CN112778417B

CN112778417B - Isolated antigen BCMA-binding protein and use thereof

Info

Publication number: CN112778417B
Application number: CN202011232311.2A
Authority: CN
Inventors: 何晓文; 周金财; 李霄培; 杨月; 史中军; 王华菁; 陈思晔; 杨焕凤
Original assignee: Oricell Therapeutics Co Ltd
Current assignee: Oricell Therapeutics Co Ltd
Priority date: 2019-11-07
Filing date: 2020-11-06
Publication date: 2024-04-12
Anticipated expiration: 2040-11-06
Also published as: CN112778417A

Abstract

The present application relates to an isolated antigen binding protein comprising at least one CDR in a VH having an amino acid sequence as shown in SEQ ID NO. 30 or 59; and which comprises at least one CDR in the VL having the amino acid sequence shown in SEQ ID NO. 26. The present application also relates to nucleic acids encoding the isolated antigen binding proteins, vectors comprising the isolated antigen binding proteins, cells comprising the nucleic acids or the vectors, methods of preparing the isolated antigen binding proteins, and uses of the isolated antigen binding proteins.

Description

Isolated antigen BCMA-binding protein and use thereof

Technical Field

The application relates to the field of biological medicine, in particular to an isolated antigen BCMA-binding protein and application thereof.

Background

A tumor is a disease that severely threatens human health, where myeloma (also known as plasmacytoma) is a widely compromised malignancy. B cell maturation antigens, also known as BCMA, are members of the tumor necrosis receptor (TNFR) family and are expressed predominantly on terminally differentiated B cells, such as memory B cells and plasma cells, particularly common in myeloma.

In recent years, with the progress of molecular biology, genomics and proteomics, a series of molecular targeted drugs for the treatment of myeloma have been continuously developed. The molecular targeting treatment uses some marker molecules over-expressed by tumor cells as targets, and selects targeted blocking agents to perform effective intervention, so as to achieve the effect of inhibiting tumor growth, progress and metastasis. Compared with three traditional treatment means of operation, radiotherapy and chemotherapy, the molecular targeting treatment can efficiently and selectively kill tumor cells, and reduce the damage to normal tissues. However, the effect of tumor treatment by using the molecular targeting technology is still not satisfactory, and there are many improvements.

Disclosure of Invention

In one aspect, the present application provides an isolated antigen binding protein comprising at least one CDR in a VH of amino acid sequence SEQ ID NO 30 or 59; and which comprises at least one CDR in the VL indicated by the amino acid sequence SEQ ID NO: 26.

In certain embodiments, the isolated antigen binding protein has one or more of the following properties:

1) Can be 7×10 ^-11 M or lower KD with BCMA protein, wherein the KD value is determined by Octet;

2) BCMA protein capable of specifically binding to MM1S myeloma cell surface in FACS assay;

3) Can inhibit tumor growth and/or tumor cell proliferation.

In certain embodiments, the BCMA protein comprises a human BCMA protein.

In certain embodiments, the tumor comprises a BCMA positive tumor.

In certain embodiments, the BCMA positive tumor comprises myeloma.

In certain embodiments, the isolated antigen binding protein comprises HCDR1 in a VH set forth in amino acid sequence SEQ ID NO. 30 or 59.

In certain embodiments, the isolated antigen binding protein comprises HCDR2 in a VH set forth in amino acid sequence SEQ ID NO. 30 or 59.

In certain embodiments, the isolated antigen binding protein comprises HCDR3 in a VH set forth in amino acid sequence SEQ ID NO. 30 or 59.

In certain embodiments, the HCDR1 comprises the amino acid sequence set forth in SEQ ID NO. 4.

In certain embodiments, the HCDR2 comprises the amino acid sequence set forth in SEQ ID NO. 12 or 53.

In certain embodiments, the HCDR2 comprises an amino acid sequence set forth in any one of SEQ ID NOs 5, 8 and 52.

In certain embodiments, the HCDR3 comprises the amino acid sequence set forth in SEQ ID NO. 6.

In certain embodiments, the isolated antigen binding protein comprises LCDR1 in a VL set forth in amino acid sequence SEQ ID NO. 26.

In certain embodiments, the isolated antigen binding protein comprises LCDR2 in a VL represented by amino acid sequence SEQ ID NO. 26.

In certain embodiments, the isolated antigen binding protein comprises LCDR3 in a VL set forth in amino acid sequence SEQ ID NO. 26.

In certain embodiments, the LCDR1 comprises the amino acid sequence set forth in SEQ ID NO. 10.

In certain embodiments, the LCDR1 comprises an amino acid sequence set forth in any one of SEQ ID NOs 1 and 7.

In certain embodiments, the LCDR2 comprises the amino acid sequence set forth in SEQ ID NO. 11.

In certain embodiments, the LCDR2 comprises an amino acid sequence set forth in any one of SEQ ID NOs 2 and 9.

In certain embodiments, the LCDR3 comprises the amino acid sequence set forth in SEQ ID NO. 3.

In certain embodiments, the isolated antigen binding protein comprises an antibody or antigen binding fragment thereof.

In certain embodiments, the antigen binding fragment comprises a Fab, fab ', F (ab) 2, fv fragment, F (ab') 2, scFv, di-scFv, and/or dAb.

In certain embodiments, the VL comprises framework regions L-FR1, L-FR2, L-FR3, and L-FR4.

In certain embodiments, the C-terminus of the L-FR1 is directly or indirectly linked to the N-terminus of the LCDR1, and the L-FR1 comprises the amino acid sequence shown in SEQ ID NO. 13.

In certain embodiments, the L-FR1 comprises the amino acid sequence shown in SEQ ID NO. 13.

In certain embodiments, the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 comprises the amino acid sequence shown in SEQ ID NO. 14.

In certain embodiments, the L-FR2 comprises the amino acid sequence shown in SEQ ID NO. 14.

In certain embodiments, the L-FR3 is located between the LCDR2 and the LCDR3, and the L-FR3 comprises the amino acid sequence shown in SEQ ID NO. 15.

In certain embodiments, the L-FR3 comprises the amino acid sequence shown in SEQ ID NO. 15.

In certain embodiments, the N-terminus of the L-FR4 is linked to the C-terminus of the LCDR3 and the L-FR4 comprises the amino acid sequence shown in SEQ ID NO. 16.

In certain embodiments, the L-FR4 comprises the amino acid sequence shown in SEQ ID NO. 16.

In certain embodiments, the VL comprises an amino acid sequence set forth in any one of SEQ ID NOS.23-25.

In certain embodiments, the isolated antigen binding protein comprises an antibody light chain constant region, and the antibody light chain constant region comprises a human igκ constant region.

In certain embodiments, the antibody light chain constant region comprises the amino acid sequence set forth in SEQ ID NO. 31.

In certain embodiments, the isolated antigen binding protein comprises an antibody light chain LC, and the LC comprises an amino acid sequence set forth in any one of SEQ ID NOs 33-35.

In certain embodiments, the VH comprises framework regions H-FR1, H-FR2, H-FR3, and H-FR4.

In certain embodiments, the C-terminus of the H-FR1 is directly or indirectly linked to the N-terminus of the HCDR1, and the H-FR1 comprises the amino acid sequence shown in SEQ ID NO. 17.

In certain embodiments, the H-FR1 comprises the amino acid sequence shown in SEQ ID NO. 17 or 54.

In certain embodiments, the H-FR2 is located between the HCDR1 and the HCDR2, and the H-FR2 comprises the amino acid sequence set forth in any one of SEQ ID NOs 18, 21 and 55.

In certain embodiments, the H-FR2 comprises an amino acid sequence set forth in any one of SEQ ID NOs 18, 21 and 55.

In certain embodiments, the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 comprises the amino acid sequence shown in SEQ ID NO 19 or 56.

In certain embodiments, the H-FR3 comprises the amino acid sequence shown in SEQ ID NO. 19 or 56.

In certain embodiments, the N-terminus of the H-FR4 is linked to the C-terminus of the HCDR3 and the H-FR4 comprises the amino acid sequence shown in SEQ ID NO. 20.

In certain embodiments, the H-FR4 comprises the amino acid sequence shown in SEQ ID NO. 20.

