Human Monoclonal Antibodies Against Human Il-4

Abstract

Antibodies which are specific for human interleukin-4 and their use in the treatment of IL-4 and/or IgE mediated diseases.

Description

[0001] The present invention relates to antibodies which are specific for human interleukin-4 (hIL-4).

[0002] Allergic diseases such as atopic dermatitis, allergic rhinitis, asthma and food allergies are characteristically associated with exacerbated Th2 cell responses to innocuous environmental antigens (allergens). Allergens are captured by antigen presenting cells, processed and presented in the context of MHC Class II molecules to allergen-specific T helper (Th) cells. Allergen specific Th cells belong to the Th2 phenotype and develop from precursor T cells under the influence of interleukin-4 (IL-4). Once Th2 cells are activated, they secrete IL-4 and interleukin-13 (IL-13), which together with surface bound signals induce B cells to switch to IgE producing plasma cells. IgE molecules bind to high affinity Fc.epsilon.IR on mast cells and, after subsequent encounter with allergen, induce mast cell activation and the release of mediators of allergic reactions. Th2 cytokines also promote the survival of eosinophils and the growth of mast cells which, after degranulation, also release additional Th2 cytokines capable of augmenting IgE production, Th2 cell differentiation and eosinophil survival. Thus, Th2 cells play a pivotal role in the induction and development of allergic responses and therefore, antagonizing their development and/or their effector functions would be an efficient way to intervene in allergic responses.

[0003] IL-4 and IL-13 share many biological activities due to the fact that both cytokines use the IL-4 receptor (IL-4R)-alpha chain as a component of their respective receptor complexes. IL-13 signals through an heterodimeric complex consisting of an IL-13 binding chain (1 L-13R.alpha.1) and the IL-4R.alpha. chain. IL-4 utilizes this IL-4R.alpha./IL-13R.alpha.1 complex, called type II IL-4R, as an alternative to the type I IL-4R, consisting of IL-4R.alpha. chain and the common .gamma. chain (c.gamma.) shared by receptors for IL-2, IL-4, IL-7, IL-9, IL-15 and thymic stromal lymphopoietin (TSLP). Because T cells do not express 1 L-13R.alpha.1, IL-13 in contrast to IL-4 does not support T cell proliferation and cannot induce the differentiation of naive human Th cells towards the Th2 phenotype (see e.g. J. E. de Vries et al., Encyclopedia of Hormones and related cell regulators, Academic Press, 2002). IL-4 plays a pivotal role in T cell proliferation and thus in the development and maintenance of allergic diseases. IL-4 gene deficient mice or mice lacking IL-4 (see e.g. Kuhn R. et al., Science, 1991 (5032) 707:10) or the downstream signaling factor STATS (see e.g. Kaplan M. H. et al., Immunity, 1996 (3) 313-9) do not develop significant numbers of Th2 cells and have reduced IgE responses.

[0004] We have now found antibodies with a high affinity for human IL-4 and a strong inhibitory potential of IL-4 mediated IgE synthesis by naive human B cells.

[0005] In one aspect the present invention provides an human IL-4 specific antibody which binds to human IL-4 with a dissociation constant K.sub.d of equal or less than 800 pM, such as e.g. equal or less than 200 pM.

[0006] In another aspect the present invention provides an antibody having a first domain comprising in sequence the hypervariable regions CDR1, CDR2 and CDR3 and a second domain comprising in sequence the hypervariable regions CDR1', CDR2' and CDR3' selected from the group consisting of an antibody wherein [0007] a) said CDR1 has the amino acid sequence Gly-Phe-Thr-Phe-Ser-Ser-Tyr-Ala-Met-His (GFTFSSYAMH) (SEQ ID NO: 49), [0008] said CDR2 has the amino acid sequence Phe-Ile-Trp-Asp-Asp-Gly-Ser-Phe-Lys-Tyr-Tyr-Ala-Glu-Ser-Val-Lys-Gly (FIWDDGSFKYYAESVKG) (SEQ ID NO: 50), [0009] said CDR3 has the amino acid sequence Glu-Gly-Ser-Trp-Ser-Pro-Asp-Ile-Phe (EGSWSPDIF) (SEQ ID NO: 51), [0010] said CDR1' has the amino acid sequence Ser-Gln-Gly-Ile-Ser-Arg-Ala (SQGISRA) (SEQ ID NO: 52), [0011] said CDR2' has the amino acid sequence Asp-Ala-Ser (DAS) (SEQ ID NO: 53), [0012] said CDR3' has the amino acid sequence Phe-Asn-Ser-Tyr-Pro-Ile (FNSYPI) (SEQ ID NO: 54), [0013] b) said CDR1 has the amino acid sequence Gly-Phe-Thr-Leu-Ser-Ser-Phe-Gly-Met-His (GFTLSSFGMH) (SEQ ID NO: 55), [0014] said CDR2 has the amino acid sequence Val-Ile-Trp-Tyr-Asp-Gly-Ser-Asn-Glu-Tyr-Tyr-Ala-Asp-Ser-Val-Lys-Gly (VIWYDGSNEYYADSVKG) (SEQ ID NO: 56), [0015] said CDR3 has the amino acid sequence Glu-Gly-Ser-Trp-Ser-Pro-Asp-Ile-Phe (EGSWSPDIF) (SEQ ID NO: 57), [0016] said CDR1' has the amino acid sequence Ser-Gln-Gly-Ile-Arg-Ser-Ala (SQGIRSA) (SEQ ID NO: 58), [0017] said CDR2' has the amino acid sequence Asp-Ala-Ser (DAS) (SEQ ID NO: 53), [0018] said CDR3' has the amino acid sequence Phe-Asn-Ser-Tyr-Pro-Val (FNSYPV) (SEQ ID NO: 59), [0019] c) said CDR1 has the amino acid sequence Gly-Phe-Thr-Leu-Ser-Ser-Tyr-Gly-Met-His (GFTLSSYGMH) (SEQ ID NO: 60), [0020] said CDR2 has the amino acid sequence Val-Ile-Trp-Tyr-Asp-Gly-Asn-Asn-Gln-Tyr-Tyr-Ala-Asp-Ser-Val-Lys-Gly (VIWYDGNNQYYADSVKG) (SEQ ID NO: 61), [0021] said CDR3 has the amino acid sequence Glu-Gly-Ser-Trp-Ser-Pro-Asp-Ile-Phe (EGSWSPDIF) (SEQ ID NO: 62), [0022] said CDR1' has the amino acid sequence Ser-Gln-Gly-Ile-Ser-Ser-Tyr (SQGISSY) (SEQ ID NO: 63), said CDR2' has the amino acid sequence Asp-Ala-Ser (DAS) (SEQ ID NO: 53), [0023] said CDR3' has the amino acid sequence Phe-Asn-Ser-Tyr-Pro (FNSYP) (SEQ ID NO: 64), [0024] d) said CDR1 has the amino acid sequence Gly-Asp-Thr-Phe-Ser-Ser-Tyr-Ala-Ile-Ser (GDTFSSYAIS) (SEQ ID NO: 65), [0025] said CDR2 has the amino acid sequence Gly-Ile-Ile-Pro-Val-Ile-Gly-Thr-Val-Asn-Tyr-Glu-Glu-Arg-Phe-Gln-- Asp (GIIPVIGTVNYEERFQD) (SEQ ID NO: 66), [0026] said CDR3 has the amino acid sequence Glu-Glu-Gly-Phe-Leu (EEGFL) (SEQ ID NO: 67), [0027] said CDR1' has the amino acid sequence Ser-Gln-Gly-Ile-Ser-Ser-Ala (SQGISSA) (SEQ ID NO: 68), [0028] said CDR2' has the amino acid sequence Asp-Ala-Ser (DAS) (SEQ ID NO: 53), [0029] said CDR3' has the amino acid sequence Phe-Asn-Ser-Tyr-Pro-Leu (FNSYPL) (SEQ ID NO: 69), [0030] e) said CDR1 has the amino acid sequence Gly-Phe-Thr-Phe-Ser-Cys-Cys-Gly-Met-His (GFTFSCCGMH) (SEQ ID NO: 70), [0031] said CDR2 has the amino acid sequence Val-Ile-Trp-Tyr-Asp-Gly-Ser-Asn-Lys-Tyr-Tyr-Ala-Asp-Ser-Val-Lys-Gly (VIWYDGSNKYYADSVKG) (SEQ ID NO: 71), [0032] said CDR3 has the amino acid sequence Asp-Ser-Ser-Gly-Ser-Phe-Tyr-Glu-Tyr-Phe (DSSGSFYEYF) (SEQ ID NO: 72), [0033] said CDR1' has the amino acid sequence Ser-Gln-Gly-Ile-Asn-Ser-Ala (SQGINSA) (SEQ ID NO: 73, [0034] said CDR2' has the amino acid sequence Asp-Ala-Ser (DAS) (SEQ ID NO: 53), [0035] said CDR3' has the amino acid sequence Phe-Asn-Ser-Tyr-Pro-Tyr (FNSYPY) (SEQ ID NO: 74), and [0036] f) said CDR1 has the amino acid sequence Gly-Phe-Thr-Phe-Ser-Gly-Tyr-Gly-Met-His (GFTFSGYGMH) (SEQ ID NO: 75), [0037] said CDR2 has the amino acid sequence Val-Val-Trp-Tyr-Asp-Gly-Gly-Tyr-Lys-Phe-Tyr-Ala-Asp-Ser-Val-Lys-Gly (VVWYDGGYKFYADSVKG) (SEQ ID NO: 76), [0038] said CDR3 has the amino acid sequence Asp-Ser-Ser-Gly-Ser-Phe-Tyr-Glu-Tyr-Leu (DSSGSFYEYL) (SEQ ID NO: 77), [0039] said CDR1' has the amino acid sequence Ser-Gln-Gly-Ile-Ser-Ser-Ala (SQGISSA) (SEQ ID NO: 78), [0040] said CDR2' has the amino acid sequence Asp-Ala-Ser (DAS) (SEQ ID NO: 53), [0041] said CDR3' has the amino acid sequence Phe-Asn-Ser-Tyr-Pro-His (FNSYPH) (SEQ ID NO: 79).

[0042] CDR1, CDR2 and CDR3 are part of the amino acid sequence of the heavy chain of such antibody and CDRI', CDR2' and CDR3' are part of the amino acid sequence of the light chain of such antibody.

