Anti-bcma antibodies, pharmaceutical compositions thereof, and uses

Abstract

The present application provides anti-BCMA antibodies, pharmaceutical compositions thereof, and uses thereof. Sequences of the anti-BCMA antibodies of the present application are described herein. The anti-BCMA antibodies of the present application, or antigen-binding fragments thereof, and pharmaceutical compositions thereof, can be used to treat B-cell related diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to anti-BCMA antibodies, pharmaceutical compositions thereof and uses thereof. BACKGROUND

[0002] B-cell maturation antigen (BCMA), also known as CD269, consists of 184 amino acid residues, and its intracellular region contains 80 amino acid residues. The extracellular region sequence only has one carbohydrate recognition domain for B cell surface molecules. BCMA belongs to the type I transmembrane signaling protein lacking a signal peptide, and is a member of the tumor necrosis factor receptor family. It can be combined with two ligands, B cell activating factor BAFF or proliferation-inducing ligand (APRIL), respectively. In normal tissues, BCMA is expressed on the surface of mature B cells and plasma cells. The immune system of BCMA knockout mice is normal, and the spleen structure is normal. The development of B lymphocytes is normal, but the number of plasma cells is significantly reduced, which proves that BCMA plays an important role in maintaining the survival of plasma cells. The mechanism mainly includes the combination of BCMA with BAFF protein, and the up-regulation of anti-apoptotic genes Bcl-2, Mcl-1 and Bclw, etc., to maintain cell growth. Similarly, this mechanism also plays a function in myeloma cells, and plays an important role in promoting the malignant proliferation of myeloma cells. Studies have shown that BCMA is widely expressed in multiple myeloma cell lines, and consistent results have been obtained in the detection of multiple myeloma patients. On the basis of the existing reports, Kochenderfer et al. further studied the expression characteristics of BCMA by combining Q-PCR, flow cytometry and immunohistochemical methods, and confirmed that BCMA was not expressed in normal human tissues other than mature B cells and plasma cells, and was also not expressed in CD34+ hematopoietic cells. SUMMARY

[0003] The present application provides an anti-BCMA antibody or an antigen-binding fragment thereof, which contains at least one CDR selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9.

[0004] In one or more embodiments, the anti-BCMA antibody contains HCDR1 as shown in SEQ ID NO: 1, HCDR2 as shown in SEQ ID NO: 2, and HCDR3 as shown in SEQ ID NO: 3, 4 or 5, and / or contains LCDR1 as shown in SEQ ID NO: 6 or 7, LCDR2 as shown in SEQ ID NO: 8, and LCDR3 as shown in SEQ ID NO: 9.

[0005] In one or more embodiments, the anti-BCMA antibody contains a HCDR1 as set forth in any one of SEQ ID NOs: 12, 20, 28, 36, 44, 52, 60, 68, 76, 84, 92, or 100, a HCDR2 as set forth in any one of SEQ ID NOs: 13, 21, 29, 37, 45, 53, 61, 69, 77, 85, 93, or 101, and a HCDR3 as set forth in any one of SEQ ID NOs: 14, 22, 30, 38, 46, 54, 62, 70, 78, 86, 94, or 102, and / or a LCDR1 as set forth in any one of SEQ ID NOs: 15, 23, 31, 39, 47, 55, 63, 71, 79, 87, 95, or 103, a LCDR2 as set forth in any one of SEQ ID NOs: 16, 24, 32, 40, 48, 56, 64, 72, 80, 88, 96, or 104, and a LCDR3 as set forth in any one of SEQ ID NOs: 17, 25, 33, 41, 49, 57, 65, 73, 81, 89, 97, or 105.

[0006] In one or more embodiments, the anti-BCMA antibody contains a HCDR1, a HCDR2, and a HCDR3 as set forth in any one of Group A to Group L:

[0007] Group HCDR1 HCDR2 HCDR3 A 12 13 14 B 20 21 22 C 28 29 30 D 36 37 38 E 44 45 46 F 52 53 54 G 60 61 62 H 68 69 70 I 76 77 78 J 84 85 86 K 92 93 94 L 100 101 102

[0008] and / or a LCDR1, a LCDR2, and a LCDR3 as set forth in any one of Group 1 to Group 12:

[0009]

[0010]

[0011] In one or more embodiments, the anti-BCMA antibody contains a HCDR and a LCDR as set forth in any one of Group a to Group 1:

[0012] Group HCDR1 HCDR2 HCDR3 LCDR1 LCDR2 LCDR3 a 12 13 14 15 16 17 b 20 21 22 23 24 25 c 28 29 30 31 32 33 d 36 37 38 39 40 41 e 44 45 46 47 48 49 f 52 53 54 55 56 57 g 60 61 62 63 64 65 h 68 69 70 71 72 73 i 76 77 78 79 80 81 j 84 85 86 87 88 89 k 92 93 94 95 96 97 l 100 101 102 103 104 105

[0013] In one or more embodiments, the FR1 of the anti-BCMA antibody VH is selected from the FR1 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, 20A2, or 23C4, the FR2 is selected from the FR2 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, or 20A2, the FR3 is selected from the FR3 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 18D10, 20A2, 23C4, or 31F5, the FR4 is selected from the FR4 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 20A2, or 31F5; and / or the FR1 of the VL is selected from the FR1 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, or 20A2, the FR2 is selected from the FR2 of antibody 7E11, 8H7, 15A7, 15H6, 20A2, 23C4, or 31F5, the FR3 is selected from the FR3 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 18D10, 20A2, 23C4, 27A7, or 31F5, the FR4 is selected from the FR4 of antibody 7E11, 11B10, 11G1, 15A7, or 18D10.

[0014] In one or more embodiments, the FR regions of the anti-BCMA antibody VH and VL are the FR regions of the VH and VL of any one of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, 20A2, 20A9, 23C4, 27A7, and 31F5.

[0015] In one or more embodiments, the amino acid sequence of the VH of the anti-BCMA antibody is set forth in any one of SEQ ID NOs: 10, 18, 26, 34, 42, 50, 58, 66, 74, 82, 90, and 98, and / or the amino acid sequence of the VL is set forth in any one of SEQ ID NOs: 11, 19, 27, 35, 43, 51, 59, 67, 75, 83, 91, and 99.

[0016] In one or more embodiments, the VH amino acid sequence of the anti-BCMA antibody is set forth in SEQ ID NO: 10, the VL amino acid sequence is set forth in SEQ ID NO: 11; or the VH amino acid sequence is set forth in SEQ ID NO: 18, the VL amino acid sequence is set forth in SEQ ID NO: 19; the VH amino acid sequence is set forth in SEQ ID NO: 26, the VL amino acid sequence is set forth in SEQ ID NO: 27; the VH amino acid sequence is set forth in SEQ ID NO: 34, the VL amino acid sequence is set forth in SEQ ID NO: 35; the VH amino acid sequence is set forth in SEQ ID NO: 42, the VL amino acid sequence is set forth in SEQ ID NO: 43; the VH amino acid sequence is set forth in SEQ ID NO: 50, the VL amino acid sequence is set forth in SEQ ID NO: 51; the VH amino acid sequence is set forth in SEQ ID NO: 58, the VL amino acid sequence is set forth in SEQ ID NO: 59; the VH amino acid sequence is set forth in SEQ ID NO: 66, the VL amino acid sequence is set forth in SEQ ID NO: 67; the VH amino acid sequence is set forth in SEQ ID NO: 74, the VL amino acid sequence is set forth in SEQ ID NO: 75; the VH amino acid sequence is set forth in SEQ ID NO: 82, the VL amino acid sequence is set forth in SEQ ID NO: 83; the VH amino acid sequence is set forth in SEQ ID NO: 90, the VL amino acid sequence is set forth in SEQ ID NO: 91; the VH amino acid sequence is set forth in SEQ ID NO: 98, the VL amino acid sequence is set forth in SEQ ID NO: 99.

[0017] In one or more embodiments, the heavy chain constant region sequence of the anti-BCMA antibody described in any embodiment of the present application is set forth in SEQ ID NO: 106, and / or the light chain constant region sequence is set forth in SEQ ID NO: 107.

[0018] In one or more embodiments, the anti-BCMA antibody described in any embodiment of the present application is a chimeric antibody or a fully human antibody; preferably a fully human antibody.

[0019] The present application also provides a pharmaceutical composition containing the anti-BCMA antibody or antigen-binding fragment thereof described in any embodiment of the present application, and a pharmaceutically acceptable excipient or carrier.

[0020] The present application also provides a nucleic acid molecule selected from: (1) a polynucleotide sequence encoding the anti-BCMA antibody or antigen-binding fragment thereof described in any embodiment of the present application; (2) a complementary sequence of the polynucleotide sequence of (1).

[0021] The present application also provides the use of the anti-BCMA antibody or antigen binding fragment thereof according to any of the embodiments of the present application in the manufacture of a medicament for treating a B-cell related disease; preferably, the B-cell related disease is a B-cell related tumor or an autoimmune disease.

[0022] The present application also provides a method for treating or preventing a B-cell related disease, comprising administering to a patient in need thereof a therapeutically effective amount of the anti-BCMA antibody or antigen binding fragment thereof according to any of the embodiments of the present application, or a pharmaceutical composition comprising the anti-BCMA antibody or antigen binding fragment thereof according to any of the embodiments of the present application. Preferably, the B-cell related disease is a B-cell related tumor or an autoimmune disease. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 Figure 2 shows the results of the enzyme-linked immunoassay for detecting human BCMA specific serum after immunization of the animals of Example 2.

[0024] Figure 2 Figure 5 shows the flow analysis of the binding of the anti-human BCMA monoclonal antibodies to the BCMA expressed on the surface of U266 cells after purification.

[0025] Figure 3 Figure 6 shows the SDS PAGE gel electrophoresis analysis of each anti-human BCMA monoclonal antibody after purification. DETAILED DESCRIPTION

[0026] The practice of the present application will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are within the skill of the art. Such techniques are explained fully in the literature, such as, Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989); Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Animal Cell Culture (R. I. Freshney, ed., 1987); Methods in Enzymology (Academic Press, Inc.); Current Protocols in Molecular Biology (F. M. Ausubel et al., eds., 1987 and periodic updates thereto); PCR: The Polymerase Chain Reaction (Mullis et al., eds., 1994); A Practical Guide to Molecular Cloning (Perbal Bernard V., 1988); Phage Display: A Laboratory Manual (Barbas et al., 2001).

[0027] BCMA

[0028] As used herein, "BCMA" refers to a cell surface receptor that binds BAFF and / or APRIL or a receptor complex comprising BCMA. The NCBI accession number for the amino acid sequence of human BCMA (huBCMA) is Q02223 (GI:313104029). BCMA proteins can also include variants and fragments. The fragments include fragments of the extracellular domain that do not have all or part of the transmembrane, and / or the intracellular domain as well as the extracellular domain. Soluble forms of huBCMA include the extracellular domain or fragments of the extracellular domain that retain the ability to bind BAFF and / or APRIL. "BCMA" also includes post-translational modifications of the BCMA amino acid sequence. Post-translational modifications include, but are not limited to, N- and O-linked glycosylation.

[0029] Normal tissue expression of BCMA is highly restricted to the B cell lineage, expressed mainly in the secondary follicle / germinal centers of the tonsil / lymph node, on plasmablasts and on differentiated plasma cells. BCMA is expressed in malignant plasma cells at relatively higher levels than observed in normal plasma cells, especially highly expressed in multiple myeloma, smoldering myeloma and monoclonal gammopathy of undetermined significance (MGUS) plasma cells. BCMA is an attractive target for the treatment of B cell related malignancies expressing BCMA, as its expression is highly restricted to normal and malignant plasma cells and thus should have minimal off-target tissue toxicity.

[0030] Anti-BCMA antibodies

[0031] The present application provides antibodies that specifically bind to BCMA.

[0032] The term "antibody" herein includes monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region), antibody compositions with polyepitopic specificity, multispecific antibodies (e.g., bispecific antibodies), diabodies, and single-chain molecules, as well as antibody fragments, particularly antigen binding fragments, e.g., Fab, F(ab')2, and Fv). The terms "immunoglobulin" (Ig) and "antibody" are used interchangeably herein.

[0033] The basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) polypeptides and two identical heavy (H) polypeptides. IgM antibodies can include 5 of the basic heterotetramer units along with an additional polypeptide called a J chain, and contain 10 antigen binding sites; IgA antibodies can include 2-5 of the basic 4 chain units and also contain J chains; and some IgG molecules can include J chains as well. In the case of IgGs, the 4 chain unit is about 150,000 Daltons. Each L polypeptide is linked to a H polypeptide by one or more covalent disulfide bonds and the two H polypeptides are linked to each other by disulfide bonds. Each H and L polypeptide also has intrachain disulfide bonds. Each H polypeptide generally has at its N-terminus a variable domain (VH) followed by three constant domains for each of the α and γ chains and four constant domains for μ and ε isotypes. Each L polypeptide has a variable domain at its N-terminus followed by a constant domain at its other end. The VLand VHare paired to each other, and the CLis paired to the first constant domain of the heavy chain (CH1). Particular amino acid residues at the interface between light and heavy chain variable domains are believed to form an interface. A pair of VHand VLtogether form one antigen binding site. See, e.g., Basic and Clinical Immunology, 8th Ed., Daniel P. Sties, Abba I. Terr and Tristram G. Parslow, eds., Appleton & Lange, Norwalk, CT, 1994, p. 71 and Chapter 6, for a description of the structural and functional characteristics of the different classes of antibodies. The light chain from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains. Depending on the amino acid sequences of the constant domains (CH) of their heavy chains, immunoglobulins can be assigned to different classes or isotypes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, having heavy chains designated a, δ, ε, γ, and μ, respectively. The γ and a classes are further divided into subclasses on the basis of minor

[0034] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domains of the heavy or light chain of an antibody. The variable domains of the heavy chain and light chain can be referred to as "VH" and "VL", respectively. These domains are generally the most variable parts of an antibody (relative to other antibodies of the same class) and contain the antigen binding sites.

