PCR according to the methology as described in Kolbinger et al 1993 (Protein Eng. 1993 Nov; 6(8):971-80) and subcloned into the C21-HCMV-gamma-1 expression from which the C21 Insert had been removed using the same enzymes. A Hindlll/BamHl fragment of PCRScript clone VHQ containing the complete 25 variable region was then subcloned into expression vector C21-HCMV-gamma-1 cleaved with the same enzymes. This yielded the final expression vector for the humanised antibody version VHQ. Human gamma-1 heavy chain expression vectors for VHE 30 The construction of the final VHE expression vector encoding for the complete humanised heavy chain of human gamma-1 Isotype was achieved by directly ligating a Hindill and -26 BamHI restricted PCR fragment encoding the variable region into the Hindill and BamHI sites of C21-HCMV gamma-1 expression vector which was created during construction of the humanised anti-IgE antibody TESC-21 (Kolbinger et al 1993) and which was also originally received from M. Bendig (MRC Collaborative Centre, London, UK) (Maeda et al. 5 1991). Transient expression in COS cells The following transfection protocol is adapted for adherent COS cells in 150 mm cell culture 10 dishes, using SuperFect Tm Transfection Reagent (Cat. N'301305, Qiagen). The four different expression vectors described above are used for transient transfection of cells. For expression of humanised antibody, each of two clones containing heavy chain inserts (VHE or VHQ, respectively) are co-transfected into cells with each of the two clones encoding for the light chains (humV1 or humV2, respectively), in total 4 different combinations of heavy 15 and light chain expression vectors (VHE/humV1, VHE/humV2, VHQ/humV1 and VHQ/humV2). Before transfection, the plasmids are linearized with the restriction endonuclease Pvul which cleaves in the region encoding the resistance gene for ampicillin. The day before transfection, 4 x 10' COS cells in 30 ml of fresh culture medium are seeded in 150 mm cell culture dishes. Seeding at this cell density generally yielded 80% confluency 20 after 24 hours. On the day of transfection, four different combinations of linearized heavy and light-chain DNA expression vectors (15 pg each) are diluted in a total volume of 900 pl of fresh medium without serum and antibiotics. 180 pl of SuperFect Transfection Reagent is then mixed thoroughly with the DNA solution. The DNA mixture is incubated for 10 min at room temperature to allow complex formation. While complex formation takes place, the 25 growth medium is removed from COS cell cultures, and cells are washed once with PBS. 9 ml of fresh culture medium (containing 10% FBS and antibiotics) are then added to each reaction tube containing the transfection complexes and well mixed. The final preparation is immediately transferred to each of 4 cultures to be transfected and gently mixed. Cell cultures are then incubated with the DNA complexes for 3 hours at 37*C and 5% C02. After 30 incubation, the medium containing transfection complexes Is removed and replaced with 30 ml of fresh culture medium. At 48 hr post transfection, the culture supematants are harvested. Concentration of culture supematants - 27 For ELISA and FACS analysis, the culture supematants collected from COS cells transfected with heavy- and light- chain plasmids are concentrated as follows. 10 ml of each supernatant are added to Centriprep YM-50 Centrifugal Filter Devices (Cat. N' 4310, Millipore) as described by the manufacturer. The Centriprep filters are centrifuged for 10 min 5 at 3000 rpm at room temperature. The centrifugation step is then repeated again with the remaining 20 ml of supernatant using only 5 min of centrifugation and supervising the concentration evolution. The intermediate 500 pl of concentrated supernatant is recovered, transferred to new Microcon Centrifugal Filter Devices (Cat. N' 42412, Microcon) and further concentrated following the manufacturer's protocol. The concentrated supematants are 10 centrifuged four times for 24 min at 3000 rpm at room temperature, one time for 10 min at 6000 rpm and then, three times for 5 min, always supervising the concentration evolution. The final volume of concentrated conditioned medium achieved is 100-120 pi corresponding to a 250 to 300-fold concentration of original culture medium and Is stored at 4 0 C until use. For comparison and control, culture medium from untransfected cells is similarly 15 concentrated, using the same centrifugation protocol described above. Example 5: Determination of recombinant human IgG expression by ELISA To determine IgG concentrations of recombinant human antibody expressed in the culture 20 supematants, a sandwich ELISA protocol has been developed and optimized using human IgG as standard. Flat bottom 96-well microtiter plates (Cat. N' 4-39454, Nunc Immunoplate Maxisorp) are coated overnight at 4'C with 100 pl of goat anti-human IgG (whole molecule, Cat. N* 11011, SIGMA) at the final concentration of 0.5 pg/ml in PBS. Wells are then washed 3 times with washing buffer (PBS containing 0.05% Tween 20) and blocked for 1.5 hours at 25 37 0 C with blocking buffer (0.5% BSA in PBS). After 3 washing cycles, the antibody samples and the standard human I gG (Cat. No. 14506, SIGMA) are prepared by serial 1. 5-fold dilution in blocking buffer. 100 pl of diluted samples or standard are transfered in duplicate to the coated plate and incubated for I hour at room temperature. After incubation, the plates are washed 3 times with washing buffer and subsequently incubated for 1 hour with 100 pi of 30 horseradish peroxidase-conjugated goat anti-human IgG kappa-light chain (Cat. N' A-7164, SIGMA) diluted at 1/4000 in blocking buffer. Control wells received 100 pl of blocking buffer or concentrated normal culture medium. After washing, the colorimetric quantification of bound peroxidase in the sample and standard wells Is performed, using a TMB Peroxidase EIA Substrate Kit (Cat. N' 172-1067, Bio-Rad) according to the manufacturer's Instructions.

- 28 The peroxidase mixture is added at 100 pi per well and incubated for 30 min at room temperature in the dark. The colorimetric reaction is stopped by addition of 100 pl of I M sulfuric acid and the absorbance in each well is read at 450 nm, using an ELISA plate reader (Model 3350-UV, BioRad). 5 With a correlation coefficient of 0.998 for the IgG standard curve, the following concentrations are determined for the four different culture concentrates (ca. 250-300 fold concentrated): 10 VHE/humV1 supernatant = 8.26 pg/mI VHE/humV2 supernatant = 6.27 pg/ml VHQ/humV1 supernatant = 5.3 pg/mI VHQ/humV2 supernatant = 5.56 pg/mI 15 Example 6: FACS competition analysis (binding affinity) The human T-cell line PEER is chosen as the target cell for FACS analysis because it expressed the CD45 antigen on its cell surface. To analyze the binding affinity of humanised antibody supematants, competition experiments using FITC-Iabeled chimeric antibody as a 20 reference are performed and compared with the inhibition of purified mouse antibody and of chimeric antibody. PEER cell cultures are centrifuged for 10 seconds at 3000 rpm and the medium is removed. Cells are resuspended in FACS buffer (PBS containing 1% FBS and 0.1% sodium azide) and seeded int o 96-well round-bottom microtitter plate at a cell density of 1x105 cells per well. The plate is centrifuged and the supernatant is discarded. For 25 blocking studies, 25 pl of concentrated untransfected medium or isotype matched control antibody (negative controls), unlabeled mouse antibody or chimeric antibody (positive controls) as well as concentrated supernatant containing the various combinations of humanised antibody (samples), is first added In each well at the indicated concentrations in the text. After I hour of incubation at 4*C, PEER cells are washed with 200 pl of FACS 30 buffer by centrifugation. Cells are subsequently incubated for I hour at 40C with chimeric antibody conjugated with FITC In 25 pi of FACS buffer at the final concentration of 20 pg/ml. Cells are washed and resuspended In 300 pl of FACS buffer containing 2 pg/ml propidium iodide which allows gating of viable cells. The cell preparations are analyzed on a flow cytometer (FACSCalibur, Becton Dickinson).

