Monoclonal antibody specifically binding to BCMA or antigen binding fragment thereof and use thereof
By developing high-affinity antibodies that specifically bind BCMA, the problem of lack of efficient binding of BCMA in the prior art has been solved, and the potential therapeutic effect on BCMA-mediated diseases has been achieved.
Patent Information
- Application Number
- CN202380045449.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Priority Date
- 2022-06-09
- Filing Date
- 2023-06-11
- Publication Date
- 2025-02-28
AI Technical Summary
In the prior art, monoclonal antibodies that specifically bind BCMA are lacking in high affinity, making it difficult to effectively treat BCMA-mediated diseases.
Antibodies specifically binding to BCMA and with high binding affinity parameters, including specific amino acid sequences of heavy and light chain variable domains were developed.
The efficient binding and identification of BCMA is achieved, with potential effects in the treatment of BCMA-mediated diseases.
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Figure CN119546642A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the fields of biotechnology and medicine, and in particular to a monoclonal antibody or an antigen-binding fragment thereof that specifically binds to BCMA. The present invention also relates to nucleic acids encoding the antibodies, expression vectors, host cells and methods for producing the antibodies, methods for producing antibodies according to the present invention, pharmaceutical compositions comprising antibodies according to the present invention, pharmaceutical compositions comprising antibodies according to the present invention and other therapeutically active compounds, methods for treating BCMA-mediated diseases or conditions, uses of the antibodies or their pharmaceutical compositions for treating BCMA-mediated diseases or conditions, and uses of the antibodies and other therapeutically active compounds for treating BCMA-mediated diseases or conditions. Background Art
[0002] B cell maturation antigens (BCMA, TNFRSF17, and CD269) are members of the tumor necrosis factor (TNF) receptor superfamily. Their natural ligands are B cell activating factor (BAFF; also known as BLyS or TALL-1, TNFSF13B) and proliferation-inducing ligand (APRIL, TNFSF13, CD256), which are involved in regulating various aspects of humoral immunity, B cell development, and homeostasis. However, the binding affinity of APRIL to BCMA is very low (10 -9 Μ) was significantly higher than BAFF (10 -7 Μ)(Yu-Tzu Tai ET ALL,APRIL andBCMA promote human multiple myeloma growth and immunosuppression in the bonemarrow microenvironment,Blood,2016,127,25,pp.3225–3236,https: / / doi.org / 10.1182 / blood-2016-01-691162).
[0003] BCMA is expressed in plasmablasts and plasma cells (plasma cells, PC) at the late stage of B cell differentiation. In multiple myeloma (MM), the expression of BCMA in malignant plasma cells is significantly increased compared with normal cells, and the activation of BCMA supports the growth and survival of plasma cells by activating MEK / ERK, AKT, NFκB, JNK, p38 and Elk-1 (Shih-Feng Cho ET ALL, Targeting B Cell Maturation Antigen (BCMA) in Multiple Myeloma: Potential Uses of BCMA-Based Immunotherapy, Front Immunol, 2018, 9, 1821, https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC6095983).
[0004] BCMA is a promising target for immunotherapeutic products in various oncological diseases, such as leukemia, lymphoma or multiple myeloma, as this antigen is characterized by limited abundance and increased expression on malignant plasma cells compared to normal plasma cells.
[0005] Patent documents WO2012163805, WO2013072406, WO2014089335, and WO2017143069 describe various antibodies against BCMA.
[0006] To date, only one anti-BCMA antibody has been approved for therapeutic use worldwide as part of an antibody conjugated to a drug (Belantamab mafodotin). In connection with the above, there is a need for novel antibodies that specifically bind to BCMA. Summary of the invention
[0007] The authors of the present group have developed antibodies that specifically bind to BCMA and have high affinity parameters for binding to BCMA.
[0008] Definition and general approach
[0009] Unless otherwise defined herein, all technical and scientific terms used in connection with the present invention shall have the same meanings as commonly understood by one of ordinary skill in the art.
[0010] In addition, unless the context otherwise requires, singular terms shall include plural terms and plural terms shall include singular terms. Generally, existing classifications and methods of cell culture, molecular biology, immunology, microbiology, genetics, analytical chemistry, synthetic organic chemistry, medical and pharmaceutical chemistry, and hybridization and chemistry of proteins and nucleic acids described herein are well known to those skilled in the art and widely used in the art. Enzyme reactions and purification methods are performed according to manufacturer's instructions, as is common in the art or as described herein.
[0011] The term "KD" in the present specification refers to an affinity constant (or equilibrium constant), which is calculated from the ratio of Kd to Ka (ie, Kd / Ka) and expressed as a molar concentration (M).
[0012] "Binding affinity" generally refers to the total intensity of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, "binding affinity" refers to the intrinsic (characteristic, true) binding affinity that reflects the 1:1 interaction between binding pairs of members (e.g., antibodies and antigens). The affinity of a molecule X to its binding partner Y can generally be represented by an affinity constant (KD). A preferred Kd value is about 200nM, 150nM, 100nM, 60nM, 50nM, 40nM, 30nM, 20nM, 10nM, 8nM, 6nM, 4nM, 2nM, 1nM or less. Affinity can be measured by common methods known in the art, including those methods described in this specification. Low-affinity antibodies generally bind antigen slowly and tend to dissociate easily, while high-affinity antibodies generally bind antigen faster and tend to remain bound for a longer time. A variety of methods for measuring binding affinity are known in the art, any of which can be used for the purposes of the present invention.
[0013] The term "Kd", "koff" or "kdis" refers to the dissociation rate constant for a specific interaction between a binding molecule and an antigen. The dissociation rate constant koff can be measured using biolayer interferometry, for example using Octet TM system.
[0014] The term "Ka," "kon," or "association rate" refers to the association rate constant.
[0015] The term “R 2 ” refers to the coefficient of determination.
[0016] The term "response" refers to the antibody-antigen binding signal.
[0017] The term "in vitro" refers to a biological entity, biological process or biological reaction under artificial conditions outside the body. For example, cells grown in vitro are understood to be cells grown in an environment outside the body, such as in a test tube, culture flask or microtiter plate.
[0018] The term "ED 50 ”(EC 50 ) (50% effective dose / concentration) refers to the concentration of an agent that produces 50% of the biological effect (which may include cytotoxicity).
[0019] As used in this specification and the appended claims, unless the context requires otherwise, the words "comprise" and "comprising" or variations such as "includes", "including", "containing" or "containing", will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.
[0020] Antibody
[0021] The present invention relates to monoclonal antibodies or antigen-binding fragments thereof that specifically bind to BCMA (B cell maturation antigen; also known as TNFRSF17 and CD269).
[0022] The term "monoclonal antibody" or "mAb" refers to an antibody that is synthesized and isolated as individual clonal populations.
[0023] The antibodies of the present invention are recombinant antibodies.
[0024] The term "recombinant antibody" refers to an antibody expressed in a cell or cell line that contains a nucleotide sequence encoding the antibody, wherein the nucleotide sequence is not associated with the cell in nature.
[0025] In one aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to BCMA, comprising:
[0026] (a) a heavy chain variable domain comprising:
[0027] (i) having the amino acid sequence SX 1 X 2 CDR1 of MS,
[0028] X 1 =S or G;
[0029] X 2 =A, L or V;
[0030] (ii) having the amino acid sequence X 3 YNGGSDRAGX 4 X 5 DSVX 6 CDR2 of G,
[0031] X 3 =G or C;
[0032] X 4 =F or Y;
[0033] X 5 =A or T;
[0034] X 6 =E or K; and
[0035] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7;
[0036] and
[0037] (b) a light chain variable domain comprising:
[0038] (i) having the amino acid sequence X 7 GX 8 X 9 SNIGX 10 X 11 X 12 X 13 VX 14 CDR1,
[0039] X 7 =S or T;
[0040] X 8 =S, G or T;
[0041] X 9 =S, T, I or R;
[0042] X 10 =0 or A;
[0043] X 11 =S, H, N, G or T;
[0044] X 12 =N, R or Y;
[0045] X 13 =T, A, I or D;
[0046] X 14 =N or H;
[0047] (ii) having the amino acid sequence X 15 X 16 X 17 X 18 CDR2 of RPS,
[0048] X15 = N, R, K, S or G;
[0049] X16 = D, N, G, T or H;
[0050] X17 = S, N or T;
[0051] X18 = Q or N;
[0052] and
[0053] (iii) having the amino acid sequence X 19 X 20 WDX 21 X 22 X 23 X 24 X 25 WX 26 CDR3,
[0054] X 19 =A or S;
[0055] X 20 =A, S, T or V;
[0056] X 21 =G, D, H or S;
[0057] X 22 =S, D or R;
[0058] X 23 =L or V;
[0059] X 24 =N, T, R or S;
[0060] X 25 =V, A, G or N;
[0061] X 26 =M, V or L.
[0062] The term "isolated" used to describe various antibodies according to this specification refers to an antibody that has been identified and separated and / or regenerated from a cell or cell culture expressing the antibody. Impurities (contaminating components) from the natural environment are substances that generally interfere with the diagnostic or therapeutic use of the polypeptide, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. An isolated polypeptide is typically prepared by at least one purification step.
[0063] The term "antibody" or "immunoglobulin" (Ig) used in this specification includes complete antibodies. The term "antibody" refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated as VH in this specification) and a heavy chain constant region. Each light chain consists of a light chain variable region (abbreviated as VL in this specification) and a light chain constant region. Preferably, the light chain is a kappa (κ) light chain, and the constant domain CL is preferably Cκ (κ).
[0064] The antibodies according to the present invention may be of any class (e.g., IgA, IgD, IgE, IgG and IgM, preferably IgG) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2, preferably IgG1).
[0065] The VL and VH regions can be further subdivided into highly variable regions called complementarity determining regions (CDRs), interspersed between more conserved regions called framework regions (FRs). Each VH and VL consists of 3 CDRs and 4 FRs, arranged in the following order from amino-terminus to carboxyl-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens.
[0066] The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (C1q) of the classical complement system.
[0067] The term "antigen binding portion" or "antigen binding fragment" of an antibody as used in this specification refers to one or more antibody fragments that retain the ability to specifically bind to an antigen. It has been shown that the antigen binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments included in the term "antigen binding portion" of an antibody include (i) a Fab fragment, a monovalent fragment consisting of VL, VH, CL and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bridge in the hinge region; (iii) a Fd fragment consisting of VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; (v) a dAb-fragment (Ward et al., (1989) Nature 341: 544-546), which consists of a VH / VHH domain. In addition, the two regions of the Fv fragment, VL and VH, are encoded by different genes and can be connected using a synthetic linker using a recombinant method, which can be received as a single protein chain, wherein the VL and VH regions are paired to form a monovalent molecule (called a single-chain Fv (scFv); see, for example, Bird et al. (1988) Science 242: 423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85: 5879-5883). It is assumed that such single-chain molecules are also included in the term "antigen-binding portion" of an antibody. Such antibody fragments are produced using conventional techniques known to those skilled in the art, and these fragments are screened in the same manner as intact antibodies.
[0068] The "Kabat numbering scheme" or "numbering according to Kabat" as used in this application refers to a system for numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the variable regions of antibody heavy and light chains (Kabat et al. Ann. NY Acad. Sci., 190: 382-93 (1971); Kabat et al. Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242 (1991)).
[0069] An antibody that "specifically binds" a target antigen of the present invention refers to an antibody that binds to the antigen with sufficient affinity so that the antibody can be used as a diagnostic and / or therapeutic agent targeting proteins, cells or tissues expressing the antigen, and has little cross-reaction with other proteins.
[0070] The term "specifically binds" to a particular polypeptide or an epitope on a particular target polypeptide can be described, for example, by a molecule having a Kd for the target of at least about 200 nM, or at least about 150 nM, or at least about 100 nM, or at least about 60 nM, or at least about 50 nM, or at least about 40 nM, or at least about 30 nM, or at least about 20 nM, or at least about 10 nM, or at least about 8 nM, or at least about 6 nM, or at least about 4 nM, or at least about 2 nM, or at least about 1 nM, or greater.