In certain embodiments, the VH comprises an amino acid sequence set forth in any one of SEQ ID NOs 27, 28, 29, 57 and 58.

In certain embodiments, the isolated antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgG heavy chain constant region.

In certain embodiments, the isolated antigen binding protein comprises an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgG1 heavy chain constant region.

In certain embodiments, the antibody heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO. 32.

In certain embodiments, the isolated antigen binding protein comprises an antibody heavy chain HC, and the HC comprises the amino acid sequence set forth in any one of SEQ ID NOs 36, 37, 38, 60 and 61.

In another aspect, the present application provides an isolated nucleic acid molecule or molecules encoding an isolated antigen binding protein described herein.

In another aspect, the present application provides a vector comprising a nucleic acid molecule as described herein.

In another aspect, the present application provides a cell comprising a nucleic acid molecule described herein or a vector described herein.

In another aspect, the present application provides a method of making an isolated antigen binding protein described herein, the method comprising culturing a cell described herein under conditions such that the isolated antigen binding protein described herein is expressed.

In another aspect, the present application provides a pharmaceutical composition comprising an isolated antigen binding protein described herein, a nucleic acid molecule described herein, a vector described herein, and/or a cell described herein, and optionally a pharmaceutically acceptable adjuvant.

In another aspect, the present application provides the use of an isolated antigen binding protein described herein, a nucleic acid molecule described herein, a vector described herein, a cell described herein and/or a pharmaceutical composition described herein in the manufacture of a medicament for the prevention, alleviation and/or treatment of a tumor.

In certain embodiments, the tumor comprises myeloma.

In another aspect, the present application provides a method of preventing, alleviating or treating a tumor, the method comprising administering to a subject in need thereof an isolated antigen binding protein described herein.

In certain embodiments, the tumor comprises myeloma.

In another aspect, the present application provides a method of detecting BCMA in a sample comprising administering the isolated antigen binding protein described herein.

Other aspects and advantages of the present application will become readily apparent to those skilled in the art from the following detailed description. Only exemplary embodiments of the present application are shown and described in the following detailed description. As those skilled in the art will recognize, the present disclosure enables one skilled in the art to make modifications to the disclosed embodiments without departing from the spirit and scope of the invention as described herein. Accordingly, the drawings and descriptions herein are to be regarded as illustrative in nature and not as restrictive.

Drawings

The specific features of the invention related to this application are set forth in the appended claims. The features and advantages of the invention that are related to the present application will be better understood by reference to the exemplary embodiments and the drawings that are described in detail below. The brief description of the drawings is as follows:

FIG. 1 shows the results of detection of binding activity of whole antibodies to cell surface BCMA by 8A7, 8A10, 8A11, hBCMA19, hBCMA22, hBCMA 23;

FIG. 2 shows the results of ADCC activity assays for whole antibodies 8A7, 8A10, 8A11, hBCMA19, hBCMA22, hBCMA 23;

FIG. 3 shows the results of CDC activity assays for 8A10, hBCMA22, hBCMA23 whole antibodies;

FIG. 4 shows the results of an hBCMA22 whole antibody tumor burden experiment;

figure 5 shows the tumor growth inhibition results for hbma 22 whole antibody.

Detailed Description

Further advantages and effects of the invention of the present application will become apparent to those skilled in the art from the disclosure of the present application, from the following description of specific embodiments.

Definition of terms

The present application is further described below: in the present invention, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art. Also, protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, immunology-related terms and laboratory procedures as used herein are terms and conventional procedures that are widely used in the corresponding arts. Meanwhile, in order to better understand the present invention, definitions and explanations of related terms are provided below.

In this application, the term "isolated" generally refers to those obtained from a natural state by artificial means. If a "isolated" substance or component occurs in nature, it may be that the natural environment in which it is located is altered, or that the substance is isolated from the natural environment, or both. For example, a polynucleotide or polypeptide that has not been isolated naturally occurs in a living animal, and the same polynucleotide or polypeptide that has been isolated from the natural state and is of high purity is said to be isolated. The term "isolated" does not exclude the incorporation of artificial or synthetic substances, nor the presence of other impure substances that do not affect the activity of the substance.

In the present application, the term "isolated antigen binding protein" generally refers to a protein having antigen binding ability obtained from a natural state by artificial means. The "isolated antigen binding protein" may comprise an antigen-binding moiety and optionally, a scaffold or framework moiety that allows the antigen-binding moiety to adopt a conformation that promotes binding of the antigen by the antigen-binding moiety. The antigen binding protein may comprise, for example, an antibody-derived protein scaffold or an alternative protein scaffold or artificial scaffold with grafted CDRs or CDR derivatives. Such scaffolds include, but are not limited to, scaffolds comprising antibody sources that are introduced, for example, to stabilize mutations in the three-dimensional structure of the antigen binding protein, as well as fully synthetic scaffolds comprising, for example, biocompatible polymers. See, e.g., korndorfer et al, 2003, proteins: structure, function, andBioiInformatics, 53 (1): 121-129 (2003); roque et al, biotechnol. Prog.20:639-654 (2004). In addition, peptide antibody mimetics ("PAMs") and scaffolds based on antibody mimetics using fibronectin components may be used as scaffolds.

In this application, the term "KD" (likewise, "KD" or "KD") generally refers to "affinity constant" or "equilibrium dissociation constant" and refers to the value obtained at equilibrium, or by dividing the dissociation rate constant (KD) by the association rate constant (ka) in a titration measurement. Binding affinity of a binding protein (e.g., an isolated antigen binding protein described herein) to an antigen (e.g., BCMA protein) is expressed using a binding rate constant (ka), a dissociation rate constant (KD), and a equilibrium dissociation constant (KD). Methods for determining the association and dissociation rate constants are well known in the art. The use of fluorescence-based techniques provides high sensitivity and the ability to examine samples at equilibrium in physiological buffers. For example, the KD values can be determined by Octet, but other experimental approaches and instruments such as BIAcore (biomolecular interaction analysis) assays (e.g., instruments available from BIAcoreInternationalAB, aGEHealthcarecompany, uppsala, sweden) can also be used. In addition, the KD values can also be determined using KinExA (kinetic exclusion assay) available from sapidynes instruments (bose, idaho).

In the present application, the term "BCMA" generally refers to B cell maturation antigen, also known as TNFRSF17, BCM or CD269, which is a member of the tumor necrosis receptor (TNFR) family and is mainly expressed on terminally differentiated B cells, such as memory B cells and plasma cells. BCMA is present in a variety of tumors, particularly in myeloma. Its ligands are known as B cell activators of the TNF family (BAFF) and proliferation-inducing ligands (APRIL). BCMA is involved in mediating plasma cell survival to maintain long-term humoral immunity. The gene for BCMA encodes on chromosome 16, resulting in a primary mRNA transcript of 994 nucleotides in length (NCBI accession No. nm_ 001192.2) which encodes a 184 amino acid protein (np_ 001183.2). In this application, "BCMA" may comprise mutated proteins, such as point mutations, fragments, insertions, deletions and splice variants of full-length wild-type BCMA. Furthermore, in the present application, BCMA protein may comprise human BCMA protein.

In this application, the term "specific binding" or "specific" generally refers to a measurable and reproducible interaction, such as binding between a target and an antibody, that can determine the presence of a target in the presence of a heterogeneous population of molecules (including biomolecules). For example, an antibody that specifically binds a target (which may be an epitope) is one that binds the target with greater affinity, avidity, more readily, and/or for a greater duration than it binds other targets. In one embodiment, the extent of antibody binding to an unrelated target is less than about 10% of the binding of the antibody to the target, as measured, for example, by Radioimmunoassay (RIA). For example, in the present application, the isolated antigen binding protein is capable of binding to BCMA protein with a dissociation constant (KD) of <7x10 "11M or less. In certain embodiments, the antibodies specifically bind to epitopes on proteins that are conserved among proteins of different species. In another embodiment, specific binding may include, but is not required to be, exclusively binding.

In this application, the term "inhibit" generally refers to reducing the growth rate of a cell or the number of cells. For example, the isolated antigen binding proteins described herein are capable of inhibiting tumor growth and/or tumor cell proliferation.