-29 FACS analysis indicates a dose-dependent blockade of fluorochrome-labeled chimeric antibody by the concentrated humanised antibody culture supematants. No dose-dependent blockade of chimeric antibody binding is seen with the isotype matched control antibody, indicating that the blocking effect by the different humanised antibody combinations is 5 epitope specific and that epitope specificity appears to be retained after the humanisation process. Example 7: Biological activities of CD45RB/RO binding molecules 10 In this study, we have addressed whether CD45RB/RO binding chimeric antibody, when present in cultures of polyclonally activated primary human T cells (i) supports the differentiation of T cells with a characteristic Treg phenotype, (ii) prevents or enhances apoptosis following T cell activation, and (iii) affects expression of subset-specific antigens and receptors after restimulation. 15 CD45RB/RO binding chimeric antibody enhances cell death In polyclonally activated T cells Primary T cells (mixture of CD4+ and CD8+ T subsets) were subjected to activation by anti CD3 plus anti-CD28 mAb (200 ng/ml each) in the presence or absence (=control) of 20 CD45RB/RO binding chimeric antibody. Excess antibodies were removed by washing on day 2. 7-amino-actinomycin D (7-AAD) as a DNA-staining dye taken up by apoptotic and necrotic cells was used to measure cell death following activation. The results show that activation of T cells in the presence of CD45RB/RO binding chimeric antibody increased the fraction of 7 AAD positive cells than two-fold on day 2 after activation.. On day 7, the portion of 7-AAD 25 positive cells was again similar in CD45RB/RO binding chimeric antibody-treated and control cultures. CD45RB/RO binding chimeric antibody but not control mAb treated T cells display a T regulatory cell (Treg) phenotype 30 Increased expression of CD25 and the negative regulatory protein CTLA-4 (CD152) is a marker of Treg cells. Functional suppression of primary and secondary T cell responses by CD45RB/RO binding chimeric antibody may be due to the induction of Treg cells. To address this issue, T cells were activated by anti-CD3 + CD28 mAbs and cultured in the presence of CD45RB/RO binding chimeric antibody or anti-LPS control mAb. The time - 30 course of CTLA-4 and CD25 expression reveals marked differences between controls and CD45RB/RO binding chimeric antibody-treated T cells on days 1 and 3 after secondary stimulation. 5 Intracellular CTLA-4 expression Is sustained In the presence of CD45RB/RO binding chimeric antibody It has been reported that substantial amounts of CTLA-4 can also be found intracellularly. Therefore, in parallel to surface CTLA-4 staining, intracellular CTLA-4 expression was analyzed. Moderate differences between T cell cultures were seen on day 4 after stimulation. 10 After prolonged culture, however, high levels of intracellular CTLA-4 were sustained only in CD45RB/RO binding chimeric antibody-treated but not in control T cells. CD45RB/RO binding chimeric antibody -treated T cells become double positive for CD4 and CD8 15 Following stimulation, T cells induce and upregulate the expression of several surface receptors, such as CD25, CD152 (CTLA-4), CD154 (CD40-Ligand) and others. In contrast, the level of expression of CD4 or CD8 is thought to stay relatively constant. We reproducibly observed a strong increase of both CD4 and CD8 antigens on CD45RB/RO binding chimeric antibody-treated but not on control Ab-treated T cells after activation. The emergence of a 20 CD4/CD8 double-positive T cell population seems to be due to the upregulation of CD4 on the CD8+ subset and conversely, CD8 on the CD4+ subset. This contrasts with a moderately low percentage of double positive T cells in control cultures. High IL-2 receptor alpha-chain, but very low beta-chain expression by CD45RB/RO 25 binding chimeric antibody-treated T cells Treg cells are known to be constitutively positive for CD25, the IL-2 receptor alpha-chain. The regulation of other subunits of the trimeric IL-2 receptor on Treg cells is not known. Recently we have compared the expression of the beta-chain of IL-2 receptor, e.g. CD122, on T cells activated and propagated In the presence or absence of CD45RB/RO binding 30 chimeric antibody. The results show that CD45RB/RO binding chimeric antibody-treated T cells have about ten-fold lower CD122 expression as compared to T cells in control cultures. This difference may indicate that Treg cells require factors other than IL-2 to proliferate. 35 -31 Example 8: Sequences of the invention (CDR sequences of the invention are underlined) 5 SEQ ID NO:1 Part of the amino acid sequence of chi ierc light chain DILLTQ SlAl'L VSPOEFV$FSCRSQNIGTSIQWYQQRTNGSPRLLIRSSSESISGIPSRFSG SGSGTDFTLSINSVESEDIADYYCQQSNTWPFTFGSGTKLEIK 10 SEQ ID NO:2 Part of the amino acid sequence of chime-c heavy chain EVQLQQSGPELVKPGASVKM'SCKASGYTFTNYIIHWVKQEPGQGLEWIGYFNPYNHGTKY NEKFKGRATLTADKSSNTAYMDLSSLTSEDSAIYYCARSGPYAWFDTWGQGTTVTVSS 15 SEQ ID NO:3 Amino acid sequence of chimeric light chain DILLTQSPAILSVSPGERVSFSCRASQNIGTSIQWYQQRTNGSPRLLIRSSSESISGIPSRFSG SGSGTDFTLSINSVESEDIADYYCQQSNTWPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKS GTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE 20 KHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO:4 Amino acid sequence of chimeric heavy chain EVQLQQSGPELVKPGASVKMSCKASGYTFTNYIIHWVKQEPGQGLEWIGYFNPYNHGTKY 25 NEKFKGRATLTADKSSNTAYMDLSSLTSEDSAIYYCARSGPYAWFDTWGQGTTVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTQTYCNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFL FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS 30 LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGK SEQ ID NO:5 Nucleotide sequence encoding a polypeptide of SEQ ID NO:1 - 32 GACATTCTGCTGACCCAGTCTCCAGCCATCCTGTCTGTGAGTCCAGGAGAGAGTCA GTTTCTCCTGCAGGGCCAGTCAGACATGGCACAGCATACAGTGGTATCACAGA ACAAATGGTTCTCCAAGGCTTCTCATAAGGTCTTCTCTGAGTCTATCTCTGGGATCCCT 5 TCCAGGTUrAGTGGCAGTGGATCAGGGACAGAT~TIACTCTTAGCATCACAGTGTGGA GTCTGMAGATATTGCAGA1-rAUTACTGTCACAGTMTACCTG GCCATTCACGTTCGG CTCG GGGACCAAGCTTGAAATCAAA SEQ ID NO:6 10 Nucleotide sequence encoding a polypeptide of SEQ ID NO:2 GAGGTGCAGCTGCAGCAGTCAGGACCTGMACTGGTWAGCCTGGGGCTTCAGTGMAG ATGTCCTGCMAGGCCTCTGGATACACATCACTMIAI.ATATTATCCACTG GGTGMAGCA GGAGCCTGGTCAGGGCCTTGMATGGAUTGGATAUUMTATCCUTACMTCATG GTACTA 15 AGTACAATGAGAAGTCAAAGGCAGGGCCACACTCTGCAGACATCCTCCMACACA GCCTACATGGACCTCAGCAGCCTGACCTCT GAGGACTCTGCGATCTACTACTGTGCMA GATCAGGACCCTATG CCTG GTTTGACACCTGGGGCCMAGGGACCACGGTCACCGTCTC CTCA 20 SEQ ID NO:7 Part of amino acid sequence of humanised light chain designated humV2 (humV2 Vim) DILLTQSPAT LSLSPGERAT FSCRASQNIGTSIQWYQQKT NGAPRLLIRS SSESISGIPS 25 RFSGSGSGTD FTLTISSLEP EDFAVYYCQQ.NTWpFTFGO GTKLEIK SEQ ID NO:8 Part of amino acid sequence of humanised light chain designated humnVl (humVl VLh) 30 DiLLTQSPAT LSLSPGERAT LSCRSN GTQWQQKP GQAPRLLIRS SSESISGI PS RFSGSGSGTD FTLTiSSLEP EDFAVYYCQQ SNTWPFTFGQ GTKLE 1K SEQ ID NO:9 -33 Part of amino acid sequence of humanised heavy chain designated VHE EVQLVESGAE VKKPGASVKV SCKASGYTFT NYIIHWVKQE PGQGLEWIGY FNPYNHGTKY NEKFKGRATL TANKSISTAY MELSSLRSED TAVYYCARSG 5 PYAWFDTWGQ GTTVTVSS SEQ ID NO:10 Part of amino acid sequence of humanised heavy chain designated VHQ 10 QVQLVESGAE VKKPGASVKV SCKASGYTFT NYIIHWVKQE PGQGLEWIGY FNPYNHGTKY NEKFKGRATL TANKSISTAY MELSSLRSED TAVYYCARSG PYAWFDTWGQ GTTVTVSS SEQ ID NO:11 15 Nucleotide sequence encoding amino acid sequence SEQ ID NO:9 GAGGTGCAGCTGGTGGAGTCAGGAGCCGAAGTGAAAAAGCCTGGGGCTTCAGTGAAG GTGTCCTGCAAGGCCTCTGGATACACATTCACTAATTATATTATCCACTGGGTGAAGCA GGAGCCTGGTCAGGGCCTTGAATGGATTGGATATTTTAATCCTTACAATCATG GTACTA 20 AGTACAATGAGAAGTTCAAAGGCAGGGCCACACTAACTGCAAACAAATCCATCAGCACA GCCTACATGGAGCTCAGCAGCCTGCGCTCTGAGGACACTGCGGTCTACTACTGTGCAA GATCAGGACCCTATGCCTGGTTTGACACCTGGGGCCAAGGGACCACGGTCACCGTCTC CTCA 25 SEQ ID NO:12 Nucleotide sequence encoding amino acid sequence SEQ ID NO:10 CAGGTGCAGCTGGTGGAGTCAGGAGCCGAAGTGAAAAAGCCTGGGGCTTCAGTGAAG GTGTCCTGCAAGGCCTCTGGATACACATTCACTAATTATATTATCCACTGGGTGAAGCA 30 GGAGCCTGGTCAGGGCCTTGAATGGATTGGATATTTTAATCCTTACAATCATGGTACTA AGTACAATGAGAAGTTCAAAGGCAGGGCCACACTAACTGCAAACAAATCCATCAGCACA GCCTACATGGAGCTCAGCAGCCTGCGCTCTGAGGACACTGCGGTCTACTACTGTGCAA GATCAGGACCCTATGCCTGGTTTGACACCTGGGGCCAAGGGACCACGGTCACCGTCTC

CTCA

-34 SEQ ID NO:13 Nucleotide sequence encoding amino acid sequence SEQ ID NO:7 5 GACAT1CTGCTGACCCAGTCTCCAGCCACCCTGTCTCTGAGTCCAGGAGAAAGAGCCA CTTTCTCCTGCAGGGCCAGTCAGAACATTGGCACAAGCATACAGTGGTATCAACAAAAA ACAAATGGTGCTCCAAGGCTTCTCATAAGGTCTTCTTCTGAGTCTATCT CTGGGATCCC TTCCAGGTTTAGTGGCAGTGGATCAGGGACAGA1TrTACTCTTACCATCAGCAGT CTGG AGCCTGAAGATTGCAGTGTATTACTGTCAACAAAGTAATACCTGGCCATTCACGTTC 10 GGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID NO:14 Nucleotide sequence encoding amino acid sequence SEQ ID NO:8 15 GACATTCTGCTGACCCAGTCTCCAGCCACCCTGTCTCTGAGTCCAGGAGAAAGAGCCA CTCTCTCCTGCAGGGCCAGTCAGAACATTGGCACAAGCATACAGTGGTATCAACAAAAA CCAGGTCAGGCTCCAAGGCTTCTCATAAGGTCTTCTTCTGAGTCTATCTCTGGGATCCC TTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTACCATCAGCAGT CTGG AGCCTGAAGATTTTGCAGTGTATTACTGTCAACAAAGTAATACCTGGCCATTCACGTTC 20 GGCCAGGGGACCAAGCTGGAGATCAAA SEQ ID NO:15 Nucleotide sequence of the expression vector HCMV-GI HuAb-VHQ (Complete DNA Sequence of a humanised heavy chain expression vector comprising 25 SEQ ID NO:12 (VHQ) from 3921-4274) 1 AGC7TTTTGC AAAAGCCTAG GCCTCCAAAA AAGCCTCCTC ACTACTTCTG 51 GAATAGCTCA GAGGCCGAGG CGGCCTCGGC CTCTGCATAA ATAAAAAAAA 101 TTAGTCAGCC ATGGGGCGGA GAATGGGCGG AACTGGGCGG AGTAGGGGC 30 151 GGGATGGGCG GAGTTAGGGG CGGGACTATG GTTGCTGACT AATTGAGATG 201 CATGCTTTGC ATACTTCTGC CTGCTGGGGA GCCTGGTTGC TGACTAATTG 251 AGATGCATGC TTTGCATACT TCTGCCTGCT GGGGAGCCTG GGGACTTTCC 301 ACACCCTAAC TGACACACAT TCCACAGCTG CCTCGCGCGT TTCGGTGATG 351 ACGGTGAAAA CCTCTGACAC ATGCAGCTCC CGGAGACGGT CACAGCTrGT -35 401 CTGTAAGCGG ATGCCGGGAG CAGACAAGCC CGTCAGGGCG CGTCAGCGGG 451 TGTTGGCGGG TGTCGGGGCG CAGCCATGAC CCAGTCACGT AGCGATAGCG 501 GAGTGTATAC TGGCTTAACT ATGCGGCATC AGAGCAGATT GTACTGAGAG 551 TGCACCATAT GCGGTGTGAA ATACCGCACA GATGCGTAAG GAGAAAATAC 5 601 CGCATCAGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT 651 CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT 701 TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC 751 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTrTTTCCA 801 TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA 10 851 GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA 901 AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT 951 GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT AGCTCACGCT 1001 GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG 1051 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG 15 1101 TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA 1151 CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC 1201 TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGGACAG TATTTGGTAT 1251 CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT GGTAGCTCTT 1301 GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTT TGTTTGCAAG 20 1351 CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT 1401 TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT 1451 TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA 1501 AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA 1551 CAGTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT 25 1601 TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT AACTACGATA 1651 CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC 1701 ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG 1751 CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT 1801 AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG 30 1851 CAACGTTGTT GCCATTGCTG CAGGCATCGT GGTGTCACGC TCGTCGTTTG 1901 GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG AGTTACATGA 1951 TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT 2001 TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT

ATGGCAGCAC

-36 2051 TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT 2101 GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG 2151 TTGCTCTTGC CCGGCGTCAA CACGGGATAA TACCGCGCCA CATAGCAGAA 2201 CTTTAAAAGT GCTCATCATT GGAAAACGTT CTTCGGGGCG AAAACTCTCA 5 2251 AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC 2301 CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA 2351 AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA 2401 TGTrGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA 2451 GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA 10 2501 AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC ACCTGACGTC 2551 TAAGAAACCA TTATTATCAT GACATTAACC TATAAAAATA GGCGTATCAC 2601 GAGGCCCTTT CGTCTTCAAG AATTCAGCTT GGCTGCAGTG AATAATAAAA 2651 TGTGTGTTTG TCCGAAATAC GCGTTTTGAG ATTTCTGTCG CCGACTAAAT 2701 TCATGTCGCG CGATAGTGGT GTTTATCGCC GATAGAGATG GCGATATTGG 15 2751 AAAAATCGAT ATTTGAAAAT ATGGCATATT GAAAATGTCG CCGATGTGAG 2801 TTTCTGTGTA ACTGATATCG CCAITTTTCC AAAAGTGATT TTTGGGCATA 2851 CGCGATATCT GGCGATAGCG CTTATATCGT TTACGGGGGA TGGCGATAGA 2901 CGACTTTGGT GACTTGGGCG ATTCTGTGTG TCGCAAATAT CGCAGTTTCG 2951 ATATAGGTGA CAGACGATAT GAGGCTATAT CGCCGATAGA GGCGACATCA 20 3001 AGCTGGCACA TGGCCAATGC ATATCGATCT ATACATTGAA TCAATATTGG 3051 CCATTAGCCA TArrATTCAT TGGTTATATA GCATAAATCA ATATTGGCTA 3101 TrGGCCATTG CATACGTTGT ATCCATATCA TAATATGTAC ATTTATATTG 3151 GCTCATGTCC AACATTACCG CCATGTTGAC ATTGATTATT GACTAGTTAT 3201 TAATAGTAAT CAATTACGGG GTCATTAGTT CATAGCCCAT ATATGGAGTT 25 3251 CCGCGTTACA TAACTTACGG TAAATGGCCC GCCTGGCTGA CCGCCCAACG 3301 ACCCCCGCCC ATTGACGTCA ATAATGACGT ATGTTCCCAT AGTAACGCCA 3351 ATAGGGACTT TCCATTGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC 3401 CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTACG CCCCCTATTG 3451 ACGTCAATGA CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC 30 3501 TTATGGGACT TTCCTACTTG GCAGTACATC TACGTATTAG TCATCGCTAT 3551 TACCATGGTG ATGCGGTTr GGCAGTACAT CAATGGGCGT GGATAGCGGT 3601 TrGACTCACG GGGATTTCCA AGTCTCCACC CCATTGACGT CAATGGGAGT 3651 TTGTTTTGGC ACCAAAATCA ACGGGACTTT CCAAAATGTC GTAACAACTC -37 3701 CGCCCCATTG ACGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA 3751 TAAGCAGAGC TCGTTTAGTG AACCGTCAGA TCGCCTGGAG ACGCCATCCA 3801 CGCTGTTTTG ACCTCCATAG AAGACACCGG GACCGATCCA GCCTCCGCAA 3851 GCTTGCCGCC ACCATGGACT GGACCTGGAG GGTGTTCTGC CTGCTGGCCG 5 3901 TGGCCCCCGG CGCCCACAGC CAGGTGCAGC TGGTGGAGTC AGGAGCCGAA 3951 GTGAAAAAGC CTGGGGCTTC AGTGAAGGTG TCCTGCAAGG CCTCTGGATA 4001 CACATTCACT AATTATATTA TCCACTGGGT GAAGCAGGAG CCTGGTCAGG 4051 GCCTTGAATG GATTGGATAT TTTAATCCTT ACAATCATGG TACTAAGTAC 4101 AATGAGAAGT TCAAAGGCAG GGCCACACTA ACTGCAAACA AATCCATCAG 10 4151 CACAGCCTAC ATGGAGCTCA GCAGCCTGCG CTCTGAGGAC ACTGCGGTCT 4201 ACTACTGTGC AAGATCAGGA CCCTATGCCT GGTTTGACAC CTGGGGCCAA 4251 GGGACCACGG TCACCGTCTC CTCAGGTGAG TTCTAGAAGG ATCCCAAGCT 4301 AGCTTTCTGG GGCAGGCCAG GCCTGACCTT GGCTTTGGGG CAGGGAGGGG 4351 GCTAAGGTGA GGCAGGTGGC GCCAGCCAGG TGCACACCCA ATGCCCATGA 15 4401 GCCCAGACAC TGGACGCTGA ACCTCGCGGA CAGTTAAGAA CCCAGGGGCC 4451 TCTGCGCCCT GGGCCCAGCT CTGTCCCACA CCGCGGTCAC ATGGCACCAC 4501 CTCTCTTGCA GCCTCCACCA AGGGCCCATC GGTCTTCCCC CTGGCACCCT 4551 CCTCCAAGAG CACCTCTGGG GGCACAGCGG CCCTGGGCTG CCTGGTCAAG 4601 GACTACTrCC CCGAACCGGT GACGGTGTCG TGGAACTCAG GCGCCCTGAC 20 4651 CAGCGGCGTG CACACCTTCC CGGCTGTCCT ACAGTCCTCA GGACTCTACT 4701 CCCTCAGCAG CGTGGTGACC GTGCCCTCCA GCAGCTTGGG CACCCAGACC 4751 TACATCTGCA ACGTGAATCA CAAGCCCAGC AACACCAAGG TGGACAAGAA 4801 AGTTGGTGAG AGGCCAGCAC AGGGAGGGAG GGTGTCTGCT GGAAGCCAGG 4851 CTCAGCGCTC CTGCCTGGAC GCATCCCGGC TATGCAGCCC CAGTCCAGGG 25 4901 CAGCAAGGCA GGCCCCGTCT GCCTCTTCAC CCGGAGGCCT CTGCCCGCCC 4951 CACTCATGCT CAGGGAGAGG GTCTTCTGGC TTTTTCCCCA GGCTCTGGGC 5001 AGGCACAGGC TAGGTGCCCC TAACCCAGGC CCTGCACACA AAGGGGCAGG 5051 TGCTGGGCTC AGACCTGCCA AGAGCCATAT CCGGGAGGAC CCTGCCCCTG 5101 ACCTAAGCCC ACCCCAAAGG CCAAACTCTC CACTCCCTCA GCTCGGACAC 30 5151 CTTCTCTCCT CCCAGATTCC AGTAACTCCC AATCTTCTCT CTGCAGAGCC 5201 CAAATCTTGT GACAAAACTC ACACATGCCC ACCGTGCCCA GGTAAGCCAG 5251 CCCAGGCCTC GCCCTCCAGC TCAAGGCGGG ACAGGTGCCC TAGAGTAGCC 5301 TGCATCCAGG GACAGGCCCC AGCCGGGTGC TGACACGTCC ACCTCCATCT -38 5351 CTTCCTCAGC ACCTGAACTC CTGGGGGGAC CGTCAGTCTT CCTCTTCCCC 5401 CCAAAACCCA AGGACACCCT CATGATCTCC CGGACCCCTG AGGTCACATG 5451 CGTGGTGGTG GACGTGAGCC ACGAAGACCC TGAGGTCAAG TTCAACTGGT 5501 ACGTGGACGG CGTGGAGGTG CATAATGCCA AGACAAAGCC GCGGGAGGAG 5 5551 CAGTACAACA GCACGTACCG TGTGGTCAGC GTCCTCACCG TCCTGCACCA 5601 GGACTGGCTG AATGGCAAGG AGTACAAGTG CAAGGTCTCC AACAAAGCCC 5651 TCCCAGCCCC CATCGAGAAA ACCATCTCCA AAGCCAAAGG TGGGACCCGT 5701 GGGGTGCGAG GGCCACATGG ACAGAGGCCG GCTCGGCCCA CCCTCTGCCC 5751 TGAGAGTGAC CGCTGTACCA ACCTCTGTCC CTACAGGGCA GCCCCGAGAA 10 5801 CCACAGGTGT ACACCCTGCC CCCATCCCGG GATGAGCTGA CCAAGAACCA 5851 GGTCAGCCTG ACCTGCCTGG TCAAAGGCTT CTATCCCAGC GACATCGCCG 5901 TGGAGTGGGA GAGCAATGGG CAGCCGGAGA ACAACTACAA GACCACGCCT 5951 CCCGTGCTGG ACTCCGACGG CTCCTTCTTC CTCTACAGCA AGCTCACCGT 6001 GGACAAGAGC AGGTGGCAGC AGGGGAACGT CTTCTCATGC TCCGTGATGC 15 6051 ATGAGGCTCT GCACAACCAC TACACGCAGA AGAGCCTCTC CCTGTCTCCG 6101 GGTAAATGAG TGCGACGGCC GGCAAGCCCC CGCTCCCCGG GCTCTCGCGG 6151 TCGCACGAGG ATGCTTGGCA CGTACCCCCT GTACATACTT CCCGGGCGCC 6201 CAGCATGGAA ATAAAGCACC CAGCGCTGCC CTGGGCCCCT GCGAGACTGT 6251 GATGGTTCTT TCCACGGGTC AGGCCGAGTC TGAGGCCTGA GTGGCATGAG 20 6301 ATCTGATATC ATCGATGAAT TCGAGCTCGG TACCCGGGGA TCGATCCAGA 6351 CATGATAAGA TACATTGATG AGTTTGGACA AACCACAACT AGAATGCAGT 6401 GAAAAAAATG CTTTATTTGT GAAATTTGTG ATGCTATTGC TTTATTTGTA 6451 ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT GCATTCATTT 6501 TATGTTTCAG GTTCAGGGGG AGGTGTGGGA GGTITrTAA AGCAAGTAAA 25 6551 ACCTCTACAA ATGTGGTATG GCTGATTATG ATCTCTAGTC AAGGCACTAT 6601 ACATCAAATA TTCCTTATTA ACCCCTTTAC AAATTAAAAA GCTAAAGGTA 6651 CACAATTTTT GAGCATAGTT ATTAATAGCA GACACTCTAT GCCTGTGTGG 6701 AGTAAGAAAA AACAGTATGT TATGATTATA ACTGTTATGC CTACTTATAA 6751 AGGTTACAGA ATATTTCC ATAATTTTCT TGTATAGCAG TGCAGCTTTT 30 6801 TCCTrTGTGG TGTAAATAGC AAAGCAAGCA AGAGTTCTAT TACTAAACAC 6851 AGCATGACTC AAAAAACTTA GCAATTCTGA AGGAAAGTCC TTGGGGTCTT 6901 CTACCTTTCT CTTCTTTTTT GGAGGAGTAG AATGTGAGA GTCAGCAGTA 6951 GCCTCATCAT CACTAGATGG CATTTCTTCT GAGCAAAACA GGTTrTCCTC -39 7001 ATTAAAGGCA TTCCACCACT GCTCCCATrC ATCAGTTCCA TAGGTTGGAA 7051 TCTAAAATAC ACAAACAATT AGAATCAGTA GTTTAACACA TTATACACrr 7101 AAAAATTA TATTTACCTT AGAGCTTTAA ATCTCTGTAG GTAGTTTGTC 7151 CAATTATGTC ACACCACAGA AGTAAGGrrC CTTCACAAAG ATCCGGGACC 5 7201 AAAGCGGCCA TCGTGCCTCC CCACTCCTGC AGTTCGGGGG CATGGATGCG 7251 CGGATAGCCG CTGCTGGTTT CCTGGATGCC GACGGATTTG CACTGCCGGT 7301 AGAACTCCGC GAGGTCGTCC AGCCTCAGGC AGCAGCTGAA CCAACTCGCG 7351 AGGGGATCGA GCCCGGGGTG GGCGAAGAAC TCCAGCATGA GATCCCCGCG 7401 CTGGAGGATC ATCCAGCCGG CGTCCCGGAA AACGATTCCG AAGCCCAACC 10 7451 TTTCATAGAA GGCGGCGGTG GAATCGAAAT CTCGTGATGG CAGGTTGGGC 7501 GTCGCTTGGT CGGTCATTTC GAACCCCAGA GTCCCGCTCA GAAGAACTCG 7551 TCAAGAAGGC GATAGAAGGC GATGCGCTGC GAATCGGGAG CGGCGATACC 7601 GTAAAGCACG AGGAAGCGGT CAGCCCArrC GCCGCCAAGC TCTTCAGCAA 7651 TATCACGGGT AGCCAACGCT ATGTCCTGAT AGCGGTCCGC CACACCCAGC 15 7701 CGGCCACAGT CGATGAATCC AGAAAAGCGG CCATTTTCCA CCATGATATT 7751 CGGCAAGCAG GCATCGCCAT GGGTCACGAC GAGATCCTCG CCGTCGGGCA 7801 TGCGCGCCTT GAGCCTGGCG AACAGTTCGG CTGGCGCGAG CCCCTGATGC 7851 TCTTCGTCCA GATCATCCTG ATCGACAAGA CCGGCTTCCA TCCGAGTACG 7901 TGCTCGCTCG ATGCGATGTT TCGCTTGGTG GTCGAATGGG CAGGTAGCCG 20 7951 GATCAAGCGT ATGCAGCCGC CGCATTGCAT CAGCCATGAT GGATACTTTC 8001 TCGGCAGGAG CAAGGTGAGA TGACAGGAGA TCCTGCCCCG GCACTTCGCC 8051 CAATAGCAGC CAGTCCCTTC CCGCTTCAGT GACAACGTCG AGCACAGCTG 8101 CGCAAGGAAC GCCCGTCGTG GCCAGCCACG ATAGCCGCGC TGCCTCGTCC 8151 TGCAGTTCAT TCAGGGCACC GGACAGGTCG GTCTTGACAA AAAGAACCGG 25 8201 GCGCCCCTGC GCTGACAGCC GGAACACGGC GGCATCAGAG CAGCCGATTG 8251 TCTGTTGTGC CCAGTCATAG CCGAATAGCC TCTCCACCCA AGCGGCCGGA 8301 GAACCTGCGT GCAATCCATC TTGTrCAATC ATGCGAAACG ATCCTCATCC 8351 TGTCTCTTGA TCAGATCTTG ATCCCCTGCG CCATCAGATC CTTGGCGGCA 8401 AGAAAGCCAT CCAGTTTACT TTGCAGGGCT TCCCAACCTT ACCAGAGGGC 30 8451 GCCCCAGCTG GCAATTCCGG TTCGCTTGCT GTCCATAAAA CCGCCCAGTC 8501 TAGCTATCGC CATGTAAGCC CACTGCAAGC TACCTGCTTT CTCTTTGCGC 8551 TTGCGTTrC CCTTGTCCAG ATAGCCCAGT AGCTGACATT CATCCGGGGT 8601 CAGCACCGTT TCTGCGGACT GGCTTTCTAC GTGTTCCGCT