[0071] In one embodiment, the term "specific binding" refers to the binding of a molecule to a specific polypeptide or epitope on a specific polypeptide without substantially binding to any other polypeptide or epitope on a polypeptide.
[0072] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3.
[0073] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:5 or SEQ ID NO:6.
[0074] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14 or SEQ ID NO:15.
[0075] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24.
[0076] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, or SEQ ID NO:35.
[0077] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising:
[0078] A CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3;
[0079] A CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6;
[0080] and
[0081] A CDR3 having the amino acid sequence of SEQ ID NO:7.
[0082] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises:
[0083] (i) a heavy chain variable domain comprising:
[0084] A CDR1 having the amino acid sequence of SEQ ID NO: 1,
[0085] CDR2 having the amino acid sequence of SEQ ID NO: 4 and
[0086] a CDR3 having the amino acid sequence of SEQ ID NO: 7;
[0087] (ii) a heavy chain variable domain comprising:
[0088] A CDR1 having the amino acid sequence of SEQ ID NO: 2,
[0089] A CDR2 having the amino acid sequence of SEQ ID NO: 5 and
[0090] a CDR3 having the amino acid sequence of SEQ ID NO: 7;
[0091] (iii) a heavy chain variable domain comprising:
[0092] A CDR1 having the amino acid sequence of SEQ ID NO: 3,
[0093] CDR2 having the amino acid sequence of SEQ ID NO: 4 and
[0094] a CDR3 having the amino acid sequence of SEQ ID NO: 7;
[0095] (iv) a heavy chain variable domain comprising:
[0096] A CDR1 having the amino acid sequence of SEQ ID NO: 2,
[0097] CDR2 having the amino acid sequence of SEQ ID NO: 4 and
[0098] a CDR3 having the amino acid sequence of SEQ ID NO: 7;
[0099] or
[0100] (v) a heavy chain variable domain comprising:
[0101] A CDR1 having the amino acid sequence of SEQ ID NO: 2,
[0102] A CDR2 having the amino acid sequence of SEQ ID NO: 6 and
[0103] A CDR3 having the amino acid sequence of SEQ ID NO:7.
[0104] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising:
[0105] a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15;
[0106] a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24;
[0107] and
[0108] A CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34 or SEQ ID NO:35.
[0109] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises:
[0110] (i) a light chain variable domain comprising:
[0111] A CDR1 having the amino acid sequence of SEQ ID NO: 8, a CDR2 having the amino acid sequence of SEQ ID NO: 16, and a CDR3 having the amino acid sequence of SEQ ID NO: 25; (ii) a light chain variable domain comprising:
[0112] a CDR1 having the amino acid sequence of SEQ ID NO: 9, a CDR2 having the amino acid sequence of SEQ ID NO: 17, and a CDR3 having the amino acid sequence of SEQ ID NO: 26; (iii) a light chain variable domain comprising:
[0113] a CDR1 having the amino acid sequence of SEQ ID NO: 10, a CDR2 having the amino acid sequence of SEQ ID NO: 18, and a CDR3 having the amino acid sequence of SEQ ID NO: 27; (iv) a light chain variable domain comprising:
[0114] a CDR1 having the amino acid sequence of SEQ ID NO: 11, a CDR2 having the amino acid sequence of SEQ ID NO: 19, and a CDR3 having the amino acid sequence of SEQ ID NO: 28; (v) a light chain variable domain comprising:
[0115] a CDR1 having the amino acid sequence of SEQ ID NO: 12, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 29; (vi) a light chain variable domain comprising:
[0116] a CDR1 having the amino acid sequence of SEQ ID NO: 12, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 30; (vii) a light chain variable domain comprising:
[0117] a CDR1 having the amino acid sequence of SEQ ID NO: 10, a CDR2 having the amino acid sequence of SEQ ID NO: 21, and a CDR3 having the amino acid sequence of SEQ ID NO: 31; (viii) a light chain variable domain comprising:
[0118] A CDR1 having the amino acid sequence of SEQ ID NO: 13,
[0119] A CDR2 having the amino acid sequence of SEQ ID NO: 22 and
[0120] a CDR3 having the amino acid sequence of SEQ ID NO: 32;
[0121] (ix) a light chain variable domain comprising:
[0122] a CDR1 having the amino acid sequence of SEQ ID NO: 14,
[0123] A CDR2 having the amino acid sequence of SEQ ID NO: 23 and
[0124] a CDR3 having the amino acid sequence of SEQ ID NO: 33;
[0125] (x) a light chain variable domain comprising:
[0126] A CDR1 having the amino acid sequence of SEQ ID NO: 8,
[0127] A CDR2 having the amino acid sequence of SEQ ID NO: 24 and
[0128] a CDR3 having the amino acid sequence of SEQ ID NO: 34;
[0129] or
[0130] (xi) a light chain variable domain comprising:
[0131] A CDR1 having the amino acid sequence of SEQ ID NO: 15,
[0132] A CDR2 having the amino acid sequence of SEQ ID NO: 17 and
[0133] A CDR3 having the amino acid sequence of SEQ ID NO:35.
[0134] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises:
[0135] (a) a heavy chain variable domain comprising:
[0136] A CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3;
[0137] A CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6;
[0138] and
[0139] a CDR3 having the amino acid sequence of SEQ ID NO: 7; and
[0140] (b) a light chain variable domain comprising:
[0141] a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15;
[0142] a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24;
[0143] and
[0144] A CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34 or SEQ ID NO:35.
[0145] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises:
[0146] (i)(a) a heavy chain variable domain comprising:
[0147] A CDR1 having the amino acid sequence of SEQ ID NO: 1,
[0148] CDR2 having the amino acid sequence of SEQ ID NO: 4 and
[0149] a CDR3 having the amino acid sequence of SEQ ID NO: 7; and
[0150] (b) a light chain variable domain comprising:
[0151] A CDR1 having the amino acid sequence of SEQ ID NO: 8,
[0152] A CDR2 having the amino acid sequence of SEQ ID NO: 16 and
[0153] a CDR3 having the amino acid sequence of SEQ ID NO: 25;
[0154] (ii)(a) a heavy chain variable domain comprising:
[0155] A CDR1 having the amino acid sequence of SEQ ID NO: 2,
[0156] A CDR2 having the amino acid sequence of SEQ ID NO: 5 and
[0157] a CDR3 having the amino acid sequence of SEQ ID NO: 7; and
[0158] (b) a light chain variable domain comprising:
[0159] A CDR1 having the amino acid sequence of SEQ ID NO:9,
[0160] A CDR2 having the amino acid sequence of SEQ ID NO: 17 and
[0161] a CDR3 having the amino acid sequence of SEQ ID NO: 26;
[0162] (iii)(a) a heavy chain variable domain comprising:
[0163] a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 4, and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising:
[0164] a CDR1 having the amino acid sequence of SEQ ID NO: 10, a CDR2 having the amino acid sequence of SEQ ID NO: 18, and a CDR3 having the amino acid sequence of SEQ ID NO: 27; (iv) (a) a heavy chain variable domain comprising:
[0165] a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 4, and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising:
[0166] a CDR1 having the amino acid sequence of SEQ ID NO: 11, a CDR2 having the amino acid sequence of SEQ ID NO: 19, and a CDR3 having the amino acid sequence of SEQ ID NO: 28; (v)(a) a heavy chain variable domain comprising:
[0167] a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 5, and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising:
[0168] a CDR1 having the amino acid sequence of SEQ ID NO: 12, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 29; (vi)(a) a heavy chain variable domain comprising:
[0169] a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 6, and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising:
[0170] a CDR1 having the amino acid sequence of SEQ ID NO: 12, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 30; (vii) (a) a heavy chain variable domain comprising:
[0171] a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 5, and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising:
[0172] a CDR1 having the amino acid sequence of SEQ ID NO: 10, a CDR2 having the amino acid sequence of SEQ ID NO: 21, and a CDR3 having the amino acid sequence of SEQ ID NO: 31; (viii)(a) a heavy chain variable domain comprising:
[0173] a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 5, and a CDR3 having the amino acid sequence of SEQ ID NO: 7, and (b) a light chain variable domain comprising:
[0174] a CDR1 having the amino acid sequence of SEQ ID NO: 13, a CDR2 having the amino acid sequence of SEQ ID NO: 22, and a CDR3 having the amino acid sequence of SEQ ID NO: 32; (ix)(a) a heavy chain variable domain comprising:
[0175] a CDR1 having the amino acid sequence of SEQ ID NO: 2, a CDR2 having the amino acid sequence of SEQ ID NO: 5, and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising:
[0176] A CDR1 having the amino acid sequence of SEQ ID NO: 12, a CDR2 having the amino acid sequence of SEQ ID NO: 20, and a CDR3 having the amino acid sequence of SEQ ID NO: 30; (x)(a) a heavy chain variable domain comprising:
[0177] A CDR1 having the amino acid sequence of SEQ ID NO: 2,
[0178] CDR2 having the amino acid sequence of SEQ ID NO: 4 and
[0179] a CDR3 having the amino acid sequence of SEQ ID NO: 7; and
[0180] (b) a light chain variable domain comprising:
[0181] a CDR1 having the amino acid sequence of SEQ ID NO: 14,
[0182] A CDR2 having the amino acid sequence of SEQ ID NO: 23 and
[0183] a CDR3 having the amino acid sequence of SEQ ID NO: 33;
[0184] (xi)(a) a heavy chain variable domain comprising:
[0185] A CDR1 having the amino acid sequence of SEQ ID NO: 2,
[0186] CDR2 having the amino acid sequence of SEQ ID NO: 4 and
[0187] a CDR3 having the amino acid sequence of SEQ ID NO: 7; and
[0188] (b) a light chain variable domain comprising:
[0189] A CDR1 having the amino acid sequence of SEQ ID NO: 8,
[0190] A CDR2 having the amino acid sequence of SEQ ID NO: 24 and
[0191] a CDR3 having the amino acid sequence of SEQ ID NO: 34;
[0192] or
[0193] (xii)(a) a heavy chain variable domain comprising:
[0194] A CDR1 having the amino acid sequence of SEQ ID NO: 2,
[0195] A CDR2 having the amino acid sequence of SEQ ID NO: 5 and
[0196] a CDR3 having the amino acid sequence of SEQ ID NO: 7; and
[0197] (b) a light chain variable domain comprising:
[0198] A CDR1 having the amino acid sequence of SEQ ID NO: 15,
[0199] A CDR2 having the amino acid sequence of SEQ ID NO: 17 and
[0200] A CDR3 having the amino acid sequence of SEQ ID NO:35.
[0201] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, or SEQ ID NO:43.
[0202] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, or SEQ ID NO:54.
[0203] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises:
[0204] (a) a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42 or SEQ ID NO:43; and
[0205] (b) a light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53 or SEQ ID NO:54.
[0206] In some embodiments of the invention, the isolated monoclonal antibody or antigen-binding fragment thereof comprises:
[0207] (i)(a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 36, and
[0208] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 44;
[0209] (ii) (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 37, and (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 45;
[0210] (iii) (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38, and (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 46;
[0211] (iv) (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 39, and (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 47;
[0212] (v) (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 40, and (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 48;
[0213] (vi) (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:41, and (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:49;
[0214] (vii)(a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:40, and
[0215] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 50;
[0216] (viii)(a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:40, and
[0217] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 51;
[0218] (ix) (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 40, and (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 49;
[0219] (x)(a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:42, and
[0220] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 52;
[0221] (xi) (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:43, and (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:53;
[0222] or
[0223] (xii)(a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:40, and
[0224] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:54.
[0225] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is a full-length IgG antibody.
[0226] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is of the human IgG1, IgG2, IgG3, or IgG4 isotype.
[0227] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is of the human IgG1 isotype.