[0043] We also have found an antibody comprising: [0044] the amino acid sequence of the heavy chain of a mature polypeptide of SEQ ID NO:1 and the amino acid sequence of the light chain of a mature polypeptide of SEQ ID NO:2, or [0045] the amino acid sequence of the heavy chain of a mature polypeptide of SEQ ID NO:9 and the amino acid sequence of the light chain of a mature polypeptide of SEQ ID NO:10, or [0046] the amino acid sequence of the heavy chain of a mature polypeptide of SEQ ID NO:17 and the amino acid sequence of the light chain of a mature polypeptide of SEQ ID NO:18, or [0047] the amino acid sequence of the heavy chain of a mature polypeptide of SEQ ID NO:25 and the amino acid sequence of the light chain of a mature polypeptide of SEQ ID NO:26, or [0048] the amino acid sequence of the heavy chain of a mature polypeptide of SEQ ID NO:33 and the amino acid sequence of the light chain of a mature polypeptide of SEQ ID NO:34, or [0049] the amino acid sequence of the heavy chain of a mature polypeptide of SEQ ID NO:41 and the amino acid sequence of the light chain of a mature polypeptide of SEQ ID NO:42.

[0050] In a further aspect the present invention provides an antibody comprising

a) a polypeptide of SEQ ID NO:1 and a polypeptide of SEQ ID NO:2, or b) a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:10, or c) a polypeptide of SEQ ID NO: 17 and a polypeptide of SEQ ID NO: 18, or d) a polypeptide of SEQ ID NO:25 and a polypeptide of SEQ ID NO:26, or e) a polypeptide of SEQ ID NO:33 and a polypeptide of SEQ ID NO:34, or f) a polypeptide of SEQ ID NO:41 and a polypeptide of SEQ ID NO:42.

[0051] We further have found an antibody comprising: [0052] the amino acid sequence of the heavy chain of a polypeptide of SEQ ID NO:1 further containing a leader sequence (=SEQ ID NO:3) and the amino acid sequence of the light chain of a polypeptide of SEQ ID NO:2 further containing a leader sequence (=SEQ ID NO:4), or [0053] the amino acid sequence of the heavy chain of a polypeptide of SEQ ID NO:9 further containing a leader sequence (=SEQ ID NO:11) and the amino acid sequence of the light chain of a polypeptide of SEQ ID NO:10 further containing a leader sequence (=SEQ ID NO:12), or [0054] the amino acid sequence of the heavy chain of a polypeptide of SEQ ID NO:17 further containing a leader sequence (=SEQ ID NO:19) and the amino acid sequence of the light chain of a polypeptide of SEQ ID NO:18 further containing a leader sequence (=SEQ ID NO:20), or [0055] the amino acid sequence of the heavy chain of a polypeptide of SEQ ID NO:25 further containing a leader sequence (=SEQ ID NO:27) and the amino acid sequence of the light chain of a polypeptide of SEQ ID NO:26 further containing a leader sequence (=SEQ ID NO:28), or [0056] the amino acid sequence of the heavy chain of a polypeptide of SEQ ID NO:33 further containing a leader sequence (=SEQ ID NO:35) and the amino acid sequence of the light chain of a polypeptide of SEQ ID NO:34 further containing a leader sequence (=SEQ ID NO:36), or [0057] the amino acid sequence of the heavy chain of a polypeptide of SEQ ID NO:41 further containing a leader sequence (=SEQ ID NO:43) and the amino acid sequence of the light chain of a polypeptide of SEQ ID NO:42 further containing a leader sequence (=SEQ ID NO:44).

[0058] In a further aspect the present invention provides an antibody comprising

a) a polypeptide of SEQ ID NO:3 and a polypeptide of SEQ ID NO:4, or b) a polypeptide of SEQ ID NO:11 and a polypeptide of SEQ ID NO:12, or c) a polypeptide of SEQ ID NO:19 and a polypeptide of SEQ ID NO:20, or d) a polypeptide of SEQ ID NO:27 and a polypeptide of SEQ ID NO:28, or e) a polypeptide of SEQ ID NO:35 and a polypeptide of SEQ ID NO:36, or f) a polypeptide of SEQ ID NO:43 and a polypeptide of SEQ ID NO:44.

[0059] Antibodies provided by the present invention are hereinafter also designated as "compound(s) of (according to) the present invention".

[0060] In another aspect the present invention provides a compound of the present invention which is selected from the group consisting of an human IL-4 specific monoclonal antibody (hIL-4 mAb), a fragment thereof and an analog thereof.

[0061] An hIL-4 mAb is an antibody which specifically recognizes human IL-4, i.e. includes antigen binding sites specific for human IL-4, and which has specifically its CDRs but also other parts of the heavy and light chain derived from human immunoglobulins. The antibody may be of any isotype including IgG1, IgG2, IgG3 and IgG4, preferably of isotype IgG1.

[0062] "A fragment thereof" means a part of the heavy and light chain variable sequence of a hIL-4 mAb, which retains the same antigen binding specificity and/or neutralizing ability as the molecule from which the fragments are derived, e.g. a Fab fragment or a F(ab').sub.2 fragment derived from hIL-4 mAb. A Fab fragment contains the entire light chain and amino terminal portions of the heavy chain; a F(ab').sub.2 fragment is the fragment formed by 2 Fab fragments bound by disulfide bonds. Such fragments can be obtained by conventional means, e.g. cleavage of the monoclonal antibodies with the appropriate proteolytic enzymes, papain and/or pepsin, or by recombinant methods, and the fragments themselves are useful as therapeutic and/or prophylactic agents.

[0063] "An analog thereof" means a hIL-4 mAb with an amino acid sequence which is modified by at least one amino acid outside of the CDR regions, e.g. outside of CDR1, CDR2 and CDR3 of the heavy chain or outside of CDR1', CDR2' and CDR3' of the light chain. Said modification includes a chemical modification, a substitution or a rearrangement of one or a few amino acids, i.e. no more than 10 amino acids, which modification permits the amino acid sequence to retain the biological characteristics, e.g. antigen specificity and affinity, of the unmodified sequence. For example silent mutations can be constructed via substitution to create endonuclease restriction sites within or surrounding the CDR regions.

[0064] An analog may also arise as allelic variation. An "allelic variation or modification" is an alteration in the nucleic acid sequence encoding an antibody of the present invention outside of the CDR regions. Such alterations or modifications may be due to the degeneracies of the genetic code or may be liberately engineered to provide desired characteristics. Such variations or modifications may or may not result in alterations in any encoded amino acid sequence, but retain the biological activities, e.g. antigen specificity and affinity.

[0065] We have also found polynucleotides encoding compounds of the present invention.

[0066] In another aspect the present invention provides isolated polynucleotides comprising polynucleotides encoding a compound of the present invention.

[0067] In another aspect the present invention provides polynucleotides encoding the amino acid sequence of CDR1, CDR2 and CDR3 of a compound of the present invention and polynucleotides encoding the amino acid sequence of CDR1', CDR2' and CDR3' of a compound of the present invention.

[0068] In another aspect the present invention provides polynucleotides comprising

a) a polynucleotide of SEQ ID NO:5 and a polynucleotide of SEQ ID NO:6, or b) a polynucleotide of SEQ ID NO:13 and a polynucleotide of SEQ ID NO:14, or c) a polynucleotide of SEQ ID NO:21 and a polynucleotide of SEQ ID NO:22, or d) a polynucleotide of SEQ ID NO:29 and a polynucleotide of SEQ ID NO:30, or e) a polynucleotide of SEQ ID NO:37 and a polynucleotide of SEQ ID NO:38, or f) a polynucleotide of SEQ ID NO:45 and a polynucleotide of SEQ ID NO:46.

[0069] In another aspect the present invention provides polynucleotides encoding

a) a polypeptide of SEQ ID NO:7 and SEQ ID NO:8, or b) a polypeptide of SEQ ID NO:15 and SEQ ID NO:16, or c) a polypeptide of SEQ ID NO:23 and SEQ ID NO:24, or d) a polypeptide of SEQ ID NO:31 and SEQ ID NO:32, or e) a polypeptide of SEQ ID NO:39 and SEQ ID NO:40, or f) a polypeptide of SEQ ID NO:47 and SEQ ID NO:48. SEQ ID NO:5 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:1. SEQ ID NO:6 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:2. SEQ ID NO:7 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:3. SEQ ID NO:8 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:4. SEQ ID NO:13 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:9. SEQ ID NO:14 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:10. SEQ ID NO:15 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:11. SEQ ID NO:16 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:12. SEQ ID NO:21 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:17: SEQ ID NO:22 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:18. SEQ ID NO:23 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:19. SEQ ID NO:24 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:20. SEQ ID NO:29 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:25. SEQ ID NO:30 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:26. SEQ ID NO:31 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:27. SEQ ID NO:32 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:28. SEQ ID NO:37 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:33. SEQ ID NO:38 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:34. SEQ ID NO:39 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:35. SEQ ID NO:40 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:36. SEQ ID NO:45 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:41. SEQ ID NO:46 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:42. SEQ ID NO:47 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:43. SEQ ID NO:48 is a polynucleotide encoding an amino acid sequence of SEQ ID NO:44.

[0070] A compound of the present invention may be produced by recombinant DNA techniques. Thus, one or more DNA molecules encoding the antibody, a fragment thereof or an analog thereof may be constructed, placed under appropriate control sequences in an appropriate vector and transferred into a suitable host (organism) for expression. The compound of the present invention 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 hypervariable and/or variable regions and the polynucleotides 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 replicable expression vector including a suitable promoter operably linked to a polynucleotide sequence of interest, e.g. encoding at least a variable domain of an immunoglobulin heavy or light chain comprising CDRs, is prepared, a suitable cell line is transformed with said expression vector, the transformed cell line is cultured and the corresponding immunoglobulin is obtained.

[0071] In another aspect the present invention provides an expression vector comprising a polynucleotide encoding a compound of the present invention, e.g. at least one polynucleotide of SEQ ID NO:5, SEQ ID NO: 6, SEQ ID NO:7, SEQ ID No:8, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID No:16, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID No:24, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID No:40, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47 or SEQ ID No:48.

[0072] Naturally an expression vector can comprise more than one polynucleotide.