TCCTTTAGCA

-40 8651 GCCCTTGCGC CCTGAGTGCT TGCGGCAGCG TGAAGCT SEQ ID NO:16 Nucleotide sequence of the expression vector HCMV-GI HuAb-VHE 5 (Complete DNA Sequence of a humanised heavy chain expression vector comprising SEQ ID NO: 11 (VHE) from 3921-4274) 1 AGCTTTTGC AAAAGCCTAG GCCTCCAAAA AAGCCTCCTC ACTACTTCTG 51 GAATAGCTCA GAGGCCGAGG CGGCCTCGGC CTCTGCATAA ATAAAAAAAA 10 101 TTAGTCAGCC ATGGGGCGGA GAATGGGCGG AACTGGGCGG AGTrAGGGGC 151 GGGATGGGCG GAGTTAGGGG CGGGACTATG GTTGCTGACT AATTGAGATG 201 CATGCTTTGC ATACTTCTGC CTGCTGGGGA GCCTGGTTGC TGACTAATTG 251 AGATGCATGC TTTGCATACT TCTGCCTGCT GGGGAGCCTG GGGACTTTCC 301 ACACCCTAAC TGACACACAT TCCACAGCrG CCTCGCGCGT TTCGGTGATG 15 351 ACGGTGAAAA CCTCTGACAC ATGCAGCTCC CGGAGACGGT CACAGCTTGT 401 CTGTAAGCGG ATGCCGGGAG CAGACAAGCC CGTCAGGGCG CGTCAGCGGG 451 TGTTGGCGGG TGTCGGGGCG CAGCCATGAC CCAGTCACGT AGCGATAGCG 501 GAGTGTATAC TGGCTTAACT ATGCGGCATC AGAGCAGATT GTACTGAGAG 551 TGCACCATAT GCGGTGTGAA ATACCGCACA GATGCGTAAG GAGAAAATAC 20 601 CGCATCAGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT 651 CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT 701 TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC 751 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTITTCCA 801 TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA 25 851 GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA 901 AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT 951 GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTITCTCAT AGCTCACGCT 1001 GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG 1051 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG 30 1101 TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA 1151 CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC 1201 TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGGACAG TATTTGGTAT 1251 CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT

GGTAGCTCTT

-41 1301 GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG 1351 CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT 1401 TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT 1451 TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA 5 1501 AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA 1551 CAGTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT 1601 TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT AACTACGATA 1651 CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC 1701 ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG 10 1751 CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT 1801 AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG 1851 CAACGTTGTT GCCATTGCTG CAGGCATCGT GGTGTCACGC TCGTCGTrTG 1901 GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG AGTTACATGA 1951 TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT 15 2001 TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC 2051 TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT 2101 GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG 2151 TTGCTCTTGC CCGGCGTCAA CACGGGATAA TACCGCGCCA CATAGCAGAA 2201 CTTTAAAAGT GCTCATCAT GGAAAACGTT CTTCGGGGCG AAAACTCTCA 20 2251 AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC 2301 CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA 2351 AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA 2401 TGTTGAATAC TCATACTCTT CCTTITCAA TATTATTGAA GCATTTATCA 2451 GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA 25 2501 AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC ACCTGACGTC 2551 TAAGAAACCA TTATTATCAT GACATTAACC TATAAAAATA GGCGTATCAC 2601 GAGGCCCTTT CGTCTTCAAG AATTCAGCTT GGCTGCAGTG AATAATAAAA 2651 TGTGTGTTTG TCCGAAATAC GCGTTTrGAG ATTTCTGTCG CCGACTAAAT 2701 TCATGTCGCG CGATAGTGGT GTTTATCGCC GATAGAGATG GCGATATTGG 30 2751 AAAAATCGAT ATTTGAAAAT ATGGCATATT GAAAATGTCG CCGATGTGAG 2801 TTTCTGTGTA ACTGATATCG CCATTTTTCC AAAAGTGATT TTTGGGCATA 2851 CGCGATATCT GGCGATAGCG CTTATATCGT TTACGGGGGA TGGCGATAGA 2901 CGACTTTGGT GACTTGGGCG ATTCTGTGTG TCGCAAATAT CGCAGTTTCG -42 2951 ATATAGGTGA CAGACGATAT GAGGCTATAT CGCCGATAGA GGCGACATCA 3001 AGCTGGCACA TGGCCAATGC ATATCGATCT ATACATTGAA TCAATATTGG 3051 CCATTAGCCA TATTATTCAT TGGTTATATA GCATAAATCA ATATTGGCTA 3101 TTGGCCATTG CATACGTTGT ATCCATATCA TAATATGTAC ATTTATATTG 5 3151 GCTCATGTCC AACATrACCG CCATGTTGAC ATTGATTATT GACTAGTTAT 3201 TAATAGTAAT CAATTACGGG GTCATTAGTT CATAGCCCAT ATATGGAGTT 3251 CCGCGTTACA TAACTTACGG TAAATGGCCC GCCTGGCTGA CCGCCCAACG 3301 ACCCCCGCCC ATTGACGTCA ATAATGACGT ATGTTCCCAT AGTAACGCCA 3351 ATAGGGACTT TCCATrGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC 10 3401 CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTACG CCCCCTATTG 3451 ACGTCAATGA CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC 3501 TTATGGGACT TTCCTACTTG GCAGTACATC TACGTATTAG TCATCGCTAT 3551 TACCATGGTG ATGCGGTTTT GGCAGTACAT CAATGGGCGT GGATAGCGGT 3601 TTGACTCACG GGGATTTCCA AGTCTCCACC CCATTGACGT CAATGGGAGT 15 3651 TTGTTTTGGC ACCAAAATCA ACGGGACT CCAAAATGTC GTAACAACTC 3701 CGCCCCATTG ACGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA 3751 TAAGCAGAGC TCGTTTAGTG AACCGTCAGA TCGCCTGGAG ACGCCATCCA 3801 CGCTGTTTTG ACCTCCATAG AAGACACCGG GACCGATCCA GCCTCCGCAA 3851 GCTTGCCGCC ACCATGGACT GGACCTGGAG GGTGTTCTGC CTGCTGGCCG 20 3901 TGGCCCCCGG CGCCCACAGC GAGGTGCAGC TGGTGGAGTC AGGAGCCGAA 3951 GTGAAAAAGC CTGGGGCTTC AGTGAAGGTG TCCTGCAAGG CCTCTGGATA 4001 CACATTCACT AATTATATTA TCCACTGGGT GAAGCAGGAG CCTGGTCAGG 4051 GCCTTGAATG GATTGGATAT TTTAATCCTT ACAATCATGG TACTAAGTAC 4101 AATGAGAAGT TCAAAGGCAG GGCCACACTA ACTGCAAACA AATCCATCAG 25 4151 CACAGCCTAC ATGGAGCTCA GCAGCCTGCG CTCTGAGGAC ACTGCGGTCT 4201 ACTACTGTGC AAGATCAGGA CCCTATGCCT GGTrTGACAC CTGGGGCCAA 4251 GGGACCACGG TCACCGTCTC CTCAGGTGAG TTCTAGAAGG ATCCCAAGCT 4301 AGCTTTCTGG GGCAGGCCAG GCCTGACCTT GGCTTTGGGG CAGGGAGGGG 4351 GCTAAGGTGA GGCAGGTGGC GCCAGCCAGG TGCACACCCA ATGCCCATGA 30 4401 GCCCAGACAC TGGACGCTGA ACCTCGCGGA CAGTTAAGAA CCCAGGGGCC 4451 TCTGCGCCCT GGGCCCAGCT CTGTCCCACA CCGCGGTCAC ATGGCACCAC 4501 CTCTCTTGCA GCCTCCACCA AGGGCCCATC GGTCTICCCC CTGGCACCCT 4551 CCTCCAAGAG CACCTCTGGG GGCACAGCGG CCCTGGGCTG CCTGGTCAAG -43 4601 GACTACTTCC CCGAACCGGT GACGGTGTCG TGGAACTCAG GCGCCCTGAC 4651 CAGCGGCGTG CACACCTTCC CGGCTGTCCT ACAGTCCTCA GGACTCTACT 4701 CCCTCAGCAG CGTGGTGACC GTGCCCTCCA GCAGCTTGGG CACCCAGACC 4751 TACATCTGCA ACGTGAATCA CAAGCCCAGC AACACCAAGG TGGACAAGAA 5 4801 AGTTGGTGAG AGGCCAGCAC AGGGAGGGAG GGTGTCTGCT GGAAGCCAGG 4851 CTCAGCGCTC CTGCCTGGAC GCATCCCGGC TATGCAGCCC CAGTCCAGGG 4901 CAGCAAGGCA GGCCCCGTCT GCCTCTTCAC CCGGAGGCCT CTGCCCGCCC 4951 CACTCATGCT CAGGGAGAGG GTCTTCTGGC TTTTTCCCCA GGCTCTGGGC 5001 AGGCACAGGC TAGGTGCCCC TAACCCAGGC CCTGCACACA AAGGGGCAGG 10 5051 TGCTGGGCTC AGACCTGCCA AGAGCCATAT CCGGGAGGAC CCTGCCCCTG 5101 ACCTAAGCCC ACCCCAAAGG CCAAACTCTC CACTCCCTCA GCTCGGACAC 5151 CTTCTCTCCT CCCAGATTCC AGTAACTCCC AATCTTCTCT CTGCAGAGCC 5201 CAAATCTTGT GACAAAACTC ACACATGCCC ACCGTGCCCA GGTAAGCCAG 5251 CCCAGGCCTC GCCCTCCAGC TCAAGGCGGG ACAGGTGCCC TAGAGTAGCC 15 5301 TGCATCCAGG GACAGGCCCC AGCCGGGTGC TGACACGTCC ACCTCCATCT 5351 CTTCCTCAGC ACCTGAACTC CTGGGGGGAC CGTCAGTCTT CCTCTTCCCC 5401 CCAAAACCCA AGGACACCCT CATGATCTCC CGGACCCCTG AGGTCACATG 5451 CGTGGTGGTG GACGTGAGCC ACGAAGACCC TGAGGTCAAG TTCAACTGGT 5501 ACGTGGACGG CGTGGAGGTG CATAATGCCA AGACAAAGCC GCGGGAGGAG 20 5551 CAGTACAACA GCACGTACCG TGTGGTCAGC GTCCTCACCG TCCTGCACCA 5601 GGACTGGCTG AATGGCAAGG AGTACAAGTG CAAGGTCTCC AACAAAGCCC 5651 TCCCAGCCCC CATCGAGAAA ACCATCTCCA AAGCCAAAGG TGGGACCCGT 5701 GGGGTGCGAG GGCCACATGG ACAGAGGCCG GCTCGGCCCA CCCTCTGCCC 5751 TGAGAGTGAC CGCTGTACCA ACCTCTGTCC CTACAGGGCA GCCCCGAGAA 25 5801 CCACAGGTGT ACACCCTGCC CCCATCCCGG GATGAGCTGA CCAAGAACCA 5851 GGTCAGCCTG ACCTGCCTGG TCAAAGGCTT CTATCCCAGC GACATCGCCG 5901 TGGAGTGGGA GAGCAATGGG CAGCCGGAGA ACAACTACAA GACCACGCCT 5951 CCCGTGCTGG ACTCCGACGG CTCCTTCTTC CTCTACAGCA AGCTCACCGT 6001 GGACAAGAGC AGGTGGCAGC AGGGGAACGT CTTCTCATGC TCCGTGATGC 30 6051 ATGAGGCTCT GCACAACCAC TACACGCAGA AGAGCCTCTC CCTGTCTCCG 6101 GGTAAATGAG TGCGACGGCC GGCAAGCCCC CGCTCCCCGG GCTCTCGCGG 6151 TCGCACGAGG ATGCTTGGCA CGTACCCCCT GTACATACTT CCCGGGCGCC 6201 CAGCATGGAA ATAAAGCACC CAGCGCTGCC CTGGGCCCCT GCGAGACTGT -44 6251 GATGGTTCTT TCCACGGGTC AGGCCGAGTC TGAGGCCTGA GTGGCATGAG 6301 ATCTGATATC ATCGATGAAT TCGAGCTCGG TACCCGGGGA TCGATCCAGA 6351 CATGATAAGA TACATTGATG AGTTTGGACA AACCACAACT AGAATGCAGT 6401 GAAAAAAATG CTTTATTTGT GAAATTTGTG ATGCTATTGC TTrATTTGTA 5 6451 ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT GCATTCATTT 6501 TATGTTTCAG GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA AGCAAGTAAA 6551 ACCTCTACAA ATGTGGTATG GCTGATTATG ATCTCTAGTC AAGGCACTAT 6601 ACATCAAATA TTCCTTATTA ACCCCTrTAC AAATTAAAAA GCTAAAGGTA 6651 CACAATTTT GAGCATAGTT ATTAATAGCA GACACTCTAT GCCTGTGTGG 10 6701 AGTAAGAAAA AACAGTATGT TATGATTATA ACTGTTATGC CTACTTATAA 6751 AGGTTACAGA ATATTTTTCC ATAATTTTCT TGTATAGCAG TGCAGCTTTT 6801 TCCTTTGTGG TGTAAATAGC AAAGCAAGCA AGAGTTCTAT TACTAAACAC 6851 AGCATGACTC AAAAAACTTA GCAATTCTGA AGGAAAGTCC TTGGGGTCTT 6901 CTACCTTTCT CTTCTTTITT GGAGGAGTAG AATGTTGAGA GTCAGCAGTA 15 6951 GCCTCATCAT CACTAGATGG CATTTCTTCT GAGCAAAACA GGTTTTCCTC 7001 ATTAAAGGCA TTCCACCACT GCTCCCATTC ATCAGTTCCA TAGGTTGGAA 7051 TCTAAAATAC ACAAACAATT AGAATCAGTA GTTTAACACA TTATACACTT 7101 AAAAATrTA TATTTACCTT AGAGCTTTAA ATCTCTGTAG GTAGTTTGTC 7151 CAATTATGTC ACACCACAGA AGTAAGGTTC CTTCACAAAG ATCCGGGACC 20 7201 AAAGCGGCCA TCGTGCCTCC CCACTCCTGC AGTTCGGGGG CATGGATGCG 7251 CGGATAGCCG CTGCTGGTTT CCTGGATGCC GACGGATTTG CACTGCCGGT 7301 AGAACTCCGC GAGGTCGTCC AGCCTCAGGC AGCAGCTGAA CCAACTCGCG 7351 AGGGGATCGA GCCCGGGGTG GGCGAAGAAC TCCAGCATGA GATCCCCGCG 7401 CTGGAGGATC ATCCAGCCGG CGTCCCGGAA AACGATTCCG AAGCCCAACC 25 7451 TTTCATAGAA GGCGGCGGTG GAATCGAAAT CTCGTGATGG CAGGTTGGGC 7501 GTCGCTTGGT CGGTCATTTC GAACCCCAGA GTCCCGCTCA GAAGAACTCG 7551 TCAAGAAGGC GATAGAAGGC GATGCGCTGC GAATCGGGAG CGGCGATACC 7601 GTAAAGCACG AGGAAGCGGT CAGCCCATIC GCCGCCAAGC TCTTCAGCAA 7651 TATCACGGGT AGCCAACGCT ATGTCCTGAT AGCGGTCCGC CACACCCAGC 30 7701 CGGCCACAGT CGATGAATCC AGAAAAGCGG CCATTTTCCA CCATGATATT 7751 CGGCAAGCAG GCATCGCCAT GGGTCACGAC GAGATCCTCG CCGTCGGGCA 7801 TGCGCGCCTT GAGCCTGGCG AACAGTTCGG CTGGCGCGAG CCCCTGATGC 7851 TCTTCGTCCA GATCATCCTG ATCGACAAGA CCGGCTTCCA TCCGAGTACG -45 7901 TGCTCGCTCG ATGCGATGTr TCGCTTGGTG GTCGAATGGG CAGGTAGCCG 7951 GATCAAGCGT ATGCAGCCGC CGCATTGCAT CAGCCATGAT GGATACTTTC 8001 TCGGCAGGAG CAAGGTGAGA TGACAGGAGA TCCTGCCCCG GCACTTCGCC 8051 CAATAGCAGC CAGTCCCTTC CCGCTTCAGT GACAACGTCG AGCACAGCTG 5 8101 CGCAAGGAAC GCCCGTCGTG GCCAGCCACG ATAGCCGCGC TGCCTCGTCC 8151 TGCAGTTCAT TCAGGGCACC GGACAGGTCG GTCTTGACAA AAAGAACCGG 8201 GCGCCCCTGC GCTGACAGCC GGAACACGGC GGCATCAGAG CAGCCGATTG 8251 TCTGTTGTGC CCAGTCATAG CCGAATAGCC TCTCCACCCA AGCGGCCGGA 8301 GAACCTGCGT GCAATCCATC TTGTTCAATC ATGCGAAACG ATCCTCATCC 10 8351 TGTCTCTTGA TCAGATCTTG ATCCCCTGCG CCATCAGATC CTTGGCGGCA 8401 AGAAAGCCAT CCAGTTTACT TTGCAGGGCT TCCCAACCTT ACCAGAGGGC 8451 GCCCCAGCTG GCAATTCCGG TTCGCTTGCT GTCCATAAAA CCGCCCAGTC 8501 TAGCTATCGC CATGTAAGCC CACTGCAAGC TACCTGCTTT CTCTTTGCGC 8551 TTGCGTTTTC CCTTGTCCAG ATAGCCCAGT AGCTGACATT CATCCGGGGT 15 8601 CAGCACCGTT TCTGCGGACT GGCTTTCTAC GTGTTCCGCT TCCTrTAGCA 8651 GCCCTTGCGC CCTGAGTGCT TGCGGCAGCG TGAAGCT SEQ ID NO:17 Nucleotide sequence of the expression vector HCMV-K HuAb-VLI hum VI 20 (Complete DNA Sequence of a humanised light chain expression vector comprising SEQ ID NO: 14 (humV1=VLh) from 3964-4284 1 CTAGCTTTTT GCAAAAGCCT AGGCCTCCAA AAAAGCCTCC TCACTACTTC 51 TGGAATAGCT CAGAGGCCGA GGCGGCCTCG GCCTCTGCAT AAATAAAAAA 25 101 AATTAGTCAG CCATGGGGCG GAGAATGGGC GGAACTGGGC GGAGTTAGGG 151 GCGGGATGGG CGGAGTTAGG GGCGGGACTA TGGTTGCTGA CTAATTGAGA 201 TGCATGCTTT GCATACTTCT GCCTGCTGGG GAGCCTGGTT GCTGACTAAT 251 TGAGATGCAT GCTTTGCATA CTTCTGCCTG CTGGGGAGCC TGGGGACTTr 301 CCACACCCTA ACTGACACAC ATTCCACAGC TGCCTCGCGC GTTTCGGTGA 30 351 TGACGGTGAA AACCTCTGAC ACATGCAGCT CCCGGAGACG GTCACAGCTT 401 GTCTGTAAGC GGATGCCGGG AGCAGACAAG CCCGTCAGGG CGCGTCAGCG 451 GGTGTTGGCG GGTGTCGGGG CGCAGCCATG ACCCAGTCAC GTAGCGATAG 501 CGGAGTGTAT ACTGGCTTAA CTATGCGGCA TCAGAGCAGA TTGTACTGAG -46 551 AGTGCACCAT ATGCGGTGTG AAATACCGCA CAGATGCGTA AGGAGAAAAT 601 ACCGCATCAG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG 651 GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG 701 GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG 5 751 GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC 801 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA 851 GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG 901 GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC 951 CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG 10 1001 CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG 1051 TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT 1101 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC 1151 CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT 1201 TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGGAC AGTATTTGGT 15 1251 ATCTGCGCTC TGCTGAAGCC AGTTACCTIC GGAAAAAGAG TTGGTAGCTC 1301 TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA 1351 AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC 1401 TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT 1451 TTTGGTCATG AGATTATCAA AAAGGATCT CACCTAGATC CTTTTAAATT 20 1501 AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT 1551 GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT 1601 ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA 1651 TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC 1701 CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG 25 1751 GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA 1801 TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG 1851 CGCAACGTTG TTGCCATTGC TGCAGGCATC GTGGTGTCAC GCTCGTCGTT 1901 TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT 1951 GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC 30 2001 GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC 2051 ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA 2101 CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG 2151 AGTTGCTCTT GCCCGGCGTC AACACGGGAT AATACCGCGC CACATAGCAG -47 2201 AACTTTAAAA GTGCTCATCA TTGGAAAACG TrCTTCGGGG CGAAAACTCT 2251 CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA 2301 CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC 2351 AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA 5 2401 AATGTTGAAT ACTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT 2451 CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA 2501 TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG 2551 TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC 2601 ACGAGGCCCT TTCGTCTTCA AGAATTCAGC TTGGCTGCAG TGAATAATAA 10 2651 AATGTGTGTT TGTCCGAAAT ACGCGTTTTG AGATTTCTGT CGCCGACTAA 2701 ATTCATGTCG CGCGATAGTG GTGTTTATCG CCGATAGAGA TGGCGATATT 2751 GGAAAAATCG ATATTTGAAA ATATGGCATA TTGAAAATGT CGCCGATGTG 2801 AGTTCTGTG TAACTGATAT CGCCATTTTT CCAAAAGTGA TTTTTGGGCA 2851 TACGCGATAT CTGGCGATAG CGCTTATATC GTTTACGGGG GATGGCGATA 15 2901 GACGACTTG GTGACTTGGG CGATTCTGTG TGTCGCAAAT ATCGCAGTTT 2951 CGATATAGGT GACAGACGAT ATGAGGCTAT ATCGCCGATA GAGGCGACAT 3001 CAAGCTGGCA CATGGCCAAT GCATATCGAT CTATACATTG AATCAATATT 3051 GGCCATTAGC CATATTATTC ATTGGTTATA TAGCATAAAT CAATATTGGC 3101 TATTGGCCAT TGCATACGTT GTATCCATAT CATAATATGT ACATTTATAT 20 3151 TGGCTCATGT CCAACATTAC CGCCATGTTG ACATTGATTA TTGACTAGTT 3201 ATTAATAGTA ATCAATTACG GGGTCATTAG TTCATAGCCC ATATATGGAG 3251 TTCCGCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT GACCGCCCAA 3301 CGACCCCCGC CCATTGACGT CAATAATGAC GTATGTTCCC ATAGTAACGC 3351 CAATAGGGAC TTTCCATTGA CGTCAATGGG TGGAGTATTT ACGGTAAACT 25 3401 GCCCACTTGG CAGTACATCA AGTGTATCAT ATGCCAAGTA CGCCCCCTAT 3451 TGACGTCAAT GACGGTAAAT GGCCCGCCTG GCATTATGCC CAGTACATGA 3501 CCTrATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT AGTCATCGCT 3551 ATTACCATGG TGATGCGGTr TTGGCAGTAC ATCAATGGGC GTGGATAGCG 3601 GTTTGACTCA CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA 30 3651 GTTTGTTTTG GCACCAAAAT CAACGGGACT TTCCAAAATG TCGTAACAAC 3701 TCCGCCCCAT TGACGCAAAT GGGCGGTAGG CGTGTACGGT GGGAGGTCTA 3751 TATAAGCAGA GCTCGTTTAG TGAACCGTCA GATCGCCTGG AGACGCCATC 3801 CACGCTGTTT TGACCTCCAT AGAAGACACC GGGACCGATC