In this application, the term "tumor" generally refers to a neoplasm or solid lesion formed by abnormal cell growth. In this application, the tumor may be a solid tumor or a hematological tumor. For example, in the present application, a tumor may be a BCMA positive tumor, wherein the BCMA positive tumor may include myeloma.

In this application, the term "variable domain" generally refers to the amino-terminal domain of an antibody heavy or light chain. The variable domains of the heavy and light chains may be referred to as "VH" and "VL", respectively. These domains are typically the most variable portions of an antibody (relative to other antibodies of the same type) and comprise antigen binding sites.

In the present application, the term "variable" generally refers to the fact that there is a large difference in sequence in certain segments of the variable domain between antibodies. The V domain mediates antigen binding and determines the specificity of a particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains. Instead, it concentrates in three segments called hypervariable regions (CDRs or HVRs) in the light and heavy chain variable domains. The more highly conserved parts of the variable domains are called Framework Regions (FR). The variable domains of the natural heavy and light chains each comprise four FR regions, mostly in a β -sheet configuration, connected by three CDRs, which form a circular connection and in some cases form part of a β -sheet structure. The CDRs in each chain are held together in close proximity by the FR regions, and the CDRs from the other chain together promote the formation of the antigen binding site of the antibody (see Kabat et al, sequences of Immunological Interest, fifth Edition, national Institute of Health, bethesda, md. (1991)). The constant domains are not directly involved in binding of antibodies to antigens, but exhibit various effector functions, such as antibody involvement in antibody-dependent cellular cytotoxicity.

In this application, the term "antibody" generally refers to an immunoglobulin or fragment or derivative thereof, and encompasses any polypeptide comprising an antigen binding site, whether produced in vitro or in vivo. The term includes, but is not limited to, polyclonal, monoclonal, monospecific, multispecific, nonspecific, humanized, single chain, chimeric, synthetic, recombinant, hybrid, mutant, and grafted antibodies. Unless otherwise modified by the term "intact", as in "intact antibodies", for the purposes of the present invention, the term "antibody" also includes antibody fragments such as Fab, F (ab') 2, fv, scFv, fd, dAb, and other antibody fragments that retain antigen binding function (e.g., specifically bind BCMA). Typically, such fragments should include an antigen binding domain. The basic 4-chain antibody unit is a heterotetrameric glycoprotein consisting of two identical light (L) chains and two identical heavy (H) chains. IgM antibodies consist of 5 basic heterotetramer units with another polypeptide called the J chain and contain 10 antigen binding sites, whereas IgA antibodies comprise 2-5 basic 4-chain units that can polymerize in conjunction with the J chain to form multivalent combinations. In the case of IgG, the 4-chain unit is typically about 150,000 daltons. Each L chain is linked to the H chain by one covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has a variable domain (VH) at the N-terminus, followed by three constant domains (CH) for each of the alpha and gamma chains, followed by four CH domains for the mu and epsilon isoforms. Each L chain has a variable domain (VL) at the N-terminus and a constant domain at its other end. VL corresponds to VH, and CL corresponds to the first constant domain of the heavy chain (CH 1). Specific amino acid residues are believed to form an interface between the light chain and heavy chain variable domains. The VH and VL pairs together form a single antigen binding site. For the structure and properties of different classes of antibodies, see, e.g., basic and Clinical Immunology,8th Edition,Daniel P.Sties,Abba I.Terr and Tristram G.Parsolw (eds), appleton & Lange, norwalk, conn, 1994, page 71 and chapter 6. L chains from any vertebrate species can be divided into one of two distinct types, termed kappa and lambda, based on the amino acid sequence of their constant domains. Immunoglobulins can be assigned to different classes or isotypes depending on the amino acid sequence of their heavy Chain (CH) constant domain. There are five classes of immunoglobulins: igA, igD, igE, igG and IgM, have heavy chains named α, δ, ε, γ and μ, respectively. Based on the relatively small differences in CH sequence and function, the γ and α classes are further divided into subclasses, e.g., humans express the following subclasses: igG1, igG2A, igG2B, igG3, igG4, igA1 and IgK1.

In this application, the term "CDR" generally refers to a region of an antibody variable domain whose sequence is highly variable and/or forms a structurally defined loop. Typically, an antibody comprises six CDRs; three in VH (HCDR 1, HCDR2, HCDR 3), and three in VL (LCDR 1, LCDR2, LCDR 3). In natural antibodies, HCDR3 and LCDR3 show most of the diversity of the six CDRs, and in particular HCDR3 is thought to play a unique role in conferring fine specificity to antibodies. See, e.g., xu et al, immunity 13:37-45 (2000); johnson and Wu, in Methods in Molecular Biology 248:1-25 (Lo, ed., human Press, totowa, N.J., 2003). In fact, naturally occurring camelid antibodies consisting of heavy chains only function normally and stably in the absence of light chains. See, e.g., hamers-Casterman et al, nature 363:446-448 (1993); sheiff et al, nature Structure. Biol.3:733-736 (1996).

In this application, the term "FR" generally refers to the more highly conserved portion of the antibody variable domain, which is referred to as the framework region. Typically, the variable domains of the natural heavy and light chains each comprise four FR regions, namely four in VH (H-FR 1, H-FR2, H-FR3, and H-FR 4), and four in VL (L-FR 1, L-FR2, L-FR3, and L-FR 4). For example, VL of an isolated antigen binding protein described herein may comprise framework regions L-FR1, L-FR2, L-FR3, and L-FR4. The VH of the isolated antigen binding proteins described herein may include framework regions H-FR1, H-FR2, H-FR3, and H-FR4.

In the present application, the term "antigen binding fragment" generally refers to a fragment having antigen binding activity. In the present application, the antigen binding fragment may comprise a Fab, fab ', F (ab) 2, fv fragment, F (ab') 2, scFv, di-scFv and/or dAb.

In this application, the term "directly coupled" is used to refer to a direct connection as opposed to the term "indirectly coupled". For example, the direct linkage may be where there is no spacer between the substances. The spacer may be a linker. For example, the linker may be a peptide linker. The term "indirect linkage" generally refers to the situation where the materials are not directly linked. For example, the indirect connection may be the case where the connection is through a spacer. For example, in the isolated antigen binding proteins described herein, the C-terminus of the L-FR1 and the N-terminus of the LCDR1 can be directly or indirectly linked.

In the present application, the term "isolated nucleic acid molecule" generally refers to an isolated form of nucleotides, deoxyribonucleotides or ribonucleotides of any length, or an analogue isolated from its natural environment or synthesized synthetically.

In the present application, the term "vector" generally refers to a nucleic acid vector into which a polynucleotide encoding a protein can be inserted and the protein expressed. The vector may be expressed by transforming, transducing or transfecting a host cell such that the genetic element carried thereby is expressed within the host cell. For example, the carrier comprises: a plasmid; phagemid; a cosmid; artificial chromosomes such as Yeast Artificial Chromosome (YAC), bacterial Artificial Chromosome (BAC) or P1-derived artificial chromosome (PAC); phages such as lambda phage or M13 phage, animal viruses, etc. Animal virus species used as vectors are retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpesviruses (e.g., herpes simplex viruses), poxviruses, baculoviruses, papillomaviruses, papilloma-virus-papilloma-vacuolated viruses (e.g., SV 40). A vector may contain a variety of elements for controlling expression, including promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. In addition, the vector may also contain a replication origin. It is also possible for the vector to include components that assist it in entering the cell, such as viral particles, liposomes or protein shells, but not just these.

In this application, the term "cell" generally refers to a single cell, cell line or cell culture that may or may not be the recipient of a subject plasmid or vector, which includes a nucleic acid molecule of the invention or a vector of the invention. Cells may include progeny of a single cell. The offspring may not necessarily be identical to the original parent cell (either in the form of the total DNA complement or in the genome) due to natural, accidental or deliberate mutation. Cells may include cells transfected in vitro with the vectors of the invention. The cell may be a bacterial cell (e.g., E.coli), a yeast cell, or other eukaryotic cell, such as COS cells, chinese Hamster Ovary (CHO) cells, heLa cells, HEK293 cells, COS-1 cells, NS0 cells, or myeloma cells. In certain embodiments, the cell is a mammalian cell. In certain embodiments, the mammalian cell is a HEK293 cell.