[0035] The term "variable" refers to the fact that certain segments of the antibody sequences vary widely in antibodies, depending on the antigen against which the antibody is directed. The variable domains mediate antigen binding and define specificity of a particular antibody for a particular antigen. However, the variability is not evenly distributed across the entire domain; it is concentrated in three segments called hypervariable regions (HVRs) in each of the two variable domains (in both the light chain and the heavy chain variable domains). These HVRs are generally referred to using the abbreviations, HCDR1, HCDR2, and HCDR3 for the heavy chain variable domain and LCDR1, LCDR2, and LCDR3 for the light chain variable domain. The more highly conserved portions of variable domains are referred to as framework regions (FRs). The variable domains of the naturally-occurring heavy and light chains each comprise four FR regions, FR1, FR2, FR3, and FR4, connected by three HVRs. The HVRs in each chain are held together in close proximity by the FR regions and, with the HVRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991). Generally, the structure of light chain variable regions is FR1-LCDR1-FR2-LCDR2-FR3-LCDR3-FR4, and the structure of heavy chain variable regions is FR1-HCDR1-FR2-HCDR2-FR3-HCDR3-FR4. The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity.

[0036] The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and / or post-translation modifications (e.g., isomerization, amidation) that can be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In contrast to polyclonal antibody preparations (which typically include different antibodies directed against different determinants (epitopes) of a given antigen), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, the monoclonal antibodies are advantageous in that they are synthesized by the hybridoma culture, which does not be contaminated with other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present application can be made by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler and Milstein, Nature, 256: 495-97 (1975); Hongo et al., Hybridoma, 14(3): 253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 2nded. 1988; Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas, 563-681, Elsevier, N.Y., 1981), recombinant DNA methods (see, e.g., US 4,816,567), phage display technologies (see, e.g., Clackson et al., Nature, 352: 624-628 (1991); Marks et al., J. Mol. Biol., 222: 581-597 (1992); Sidhu et al., J. Mol. Biol., 338(2): 299-310 (2004); Lee et al., J. Mol. Biol., 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA, 101(34): 12467-12472 (2004); and Lee et al. J. Immunol. Methods, 284(1-2): 119-132 (2004)), and technologies for producing human or human-like antibodies in animals that have parts of or entire human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90: 5107-5111 (1993); Jakobovits, Curr. Opin. Biotechnol., 5: 652- 656 (1994); Bruggeman et al., Year in Immunol., 10: 93-101 (1996); Almage et al., J.USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immunol., 7:33 (1993); US 5,545,807; US 5,545,806; US 5,569,825; US 5,625,126; US 5,633,425; and US 5,661,016; Marks et al., Bio / Technology, 10:779-783 (1992); Lonberg et al., Nature, 368:856-859 (1994); Morrison, Nature, 368:812-813 (1994); Fishwild et al., Nature Biotechnol., 14:845-851 (1996); Neuberger, Nature Biotechnol., 14:826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol., 13:65-93 (1995).

[0037] The terms "full-length antibody," "intact antibody," or "whole antibody" are used interchangeably to refer to an antibody in its entirely (as opposed to an antibody fragment). In particular, whole antibodies include those with heavy chains and light chains including an Fc region. The constant domains can be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variant of those. In some instances, whole antibodies can have one or more effector functions.

[0038] "Antibody fragments" comprise a portion of an intact antibody, preferably the antigen binding and / or variable region of the intact antibody. Antibody fragments preferably are antigen binding fragments of antibodies. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (see U.S. Patent 5,641,870, Example 2; Zapata et al., Protein Eng., 8(10): 1057-1062, 1995); single-chain antibody molecules; scFv-Fc fragments; multispecific antibodies formed from antibody fragments; and any fragments capable of increasing the half-life of the antibody by chemical modification or by incorporation into liposomes. Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, and a residual "Fc" fragment, which contains the antibody's epitope recognition site for the Fc receptor. The Fab fragment consists of an entire light chain and the variable region of a heavy chain, and one heavy chain constant region domain. Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen combining site. Pepsin treatment yields a F(ab')2 fragment that roughly corresponds to two disulfide linked Fab fragments having different antigen binding activities and is still capable of cross-linking antigen. Fab' fragments differ from Fab fragments by having an additional few residues at the carboxy terminus of the CH1 domain including the one or more cysteines from the antibody hinge region. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known. The Fc fragment comprises the carboxy-terminal portions of both H chains held together by disulfides. The effector functions of antibodies are determined in the Fc region, which is also the region recognized by Fc receptors (FcR) found on certain types of cells.

[0039] "Fv" is the minimum antibody fragment that contains a complete antigen- recognition and binding site. This fragment consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association. From the folding of these two domains emanate six hypervariable loops (3 loops each from the H and L chain) that contribute most of the antigen- binding specificity and diversity of an antibody. However, even a single variable domain (or half of an Fv comprising only three HVRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0040] "Single-chain Fv" also abbreviated as "sFv" or "scFv" are antibody fragments that comprise the VHand VLdomains of antibody, linked by a polypeptide linker as a genetically fused protein. Optionally, the sFv further comprises a polypeptide linker between the VHand VLdomains that enables the sFv to form the desired structure for antigen binding. For a review of sFv see The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer- Verlag, New York, pp. 269-315 (1994).

[0041] "Chemical modifications" of the fragments include the addition of poly(alkylene) glycols such as polyethylene glycol ("PEGylation"), PEGylated fragments of Fv, scFv, Fab, F(ab')2and Fab', i.e. Fv-PEG, scFv-PEG, Fab-PEG, F(ab')2-PEG and Fab'-PEG. Such fragments have EGFR binding activity.

[0042] Preferably, the antibody fragments, especially the antigen binding fragments, are constituted by or comprise partial sequences of the heavy chain variable region or of the light chain variable region of the antibody from which they are derived, said partial sequences being sufficient to retain the same binding specificity and sufficient affinity as the antibody from which they are derived, for BCMA, preferably at least equal to 1 / 100 of the affinity of the antibody from which they are derived, and in a more preferred manner at least equal to 1 / 10. Such antibody fragments will comprise a minimum of 5 amino acids, preferably 10, 15, 25, 50 and 100 contiguous amino acids of the sequence of the antibody from which they are derived.

[0043] Monoclonal antibodies also include "chimeric" antibodies (immunoglobulins) in which a portion of the heavy and / or light chain is identical with or homologous to a corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to a corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (US 4,816,567; Morrison et al., Proc. Nat. Acad. Sci. USA, 81 : 6851-6855, 1984).

[0044] "Humanized" forms of non-human (e.g., murine) antibodies refer to chimeric antibodies that contain sequence derived from non-human immunoglobulin. Thus, a "humanized antibody" typically refers to a non-human antibody that has variable domain framework regions exchanged for sequences found in human antibodies. Typically in a humanized antibody, the entire antibody (except the CDRs) is encoded by polynucleotides of human origin or is identical to such an antibody except the CDRs, some or all of which are encoded by nucleic acids derived from a non-human organism. The CDRs, some or all of which are encoded by nucleic acids derived from a non-human organism, are grafted onto the beta-sheet framework of a human antibody variable region to produce an antibody whose specificity is determined by the grafted CDRs. The production of such antibodies is described, for example, in WO 92 / 11018; Jones, 1986, Nature, 321 :522-525; Verhoeyen et al., 1988, Science, 239:1534-1536. Humanized antibodies can also be produced using mice with genetically engineered immune systems (see Roque et al., 2004, Biotechnol. Prog., 20:639-654).

[0045] A "human antibody" refers to an antibody having an amino acid sequence corresponding to that of an antibody produced by a human and / or produced using any of the techniques disclosed herein for making human antibodies. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage display libraries. Such techniques are described, e.g., in Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Methods for the production of human monoclonal antibodies are available, as described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(l):86-95 (1991). See also van Dijk and van de Winkel, Modern Path. 5:368-74 (2001). Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e.g., immunized xenomice (see, e.g., US 6,075,181 and 6,150,584, regarding XENOMOUSE™ technology). See also, e.g., Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006), regarding human antibodies produced by the human B-cell hybridoma technology.

[0046] The anti-BCMA antibodies of the present application can also be minibodies. Minibodies are minimized antibody-like proteins comprising a scFv linked to a CH3 domain (Hu et al., 1996, Cancer Res., 56:3055-3061). The anti-BCMA antibodies of the present application can also be domain antibodies, see, e.g., US 6,248,516. Domain antibodies (dAbs) are functional binding domains of antibodies, corresponding to the variable region of either the heavy chain (VH) or light chain (VL) of a human antibody dAB, with a molecular weight of about 13 kDa or one-tenth the size of a complete antibody. dAbs are well expressed in a variety of hosts including bacterial, yeast, and mammalian cell systems. In addition, dAbs are highly stable and retain activity even after being subjected to harsh conditions, such as freeze-drying or heat denaturation. See, e.g., US 6,291,158; US 6,582,915; US 6,593,081; US 6,172,197; US 2004 / 0110941; EP 0368684; US 6,696,245, WO 04 / 058821, WO 04 / 003019, and WO 03 / 002609.

[0047] The HCDR1 of the anti-BCMA antibodies of the present application can comprise GX1TX2X3X4X5X6 (SEQ ID NO: 1), wherein X1is F or Y, X2is F or S, X3is S, D, T, N, or A, X4is Y, D, S, or A, X5is Y, C, or H, and X6is D, A, or Y. In some embodiments, X1is F, X2is F, X3is A or D, X4is D, X5is Y, C, or H, and X6is A. In some embodiments, X1is Y, X2is F, X3is T, X4is S or A, X5is Y, and X6is A or Y. Exemplary amino acid sequences of HCDR1 are set forth in any one of SEQ ID NOs: 12, 20, 28, 36, 44, 52, 60, 68, 76, 84, 92, or 100.

[0048] The HCDR2 of the anti-BCMA antibody of the present application can contain IX1X2X3X4X5X6X7 (SEQ ID NO: 2), wherein X1is S, N or Y; X2is W, T, A or P; X3is N or G; X4is S or N; X5is D, G or V; X6is T, N, S, D or H; X7is I, M or T. In some embodiments, X1is S, X2is W, X3is N, X4is S, X5is D or V, X6is T, N, H or S, X7is I; preferably, in these embodiments, X5is D, X6is H or N. In some embodiments, X1is N, X2is T or A, X3is G, X4is N, X5is G, X6is N, X7is T or I. The amino acid sequence of an exemplary HCDR2 is set forth in any one of SEQ ID NOs: 13, 21, 29, 37, 45, 53, 61, 69, 77, 85, 93 or 101.

[0049] The HCDR3 of the anti-BCMA antibody of the present application can contain ARGGX1X2X3X4X5X6X7YYX8YYMDV (SEQ ID NO: 3), wherein X1is S or R, X2is I or L, X3is T or E, X4is G or L, X5is N or D, X6is I or V, X7is F or Y, X8is Y or F; in some embodiments, X1is R, X2is L, X3is E, X4is L, X5is D, X6is I or V, X7is Y, X8is F. In some embodiments, the HCDR3 of the anti-BCMA antibody of the present application can contain X1X2X3X4X5X6X7FDY (SEQ ID NO: 4), wherein X1is A or T, X2is K, R or T, X3is V or IQ, X4is S, V or A, X5is G, S or A, X6is A or S, X7is V, S, Y or T. In some embodiments, the HCDR3 of the anti-BCMA antibody of the present application can contain AKDIFSPTGDX1Y (SEQ ID NO: 5), wherein X1is G or D. The amino acid sequence of an exemplary HCDR3 is set forth in any one of SEQ ID NOs: 14, 22, 30, 38, 46, 54, 62, 70, 78, 86, 94 or 102.

[0050] The LCDR1 of the anti-BCMA antibody of the present application can contain QX1IX2X3X4 (SEQ ID NO: 6), wherein X1is S or D, X2is H, I, S or R, X3is S, N, S or T, and X4is Y, F or N. In some embodiments, X1is S or D, X2is I, X3is S, X4is S or T, and X5is Y. In some embodiments, the LCDR1 of the anti-BCMA antibody of the present application can contain QSX1X2X3X4X5X6X7X8Y (SEQ ID NO: 7), wherein X1is L, V or F, X2is L or V, X3is H, Y or S, X4is S or SS, X5is N, Q or D, X6is G or N, X7is Y, K or N, and X8is N or T. In some embodiments, X1is L or V, X2is L, X3is H, X4is S or SS, X5is N, Q or D, X6is G or N, X7is K, and X8is N. An exemplary amino acid sequence of LCDR1 is set forth in any one of SEQ ID NOs: 15, 23, 31, 39, 47, 55, 63, 71, 79, 87, 95, or 103.

[0051] The LCDR2 of the anti-BCMA antibody of the present application can contain X1X2S (SEQ ID NO: 8), wherein X1is L, S, W, G, A or K, and X2is G, A or L. In some embodiments, X1is S, W, G or A, and X2is A. An exemplary amino acid sequence of LCDR2 is set forth in any one of SEQ ID NOs: 16, 24, 32, 40, 48, 56, 64, 72, 80, 88, 96, or 104.