[0073] In another aspect the present invention provides [0074] An expression system comprising a polynucleotide encoding a compound of the present invention wherein said expression system or part thereof is capable of producing a compound of the present invention, e.g. a hIL-4 mAb, when said expression system or part thereof is present in a compatible host cell; and [0075] An isolated host cell comprising an expression system as defined above.

[0076] Expression vectors, e.g. comprising suitable promoter(s) and genes encoding heavy and light chain constant parts are known, e.g. and are commercially available and include e.g. heavy chain vector IgG1 and light chain vector human kappa. A conventional expression vector or recombinant plasmid may be produced by placing the respective polynucleotide in operative association with conventional regulatory control sequences capable of controlling the replication and expression in, and/or secretion from, a host cell. Regulatory sequences include promoter sequences, e.g. CMV promoter, LCK promoter, and appropriate signal sequences.

[0077] A selected host cell may be transfected by conventional techniques with the vector of interest to create a transfected host cell, which then may be cultured by conventional techniques to produce the compounds of the present invention. Appropriate cell lines may be found according, e.g. analogously, to a method as conventional. Appropriate hosts are known or may be found according, e.g. analogously, to a method as conventional and include cell cultures or transgenic animals. Suitable host cells or cell lines for the expression of the compounds of the present invention are preferably eukaryotic cells such as e.g. CHO, COS, a fibroblast cell (e.g. 3T3) and myeloid cells among others, preferably a mammalian cell, such as a CHO cell or SP2/0.

[0078] The compounds of the present invention exhibit pharmacological activity and are therefore useful as pharmaceuticals. E.g., the compounds of the present invention interfere strongly with the binding of IL-4 to an IL-4 receptor and are herein also referred to as IL-4 blocking or neutralizing compounds of the present invention, including hIL-4 mAb(s). Compounds of the present invention show activity and their affinity can be determined in the TEST: AFFINITY MEASUREMENT as described in Example 1.

[0079] A compound of the present invention thus shows therapeutic activity against IL-4 and/or IgE mediated diseases, such as [0080] various allergic diseases, e.g. including urticaria, allergic reactions to medication, rhinitis, e.g. allergic rhinitis, conjunctivitis, e.g. rhinoconjunctivitis, dermatitis, e.g. atopic dermatitis, asthma, e.g. atopic asthma and allergic asthma, anaphylactic shock; preferably atopic dermatitis, allergic asthma, allergic rhinitis, allergic rhinoconjunctivitis, such as allergic asthma or atopic dermatitis; [0081] autoimmune diseases, including e.g. Kawasaki disease, Grave's disease, Sjorgen's syndrome, autoimmune lymphoproliferative syndrome, autoimmune haemolytic anemia, autoimmune uveitis, myasthenia gravis, Lupus Erythematosis and Bullous pemphigoid; [0082] disorders of the digestive system in which IL-4 and/or IgE play a role, including e.g. ulcers, gastric inflammation, mucosal inflammation, ulcerative colitis, Crohn's disease, inflammatory bowel disease and other disorders of the digestive system in which IL-4 and/or IgE play a role; [0083] diseases wherein IL-4 and/or IgE are overproduced and considered to contribute to pathology, including e.g. systemic sclerosis (scleroderma), septic arthritis and reactive arthritis.

[0084] In another aspect the present invention provides a compound of the present invention, e.g. a hIL-4 mAb, for use as a pharmaceutical, e.g. against IL-4 and/or IgE mediated diseases, e.g. allergic diseases, e.g. atopic dermatitis, allergic asthma, allergic rhinitis, preferably atopic dermatitis.

[0085] In another aspect the present invention provides the use of the present invention of a compound of the present invention which is selected from the group consisting of a hIL-4 mAb, a fragment thereof and an analog thereof.

[0086] For pharmaceutical use a compound of the present invention includes one or more, preferably one, compounds of the present invention, e.g. a combination of two or more compounds of the present invention.

[0087] In another aspect the present invention provides the use of a compound of the present invention for the manufacture of a medicament, e.g. a pharmaceutical composition, for the treatment of diseases mediated by IL-4 and/or IgE, e.g. allergic diseases, e.g. allergic rhinoconjunctivitis, atopic dermatitis, allergic asthma, allergic rhinitis, preferably allergic asthma or atopic dermatitis, such as atopic dermatitis.

[0088] In a further aspect the present invention provides the use of a compound of the present invention for the manufacture of a medicament, e.g. a pharmaceutical composition, for the treatment of a disease as described above, e.g. selected from the group consisting of atopic dermatitis, allergic asthma and allergic rhinitis.

[0089] In a further aspect the present invention provides a compound of the present invention for the uses as mentioned above, wherein the compound of the present invention is selected from the group consisting of a hIL-4 mAb, a fragment thereof and an analog thereof.

[0090] It has, for example been determined that the affinity constant of a compound of the present invention, e.g. a hIL-4 mAb, a fragment thereof or an analog thereof, for human IL-4 is equal or less than 800 pM, e.g. is equal or less than 200 pM, such as of about 30 pM to about 200 pM, preferably of about 45 pM to about 170 pM, such as about 100 pM, more preferred about 50 pM, such as about 45 pM.

[0091] It has, for example also been determined that the affinity constant of a compound of the present invention, e.g. a hIL-4 mAb, a fragment thereof or an analog thereof, for human IL-4 is of 30 pM to 200 pM, preferably of 45 pM to 170 pM, such as 100 pM, more preferred 50 pM, such as 45 pM. It is therefore, indicated that for the treatment of diseases mediated by IL-4, the compounds of the present invention may be administered to larger mammals, for example humans, by similar modes of administration at similar dosages than conventionally used with monoclonal antibodies.

[0092] In another aspect the present invention provides an antibody which binds to human IL-4 with a dissociation constant of equal or less than 200 pM, e.g. 30 to 200 pM.

[0093] In a further aspect of the present invention the antibody for the uses as mentioned above is a hIL-4 mAb, a fragment thereof or an analog thereof which binds to human IL-4 with a dissociation constant K.sub.d of equal or less than 200 pM, e.g. 30 to 200 pM.

[0094] In a further aspect the present invention provides a method of treatment of diseases which are mediated by IL-4 and/or IgE, e.g. allergic diseases, e.g. atopic dermatitis, allergic asthma, allergic rhinitis, preferably atopic dermatitis, which treatment comprises administering to a subject in need of such treatment an effective amount of a compound of the present invention; e.g. in the form of a pharmaceutical composition.

[0095] In a further aspect of the present invention a compound of the present invention is administered in combination with another pharmaceutically active agent either simultaneously or in sequence.

[0096] Treatment includes treatment and prophylaxis.

[0097] For such treatment, the appropriate dosage will, of course, vary depending upon, for example, the chemical nature and the pharmacokinetic data of an antibody of the present invention employed, the individual host, the mode of administration and the nature and severity of the conditions being treated. However, in general, for satisfactory results in larger mammals, for example humans, an indicated daily dosage is in the range from, e.g. about, 0.1 ng/kg to, e.g. about, 10 mg/kg, such as from, e.g. about, 100 ng/kg to, e.g. about, 2 mg/kg of a compound of the present invention; conveniently administered, for example, in divided doses up to four times a day. A compound of the present invention may be administered by any conventional route, for example parenterally, e.g. including intravenous, intradermal, intramuscular, subcutaneous, intranasal administration, injectable solutions or suspensions or inhaler powder.

[0098] In another aspect the present invention provides a pharmaceutical composition comprising a compound of the present invention in association with at least one pharmaceutical excipient, e.g. appropriate carrier and/or diluent, e.g. including fillers, binders, disintegrators, flow conditioners, lubricants, sugars and sweeteners, fragrances, preservatives, stabilizers, wetting agents and/or emulsifiers, solubilizers, salts for regulating osmotic pressure and/or buffers.

[0099] In another aspect the present invention provides a pharmaceutical composition according to the present invention further comprising another pharmaceutically active agent.

[0100] Such compositions may be manufactured according, e.g. analogously to a method as conventional, e.g. by mixing, granulating, coating, dissolving or lyophilizing processes. Unit dosage forms may contain, for example, from, e.g. about, 0.5 mg to, e.g. about, 1000 mg, such as 1 mg to about 500 mg.

[0101] A compound of the present invention may be used for pharmaceutical treatment according to the present invention alone or in combination with one or more other pharmaceutically active agents. Such other pharmaceutically active agents include e.g. other antibodies, e.g. such as antibodies neutralizing IgE, cytokines or cytokine receptors, which are chosen according to the particular condition to be treated.

[0102] Combinations include fixed combinations, in which two or more pharmaceutically active agents are in the same formulation; kits, in which two or more pharmaceutically active agents in separate formulations are sold in the same package, e.g. with instruction for co-administration; and free combinations in which the pharmaceutically active agents are packaged separately, but instruction for simultaneous or sequential administration are given.

[0103] In another aspect the present invention also provides a method for diagnosing allergies and other conditions associated with excess IgE production in a human which comprises

a) contacting a sample of a biological fluid with an antibody of the present invention, and b) assaying for the occurrence of binding between said antibody and human IL-4.

[0104] In the following Examples all temperatures are in degrees Celsius (.degree. C.) and are uncorrected.

[0105] The following abbreviations are used:

FACS Buffer PBS, 2% FCS and 0.2% NaN.sub.3

[0106] FCS fetal calf serum FITC fluoroescein isothiocyanat IC.sub.50 inhibitory concentration KLH keyhole limpet hemocyanin mAb monoclonal antibody MTX methotrexate PBS phosphate buffered saline slgD surface Immunoglobulin D rmIL-3 recombinant murine interleukin-3 rhIL-4 recombinant human interleukin-4 rhIL-13 recombinant human interleukin-13 rpm revolutions per minute RPMI Roswell Park Memorial Institute medium RT room temperature TF-buffer 272 mM sucrose, 1 mM MgCl.sub.2, 7 mM phosphate buffer pH 7.4

EXAMPLES

Example 1

a) Antibody Production

[0107] Plasmids encoding the variable regions of the sequence SEQ ID NO: 5 or 7 (=heavy chain) and the sequence SEQ ID NO: 6 or 8 (=light chain) of the antibody are cloned into expression cassettes for human kappa light chains and human IgG1 heavy chains.