In this application, the term "pharmaceutical composition" generally refers to a composition suitable for administration to a patient, preferably a human patient. For example, a pharmaceutical composition described herein may comprise an isolated antigen binding protein described herein, a nucleic acid molecule described herein, a vector described herein, and/or a cell described herein, and optionally a pharmaceutically acceptable adjuvant. In addition, the pharmaceutical composition may further comprise one or more (pharmaceutically effective) suitable formulations of carriers, stabilizers, excipients, diluents, solubilizers, surfactants, emulsifiers and/or preservatives. The acceptable ingredients of the composition are preferably non-toxic to the recipient at the dosages and concentrations employed. Pharmaceutical compositions of the invention include, but are not limited to, liquid, frozen and lyophilized compositions.

In this application, the term "pharmaceutically acceptable adjuvant" generally refers to any and all solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration, are generally safe, nontoxic, and neither biologically nor otherwise undesirable.

In this application, the term "subject" generally refers to a human or non-human animal, including but not limited to, cats, dogs, horses, pigs, cows, sheep, rabbits, mice, rats, or monkeys.

In this application, the term "comprising" is generally intended to include the features specifically recited, but does not exclude other elements.

In this application, the term "about" generally means ranging from 0.5% to 10% above or below the specified value, e.g., ranging from 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below the specified value.

Detailed Description

Isolated antigen binding proteins

In one aspect the present application provides an isolated antigen binding protein comprising at least one CDR in a VH having an amino acid sequence as shown in SEQ ID NO. 30 or 59; and which comprises at least one CDR in the VL having the amino acid sequence shown in SEQ ID NO. 26.

For example, the VH of an isolated antigen binding protein described herein may comprise an amino acid sequence as set out in any one of SEQ ID NOs 27, 28, 29, 57 and 58.

For example, the VL of the isolated antigen-binding proteins described herein may comprise the amino acid sequence set forth in any one of SEQ ID NOS.23-25.

For example, in the present application, the isolated antigen binding protein may comprise HCDR1 in a VH having an amino acid sequence as set forth in SEQ ID NO. 30 or 59. For example, in the present application, the isolated antigen binding protein may comprise HCDR2 in a VH having an amino acid sequence as set forth in SEQ ID NO. 30 or 59. For example, in the present application, the isolated antigen binding protein may comprise HCDR3 in a VH having an amino acid sequence as set forth in SEQ ID NO. 30 or 59. For another example, in the present application, the isolated antigen binding protein may comprise LCDR1 in VL having the amino acid sequence shown in SEQ ID NO. 26. For example, in the present application, the isolated antigen binding protein may comprise LCDR2 in VL having the amino acid sequence shown in SEQ ID NO. 26. For example, in the present application, the isolated antigen binding protein may comprise LCDR3 in VL having the amino acid sequence shown in SEQ ID NO. 26.

Properties of the isolated antigen binding proteins

In the present application, the isolated antigen binding protein may have one or more of the following properties:

3) Can inhibit tumor growth and/or tumor cell proliferation.

In the present application, the isolated antigen binding protein can be present in a 7X 10 ratio ^-11 M or lower K _D Binding to BCMA protein, wherein the K _D The value can be determined by Octet. For example, the isolated antigen binding proteins described herein bind to K of human-derived BCMA protein _D The value may be 7×10 or less ^-11 M、≤6.5×10 ^-11 M、≤6×10 ^-11 M、≤5.5×10 ^-11 M、≤5×10 ^-11 M、≤4.5×10 ^-11 M、≤4×10 ^-11 M、≤3.5×10 ^-11 M、≤3×10 ^-11 M、≤2.5×10 ^-11 M、≤2×10 ^-11 M、≤1.5×10 ^-11 M、≤1×10 ^-11 M. For another example, an isolated antigen binding protein described herein binds to K of a murine-derived BCMA protein _D The value may be 7×10 or less ^-11 M、≤6.5×10 ^-11 M、≤6×10 ^-11 M、≤5.5×10 ^-11 M、≤5×10 ^- ¹¹ M、≤4.5×10 ^-11 M、≤4×10 ^-11 M、≤3.5×10 ^-11 M、≤3×10 ^-11 M、≤2.5×10 ^-11 M、≤2×10 ^-11 M、≤1.5×10 ^-11 M、≤1×10 ^-11 M. For another example, an isolated antigen binding protein described herein binds to K of a monkey-derived BCMA protein _D The value may be 7×10 or less ^-11 M、≤6.5×10 ^-11 M、≤6×10 ^-11 M、≤5.5×10 ^-11 M、≤5×10 ^-11 M、≤4.5×10 ^-11 M、≤4×10 ^-11 M、≤3.5×10 ^-11 M、≤3×10 ^-11 M、≤2.5×10 ^-11 M、≤2×10 ^-11 M、≤1.5×10 ^-11 M、≤1×10 ^-11 M。

In this application, the KD value can also be determined by ELISA, competition ELISA or BIACORE or KINEXA.

In the present application, the isolated antigen binding protein is capable of specifically binding to BCMA protein on the surface of MM1S myeloma cells, which specific binding can be determined by FACS. For example, the specific binding of the isolated antigen binding proteins described herein to BCMA proteins on the surface of MM1S myeloma cells can be reflected by the half maximal effector concentration (EC 50) in FACS assays, e.g., lower half maximal effector concentration (EC 50) indicates better specific binding. For example, the isolated antigen binding protein may have an EC50 value of 0.01 μg/ml to 0.10 μg/ml, 0.01 μg/ml to 0.15 μg/ml, 0.01 μg/ml to 0.20 μg/ml, 0.01 μg/ml to 0.25 μg/ml, 0.01 μg/ml to 0.30 μg/ml, 0.01 μg/ml to 0.35 μg/ml, 0.01 μg/ml to 0.40 μg/ml, 0.01 μg/ml to 0.45 μg/ml, or 0.01 μg/ml to 0.50 μg/ml for binding to BCMA protein on the surface of MM1S myeloma cells in a FACS assay.

In the present application, the isolated antigen binding proteins described herein are capable of inhibiting tumor growth and/or tumor cell proliferation, e.g., capable of reducing tumor volume by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 99%, or 100%. For another example, the number of tumor cells can be reduced by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 99%, or 100%.

In the present application, the BCMA protein may comprise a human BCMA protein, which may comprise GeneBank accession No.: the amino acid sequence shown in BAB 60895.1.

In this application, the tumor may comprise a BCMA positive tumor, which may comprise bone marrow cancer.

In the present application, the isolated antigen binding protein is capable of inhibiting the growth of tumor cells by inducing antibody dependent cell-mediated cytotoxicity (ADCC). For example, the isolated antigen binding proteins described herein are capable of bringing cytotoxic infiltrating T lymphocytes into tumor tissue, thereby inhibiting the growth of tumor cells. For another example, the isolated antigen binding proteins described herein can also indirectly inhibit the proliferation of BCMA positive cells by macrophage-related mechanisms.

In this application, the isolated antigen binding protein has a certain endocytic activity. The endocytic activity can be detected by the Confocal method.

The isolated antigen bindingSpecies of proteins

In the present application, the isolated antigen binding protein may comprise an antibody or antigen binding fragment thereof. For example, isolated antigen binding proteins described herein may include, but are not limited to, recombinant antibodies, monoclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, bispecific antibodies, single chain antibodies, diabodies, triabodies, tetrabodies, fv fragments, scFv fragments, fab 'fragments, F (ab') 2 fragments, and camelized single domain antibodies.