[0052] The LCDR3 of the anti-BCMA antibody of the present application can contain X1X2X3X4X5X6X7X8X9 (SEQ ID NO: 9), wherein X1is M, Q or L, X2is Q, G or H, X3is A, S, Y, R or H, X4is L, F, Y, T or N, X5is Q, S, R, I or H, X6is T, I, P, V, W or Y, X7is P or L, X8is Y, F, L or P, and X9is T or I. In some embodiments, X1is Q, X2is Q, X3is S or Y, X4is F, Y or S, X5is S or R, X6is I or P, X7is P or L, X8is Y, L or F, and X9is T. An exemplary amino acid sequence of LCDR3 is set forth in any one of SEQ ID NOs: 17, 25, 33, 41, 49, 57, 65, 73, 81, 89, 97, or 105.

[0053] In some embodiments, the anti-BCMA antibody of the present application contains a HCDR1 as set forth in SEQ ID NO: 1, a HCDR2 as set forth in SEQ ID NO: 2, and a HCDR3 as set forth in SEQ ID NO: 3, 4, or 5, and / or a LCDR1 as set forth in SEQ ID NO: 6 or 7, a LCDR2 as set forth in SEQ ID NO: 8, and a LCDR3 as set forth in SEQ ID NO: 9. Preferably, the anti-BCMA antibody of the present application contains a HCDR1 as set forth in any one of SEQ ID NO: 12, 20, 28, 36, 44, 52, 60, 68, 76, 84, 92, or 100, a HCDR2 as set forth in any one of SEQ ID NO: 13, 21, 29, 37, 45, 53, 61, 69, 77, 85, 93, or 101, and a HCDR3 as set forth in any one of SEQ ID NO: 14, 22, 30, 38, 46, 54, 62, 70, 78, 86, 94, or 102, and / or a LCDR1 as set forth in any one of SEQ ID NO: 15, 23, 31, 39, 47, 55, 63, 71, 79, 87, 95, or 103, a LCDR2 as set forth in any one of SEQ ID NO: 16, 24, 32, 40, 48, 56, 64, 72, 80, 88, 96, or 104, and a LCDR3 as set forth in any one of SEQ ID NO: 17, 25, 33, 41, 49, 57, 65, 73, 81, 89, 97, or 105.

[0054] Further preferably, the anti-BCMA antibody of the present application contains a HCDR1, a HCDR2, and a HCDR3 as set forth in any one of Group A to Group L:

[0055]

[0056]

[0057] and / or a LCDR1, a LCDR2, and a LCDR3 as set forth in any one of Group 1 to Group 12:

[0058] Group LCDR1 LCDR2 LCDR3 1 15 16 17 2 23 24 25 3 31 32 33 4 39 40 41 5 47 48 49 6 55 56 57 7 63 64 65 8 71 72 73 9 79 80 81 10 87 88 89 11 95 96 97 12 103 104 105

[0059] More preferably, the anti-BCMA antibody of the present application contains a HCDR and a LCDR of any one of Group a to Group 1:

[0060]

[0061]

[0062] The FR1 of the VH of the anti-BCMA antibodies of the application can be selected from the FR1 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, 20A2, or 23C4, the FR2 can be selected from the FR2 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, or 20A2, the FR3 can be selected from the FR3 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 18D10, 20A2, 23C4, or 31F5, and the FR4 can be selected from the FR4 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 20A2, or 31F5; and / or the FR1 of the VL can be selected from the FR1 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, or 20A2, the FR2 can be selected from the FR2 of antibody 7E11, 8H7, 15A7, 15H6, 20A2, 23C4, or 31F5, the FR3 can be selected from the FR3 of antibody 7E11, 8H7, 11B10, 11G1, 15A7, 18D10, 20A2, 23C4, 27A7, or 31F5, and the FR4 can be selected from the FR4 of antibody 7E11, 11B10, 11G1, 15A7, or 18D10.

[0063] In preferred embodiments, the FR regions of the VH and VL of the anti-BCMA antibodies of the application are the FR regions of any of the antibodies 7E11, 8H7, 11B10, 11G1, 15A7, 15H6, 18D10, 20A2, 20A9, 23C4, 27A7, and 31F5. Further preferably, the HCDRs of such antibodies are selected from any of the preceding Groups A through L, and the LCDRs are selected from any of the preceding Groups 1 through 12; more preferably, the CDRs of such antibodies are selected from any of the preceding Groups a through 1.

[0064] In some embodiments, the amino acid sequence of the VH of the anti-BCMA antibody of the application is as set forth in any one of SEQ ID NOs: 10, 18, 26, 34, 42, 50, 58, 66, 74, 82, 90, and 98, and / or the amino acid sequence of the VL is as set forth in any one of SEQ ID NOs: 11, 19, 27, 35, 43, 51, 59, 67, 75, 83, 91, and 99. Preferably, the amino acid sequence of the VH of the anti-BCMA antibody of the application is as set forth in SEQ ID NO: 10, the amino acid sequence of the VL is as set forth in SEQ ID NO: 11; or the amino acid sequence of the VH is as set forth in SEQ ID NO: 18, the amino acid sequence of the VL is as set forth in SEQ ID NO: 19; the amino acid sequence of the VH is as set forth in SEQ ID NO: 26, the amino acid sequence of the VL is as set forth in SEQ ID NO: 27; the amino acid sequence of the VH is as set forth in SEQ ID NO: 34, the amino acid sequence of the VL is as set forth in SEQ ID NO: 35; the amino acid sequence of the VH is as set forth in SEQ ID NO: 42, the amino acid sequence of the VL is as set forth in SEQ ID NO: 43; the amino acid sequence of the VH is as set forth in SEQ ID NO: 50, the amino acid sequence of the VL is as set forth in SEQ ID NO: 51; the amino acid sequence of the VH is as set forth in SEQ ID NO: 58, the amino acid sequence of the VL is as set forth in SEQ ID NO: 59; the amino acid sequence of the VH is as set forth in SEQ ID NO: 66, the amino acid sequence of the VL is as set forth in SEQ ID NO: 67; the amino acid sequence of the VH is as set forth in SEQ ID NO: 74, the amino acid sequence of the VL is as set forth in SEQ ID NO: 75; the amino acid sequence of the VH is as set forth in SEQ ID NO: 82, the amino acid sequence of the VL is as set forth in SEQ ID NO: 83; the amino acid sequence of the VH is as set forth in SEQ ID NO: 90, the amino acid sequence of the VL is as set forth in SEQ ID NO: 91; the amino acid sequence of the VH is as set forth in SEQ ID NO: 98, the amino acid sequence of the VL is as set forth in SEQ ID NO: 99.

[0065] In some embodiments, the amino acid sequence of the heavy chain constant region of the antibody of the application is as set forth in SEQ ID NO: 106, and / or the amino acid sequence of the light chain constant region is as set forth in SEQ ID NO: 107.

[0066] The antibody of the application can be a chimeric antibody, a humanized antibody, or a fully human antibody; preferably a fully human antibody. It is understood that the antibody provided in the embodiments of the application is a fully human antibody.

[0067] Without materially affecting the activity of the antibody, one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more) amino acids can be substituted, added and / or deleted from the sequences of the present application by one of skill in the art to obtain variants of the sequences of the antibody or functional fragment thereof. They are all considered to be included in the scope of protection of the present application. Amino acids with similar properties can be substituted as in the FR and / or CDR regions of the variable region. The substitution is preferably a conservative substitution; amino acid residues that can be conservatively substituted are well known in the art. In some embodiments, the sequences of the variants of the present application can have at least 95%, 96%, 97%, 98%, or 99% identity with the sequences from which they are derived. The sequence identity of the sequences of the present application can be measured using sequence analysis software. For example, using the computer program BLAST, especially BLASTP or TBLASTN, with default parameters.

[0068] The anti-BCMA antibodies of the present application can be modified to affect function. The present application includes anti-BCMA antibodies having modified glycosylation patterns. Modifications can be made to remove undesirable glycosylation sites, or to eliminate the fucose component on the oligosaccharide chains to increase the antibody's function of antibody-dependent cellular cytotoxicity (ADCC), or galactosylation modifications can be made to alter complement-dependent cytotoxicity (CDC).

[0069] The anti-BCMA antibodies of the present application can typically have an affinity constant of about 10 -9 to about 10 -13 M.

[0070] The anti-BCMA antibodies of the application can be prepared using methods conventional in the art, such as the hybridoma technique well known in the art. Alternatively, the anti-BCMA antibodies of the application can be expressed in cell lines other than hybridoma cell lines. A suitable mammalian host cell can be transformed with a sequence encoding an antibody of the application. Transformation can be performed using any known method, including, for example, packaging of the polynucleotide in a virus (or viral vector) and transduction of a host cell with the virus (or vector). The transformation procedure used depends on the host to be transformed. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art, including dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide in liposomes, and direct microinjection of the DNA into nuclei. Mammalian cell lines that can be used as hosts for expression are well known in the art, including, but not limited to, a variety of immortalized cell lines available from the American Type Culture Collection (ATCC), including, but not limited to, Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), and the like. Particularly preferred cell lines are selected by determining which cell lines have high expression levels and produce antibodies with the essential BCMA binding properties.

[0071] Polynucleotide sequences encoding anti-BCMA antibodies

[0072] The present application provides nucleic acid molecules comprising polynucleotide sequences encoding the anti-BCMA antibodies of the application. Provided herein are polynucleotide sequences encoding the heavy chain variable region, the light chain variable region, the heavy chain, the light chain, and each CDR.

[0073] The nucleic acid molecules of the application include DNA and RNA in single- and double-stranded form, as well as the complementary sequences of the above. DNA includes, for example, cDNA, genomic DNA, chemically synthesized DNA, PCR-amplified DNA, and combinations thereof. The nucleic acid molecules of the application include combinations of full-length gene or cDNA molecules and fragments thereof. The nucleic acids of the application are preferably derived from human sources, but nucleic acids derived from non-human sources are also encompassed by the application.

[0074] In the present application, an isolated nucleic acid molecule refers to a nucleic acid molecule in a form that is independent of other sequences, or as a component of a larger nucleic acid construct. In a preferred embodiment, the nucleic acid is substantially free of contaminating endogenous material. The nucleic acid molecule is preferably derived from DNA or RNA that has been isolated at least once in substantially pure form and in an amount or concentration that enables its component nucleotide sequences to be identified, manipulated, and recovered by standard biochemical methods. The sequences are preferably provided and / or constructed in open reading frames that are uninterrupted by internal non-translated sequences or introns, typically present in eukaryotic genes. Sequences of non-translated DNA can be present 5' or 3' to the open reading frame, which likewise do not affect manipulation or expression of the coding region.

[0075] The present application also includes nucleic acids that hybridize to nucleic acids encoding anti-BCMA antibodies as described herein under moderately stringent conditions, preferably under highly stringent conditions. Basic parameters affecting the choice of hybridization conditions and guidance on designing appropriate conditions can be found in Sambrook, Fritsch and Maniatis (1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., Chapters 9 and 11; and Current Protocols in Molecular Biology, 1995, Ausubel et al., eds., John Wiley & Sons, Inc., Sections 2.10 and 6.3-6.4).

[0076] As outlined herein, the variants according to the present application are generally prepared by site-specific mutagenesis of nucleotides in the DNA encoding the anti-BCMA antibodies using cassette mutagenesis or PCR mutagenesis or other techniques well known in the art to generate DNA encoding the variants, and thereafter expressing the recombinant DNA in cell culture. However, antigen-binding fragments comprising up to about 100-150 residues can be prepared by in vitro synthesis using established techniques.

[0077] As will be appreciated by those skilled in the art, due to the degeneracy of the genetic code, a very large number of nucleic acids can be made which all encode the anti-BCMA antibodies of the present application or antigen-binding fragments thereof. Thus, given the identification of a particular amino acid sequence, one skilled in the art can make any number of different nucleic acids by simply modifying the sequence of one or more codons in a manner that does not change the amino acid sequence of the encoded protein.

[0078] The present application also provides expression systems and constructs in the form of plasmids, expression vectors, transcription cassettes or expression cassettes comprising at least one polynucleotide as described above. In addition, the present application provides host cells comprising said expression systems or constructs.

[0079] Expression vectors used in any of the host cells generally contain a sequence for plasmid maintenance and a sequence for cloning and expression of foreign nucleotide sequences. The sequences, collectively referred to in certain embodiments as "flanking sequences," generally include one or more of the following nucleotide sequences: a promoter, one or more enhancer sequences, an origin of replication, a transcription termination sequence, a complete intron sequence containing donor and acceptor splice junctions, a sequence encoding a leader sequence for polypeptide secretion, a ribosome binding site, a polyadenylation sequence, a polylinker region for insertion of nucleic acids encoding antibodies to be expressed, and an optional marker element. Each of these sequences is discussed below.

[0080] The vector can optionally contain a "tag" coding sequence, i.e., an oligonucleotide molecule located at the 5' or 3' end of the anti-BCMA antibody coding sequence; the oligonucleotide sequence encodes a polyhistidine (such as 6His) or another "tag" such as FLAG, HA (hemagglutinin influenza virus), or myc, which are present in commercially available antibodies. This tag is typically fused to the polypeptide when the polypeptide is expressed, and can serve as a means for affinity purification or detection of the anti-BCMA antibody from the host cell. Affinity purification can be accomplished, for example, by column chromatography using an antibody to the tag as the affinity matrix. The tag can optionally be removed from the purified anti-BCMA antibody by various means, such as using certain peptidases for cleavage.