[0108] The specificity determining regions are combined with the necessary elements to generate complete monoclonal antibodies, i.e. promoter, leader sequence and splice donor sites for splicing to the antibody constant region exons that are required for the expression of functional immunoglobulin proteins. The variable region cassettes for the heavy and light chain antibodies, encoding for leader sequences, variable region and 3'-prime splice donor sites for splicing the constant region exons CH1-CH4 and kappa, are transferred into mammalian expression vectors HC (heavy chain vector, human IgG1) and LC (light chain vector, human kappa).

[0109] The light chain containing plasmid and the heavy chain containing plasmid are introduced into Sp2/0 cells in a co-transfection approach. E.g. for transfection, cells in exponential growth phase with a viability of about 95% are used. Cells are washed twice with cold TF-buffer and cell concentration is adjusted to 2.times.10.sup.7 cells/ml in TF-buffer. 0.8 ml cell suspension obtained are mixed with 15 .mu.g each of the heavy chain and light chain plasmid and placed on ice for 10 minutes. Transfection is done by electroporation using the Biorad Gene Pulser (280 V and 25 .mu.F). After electroporation, cells are placed on ice for 15 minutes, transferred into 50 ml cold culture medium and incubated for 1 day at 37.degree. and 5% CO.sub.2. For clonal amplification, the G418 resistant cells obtained are cultivated in the presence of 200 nM MTX. An aliquot of the heterogeneous cell pool is seeded into 96-well plates at clonal density of 1 viable cell/well in culture medium containing 200 nM MTX allowing selection of clonal populations of amplified cells. Limiting dilution cloning is applied to generate clonal cell lines after amplification and adaptation to serum-free culture conditions. Cells are seeded into two 96-well plates at a concentration of 0.5 cell/well. Wells are screened microscopically for clonality one day after seeding. Only monoclonal antibodies are used for further testing.

[0110] The antibody obtained comprises an amino acid sequence of SEQ ID NO:1 and SEQ ID NO:2.

b) Affinity Measurements

[0111] Affinity measurements of human mAbs of the present invention are carried on a BIAcore.TM. 2000 instrument. Anti-human IgG is coated onto a BIAcore sensorchip CM-5 (BIAcore), so that application of defined amounts of human mAbs results in capturing on this prepared surface and hence in a change of refractory properties that are measured. A subsequent application of rhIL-4 results in a further change of the refractory properties, which allows determination of the association rate (K.sub.on, on rate) as well as the dissociation rate (K.sub.off, off rate) and the product of these two, the affinity (K.sub.d, dissociation constant, given in pM), according to the BIAevaluation 3.0 software. Using several concentrations of rhIL-4 in BIAbuffer, an affinity of 43 pM of an mAb as defined by CDRs as described in claim 2a) is determined.

c) Determination of Inhibitory Potential on IL-4 Mediated IgE

[0112] c1) Cell Sources

[0113] Human B cells are isolated from peripheral blood, respectively, buffy coats by Ficoll-paque density centrifugation, followed by magnetic separation with MACS beads (Miltenyi Biotech) specific for human CD19 or human CD22 on an AutoMACS device.

[0114] Naive human B cells are similarly isolated using anti-human slgD FITC labeled goat F(ab').sub.2 antibodies followed by anti-FITC MACS beads.

c2) Cell Culture/Maintenance

[0115] Transfectants (BaF/3 transfectants carrying the IL-4R.alpha. and the IL-13R.alpha.1) are cultured in RPMI 1640 medium supplemented with Glutamax (Invitrogen), 10% FCS, 1% penicillin/streptomycin and 10 ng/ml rhIL-4 (Novartis). Cells are split 1:1 twice weekly, washed with fresh medium without rhIL-4 and kept in such medium overnight (=starved cells).

[0116] Human ex vivo (naive) B cell cultures are incubated in X-Vivo medium (Cambrex, XV15) supplemented with Glutamax, 10% FCS, 1% penicillin/streptomycin in 96-well plates (Costar).

c3) BAF Cell Proliferation Assay

[0117] Starved cells as described in b) are collected, washed with fresh medium, counted and adjusted to 2.times.10.sup.5 cells/ml of which 100 .mu.l per well are distributed to 96-well plates (Costar). For titration series the cytokines rhIL-4, rhIL-13 and rmIL-3 are prepared in 4 times the desired final concentration in the same medium.

[0118] For titration series antibodies are either used as cell culture supernatants with ELISA determined concentrations or from purified material at 4 times the desired concentrations in the same medium. The pre-dilutions of cytokines and antibodies are mixed at equal volumes and 100 .mu.l of the pre-mix are transferred to the well prepared with cells. Controls are set up for background proliferation (medium without cytokine and without antibody=100% inhibition) and maximum proliferation (medium with cytokine only=0% inhibition).

[0119] After overnight incubation at 37.degree. in the presence of 5% CO.sub.2, [methyl-.sup.3H]thimidine (Amersham TRK120) 1 .mu.Ci per well is added in 10 .mu.l of medium and incubated for 8 hours. Following a freeze/thaw cycle cells are harvested from the plates on filter mats using a Tomtek harvester. Filter mats are dried in a microwave oven for 2 minutes at 650 W and transferred to a sample bag together with a sheet of Meltilex Scintillation Wax (Wallac). Wax is melted through the filter and filters obtained are placed inside appropriate cassettes and inserted into a micro-beta reader (Wallac) for scintillation counting using a program measuring 30 seconds per field and extrapolating to counts per minutes. An IC.sub.50 of 30 pM is measured in this test system for the antibody.

c4) IL-4 Induced CD23 Up-Regulation on B Cells

[0120] MACS separated B cells are adjusted to 0.5-1.times.10.sup.6 cells/ml in XV15 and plated out in 100 .mu.l per well of 96 well round bottom plates. Cytokines (1 ng/ml final) and mAb (2 .mu.g/ml-2 ng/ml final) are pre-diluted and pre-mixed as described above and added in 100 .mu.l to reach a final volume of 200 .mu.l.

[0121] After culturing overnight at 37.degree. in the presence of 5% CO.sub.2 cells are transferred to 96-well plates (Costar) and centrifuged at 2200 rpm (.about.1000.times.g) for 1 minute after flicking off the supernatant washed with FACS buffer.

[0122] Florochrome labeled mAbs [HLA-DR FITC (Caltag #MHLDRO1 1:800), CD19 PE (Caltag #MHCD1904 1:200) and CD23 APC (Caltag #MHCD2305 1:200)] are prepared in FACS buffer and distributed in 50 .mu.l per well. After 30-60 minutes incubation at RT, wells are filled up with FACS buffer, centrifuged and the centrifugation residue obtained is washed with FACS buffer. Cells are re-suspended in FACS buffer with 2 .mu.g/ml propidium iodide and analyzed on a dual laser FacsCalibur flow cytometer (BD Biosciences). Cells are gated according to their forward scatter and side scatter properties as well as their ability to exclude propidium iodide and their CD19 expression. Mean fluorescence intensities and percentage of cells above arbitrary threshold (set on un-induced cells) in CD23 expression are determined. Baseline expression (100% inhibition) is determined on cells without cytokine, whereas 0% inhibition is set on cells incubated with cytokine but without mAb.

[0123] An IC.sub.50 of 334 pM for this antibody is determined for IL-4 induced CD23 expression on the cells as described above in the presence of 70 pM recombinant human IL-4.

c5) IL-4 Induced IgE Production by Naive B Cells

[0124] Magnetically sorted naive B cells are adjusted to 3.times.10.sup.5 cells per ml in XV15 and plated out in 100 .mu.l per well of 96-well plates in a 6.times.6 array in the center of the plate, surrounded by PBS filled wells during the 10 days of culture at 37.degree. in the presence of 5% CO.sub.2. One plate each is prepared per mAb to be tested, consisting of 3 wells each un-induced and induced controls and quintuplicate repeats of mAb titrations starting at 7 .mu.g/ml and running in 3-fold dilution down to 29 ng/ml final concentrations added in 50 .mu.l four times concentrated pre-dilution. Inducing conditions are rhIL-4 at 20 ng/ml plus anti-CD40 mAb (Novartis) at 0.5 .mu.g/ml final concentrations also added in 50 .mu.l of four times concentrated pre-dilution. IgE concentrations are determined at the end of the culture period by a standard sandwich ELISA method.

[0125] An 1050 of 2806 pM for this antibody is determined for IL-4 induced IgE on the cells as described above.

Examples 2 to 6

[0126] Antibodies are obtained analogously as described in example 1 and comprise the following amino acid sequences:

Example 2 is an antibody comprising amino acid sequence SEQ ID NO:9 and SEQ ID NO:10. Example 3 is an antibody comprising amino acid sequence SEQ ID NO:17 and SEQ ID NO:18. Example 4 is an antibody comprising amino acid sequence SEQ ID NO:25 and SEQ ID NO:26. Example 5 is an antibody comprising amino acid sequence SEQ ID NO:33 and SEQ ID NO:34. Example 6 is an antibody comprising amino acid sequence SEQ ID NO:41 and SEQ ID NO:42.

[0127] Table 1 summarizes the IC.sub.50 values of the antibodies in the various test systems as described in Example 1b to c.