CAGCCTCCGC

-48 3851 AAGCTTGATA TCGAATTCCT GCAGCCCGGG GGATCCGCCC GCTTGCCGCC 3901 ACCATGGAGA CCCCCGCCCA GCTGCTGTTC CTGCTGCTGC TGTGGCTGCC 3951 CGACACCACC GGCGACATTC TGCTGACCCA GTCTCCAGCC ACCCTGTCTC 4001 TGAGTCCAGG AGAAAGAGCC ACTCTCTCCT GCAGGGCCAG TCAGAACATT 5 4051 GGCACAAGCA TACAGTGGTA TCAACAAAAA CCAGGTCAGG CTCCAAGGCT 4101 TCTCATAAGG TCTTCTTCTG AGTCTATCTC TGGGATCCCT TCCAGGTTTA 4151 GTGGCAGTGG ATCAGGGACA GATTTTACTC TTACCATCAG CAGTCTGGAG 4201 CCTGAAGATT TTGCAGTGTA TTACTGTCAA CAAAGTAATA CCTGGCCATT 4251 CACGTTCGGC CAGGGGACCA AGCTGGAGAT CAAACGTGAG TATTCTAGAA 10 4301 AGATCCTAGA ATTCTAAACT CTGAGGGGGT CGGATGACGT GGCCATTCTT 4351 TGCCTAAAGC ATTGAGTTTA CTGCAAGGTC AGAAAAGCAT GCAAAGCCCT 4401 CAGAATGGCT GCAAAGAGCT CCAACAAAAC AATTTAGAAC TTTATTAAGG 4451 AATAGGGGGA AGCTAGGAAG AAACTCAAAA CATCAAGATT TTAAATACGC 4501 TTCTTGGTCT CCTTGCTATA ATTATCTGGG ATAAGCATGC TGTTrTCTGT 15 4551 CTGTCCCTAA CATGCCCTGT GATTATCCGC AAACAACACA CCCAAGGGCA 4601 GAACTTTGTT ACTTAAACAC CATCCTGTTr GCTrCTTTCC TCAGGAACTG 4651 TGGCTGCACC ATCTGTCTTC ATCTTCCCGC CATCTGATGA GCAGTTGAAA 4701 TCTGGAACTG CCTCTGTTGT GTGCCTGCTG AATAACTTCT ATCCCAGAGA 4751 GGCCAAAGTA CAGTGGAAGG TGGATAACGC CCTCCAATCG GGTAACTCCC 20 4801 AGGAGAGTGT CACAGAGCAG GACAGCAAGG ACAGCACCTA CAGCCTCAGC 4851 AGCACCCTGA CGCTGAGCAA AGCAGACTAC GAGAAACACA AAGTCTACGC 4901 CTGCGAAGTC ACCCATCAGG GCCTGAGCTC GCCCGTCACA AAGAGCTTCA 4951 ACAGGGGAGA GTGTTAGAGG GAGAAGTGCC CCCACCTGCT CCTCAGTTCC 5001 AGCCTGACCC CCTCCCATCC TTTGGCCTCT GACCCTTTTT CCACAGGGGA 25 5051 CCTACCCCTA TTGCGGTCCT CCAGCTCATC TTTCACCTCA CCCCCCTCCT 5101 CCTCCTTGGC TTTAATTATG CTAATGTTGG AGGAGAATGA ATAAATAAAG 5151 TGAATCTTTG CACCTGTGGT TTCTCTCTTT CCTCATTTAA TAATTATTAT 5201 CTGTTGTTTA CCAACTACTC AATTTCTCTT ATAAGGGACT AAATATGTAG 5251 TCATCCTAAG GCGCATAACC ATTTATAAAA ATCATCCTTC ATTCTATTTT 30 5301 ACCCTATCAT CCTCTGCAAG ACAGTCCTCC CTCAAACCCA CAAGCCTTCT 5351 GTCCTCACAG TCCCCTGGGC CATGGTAGGA GAGACTTGCT TCCTTGTrTT 5401 CCCCTCCTCA GCAAGCCCTC ATAGTCCTTT TTAAGGGTGA CAGGTCTTAC 5451 AGTCATATAT CCTTTGATTC AATTCCCTGA GAATCAACCA AAGCAAATTT -49 5501 TTCAAAAGAA GAAACCTGCT ATAAAGAGAA TCATTCATrG CAACATGATA 5551 TAAAATAACA ACACAATAAA AGCAATAAA TAAACAAACA ATAGGGAAAT 5601 GTTTAAGTTC ATCATGGTAC TTAGACTTAA TGGAATGTCA TGCCTTATTT 5651 ACATTTTTAA ACAGGTACTG AGGGACTCCT GTCTGCCAAG GGCCGTATTG 5 5701 AGTACTTTCC ACAACCTAAT TTAATCCACA CTATACTGTG AGATTAAAAA 5751 CATTCATTAA AATGTTGCAA AGGTTCTATA AAGCTGAGAG ACAAATATAT 5801 TCTATAACTC AGCAATCCCA CTTCTAGATG ACTGAGTGTC CCCACCCACC 5851 AAAAAACTAT GCAAGAATGT TCAAAGCAGC TTTATTTACA AAAGCCAAAA 5901 ATTGGAAATA GCCCGATTGT CCAACAATAG AATGAGTTAT TAAACTGTGG 10 5951 TATGTTTATA CATTAGAATA CCCAATGAGG AGAATTAACA AGCTACAACT 6001 ATACCTACTC ACACAGATGA ATCTCATAAA AATAATGTTA CATAAGAGAA 6051 ACTCAATGCA AAAGATATGT TCTGTATGTT TrCATCCATA TAAAGTTCAA 6101 AACCAGGTAA AAATAAAGTT AGAAATTTGG ATGGAAATTA CTCTTAGCTG 6151 GGGGTGGGCG AGTTAGTGCC TGGGAGAAGA CAAGAAGGGG CTTCTGGGGT 15 6201 CTTGGTAATG TTCTGTTCCT CGTGTGGGGT TGTGCAGTTA TGATCTGTGC 6251 ACTGTTCTGT ATACACATTA TGCTTCAAAA TAACTTCACA TAAAGAACAT 6301 CTTATACCCA GTTAATAGAT AGAAGAGGAA TAAGTAATAG GTCAAGACCA 6351 CGCAGCTGGT AAGTGGGGGG GCCTGGGATC AAATAGCTAC CTGCCTAATC 6401 CTGCCCTCTT GAGCCCTGAA TGAGTCTGCC TTCCAGGGCT CAAGGTGCTC 20 6451 AACAAAACAA CAGGCCTGCT ATTTTCCTGG CATCTGTGCC CTGTTTGGCT 6501 AGCTAGGAGC ACACATACAT AGAAATTAAA TGAAACAGAC CTTCAGCAAG 6551 GGGACAGAGG ACAGAATTAA CCTTGCCCAG ACACTGGAAA CCCATGTATG 6601 AACACTCACA TGTTTGGGAA GGGGGAAGGG CACATGTAAA TGAGGACTCT 6651 TCCTCATTCT ATGGGGCACT CTGGCCCTGC CCCTCTCAGC TACTCATCCA 25 6701 TCCAACACAC CTTTCTAAGT ACCTCTCTCT GCCTACACTC TGAAGGGGTT 6751 CAGGAGTAAC TAACACAGCA TCCCTTCCCT CAAATGACTG ACAATCCCTT 6801 TGTCCTGCTT TGTTTTrCTT TCCAGTCAGT ACTGGGAAAG TGGGGAAGGA 6851 CAGTCATGGA GAAACTACAT AAGGAAGCAC CTTGCCCTTC TGCCTCTTGA 6901 GAATGTTGAT GAGTATCAAA TCTTTCAAAC TTTGGAGGTT TGAGTAGGGG 30 6951 TGAGACTCAG TAATGTCCCT TCCAATGACA TGAACTTGCT CACTCATCCC 7001 TGGGGGCCAA ATTGAACAAT CAAAGGCAGG CATAATCCAG CTATGAATTC 7051 TAGGATCGAT CCAGACATGA TAAGATACAT TGATGAGTIT GGACAAACCA 7101 CAACTAGAAT GCAGTGAAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT -50 7151 ATTGCTTTAT TTGTAACCAT TATAAGCTGC AATAAACAAG TTAACAACAA 7201 CAATTGCATT CATTTrATGT TTCAGGTTCA GGGGGAGGTG TGGGAGGTTT 7251 TTTAAAGCAA GTAAAACCTC TACAAATGTG GTATGGCTGA TTATGATCTC 7301 TAGTCAAGGC ACTATACATC AAATATTCCT TATTAACCCC TTTACAAATT 5 7351 AAAAAGCTAA AGGTACACAA T=TTGAGCA TAGTTATTAA TAGCAGACAC 7401 TCTATGCCTG TGTGGAGTAA GAAAAAACAG TATGTTATGA TTATAACTGT 7451 TATGCCTACT TATAAAGGTT ACAGAATATT TTTCCATAAT TTTCTTGTAT 7501 AGCAGTGCAG CTIrTTCCTT TGTGGTGTAA ATAGCAAAGC AAGCAAGAGT 7551 TCTATTACTA AACACAGCAT GACTCAAAAA ACTTAGCAAT TCTGAAGGAA 10 7601 AGTCCTTGGG GTCTTCTACC TTTCTCTTCT TTTTTGGAGG AGTAGAATGT 7651 TGAGAGTCAG CAGTAGCCTC ATCATCACTA GATGGCATTT CTTCTGAGCA 7701 AAACAGGTTT TCCTCATTAA AGGCATTCCA CCACTGCTCC CATrCATCAG 7751 TTCCATAGGT TGGAATCTAA AATACACAAA CAATTAGAAT CAGTAGTTTA 7801 ACACATTATA CACTTAAAAA ITTATATT ACCTTAGAGC TTTAAATCTC 15 7851 TGTAGGTAGT TTGTCCAATT ATGTCACACC ACAGAAGTAA GGTTCCTTCA 7901 CAAAGATCCG GGACCAAAGC GGCCATCGTG CCTCCCCACT CCTGCAGTTC 7951 GGGGGCATGG ATGCGCGGAT AGCCGCTGCT GGTTTCCTGG ATGCCGACGG 8001 ATTTGCACTG CCGGTAGAAC TCCGCGAGGT CGTCCAGCCT CAGGCAGCAG 8051 CTGAACCAAC TCGCGAGGGG ATCGAGCCCG GGGTGGGCGA AGAACTCCAG 20 8101 CATGAGATCC CCGCGCTGGA GGATCATCCA GCCGGCGTCC CGGAAAACGA 8151 TTCCGAAGCC CAACCTTTCA TAGAAGGCGG CGGTGGAATC GAAATCTCGT 8201 GATGGCAGGT TGGGCGTCGC TTGGTCGGTC ATTTCGAACC CCAGAGTCCC 8251 GCTCAGAAGA ACTCGTCAAG AAGGCGATAG AAGGCGATGC GCTGCGAATC 8301 GGGAGCGGCG ATACCGTAAA GCACGAGGAA GCGGTCAGCC CATTCGCCGC 25 8351 CAAGCTCTTC AGCAATATCA CGGGTAGCCA ACGCTATGTC CTGATAGCGG 8401 TCCGCCACAC CCAGCCGGCC ACAGTCGATG AATCCAGAAA AGCGGCCATT 8451 TTCCACCATG ATATTCGGCA AGCAGGCATC GCCATGGGTC ACGACGAGAT 8501 CCTCGCCGTC GGGCATGCGC GCCTTGACCC TGGCGAACAG TTCGGCTGGC 8551 GCGAGCCCCT GATGCTCTTC GTCCAGATCA TCCTGATCGA CAAGACCGGC 30 8601 TTCCATCCGA GTACGTGCTC GCTCGATGCG ATGTTTCGCT TGGTGGTCGA 8651 ATGGGCAGGT AGCCGGATCA AGCGTATGCA GCCGCCGCAT TGCATCAGCC 8701 ATGATGGATA CTTrCTCGGC AGGAGCAAGG TGAGATGACA GGAGATCCTG 8751 CCCCGGCACT TCGCCCAATA GCAGCCAGTC CCTTCCCGCT TCAGTGACAA -51 8801 CGTCGAGCAC AGCTGCGCAA GGAACGCCCG TCGTGGCCAG CCACGATAGC 8851 CGCGCTGCCT CGTCCTGCAG TTCATTCAGG GCACCGGACA GGTCGGTCTT 8901 GACAAAAAGA ACCGGGCGCC CCTGCGCTGA CAGCCGGAAC ACGGCGGCAT 8951 CAGAGCAGCC GATTGTCTGT TGTGCCCAGT CATAGCCGAA TAGCCTCTCC 5 9001 ACCCAAGCGG CCGGAGAACC TGCGTGCAAT CCATCTTGTT CAATCATGCG 9051 AAACGATCCT CATCCTGTCT CTTGATCAGA TCTTGATCCC CTGCGCCATC 9101 AGATCCTrGG CGGCAAGAAA GCCATCCAGT TTACTTTGCA GGGCTTCCCA 9151 ACCTTACCAG AGGGCGCCCC AGCTGGCAAT TCCGGTTCGC TTGCTGTCCA 9201 TAAAACCGCC CAGTCTAGCT ATCGCCATGT AAGCCCACTG CAAGCTACCT 10 9251 GCTTTCTCTT TGCGCTTGCG TTTTCCCTTG TCCAGATAGC CCAGTAGCTG 9301 ACATTCATCC GGGGTCAGCA CCGTtTCTGC GGACTGGCTT TCTACGTGTT 9351 CCGCTTCCTT TAGCAGCCCT TGCGCCCTGA GTGCTTGCGG CAGCGTGAAG SEQ ID NO:18 15 Nucleotide sequence of the expression vector HCMV-K HuAb-VLI hum V2 (Complete DNA Sequence of a humanised light chain expression vector comprising SEQ ID NO: 13 (humV2=VLm) from 3926-4246) 1 