[0228] In some embodiments of the invention, the isolated monoclonal antibody comprises the mutations L234A and L235A in CH2 according to the EU numbering of antibody amino acids (or L247A and L248A according to Kabat) (Edelman GM et al., Proc. Natl. Acad. Sci. USA 63 (1969) pp. 78-85; Kabat, EA, et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991).
[0229] In some embodiments of the invention, the isolated monoclonal antibody comprises a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61 or SEQ ID NO:62.
[0230] In some embodiments of the invention, the isolated monoclonal antibody comprises a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, or SEQ ID NO:73.
[0231] In some embodiments of the invention, the isolated monoclonal antibody comprises:
[0232] (i)(a) a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61 or SEQ ID NO:62, and
[0233] (b) a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 or SEQ ID NO:73.
[0234] In some embodiments of the invention, the isolated monoclonal antibody comprises:
[0235] (i)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 55, and
[0236] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 63;
[0237] (ii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 56, and
[0238] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 64;
[0239] (iii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 57, and
[0240] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 65;
[0241] (iv) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 58, and
[0242] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 66;
[0243] (v)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 59, and
[0244] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 67;
[0245] (vi)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 60, and
[0246] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 68;
[0247] (vii)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and
[0248] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 69;
[0249] (viii)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and
[0250] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 70;
[0251] (ix)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and
[0252] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 68,
[0253] (x)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 61, and
[0254] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 71;
[0255] (xi)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 62, and
[0256] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 72;
[0257] or
[0258] (xii)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and
[0259] (b) a light chain comprising the amino acid sequence of SEQ ID NO:73.
[0260] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-001.
[0261] Antibody 01-001 contains:
[0262] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 55; and
[0263] (b) a light chain comprising the amino acid sequence of SEQ ID NO:63.
[0264] Antibody 01-001 contains:
[0265] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 36;
[0266] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:44.
[0267] Antibody 01-001 contains:
[0268] (a) a heavy chain variable domain comprising:
[0269] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 1,
[0270] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4,
[0271] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0272] (b) a light chain variable domain comprising:
[0273] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 8,
[0274] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 16,
[0275] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 25 (Kabat).
[0276] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-006.
[0277] Antibody 01-006 contains:
[0278] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 56; and
[0279] (b) a light chain comprising the amino acid sequence of SEQ ID NO:64.
[0280] Antibody 01-006 contains:
[0281] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 37;
[0282] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:45.
[0283] Antibody 01-006 contains:
[0284] (a) a heavy chain variable domain comprising:
[0285] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0286] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5,
[0287] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0288] (b) a light chain variable domain comprising:
[0289] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 9,
[0290] (ii) a CDR2 having the amino acid sequence of SEQ ID NO: 17 (Kabat),
[0291] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 26 (Kabat).
[0292] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-016.
[0293] Antibody 01-016 contains:
[0294] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 57; and
[0295] (b) a light chain comprising the amino acid sequence of SEQ ID NO:65.
[0296] Antibody 01-016 contains:
[0297] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38;
[0298] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:46.
[0299] Antibody 01-016 contains:
[0300] (a) a heavy chain variable domain comprising:
[0301] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 3,
[0302] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4,
[0303] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0304] (b) a light chain variable domain comprising:
[0305] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 10,
[0306] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 18,
[0307] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 27 (Kabat).
[0308] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-021.
[0309] Antibody 01-021 contains:
[0310] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 58; and
[0311] (b) a light chain comprising the amino acid sequence of SEQ ID NO:66.
[0312] Antibody 01-021 contains:
[0313] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 39;
[0314] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:47.
[0315] Antibody 01-021 contains:
[0316] (a) a heavy chain variable domain comprising:
[0317] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0318] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4,
[0319] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0320] (b) a light chain variable domain comprising:
[0321] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 11,
[0322] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 19,
[0323] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 28 (Kabat).
[0324] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-022.
[0325] Antibody 01-022 contains:
[0326] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 59; and
[0327] (b) a light chain comprising the amino acid sequence of SEQ ID NO:67.
[0328] Antibody 01-022 contains:
[0329] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 40;
[0330] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:48.
[0331] Antibody 01-022 contains:
[0332] (a) a heavy chain variable domain comprising:
[0333] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0334] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5,
[0335] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0336] (b) a light chain variable domain comprising:
[0337] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12,
[0338] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 20,
[0339] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 29 (Kabat).
[0340] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-024.
[0341] Antibody 01-024 contains:
[0342] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 60; and
[0343] (b) a light chain comprising the amino acid sequence of SEQ ID NO:68.
[0344] Antibody 01-024 contains:
[0345] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 41;
[0346] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:49.
[0347] Antibody 01-024 contains:
[0348] (a) a heavy chain variable domain comprising:
[0349] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0350] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 6,
[0351] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0352] (b) a light chain variable domain comprising:
[0353] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12,
[0354] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 20,
[0355] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 30 (Kabat).
[0356] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-025.
[0357] Antibody 01-025 contains:
[0358] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 59; and
[0359] (b) a light chain comprising the amino acid sequence of SEQ ID NO:69.
[0360] Antibody 01-025 contains:
[0361] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 40;
[0362] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:50.
[0363] Antibody 01-025 contains:
[0364] (a) a heavy chain variable domain comprising:
[0365] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0366] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5,
[0367] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0368] (b) a light chain variable domain comprising:
[0369] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 10,
[0370] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 21,
[0371] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 31 (Kabat).
[0372] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-026.
[0373] Antibody 01-026 contains:
[0374] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 59; and
[0375] (b) a light chain comprising the amino acid sequence of SEQ ID NO:70.
[0376] Antibody 01-026 contains:
[0377] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 40;
[0378] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:51.
[0379] Antibody 01-026 contains:
[0380] (a) a heavy chain variable domain comprising:
[0381] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0382] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5,
[0383] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0384] (b) a light chain variable domain comprising:
[0385] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 13,
[0386] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 22,
[0387] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 32 (Kabat).
[0388] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-027.
[0389] Antibody 01-027 contains:
[0390] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 59; and
[0391] (b) a light chain comprising the amino acid sequence of SEQ ID NO:68.
[0392] Antibody 01-027 contains:
[0393] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 40;
[0394] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:49.
[0395] Antibody 01-027 contains:
[0396] (a) a heavy chain variable domain comprising:
[0397] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0398] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5,
[0399] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0400] (b) a light chain variable domain comprising:
[0401] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 12,
[0402] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 20,
[0403] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 30 (Kabat).
[0404] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-030.
[0405] Antibody 01-030 contains:
[0406] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 61; and
[0407] (b) a light chain comprising the amino acid sequence of SEQ ID NO:71.
[0408] Antibody 01-030 contains:
[0409] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 42;
[0410] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:52.
[0411] Antibody 01-030 contains:
[0412] (a) a heavy chain variable domain comprising:
[0413] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0414] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4,
[0415] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0416] (b) a light chain variable domain comprising:
[0417] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 14,
[0418] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 23,
[0419] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 33 (Kabat).
[0420] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-033.
[0421] Antibody 01-033 contains:
[0422] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 62; and
[0423] (b) a light chain comprising the amino acid sequence of SEQ ID NO:72.
[0424] Antibody 01-033 contains:
[0425] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 43;
[0426] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:53.
[0427] Antibody 01-033 contains:
[0428] (a) a heavy chain variable domain comprising:
[0429] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0430] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 4,
[0431] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0432] (b) a light chain variable domain comprising:
[0433] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 8,
[0434] (ii) a CDR2 having the amino acid sequence of SEQ ID NO: 24 (Kabat),
[0435] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 34 (Kabat).
[0436] In some embodiments of the invention, the isolated monoclonal antibody that specifically binds BCMA is antibody 01-037.
[0437] Antibody 01-037 contains:
[0438] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 59; and
[0439] (b) a light chain comprising the amino acid sequence of SEQ ID NO:73.
[0440] Antibody 01-037 contains:
[0441] (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 40;
[0442] (b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:54.
[0443] Antibody 01-037 contains:
[0444] (a) a heavy chain variable domain comprising:
[0445] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 2,
[0446] (ii) a CDR2 (Kabat) having the amino acid sequence of SEQ ID NO: 5,
[0447] (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7 (Kabat), and
[0448] (b) a light chain variable domain comprising:
[0449] (i) a CDR1 (Kabat) having the amino acid sequence of SEQ ID NO: 15,
[0450] (ii) a CDR2 having the amino acid sequence of SEQ ID NO: 17 (Kabat),
[0451] (iii) CDR3 having the amino acid sequence of SEQ ID NO: 35 (Kabat).
[0452] Nucleic acid molecules
[0453] In one aspect, the present invention relates to a nucleic acid encoding any one of the above antibodies or antigen-binding fragments thereof that specifically binds to BCMA.
[0454] In any of the embodiments described, the nucleic acid molecule can be isolated.
[0455] The terms "nucleic acid", "nucleic acid sequence", "nucleic acid sequence", "polynucleotide", "oligonucleotide", "polynucleotide sequence" and "nucleotide sequence" used interchangeably in this specification refer to the precise sequence of nucleotides, modified or unmodified, determining fragments or regions of nucleic acid, containing or not containing non-natural nucleotides, and being double-stranded DNA or RNA, single-stranded DNA or RNA, or the transcription product of said DNA.
[0456] Unless otherwise indicated, the term nucleotide sequence encompasses its complementary sequence. Therefore, a nucleic acid having a specific sequence should be understood to include a nucleic acid having its complementary strand of its complementary sequence.
[0457] An "isolated" nucleic acid molecule is a nucleic acid molecule that has been identified and separated from at least one nucleic acid molecule-impurity. An isolated nucleic acid molecule is different from the form or setting in which it is found under natural conditions. Thus, an isolated nucleic acid molecule is different from a nucleic acid molecule that exists in a cell under natural conditions.
[0458] In one aspect, the present invention relates to a nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence selected from SEQ ID NO: 1 to 73. The nucleic acid molecule may also comprise any combination of said nucleotide sequences.
[0459] As will be appreciated by those skilled in the art, due to the redundancy of the genetic code, a variety of different DNA sequences may encode the amino acid sequence of the light chain or heavy chain of the antibody or its fragment (VH, VL, CDR, etc.) according to the present invention. It is within the skill of those skilled in the art to generate these alternative DNA sequences encoding the same amino acid sequence. Such variant DNA sequences are within the scope of the present invention.
[0460] In some embodiments of the invention, the isolated nucleic acid is DNA.
[0461] The nucleic acid molecules of the present invention can be isolated from any source that produces a monoclonal antibody or antigen-binding fragment thereof that specifically binds BCMA. In certain embodiments of the present invention, the nucleic acid molecules of the present invention can be synthesized by chemical synthesis rather than isolated.
[0462] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-001, and comprises a nucleotide sequence having SEQ ID NO:74.
[0463] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-001, and comprises a nucleotide sequence having SEQ ID NO:82.
[0464] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-001, and comprises a nucleotide sequence having SEQ ID NO:93.
[0465] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-001, and comprises a nucleotide sequence having SEQ ID NO:101.
[0466] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-006, and comprises a nucleotide sequence having SEQ ID NO:75.
[0467] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-006, and comprises a nucleotide sequence having SEQ ID NO:83.
[0468] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-006, and comprises a nucleotide sequence having SEQ ID NO:94.
[0469] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-006, and comprises a nucleotide sequence having SEQ ID NO:102.
[0470] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-016, and comprises a nucleotide sequence having SEQ ID NO:76.
[0471] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-016, and comprises a nucleotide sequence having SEQ ID NO:84.
[0472] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the heavy chain amino acid sequence of antibody 01-016, and comprises a nucleotide sequence having SEQ ID NO:95.
[0473] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-016, and comprises a nucleotide sequence having SEQ ID NO:103.
[0474] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-021, and comprises a nucleotide sequence having SEQ ID NO:77.
[0475] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-021, and comprises a nucleotide sequence having SEQ ID NO:85.