In the present application, the antibody may be a humanized antibody. In other words, the isolated antigen binding proteins described herein may be antibodies or variants, derivatives, analogs or fragments thereof that immunospecifically bind to a related antigen (e.g., human BCMA) and that comprise a Framework (FR) region having substantially the amino acid sequence of a human antibody and a Complementarity Determining Region (CDR) having substantially the amino acid sequence of a non-human antibody. "substantially" herein in the context of a CDR means that the amino acid sequence of the CDR is at least 80%, preferably at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the amino acid sequence of a CDR of a non-human antibody. The humanized antibody may comprise substantially all of at least one and typically two variable domains (Fab, fab ', F (ab') 2, fabC, fv) in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin (i.e., an antibody) and all or substantially all of the framework regions are those having a human immunoglobulin consensus sequence. Preferably, the humanized antibody further comprises at least a portion of an immunoglobulin constant region (e.g., an Fc), typically that of a human immunoglobulin. In some embodiments, the humanized antibody comprises at least a variable domain of a light chain and a heavy chain. Antibodies may also include CH1, hinge, CH2, CH3, and CH4 regions of the heavy chain. In some embodiments, the humanized antibody comprises only humanized light chains. In some embodiments, the humanized antibody comprises only a humanized heavy chain. In particular embodiments, the humanized antibody comprises only a humanized variable domain of a light chain and/or a humanized heavy chain.

In the present application, the antigen binding fragment may comprise a Fab, fab ', F (ab) 2, fv fragment, F (ab') 2, scFv, di-scFv and/or dAb.

CDR

In this application, the LCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 10.

In the present application, the LCDR1 may comprise an amino acid sequence set forth in any one of SEQ ID NOs 1 and 7.

In this application, the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 11.

In the present application, the LCDR2 may comprise an amino acid sequence as set forth in any one of SEQ ID NOs 2 and 9.

In this application, the LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3.

For example, LCDR1 of the isolated antigen binding proteins described herein may comprise the amino acid sequence shown in SEQ ID NO. 1, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 2, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3.

For example, LCDR1 of an isolated antigen binding protein described herein may comprise the amino acid sequence shown in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 2, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3.

For example, LCDR1 of an isolated antigen binding protein described herein may comprise the amino acid sequence shown in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 9, and LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3.

In the present application, the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 4.

In the present application, the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 12 or 53.

In the present application, the HCDR2 may comprise the amino acid sequence set forth in any one of SEQ ID NOs 5, 8 and 52.

In the present application, the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 6.

For example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 5, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

For example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 8, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

For example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 52, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

For another example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 1, LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 2, LCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 5, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

For another example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 2, LCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 8, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6. For another example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 9, LCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 5, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

For another example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 2, LCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 52, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

For another example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 9, LCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 52, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

For another example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 9, LCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence set forth in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence set forth in SEQ ID NO. 8, and HCDR3 may comprise the amino acid sequence set forth in SEQ ID NO. 6.

FR

In the present application, the VL of the isolated antigen binding protein may comprise the framework regions L-FR1, L-FR2, L-FR3, and L-FR4.

In the present application, the L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 13.

For example, the C-terminus of the L-FR1 can be directly or indirectly linked to the N-terminus of the LCDR1, and the L-FR1 can comprise the amino acid sequence shown in SEQ ID NO. 13.

In the present application, the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 14.

For example, the L-FR2 is located between the LCDR1 and the LCDR2, and the L-FR2 can comprise the amino acid sequence shown in SEQ ID NO. 14.

In the present application, the L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 15.

For example, the L-FR3 is located between the LCDR2 and the LCDR3, and the L-FR3 can comprise the amino acid sequence shown in SEQ ID NO. 15.

In the present application, the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16.

For example, the N-terminus of the L-FR4 is linked to the C-terminus of the LCDR3, and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16.

For another example, an isolated antigen binding protein described herein may comprise the amino acid sequence set forth in SEQ ID NO. 13, L-FR2 may comprise the amino acid sequence set forth in SEQ ID NO. 14, L-FR3 may comprise the amino acid sequence set forth in SEQ ID NO. 15, and L-FR4 may comprise the amino acid sequence set forth in SEQ ID NO. 16.

In the present application, the VH of the isolated antigen binding protein may include framework regions H-FR1, H-FR2, H-FR3, and H-FR4.

In the present application, the H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 17 or 54.

In the present application, the C-terminal end of the H-FR1 is directly or indirectly linked to the N-terminal end of the HCDR1, and the H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 17 or 54.

In the present application, the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 22 or 55.

In the present application, the H-FR2 is located between the HCDR1 and the HCDR2, and the H-FR2 may comprise the amino acid sequence shown in any one of SEQ ID NOs 18, 21 and 55.

In the present application, the H-FR2 may comprise the amino acid sequence shown in any one of SEQ ID NOs 18, 21 and 55.

In the present application, the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 19 or 56.

In the present application, the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO 19 or 56.

In the present application, the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20.

In the present application, the N-terminus of the H-FR4 is linked to the C-terminus of the HCDR3, and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20.

For example, the H-FR1 of the isolated antigen binding proteins described herein may comprise the amino acid sequence shown in SEQ ID NO. 17, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 18, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 19, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20.

For example, the H-FR1 of the isolated antigen binding proteins described herein may comprise the amino acid sequence shown in SEQ ID NO. 17, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 21, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 19, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20.

For example, the H-FR1 of the isolated antigen binding proteins described herein can comprise the amino acid sequence shown in SEQ ID NO. 54, H-FR2 can comprise the amino acid sequence shown in SEQ ID NO. 55, H-FR3 can comprise the amino acid sequence shown in SEQ ID NO. 56, and H-FR4 can comprise the amino acid sequence shown in SEQ ID NO. 20.

For another example, the isolated antigen binding protein described herein may have L-FR1 comprising the amino acid sequence shown in SEQ ID NO. 13, L-FR2 comprising the amino acid sequence shown in SEQ ID NO. 14, L-FR3 comprising the amino acid sequence shown in SEQ ID NO. 15, L-FR4 comprising the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 comprising the amino acid sequence shown in SEQ ID NO. 17, H-FR2 comprising the amino acid sequence shown in SEQ ID NO. 18, H-FR3 comprising the amino acid sequence shown in SEQ ID NO. 19, and H-FR4 comprising the amino acid sequence shown in SEQ ID NO. 20.

For another example, the isolated antigen binding protein described herein may have L-FR1 comprising the amino acid sequence shown in SEQ ID NO. 13, L-FR2 comprising the amino acid sequence shown in SEQ ID NO. 14, L-FR3 comprising the amino acid sequence shown in SEQ ID NO. 15, L-FR4 comprising the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 comprising the amino acid sequence shown in SEQ ID NO. 17, H-FR2 comprising the amino acid sequence shown in SEQ ID NO. 21, H-FR3 comprising the amino acid sequence shown in SEQ ID NO. 19, and H-FR4 comprising the amino acid sequence shown in SEQ ID NO. 20.

For another example, the isolated antigen binding protein described herein may have L-FR1 comprising the amino acid sequence shown in SEQ ID NO. 13, L-FR2 comprising the amino acid sequence shown in SEQ ID NO. 14, L-FR3 comprising the amino acid sequence shown in SEQ ID NO. 15, L-FR4 comprising the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 comprising the amino acid sequence shown in SEQ ID NO. 54, H-FR2 comprising the amino acid sequence shown in SEQ ID NO. 55, H-FR3 comprising the amino acid sequence shown in SEQ ID NO. 56, and H-FR4 comprising the amino acid sequence shown in SEQ ID NO. 20.

VL and VH

The isolated antigen binding proteins described herein may comprise an antibody light chain variable region VL and an antibody heavy chain variable region VH. For example, the VL may comprise the amino acid sequence shown in SEQ ID NO. 26 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 30 or 59. For another example, the VL of the isolated antigen-binding protein may comprise an amino acid sequence set forth in any one of SEQ ID NOs 23-25. The VH of the isolated antigen binding protein may comprise an amino acid sequence set out in any one of SEQ ID NOs 27, 28, 29, 57 and 58.

For example, the VL may comprise the amino acid sequence shown in SEQ ID NO. 23 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 27.

For example, the VL may comprise the amino acid sequence shown in SEQ ID NO. 24 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 28.

For example, the VL may comprise the amino acid sequence shown in SEQ ID NO. 25 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 29.

For example, the VL may comprise the amino acid sequence shown in SEQ ID NO. 24 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 57.

For example, the VL may comprise the amino acid sequence shown in SEQ ID NO. 25 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 57.