CTAGCTTTT GCAAAAGCCT AGGCCTCCAA AAAAGCCTCC TCACTACTTC 20 51 TGGAATAGCT CAGAGGCCGA GGCGGCCTCG GCCTCTGCAT AAATAAAAAA 101 AATTAGTCAG CCATGGGGCG GAGAATGGGC GGAACTGGGC GGAGTTAGGG 151 GCGGGATGGG CGGAGTTAGG GGCGGGACTA TGGTrGCTGA CTAATTGAGA 201 TGCATGCTTT GCATACTTCT GCCTGCTGGG GAGCCTGGTT GCTGACTAAT 251 TGAGATGCAT GCTTTGCATA CTTCTGCCTG CTGGGGAGCC TGGGGACTTT 25 301 CCACACCCTA ACTGACACAC ATTCCACAGC TGCCTCGCGC GTTTCGGTGA 351 TGACGGTGAA AACCTCTGAC ACATGCAGCT CCCGGAGACG GTCACAGCTT 401 GTCTGTAAGC GGATGCCGGG AGCAGACAAG CCCGTCAGGG CGCGTCAGCG 451 GGTGTTGGCG GGTGTCGGGG CGCAGCCATG ACCCAGTCAC GTAGCGATAG 501 CGGAGTGTAT ACTGGCTTAA CTATGCGGCA TCAGAGCAGA TTGTACTGAG 30 551 AGTGCACCAT ATGCGGTGTG AAATACCGCA CAGATGCGTA AGGAGAAAAT 601 ACCGCATCAG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG 651 GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG 701 GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG -52 751 GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC 801 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA 851 GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG 901 GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC 5 951 CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG 1001 CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG 1051 TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT 1101 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC 1151 CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT 10 1201 TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGGAC AGTATTTGGT 1251 ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC 1301 TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA 1351 AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC 1401 TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT 15 1451 TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT 1501 AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT 1551 GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT 1601 ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA 1651 TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC 20 1701 CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG 1751 GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA 1801 TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG 1851 CGCAACGTTG TTGCCATTGC TGCAGGCATC GTGGTGTCAC GCTCGTCGTT 1901 TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT 25 1951 GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC 2001 GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC 2051 ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA 2101 CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG 2151 AGTTGCTCTT GCCCGGCGTC AACACGGGAT AATACCGCGC CACATAGCAG 30 2201 AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT 2251 CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA 2301 CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC 2351 AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA -53 2401 AATGTTGAAT ACTCATACTC TTCCTTTTC AATATTATTG AAGCATTAT 2451 CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA 2501 TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG 2551 TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC 5 2601 ACGAGGCCCT TTCGTCTTCA AGAATTCAGC TrGGCTGCAG TGAATAATAA 2651 AATGTGTGTT TGTCCGAAAT ACGCGTTTTG AGATTTCTGT CGCCGACTAA 2701 ATTCATGTCG CGCGATAGTG GTGTTTATCG CCGATAGAGA TGGCGATATT 2751 GGAAAAATCG ATATTTGAAA ATATGGCATA TTGAAAATGT CGCCGATGTG 2801 AGTTTCTGTG TAACTGATAT CGCCATTTTT CCAAAAGTGA TTTTTGGGCA 10 2851 TACGCGATAT CTGGCGATAG CGCTTATATC GTTTACGGGG GATGGCGATA 2901 GACGACITTG GTGACTTGGG CGATTCTGTG TGTCGCAAAT ATCGCAGTTT 2951 CGATATAGGT GACAGACGAT ATGAGGCTAT ATCGCCGATA GAGGCGACAT 3001 CAAGCTGGCA CATGGCCAAT GCATATCGAT CTATACATrG AATCAATATT 3051 GGCCATTAGC CATATTATTC ATTGGTTATA TAGCATAAAT CAATATTGGC 15 3101 TATTGGCCAT TGCATACGTT GTATCCATAT CATAATATGT ACATTTATAT 3151 TGGCTCATGT CCAACATTAC CGCCATGTTG ACATTGATTA TTGACTAGTT 3201 ATTAATAGTA ATCAATTACG GGGTCATTAG TTCATAGCCC ATATATGGAG 3251 TTCCGCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT GACCGCCCAA 3301 CGACCCCCGC CCATTGACGT CAATAATGAC GTATGTTCCC ATAGTAACGC 20 3351 CAATAGGGAC TTTCCATTGA CGTCAATGGG TGGAGTATTT ACGGTAAACT 3401 GCCCACTTGG CAGTACATCA AGTGTATCAT ATGCCAAGTA CGCCCCCTAT 3451 TGACGTCAAT GACGGTAAAT GGCCCGCCTG GCATTATGCC CAGTACATGA 3501 CCTTATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT AGTCATCGCT 3551 ATTACCATGG TGATGCGGTT TTGGCAGTAC ATCAATGGGC GTGGATAGCG 25 3601 GTTTGACTCA CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA 3651 GTTTGTTTTG GCACCAAAAT CAACGGGACT TTCCAAAATG TCGTAACAAC 3701 TCCGCCCCAT TGACGCAAAT GGGCGGTAGG CGTGTACGGT GGGAGGTCTA 3751 TATAAGCAGA GCTCGTTTAG TGAACCGTCA GATCGCCTGG AGACGCCATC 3801 CACGCTGTTT TGACCTCCAT AGAAGACACC GGGACCGATC CAGCCTCCGC 30 3851 AAGCTTGCCG CCACCATGGA GACCCCCGCC CAGCTGCTGT TCCTGCTGCT 3901 GCTGTGGCTG CCCGACACCA CCGGCGACAT TCTGCTGACC CAGTCTCCAG 3951 CCACCCTGTC TCTGAGTCCA GGAGAAAGAG CCACTTTCTC CTGCAGGGCC 4001 AGTCAGAACA TTGGCACAAG CATACAGTGG TATCAACAAA AAACAAATGG -54 4051 TGCTCCAAGG CTTCTCATAA GGTCTTCTTC TGAGTCTATC TCTGGGATCC 4101 CTTCCAGGTT TAGTGGCAGT GGATCAGGGA CAGATTTTAC TCTTACCATC 4151 AGCAGTCTGG AGCCTGAAGA TrTGCAGTG TATTACTGTC AACAAAGTAA 4201 TACCTGGCCA TTCACGTTCG GCCAGGGGAC CAAGCTGGAG ATCAAACGTG 5 4251 AGTATTCTAG AAAGATCCTA GAATTCTAAA CTCTGAGGGG GTCGGATGAC 4301 GTGGCCATTC TTTGCCTAAA GCATTGAGTT TACTGCAAGG TCAGAAAAGC 4351 ATGCAAAGCC CTCAGAATGG CTGCAAAGAG CTCCAACAAA ACAATTTAGA 4401 ACTTTATTAA GGAATAGGGG GAAGCTAGGA AGAAACTCAA AACATCAAGA 4451 TTTTAAATAC GCTTCTTGGT CTCCTTGCTA TAATTATCTG GGATAAGCAT 10 4501 GCTGTTTrCT GTCTGTCCCT AACATGCCCT GTGATTATCC GCAAACAACA 4551 CACCCAAGGG CAGAACTTTG TTACTTAAAC ACCATCCTGT TTGCTTCTTT 4601 CCTCAGGAAC TGTGGCTGCA CCATCTGTCT TCATCTTCCC GCCATCTGAT 4651 GAGCAGTTGA AATCTGGAAC TGCCTCTGTT GTGTGCCTGC TGAATAACTT 4701 CTATCCCAGA GAGGCCAAAG TACAGTGGAA GGTGGATAAC GCCCTCCAAT 15 4751 CGGGTAACTC CCAGGAGAGT GTCACAGAGC AGGACAGCAA GGACAGCACC 4801 TACAGCCTCA GCAGCACCCT GACGCTGAGC AAAGCAGACT ACGAGAAACA 4851 CAAAGTCTAC GCCTGCGAAG TCACCCATCA GGGCCTGAGC TCGCCCGTCA 4901 CAAAGAGCTT CAACAGGGGA GAGTGTTAGA GGGAGAAGTG CCCCCACCTG 4951 CTCCTCAGTT CCAGCCTGAC CCCCTCCCAT CCTTTGGCCT CTGACCCTTT 20 5001 TTCCACAGGG GACCTACCCC TATTGCGGTC CTCCAGCTCA TCTTTCACCT 5051 CACCCCCCTC CTCCTCCTTG GCTTTAATTA TGCTAATGTT GGAGGAGAAT 5101 GAATAAATAA AGTGAATCTT TGCACCTGTG GTTTCTCTCT TTCCTCATTT 5151 AATAATTATT ATCTGTTGTT TACCAACTAC TCAATTTCTC TTATAAGGGA 5201 CTAAATATGT AGTCATCCTA AGGCGCATAA CCATTTATAA AAATCATCCT 25 5251 TCATTCTATT TTACCCTATC ATCCTCTGCA AGACAGTCCT CCCTCAAACC 5301 CACAAGCCTT CTGTCCTCAC AGTCCCCTGG GCCATGGTAG GAGAGACTTG 5351 CTTCCTTGTT TTCCCCTCCT CAGCAAGCCC TCATAGTCCT TTTTAAGGGT 5401 GACAGGTCTT ACAGTCATAT ATCCTTTGAT TCAATTCCCT GAGAATCAAC 5451 CAAAGCAAAT TTTTCAAAAG AAGAAACCTG CTATAAAGAG AATCATTCAT 30 5501 TGCAACATGA TATAAAATAA CAACACAATA AAAGCAATTA AATAAACAAA 5551 CAATAGGGAA ATGTTTAAGT TCATCATGGT ACTTAGACTT AATGGAATGT 5601 CATGCCTTAT TTACATIrTT AAACAGGTAC TGAGGGACTC CTGTCTGCCA 5651 AGGGCCGTAT TGAGTACTTT CCACAACCTA ATTTAATCCA