[0476] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-021, and comprises a nucleotide sequence having SEQ ID NO:96.
[0477] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-021, and comprises a nucleotide sequence having SEQ ID NO:104.
[0478] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-022, and comprises a nucleotide sequence having SEQ ID NO:78.
[0479] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-022, and comprises a nucleotide sequence having SEQ ID NO:86.
[0480] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-022, and comprises a nucleotide sequence having SEQ ID NO:97.
[0481] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-022, and comprises a nucleotide sequence having SEQ ID NO:105.
[0482] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-024, and comprises a nucleotide sequence having SEQ ID NO:79.
[0483] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-024, and comprises a nucleotide sequence having SEQ ID NO:87.
[0484] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-024, and comprises a nucleotide sequence having SEQ ID NO:98.
[0485] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-024, and comprises a nucleotide sequence having SEQ ID NO:106.
[0486] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-025, and comprises a nucleotide sequence having SEQ ID NO:78.
[0487] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-025, and comprises a nucleotide sequence having SEQ ID NO:88.
[0488] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-025, and comprises a nucleotide sequence having SEQ ID NO:97.
[0489] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-025, and comprises a nucleotide sequence having SEQ ID NO:107.
[0490] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-026, and comprises a nucleotide sequence having SEQ ID NO:78.
[0491] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-026, and comprises a nucleotide sequence having SEQ ID NO:89.
[0492] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-026, and comprises a nucleotide sequence having SEQ ID NO:97.
[0493] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-026, and comprises a nucleotide sequence having SEQ ID NO:108.
[0494] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-027, and comprises a nucleotide sequence having SEQ ID NO:78.
[0495] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-027, and comprises a nucleotide sequence having SEQ ID NO:87.
[0496] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-027, and comprises a nucleotide sequence having SEQ ID NO:97.
[0497] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-027, and comprises a nucleotide sequence having SEQ ID NO:106.
[0498] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-030, and comprises a nucleotide sequence having SEQ ID NO:80.
[0499] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-030, and comprises a nucleotide sequence having SEQ ID NO:90.
[0500] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-030, and comprises a nucleotide sequence having SEQ ID NO:99.
[0501] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-030, and comprises a nucleotide sequence having SEQ ID NO:109.
[0502] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-033, and comprises a nucleotide sequence having SEQ ID NO:81.
[0503] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-033, and comprises a nucleotide sequence having SEQ ID NO:91.
[0504] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-033, and comprises a nucleotide sequence having SEQ ID NO:100.
[0505] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-033, and comprises a nucleotide sequence having SEQ ID NO:110.
[0506] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain variable domain of antibody 01-037, and comprises a nucleotide sequence having SEQ ID NO:78.
[0507] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain variable domain of antibody 01-037, and comprises a nucleotide sequence having SEQ ID NO:92.
[0508] In some embodiments of the invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the heavy chain of antibody 01-037, and comprises a nucleotide sequence having SEQ ID NO:97.
[0509] In some embodiments of the present invention, the nucleic acid is a nucleic acid encoding the amino acid sequence of the light chain of antibody 01-037, and comprises a nucleotide sequence having SEQ ID NO:111.
[0510] The nucleic acid molecules can be used to express a monoclonal antibody or antigen-binding fragment thereof that specifically binds to BCMA.
[0511] Carrier
[0512] On the one hand, the present invention relates to an expression vector comprising any of the above-mentioned nucleic acid molecules, which encodes the corresponding amino acid sequence of an antibody or a portion thereof (e.g., a heavy chain and / or light chain binding domain sequence) that specifically binds to BCMA. The present invention relates to a vector suitable for expressing any of the nucleotide sequences described herein.
[0513] As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
[0514] As used in this specification, the term "expression" is defined as the transcription and / or translation of a specific nucleotide sequence driven by its promoter.
[0515] In some embodiments of the invention, the vector is a plasmid, a circular double-stranded DNA fragment into which additional DNA segments can be inserted.
[0516] In some embodiments of the invention, the vector is a viral (expression) vector, wherein additional DNA segments may be inserted into the viral genome.
[0517] In some embodiments of the present invention, carriers can replicate autonomously in the host cells into which they are introduced (for example, bacterial vectors and episomal vectors with bacterial replication origin sites). In further embodiments of the present invention, carriers (for example non-episomal vectors) can be integrated into the genome of the host cell after introducing the host cell, and replicated together with the host gene thus. In addition, some carriers can instruct the expression of the genes to which they are operably connected. Such carriers are referred to herein as "recombinant expression vectors" (or simply "expression vectors").
[0518] In some embodiments of the present invention, expression vectors include plasmids, retroviruses, adenoviruses, adeno-associated viruses (AAV), plant viruses such as cauliflower mosaic virus, tobacco mosaic virus, cosmids, YACs, etc. The DNA molecule can be inserted into the vector so that the transcription and translation control sequences in the vector play their intended functions of regulating DNA transcription and translation. The expression vector and expression control sequences can be selected to be compatible with the expression host cell used.
[0519] In one embodiment of the invention, DNA molecules partially or completely encoding the sequences of the first and second binding domains (eg, heavy and light chain sequences, wherein the binding domains comprise heavy and light chain sequences) can be introduced into separate vectors.
[0520] In one embodiment, any combination of the above-mentioned DNA molecules are introduced into the same expression vector.
[0521] In one embodiment of the invention, the DNA molecule can be introduced into the expression vector by standard methods (eg, ligation of complementary restriction sites on the gene segments of the antibody and vector, or blunt end ligation if no restriction sites are present).
[0522] In some embodiments of the present invention, suitable vectors are vectors comprising restriction sites so that any VH or VL sequence can be easily inserted and expressed, as described above. Polyadenylation and transcription termination may occur at the natural chromosomal site downstream of the coding region. The recombinant expression vector can also encode a signal peptide that promotes secretion of the antibody chain from the host cell. The antibody chain gene can be cloned into a vector so that the signal peptide is connected to the amino terminus of the immunoglobulin chain in frame. The signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide from a non-immunoglobulin protein).
[0523] In some embodiments of the present invention, the vector may include an expression control sequence. The term "expression control sequence" used in this specification refers to a polynucleotide sequence necessary for achieving expression and processing of the coding sequence into which it is inserted. It will be appreciated by those skilled in the art that the design of the expression vector, including the selection of expression control sequences, may depend on factors such as the selection of the host cell type to be transformed, the desired antibody expression level, and the like. Expression control sequences include appropriate transcription initiation, termination, promoter, and enhancer sequences; effective RNA processing signals, such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that improve translation efficiency (i.e., Kozak consensus sequences); sequences that enhance protein stability; and sequences that enhance protein secretion when necessary. The nature of such expression control sequences varies depending on the host organism; in prokaryotes, such expression control sequences typically include promoters, ribosome binding sites, and transcription termination sequences; in eukaryotes, such expression control sequences typically include promoters and transcription termination sequences. Preferred expression control sequences for mammalian expression host cells include viral elements that ensure high levels of protein expression in mammalian cells, such as promoters and / or enhancers derived from retroviral LTRs, cytomegalovirus (CMV) (e.g., CMV promoter / enhancer), simian virus 40 (SV40) (e.g., SV40 promoter / enhancer), adenovirus (e.g., major late promoter adenovirus (AdMLP)), polyoma virus, and strong mammalian promoters such as TTR promoter, natural immunoglobulin promoter, or actin promoter. Expression control sequences at least encompass all components whose presence is important for expression and processing.
[0524] In some embodiments of the present invention, in addition to the antibody chain gene and the expression control sequence, the recombinant expression vector of the present invention may also carry additional sequences, such as sequences that regulate the replication of the vector in the host cell (e.g., an origin of replication) and selectable marker genes. The selectable marker gene is useful for selecting host cells into which the vector has been introduced.
[0525] Host cells
[0526] In one aspect, the present invention relates to a method for producing a host cell to produce any of the above antibodies or antigen-binding fragments thereof that specifically bind to BCMA, and comprises transforming the cell with the above vector.
[0527] In one aspect, the present invention relates to a host cell for producing any of the above-mentioned antibodies or antigen-binding fragments thereof that specifically bind to BCMA, comprising any of the above-mentioned nucleic acids.
[0528] The term "host cell" as used herein refers to a cell into which a recombinant expression vector has been introduced. The present invention relates to a host cell, which may comprise, for example, the vector according to the present invention as described above. The present invention also relates to a host cell, which comprises, for example, a nucleotide sequence encoding a heavy chain or an antigen binding portion thereof, a nucleotide sequence encoding a light chain or an antigen binding portion thereof, or both. It should be understood that "host cell" refers not only to a specific subject cell, but also to the offspring of such cells. Due to mutations or environmental influences, modifications may occur in offspring, so such offspring may actually be different from parental cells. However, such cells are still included within the scope of the term "host cell" as used herein.
[0529] Nucleic acid molecules encoding monoclonal antibodies or antigen-binding fragments thereof that specifically bind to BCMA according to the present invention and vectors containing these nucleic acid molecules can be used to transfect mammalian cells, plant cells, bacterial cells or yeast cells. Transfection can be carried out by any known method for introducing polynucleotides into host cells. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art, including dextran-mediated transfection, cationic polymer-nucleic acid complex transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, encapsulation of polynucleotides in liposomes, and direct microinjection of DNA into the nucleus. In addition, nucleic acid molecules can be introduced into mammalian cells by viral (expression) vectors.
[0530] Mammalian cell lines used as transformation hosts are well known in the art and include a variety of available immortalized cell lines. These include, for example, Chinese hamster ovary (CHO) cells, NS0 cells, SP2 cells, HEK-293T cells, FreeStyle293 cells (Invitrogen), NIH-3T3 cells, HeLa cells, baby hamster kidney (BHK) cells, African green monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549, SK-HEP1, HUH7, Hep-RG cells, and many other cell lines. Cell lines are selected by determining which cell lines have high expression levels and provide the necessary characteristics of the protein produced. Other cell lines that can be used are insect cell lines, such as Sf9 or Sf21 cells. When a recombinant expression vector encoding a monoclonal antibody or antigen-binding fragment thereof that specifically binds to BCMA is introduced into a mammalian host cell, the antibody or its fragment is produced as follows: the host cell is cultured for a period of time sufficient to allow the antibody or its fragment to be expressed in the host cell, or more preferably, the antibody or its fragment is secreted into the culture medium where the host cell grows. Monoclonal antibodies or antigen-binding fragments thereof that specifically bind to BCMA can be isolated from the culture medium using standard protein purification techniques. Plant host cells include, for example, Nicotiana, Arabidopsis, duckweed, corn, wheat, potato, etc. Bacterial host cells include Escherichia and Streptomyces species. Yeast host cells include Schizosaccharomyces pombe, Saccharomyces cerevisiae, and Pichia pastoris.
[0531] In addition, many known techniques can be used to enhance the production level of monoclonal antibodies or antigen-binding fragments thereof that specifically bind to BCMA from production cell lines. For example, the glutamine synthetase gene expression system (GS system) is a commonly used method to enhance expression under certain conditions.
[0532] Monoclonal antibodies or antigen-binding fragments thereof that specifically bind to BCMA from different cell lines may have different glycosylation profiles from each other. However, the monoclonal antibodies or antigen-binding fragments thereof that specifically bind to BCMA encoded by the nucleic acid molecules described herein or comprising the amino acid sequences provided herein are part of the present invention regardless of the glycosylation of the binding molecules and, in general, regardless of the presence or absence of post-translational modifications.
[0533] The above-mentioned host cells do not involve host cells produced using human embryos.
[0534] The above host cells do not relate to host cells produced by modifying the genetic integrity of human germline cells.
[0535] Methods of producing antibodies
[0536] In one aspect, the present invention relates to a method for producing an antibody or an antigen-binding fragment thereof that specifically binds to BCMA, comprising culturing the above-mentioned host cell in a growth medium under conditions sufficient to produce the antibody or fragment thereof, and then isolating and purifying the resulting antibody or fragment thereof, if necessary.