For example, the VL may comprise the amino acid sequence shown in SEQ ID NO. 25 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 58.

Light and heavy chains

In the present application, the isolated antigen binding protein may comprise an antibody light chain constant region, and the antibody light chain constant region may comprise a human igkappa constant region. For example, the antibody light chain constant region may comprise the amino acid sequence shown in SEQ ID NO. 31.

In the present application, the isolated antigen binding protein may include an antibody heavy chain constant region, and the antibody heavy chain constant region may be derived from a human IgG heavy chain constant region. In certain embodiments, the isolated antigen binding protein may comprise an antibody heavy chain constant region, and the antibody heavy chain constant region may be derived from a human IgG1 heavy chain constant region. For example, the antibody heavy chain constant region may comprise the amino acid sequence shown in SEQ ID NO. 32.

In the present application, the isolated antigen binding protein may comprise an antibody light chain LC, and the LC may comprise an amino acid sequence as set forth in any one of SEQ ID NOs 33 to 35.

In the present application, the isolated antigen binding protein may comprise an antibody heavy chain HC, and the HC may comprise the amino acid sequence set forth in any one of SEQ ID NOs 36, 37, 38, 60 and 61.

The isolated antigen binding proteins described herein may comprise an antibody light chain and an antibody heavy chain.

For example, the light chain may comprise the amino acid sequence shown in SEQ ID NO. 33 and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 36.

For example, the light chain may comprise the amino acid sequence shown in SEQ ID NO. 34 and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 37.

For example, the light chain may comprise the amino acid sequence shown in SEQ ID NO. 35 and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 38.

For example, the light chain may comprise the amino acid sequence shown in SEQ ID NO. 34 and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 64.

For example, the light chain may comprise the amino acid sequence shown in SEQ ID NO. 35 and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 64.

For example, the light chain may comprise the amino acid sequence shown in SEQ ID NO. 35 and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 65.

In the present application, the light chain of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 33, and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 36. Wherein, LCDR1 of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 1, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 2, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 5, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 6. Wherein the isolated antigen binding protein L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 13, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 15, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 17, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 18, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 19, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20. The VL may comprise the amino acid sequence shown in SEQ ID NO. 23, and the VH may comprise the amino acid sequence shown in SEQ ID NO. 27. For example, the isolated antigen binding protein may be an 8A7 whole antibody.

In this application, the light chain of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 34, and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 37. Wherein, LCDR1 of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 2, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 8, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 6. Wherein the isolated antigen binding protein L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 13, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 15, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 17, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 18, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 19, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20. The VL may comprise the amino acid sequence shown in SEQ ID NO. 24, and the VH may comprise the amino acid sequence shown in SEQ ID NO. 28. For example, the isolated antigen binding protein may be an 8a10 whole antibody.

In the present application, the light chain of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 35, and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 38. Wherein, LCDR1 of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 9, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 5, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 6. Wherein the isolated antigen binding protein L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 13, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 15, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 17, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 21, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 19, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20. The VL may comprise the amino acid sequence shown in SEQ ID NO. 25, and the VH may comprise the amino acid sequence shown in SEQ ID NO. 29. For example, the isolated antigen binding protein may be an 8a11 whole antibody.

In this application, the light chain of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 34, and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 60. Wherein, LCDR1 of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 2, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 52, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 6. Wherein the isolated antigen binding protein L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 13, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 15, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 54, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 55, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 56, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20. The VL may comprise the amino acid sequence shown in SEQ ID NO. 24 and the VH may comprise the amino acid sequence shown in SEQ ID NO. 57. For example, the isolated antigen binding protein may be hbma 19 whole antibody.

In the present application, the light chain of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 35, and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 60. Wherein, LCDR1 of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 9, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 52, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 6. Wherein the isolated antigen binding protein L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 13, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 15, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 54, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 55, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 56, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20. The VL may comprise the amino acid sequence shown in SEQ ID NO. 25, and the VH may comprise the amino acid sequence shown in SEQ ID NO. 57. For example, the isolated antigen binding protein may be hbma 22 whole antibody.

In the present application, the light chain of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 35, and the heavy chain may comprise the amino acid sequence shown in SEQ ID NO. 61. Wherein, LCDR1 of the isolated antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO. 7, LCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 9, LCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 3, and HCDR1 may comprise the amino acid sequence shown in SEQ ID NO. 4, HCDR2 may comprise the amino acid sequence shown in SEQ ID NO. 8, and HCDR3 may comprise the amino acid sequence shown in SEQ ID NO. 6. Wherein the isolated antigen binding protein L-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 13, L-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 14, L-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 15, L-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 16, and H-FR1 may comprise the amino acid sequence shown in SEQ ID NO. 54, H-FR2 may comprise the amino acid sequence shown in SEQ ID NO. 55, H-FR3 may comprise the amino acid sequence shown in SEQ ID NO. 56, and H-FR4 may comprise the amino acid sequence shown in SEQ ID NO. 20. The VL may comprise the amino acid sequence shown in SEQ ID NO. 25, and the VH may comprise the amino acid sequence shown in SEQ ID NO. 58. For example, the isolated antigen binding protein may be hbma 23 whole antibody.

Nucleic acid molecules, vectors, cells, methods of preparation and pharmaceutical compositions

In another aspect, the present application also provides an isolated nucleic acid molecule or molecules, which may encode an isolated antigen binding protein as described herein. The isolated nucleic acid molecule or molecules described herein may be any length of isolated form of a nucleotide, deoxyribonucleotide or ribonucleotide, or an analogue thereof either isolated from the natural environment or synthesized, but may encode an isolated antigen binding protein described herein.

In another aspect, the present application also provides vectors, which may comprise the nucleic acid molecules described herein. The vector may be expressed by transforming, transducing or transfecting a host cell such that the genetic element carried thereby is expressed within the host cell. For example, the carrier may comprise: a plasmid; phagemid; a cosmid; artificial chromosomes such as Yeast Artificial Chromosome (YAC), bacterial Artificial Chromosome (BAC) or P1-derived artificial chromosome (PAC); phages such as lambda phage or M13 phage, animal viruses, etc. Animal virus species used as vectors are retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpesviruses (e.g., herpes simplex viruses), poxviruses, baculoviruses, papillomaviruses, papilloma-virus-papilloma-vacuolated viruses (e.g., SV 40). For another example, the vector may contain a variety of elements that control expression, including promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. In addition, the vector may further contain a replication origin. In addition, the vector may include components that assist it in entering the cell, such as viral particles, liposomes, or protein shells, but not exclusively.

In another aspect, the present application also provides a cell, which may comprise a nucleic acid molecule as described herein or a vector as described herein. The cells may include progeny of a single cell. The offspring may not necessarily be identical to the original parent cell (either in the form of the total DNA complement or in the genome) due to natural, accidental or deliberate mutation. In certain embodiments, the cells may also include cells transfected in vitro with the vectors of the invention. In certain embodiments, the cell may be a bacterial cell (e.g., E.coli), a yeast cell, or other eukaryotic cell, such as a COS cell, a Chinese Hamster Ovary (CHO) cell, a HeLa cell, a HEK293 cell, a COS-1 cell, an NS0 cell, or a myeloma cell. In certain embodiments, the cell may be a mammalian cell. In certain embodiments, the mammalian cell may be a HEK293 cell.

In another aspect, the present application also provides methods of making an isolated antigen binding protein described herein, which methods can comprise culturing a cell described herein under conditions such that the isolated antigen binding protein described herein is expressed.

In another aspect, the present application also provides a pharmaceutical composition, which may comprise an isolated antigen binding protein described herein, a nucleic acid molecule described herein, a vector described herein, and/or a cell described herein, and optionally a pharmaceutically acceptable adjuvant.

In certain embodiments, the pharmaceutical compositions may further comprise one or more (pharmaceutically effective) suitable formulations of carriers, stabilizers, excipients, diluents, solubilizers, surfactants, emulsifiers and/or preservatives. The acceptable ingredients of the composition are preferably non-toxic to the recipient at the dosages and concentrations employed. Pharmaceutical compositions of the invention include, but are not limited to, liquid, frozen and lyophilized compositions.