[0081] The flanking sequences can be homologous (i.e., from the same species and / or strain as the host cell), heterologous (i.e., from a species other than the host cell species or strain), hybrid (i.e., a combination of flanking sequences from more than one source), synthetic, or native. Likewise, the source of the flanking sequences can be any prokaryotic or eukaryotic organism, any vertebrate or invertebrate organism, or any plant, provided that the flanking sequences function in and are activatable by the host cell machinery.

[0082] The origin of replication is typically part of the commercially available prokaryotic expression vector, and this origin facilitates the amplification of the vector in the host cell. If the vector of choice does not contain the origin of replication site, it can be chemically synthesized based on known sequences and ligated into the vector. For example, the origin of replication from plasmid pBR322 (New England Biolabs, Beverly, MA) is suitable for most gram-negative bacteria, and various viral origins (e.g., SV40, polyoma, adenovirus, vesicular stomatitis virus (VSV), or papilloma virus, such as HPV or BPV) are useful for cloning vectors in mammalian cells. Mammalian expression vectors usually do not require an origin of replication component (e.g., the SV40 origin is often used because it also contains the viral early promoter).

[0083] The transcription termination sequence is typically located 3' to the polypeptide coding region to terminate transcription. Transcription termination sequences in prokaryotic cells usually are a fragment rich in G-C, followed by a poly-T sequence.

[0084] An optional marker gene encodes a protein necessary for the survival and growth of a host cell grown in a selective culture medium. Typical selection marker genes encode (a) resistance to an antibiotic or other toxin, e.g., ampicillin, tetracycline or kanamycin for prokaryotic host cells; (b) complementation of a nutritional deficiency of the cell; or (c) a protein that supplies an essential nutrient not available from complex or defined media. Specific selectable markers are kanamycin resistance genes, ampicillin resistance genes, and tetracycline resistance genes. Advantageously, neomycin resistance genes can also be used for selection in both prokaryotic and eukaryotic host cells.

[0085] A ribosome binding site is usually necessary for the initiation of translation of the rnRNA and is characterized by a Shine-Dalgarno sequence (prokaryotes) or a Kozak sequence (eukaryotes). This element is typically located 3' to the promoter and 5' to the coding sequence of the polypeptide to be expressed.

[0086] The expression and cloning vectors of the application will typically contain a promoter that is recognized by a host organism and is operably linked to the molecule encoding the anti-BCMA antibody. A promoter is a non-transcribed sequence located 5' to the initiation codon of a structural gene (generally within about 100 to 1000 bp) that controls the transcription of the structural gene.

[0087] Suitable promoters for use with yeast hosts are also well known in the art. Advantageously, yeast enhancers are used with yeast promoters. Suitable promoters for use with mammalian host cells are well known in the art and include, but are not limited to, promoters obtained from viruses such as polyoma, fowlpox, adenovirus, such as adenovirus 2 (Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV40). Other suitable mammalian promoters include heterologous mammalian promoters, e.g., heat shock promoters and actin promoters.

[0088] Enhancer sequences can be inserted into the vectors to increase the transcription of DNA encoding the light or heavy chain of the anti-BCMA antibody of the application by higher eukaryotes. Enhancers are cis-acting elements of DNA that act to increase the transcription of a promoter. Enhancers are generally about from 10 to 300 bp in length and can be derived from genes homologous to the gene being expressed or heterologous to the gene being expressed. Enhancers have been found to be both absolutely and relatively position independent, having been found 5' and 3' to the transcriptional unit. Several enhancer sequences are known from mammalian genes, e.g., those of the globin, elastase, albumin, alpha-fetoprotein and insulin genes. However, enhancers from viruses are most commonly used. The SV40 enhancer, the cytomegalovirus early promoter enhancer, the polyoma enhancer and adenovirus enhancers are examples of

[0089] The expression vectors of the present application can be constructed from a starting vector, such as a commercially available vector. Such vectors can or can not contain all of the desired flanking sequences. If one or more of the flanking sequences described herein are not already present in the vector, they can be obtained separately and ligated to the vector. Methods for obtaining each of the flanking sequences are well known to those of skill in the art.

[0090] After the vector has been constructed and the nucleic acid molecule encoding the light chain, heavy chain, or light chain and heavy chain comprising the anti-BCMA antibody has been inserted into the appropriate site of the vector, the completed vector can be inserted into a suitable host cell for amplification and / or polypeptide expression. The expression vector for the anti-BCMA antibody can be transformed into the selected host cell by well-known methods, including transfection, infection, calcium phosphate co-precipitation, electroporation, microinjection, lipofection, DEAE-dextran-mediated transfection, or other known techniques. The method of choice can vary depending, in part, on the type of host cell to be used.

[0091] When the host cell is cultured under appropriate conditions, it synthesizes the anti-BCMA antibody, which can then be collected from the culture medium (if the host cell secretes it into the medium) or directly from the host cell that produced it (if it is not secreted). Suitable host cells are described previously.

[0092] Use of anti-BCMA antibodies for therapeutic purposes

[0093] All aspects of the anti-BCMA antibodies described herein are useful in the preparation of medicaments for the treatment of various conditions and diseases described herein, particularly conditions associated with B cells (particularly memory B cells and plasma B cells) expressing BCMA, and diseases or conditions associated with B cells expressing BCMA. These diseases, particularly conditions, express BCMA at relatively high levels in malignant plasma cells and monoclonal gammopathy of undetermined significance (MGUS) plasma cells. In some embodiments, the conditions and diseases are B-cell related cancers, but are not limited to plasma cell leukemia; plasmacytoma; B-cell prolymphocytic leukemia; hairy cell leukemia; B-cell non-Hodgkin's lymphoma (NHL); acute myeloid leukemia (AML); chronic myeloid leukemia (CML); acute lymphoblastic leukemia (ALL); chronic lymphocytic leukemia (CLL); follicular lymphoma (including follicular non-Hodgkin's lymphoma types); Burkitt's lymphoma (localized Burkitt's lymphoma; sporadic Burkitt's lymphoma); marginal zone lymphoma (mucosa-associated lymphatic tissue; MALT / MALToma; monocytoid B-cell lymphoma; splenic lymphoma with villous lymphocytes); mantle cell lymphoma; large cell lymphoma (diffuse large cell; diffuse mixed cell; immunoblastic lymphoma; primary mediastinal B-cell lymphoma; angiocentric lymphoma-pulmonary B-cell); small lymphocytic lymphoma (SLL); precursor B-lymphoblastic lymphoma; myeloid leukemia (granulocytic; myelogenous; acute myelogenous leukemia; chronic myelogenous leukemia; subacute myelogenous leukemia; myelocytic sarcoma; chloroma; granulocytic sarcoma; acute promyelocytic leukemia; acute myelomonocytic leukemia); Waldenstrom's macroglobulinemia or other B-cell lymphomas.

[0094] In some embodiments, the conditions and diseases are B-cell related autoimmune disorders, including but not limited to systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), multiple sclerosis (MS), Sjogren's disease, immune-mediated thrombocytopenia, hemolytic anemia, bullous pemphigoid, myasthenia gravis, type I diabetes mellitus, Graves' disease, Addison's disease, pemphigus foliaceus, psoriasis, psoriatic arthritis, and ankylosing spondylitis, among others.

[0095] Diagnostic uses, assays, and kits

[0096] The anti-BCMA antibodies of the present application can be used in diagnostic assays, such as binding assays, to detect and / or quantify BCMA expressed in tissues, such as bone marrow, or cells, such as plasma cells. Anti-BCMA antibodies can be used in research to further study the role of BCMA in disease. Anti-BCMA antibodies can be used to further study the role of BCMA in forming homomeric and / or heteromeric receptor complexes and the role of the BCMA receptor complexes in disease.

[0097] Serum levels of BCMA can be prognostic and a new tool to measure tumor burden (Sanchez E et al., Serum B-cell maturation antigen is elevated in multiple myeloma and correlates with disease status and survival, Br J Haematology, 158, 727-38 (2012)). Embodiments of the present application include diagnostic assays and kits to measure soluble BCMA as a potential surrogate for membrane-bound BCMA on tumor cells.

[0098] The anti-BCMA antibodies of the present application can be used for diagnostic purposes to detect, diagnose or monitor diseases and / or conditions associated with BCMA. The present application provides for detecting the presence of BCMA in a sample using classical immunohistological methods known to those skilled in the art (e.g., Tijssen, 1993, Practice and Theory of Enzyme Immunoassays, Vol. 15, R.H. Burdon and P.H. van Knippenberg, eds., Elsevier, Amsterdam; Zola, 1987, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158, CRC Press, Inc.; Jalkanen et al., 1985, J. Cell. Biol., 101 :976-985; Jalkanen et al., 1987, J. Cell Biol., 105:3087-3096). Detection of BCMA can be performed in vivo or in vitro. Examples of methods suitable for detecting the presence of BCMA include ELISA, FACS, RIA, and the like.

[0099] For diagnostic applications, the anti-BCMA antibodies are typically labeled with a detectable labeling group. Suitable labeling groups include, but are not limited to, the following: radioisotopes or radionuclides (e.g., 3 H, 14 C, 15 N, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131I) a fluorescent group (e.g., FITC, rhodamine, lanthanide phosphors), an enzymatic group (e.g., horseradish peroxidase, beta-galactosidase, luciferase, alkaline phosphatase), a chemiluminescent group, a biotinyl group, or a predetermined polypeptide epitope recognized by a secondary reporter (e.g., a leucine zipper pair sequence, a binding site for a secondary antibody, a metal binding domain, an epitope tag). In some embodiments, the labeling group is coupled to the anti-BCMA antibody through a spacer arm of various lengths to reduce potential steric hindrance. Various methods for labeling proteins are known in the art and can be used to practice the present application.

[0100] One aspect of the present application provides for the identification of cells expressing BCMA. In one embodiment, the antibody is labeled with a labeling group and the binding of the labeled antibody to BCMA is detected. In another embodiment, the binding of the antibody to BCMA is detected in vivo. In another embodiment, the antibody-BCMA is isolated and measured using techniques known in the art.

[0101] Another aspect of the present application provides for the detection of the presence of a test molecule that competes with the antibody of the present application for binding to BCMA. One example of such an assay would involve detecting the amount of free antibody in a solution containing a quantity of BCMA in the presence or absence of the test molecule. An increase in the amount of free antibody (i.e., antibody that is not bound to BCMA) would indicate that the test molecule is able to compete with the antibody for binding to BCMA. In one embodiment, the antibody is labeled with a labeling group. Alternatively, the test molecule is labeled and the amount of free test molecule is monitored in the presence or absence of the antibody.

[0102] Pharmaceutical compositions, routes of administration

[0103] The present application provides pharmaceutical compositions comprising a therapeutically effective amount of one or more anti-BCMA antibodies of the present application and a pharmaceutically acceptable diluent, carrier, solubilizer, emulsifier, preservative, and / or adjuvant.

[0104] In certain embodiments, the acceptable diluent, carrier, solubilizer, emulsifier, preservative, and / or adjuvant, etc. in the pharmaceutical composition is preferably nontoxic to recipients at the dosages and concentrations employed. In certain embodiments, the pharmaceutical composition can contain such materials as are known in the art for improving, maintaining or preserving the shelf life of the composition, e.g., pH and ionic strength of the composition. These materials are non-toxic in the amounts and concentrations employed, and include buffers, antioxidants, bulking agents, lubricants, and the like. These materials can be selected from among those that are known to be pharmaceutically acceptable. Remington's Pharmaceutical Sciences, 18th Ed., A. R. Genrmo, ed., Mack Publishing Company, Easton, PA, 1990, can be consulted to determine which materials can be included in the composition. The optimal pharmaceutical composition will be determined by the particular characteristics of the active agent and the intended route of administration.

[0105] The pharmaceutical compositions of the application can be selected for parenteral delivery. Alternatively, the compositions can be selected for inhalation or delivery through the digestive tract, such as orally. The preparation of such pharmaceutically acceptable compositions is within the skill of the art.

[0106] Other pharmaceutical compositions will be apparent to those skilled in the art, including formulations comprising the anti-BCMA antibodies in sustained or controlled release delivery formulations. Techniques for formulating a variety of other sustained or controlled delivery means, such as liposome carriers, bioerodible microparticles or porous beads, and depot injections, are also known to those skilled in the art.

[0107] Pharmaceutical compositions for in vivo administration are typically provided in the form of a sterile preparation. Sterilization is accomplished by filtration through a sterile filtration membrane. This method can be used in sterilizing the composition when it is lyophilized, either prior to or following lyophilization and rehydration. Compositions for parenteral administration can be stored in a lyophilized form or in solution. Parenteral compositions are generally placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0108] Once formulated, the pharmaceutical compositions are stored in unit or multi-dose containers, in solid, crystal, or in dehydrated or lyophilized powder form for solution or suspension, in sterile vials. The formulations can be stored as a ready-to-use form or in a form that is reconstituted prior to administration (e.g., lyophilized). The present application also provides kits for producing single dose administration units. The kits of the present application can each contain a first container having a dry protein and a second container having an aqueous formulation. In certain embodiments of the present application, kits containing single and multiple chamber pre-filled syringes (e.g., liquid syringes and lyophilized syringes) are provided.

[0109] The present application also provides methods of treating a patient, particularly a patient's B-cell related disease, such as a B-cell related cancer and autoimmune disease, by administering an anti-BCMA antibody or antigen binding fragment thereof or a pharmaceutical composition thereof according to any of the embodiments of the present application.