[0537] Pharmaceutical composition
[0538] Another aspect of the present invention is a pharmaceutical composition comprising as an active ingredient (or as the only active ingredient) a monoclonal antibody or an antigen-binding fragment thereof according to the present invention that specifically binds to BCMA.
[0539] In one aspect, the present invention relates to a pharmaceutical composition for treating a BCMA-mediated disease or condition, comprising a therapeutically effective amount of any of the above antibodies or antigen-binding fragments thereof and one or more pharmaceutically acceptable excipients.
[0540] "Pharmaceutical composition" means a composition comprising an antibody according to the present invention and at least one component selected from pharmaceutically acceptable and pharmacologically compatible fillers, solvents, diluents, carriers, adjuvants, dispensing agents and sensing agents, delivery agents.
[0541] The term "pharmaceutically acceptable" refers to one or more compatible liquid or solid components that are suitable for administration in mammals, preferably humans.
[0542] The term "excipient" is used herein to describe any ingredient other than the antibody according to the invention. These are substances of inorganic or organic nature used in the production / manufacturing of a drug in order to impart the necessary physicochemical properties to the drug.
[0543] In some embodiments, the composition is intended to ameliorate, prevent, or treat a disease or condition that may be mediated by BCMA.
[0544] The term "BCMA-mediated disease or disorder" refers to any disease or disorder that is directly or indirectly related to BCMA, including the etiology, development, progression, persistence, or pathology of the disease or disorder.
[0545] "Treatment," "treatment," and "therapy" refer to a method of alleviating or eliminating a biological disorder and / or at least one of its associated symptoms. Additionally, references herein to "treatment" include references to curative, palliative, and preventive treatments.
[0546] The term "disorder" refers to any condition that would benefit from treatment according to the present invention. The definition of this term includes chronic and acute disorders or diseases, including those pathological conditions that predispose a mammal to the disorder in question.
[0547] A "therapeutically effective amount" refers to the amount of therapeutic agent administered during the treatment period that will relieve to some extent one or more symptoms of the disease being treated. The therapeutically effective amount may vary depending on a variety of factors, such as the specific condition being treated, the age, sex, and weight of the patient, and whether the monoclonal antibody or antigen-binding fragment thereof that specifically binds to BCMA is administered as a stand-alone monotherapy or in combination with one or more additional drugs or therapies.
[0548] In one aspect, the subject or patient treated is a mammal, preferably a human subject. The subject can be male or female of any age.
[0549] The pharmaceutical composition of the present invention and its preparation method are undoubtedly obvious to those skilled in the art. The pharmaceutical composition should preferably be manufactured according to GMP (Good Manufacturing Practice) requirements.
[0550] In some embodiments of the pharmaceutical composition, it may include a buffer composition, a tonicity agent (osmotic agent or osmotic agent), a stabilizer and / or a solubilizing agent.
[0551] The pharmaceutical composition according to the present invention is a stable composition.
[0552] A pharmaceutical composition is "stable" if the active agent maintains its physical stability and / or chemical stability and / or biological activity over a specified shelf life at storage temperature (e.g., 2-8°C). Preferably, the active agent retains physical and chemical stability as well as biological activity. The shelf life is adjusted based on the results of stability testing under accelerated or natural aging conditions.
[0553] The pharmaceutical composition according to the present invention is suitable for parenteral administration as a sterile formulation, and is intended to be administered in a subject by bypassing the gastrointestinal tract through a rupture of the skin or mucosal barrier, by injection, infusion and implantation. In particular, parenteral administration is contemplated, including, inter alia, subcutaneous, intraperitoneal, intramuscular, intravenous, intraarterial, intrathecal, intraventricular, intraurethral, intracranial, intrasynovial, transdermal injection or infusion; and renal dialysis infusion techniques. Intratumoral delivery, such as intratumoral injection, may also be employed. Regional perfusion is also contemplated.
[0554] In some embodiments, the pharmaceutical composition is administered intravenously.
[0555] In some embodiments, intravenous administration is performed by using infusion, extended infusion, or continuous infusion over time.
[0556] In some embodiments, the pharmaceutical composition is administered subcutaneously.
[0557] In some embodiments, subcutaneous administration is performed by using subcutaneous injection.
[0558] In some embodiments, the pharmaceutical composition is an injectable dosage form.
[0559] In some embodiments, the injectable dosage form is an infusion solution.
[0560] In some embodiments, the injectable dosage form is a solution for subcutaneous administration.
[0561] Injectable preparations can be manufactured without limitation in unit dosage forms, such as ampoules, vials, plastic containers, prefilled syringes, automatic injection devices.
[0562] In some embodiments, the pharmaceutical composition is a pharmaceutical composition provided in dry form, i.e., in powder or granular form, for reconstitution with a suitable solvent (e.g., sterile pyrogen-free water) prior to administration. Such pharmaceutical preparations can be prepared, for example, by lyophilization, a freeze-drying process known in the art, and which involves freezing the product and then removing the solvent from the frozen material.
[0563] In some embodiments, the pharmaceutical composition is a lyophilisate for preparing an infusion solution.
[0564] In some embodiments, the pharmaceutical composition is a lyophilisate for preparing a solution for subcutaneous administration.
[0565] In some embodiments, the pharmaceutical composition is a concentrate for preparing an infusion solution.
[0566] In some embodiments, the pharmaceutical composition is a concentrate for preparing a solution for subcutaneous administration.
[0567] In one aspect, the present invention relates to a pharmaceutical composition comprising a monoclonal antibody or an antigen-binding fragment thereof according to the present invention that specifically binds to BCMA and at least one other therapeutically active compound.
[0568] In one aspect, the invention relates to a pharmaceutical composition for treating a BCMA-mediated disease or condition, comprising any of the above antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound.
[0569] In one aspect, the invention relates to a pharmaceutical composition comprising any of the above-mentioned antibodies or antigen-binding fragments thereof and further at least one other therapeutically active compound.
[0570] In one aspect, the present invention relates to a pharmaceutical composition for treating a BCMA-mediated disease or condition, comprising any of the above antibodies or antigen-binding fragments thereof and further at least one other therapeutically active compound.
[0571] In one aspect, the present invention relates to a pharmaceutical composition for treating a BCMA-mediated disease or condition, comprising any of the above antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound, which is an antibody, a small molecule, a hormonal therapeutic agent, or a combination thereof.
[0572] In some embodiments of the pharmaceutical composition, the BCMA-mediated disease or condition is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
[0573] Therapeutic use of monoclonal antibodies or antigen-binding fragments thereof that specifically bind to BCMA
[0574] In one aspect, an antibody or antigen-binding fragment thereof that specifically binds BCMA is used to treat a disorder mediated by BCMA activity.
[0575] In one aspect, the subject or patient treated is a mammal, preferably a human subject. The subject can be male or female of any age.
[0576] In one aspect, the invention relates to a method for treating a BCMA-mediated disease or disorder, comprising administering any of the above-described antibodies or antigen-binding fragments thereof or the pharmaceutical composition in a therapeutically effective amount to a subject in need of such treatment.
[0577] In one aspect, the invention relates to a method for treating a BCMA-mediated disease or condition comprising administering to a subject in need of such treatment any of the above antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound in a therapeutically effective amount.
[0578] In some embodiments of the methods of treatment, the BCMA-mediated disease or disorder is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
[0579] In some embodiments of the methods of treatment, the other therapeutically active compound is a small molecule, a hormonal therapeutic agent, or any combination thereof.
[0580] In one aspect, the present invention relates to the use of the above-mentioned antibody or antigen-binding fragment thereof or the above-mentioned pharmaceutical composition for treating a BCMA-mediated disease or disorder in a subject in need of such treatment.
[0581] In one aspect, the present invention relates to the use of the above-mentioned antibodies or antigen-binding fragments thereof and at least one other therapeutically active compound for treating a BCMA-mediated disease or disorder.
[0582] In some embodiments of use, the BCMA-mediated disease or disorder is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
[0583] In some embodiments of the use, the other therapeutically active compound is a small molecule, a hormonal therapeutic agent, or any combination thereof.
[0584] As used herein, a use or method involving an antibody or antigen-binding fragment thereof that specifically binds BCMA and one or more other therapeutic agents is intended to mean, refer to, and include the following:
[0585] 1) administering such a combination of an antibody or antigen-binding fragment thereof that specifically binds BCMA and a therapeutic agent simultaneously to a patient in need of treatment, where such components are formulated together into a single dosage form that releases said components to said patient substantially simultaneously,
[0586] 2) administering such a combination of an antibody or antigen-binding fragment thereof that specifically binds to BCMA and a therapeutic agent simultaneously to a patient in need of treatment, wherein such components are formulated separately from each other into separate dosage forms that are taken by the patient at substantially the same time, whereby the components are released to the patient at substantially the same time,
[0587] 3) sequentially administering such a combination of an antibody or antigen-binding fragment thereof that specifically binds BCMA and a therapeutic agent to a patient in need of treatment, wherein such components are formulated separately from one another into separate dosage forms that are taken by the patient at consecutive times with distinct time intervals between each administration, whereby the components are released to the patient at distinctly different times; and
[0588] 4) sequential administration of such a combination of an antibody or antigen-binding fragment thereof that specifically binds to BCMA and a therapeutic agent to a patient in need of treatment, where such components are formulated together into a single dosage form that releases the components in a controlled manner, whereby they are released to the patient simultaneously, sequentially or co-integrated at the same and / or different times, wherein each part can be administered by the same or different routes.
[0589] The antibody or antigen-binding fragment thereof that specifically binds BCMA can be administered in the absence of other therapeutic treatments, ie, as a stand-alone therapy.
[0590] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with a proteasome inhibitor.
[0591] In some embodiments of the treatment methods or uses, an antibody or antigen-binding fragment that specifically binds BCMA can be administered in combination with an anti-tumor immunomodulator (e.g., lenalidomide, pomalidomide).
[0592] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA can be administered in combination with cytostatic chemotherapy (cyclophosphamide, etoposide, etc.).
[0593] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with a tyrosine kinase inhibitor.
[0594] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with a BCL2 inhibitor product.
[0595] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with an exportin 1 antagonist product.
[0596] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with a glucocorticoid.
[0597] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA can be administered in combination with an anti-tumor monoclonal antibody (e.g., daratumumab).
[0598] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with a targeted therapy.
[0599] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA can be administered in combination with a monoclonal antibody that agonizes cytokines (eg, IL15SA, IL2).
[0600] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA can be administered in combination with a monoclonal antibody that antagonizes cytokines (eg, anti-IL6R, anti-TNF).
[0601] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with a G-CSF (granulocyte colony-stimulating factor) product.
[0602] In some embodiments of the method of treatment or use, an antibody or antigen-binding fragment that specifically binds BCMA can be co-administered with hematopoietic stem cell transplantation.
[0603] In some embodiments of the methods of treatment or uses, an antibody or antigen-binding fragment that specifically binds BCMA may be administered in combination with radiation therapy.
[0604] In some embodiments of the methods of treatment or use, a suitable dosage of a monoclonal antibody or antigen-binding fragment thereof that specifically binds to BCMA according to the present invention will be in the range of 0.1 to 200 mg / kg. BRIEF DESCRIPTION OF THE DRAWINGS
[0605] Figure 1 is a map of an expression vector carrying the genetic sequence of the variable domains.
[0606] Figure 2 is a map of an expression vector carrying the genetic sequence of the variable domains.
[0607] for Figures 1 to 2
[0608]
[0609]
[0610] Figure 3 It is a vector map with the genetic sequence of the antibody heavy chain.
[0611] Figure 4 It is a vector map with the genetic sequence of the antibody light chain.
[0612] for Figures 3 to 4
[0613]
[0614]
[0615] Figure 5 The electrophoresis of candidates 01, 06, 016, 021 and 022 in 7.5% polyacrylamide gel under denaturing non-reducing conditions after the first stage purification on protein A adsorbent.