CACTATACTG

-55 5701 TGAGATrAAA AACATTCATT AAAATGTTGC AAAGGTTCTA TAAAGCTGAG 5751 AGACAAATAT ATTCTATAAC TCAGCAATCC CACTTCTAGA TGACTGAGTG 5801 TCCCCACCCA CCAAAAAACT ATGCAAGAAT GTTCAAAGCA GCITTATTTA 5851 CAAAAGCCAA AAATTGGAAA TAGCCCGATT GTCCAACAAT AGAATGAGTT 5 5901 ATTAAACTGT GGTATGTTTA TACATTAGAA TACCCAATGA GGAGAATTAA 5951 CAAGCTACAA CTATACCTAC TCACACAGAT GAATCTCATA AAAATAATGT 6001 TACATAAGAG AAACTCAATG CAAAAGATAT GTTCTGTATG TTTTCATCCA 6051 TATAAAGTTC AAAACCAGGT AAAAATAAAG TTAGAAATTT GGATGGAAAT 6101 TACTCTrAGC TGGGGGTGGG CGAGTTAGTG CCTGGGAGAA GACAAGAAGG 10 6151 GGCTTCTGGG GTCTTGGTAA TGTTCTGTTC CTCGTGTGGG GTTGTGCAGT 6201 TATGATCTGT GCACTGTTCT GTATACACAT TATGCTTCAA AATAACTTCA 6251 CATAAAGAAC ATCTTATACC CAGTTAATAG ATAGAAGAGG AATAAGTAAT 6301 AGGTCAAGAC CACGCAGCTG GTAAGTGGGG GGGCCTGCGA TCAAATAGCT 6351 ACCTGCCTAA TCCTGCCCTC TTGAGCCCTG AATGAGTCTG CCTTCCAGGG 15 6401 CTCAAGGTGC TCAACAAAAC AACAGGCCTG CTATTTTCCT GGCATCTGTG 6451 CCCTGTTTGG CTAGCTAGGA GCACACATAC ATAGAAATTA AATGAAACAG 6501 ACCTTCAGCA AGGGGACAGA GGACAGAATT AACCTTGCCC AGACACTGGA 6551 AACCCATGTA TGAACACTCA CATGTTTGGG AAGGGGGAAG GGCACATGTA 6601 AATGAGGACT CTTCCTCATT CTATGGGGCA CTCTGGCCCT GCCCCTCTCA 20 6651 GCTACTCATC CATCCAACAC ACCTTTCTAA GTACCTCTCT CTGCCTACAC 6701 TCTGAAGGGG TTCAGGAGTA ACTAACACAG CATCCCTTCC CTCAAATGAC 6751 TGACAATCCC TTTGTCCTGC TTTGTTTTTC TTTCCAGTCA GTACTGGGAA 6801 AGTGGGGAAG GACAGTCATG GAGAAACTAC ATAAGGAAGC ACCTTGCCCT 6851 TCTGCCTCTT GAGAATGTTG ATGAGTATCA AATCTTTCAA ACTTTGGAGG 25 6901 TTTGAGTAGG GGTGAGACTC AGTAATGTCC CTTCCAATGA CATGAACTTG 6951 CTCACTCATC CCTGGGGGCC AAATTGAACA ATCAAAGGCA GGCATAATCC 7001 AGCTATGAAT TCTAGGATCG ATCCAGACAT GATAAGATAC ATTGATGAGT 7051 TTGGACAAAC CACAACTAGA ATGCAGTGAA AAAAATGCTT TATTTGTGAA 7101 ATTTGTGATG CTATTGCTTT ATTTGTAACC ATTATAAGCT GCAATAAACA 30 7151 AGTTAACAAC AACAATTGCA TrCATTTTAT GTTTCAGGTT CAGGGGGAGG 7201 TGTGGGAGGT TTTTTAAAGC AAGTAAAACC TCTACAAATG TGGTATGGCT 7251 GATTATGATC TCTAGTCAAG GCACTATACA TCAAATATrC CTTATTAACC 7301 CCTTTACAAA TTAAAAAGCT AAAGGTACAC AATTrTGAG CATAGTTATr -56 7351 AATAGCAGAC ACTCTATGCC TGTGTGGAGT AAGAAAAAAC AGTATGTTAT 7401 GATTATAACT GTTATGCCTA CTTATAAAGG TTACAGAATA TTTTTCCATA 7451 ATTTTCTTGT ATAGCAGTGC AGCTTTTTCC TTGTGGTGT AAATAGCAAA 7501 GCAAGCAAGA GTTCTATrAC TAAACACAGC ATGACTCAAA AAACTTAGCA 5 7551 ATTCTGAAGG AAAGTCCTTG GGGTCTTCTA CCTTTCTCTT CTTTTTTGGA 7601 GGAGTAGAAT GTTGAGAGTC AGCAGTAGCC TCATCATCAC TAGATGGCAT 7651 TTCTTCTGAG CAAAACAGGT TTTCCTCATT AAAGGCATTC CACCACTGCT 7701 CCCATTCATC AGTTCCATAG GTTGGAATCT AAAATACACA AACAATTAGA 7751 ATCAGTAGTT TAACACATTA TACACTTAAA AATTTTATAT TTACCTTAGA 10 7801 GCTTTAAATC TCTGTAGGTA GTTTGTCCAA TTATGTCACA CCACAGAAGT 7851 AAGGTTCCTT CACAAAGATC CGGGACCAAA GCGGCCATCG TGCCTCCCCA 7901 CTCCTGCAGT TCGGGGGCAT GGATGCGCGG ATAGCCGCTG CTGGTTTCCT 7951 GGATGCCGAC GGATTTGCAC TGCCGGTAGA ACTCCGCGAG GTCGTCCAGC 8001 CTCAGGCAGC AGCTGAACCA ACTCGCGAGG GGATCGAGCC CGGGGTGGGC 15 8051 GAAGAACTCC AGCATGAGAT CCCCGCGCTG GAGGATCATC CAGCCGGCGT 8101 CCCGGAAAAC GATTCCGAAG CCCAACCTTT CATAGAAGGC GGCGGTGGAA 8151 TCGAAATCTC GTGATGGCAG GTTGGGCGTC GCTTGGTCGG TCATTTCGAA 8201 CCCCAGAGTC CCGCTCAGAA GAACTCGTCA AGAAGGCGAT AGAAGGCGAT 8251 GCGCTGCGAA TCGGGAGCGG CGATACCGTA AAGCACGAGG AAGCGGTCAG 20 8301 CCCATTCGCC GCCAAGCTCT TCAGCAATAT CACGGGTAGC CAACGCTATG 8351 TCCTGATAGC GGTCCGCCAC ACCCAGCCGG CCACAGTCGA TGAATCCAGA 8401 AAAGCGGCCA TTTTCCACCA TGATATTCGG CAAGCAGGCA TCGCCATGGG 8451 TCACGACGAG ATCCTCGCCG TCGGGCATGC GCGCCTTGAG CCTGGCGAAC 8501 AGTTCGGCTG GCGCGAGCCC CTGATGCTCT TCGTCCAGAT CATCCTGATC 25 8551 GACAAGACCG GCTTCCATCC GAGTACGTGC TCGCTCGATG CGATGTTTCG 8601 CTTGGTGGTC GAATGGGCAG GTAGCCGGAT CAAGCGTATG CAGCCGCCGC 8651 ATTGCATCAG CCATGATGGA TACTTTCTCG GCAGGAGCAA GGTGAGATGA 8701 CAGGAGATCC TGCCCCGGCA CTTCGCCCAA TAGCAGCCAG TCCCTTCCCG 8751 CTTCAGTGAC AACGTCGAGC ACAGCTGCGC AAGGAACGCC CGTCGTGGCC 30 8801 AGCCACGATA GCCGCGCTGC CTCGTCCTGC AGTTCATTCA GGGCACCGGA 8851 CAGGTCGGTC TTGACAAAAA GAACCGGGCG CCCCTGCGCT GACAGCCGGA 8901 ACACGGCGGC ATCAGAGCAG CCGATTGTCT GTTGTGCCCA GTCATAGCCG 8951 AATAGCCTCT CCACCCAAGC GGCCGGAGAA CCTGCGTGCA ATCCATCTTG -57 9001 TTCAATCATG CGAAACGATC CTCATCCTGT CTCTTGATCA GATCTTGATC 9051 CCCTGCGCCA TCAGATCCTT GGCGGCAAGA AAGCCATCCA GTTTACTTTG 9101 CAGGGCTTCC CAACCTTACC AGAGGGCGCC CCAGCTGGCA ATTCCGGTTC 9151 GCTTGCTGTC CATAAAACCG CCCAGTCTAG CTATCGCCAT GTAAGCCCAC 5 9201 TGCAAGCTAC CTGCTTTCTC TTTGCGCrG CGTTTTCCCT TGTCCAGATA 9251 GCCCAGTAGC TGACATTCAT CCGGGGTCAG CACCGTTTCT GCGGACTGGC 9301 TTTCTACGTG TTCCGCTTCC TTTAGCAGCC CTTGCGCCCT GAGTGCTTGC 9351 GGCAGCGTGA AG 10 P OPER\HPM\NovantsUl2827250\Spc Pages doc-14A5/29 - 57A Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. 5 The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of 10 endeavour to which this specification relates.

Claims (17)

1. Use of a humanised antibody that binds CD45RO/RB comprising at least one antigen binding site, comprising hypervariable regions CDRI, CDR2 and CDR3, said CDRI having the amino acid sequence Asn-Tyr-Ile-Ile-His (NYIIH), said CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His-Gly-Thr-Lys-Tyr-Asn-Glu-Lys Phe-Lys-Gly (YFNPYNHGTKYNEKFKG) and said CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-Ala-Trp-Phe-Asp-Thr (SGPYAWFDT) in the manufacture of a medicament.

2. Use according to Claim 1, wherein the humanised antibody that binds CD45RO/RB comprises: a) a first domain comprising the hypervariable regions CDR1, CDR2 and CDR3, said CDR1 having the amino acid sequence Asn-Tyr-Ile-Ile-His (NYIIH), said CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His-Gly-Thr-Lys-Tyr Asn-Glu-Lys-Phe-Lys-Gly (YFNPYNHGTKYNEKFKG) and said CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-Ala-Trp-Phe-Asp-Thr (SGPYAWFDT); and b) a second domain comprising the hypervariable regions CDRl', CDR2' and CDR3', CDRI' having the amino acid sequence Arg-Ala-Ser-Gln-Asn-Ile-Gly-Thr-Ser-Ile Gln (RASQNIGTSIQ), CDR2' having the amino acid sequence Ser-Ser-Ser-Glu-Ser-Ile Ser (SSSESIS) and CDR3' having the amino acid sequence Gln-Gln-Ser-Asn-Thr-Trp-Pro Phe-Thr (QQSNTWPFT).

3. Use according to any one of Claim I or 2, wherein the antibody, comprises a polypeptide of SEQ ID NO: 1 and/or a polypeptide of SEQ ID NO:2.

4. Use according to Claim I or 2, wherein the antibody comprises a polypeptide of SEQ ID NO:3 and/or a polypeptide of SEQ ID NO:4. P:\OPER\HPM\No,,S\12927250\Spe Pags do.14A)5/2009 -59

5. Use of a humanised antibody that binds CD45RO/RB, the antibody comprising a polypeptide of SEQ ID NO:9 or of SEQ ID NO:10 and a polypeptide of SEQ ID NO:7 or of SEQ ID NO:8 in the manufacture of a medicament.

6. Use of a humanised antibody comprising: - a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:7, - a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:8, - a polypeptide of SEQ ID NO:10 and a polypeptide of SEQ ID NO:7, or - a polypeptide of SEQ ID NO:10 and a polypeptide of SEQ ID NO:8 in the manufacture of a medicament.

7. An isolated polynucleotide encoding the humanised antibody of any one of Claims I to 6.

8. An isolated polynucleotide according to Claim 7 encoding the amino acid sequence of CDRI, CDR2 and CDR3 and an isolated polynucleotide encoding the amino acid sequence of CDRI', CDR2' and CDR3'.

9. An isolated polynucleotide of SEQ ID NO:5 and/or a polynucleotide of SEQ ID NO:6.

10. An isolated polynucleotide encoding a polypeptide of SEQ ID NO:7 or SEQ ID NO:8 and a polypeptide of SEQ ID NO:9 or SEQ ID NO:10.

11. An isolated polynucleotide of SEQ ID NO: II or SEQ ID NO: 12 and an isolated polynucleotide of SEQ ID NO:13 or SEQ ID NO:14.

12. An expression system comprising the polynucleotides according to any one of Claims 7 to 11. P OPER\HPM\Novanis\282725(mSpec Paesdoc-14/0512C9 - 60

13. An expression system comprising the polynucleotide according to any one of Claims 7 to 11, wherein said expression system or part thereof is capable of producing an amino acid sequence defined in any one of Claims 1 to 8, when said expression system or part thereof is present in a compatible host cell.

14. An isolated host cell which comprises an expression system according to Claim 12 or 13.

15. Use according to any one of Claims I to 6 in the manufacture of a medicament for treatment and/or prophylaxis of autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies.

16. A method of treatment and/or prophylaxis of diseases associated with autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies comprising administering to a subject in need of such treatment and/or prophylaxis an effective amount of a molecule or a humanised antibody according to any one of Claims 1 to 6.

17. A use of humanised antibody according to any one of Claims I to 6 and 15 or isolated polynucleotides according to any one of Claims 7 to I1 or an expression vector according to Claim 12 or an expression system according to Claim 13 or an isolated host cell according to Claim 14 or a method according to Claim 16 substantially as hereinbefore described with reference to the Figures and Examples.