In certain embodiments, the pharmaceutically acceptable adjuvant may include any and all solvents, dispersion media, coatings, isotonic agents, and absorption delaying agents that are compatible with pharmaceutical administration, are generally safe, nontoxic, and neither biologically nor otherwise undesirable.

In certain embodiments, the pharmaceutical composition may comprise parenteral, transdermal, endoluminal, intra-arterial, intrathecal and/or intranasal administration or direct injection into tissue. For example, the pharmaceutical composition may be administered to a patient or subject by infusion or injection. In certain embodiments, the administration of the pharmaceutical composition may be performed by different means, such as intravenous, intraperitoneal, subcutaneous, intramuscular, topical or intradermal administration. In certain embodiments, the pharmaceutical composition may be administered without interruption. The uninterrupted (or continuous) administration may be achieved by a small pump system worn by the patient to measure the therapeutic agent flowing into the patient, as described in WO 2015/036583.

Use and application

In another aspect, the present application also provides the use of an isolated antigen binding protein described herein, a nucleic acid molecule described herein, a vector described herein, a cell described herein and/or a pharmaceutical composition described herein in the manufacture of a medicament for the prevention, alleviation and/or treatment of a tumor.

In another aspect, the present application also provides a method of preventing, alleviating or treating a tumor, which method may comprise administering to a subject in need thereof an isolated antigen binding protein as described herein. In the present application, the administration may be performed in different ways, for example intravenous, intratumoral, intraperitoneal, subcutaneous, intramuscular, topical or intradermal administration.

In another aspect, the isolated antigen binding proteins described herein, the nucleic acid molecules described herein, the vectors described herein, the cells described herein, and/or the pharmaceutical compositions described herein can be used to prevent, ameliorate or treat a tumor.

In this application, the tumor may be a solid tumor or a hematological tumor. For example, the tumor may comprise a BCMA positive tumor, which may comprise myeloma.

In this application, the subject may include humans and non-human animals. For example, the subject may include, but is not limited to, a cat, dog, horse, pig, cow, sheep, rabbit, mouse, rat, or monkey.

In another aspect, the present application also provides a method of detecting BCMA in a sample comprising administering the isolated antigen binding protein described herein. In the present application, the administration may be performed in different ways, for example intravenous, intratumoral, intraperitoneal, subcutaneous, intramuscular, topical or intradermal administration.

Without intending to be limited by any theory, the following examples are meant to illustrate the protein molecules, methods of preparation, uses, and the like of the present application and are not intended to limit the scope of the invention of the present application. Examples do not include detailed descriptions of conventional methods, such as those used to construct vectors and plasmids, methods of inserting genes encoding proteins into such vectors and plasmids, or methods of introducing plasmids into host cells. Such methods are well known to those having ordinary skill in the art and are described in numerous publications, including Sambrook, j., fritsch, e.f. and maniis, t. (1989) Molecular Cloning: a Laboratory Manual,2nd edition,Cold spring Harbor Laboratory Press.

Examples

Example 1 screening of anti-BCMA antibodies Using phage antibody library

The recombinant human BCMA-hFc and BCMA-His proteins (Shanghai original energy cell medicine technology Co., ltd.) are utilized, wherein, human BCMA-hFc represents a protein formed by connecting human BCMA with human Fc, the amino acid sequence of human BCMA protein is shown as GeneBank accession number: BAB60895.1, and the amino acid of human Fc is shown as SEQ ID NO: 51), and phage natural humanized antibody library (Shanghai original energy cell medicine technology Co., ltd.) is alternately sorted, and four rounds of sorting are carried out together. The antigen BCMA-hFc protein was coated in ELISA tube with CBS buffer at 1ml, BCMA-hFc and BCMA-His concentrations of 40. Mu.g/ml (first, second round) or 20. Mu.g/ml (third, fourth round), overnight at 4 ℃; 2ml of PBS buffer containing 5% of skimmed milk powder for the next time is used for sealing an immune tube; adding 1ml of the sealed phase into an immune tube, and incubating for 2 hours (first round and second round) or 1 hour (third round and fourth round) at room temperature; PBST (tween 0.05%) washed 10 times (first, second) or 15 times (third, fourth); adding 800. Mu.l of Gly-HCl buffer solution with pH of 2.2 for elution, and immediately adding 400. Mu.l of Tris-HCl buffer solution with pH of 8.0 for neutralization after 8 minutes; adding into 20ml E.coli SS320 with OD of about 0.8 in logarithmic growth phase, mixing, and standing at 37deg.C for 1 hr; taking out 1ml of bacterial liquid for determining phage titer and glycerol for bacteria preservation; coating the rest bacterial liquid on a flat plate, and culturing overnight at 37 ℃ in an incubator; the next day, the plate was scraped off, and the plates were inoculated into 80ml of 2YT-Amp medium at a certain ratio to have an OD of 0.3, and when the OD reached 0.8 for several hours, 20:1 proportion of helper phase, and then uniformly mixing and standing for 1h at 37 ℃; adding IPTG and Kan antibiotics, shaking and culturing at 250rpm and 30 ℃ overnight; the supernatant was collected, phage was precipitated with PEG/NaCl solution and resuspended in 1.5ml PBS buffer; the resuspended phage was used for the next round of enrichment screening, after 4 rounds of panning, significant enrichment was observed. Phage antibody clones obtained from panning were identified by ELISA: human BCMA protein (Shanghai Proenergy cell medicine technologies Co., ltd.) was coated on 96-well ELISA plates at a concentration of 1. Mu.g/ml overnight at 4 ℃. The non-specific binding sites were then blocked with 5% nonfat milk powder and after extensive washing, the monoclonal phage supernatant was added to a 96-well plate and incubated for 2 hours at 37 ℃. After extensive washing, anti-M13-HRP (GE healthcare, 27-9421-01) was added, reacted for 45min at 37℃and developed by adding TMB after extensive washing, reacted for 5-10min at room temperature, and finally the reaction was stopped with sulfuric acid, and the OD of each well was measured at 450 nm.

The 6 phage antibody clones that specifically bound human BCMA (i.e., the isolated BCMA antibodies described herein) were obtained by ELISA identification, VH and VL gene sequences were obtained after sequencing, and the 6 phage antibody clones were designated 8a7,8a10,8a11, hbma 19, hbma 22, hbma 23, respectively, and the VH and VL gene sequences of the 6 phage antibody clones are shown in table 1.

TABLE 1.6 cloned VH and VL sequences

EXAMPLE 2 expression and purification of anti-BCMA intact antibodies

Phage antibody clones 8a7,8a10,8a11, hbma 19, hbma 22, hbma 23 and positive control antibody chc11d5.3 (patent CN 102421801B) were reconstructed onto eukaryotic expression vectors. Briefly, antibody clones 8a7,8a10,8a11, hbma 19, hbma 22, hbma 23 and chc11d5.3 were redesigned to be intact IgG1, kappa antibodies (i.e., the isolated antigen binding proteins described herein) by: designing a primer to carry out PCR amplification on the VH cloned by the phage antibody, and cloning a PCR product to a pCMV-IgG1NDL vector subjected to AgeI and SalI double enzyme digestion through recombination; primers were designed to PCR amplify VL of phage antibody clones and the PCR products were cloned by recombination into the AgeI and BsiWI double digested pCMV-kappa vector. After the sequencing is correct, the heavy chain expression vector and the light chain expression vector are co-transfected 293F cells to carry out transient expression, and the complete IgG1, kappa antibodies of 8A7,8A10,8A11, hBCMA19, hBCMA22, hBCMA23 and chC11D5.3 are obtained by purifying through a ProteinA column. The gene sequences of the light and heavy chains of 8A7,8a10,8a11, hbma 19, hbma 22, hbma 23 and chc11d5.3 whole antibodies are shown in table 2.