[0616] M is a molecular weight marker.
[0617] 1-Candidate 01.
[0618] 2-Candidate 06.
[0619] 3-Candidate 016.
[0620] 4-Candidate 021.
[0621] 5-Candidate 022.
[0622] Figure 6is the electrophoresis of candidates 024, 025, 026, 027, 030, 033 and 037 in 7.5% polyacrylamide gel under denaturing non-reducing conditions after the first stage purification on protein A adsorbent.
[0623] M is a molecular weight marker.
[0624] 1-Candidate 024.
[0625] 2-Candidate 025.
[0626] 3-Candidate 026.
[0627] 4-Candidate 027.
[0628] 5-Candidate 030.
[0629] 6-Candidate 033.
[0630] 7-Candidate 037. Example
[0631] The following examples are provided for a better understanding of the present invention. These examples are for illustrative purposes only and should not be construed as limiting the scope of the present invention in any way.
[0632] All publications, patents and patent applications cited in this specification are incorporated herein by reference. Although the foregoing invention has been described in some detail by way of illustration and example for the purpose of clear understanding, it will be apparent to those skilled in the art that certain changes and modifications may be made based on the teachings of the present invention without departing from the spirit or scope of the attached embodiments.
[0633] Materials and general methods
[0634] For general information about the nucleotide sequences of human immunoglobulin light and heavy chains, see: Kabat, EA, et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991). The amino acids of the antibody chains are numbered according to the EU numbering (Edelman, GM, et al., Proc. Natl. Acad. Sci. USA 63 (1969) 78-85; Kabat, EA, et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991).
[0635] Recombinant DNA technology
[0636] DNA was manipulated using standard methods, such as described in Sambrook, J. et al, Molecular cloning: A laboratory manual; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989. Molecular biology reagents were used according to the manufacturers' protocols.
[0637] Gene synthesis
[0638] The desired gene segments were prepared from oligonucleotides generated by chemical synthesis. Gene segments 300-1400 bp long flanked by single restriction sites were assembled by annealing and ligation of oligonucleotides (including PCR amplification) and then cloned through restriction sites. The DNA sequence of the subcloned gene fragments was confirmed by DNA sequencing.
[0639] DNA sequencing
[0640] DNA sequences were determined by Sanger sequencing.
[0641] DNA and protein sequence analysis and sequence data management
[0642] Unipro's UGENE suite version 1.29 and SnapGene Viewer were used for sequence creation, plotting, analysis, annotation, and interpretation.
[0643] Expression vector
[0644] In order to express the antibodies described in the present application materials, variants of expression plasmids intended for expression in prokaryotic cells (E. coli), transient expression in eukaryotic cells (e.g., in CHO cells) were used. In addition to the antibody expression cassette, the vector also contains: an origin of replication that allows the plasmid to replicate in E. coli, genes that confer resistance to various antibiotics (e.g., ampicillin, kanamycin) on E. coli.
[0645] The fusion gene comprising the antibody chain described below is produced by PCR and / or gene synthesis, and is assembled by connecting the corresponding nucleic acid segments (e.g., using the unique restriction sites in the corresponding vector) using known recombination methods and techniques. The nucleic acid sequence of the subclone is verified by DNA sequencing. For transient transfection, a larger amount of plasmid is prepared by preparing plasmids from transformed E. coli cultures.
[0646] Example 1. Production of recombinant antigens in mammalian cell suspension cultures
[0647] The extracellular domain sequence of human BCMA receptor (SEQ https: / / www.uniprot.org / uniprot / Q02223 (1-54aa)) was cloned into pEE plasmid for protein production in mammalian cells using TEV-Fc, Avi-His, Avi-His-TEV-HSA tags through SalI / NotI restriction sites. The required amount of plasmid was grown in E. coli cells and purified using Qiagen kit.
[0648] Antigens are produced in an established cell line derived from Chinese Hamster Ovary (CHO-T) cells. Suspension cultures are performed in flasks on an orbital incubator shaker. To express the target antigen, cells are transfected using linear polyethyleneimine. Nine days after transfection, the culture fluid is separated from the cells by filtration through a 0.5 / 0.22 μm deep bed filter.
[0649] The antigen BCMA-TEV-Fc was isolated from the culture fluid and purified using a column with a protein A affinity chromatography adsorbent. The clarified culture fluid was passed through a column balanced with phosphate buffered saline (PBS, pH 7.4). The bound antigen was eluted using 0.1M glycine buffer (pH 2.5). The resulting target protein was then dialyzed into PBS (pH 7.4), DTT was added, the mixture was filtered through 0.22μm Millex GP, transferred to a test tube and stored at -70°С. BCMA-Avi-His and BCMA-Avi-His-TEV-HSA antigens were isolated from the culture fluid and purified using a Ni-NTA affinity chromatography column. The clarified culture fluid was passed through a column balanced with phosphate buffered saline (PBS, pH 7.4). The bound antigen was eluted with PBS supplemented with 500mM imidazole (pH7.4). The protein was then dialyzed into PBS. Thereafter, the protein was applied to a gel filtration column. The resulting protein was filtered through 0.22 μm Millex GP, transferred into tubes and stored at -70°C.
[0650] Example 2. Cloning of variable domain genes of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 into expression plasmids
[0651] The variable domain genes of the antibodies were cloned into the expression plasmid pLL ( Figure 1 ) and pET22( Figure 2 )middle.
[0652] Subsequently, the expression vectors containing the antibody fragments were transformed into E. coli BL21-DE3 and BL21-Gold strains to compare the affinity of the variable antibody fragments from the display library to the antigen by ELISA using an automated platform.
[0653] Example 3. Production and preliminary analysis of Fab and scFv of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 that specifically bind to human BCMA
[0654] Fab and scFv were generated according to standard techniques: E. coli BL21-DE3 and BL21-Gold bacterial cells were transformed with expression vectors containing scFv and Fab genes, respectively, and then the transcription of the lac operon was triggered by the addition of inducers, thereby culturing the resulting transformants simultaneously, resulting in the expression of scFv and Fab, and their export to the periplasmic space. ELISA was then performed to verify the binding of scFv and Fab to the substrate-immobilized antigen hBCMA-Avi-His / 0.1 M NaHCO at a concentration of 0.2 μg / ml. 3 (pH 9.0) (antigen was fixed overnight at 4°C). All further steps were performed at room temperature according to standard ELISA protocols using a high-throughput automated platform based on a robotic system. Washing with 300 μl / well 1x PBST was repeated 3 times after each step. Nonspecific binding sites on the plate were blocked with 1x PBST containing 1% skim milk; after washing, analyte (60 μl / well E. coli supernatant) was added. The immune complex was detected using a peroxidase-conjugated goat anti-Myc tag antibody (1:20,000). The substrate color development mixture was stained by adding 50 μl of substrate solution for 15 minutes. 25 μl of 1% H 2 SO 4 The reaction was stopped. The color signal was measured at 450 nm wavelength. The level of antibody binding was proportional to the signal generated. Clones with a color signal exceeding the control antibody signal were tested in an ELISA for nonspecific binding.
[0655] Thus, all tested Fabs and scFvs of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 showed specific binding to hBCMA-Avi-His.
[0656] Example 4. Analysis of nonspecific binding of Fab and scFv of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 to other antigens
[0657] ELISA was also used to analyze nonspecific binding of the scFv and Fab in question to different antigens. The assay was performed as described above, except that 0.1 M NaHCO 3hCD16-Avi-His-FLAG, cynoIL4R-Fc in (pH 9.0) were used as immobilized antigens (antigens were immobilized overnight at 4°C). hBCMA-Avi-His, rhesus BCMA were used as controls for specific binding (antigens were immobilized overnight at 4°C). All subsequent steps were performed according to standard ELISA protocols using a high-throughput automated platform.
[0658] All tested Fabs and scFvs of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 showed specific binding to hBCMA-Avi-His, rhesus BCMA, and did not show non-specific interactions with hCD16-Avi-His-FLAG and cynoIL4R-Fc.
[0659] Example 5. Analysis of the off-rates of Fab and scFv of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 on ForteBio
[0660] Fab was measured on a ForteBio Octet RED 384 system using a FAB2G biosensor (ForteBio). Fab samples were loaded onto the sensor from the supernatant at (5±3)°C for 16-18 hours. After loading, the sensor was transferred to a kinetic buffer solution containing 500 mM NaCl (4.3 mM Na 2 HPO 4 ; 500 mM NaCl, 1.5 mM KH 2 PO 4; 2.7mM KCl; the volume fraction of Tween20 added was 0.1%; the mass fraction of BSA added was 0.1%; pH7.4), the given buffer solution was used to prepare all subsequent experimental steps involving Fab samples. The measurements were performed at 30°C. The sensor was equilibrated in the kinetic buffer solution for at least 10 minutes, and then the baseline was recorded (60 seconds). After the baseline recording, the sensor with the immobilized Fab was immersed in a well containing the analyte solution (recombinant human antigen BCMA-Avi-His-TEV-HSA), where the antigen-Fab complex was bound for 300 seconds. The concentration of human BCMA was 391.2nM (30μg / ml). The dissociation of the complex in the kinetic buffer solution was then detected for 600 seconds. In order to check for nonspecific interactions between the analyte and the sensor (negative control), we used a sensor without the candidate Fab. In the loading step, the negative control sensor was immersed in the supernatant without Fab, and all other steps were similar to those used for the sensor loaded with the Fab sample.
[0661] scFv was measured on a Forte Bio Octert RED 384 system using a SAX biosensor (ForteBio). For the measurements, human BCMA was biotinylated at an antigen / biotin molar ratio of 1:1.5. Interaction kinetics analysis was performed at 30°C. Kinetic buffer (4.3 mM Na 2 HPO 4 ; 136.9 mM NaCl, 1.5 mM KH 2 PO 4 ; 2.7mM KCl; the volume fraction of Tween20 added was 0.1%; the mass fraction of BSA added was 0.1%; pH 7.4) for analysis. Biotinylated human BCMA was loaded onto the sensor for 600 seconds, and the concentration of the antigen solution in the kinetic buffer solution was 20μg / ml. The baseline was recorded for 120 seconds. After the baseline was recorded, the sensor with the fixed antigen was immersed in a well containing the analyte solution (candidate scFv), where the antigen-scFv complex was bound for 150 seconds (for the binding step, the scFv sample was diluted with the kinetic buffer solution to a concentration of 1200nM). The dissociation of the antigen-scFv complex in the kinetic buffer solution was then detected for 600 seconds. The reference signal was recorded simultaneously with the recording of the candidate sensorgram and was subtracted when the sensorgram was processed. The reference signal is the signal of the sensor immersed in the kinetic buffer solution without analyte during the binding step, and all other steps are similar to the sensor recording the test signal. The sensor without antigen loading is used to verify the nonspecific interaction between the analyte (candidate scFv) and the sensor. In the loading step, the negative control sensor was immersed in the sodium kinetic buffer solution without antigen, and all other steps were similar to those for the antigen-loaded sensor.
[0662] Octet Data Analysis (version 9.0) software was used to analyze the binding curves using a 1:1 interaction model. The results of the analysis of Fab and scFv of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 are shown in Table 1. These Fab and scFv were further used to generate full-length antibodies.
[0663] Table 1 - Kinetic analysis of the interaction of Fab and scFv of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 with human BCMA on ForteBio Octet RED384.
[0664]
[0665] Example 6. Generation of genetic constructs for synthesizing full-length antibodies
[0666] Cloning was performed by standard techniques. We generated PCR products containing the heavy and light chain variable domain genes of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 using primers containing restriction sites. The heavy chain variable domain was cloned into the vector pEE-HCLALA IgG1 ( Figure 3 The light chain variable domain was cloned into the pEE-CK vector via the Sal1 / BsiW1 restriction sites ( Figure 4 ). The resulting gene construct was transferred for transient protein production in a CHO-T cell line.