[0110] As used herein, the terms "patient," "subject," "individual," "subject," are used interchangeably herein and include any organism, preferably an animal, more preferably a mammal (e.g., a rat, a mouse, a dog, a cat, a rabbit, etc.), and most preferably a human. "Treatment" means the use of a therapeutic regimen described herein on a subject to achieve at least one positive therapeutic effect (e.g., a reduction in the number of cancer cells, a reduction in tumor size, a reduction in the rate of cancer cell infiltration into peripheral organs, or a reduction in the rate of tumor metastasis or tumor growth). The therapeutic regimen effective to treat a patient can vary depending on a variety of factors, such as the patient's disease state, age, body weight, and the ability of the therapy to elicit an anti-cancer response in the subject.

[0111] The therapeutically effective amount of a pharmaceutical composition containing an anti-BCMA antibody or antigen-binding fragment thereof of the present application to be employed will depend, for example, on the therapy intended and the subject. Those of ordinary skill in the art will appreciate that appropriate dosage levels for treatment will vary depending on the molecule delivered, the indication, the route of administration, and the size (body weight, body surface or organ size) and / or condition (age and general health) of the patient. In certain embodiments, the clinician can titer the dosage and modify the route of administration as required by a particular patient to obtain the optimal therapeutic response.

[0112] The frequency of dosing will depend on the pharmacokinetic parameters of the particular anti-BCMA antibody in the formulation being used. Clinicians will generally administer compositions until a dosage is reached that achieves the desired effect. The compositions can therefore be administered as a single dose, or as two or more doses (which can or can not contain the same amounts of the desired molecule) over time, or by means of an implantation device or catheter as a continuous infusion.

[0113] The route of administration of the pharmaceutical composition is according to known methods, for example by injection, intravenously, intraperitoneally, intra-cerebrally (intra-parenchymally), intra-cerebroventricularly, intramuscularly, intra-ocularly, intra-arterially, portal or intralesionally; by sustained release systems or by implantation devices.

[0114] The present application will be illustrated hereinafter in the manner of specific examples. It is understood that these examples are merely illustrative and are not intended to limit the scope of the present application. The methods and materials used in the examples are conventional in the art, unless otherwise stated.

[0115] Example 1: Immunization of mice to produce monoclonal antibodies against human BCMA

[0116] The 6-week-old immunoglobulin humanized mice AceMouse were immunized with the expression vector pcDNA3.1(+) expressing the full-length human BCMA protein (sequence reference Uniprot database, number Q02223). The fifth booster immunization was performed with a commercial recombinant human BCMA antigenic protein (Fc tag, ref CS79, supplier Novoprotein) and standard Freund's complete adjuvant at a dose of 25 micrograms of human recombinant BCMA protein antigen per mouse.

[0117] Example 2: Enzyme-linked immunoassay of the sera of the immunized mice

[0118] Human recombinant BCMA antigen protein (Novoprotein, Cat# CS79) was diluted with PBS to 0.5 ng / ul, 100 ul / well of antigen was added to Maxisorp 96-well flat-bottom ELISA plate, sealed with plastic wrap and placed at 4°C overnight. The next day, the antigen was discarded, and then washed once with PBS (200 ul / well), and then patted dry on absorbent paper and 200 ul / well of blocking solution (PBS containing 10% fetal bovine serum) was added to each well and incubated at room temperature for 2 hours. After blocking, the blocking solution was discarded and patted dry on absorbent paper. The serum was diluted in a concentration gradient with 100 ul / well of diluent (PBS containing 5% fetal bovine serum) and incubated at room temperature for 1 hour, after which the sample was discarded, washed 3 times with washing solution (PBS containing 0.05% Tween-20), and finally patted dry on absorbent paper after the washing solution was discarded. 100 ul / well of HRP-conjugated goat anti-mouse IgG secondary antibody (final concentration 0.4 ug / ml; manufacturer Biolegend, Cat# 405306) was added and incubated at room temperature for 1 hour, after which the liquid was discarded and washed 5 times with washing solution (200 ul / well), and finally patted dry on absorbent paper after the washing solution was discarded. 50 ul / well of TMB-hydrogen peroxide urea solution (manufacturer Thermo Scientific TM , Cat# 34029) substrate solution was added and incubated at room temperature in the dark for 3-5 minutes, and the reaction was stopped by adding 50 ul / well of 0.25 M sulfuric acid, after which the optical absorbance at 450 nm was detected on a multifunctional enzyme labeler. The results are shown in Figure 2 .

[0119] Example 3: Obtaining hybridoma cells from the spleen cells of immunized mice using electrofusion technology

[0120] Ag8 mouse myeloma cells were thawed in advance; Ag8 mouse myeloma cells were counted on the day of fusion; the spleen of the successfully immunized BCMA mouse was placed in transport medium RPMI1640 and the cells in the spleen were immediately extracted and counted for standby use; the obtained spleen cells and myeloma cells were each added to 10 ml of RPMI 1640 and washed once, and then 4 x 10 7 splenocytes were mixed with 1 x 10 7A mixture of 8 Ag8 myeloma cells; centrifuge at 200G for 5 minutes after mixing, discard the supernatant, wash twice with 10 ml of fusion solution and reserve; centrifuge at 200G for 5 minutes at room temperature, discard the supernatant and resuspend the precipitate with 2.5 ml of electrofusion buffer; prepare a 15 ml centrifuge tube, add 4.8 ml of preheated RPMI 1640 medium; add the mixed cell suspension to the CUY 497P2 electrode which has been sterilized with 75% alcohol and dried, and perform cell electrofusion using the standard operating procedure of the ECFG21 electrofusion instrument (manufacturer NEPAGENE, model ECFG21); after the fusion is completed, aspirate the cells and place them in 4.8 ml of RPMI 1640 medium which has been preheated in the incubator; resuspend the cells with HAT medium, and plate them at a density of 2 x 10 5 Plate 96-well flat-bottom plates with 100 μl / well of the cell suspension; place the 96-well plates in a 37°C incubator and incubate them, observe the cells daily, and on day 10, take the supernatant for ELISA primary screening.

[0121] Example 4: Screening of Anti-Human BCMA Monoclonal Antibodies Using Enzyme-Linked Immunoassay

[0122] Dilute the BCMA antigen (Novoprotein cat: CS79) with PBS to 0.5 ng / μl; add 100 μl / well of the antigen to the Maxisorp 96-well flat-bottom ELISA plate, seal it with plastic wrap, and place it in a 4°C incubator overnight; the next day, discard the antigen, wash once with PBS (200 μl / well), pat dry on absorbent paper, and add 200 μl / well of blocking solution to each well and incubate at room temperature for 2 hours; after the blocking is completed, discard the blocking solution and pat dry on absorbent paper; add 50 μl of the sample (the culture supernatant of the BCMA hybridoma cells obtained in Example 3) to each well, and incubate at room temperature for 1 hour; then discard the sample, wash 3 times with washing solution, pat dry on absorbent paper after the final washing; add 50 μl / well of HRP-conjugated goat anti-mouse IgG secondary antibody diluted with dilution solution, and incubate at room temperature for 1 hour; then discard the solution, wash 5 times with washing solution (200 μl / well), pat dry on absorbent paper after the final washing; add 50 μl / well of TMB-hydrogen peroxide urea substrate solution, and incubate at room temperature in the dark for 3-5 minutes; add 50 μl / well of 0.25 M sulfuric acid to stop the reaction, and then measure the optical absorbance at 450 nm on a multifunctional microplate reader. The results are shown in Table 1 below.

[0123] Table 1

[0124] Clone No. 7E11 8H7 11B10 11G1 15A7 15H6 OD450 2.299 2.706 2.551 2.713 2.117 2.491 Clone No. 18D10 20A9 23C4 27A7 31F5 20A2 OD450 2.402 2.2 2.86 2.588 3.003 1.309

[0125] Note: Except for 20A2, which was diluted 1:100 for ELISA screening, the supernatant of the other clones was diluted 1:50 for ELISA screening.

[0126] Example 5: Analysis of anti-human BCMA antibody binding to U266 cell surface BCMA using flow cytometry

[0127] After centrifugation, the U266 cells were washed once with PBS containing 3% FCS, then resuspended in 1.5 ml PBS containing 3% FCS, and 25 μl of the cells (2.5 x 10 5 ), 75 μl of sample (BCMA hybridoma cell culture supernatant obtained in Example 3) was added to each well, 75 μl of anti-BCMA antibody (clone 19F2, manufacturer Biolegend, final concentration 1 μg / ml) was added to the positive control wells; 75 μl of IgG2a isotype control (clone MG2a-53, manufacturer Biolegend, final concentration 1 μg / ml) was added to well 1 of the negative control wells; 75 μl of PBS containing 3% FCS was added to well 2 and well 3 of the negative control wells, and the plate was incubated at 4°C for 1 hour, then washed twice with PBS containing 3% FCS; after the supernatant was discarded, 50 μl of secondary antibody (manufacturer ebioscience, goat anti-mouse IgG-PE, item number 12-4010-82) at a concentration of 500 ng / μl was added to the sample wells, the positive control wells, well 1 of the negative control wells, and well 2 of the negative control wells; 50 μl of PBS containing 3% FCS was added to well 3 of the negative control wells, then the plate was incubated at 4°C for 30 minutes in the dark, then washed twice with PBS containing 3% FCS, and finally the cells were resuspended in 50 μl of PBS containing 3% FCS and detected using a flow cytometer. The results are shown in Figure 5. Figure 2

[0128] Example 6: Analysis and purification of anti-human BCMA monoclonal antibody from the BCMA hybridoma supernatant obtained in Example 3 using rProtein G agarose matrix magnetic microspheres

[0129] ​8 ml of hybridoma supernatant was obtained from the culture of BCMA hybridoma cells obtained from Example 3, and 200 μl of rProtein G agarose matrix magnetic microsphere suspension (manufacturer Changzhou Tiandiren Biotech Co., Ltd., product code SM004C) was added to the centrifuge tube and uniformly oscillated at room temperature for 1 hour. After washing twice with 5 times the volume of the magnetic beads, 600 μl of eluent was added, and the solution was blown 5 times with a pipette and eluted for 10 minutes at room temperature with the centrifuge tube gently rotating on a rotating mixer. After the solution became clear, the supernatant was collected, and the eluted component was the target antigen. The supernatant was collected in a new centrifuge tube and immediately 60 μl of neutralizing solution was added to adjust the pH of the eluted component to 7.0-8.0. A small amount of purified antibody was diluted to 200 μg / ml, and 10 μl of the antibody solution was added to 2.5 μl of 5x protein loading buffer (non-denaturing) (manufacturer Sangon Biotech, product code C506032), mixed, and then added to the sample well of 12% Tris-Glycine precast gel (manufacturer Sangon Biotech, product code C661102) along with 3 pre-stained protein markers (manufacturer Sangon Biotech, product code C510010). The precast gel was previously placed in an electrophoresis tank filled with Tris-SDS electrophoresis buffer (manufacturer Sangon Biotech, product code C520001). The electrophoresis was performed according to the standard process of the precast gel manufacturer. After electrophoresis, the gel was stained with universal protein staining solution (manufacturer Sangon Biotech, product code C516024) for 2 hours, and then the scanned image was obtained after elution. The results are shown in Figure 3 The remaining purified monoclonal antibody for later functional analysis was stored at -20°C.

[0130] Example 7: Obtaining of candidate antibody sequence

[0131] The candidate hybridoma cells were cultured, and the cells were collected by centrifugation at 1000 rpm, and total RNA was extracted with Trizol. First strand cDNA was synthesized using this as a template, and then the variable region DNA sequence corresponding to the hybridoma cells was amplified using the first strand cDNA as a subsequent template (Jones and Bendig, 1991). In a 50 μl reaction system, 1 μl of cDNA, 5 μl of 10x PCR buffer, 1 μl of each upstream and downstream primer (25 pmol), 1 μl of dNTP, 1 μl of 25 mmol PL MgCl2, 39 μl of H2O, 1 μl of Taq enzyme, 95°C pre-denaturation for 10 min, temperature cycling, and PCR amplification. The reaction conditions were 94°C denaturation for 1 min, 58°C annealing for 1 min, 72°C extension for 15 s, a total of 32 cycles, and then 72°C incubation for 10 min.