TABLE-US-00001 TABLE 1 IC50 [pM] measured in the particular assays BAF cell K.sub.d [pM] proliferation IL-4 induced CD23 IL-4 induced IgE Ex. 1 43 30 334 2806 Ex. 2 59 50 463 5339 Ex. 3 89 84 873 7813 Ex. 4 66 688 848 7570 Ex. 5 84 218 2835 9944 Ex. 6 170 167 2869 2648

SEQUENCE LISTING

TABLE-US-00002 [0128] SEQ ID NO: 1 (amino acid sequence of the mature polypeptide of the heavy chain, underlined amino acids correspond to CDR1, CDR2 and CDR3) QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGLEWVTF IWDDGSFKYYAESVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREG SWSPDIFDIWGQGTMVTVSS SEQ ID NO: 2 (amino acid sequence of the mature polypeptide of the light chain, underlined amino acids correspond to CDR1, CDR2' and CDR3) AIQLTQSPSSLSASVGDRVTITCRASQGISRALAWYQQKPGKAPKLLIYD ASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPITFGQ GTRLEIKRT SEQ ID NO: 3 (amino acid sequence of the polypeptide of SEQ ID NO: 1 with leader sequence, amino acids marked in bold correspond to the leader sequence) MEFGLNWVFLVALFRGVHCQVQLVESGGGVVQPGRSLRLSCAASGFTFSS YAMHWVRQAPGKGLEWVTFIWDDGSFKYYAESVKGRFTISRDNSKNTLYL QMNSLRAEDTAVYYCAREGSWSPDIFDIWGQGTMVTVSS SEQ ID NO: 4 (amino acid sequence of the polypeptide of SEQ ID NO: 2 with leader sequence, amino acids marked in bold correspond to the leader sequence) MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRASQG ISRALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSL QPEDFATYYCQQFNSYPITFGQGTRLEIKRT SEQ ID NO: 5 (nucleotides encoding an amino acid sequence of SEQ ID NO: 9) CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTC CCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGCCA TGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGACATTT ATATGGGATGATGGAAGTTTTAAATATTATGCAGAGTCCGTGAAGGGCCG ATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATTTGCAAATGA ACAGCCTGAGAGCCGAAGACACGGCTGTGTATTACTGTGCGAGAGAGGGC AGCTGGTCTCCTGATATATTTGATATCTGGGGCCAAGGGACAATGGTCAC CGTCTCTTCA SEQ ID NO: 6 (nucleotides encoding an amino acid sequence of SEQ ID NO: 2) GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGA CAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGAGCTTTAG CCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATC TGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTG CAACTTATTACTGTCAACAGTTTAATAGTTACCCCATCACCTTCGGCCAA GGGACACGACTGGAGATTAAACGAACT SEQ ID NO: 7 (nucleotides encoding an amino acid sequence of SEQ ID NO: 3) ATGGAGTTTGGGCTGAACTGGGTTTTCCTCGTTGCTCTTTTCAGAGGTGT CCACTGTCAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTG GGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGC TATGCCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGT GACATTTATATGGGATGATGGAAGTTTTAAATATTATGCAGAGTCCGTGA AGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATTTG CAAATGAACAGCCTGAGAGCCGAAGACACGGCTGTGTATTACTGTGCGAG AGAGGGCAGCTGGTCTCCTGATATATTTGATATCTGGGGCCAAGGGACAA TGGTCACCGTCTCTTCA SEQ ID NO: 8 (nucleotides encoding an amino acid sequence of SEQ ID NO: 4) ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCT CCCAGGTGCCAGATGTGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT CTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC ATTAGCAGAGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAA GCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGT TCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTG CAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCC CATCACCTTCGGCCAAGGGACACGACTGGAGATTAAACGAACT SEQ ID NO: 9 (amino acid sequence of the mature polypeptide of the heavy chain, underlined amino acids correspond to CDR1, CDR2 and CDR3) QVQLVESGGGVVQPGRSLRLSCAASGFTLSSFGMHWVRQAPGKGLEWVAV IWYDGSNEYYADSVKGRFTTSRDNSKNTLYLQMNSLRAEDTAVYYCAREG SWSPDIFDIWGQGTMVTVSS SEQ ID NO: 10 (amino acid sequence of the mature polypeptide of the light chain, underlined amino acids correspond to CDR1, CDR2' and CDR3) AIQLTQSPSSLSASVGDRVTITCRTSQGIRSALAWYQQNPGKAPKLLIYD ASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPVTFGQ GTRLEIKRT SEQ ID NO: 11 (amino acid sequence of the polypeptide of SEQ ID NO: 9 with leader sequence, amino acids marked in bold correspond to the leader sequence) MEFGLSWVFLVALLRGVQCQVQLVESGGGVVQPGRSLRLSCAASGFTLSS FGMHWVRQAPGKGLEWVAVIWYDGSNEYYADSVKGRFTTSRDNSKNTLYL QMNSLRAEDTAVYYCAREGSWSPDIFDIWGQGTMVTVSS SEQ ID NO: 12 (amino acid sequence of the polypeptide of SEQ ID NO: 10 with leader sequence, amino acids marked in bold correspond to the leader sequence) MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRTSQG IRSALAWYQQNPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSL QPEDFATYYCQQFNSYPVTFGQGTRLEIKRT SEQ ID NO: 13 (nucleotides encoding an amino acid sequence of SEQ ID NO: 9) CAGGTGCAACTGGTGGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTC CCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTAAGTAGCTTTGGCA TGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTT ATATGGTATGATGGAAGTAATGAATACTATGCAGACTCCGTGAAGGGCCG ATTCACCACCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGA ACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAGGGC AGCTGGTCTCCTGATATTTTTGATATCTGGGGCCAAGGGACAATGGTCAC CGTCTCTTCA SEQ ID NO: 14 (nucleotides encoding an amino acid sequence of SEQ ID NO: 10) GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGA CAGAGTCACCATCACTTGCCGGACAAGTCAGGGCATTCGCAGTGCTTTAG CCTGGTATCAGCAGAACCCCGGGAAAGCTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATC TGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTG CAACTTATTACTGTCAACAGTTTAATAGTTACCCCGTCACCTTCGGCCAA GGGACACGACTGGAGATTAAACGAACT SEQ ID NO: 15 (nucleotides encoding an amino acid sequence of SEQ ID NO: 11) ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTAAGAGGTGT CCAGTGTCAGGTGCAACTGGTGGAGTCGGGGGGAGGCGTGGTCCAGCCTG GGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTAAGTAGC TTTGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGT GGCAGTTATATGGTATGATGGAAGTAATGAATACTATGCAGACTCCGTGA AGGGCCGATTCACCACCTCCAGAGACAATTCCAAGAACACGCTGTATCTG CAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAG AGAGGGCAGCTGGTCTCCTGATATTTTTGATATCTGGGGCCAAGGGACAA TGGTCACCGTCTCTTCA SEQ ID NO: 16 (nucleotides encoding an amino acid sequence of SEQ ID NO: 12) ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCT CCCAGGTGCCAGATGTGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT CTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGACAAGTCAGGGC ATTCGCAGTGCTTTAGCCTGGTATCAGCAGAACCCCGGGAAAGCTCCTAA GCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGT TCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTG CAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCC CGTCACCTTCGGCCAAGGGACACGACTGGAGATTAAACGAACT SEQ ID NO: 17 (amino acid sequence of the mature polypeptide of the heavy chain, underlined amino acids correspond to CDR1, CDR2 and CDR3) QVQLVESGGGMVQPGRSLRLSCAASGFTLSSYGMHWVRQAPGKGLEWVTV IWYDGNNQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREG SWSPDIFDIWGQGTMVTVSS SEQ ID NO: 18 (amino acid sequence of the mature polypeptide of the light chain, underlined amino acids correspond to CDR1, CDR2' and CDR3) AIQLTQSPSSLSASVGDRVTITCRASQGISSYLAWYQQKPGKAPKLLIYD ASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPITFGQ

GTRLEIKRT SEQ ID NO: 19 (amino acid sequence of the polypeptide of SEQ ID NO: 17 with leader sequence, amino acids marked in bold correspond to the leader sequence) MEFGLSWVFLVALLRGVQCQVQLVESGGGMVQPGRSLRLSCAASGFTLSS YGMHWVRQAPGKGLEWVTVIWYDGNNQYYADSVKGRFTISRDNSKNTLYL QMNSLRAEDTAVYYCAREGSWSPDIFDIWGQGTMVTVSS SEQ ID NO: 20 (amino acid sequence of the polypeptide of SEQ ID NO: 18 with leader sequence, amino acids marked in bold correspond to the leader sequence) MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRASQG ISSYLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSL QPEDFATYYCQQFNSYPITFGQGTRLEIKRT SEQ ID NO: 21 (nucleotides encoding an amino acid sequence of SEQ ID NO: 17) CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCATGGTCCAGCCTGGGAGGTC CCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCCTCAGTAGCTATGGCA TGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGACAGTT ATATGGTATGATGGAAATAATCAATACTATGCAGACTCCGTGAAGGGCCG ATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGA ACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAGAGAGGGC AGCTGGTCTCCTGATATTTTTGATATCTGGGGCCAAGGGACAATGGTCAC CGTCTCTTCA SEQ ID NO: 22 (nucleotides encoding an amino acid sequence of SEQ ID NO: 18) GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGA CAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTTATTTAG CCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATC TGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTG CAACTTATTACTGTCAACAGTTTAATAGTTACCCCATCACCTTCGGCCAA GGGACACGACTGGAGATTAAACGAACT SEQ ID NO: 23 (nucleotides encoding an amino acid sequence of SEQ ID NO: 19) ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTAAGAGGTGT CCAGTGTCAGGTGCAGCTGGTGGAGTCTGGGGGAGGCATGGTCCAGCCTG GGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCCTCAGTAGC TATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGT GACAGTTATATGGTATGATGGAAATAATCAATACTATGCAGACTCCGTGA AGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTG CAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAG AGAGGGCAGCTGGTCTCCTGATATTTTTGATATCTGGGGCCAAGGGACAA TGGTCACCGTCTCTTCA SEQ ID NO: 24 (nucleotides encoding an amino acid sequence of SEQ ID NO: 20) ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCT CCCAGGTGCCAGATGTGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT CTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC ATTAGCAGTTATTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAA GCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGT TCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTG CAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCC CATCACCTTCGGCCAAGGGACACGACTGGAGATTAAACGAACT SEQ ID NO: 25 (amino acid sequence of the mature polypeptide of the heavy chain, underlined amino acids correspond to CDR1, CDR2 and CDR3) QVQLVQSGAEVKKPGSSVKVSCKASGDTFSSYAISWVRQAPGQGFEWMGG IIPVIGTVNYEERFQDRVTITADNSTSTAYMELTSLRSEDTAVYFCGREE GFLDYWGQGTLVTVSS SEQ ID NO: 26 (amino acid sequence of the mature polypeptide of the light chain, underlined amino acids correspond to CDR 1, CDR2' and CDR3) AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYD ASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPLLTFG GGTKVEIKRT SEQ ID NO: 27 (amino acid sequence of the polypeptide of SEQ ID NO: 25 with leader sequence, amino acids marked in bold correspond to the leader sequence) MDWTWRFLFVVAAATGVQSQVQLVQSGAEVKKPGSSVKVSCKASGDTFSS YAISWVRQAPGQGFEWMGGIIPVIGTVNYEERFQDRVTITADNSTSTAYM ELTSLRSEDTAVYFCGREEGFLDYWGQGTLVTVSS SEQ ID NO: 28 (amino acid sequence of the polypeptide of SEQ ID NO: 26 with leader sequence, amino acids marked in bold correspond to the leader sequence) MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRASQG ISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSL QPEDFATYYCQQFNSYPLLTFGGGTKVEIKRT SEQ ID NO: 29 (nucleotides encoding an amino acid sequence of SEQ ID NO: 25) CAGGTCCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTC GGTGAAGGTCTCCTGCAAGGCTTCTGGAGACACCTTCAGCAGTTATGCTA TCAGTTGGGTGCGACAGGCCCCTGGACAAGGGTTTGAGTGGATGGGAGGG ATCATCCCTGTCATTGGTACAGTAAATTATGAAGAGAGATTCCAGGACAG AGTCACGATTACCGCGGACAATTCCACGAGCACAGCCTACATGGAGTTGA CTAGTCTGAGATCTGAAGACACGGCCGTGTATTTTTGTGGGAGAGAAGAG GGCTTCCTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA SEQ ID NO: 30 (nucleotides encoding an amino acid sequence of SEQ ID NO: 26) ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCT CCCAGGTGCCAGATGTGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT CTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC ATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAA GCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGT TCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTG CAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCC TCTTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACG SEQ ID NO: 31 (nucleotides encoding an amino acid sequence of SEQ ID NO: 27) ATGGACTGGACCTGGAGGTTCCTCTTTGTGGTGGCAGCAGCTACAGGTGT CCAGTCCCAGGTCCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTG GGTCCTCGGTGAAGGTCTCCTGCAAGGCTTCTGGAGACACCTTCAGCAGT TATGCTATCAGTTGGGTGCGACAGGCCCCTGGACAAGGGTTTGAGTGGAT GGGAGGGATCATCCCTGTCATTGGTACAGTAAATTATGAAGAGAGATTCC AGGACAGAGTCACGATTACCGCGGACAATTCCACGAGCACAGCCTACATG GAGTTGACTAGTCTGAGATCTGAAGACACGGCCGTGTATTTTTGTGGGAG AGAAGAGGGCTTCCTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCT CCTCA SEQ ID NO: 32 (nucleotides encoding an amino acid sequence of SEQ ID NO: 28) ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCT CCCAGGTGCCAGATGTGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT CTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC ATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAA GCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGT TCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTG CAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCC TCTTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACG SEQ ID NO: 33 (amino acid sequence of the mature polypeptide of the heavy chain, underlined amino acids correspond to CDR1, CDR2 and CDR3) QVQLVESGGGVVQPGRSLRLSCAASGFTFSCCGMHWVRQAPGKGLEWVAV IWYDGSNKYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCATDS SGSFYEYFQHWGQGTLVTVSS SEQ ID NO: 34 (amino acid sequence of the mature polypeptide of the light chain, underlined amino acids correspond to CDR1, CDR2' and CDR3) AIQLTQSPSSLSASVGDRVTITCRASQGINSALAWYQQKPGKAPKLLIYD ASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPYTFGQ GTKLEIKRT SEQ ID NO: 35 (amino acid sequence of the polypeptide of SEQ ID NO: 33 with leader sequence, amino acids marked in bold correspond to the leader sequence) MEFGLSWVFLVALLRGVQCQVQLVESGGGVVQPGRSLRLSCAASGFTFSC CGMHWVRQAPGKGLEWVAVIWYDGSNKYYADSVKGRFTISRDTSKNTLYL QMNSLRAEDTAVYYCATDSSGSFYEYFQHWGQGTLVTVSS SEQ ID NO: 36 (amino acid sequence of the polypeptide of SEQ ID NO: 34 with leader sequence, amino acids marked in bold correspond to the leader sequence) MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRASQG INSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSL

QPEDFATYYCQQFNSYPYTFGQGTKLEIKRT SEQ ID NO: 37 (nucleotides encoding an amino acid sequence of SEQ ID NO: 33) CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTC CCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTTGCTGTGGCA TGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTT ATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCG ATTCACCATCTCCAGAGACACTTCCAAGAACACGCTGTATCTGCAAATGA ACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGACAGATAGT TCGGGGAGTTTTTATGAATACTTCCAGCACTGGGGCCAGGGCACCCTGGT CACCGTCTCCTCA SEQ ID NO: 38 (nucleotides encoding an amino acid sequence of SEQ ID NO: 34) GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGA CAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAACAGTGCTTTAG CCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATC TGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTG CAACTTATTACTGTCAACAGTTTAATAGTTACCCGTACACTTTTGGCCAG GGGACCAAGCTGGAGATCAAACGAACT SEQ ID NO: 39 (nucleotides encoding an amino acid sequence of SEQ ID NO: 35) ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTAAGAGGTGT CCAGTGTCAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTG GGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTTGC TGTGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGT GGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGA AGGGCCGATTCACCATCTCCAGAGACACTTCCAAGAACACGCTGTATCTG CAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAC AGATAGTTCGGGGAGTTTTTATGAATACTTCCAGCACTGGGGCCAGGGCA CCCTGGTCACCGTCTCCTCA SEQ ID NO: 40 (nucleotides encoding an amino acid sequence of SEQ ID NO: 36) ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCT CCCAGGTGCCAGATGTGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT CTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC ATTAACAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAA GCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGT TCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTG CAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCC GTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAACGAACT SEQ ID NO: 41 (amino acid sequence of the mature polypeptide of the heavy chain, underlined amino acids correspond to CDR1, CDR2 and CDR3) QVQLVESGGGVVQPGRSLRLSCAASGFTFSGYGMHWVRQAPGRGLDWVAV VWYDGGYKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDS SGSFYEYLQHWGQGTLVTVSS SEQ ID NO: 42 (amino acid sequence of the mature polypeptide of the light chain, underlined amino acids correspond to CDR1, CDR2' and CDR3) AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPHFW PGDQAGDQTNCG SEQ ID NO: 43 (amino acid sequence of the polypeptide of SEQ ID NO: 41 with leader sequence, amino acids marked in bold correspond to the leader sequence) MEFGLSWVFLVALLRGVQCQVQLVESGGGVVQPGRSLRLSCAASGFTFSG YGMHWVRQAPGRGLDWVAVVWYDGGYKFYADSVKGRFTISRDNSKNTLYL QMNSLRAEDTAVYYCARDSSGSFYEYLQHWGQGTLVTVSS SEQ ID NO: 44 (amino acid sequence of the polypeptide of SEQ ID NO: 42 with leader sequence, amino acids marked in bold correspond to the leader sequence) MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRASQG ISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSL QPEDFATYYCQQFNSYPHFWPGDQAGDQTNCG SEQ ID NO: 45 (nucleotides encoding an amino acid sequence of SEQ ID NO: 41) CAGGTGCAGTTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTC CCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTGGCTATGGCA TGCACTGGGTCCGCCAGGCTCCAGGCAGGGGGCTGGACTGGGTGGCAGTT GTGTGGTATGATGGAGGTTATAAGTTCTATGCAGACTCCGTGAAGGGCCG ATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGA ACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGATAGT TCGGGGAGTTTTTATGAATACTTACAACATTGGGGCCAGGGCACCCTGGT CACCGTCTCCTCA SEQ ID NO: 46 (nucleotides encoding an amino acid sequence of SEQ ID NO: 42) GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGA CAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAG CCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATC TGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTG CAACTTATTACTGTCAACAGTTTAATAGTTACCCTCACTTTTGGCCAGGG GACCAAGCTGGAGATCAAACGAACT SEQ ID NO: 47 (nucleotides encoding an amino acid sequence of SEQ ID NO: 43) ATGGAGTTTGGGCTGAGCTGGGTTTTCCTCGTTGCTCTTTTAAGAGGTGT CCAGTGTCAGGTGCAGTTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTG GGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTGGC TATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAGGGGGCTGGACTGGGT GGCAGTTGTGTGGTATGATGGAGGTTATAAGTTCTATGCAGACTCCGTGA AGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTG CAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAG AGATAGTTCGGGGAGTTTTTATGAATACTTACAACATTGGGGCCAGGGCA CCCTGGTCACCGTCTCCTCA SEQ ID NO: 48 (nucleotides encoding an amino acid sequence of SEQ ID NO: 44) ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCT CCCAGGTGCCAGATGTGCCATCCAGTTGACCCAGTCTCCATCCTCCCTGT CTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC ATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAA GCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGT TCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTG CAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCC TCACTTTTGGCCAGGGGACCAAGCTGGAGATCAAACGAACT