[0667] Example 7. Production of full-length antibodies
[0668] Full-length antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 were produced in duplicate using transient transfection in an established cell line derived from Chinese hamster ovary cells (CHO-T). Suspension cultures were performed in an orbital shaking bioreactor using serum-free medium. For transient expression, linear polyethyleneimine was used at 2-2.2×106 Cells were transfected at a concentration of 10 cells / ml. The DNA / PEI ratio was 1:7. On the 10th day of culture, the cell suspension was centrifuged at 2000 g for 15 minutes and filtered through a 0.22 μm filter.
[0669] The antibody was purified by affinity chromatography using a robotic workstation. The column was balanced with a buffer containing 50mM NaPB (sodium phosphate buffer), 150mM NaCl (pH 7.5), the filtered culture solution containing the antibody was applied, and then the column was washed with 8 volumes of a buffer containing 50mM NaPB, 150mM NaCl and 4 volumes of 50mM NaPB (pH 7.5). The protein was eluted using 6 column volumes of a 50mM NaPB, 100mM NaCl (pH 3) solution. The resulting sample was neutralized with 25μl of 1M phosphate buffer (pH 8).
[0670] The purity of the candidate was monitored using polyacrylamide gel electrophoresis and size exclusion HPLC. Electrophoresis was performed in 7.5% polyacrylamide gel under denaturing non-reducing conditions. Protein purity was determined by the intensity of band staining at 10 μg protein load per lane. Figure 5 and Figure 6 Size exclusion HPLC was performed on the column with a mobile phase of 0.05 M NaH 2 PO 4 , 0.3M NaCl pH=7.0.
[0671] Example 8. Determination of the affinity of full-length antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 to human BCMA on Forte Bio
[0672] The experiments investigating the affinity of antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 to human BCMA were performed on a ForteBio Octet RED 384 system. Antibodies at a concentration of 20 μg / ml were immobilized on the surface of a protein A biosensor (ForteBio). The analysis was performed at 30°C using a kinetic buffer solution (4.3 mM Na 2 HPO 4 ; 136.9 mM NaCl, 1.5 mM KH 2 PO 4; 2.7mM KCl; the volume fraction of Tween 20 added was 0.1%; the mass fraction of BSA added was 0.1%; pH 7.4). After setting the baseline in the kinetic buffer solution, the sensor with the immobilized antibody was immersed in a well containing the analyte (BCMA) solution, in which the complex bound for 300 seconds. For each test antibody, the sensorgram of the human BCMA solution with a concentration of 10μg / ml (130.4nM) and the reference signal (reference sensorgram) of the kinetic buffer solution without BCMA were recorded. The dissociation of the complex in the buffer solution was then detected for 600 seconds. In order to verify the nonspecific interaction between the analyte and the sensor (negative control), we used a sensor without antibody loading. In the loading step, the negative control sensor was immersed in a sodium acetate buffer without antibody, and all other steps were similar to those used for the sensor loaded with the antibody.
[0673] After subtracting the reference signal, binding curves were analyzed using Octet Data Analysis (version 9.0) software and using a 1:1 interaction model (Table 2).
[0674] Table 2 - Kinetic constants for antibody / human BCMA interactions.
[0675]
[0676]
[0677] Thus, all tested anti-BCMA antibodies specifically bound to human BCMA and had high binding affinity parameters relative to human BCMA (Table 3).
[0678] Example 13. Validation of the interaction between full-length antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 and rhesus monkey BCMA on the Forte Bio system.
[0679] This experiment was designed to confirm the interaction between antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 and rhesus BCMA. The interaction between the antibodies and rhesus BCMA was validated on the ForteBio Octet RED384 system using AR2G biosensors (ForteBio). The experiment included the following steps: activation of the sensor, loading of proteins onto the sensor, quenching of unreacted activation groups, recording baseline, recording analyte binding, and recording dissociation. The measurements were performed at 30°C. The sensor was activated for 300 seconds in an aqueous solution containing 20 mM EDC and 10 mM sNHS. Rhesus BCMA was loaded onto the biosensor surface in a 10 mM sodium acetate buffer solution at pH 5.0 for 300 seconds. The concentration of the loaded protein was 10 μg / ml. The unreacted active sites on the sensor surface were quenched in a 1 M ethanolamine aqueous solution at pH 8.5 for 300 s (pH was adjusted by adding hydrochloric acid). To verify the nonspecific interaction between the analyte and the sensor (negative control), we used an antigen-free sensor (during the loading step, the sensor was immersed in a 10 mM sodium acetate buffer solution at pH 5.0; all other steps were similar to those for the antigen-loaded sensor). The baseline and all subsequent steps of the experiment were performed in a kinetic buffer solution (4.3 mM Na 2 HPO 4 ; 136.9mM NaCl, 1.5mM KH 2 PO 4 ; 2.7mM KCl; 0.1% Tween 20 added by volume; 0.1% BSA added by mass; pH 7.4). In the binding step (step duration of 300 seconds), the rhesus monkey BCMA loaded sensor was immersed in the well containing the analyte (test antibody) solution prepared in the kinetic buffer solution. The analyte concentration was 2.5μg / ml. In the dissociation step, the sensor was immersed in the well containing the kinetic buffer, where the baseline was recorded.
[0680] Octet Data Analysis (version 9.0) software was used to process the binding curves using a 1:1 interaction model. The results of the processing are shown in Table 4 (kdis values <1.0E-07 (1 / s) and corresponding KD values <1.0E-12 (M) mean that, within the framework of the applied measurement and processing techniques, the given sample shows a sensitivity limit of the dissociation rate constant that exceeds the model curve describing the sensorgram). By using a bivalent analyte (antibody), the resulting kinetic constant values are of an estimated nature and can only be used to compare samples. All anti-BCMA antibodies showed a signal for binding to rhesus BCMA in the binding step and a slow signal decay in the dissociation step, confirming the high affinity when interacting with the antigen. Therefore, all tested anti-BCMA antibodies specifically bind to rhesus BCMA (Table 3).
[0681] Table 3 - Processing results of antibody-rhesus BCMA binding sensorgrams.
[0682]
[0683] Example 14. Validation of the inhibition of the interaction between APRIL and human BCMA by antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 on the Forte Bio system.
[0684] The study was performed on a ForteBio Octet RED 384 system using an AR2G biosensor (ForteBio). The experiment included the following main steps: Human APRIL / TNFSF13 antigen was loaded onto the sensor and binding of APRIL / TNFSF13 antigen to a mixture of human BCMA and test antibodies (premix) was verified. A human BCMA solution without antibody was used as a positive control. Inhibition of the interaction was determined by comparing the premix signal (response) with the positive control signal.
[0685] The sensor was activated in an aqueous solution containing 20 mM EDC and 10 mM sNHS for 300 s. APRIL / TNFSF13 antigen was loaded onto the biosensor surface in 10 mM sodium acetate buffer, pH 5.0, for 900 s. The APRIL / TNFSF13 protein loading concentration was 10 μg / ml. To verify the nonspecific interaction between the analyte and the sensor, we used an antigen-free sensor (during the loading step, the sensor was immersed in sodium acetate buffer, pH 5.0; all other steps were similar to those used for the antigen-loaded sensor). The unreacted active centers on the sensor surface were quenched in a 1 M ethanolamine aqueous solution at pH 8.5 (pH adjusted by adding hydrochloric acid) for 300 s. All experimental steps after the quenching step were performed in kinetic buffer: (4.3 mM Na 2 HPO 4 ; 136.9 mM NaCl, 1.5 mM KH 2 PO 4 ; 2.7 mM KCl; the volume fraction of Tween 20 added was 0.1%; the mass fraction of BSA (bovine serum albumin) added was 0.1%; pH 7.4). In the binding step (step duration of 300 seconds), the protein-loaded sensor was immersed in the well containing the analyte solution. A solution containing 710 nM test antibody and 71 nM human BCMA in a kinetic buffer solution was used as the analyte in the binding step. A solution containing human BCMA at a concentration of 71 nM (analyte) was used as a positive control. The measurement was performed at 30°C. The reference sensor underwent all the same steps as the sensor used to record the analyte sensorgram, except for the binding step - in the binding step, the sensor was immersed in a kinetic buffer without analyte (the reference sensor signal was measured in parallel and the main sensorgram was recorded). When processing the sensorgram, the reference signal was subtracted from the analytical signal.
[0686] The sensorgrams were processed using ForteBio Octet Data Analysisn 9.0 software. The experimental results are shown in Table 4. Test antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 inhibited the binding of APRIL / TNFSF13 loaded on the sensor to human BCMA.
[0687] Table 4 - Validation results of antibodies inhibiting the interaction between APRIL / TNFSF13 and human BCMA.
[0688]
[0689]
[0690] Example 15. Analysis of the binding of anti-BCMA antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 001-033, 01-037 to human BCMA on the surface of RPMI8226 cells by flow cytometry.
[0691] The ability of anti-BCMA antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 to bind to human BCMA on the surface of RPMI8226 cell line (multiple myeloma) that stably expresses the BCMA receptor on its surface was evaluated by flow cytometry.
[0692] 25,000 cells in a well were plated in staining buffer (PBS + NaN 3 +5% BSA) incubated at +4°C for 30 minutes, and the test antibody was serially diluted. Thereafter, the cells were washed twice with cold (+4°C) staining buffer, and the bound antibodies were detected by staining with phycoerythrin-labeled anti-human Fc secondary antibodies at +4°C for 30 minutes. Wells in which no antibody was introduced and only with secondary antibodies were used as controls. Thereafter, the cells were washed twice with cold (+4°C) staining buffer and resuspended in 150 μl of cold (+4°C) staining buffer. The samples were then analyzed on a flow cytometer. We performed forward and side scatter (FSC and SSC) gating to distinguish the target cell population; thereafter, we performed forward scatter height and forward scatter area (FSC-H and FSC-A) gating to identify individual (single) cells in the population.
[0693] Calculate EC by drawing a four-parameter logistic regression model 50 The half effective concentration is shown in Table 5.
[0694] Table 5 – ED of anti-BCMA antibodies when bound to the surface of RPMI8226 cells 50 .
[0695] Antibody <![CDATA[EC 50 (μg / ml)]]> 01-001 0.59 01-006 0.45 01-016 1.05 01-021 0.49 01-022 0.76 01-024 0.22 01-025 0.70 01-026 1.75 01-027 0.25 01-030 0.33 01-033 0.39 01-037 0.79
[0696] Conclusions: The data obtained show that all antibodies tested bind to the BCMA target on the surface of multiple myeloma cells.
[0697] Example 16. Analysis of non-specific binding of anti-BCMA antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 on the surface of BCMA-negative K562 cells by flow cytometry.
[0698] The presence of non-specific binding of anti-BCMA antibodies 01-001, 01-006, 01-016, 01-021, 01-022, 01-024, 01-025, 01-026, 01-027, 01-030, 01-033, 01-037 was evaluated by flow cytometry on the K562 cell line (chronic myeloid leukemia), which is characterized by the absence of BCMA expression on its surface.
[0699] 25,000 K562 cells in a well were incubated with diluted test antibodies in staining buffer (PBS + NaN 3 +5% BSA) at +4°C for 30 minutes. Thereafter, the cells were washed twice with cold (+4°C) staining buffer and the bound antibodies were detected by staining with phycoerythrin-labeled anti-human Fc secondary antibodies at +4°C for 30 minutes. Wells in which no antibody was introduced and only with secondary antibodies were used as controls. Thereafter, the cells were washed twice with cold (+4°C) staining buffer and resuspended in 150 μl of cold (+4°C) staining buffer. The samples were then analyzed on a flow cytometer. We performed forward and side scatter (FSC and SSC) gating to distinguish the target cell population; thereafter, we performed forward scatter height and forward scatter area (FSC-H and FSC-A) gating to identify individual (single) cells in the population.
[0700] RPMI8226 was used as a positive control to test the cell surface binding of antibodies to BCMA. Sample preparation was performed similarly to K562 described above.