[0132] After sequencing the amplification products, the antibody sequences of the candidate hybridomas were obtained as shown below:

[0133] 7E11

[0134] VH (SEQ ID NO: 10):

[0135] EVQLVESGGGLVQPGGSLRLSCAAS GFTFSYYD MHWVRQGTGKGLEWVSG IGTSGDT YYPDSVKGRFTISRENAKNSLNLQMNSLRDGDTAMYYC ARGPYYYNSSGYYSYDALDI WGQGTMVTVTS

[0136] VL (SEQ ID NO: 11):

[0137] DIVMTQSPLSLSVTPGEPASISCRSS QSLLHSNGYNY LDWYLQKPGQSPQLLIY LGS NRASGVPDRFSGSGSGTDFTLKISRVEAEGVGVYYC MQALQTPYT FGQGTKLEIK

[0138] HCDR1: GFTFSYYD (SEQ ID NO: 12)

[0139] HCDR2: IGTSGDT (SEQ ID NO: 13)

[0140] HCDR3: ARGPYYYNSSGYYSYDALDI (SEQ ID NO: 14)

[0141] LCDR1: QSLLHSNGYNY (SEQ ID NO: 15)

[0142] LCDR2: LGS (SEQ ID NO: 16)

[0143] LCDR3: MQALQTPYT (SEQ ID NO: 17)

[0144] 8H7

[0145] VH (SEQ ID NO: 18):

[0146] EVQLVESGGGLVQPGRSLRISCAGS GFTFDDYA MHWVRQAPGKGLEWVSG ISWNSDTIAYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYC AKVSGAVFDY CGQGTQVTVSS

[0147] VL (SEQ ID NO: 19):

[0148] DIQMTQSPSSLSASVKDRVIITCRAS QSIHSY LNWYQQKPGKAPKLLIY SAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSFSIPYT FGQGTKLEIK

[0149] HCDR1: GFTFDDYA (SEQ ID NO: 20)

[0150] HCDR2: ISWNSDTI (SEQ ID NO: 21)

[0151] HCDR3: AKVSGAVFDY (SEQ ID NO: 22)

[0152] LCDR1: QSIHSY (SEQ ID NO: 23)

[0153] LCDR2: SAS (SEQ ID NO: 24)

[0154] LCDR3: QQSFSIPYT (SEQ ID NO: 25)

[0155] 11B10

[0156] VH (SEQ ID NO: 26):

[0157] QVQLVQSGAEVKKPGASVKVSCKAS GYTFTSYA MHWVRQAPGQRLEWMGW INTGNGNT KYSQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYC ARGGSITGNIFYYYYYMDV WGKGTTVTVAS

[0158] VL (SEQ ID NO: 27):

[0159] DIVVTQSPDSLAVSLGERATINCKSS QSFLSSSNNKNY LAWYQQKPGQPPKLLIY WAS TRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC QQYYSIPFTFGPGTKVDIK

[0160] HCDR1: GYTFTSYA (SEQ ID NO: 28)

[0161] HCDR2: INTGNGNT (SEQ ID NO: 29)

[0162] HCDR3: ARGGSITGNIFYYYYYMDV (SEQ ID NO: 30)

[0163] LCDR1: QSFLSSSNNKNY (SEQ ID NO: 31)

[0164] LCDR2: WAS (SEQ ID NO: 32)

[0165] LCDR3: QQYYSIPFT (SEQ ID NO: 33)

[0166] 11G1

[0167] VH (SEQ ID NO: 34):

[0168] EVQLVESGGGLVQPGRSLRLSCEAS GFTFDDYA MHWVRQPPGKGLEWVSG ISWNSDNI GYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYC AKIQSGSSFDY WGQGTLVTVSS

[0169] VL (SEQ ID NO: 35:

[0170] DIQMTQSPSSLSASVGDRVTITCRAS QSIINF LNWYQQKPGKAPKLLIY GAS NLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYCC QQSSSIPLT FGGGTKVEIK

[0171] HCDR1: GFTFDDYA (SEQ ID NO: 36)

[0172] HCDR2: ISWNSDNI (SEQ ID NO: 37)

[0173] HCDR3: AKIQSGSSFDY (SEQ ID NO: 38)

[0174] LCDR1: QSIINF (SEQ ID NO: 39)

[0175] LCDR2: GAS (SEQ ID NO: 40)

[0176] LCDR3: QQSSSIPLT (SEQ ID NO: 41)

[0177] 15A7

[0178] VH (SEQ ID NO: 42):

[0179] EVQLVESGGGLVQPGGSLRLSCEAS GFTFDDCA MHWVRQTPGKGLEWVSG ISWNSDTM GYADSVKGRFIISRDNAKNSLYLQMNSLRVEDTALYHC TRVRAAVFDY WGQGVLVTVSS

[0180] VL (SEQ ID NO: 43):

[0181] DIHMTQSPSSLSASVGDRVTITCRAS QSISSY LNWFQQKPGKAPTVLIY AAS SLQSGVSSRFSGRGSGADFTLTISSLQPEDFASYFC QQSFSPLYI FGQGTKVEIK

[0182] HCDR1: GFTFDDCA (SEQ ID NO: 44)

[0183] HCDR2: ISWNSDTM (SEQ ID NO: 45)

[0184] HCDR3: TRVRAAVFDY (SEQ ID NO: 46)

[0185] LCDR1: QSISSY (SEQ ID NO: 47)

[0186] LCDR2: AAS (SEQ ID NO: 48)

[0187] LCDR3: QQSFSPLYI (SEQ ID NO: 49)

[0188] 15H6

[0189] VH (SEQ ID NO: 50):

[0190] QVQLVQSGAEVKKPGASAKVSCKAS GYTFTSYA MQWVRQAPGQRLEWMGW INAGNGNTKYSQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYC ARGGRLELDIYYYFYYMDV WGKGTTVTVSS

[0191] VL (SEQ ID NO: 51):

[0192] DIVMSQSPDSLAVSLGERTTINCKSS QSVLHSSQNKNY LAWYQQKPGQPPNPLIH WAS TRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC QQYYRVPFT FGPGTKVDIK

[0193] HCDR1: GYTFTSYA (SEQ ID NO: 52)

[0194] HCDR2: INAGNGNT (SEQ ID NO: 53)

[0195] HCDR3: ARGGRLELDIYYYFYYMDV (SEQ ID NO: 54)

[0196] LCDR1: QSVLHSSQNKNY (SEQ ID NO: 55)

[0197] LCDR2: WAS (SEQ ID NO: 56)

[0198] LCDR3: QQYYRVPFT (SEQ ID NO: 57)

[0199] 18D10

[0200] VH (SEQ ID NO: 58):

[0201] EVRLVESGGGLVQPGRSLRLSCAAS GFTSNDYA MHWVRQAPGRGLEWVSG ISWNSDSI GYADSVKGRFTISRDNAKNSLYLQMNSLRTEDTALYYC ATVVSAYFDY WGQGTLVTVSS

[0202] VL (SEQ ID NO: 59):

[0203] DIQMTQSPSSLSASVGDRVTITCRTS QSISTY LNWYQQKPGKAPKLLIY AAS SLKSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQGSYIIPLT FGGGTRVEIK

[0204] HCDR1: GFTSNDYA (SEQ ID NO: 60)

[0205] HCDR2: ISWNSDSI (SEQ ID NO: 61)

[0206] HCDR3: ATVVSAYFDY (SEQ ID NO: 62)

[0207] LCDR1: QSISTY (SEQ ID NO: 63)

[0208] LCDR2: AAS (SEQ ID NO: 64)

[0209] LCDR3: QGSYIIPLT (SEQ ID NO: 65)

[0210] 20A2

[0211] VH (SEQ ID NO: 66):

[0212] QIQLVQSGAEVKKPGASVKVSCKAS GYTFTAYY LHWVRQSPGHGLEWMGR IYPNSGDT NYAQKFQGRVTMTRDTSINTAYMELSRLRSDDTALYYC ARGFNWNYEGGFDI WGQGTMVTVSS

[0213] VL (SEQ ID NO: 67):

[0214] DVVMTQSPLSLSVTLGQPASISCRSG QSLVYSDGNTY LNWFQQRPGQSPRRLIY KLS SRDSGVPDRFSGSGSGTDFTLKISRMEAEDVGVYYC MQRTHWPPT FGQGTKVEIK

[0215] HCDR1: GYTFTAYY (SEQ ID NO: 68)

[0216] HCDR2: IYPNSGDT (SEQ ID NO: 69)

[0217] HCDR3: ARGFNWNYEGGFDI (SEQ ID NO: 70)

[0218] LCDR1: QSLVYSDGNTY (SEQ ID NO: 71)

[0219] LCDR2: KLS (SEQ ID NO: 72)

[0220] LCDR3: MQRTHWPPT (SEQ ID NO: 73)

[0221] 20A9

[0222] VH (SEQ ID NO: 74):

[0223] QVQLVQSGAEVKKPGASAKVSCKAS GYTFTSYA MQWVRQAPGQRLEWMGW INAGNGNI KYSQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARGGRLELDVYYYFYYMDVWGKGTTVTVSS

[0224] VL (SEQ ID NO: 75):

[0225] DIVMSQSPDSLAVSLGERTTINCKSS QSVLHSSQNKNY LAWYQQKPGQPPNPLIH WAS TRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC QQYYRVPFT FGPGTKVDIK

[0226] HCDR1: GYTFTSYA (SEQ ID NO: 76)

[0227] HCDR2: INAGNGNI (SEQ ID NO: 77)

[0228] HCDR3: ARGGRLELDVYYYFYYMDV (SEQ ID NO: 78)

[0229] LCDR1: QSVLHSSQNKNY (SEQ ID NO: 79)

[0230] LCDR2: WAS (SEQ ID NO: 80)

[0231] LCDR3: QQYYRVPFT (SEQ ID NO: 81)

[0232] 23C4

[0233] VH (SEQ ID NO: 82):

[0234] EVQLVESGGGLVQPGRSLRLSCAASGFTFADHA MHWVRQAPGKGLEWVSG ISWNSDHI GYADSVKGRFTISRDNAKNSLYLQMNSLRPEDTALYYC AKDIFSPTGDGY WGQGTLVTVSS

[0235] VL (SEQ ID NO: 83):

[0236] DIQMTQSPSSLSASVGDRVTITCRAS QDIRNN LGWFQQKPGKTPKRLIY AAS SLQSGVPSRFSGSGSGTEFTLIISSLQPEDFATYYC LHHNSYPPT FGQGTKVEIK

[0237] HCDR1: GFTFADHA (SEQ ID NO: 84)

[0238] HCDR2: ISWNSDHI (SEQ ID NO: 85)

[0239] HCDR3: AKDIFSPTGDGY (SEQ ID NO: 86)

[0240] LCDR1: QDIRNN (SEQ ID NO: 87)

[0241] LCDR2: AAS (SEQ ID NO: 88)

[0242] LCDR3: LHHNSYPPT (SEQ ID NO: 89)

[0243] 27A7

[0244] VH (SEQ ID NO: 90):

[0245] EVQLVESGGGLVQPGRSLRLSCAAS GFTFDDHA MHWVRQAPGKGLEWVSG ISWNSVHI GYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYC AKDIFSPTGDDY WGQGTLVTVSS

[0246] VL (SEQ ID NO: 91):

[0247] DIQMTQSPSSLSASVGDRVTITCRAS QDIRNN LGWFQQKPGKTPKRLIY AASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYC LHHNSYPPT FGQGTKVEIK

[0248] HCDR1: GFTFDDHA (SEQ ID NO: 92)

[0249] HCDR2: ISWNSVHI (SEQ ID NO: 93)

[0250] HCDR3: AKDIFSPTGDDY (SEQ ID NO: 94)

[0251] LCDR1: QDIRNN (SEQ ID NO: 95)

[0252] LCDR2: AAS (SEQ ID NO: 96)

[0253] LCDR3: LHHNSYPPT (SEQ ID NO: 97)

[0254] 31F5

[0255] VH (SEQ ID NO: 98):

[0256] EVQLVESGGGLVQPGRSLRLSCAAS GFTFDDYA MHWVRQAPGKGLEWVSG ISWNSDNI AYADSVKGRFTISRDNAENSLYLQMNSLRTEDTAIYYC AKVAAATFDY RGQGTLVTVSS

[0257] VL (SEQ ID NO: 99):

[0258] DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIF AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYSC QQSFSIPFT FGPGTKVDIK

[0259] HCDR1: GFTFDDYA (SEQ ID NO: 100)

[0260] HCDR2: ISWNSDNI (SEQ ID NO: 101)

[0261] HCDR3: AKVAAATFDY (SEQ ID NO: 102)

[0262] LCDR1: QSISSY (SEQ ID NO: 103)

[0263] LCDR2: AAS (SEQ ID NO: 104)

[0264] LCDR3: QQSFSIPFT (SEQ ID NO: 105)

[0265] The heavy chain constant region sequence of the above antibody is:

[0266] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 106);

[0267] The light chain constant region is:

[0268] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 107).

[0269] Example 8: Analysis of the affinity of anti-human BCMA monoclonal antibodies to human BCMA

[0270] The kits, consumables and instruments used in this example were purchased from GE Healthcare; according to the instructions in the kit, the human BCMA antigen (6His tag, item number CC28, supplier Novoprotein) was coupled to the chip surface, then diluted with HBS-EP+ buffer solution, and different concentrations of the antibody to be analyzed were prepared, with 0 concentration point and quality control concentration point; according to the standard setting program of Biacore T200 manufacturer, the flow cell with captured ligand was used as the detection channel, and the flow cell without captured ligand was used as the reference channel, and HBS-EP+ buffer solution was used as the mobile phase, and different concentrations of the antibody to be analyzed were injected; the SPR signal was collected using Biacore T200 Control Software, and then the data were processed using Biacore T200 Evaluation analysis software; finally, the kinetic analysis or steady-state analysis of the obtained kinetic curve was performed, and the Ka, Kd and KD values were calculated, as shown in Table 2 below.