Sequence CWU 1

1

791120PRTHomo Sapiens 1Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Phe Ile Trp Asp Asp Gly Ser Phe Lys Tyr Tyr Ala Glu Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Gly Ser Trp Ser Pro Asp Ile Phe Asp Ile Trp Gly Gln 100 105 110Gly Thr Met Val Thr Val Ser Ser 115 1202109PRTHomo Sapiens 2Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Arg Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Ile 85 90 95Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr 100 1053139PRTHomo Sapiens 3Met Glu Phe Gly Leu Asn Trp Val Phe Leu Val Ala Leu Phe Arg Gly1 5 10 15Val His Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln 20 25 30Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45Ser Ser Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60Glu Trp Val Thr Phe Ile Trp Asp Asp Gly Ser Phe Lys Tyr Tyr Ala65 70 75 80Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 85 90 95Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Ala Arg Glu Gly Ser Trp Ser Pro Asp Ile Phe Asp Ile 115 120 125Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 130 1354131PRTHomo Sapiens 4Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp1 5 10 15Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45Gln Gly Ile Ser Arg Ala Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val65 70 75 80Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110Phe Asn Ser Tyr Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile 115 120 125Lys Arg Thr 1305360DNAHomo Sapiens 5caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt agctatgcca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtgacattt atatgggatg atggaagttt taaatattat 180gcagagtccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ttgcaaatga acagcctgag agccgaagac acggctgtgt attactgtgc gagagagggc 300agctggtctc ctgatatatt tgatatctgg ggccaaggga caatggtcac cgtctcttca 3606327DNAHomo Sapiens 6gccatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattagc agagctttag cctggtatca gcagaaacca 120gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag tttaatagtt accccatcac cttcggccaa 300gggacacgac tggagattaa acgaact 3277417DNAHomo Sapiens 7atggagtttg ggctgaactg ggttttcctc gttgctcttt tcagaggtgt ccactgtcag 60gtgcagctgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gagactctcc 120tgtgcagcgt ctggattcac cttcagtagc tatgccatgc actgggtccg ccaggctcca 180ggcaaggggc tggagtgggt gacatttata tgggatgatg gaagttttaa atattatgca 240gagtccgtga agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatttg 300caaatgaaca gcctgagagc cgaagacacg gctgtgtatt actgtgcgag agagggcagc 360tggtctcctg atatatttga tatctggggc caagggacaa tggtcaccgt ctcttca 4178393DNAHomo Sapiens 8atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccgggc aagtcagggc attagcagag ctttagcctg gtatcagcag 180aaaccaggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc catcaccttc 360ggccaaggga cacgactgga gattaaacga act 3939120PRTHomo Sapiens 9Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Ser Phe 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Thr Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Gly Ser Trp Ser Pro Asp Ile Phe Asp Ile Trp Gly Gln 100 105 110Gly Thr Met Val Thr Val Ser Ser 115 12010109PRTHomo Sapiens 10Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Gly Ile Arg Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln Asn Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Val 85 90 95Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr 100 10511139PRTHomo Sapiens 11Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Leu Leu Arg Gly1 5 10 15Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln 20 25 30Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu 35 40 45Ser Ser Phe Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala65 70 75 80Asp Ser Val Lys Gly Arg Phe Thr Thr Ser Arg Asp Asn Ser Lys Asn 85 90 95Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Ala Arg Glu Gly Ser Trp Ser Pro Asp Ile Phe Asp Ile 115 120 125Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 130 13512131PRTHomo Sapiens 12Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp1 5 10 15Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Thr Ser 35 40 45Gln Gly Ile Arg Ser Ala Leu Ala Trp Tyr Gln Gln Asn Pro Gly Lys 50 55 60Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val65 70 75 80Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110Phe Asn Ser Tyr Pro Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile 115 120 125Lys Arg Thr 13013360DNAHomo Sapiens 13caggtgcaac tggtggagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttaagt agctttggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatggtatg atggaagtaa tgaatactat 180gcagactccg tgaagggccg attcaccacc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagagagggc 300agctggtctc ctgatatttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 36014327DNAHomo Sapiens 14gccatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc ggacaagtca gggcattcgc agtgctttag cctggtatca gcagaacccc 120gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag tttaatagtt accccgtcac cttcggccaa 300gggacacgac tggagattaa acgaact 32715417DNAHomo Sapiens 15atggagtttg ggctgagctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag 60gtgcaactgg tggagtcggg gggaggcgtg gtccagcctg ggaggtccct gagactctcc 120tgtgcagcgt ctggattcac cttaagtagc tttggcatgc actgggtccg ccaggctcca 180ggcaaggggc tggagtgggt ggcagttata tggtatgatg gaagtaatga atactatgca 240gactccgtga agggccgatt caccacctcc agagacaatt ccaagaacac gctgtatctg 300caaatgaaca gcctgagagc cgaggacacg gctgtgtatt actgtgcgag agagggcagc 360tggtctcctg atatttttga tatctggggc caagggacaa tggtcaccgt ctcttca 41716393DNAHomo Sapiens 16atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccggac aagtcagggc attcgcagtg ctttagcctg gtatcagcag 180aaccccggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc cgtcaccttc 360ggccaaggga cacgactgga gattaaacga act 39317120PRTHomo Sapiens 17Gln Val Gln Leu Val Glu Ser Gly Gly Gly Met Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly Asn Asn Gln Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Gly Ser Trp Ser Pro Asp Ile Phe Asp Ile Trp Gly Gln 100 105 110Gly Thr Met Val Thr Val Ser Ser 115 12018109PRTHomo Sapiens 18Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Ile 85 90 95Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr 100 10519139PRTHomo Sapiens 19Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Leu Leu Arg Gly1 5 10 15Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Met Val Gln 20 25 30Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu 35 40 45Ser Ser Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60Glu Trp Val Thr Val Ile Trp Tyr Asp Gly Asn Asn Gln Tyr Tyr Ala65 70 75 80Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 85 90 95Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Ala Arg Glu Gly Ser Trp Ser Pro Asp Ile Phe Asp Ile 115 120 125Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 130 13520131PRTHomo Sapiens 20Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp1 5 10 15Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45Gln Gly Ile Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val65 70 75 80Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110Phe Asn Ser Tyr Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile 115 120 125Lys Arg Thr 13021360DNAHomo Sapiens 21caggtgcagc tggtggagtc tgggggaggc atggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccctcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtgacagtt atatggtatg atggaaataa tcaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtat attactgtgc gagagagggc 300agctggtctc ctgatatttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 36022327DNAHomo Sapiens 22gccatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattagc agttatttag cctggtatca gcagaaacca 120gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag tttaatagtt accccatcac cttcggccaa 300gggacacgac tggagattaa acgaact 32723417DNAHomo Sapiens 23atggagtttg ggctgagctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag 60gtgcagctgg tggagtctgg gggaggcatg gtccagcctg ggaggtccct gagactctcc 120tgtgcagcgt ctggattcac cctcagtagc tatggcatgc actgggtccg ccaggctcca 180ggcaaggggc tggagtgggt gacagttata tggtatgatg gaaataatca atactatgca 240gactccgtga agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctg 300caaatgaaca gcctgagagc cgaggacacg gctgtatatt actgtgcgag agagggcagc 360tggtctcctg atatttttga tatctggggc caagggacaa tggtcaccgt ctcttca 41724393DNAHomo Sapiens 24atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccgggc aagtcagggc attagcagtt atttagcctg gtatcagcag 180aaaccaggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc catcaccttc 360ggccaaggga cacgactgga gattaaacga act 39325116PRTHomo Sapiens 25Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Asp Thr Phe Ser Ser Tyr 20 25 30Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Phe Glu Trp Met 35 40 45Gly Gly Ile Ile Pro Val Ile Gly Thr Val Asn Tyr Glu Glu Arg Phe 50 55 60Gln Asp Arg Val Thr Ile Thr Ala Asp Asn Ser Thr Ser Thr Ala Tyr65 70 75 80Met Glu Leu Thr Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95Gly Arg Glu Glu Gly Phe Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 11526110PRTHomo Sapiens 26Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala

20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu 85 90 95Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 11027135PRTHomo Sapiens 27Met Asp Trp Thr Trp Arg Phe Leu Phe Val Val Ala Ala Ala Thr Gly1 5 10 15Val Gln Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys 20 25 30Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Asp Thr Phe 35 40 45Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Phe 50 55 60Glu Trp Met Gly Gly Ile Ile Pro Val Ile Gly Thr Val Asn Tyr Glu65 70 75 80Glu Arg Phe Gln Asp Arg Val Thr Ile Thr Ala Asp Asn Ser Thr Ser 85 90 95Thr Ala Tyr Met Glu Leu Thr Ser Leu Arg Ser Glu Asp Thr Ala Val 100 105 110Tyr Phe Cys Gly Arg Glu Glu Gly Phe Leu Asp Tyr Trp Gly Gln Gly 115 120 125Thr Leu Val Thr Val Ser Ser 130 13528132PRTHomo Sapiens 28Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp1 5 10 15Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45Gln Gly Ile Ser Ser Ala Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val65 70 75 80Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110Phe Asn Ser Tyr Pro Leu Leu Thr Phe Gly Gly Gly Thr Lys Val Glu 115 120 125Ile Lys Arg Thr 13029348DNAHomo Sapiens 29caggtccagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg cttctggaga caccttcagc agttatgcta tcagttgggt gcgacaggcc 120cctggacaag ggtttgagtg gatgggaggg atcatccctg tcattggtac agtaaattat 180gaagagagat tccaggacag agtcacgatt accgcggaca attccacgag cacagcctac 240atggagttga ctagtctgag atctgaagac acggccgtgt atttttgtgg gagagaagag 300ggcttccttg actattgggg ccagggaacc ctggtcaccg tctcctca 34830396DNAHomo Sapiens 30atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccgggc aagtcagggc attagcagtg ctttagcctg gtatcagcag 180aaaccaggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc tcttctcact 360ttcggcggag ggaccaaggt ggagatcaaa cgtacg 39631405DNAHomo Sapiens 31atggactgga cctggaggtt cctctttgtg gtggcagcag ctacaggtgt ccagtcccag 60gtccagctgg tgcagtctgg ggctgaggtg aagaagcctg ggtcctcggt gaaggtctcc 120tgcaaggctt ctggagacac cttcagcagt tatgctatca gttgggtgcg acaggcccct 180ggacaagggt ttgagtggat gggagggatc atccctgtca ttggtacagt aaattatgaa 240gagagattcc aggacagagt cacgattacc gcggacaatt ccacgagcac agcctacatg 300gagttgacta gtctgagatc tgaagacacg gccgtgtatt tttgtgggag agaagagggc 360ttccttgact attggggcca gggaaccctg gtcaccgtct cctca 40532396DNAHomo Sapiens 32atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccgggc aagtcagggc attagcagtg ctttagcctg gtatcagcag 180aaaccaggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc tcttctcact 360ttcggcggag ggaccaaggt ggagatcaaa cgtacg 39633121PRTHomo Sapiens 33Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Cys Cys 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Thr Asp Ser Ser Gly Ser Phe Tyr Glu Tyr Phe Gln His Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val Ser Ser 115 12034109PRTHomo Sapiens 34Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asn Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100 10535140PRTHomo Sapiens 35Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Leu Leu Arg Gly1 5 10 15Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln 20 25 30Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45Ser Cys Cys Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala65 70 75 80Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser Lys Asn 85 90 95Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Ala Thr Asp Ser Ser Gly Ser Phe Tyr Glu Tyr Phe Gln 115 120 125His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 130 135 14036131PRTHomo Sapiens 36Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp1 5 10 15Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45Gln Gly Ile Asn Ser Ala Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val65 70 75 80Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110Phe Asn Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 115 120 125Lys Arg Thr 13037363DNAHomo Sapiens 37caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt tgctgtggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatggtatg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca cttccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gacagatagt 300tcggggagtt tttatgaata cttccagcac tggggccagg gcaccctggt caccgtctcc 360tca 36338327DNAHomo Sapiens 38gccatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaac agtgctttag cctggtatca gcagaaacca 120gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag tttaatagtt acccgtacac ttttggccag 300gggaccaagc tggagatcaa acgaact 32739420DNAHomo Sapiens 39atggagtttg ggctgagctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag 60gtgcagctgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gagactctcc 120tgtgcagcgt ctggattcac cttcagttgc tgtggcatgc actgggtccg ccaggctcca 180ggcaaggggc tggagtgggt ggcagttata tggtatgatg gaagtaataa atactatgca 240gactccgtga agggccgatt caccatctcc agagacactt ccaagaacac gctgtatctg 300caaatgaaca gcctgagagc cgaggacacg gctgtgtatt actgtgcgac agatagttcg 360gggagttttt atgaatactt ccagcactgg ggccagggca ccctggtcac cgtctcctca 42040393DNAHomo Sapiens 40atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccgggc aagtcagggc attaacagtg ctttagcctg gtatcagcag 180aaaccaggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc gtacactttt 360ggccagggga ccaagctgga gatcaaacga act 39341121PRTHomo Sapiens 41Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Asp Trp Val 35 40 45Ala Val Val Trp Tyr Asp Gly Gly Tyr Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Ser Ser Gly Ser Phe Tyr Glu Tyr Leu Gln His Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val Ser Ser 115 12042110PRTHomo Sapiens 42Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro His 85 90 95Phe Trp Pro Gly Asp Gln Ala Gly Asp Gln Thr Asn Cys Gly 100 105 11043140PRTHomo Sapiens 43Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Leu Leu Arg Gly1 5 10 15Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln 20 25 30Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45Ser Gly Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Arg Gly Leu 50 55 60Asp Trp Val Ala Val Val Trp Tyr Asp Gly Gly Tyr Lys Phe Tyr Ala65 70 75 80Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 85 90 95Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110Tyr Tyr Cys Ala Arg Asp Ser Ser Gly Ser Phe Tyr Glu Tyr Leu Gln 115 120 125His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 130 135 14044132PRTHomo Sapiens 44Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp1 5 10 15Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45Gln Gly Ile Ser Ser Ala Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val65 70 75 80Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110Phe Asn Ser Tyr Pro His Phe Trp Pro Gly Asp Gln Ala Gly Asp Gln 115 120 125Thr Asn Cys Gly 13045363DNAHomo Sapiens 45caggtgcagt tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt ggctatggca tgcactgggt ccgccaggct 120ccaggcaggg ggctggactg ggtggcagtt gtgtggtatg atggaggtta taagttctat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagagatagt 300tcggggagtt tttatgaata cttacaacat tggggccagg gcaccctggt caccgtctcc 360tca 36346325DNAHomo Sapiens 46gccatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca 120gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag tttaatagtt accctcactt ttggccaggg 300gaccaagctg gagatcaaac gaact 32547420DNAHomo Sapiens 47atggagtttg ggctgagctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag 60gtgcagttgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gagactctcc 120tgtgcagcgt ctggattcac cttcagtggc tatggcatgc actgggtccg ccaggctcca 180ggcagggggc tggactgggt ggcagttgtg tggtatgatg gaggttataa gttctatgca 240gactccgtga agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctg 300caaatgaaca gcctgagagc cgaggacacg gctgtgtatt actgtgcgag agatagttcg 360gggagttttt atgaatactt acaacattgg ggccagggca ccctggtcac cgtctcctca 42048391DNAHomo Sapiens 48atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccgggc aagtcagggc attagcagtg ctttagcctg gtatcagcag 180aaaccaggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc tcacttttgg 360ccaggggacc aagctggaga tcaaacgaac t 3914910PRTHomo Sapiens 49Gly Phe Thr Phe Ser Ser Tyr Ala Met His1 5 105017PRTHomo Sapiens 50Phe Ile Trp Asp Asp Gly Ser Phe Lys Tyr Tyr Ala Glu Ser Val Lys1 5 10 15Gly519PRTHomo Sapiens 51Glu Gly Ser Trp Ser Pro Asp Ile Phe1 5527PRTHomo Sapiens 52Ser Gln Gly Ile Ser Arg Ala1 5533PRTHomo Sapiens 53Asp Ala Ser1546PRTHomo Sapiens 54Phe Asn Ser Tyr Pro Ile1 55510PRTHomo Sapiens 55Gly Phe Thr Leu Ser Ser Phe Gly Met His1 5 105617PRTHomo Sapiens 56Val Ile Trp Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly579PRTHomo Sapiens 57Glu Gly Ser Trp Ser Pro Asp Ile Phe1 5587PRTHomo Sapiens 58Ser Gln Gly Ile Arg Ser Ala1 5596PRTHomo Sapiens 59Phe Asn Ser Tyr Pro Val1 56010PRTHomo Sapiens 60Gly Phe Thr Leu Ser Ser Tyr Gly Met His1 5 106117PRTHomo Sapiens 61Val Ile Trp Tyr Asp Gly Asn Asn Gln Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly629PRTHomo Sapiens 62Glu Gly Ser Trp Ser Pro Asp Ile Phe1 5637PRTHomo Sapiens 63Ser Gln Gly Ile Ser Ser Tyr1 5645PRTHomo Sapiens 64Phe Asn Ser Tyr Pro1 56510PRTHomo Sapiens 65Gly Asp Thr Phe Ser Ser Tyr Ala Ile Ser1 5 106617PRTHomo Sapiens 66Gly Ile Ile Pro Val Ile Gly Thr Val Asn Tyr Glu Glu Arg Phe Gln1 5 10 15Asp675PRTHomo Sapiens 67Glu Glu Gly Phe Leu1 5687PRTHomo Sapiens 68Ser Gln Gly Ile Ser Ser Ala1 5696PRTHomo Sapiens 69Phe Asn Ser Tyr Pro Leu1 57010PRTHomo Sapiens 70Gly Phe Thr Phe Ser Cys Cys Gly Met His1 5

107117PRTHomo Sapiens 71Val Ile Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly7210PRTHomo Sapiens 72Asp Ser Ser Gly Ser Phe Tyr Glu Tyr Phe1 5 10737PRTHomo Sapiens 73Ser Gln Gly Ile Asn Ser Ala1 5746PRTHomo Sapiens 74Phe Asn Ser Tyr Pro Tyr1 57510PRTHomo Sapiens 75Gly Phe Thr Phe Ser Gly Tyr Gly Met His1 5 107617PRTHomo Sapiens 76Val Val Trp Tyr Asp Gly Gly Tyr Lys Phe Tyr Ala Asp Ser Val Lys1 5 10 15Gly7710PRTHomo Sapiens 77Asp Ser Ser Gly Ser Phe Tyr Glu Tyr Leu1 5 10787PRTHomo Sapiens 78Ser Gln Gly Ile Ser Ser Ala1 5796PRTHomo Sapiens 79Phe Asn Ser Tyr Pro His1 5

Claims

1. An isolated human IL-4 specific antibody, or fragment thereof, which binds to human IL-4 with a dissociation constant K.sub.d of equal or less than 800 pM.

2. An antibody of claim 1 comprising a) a polypeptide of SEQ ID NO:1 and a polypeptide of SEQ ID NO:2, or b) a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:10, or c) a polypeptide of SEQ ID NO:17 and a polypeptide of SEQ ID NO:18, or d) a polypeptide of SEQ ID NO:25 and a polypeptide of SEQ ID NO:26, or e) a polypeptide of SEQ ID NO:33 and a polypeptide of SEQ ID NO:34, or f) a polypeptide of SEQ ID NO:41 and a polypeptide of SEQ ID NO:42.

3. An antibody of claim 1 comprising a) a polypeptide of SEQ ID NO:3 and a polypeptide of SEQ ID NO:4, or b) a polypeptide of SEQ ID NO:11 and a polypeptide of SEQ ID NO:12, or c) a polypeptide of SEQ ID NO:19 and a polypeptide of SEQ ID NO:20, or d) a polypeptide of SEQ ID NO:27 and a polypeptide of SEQ ID NO:28, or e) a polypeptide of SEQ ID NO:35 and a polypeptide of SEQ ID NO:36, or f) a polypeptide of SEQ ID NO:43 and a polypeptide of SEQ ID NO:44.

4. The antibody of claim 1 that is monoclonal.

5. Isolated polynucleotides comprising polynucleotides encoding an antibody of claim 1.

6. Polynucleotides of claim 6 encoding the amino acid sequence of an antibody of claim 2.

7. Polynucleotides of claim 6 comprising a) a polynucleotide of SEQ ID NO:5 and a polynucleotide of SEQ ID NO:6, or b) a polynucleotide of SEQ ID NO:13 and a polynucleotide of SEQ ID NO:14, or c) a polynucleotide of SEQ ID NO:21 and a polynucleotide of SEQ ID NO:22, or d) a polynucleotide of SEQ ID NO:29 and a polynucleotide of SEQ ID NO:30, or e) a polynucleotide of SEQ ID NO:37 and a polynucleotide of SEQ ID NO:38, or f) a polynucleotide of SEQ ID NO:45 and a polynucleotide of SEQ ID NO:46.

8. Polynucleotides of claim 6 encoding a) a polypeptide of SEQ ID NO:7 and SEQ ID NO:8, or b) a polypeptide of SEQ ID NO:15 and SEQ ID NO:16, or c) a polypeptide of SEQ ID NO:23 and SEQ ID NO:24, or d) a polypeptide of SEQ ID NO:31 and SEQ ID NO:32, or e) a polypeptide of SEQ ID NO:39 and SEQ ID NO:40, or f) a polypeptide of SEQ ID NO:47 and SEQ ID NO:48.

9. An expression vector comprising polynucleotides of claim 6.

10. An expression system comprising a polynucleotide of claims 6 wherein said expression system or part thereof is capable of producing an antibody of claim 1 when said expression system or part thereof is present in a compatible host cell.

11. An isolated host cell comprising an expression system of claim 11.

12. Use of an antibody of any one of claims 1 to 5 as a pharmaceutical.

13. Use of an antibody of any one of claims 1 to 5 for the manufacture of a medicament for the treatment of diseases mediated by IL-4 and/or IgE.

14. Use of claim 14 wherein the disease is selected from the group consisting of atopic dermatitis, allergic asthma and allergic rhinitis.

15. Use of any one of claims 13 to 15 wherein the antibody is selected from the group consisting of an human IL-4 monoclonal antibody, a fragment thereof and an analog thereof.

16. A pharmaceutical composition comprising an antibody of claim 1 in association with at least one pharmaceutically acceptable excipient.

17. A method of treatment of diseases mediated by IL-4 and/or IgE which treatment comprises administering to a subject in need of such treatment an effective amount of an antibody of claim 1.

18. A method of treatment, where in the disease is selected from the group consisting of atopic dermatitis, allergic asthma and allergic rhinitis.