[0701] The non-specific binding of the antibodies was assessed by comparing the fluorescence signal levels of the same concentrations of antibodies in K562 and RPMI8226 cell line variants, and the data are shown in Table 6.
[0702] Table 6 - Cell surface binding data of anti-BCMA antibodies to K562 (BCMA negative line) and RPMI8226 (BCMA positive line).
[0703]
[0704]
[0705] Conclusion: The results showed that all antibodies tested showed no nonspecific binding to BCMA-negative cell lines.
Claims
1. An isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to BCMA, comprising: (a) a heavy chain variable domain comprising: (i) a CDR1 having the amino acid sequence SX1X2MS, wherein X1=S or G; X2=A, L or V; (ii) a CDR2 having the amino acid sequence X3YNGGSDRAGX4X5DSVX6G, wherein X3=G or C; X4=F or Y; X5=A or T; X6 = E or K; and (iii) a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: (i) having the amino acid sequence X7GX8X9SNIGX 10 X 11 X 12 X 13 VX 14 CDR1, in X7 = S or T; X8 = S, G or T; X9=S, T, I or R; X 10 =0 or A; X 11 =S, H, N, G or T; X 12 =N, R or Y; X 13 =T, A, I or D; X 14 =N or H; (ii) having the amino acid sequence X 15 X 16 X 17 X 18 CDR2 of RPS, X15 = N, R, K, S or G; X16 = D, N, G, T or H; X17 = S, N or T; X18 = Q or N; and (iii) having the amino acid sequence X 19 X 20 WDX 21 X 22 X 23 X 24 X 25 WX 26 CDR3, in X 19 =A or S; X 20 =A, S, T or V; X 21 =G, D, H or S; X 22 =S, D or R; X 23 =L or V; X 24 =N, T, R or S; X 25 =V, A, G or N; X 26 =M, V or L.
2. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO:
3.
3. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable domain comprises a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO:
6.
4. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the light chain variable domain comprises a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO:
15.
5. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the light chain variable domain comprises a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, or SEQ ID NO:
24.
6. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the light chain variable domain comprises a CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, or SEQ ID NO:
35.
7. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises: A CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6; and A CDR3 having the amino acid sequence of SEQ ID NO:
7.
8. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 7, comprising: (i) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; (ii) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 5 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; (iii) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; (iv) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4 and A CDR3 having the amino acid sequence of SEQ ID NO: 7; or (v) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 6 and A CDR3 having the amino acid sequence of SEQ ID NO:
7.
9. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, comprising a light chain variable domain, wherein the light chain variable domain comprises: a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15; a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24; and A CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34 or SEQ ID NO:
35.
10. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 9, comprising: (i) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 8, A CDR2 having the amino acid sequence of SEQ ID NO: 16 and a CDR3 having the amino acid sequence of SEQ ID NO: 25; (ii) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO:9, A CDR2 having the amino acid sequence of SEQ ID NO: 17 and a CDR3 having the amino acid sequence of SEQ ID NO: 26; (iii) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 10, A CDR2 having the amino acid sequence of SEQ ID NO: 18 and a CDR3 having the amino acid sequence of SEQ ID NO: 27; (iv) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 11, A CDR2 having the amino acid sequence of SEQ ID NO: 19 and a CDR3 having the amino acid sequence of SEQ ID NO: 28; (v) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 12, A CDR2 having the amino acid sequence of SEQ ID NO: 20 and a CDR39 having the amino acid sequence of SEQ ID NO: 29; (vi) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 12, A CDR2 having the amino acid sequence of SEQ ID NO: 20 and a CDR3 having the amino acid sequence of SEQ ID NO: 30; (vii) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 10, A CDR2 having the amino acid sequence of SEQ ID NO: 21 and a CDR3 having the amino acid sequence of SEQ ID NO: 31; (viii) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 13, A CDR2 having the amino acid sequence of SEQ ID NO: 22 and a CDR3 having the amino acid sequence of SEQ ID NO: 32; (ix) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 14, A CDR2 having the amino acid sequence of SEQ ID NO: 23 and a CDR3 having the amino acid sequence of SEQ ID NO: 33; (x) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 8, A CDR2 having the amino acid sequence of SEQ ID NO: 24 and A CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xi) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 15, A CDR2 having the amino acid sequence of SEQ ID NO: 17 and A CDR3 having the amino acid sequence of SEQ ID NO:
35.
11. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, comprising: (a) a heavy chain variable domain comprising: A CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3; a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6; and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: a CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15; a CDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23 or SEQ ID NO: 24; and A CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34 or SEQ ID NO:
35.
12. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 11, comprising: (i)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 1, CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 8, A CDR2 having the amino acid sequence of SEQ ID NO: 16 and a CDR3 having the amino acid sequence of SEQ ID NO: 25; (ii)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 5 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO:9, A CDR2 having the amino acid sequence of SEQ ID NO: 17 and a CDR3 having the amino acid sequence of SEQ ID NO: 26; (iii)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 10, A CDR2 having the amino acid sequence of SEQ ID NO: 18 and a CDR3 having the amino acid sequence of SEQ ID NO: 27; (iv)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 11, A CDR2 having the amino acid sequence of SEQ ID NO: 19 and a CDR3 having the amino acid sequence of SEQ ID NO: 28; (v)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 5 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 12, A CDR2 having the amino acid sequence of SEQ ID NO: 20 and a CDR39 having the amino acid sequence of SEQ ID NO: 29; (vi)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 6 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 12, A CDR2 having the amino acid sequence of SEQ ID NO: 20 and a CDR3 having the amino acid sequence of SEQ ID NO: 30; (vii)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 5 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 10, A CDR2 having the amino acid sequence of SEQ ID NO: 21 and a CDR3 having the amino acid sequence of SEQ ID NO: 31; (viii)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 5 and a CDR3 having the amino acid sequence of SEQ ID NO: 7, and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 13, A CDR2 having the amino acid sequence of SEQ ID NO: 22 and a CDR3 having the amino acid sequence of SEQ ID NO: 32; (ix)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 5 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 12, A CDR2 having the amino acid sequence of SEQ ID NO: 20 and a CDR3 having the amino acid sequence of SEQ ID NO: 30; (x)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: a CDR1 having the amino acid sequence of SEQ ID NO: 14, A CDR2 having the amino acid sequence of SEQ ID NO: 23 and a CDR3 having the amino acid sequence of SEQ ID NO: 33; (xi)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 8, A CDR2 having the amino acid sequence of SEQ ID NO: 24 and A CDR3 having the amino acid sequence of SEQ ID NO: 34; or (xii)(a) a heavy chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 2, A CDR2 having the amino acid sequence of SEQ ID NO: 5 and a CDR3 having the amino acid sequence of SEQ ID NO: 7; and (b) a light chain variable domain comprising: A CDR1 having the amino acid sequence of SEQ ID NO: 15, A CDR2 having the amino acid sequence of SEQ ID NO: 17 and A CDR3 having the amino acid sequence of SEQ ID NO:
35.
13. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the heavy chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, or SEQ ID NO:
43.
14. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, wherein the light chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, or SEQ ID NO:
54.
15. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein: (a) the heavy chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42 or SEQ ID NO:43; and (b) the light chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53 or SEQ ID NO:
54.
16. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 15, wherein: (i)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:36, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 44; (ii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 37, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 45; (iii)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 38, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 46; (iv)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:39, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 47; (v)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:40, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 48; (vi)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:41, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 49; (vii) (a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 40, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 50; (viii)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:40, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 51; (ix)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:40, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 49; (x)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:42, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 52; (xi)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:43, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 53; or (xii)(a) the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:40, and (b) the light chain variable domain comprises the amino acid sequence of SEQ ID NO:
54.
17. The isolated monoclonal antibody of any one of claims 1 to 16, wherein the antibody that specifically binds BCMA is a full-length IgG antibody.
18. The isolated monoclonal antibody of claim 17, wherein the full-length IgG antibody is of human IgG1, IgG2, IgG3 or IgG4 isotype.
19. The isolated monoclonal antibody of claim 18, wherein the full-length IgG antibody is of the human IgG1 isotype.
20. The isolated monoclonal antibody according to claim 19, wherein the antibody comprises the mutations L234A and L235A according to EU numbering of amino acids in the CH2 region.
21. The isolated monoclonal antibody of claim 1, comprising a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61 or SEQ ID NO:
62.
22. The isolated monoclonal antibody of claim 1, comprising a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, or SEQ ID NO:
73.
23. The isolated monoclonal antibody of claim 1, comprising: (i)(a) a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61 or SEQ ID NO:62, and (b) a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 or SEQ ID NO:
73.
24. The isolated monoclonal antibody of claim 23, comprising: (i)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:55, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 63; (ii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 56, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 64; (iii) (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 57, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 65; (iv)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 58, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 66; (v)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 59, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 67; (vi)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 60, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 68; (vii)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 69; (viii)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 70; (ix)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 68, (x)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 61, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 71; (xi)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 62, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 72; or (xii)(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:59, and (b) a light chain comprising the amino acid sequence of SEQ ID NO:
73.
25. An isolated nucleic acid encoding the antibody or antigen-binding fragment thereof according to any one of claims 1 to 24.
26. The isolated nucleic acid of claim 25, wherein the nucleic acid is DNA.
27. An expression vector comprising the nucleic acid according to any one of claims 25 to 26.
28. A method for producing a host cell to produce the antibody or antigen-binding fragment thereof according to any one of claims 1 to 24, comprising transforming the cell with the vector according to claim 27.
29. A host cell for producing the antibody or antigen-binding fragment thereof according to any one of claims 1 to 24, comprising the nucleic acid according to any one of claims 25 to 26.
30. A method for producing an antibody or antigen-binding fragment thereof according to any one of claims 1 to 24, comprising culturing the host cell according to claim 29 in a growth medium under conditions sufficient to produce the antibody, and then isolating and purifying the resulting antibody if necessary.
31. A pharmaceutical composition for treating a BCMA-mediated disease or condition, comprising a therapeutically effective amount of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 24 and one or more pharmaceutically acceptable excipients.
32. The pharmaceutical composition of claim 31, wherein the BCMA-mediated disease or condition is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
33. A pharmaceutical composition for treating a BCMA-mediated disease or condition, comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 24 and at least one other therapeutically active compound.
34. The pharmaceutical composition of claim 33, wherein the BCMA-mediated disease or condition is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
35. The pharmaceutical composition of any one of claims 33 to 34, wherein the other therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent, or a combination thereof.
36. A method for treating a BCMA-mediated disease or condition comprising administering to a subject in need of such treatment an antibody or antigen-binding fragment thereof according to any one of claims 1 to 24 or a pharmaceutical composition according to any one of claims 31 to 35 in a therapeutically effective amount.
37. The method for treating a disease or condition according to claim 36, wherein the BCMA-mediated disease or condition is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
38. A method for treating a BCMA-mediated disease or condition comprising administering to a subject in need of such treatment an antibody or antigen-binding fragment thereof according to any one of claims 1 to 24 and at least one other therapeutically active compound in therapeutically effective amounts.
39. The method for treating a disease or condition according to claim 38, wherein the BCMA-mediated disease or condition is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
40. The method for treating a disease or condition according to any one of claims 38 to 39, wherein the other therapeutically active compound is an antibody, a small molecule, a hormonal therapeutic agent, or a combination thereof.
41. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 24 or the pharmaceutical composition according to any one of claims 31 to 35 for treating a BCMA-mediated disease or disorder in a subject in need of treatment.
42. The use according to claim 41, wherein the BCMA-mediated disease or condition is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
43. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 24 and at least one other therapeutically active compound for treating a BCMA-mediated disease or disorder.
44. The use according to claim 43, wherein the BCMA-mediated disease or condition is selected from: multiple myeloma, chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma.
45. The use according to claim 43, wherein the other therapeutically active compound is a small molecule, a hormonal therapeutic agent or any combination thereof.
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