[0271] Table 2

[0272] Clone No. Ka (1 / Ms) Kd (1 / s) KD (pM) Clone No. Ka (1 / Ms) Kd (1 / s) KD (pM) 20A9 5.726E+5 2.342E-5 40.91 11B10 1.061E+6 2.685E-4 253.10 23C4 4.810E+5 7.015E-5 145.80 11G1 4.715E+5 1.627E-5 34.50 27A7 6.181E+6 7.222E-5 11.69 15A7 4.801E+5 6.021E-5 125.40 31F5 5.764E+5 2.781E-5 48.25 15H6 4.828E+5 4.275E-5 88.54 7E11 5.101E+5 2.572E-5 50.42 18D10 5.095E+5 2.431E-5 47.72 8H7 6.122E+5 1.111E-5 18.14 20A2 8.068E+5 5.294E-5 65.62 SEQUENCE LISTING <110> HUNAN HUAKANG HENGJIAN BIOTECHNOLOGY CO., LTD. <120> Anti-BCMA antibody, pharmaceutical composition thereof and application <130> 19A249 <160> 107 <170> SIPOSequenceListing 1.0 <210> 1 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (2)..(2) <223> Xaa is F or Y <220> <221> MUTAGEN <222> (4)..(4) <223> Xaa is F or S <220> <221> MUTAGEN <222> (5)..(5) <223> Xaa is S, D, T, N or A <220> <221> MUTAGEN <222> (2)..(2) <223> Xaa is S, N, or Y <220> <221> MUTAGEN <222> (3)..(3) <223> Xaa is W, T, A, or P <220> <221> MUTAGEN <222> (4)..(4) <223> Xaa is N or G <400> 1 Gly Xaa Thr Xaa Xaa Xaa Xaa Xaa 1 5 <210> 2 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (5)..(5) <223> Xaa is S or N <220> <221> MUTAGEN <222> (6)..(6) <223> Xaa is D, G, or V <220> <221> MUTAGEN <222> (7)..(7) <223> Xaa is D, A, or Y <220> <221> MUTAGEN <222> (8)..(8) <223> Xaa is D, A, or Y <220> <221> MUTAGEN <222> (9)..(9) <223> Xaa is D, A, or Y <220> <221> MUTAGEN <222> (7)..(7) <223> Xaa is T, N, S, D, or H <220> <221> MUTAGEN <222> (8)..(8) <223> Xaa is I, M, or T <400> 2 Ile Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 <210> 3 <211> 19 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (5)..(5) <223> Xaa is S or R <220> <221> MUTAGEN <222> (6)..(6) <223> Xaa is I or L <220> <221> MUTAGEN <222> (7)..(7) <223> Xaa is T or E <220> <221> MUTAGEN <222> (8)..(8) <223> Xaa is G or L <220> <221> MUTAGEN <222> (9)..(9) <223> Xaa is N or D <220> <221> MUTAGEN <222> (10)..(10) <223> Xaa is I or V <220> <221> MUTAGEN <222> (11)..(11) <223> Xaa is F or Y <220> <221> MUTAGEN <222> (14)..(14) <223> Xaa is F or Y <400> 3 Ala Arg Gly Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr Tyr Xaa Tyr Tyr 1 5 10 15 Met Asp Val <210> 4 <211> 10 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (1)..(1) <223> Xaa is A or T <220> <221> MUTAGEN <222> (2)..(2) <223> Xaa is K, R or T <220> <221> MUTAGEN <222> (3)..(3) <223> Xaa is V or IQ <220> <221> MUTAGEN <222> (4)..(4) <223> Xaa is S, V or A <220> <221> MUTAGEN <222> (5)..(5) <223> Xaa is G, S or A <220> <221> MUTAGEN <222> (6)..(6) <223> Xaa is A or S <220> <221> MUTAGEN <222> (7)..(7) <223> Xaa is V, S, Y, or T <400> 4 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Phe Asp Tyr 1 5 10 <210> 5 <211> 12 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (11)..(11) <223> Xaa is G or D <220> <221> UNSURE <222> (11)..(11) <223> The 'Xaa' at position 11 stands for Gin, Arg, Pro, or Leu. <400> 5 Ala Lys Asp He Phe Ser Pro Thr Gly Asp Xaa Tyr 1 5 10 <210> 6 <211> 6 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (2)..(2) <223> Xaa is S or D <220> <221> MUTAGEN <222> (4)..(4) <223> Xaa is H, I, S, or R <220> <221> MUTAGEN <222> (5)..(5) <223> Xaa is S, N, S or T <220> <221> MUTAGEN <222> (6)..(6) <223> Xaa is Y, F or N <400> 6 Gln Xaa lie Xaa Xaa Xaa 1 5 <210> 7 <211> 11 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (3)..(3) <223> Xaa is L, V or F <220> <221> MUTAGEN <222> (4)..(4) <223> Xaa is L or V <220> <221> MUTAGEN <222> (5)..(5) <223> Xaa is H, Y or S <220> <221> MUTAGEN <222> (6)..(6) <223> Xaa is S or SS <220> <221> MUTAGEN <222> (7)..(7) <223> Xaa is N, Q or D <220> <221> MUTAGEN <222> (8)..(8) <223> Xaa is G or N <220> <221> MUTAGEN <222> (9)..(9) <223> Xaa is Y, K or N <220> <221> MUTAGEN <222> (10)..(10) <223> Xaa is N or T <400> 7 Gln Ser Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Tyr 1 5 10 <210> 8 <211> 3 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (1)..(1) <223> Xaa is L, S, W, G, A or K <220> <221> MUTAGEN <222> (2)..(2) <223> Xaa is G, A or L <400> 8 Xaa Xaa Ser 1 <210> 9 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> MUTAGEN <222> (1)..(1) <223> Xaa is M, Q or L <220> <221> MUTAGEN <222> (2)..(2) <223> Xaa is Q, G or H <220> <221> MUTAGEN <222> (3)..(3) <223> Xaa is A, S, Y, R or H <220> <221> MUTAGEN <222> (4)..(4) <223> Xaa is L, F, Y, T or N <220> <221> MUTAGEN <222> (5)..(5) <223> Xaa is Q, S, R, I or H <220> <221> MUTAGEN <222> (6)..(6) <223> Xaa is T, I, P, V, W or Y <220> <221> MUTAGEN <222> (7)..(7) <223> Xaa is P or L <220> <221> MUTAGEN <222> (8)..(8) <223> Xaa is Y, F, L or P <220> <221> MUTAGEN <222> (9)..(9) <223> Xaa is T or I <400> 9 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 <210> 10 <211> 126 <212> PRT <213> Artificial Sequence <400> 10 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Tyr Tyr 20 25 30 Asp Met His Trp Val Arg Gln Gly Thr Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Gly Thr Ser Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Asn Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Asp Gly Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Arg Gly Pro Tyr Tyr Tyr Asn Ser Ser Gly Tyr Tyr Ser Tyr Asp Ala 100 105 110 Leu Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Thr Ser 115 120 125 <210> 11 <211> 112 <212> PRT <213> Artificial Sequence <400> 11 Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 20 25 30 Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Gly Val Gly Val Tyr Tyr Cys Met Gln Ala 85 90 95 Leu Gln Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 12 <211> 8 <212> PRT <213> Artificial Sequence <400> 12 Gly Phe Thr Phe Ser Tyr Tyr Asp 1 5 <210> 13 <211> 7 <212> PRT <213> Artificial Sequence <400> 13 Ile Gly Thr Ser Gly Asp Thr 1 5 <210> 14 <211> 20 <212> PRT <213> Artificial Sequence <400> 14 Ala Arg Gly Pro Tyr Tyr Tyr Asn Ser Ser Gly Tyr Tyr Ser Tyr Asp 1 5 10 15 Ala Leu Asp Ile 20 <210> 15 <211> 11 <212> PRT <213> Artificial Sequence <400> 15 Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr 1 5 10 <210> 16 <211> 3 <212> PRT <213> Artificial Sequence <400> 16 Leu Gly Ser 1 <210> 17 <211> 9 <212> PRT <213> Artificial Sequence <400> 17 Met Gln Ala Leu Gln Thr Pro Tyr Thr 1 5 <210> 18 <211> 117 <212> PRT <213> Artificial Sequence <400> 18 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Ile Ser Cys Ala Gly Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Thr Ile Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Val Ser Gly Ala Val Phe Asp Tyr Cys Gly Gln Gly Thr Gln 100 105 110 Val Thr Val Ser Ser 115 <210> 19 <211> 107 <212> PRT <213> Artificial Sequence <400> 19 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Lys 1 5 10 15 Asp Arg Val Ile Ile Thr Cys Arg Ala Ser Gln Ser Ile His Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gin Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Ser Phe Ser Ile Pro Tyr 85 90 95 Thr Phe Gly Gin Gly Thr Lys Leu Gin Ile Lys 100 105 <210> 20 <211> 8 <212> PRT <213> Artificial Sequence <400> 20 Gly Phe Thr Phe Asp Asp Tyr Ala 1 5 <210> 21 <211> 8 <212> PRT <213> Artificial Sequence <400> 21 Ile Ser Trp Asn Ser Asp Thr Ile 1 5 <210> 22 <211> 10 <212> PRT <213> Artificial Sequence <400> 22 Ala Lys Val Ser Gly Ala Val Phe Asp Tyr 1 5 10 <210> 23 <211> 6 <212> PRT <213> Artificial Sequence <400> 23 Gln Ser lie His Ser Tyr 1 5 <210> 24 <211> 3 <212> PRT <213> Artificial Sequence <400> 24 Ser Ala Ser 1 <210> 25 <211> 9 <212> PRT <213> Artificial Sequence <400> 25 Gln Gln Ser Phe Ser lie Pro Tyr Thr 1 5 <210> 26 <211> 126 <212> PRT <213> Artificial Sequence <400> 26 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp lie Asn Thr Gly Asn Gly Asn Thr Lys Tyr Ser Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Ser Ile Thr Gly Asn Ile Phe Tyr Tyr Tyr Tyr Tyr 100 105 110 Met Asp Val Trp Gly Lys Gly Thr Thr Val Thr Val Ala Ser 115 120 125 <210> 27 <211> 113 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 27 Asp Ile Val Val Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Phe Leu Ser Ser 20 25 30 Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gin Ala Glu Asp Val Ala Val Tyr Tyr Cys Gin Gin 85 90 95 Tyr Tyr Ser Ile Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Asp Ile 100 105 110 Lys <210> 28 <211> 8 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 28 Gly Tyr Thr Phe Thr Ser Tyr Ala 1 5 <210> 29 <211> 8 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 29 Ile Asn Thr Gly Asn Gly Asn Thr 1 5 <210> 30 <211> 19 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 30 Ala Arg Gly Gly Ser Ile Thr Gly Asn Ile Phe Tyr Tyr Tyr Tyr Tyr 1 5 10 15 Met Asp Val <210> 31 <211> 12 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 31 Gln Ser Phe Leu Ser Ser Ser Asn Asn Lys Asn Tyr 1 5 10 <210> 32 <211> 3 <212> PRT <213> Artificial Sequence <400> 32 Trp Ala Ser 1 <210> 33 <211> 9 <212> PRT <213> Artificial Sequence <400> 33 Gln Gln Tyr Tyr Ser Ile Pro Phe Thr 1 5 <210> 34 <211> 118 <212> PRT <213> Artificial Sequence <400> 34 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Asn Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys lie Gin Ser Gly Ser Ser Phe Asp Tyr Trp Gly Gin Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 35 <211> 107 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 35 Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Ser lie lie Asn Phe 20 25 30 Leu Asn Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu lie 35 40 45 Tyr Gly Ala Ser Asn Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Cys Cys Gin Gin Ser Ser Ser lie Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys 100 105 <210> 36 <211> 8 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 36 Gly Phe Thr Phe Asp Asp Tyr Ala 1 5 <210> 37 <211> 8 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 37 Ile Ser Trp Asn Ser Asp Asn lie 1 5 <210> 38 <211> 11 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 38 Ala Lys lie Gin Ser Gly Ser Ser Phe Asp Tyr 1 5 10 <210> 39 <211> 6 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 39 Gln Ser lie lie Asn Phe 1 5 <210> 40 <211> 3 <212> PRT <213> Artificial Sequence <400> 40 Gly Ala Ser 1 <210> 41 <211> 9 <212> PRT <213> Artificial Sequence <400> 41 Gln Gln Ser Ser Ser Ile Pro Leu Thr 1 5 <210> 42 <211> 117 <212> PRT <213> Artificial Sequence <400> 42 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Asp Asp Cys 20 25 30 Ala Met His Trp Val Arg Gln Thr Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Thr Met Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Leu Tyr His Cys 85 90 95 Thr Arg Val Arg Ala Ala Val Phe Asp Tyr Trp Gly Gln Gly Val Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 43 <211> 107 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 43 Asp Ile His Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Thr Val Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Ser Ser Arg Phe Ser Gly 50 55 60 Arg Gly Ser Gly Ala Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Ser Tyr Phe Cys Gln Gln Ser Phe Ser Pro Leu Tyr 85 90 95 Ile Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 44 <211> 8 <212> PRT <213> Artificial Sequence <400> 44 Gly Phe Thr Phe Asp Asp Cys Ala 1 5 <210> 45 <211> 8 <212> PRT <213> Artificial Sequence <400> 45 Ile Ser Trp Asn Ser Asp Thr Met 1 5 <210> 46 <211> 10 <212> PRT <213> Artificial Sequence <400> 46 Thr Arg Val Arg Ala Ala Val Phe Asp Tyr 1 5 10 <210> 47 <211> 6 <212> PRT <213> Artificial Sequence <400> 47 Gln Ser Ile Ser Ser Tyr 1 5 <210> 48 <211> 3 <212> PRT <213> Artificial Sequence <400> 48 Ala Ala Ser 1 <210> 49 <211> 9 <212> PRT <213> Artificial Sequence <400> 49 Gln Gln Ser Phe Ser Pro Leu Tyr Ile 1 5 <210> 50 <211> 126 <212> PRT <213> Artificial Sequence <400> 50 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Ala Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Met Gln Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Ala Gly Asn Gly Asn Thr Lys Tyr Ser Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Arg Leu Glu Leu Asp Ile Tyr Tyr Tyr Phe Tyr Tyr 100 105 110 Met Asp Val Trp Gly Lys Gly Thr Thr Val Thr Val Ser Ser 115 120 125 <210> 51 ​​​​​​​​​​​​​​​​​​​​​​​<211> 113 <212> PRT <213> Artificial Sequence <400> 51 Asp Ile Val Met Ser Gin Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Thr Thr Ile Asn Cys Lys Ser Ser Gin Ser Val Leu His Ser 20 25 30 Ser Gin Asn Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45 Pro Pro Asn Pro Leu Ile His Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gin Ala Glu Asp Val Ala Val Tyr Tyr Cys Gin Gin 85 90 95 Tyr Tyr Arg Val Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Asp Ile 100 105 110 Lys <210> 52 <211> 8 <212> PRT <213> Artificial Sequence <400> 52 Gly Tyr Thr Phe Thr Ser Tyr Ala 1 5 <210> 53 <211> 8 <212> PRT <213> Artificial Sequence <400> 53 Ile Asn Ala Gly Asn Gly Asn Thr 1 5 <210> 54 <211> 19 <212> PRT <213> Artificial Sequence <400> 54 Ala Arg Gly Gly Arg Leu Glu Leu Asp Ile Tyr Tyr Tyr Phe Tyr Tyr 1 5 10 15 Met Asp Val <210> 55 <211> 12 <212> PRT <213> Artificial Sequence <400> 55 Gln Ser Val Leu His Ser Ser Gln Asn Lys Asn Tyr 1 5 10 <210> 56 <211> 3 <212> PRT <213> Artificial Sequence <400> 56 Trp Ala Ser 1 <210> 57 <211> 9 <212> PRT <213> Artificial Sequence <400> 57 Gln Gln Tyr Tyr Arg Val Pro Phe Thr 1 5 <210> 58 <211> 117 <212> PRT <213> Artificial Sequence <400> 58 Glu Val Arg Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Ser Asn Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Ser Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Thr Val Val Ser Ala Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 59 <211> 107 <212> PRT <213> Artificial Sequence <400> 59 Asp Ile Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gin Ser Ile Ser Thr Tyr 20 25 30 Leu Asn Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Lys Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gin Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gin Gly Ser Tyr Ile Ile Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Arg Val Glu Ile Lys 100 105 <210> 60 <211> 8 <212> PRT <213> Artificial Sequence <400> 60 Gly Phe Thr Ser Asn Asp Tyr Ala 1 5 <210> 61 <211> 8 <212> PRT <213> Artificial Sequence <400> 61 Ile Ser Trp Asn Ser Asp Ser Ile 1 5 <210> 62 <211> 10 <212> PRT <213> Artificial Sequence <400> 62 Ala Thr Val Val Ser Ala Tyr Phe Asp Tyr 1 5 10 <210> 63 <211> 6 <212> PRT <213> Artificial Sequence <400> 63 Gln Ser Ile Ser Thr Tyr 1 5 <210> 64 <211> 3 <212> PRT <213> Artificial Sequence <400> 64 Ala Ala Ser 1 <210> 65 <211> 9 <212> PRT <213> Artificial Sequence <400> 65 Gln Gly Ser Tyr Ile Ile Pro Leu Thr 1 5 <210> 66 <211> 121 <212> PRT <213> Artificial Sequence <400> 66 Gln Ile Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ala Tyr 20 25 30 Tyr Leu His Trp Val Arg Gln Ser Pro Gly His Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Tyr Pro Asn Ser Gly Asp Thr Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Arg Gly Phe Asn Trp Asn Tyr Glu Gly Gly Phe Asp Ile Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 67 <211> 112 <212> PRT <213> Artificial Sequence <400> 67 Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Ser Val Thr Leu Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Arg Ser Gly Gln Ser Leu Val Tyr Ser 20 25 30 Asp Gly Asn Thr Tyr Leu Asn Trp Phe Gin Gin Arg Pro Gly Gin Ser 35 40 45 Pro Arg Arg Leu He Tyr Lys Leu Ser Ser Arg Asp Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Met Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gin Arg 85 90 95 Thr His Trp Pro Pro Thr Phe Gly Gin Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 68 <211> 8 <212> PRT <213> Artificial Sequence <400> 68 Gly Tyr Thr Phe Thr Ala Tyr Tyr 1 5 <210> 69 <211> 8 <212> PRT <213> Artificial Sequence <400> 69 Ile Tyr Pro Asn Ser Gly Asp Thr 1 5 <210> 70 <211> 14 <212> PRT <213> Artificial Sequence <400> 70 Ala Arg Gly Phe Asn Trp Asn Tyr Glu Gly Gly Phe Asp Ile 1 5 10 <210> 71 <211> 11 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 71 Gln Ser Leu Val Tyr Ser Asp Gly Asn Thr Tyr 1 5 10 <210> 72 <211> 3 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 72 Lys Leu Ser 1 <210> 73 <211> 9 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 73 Met Gln Arg Thr His Trp Pro Pro Thr 1 5 <210> 74 <211> 126 <212> PRT <213> Artificial Sequence (Artificial Sequence) <400> 74 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Ala Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Met Gin Trp Val Arg Gin Ala Pro Gly Gin Arg Leu Glu Trp Met 35 40 45 Gly Trp He Asn Ala Gly Asn Gly Asn He Lys Tyr Ser Gin Lys Phe 50 55 60 Gln Gly Arg Val Thr He Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Arg Leu Glu Leu Asp Val Tyr Tyr Tyr Phe Tyr Tyr 100 105 110 Met Asp Val Trp Gly Lys Gly Thr Thr Val Thr Val Ser Ser 115 120 125 <210> 75 <211> 113 <212> PRT <213> Artificial Sequence <400> 75 Asp He Val Met Ser Gin Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Thr Thr He Asn Cys Lys Ser Ser Gin Ser Val Leu His Ser 20 25 30 Ser Gin Asn Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45 Pro Pro Asn Pro Leu lie His Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 lie Ser Ser Leu Gin Ala Glu Asp Val Ala Val Tyr Tyr Cys Gin Gin 85 90 95 Tyr Tyr Arg Val Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Asp lie 100 105 110 Lys <210> 76 <211> 8 <212> PRT <213> Artificial Sequence <400> 76 Gly Tyr Thr Phe Thr Ser Tyr Ala 1 5 <210> 77 <211> 8 <212> PRT <213> Artificial Sequence <400> 77 lie Asn Ala Gly Asn Gly Asn lie 1 5 <210> 78 <211> 19 <212> PRT <213> Artificial Sequence <400> 78 Ala Arg Gly Gly Arg Leu Glu Leu Asp Val Tyr Tyr Tyr Phe Tyr Tyr 1 5 10 15 Met Asp Val <210> 79 <211> 12 <212> PRT <213> Artificial Sequence <400> 79 Gln Ser Val Leu His Ser Ser Gln Asn Lys Asn Tyr 1 5 10 <210> 80 <211> 3 <212> PRT <213> Artificial Sequence <400> 80 Trp Ala Ser 1 <210> 81 <211> 9 <212> PRT <213> Artificial Sequence <400> 81 Gln Gln Tyr Tyr Arg Val Pro Phe Thr 1 5 <210> 82 <211> 119 <212> PRT <213> Artificial Sequence <400> 82 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ala Asp His 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly lie Ser Trp Asn Ser Asp His lie Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Asp lie Phe Ser Pro Thr Gly Asp Gly Tyr Trp Gly Gin Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 83 <211> 107 <212> PRT <213> Artificial Sequence <400> 83 Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Asp lie Arg Asn Asn 20 25 30 Leu Gly Trp Phe Gin Gin Lys Pro Gly Lys Thr Pro Lys Arg Leu lie 35 40 45 Tyr Ala Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Ile Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu His His Asn Ser Tyr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 84 <211> 8 <212> PRT <213> Artificial Sequence <400> 84 Gly Phe Thr Phe Ala Asp His Ala 1 5 <210> 85 <211> 8 <212> PRT <213> Artificial Sequence <400> 85 Ile Ser Trp Asn Ser Asp His Ile 1 5 <210> 86 <211> 12 <212> PRT <213> Artificial Sequence <400> 86 Ala Lys Asp Ile Phe Ser Pro Thr Gly Asp Gly Tyr 1 5 10 <210> 87 <211> 6 <212> PRT <213> Artificial Sequence <400> 87 Gln Asp Ile Arg Asn Asn 1 5 <210> 88 <211> 3 <212> PRT <213> Artificial Sequence <400> 88 Ala Ala Ser 1 <210> 89 <211> 9 <212> PRT <213> Artificial Sequence <400> 89 Leu His His Asn Ser Tyr Pro Pro Thr 1 5 <210> 90 <211> 119 <212> PRT <213> Artificial Sequence <400> 90 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp His 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Val His Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr​​​​​​​​​​​​​​​​​​​​​​​​​​​​​ 65 70 75 80 Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Asp Ile Phe Ser Pro Thr Gly Asp Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 91 <211> 107 <212> PRT <213> Artificial Sequence <400> 91 Asp Ile Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gin Asp Ile Arg Asn Asn 20 25 30 Leu Gly Trp Phe Gin Gin Lys Pro Gly Lys Thr Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gin Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu His His Asn Ser Tyr Pro Pro 85 90 95 Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 92 <211> 8 <212> PRT <213> Artificial Sequence <400> 92 Gly Phe Thr Phe Asp Asp His Ala 1 5 <210> 93 <211> 8 <212> PRT <213> Artificial Sequence <400> 93 lie Ser Trp Asn Ser Val His lie 1 5 <210> 94 <211> 12 <212> PRT <213> Artificial Sequence <400> 94 Ala Lys Asp lie Phe Ser Pro Thr Gly Asp Asp Tyr 1 5 10 <210> 95 <211> 6 <212> PRT <213> Artificial Sequence <400> 95 Gln Asp lie Arg Asn Asn 1 5 <210> 96 <211> 3 <212> PRT <213> Artificial Sequence <400> 96 Ala Ala Ser 1 <210> 97 <211> 9 <212> PRT <213> Artificial Sequence <400> 97 Leu His His Asn Ser Tyr Pro Pro Thr 1 5 <210> 98 <211> 117 <212> PRT <213> Artificial Sequence <400> 98 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Asn Ile Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Glu Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Lys Val Ala Ala Ala Thr Phe Asp Tyr Arg Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 99 <211> 107 <212> PRT <213> Artificial Sequence <400> 99 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Phe Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Ser Cys Gln Gln Ser Phe Ser Ile Pro Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 100 <211> 8 <212> PRT <213> Artificial Sequence <400> 100 Gly Phe Thr Phe Asp Asp Tyr Ala 1 5 <210> 101 <211> 8 <212> PRT <213> Artificial Sequence <400> 101 Ile Ser Trp Asn Ser Asp Asn Ile 1 5 <210> 102 <211> 10 <212> PRT <213> Artificial Sequence <400> 102 Ala Lys Val Ala Ala Ala Thr Phe Asp Tyr 1 5 10 <210> 103 <211> 6 <212> PRT <213> Artificial Sequence <400> 103 Gln Ser Ile Ser Ser Tyr 1 5 <210> 104 <211> 3 <212> PRT <213> Artificial Sequence <400> 104 Ala Ala Ser 1 <210> 105 <211> 9 <212> PRT <213> Artificial Sequence <400> 105 Gln Gln Ser Phe Ser Ile Pro Phe Thr 1 5 <210> 106 <211> 330 <212> PRT <213> Artificial Sequence <400> 106 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 107 <211> 107 <212> PRT <213> Artificial Sequence <400> 107 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105

Claims

1. An anti-BCMA antibody or antigen-binding fragment thereof, characterized in that, The anti-BCMA antibody contains: (1) HCDR1 as shown in SEQ ID NO: 68, HCDR2 as shown in SEQ ID NO: 69, HCDR3 as shown in SEQ ID NO: 70, and contains LCDR1 as shown in SEQ ID NO: 71, LCDR2 as shown in SEQ ID NO: 72, and LCDR3 as shown in SEQ ID NO: 73; or (2) HCDR1 as shown in SEQ ID NO: 28, HCDR2 as shown in SEQ ID NO: 29, HCDR3 as shown in SEQ ID NO: 30, and contains LCDR1 as shown in SEQ ID NO: 31, LCDR2 as shown in SEQ ID NO: 32, and LCDR3 as shown in SEQ ID NO: 33; or (3) HCDR1 as shown in SEQ ID NO: 20, HCDR2 as shown in SEQ ID NO: 21, HCDR3 as shown in SEQ ID NO: 22, and contains LCDR1 as shown in SEQ ID NO: 23, LCDR2 as shown in SEQ ID NO: 24, and LCDR3 as shown in SEQ ID NO:

25.

2. The anti-BCMA antibody or antigen-binding fragment thereof of claim 1, wherein, In the anti-BCMA antibody: the VH amino acid sequence is as shown in SEQ ID NO: 66, and the VL amino acid sequence is as shown in SEQ ID NO: 67; or the VH amino acid sequence is as shown in SEQ ID NO: 26, and the VL amino acid sequence is as shown in SEQ ID NO: 27; or the VH amino acid sequence is as shown in SEQ ID NO: 18, and the VL amino acid sequence is as shown in SEQ ID NO:

19.

3. The anti-BCMA antibody or antigen-binding fragment thereof of claim 1 or 2, wherein, The anti-BCMA antibody is a chimeric antibody or a fully human antibody.

4. A pharmaceutical composition, characterized by, The pharmaceutical composition contains the anti-BCMA antibody or antigen binding fragment thereof according to any one of claims 1-3, and a pharmaceutically acceptable excipient.

5. The pharmaceutical composition of claim 4, wherein The pharmaceutically acceptable excipient is a carrier.

6. A nucleic acid molecule selected from: (1) a polynucleotide sequence encoding the anti-BCMA antibody or antigen binding fragment thereof according to any one of claims 1-3; (2) a complement of the polynucleotide sequence of (1).

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