TABLE 2 light and heavy chain sequences of intact antibodies

Example 3 detection of binding affinity of anti-BCMA intact antibodies

The binding affinity of 8A7, 8a10, 8a11, hbma 19, hbma 22, hbma 23 and chc11d5.3 whole antibodies obtained in example 2 against recombinant human BCMA protein was measured using an Octet RED384 instrument (Pall ForteBio). First, human BCMA protein was labeled with Biotin (EZ-Link Sulfo-NHS-LC-Biotin, pierce, 21327). Binding kinetics analysis between antigen and antibody was performed by biofilm interference (BLI) technique using a molecular interaction analyzer fortebioctetred 384 (sall) (PBS buffer with 0.1% bsa and 0.02% tween 20 for both antigen and antibody dilutions). Immobilized with SA sensor using biotin-conjugated antigen at a concentration of 20nM, 1500rpm/min, 2min; then bound to a double diluted antibody (i.e. 8A7, 8A10, 8A11, hBCMA19, hBCMA22, hBCMA23 and chC11D5.3 whole antibody) solution obtained in example 2 for 5min at 1500rpm/min. Finally, dissociation was carried out for 20 minutes at 1500rpm/min. The remaining antibody was regenerated by glycine pulsing. The obtained results were subjected to data analysis by Octet Data Analysis 9.0.9.0 software (fortebio) to calculate the binding strength of antigen and antibody to obtain K _D Values, ka (1/Ms) values and Kd (1/s) values. The intact antibodies 8A7, 8a10,8a11, hbma 19, hbma 22, hbma 23, chc11d5.3 against BCMA all bind with high affinity to recombinant human BCMA protein (as shown in table 3).

TABLE 3 binding affinity of anti-BCMA intact antibodies to human BCMA protein

In table 3, ka: a binding rate constant; kd: dissociation rate constant; k (K) _D : affinity constant, equal to Kd/Ka.

As can be seen from Table 3, the dissociation rates of 8A7, 8A10,8A11, hBCMA19, hBCMA22, hBCMA23 complete antibodies were smaller than that of the control antibody chC11D5.3, so that the affinities of 8A7, 8A10,8A11, hBCMA19, hBCMA22, hBCMA23 complete antibodies were higher than that of the control antibody chC11D5.3.

EXAMPLE 4 detection of anti-BCMA intact antibody and cell surface BCMA binding Activity

Cell surface target antigen (BCMA) binding activity assays were performed with antibodies (i.e., 8A7, 8a10,8a11, hbma 19, hbma 22, hbma 23, and chc11d5.3 whole antibodies obtained in example 2) using an iQue screen flow machine (purchased from intelllicyt corporation) using PBS containing 0.1% bsa as buffer by flow cytometry (FACS):

1. concentration was 1X10 using buffer ⁶ Target cells (i.e., MM1S myeloma cells) at cells/ml, 96 Kong Jian bottom plates (burning 3894) were added, 30. Mu.l per well;

2. The concentration of the detection antibody is 20 mug/ml by using buffer solution, and the antibody is diluted according to the ratio of 3 times, so as to form 8 concentration gradients;

3. adding the prepared antibodies with different concentrations into paved target cells according to 30 μl/hole, and uniformly mixing;

incubating for 1 hour at 4.4 ℃ in a refrigerator;

5. 150 μl of buffer is added to each well, 300g is centrifuged for 5 minutes, and the cells are loosened after the supernatant is discarded;

6. repeating the step 5;

7. preparing a fluorescent secondary antibody (ab 98593) by using a buffer solution according to a ratio of 1:200, adding 30 mu l of each hole into cells, uniformly mixing, and incubating for 30 minutes at 4 ℃ in a refrigerator;

8. 150 μl of buffer is added to each well, 300g is centrifuged for 5 minutes, and the cells are loosened after the supernatant is discarded;

9. repeating the step 8;

10. after adding 35. Mu.l of buffer to each well, the mixture was measured by a flow meter.

The analytical results of the streaming affinity binding experiments are shown in table 4 and fig. 1.

TABLE 4 binding results of anti-BCMA intact antibodies to cell surface BCMA proteins

Intact antibodies	8A7	8A10	8A11	hBCMA19	hBCMA22	hBCMA23	chC11D5.3
								EC50(μg/ml)	1.31	0.50	1.25	0.33	0.68	0.71	1.55

As can be seen from table 4, 8A7, 8a10,8a11, hbma a19, hbma a22, hbma a23 complete antibodies have stronger binding activity to myeloma cell MM1S that low-expressed BCMA antigen; and are all lower than the EC50 value of the positive control antibody chc11d5.3. As can be seen from FIG. 1, the average fluorescence intensities of the whole antibodies of 8A7, 8A10,8A11, hBCMA19, hBCMA22 and hBCMA23 binding to MMIS of myeloma cells are high.

Example 5 ADCC Activity assay of anti-BCMA intact antibody

ADCC Activity of anti-BCMA intact antibodies (i.e., 8A7,8A10, 8A11, hBCMA19, hBCMA22, hBCMA23 and chC11D5.3 intact antibodies) against BCMA antigen-expressed myeloma cell MM1S was measured by CytotoxThe nonradioactive cytotoxicity assay kit (Promega) was used for the assay. The specific treatment process of the cell and antibody sample is as follows:

resuspension of MM1S cells was collected with X-VIVO (LONZA) medium containing 5% FBS (GIBICO) to form a total of 2.0x10 ⁵ cell suspensions of cells/ml were used. Take 8x10 ⁷ PBMC cells (of healthy humans) were resuspended to 16ml with X-VIVO (LONZA) medium containing 5% FBS (GIBICO) to form 5.0x10 ⁶ cell suspensions of cells/ml were used. The 8A7,8a10, 8a11, hbma 19, hbma 22, hbma 23 and positive control chc11d5.3 whole antibodies were diluted with medium to different antibody concentrations. Next, after plating (96-well V-plate (Thermo)) 50. Mu.l of MM1S cell suspension, 100. Mu.l of PBMC cell suspension and 50. Mu.l of each antibody solution per well, CO at 37℃was used ₂ Incubate in incubator for 24 hours. Mu.l of MM1S cell suspension was added to 100. Mu.l of PBMC cell suspension as a control well. According to CytotoxThe ADCC activity of the antibody is detected by a non-radioactive cytotoxicity detection kit (Promega) instruction, and the calculation formula is as follows: cytotoxicity% = 100X (experiment Kong control wells)/(maximum killing of target cells-spontaneous release of target cells). The experimental results are shown in FIG. 2, wherein isotype control refers to intact IgG1, kappa antibodies that are not associated with this target. As can be seen from fig. 2, 8A7,8A10, 8a11, hbma 19, hbma 22, hbma 23 complete antibodies were able to effectively mediate efficient killing of BCMA positive myeloma cells MM1S, and had better killing effect than positive control chc11d5.3, i.e. more remarkable ADCC activity than positive control.

Example 6 CDC Activity detection of anti-BCMA intact antibody

The ability of anti-BCMA complete antibodies to elicit CDC effects against CHO-BCMA cells was tested using a cytotoxicity test kit (Promega, cat#g1780) as follows:

1. preparing a culture medium (A: DMEM+2% FBS, B: DMEM+2% FBS+10% rabbit complement);

2. target cells CHO-BCMA were centrifuged at 400g for 5 min and then the cells were resuspended in medium A as described above to a target cell density of 4X10 ⁵ Individual cells/ml, 100 ul/well were added to 96 well cell culture plates;

3. the antibodies were diluted with medium B as described above, 10ug/ml initial concentration, 3-fold gradient dilution, and 8 gradients total. Adding 100 ul/hole into a detection hole, wherein two compound holes are arranged at each concentration point, and chC11D5.3 is used as a positive control antibody;

4. complement was Rabbit complement Rabbit compact 3-4week (Cat # 31061-3) from PelFreez Bio company, and the final concentration of the experiment used for complement was 5%;

5. control wells were set as required for the kit, and after incubation of the obtained mixture in an incubator at 37 ℃ for 4 hours, the absorbance of 490nM was recorded by the microplate reader, and the percent of target cell lysis was calculated using the formula given by the kit, graphpad prism 8 analysis data.

The experimental results are shown in FIG. 3, wherein isotype control refers to intact IgG1, kappa antibodies that are not associated with this target. As can be seen from fig. 3, both 8a10, hbma 22 and hbma 23 complete antibodies induced strong CDC effects in a dose-dependent manner, i.e. all had significant CDC activity, with 8a10 complete antibodies killing better than the positive control chc11d5.3.

Example 7 anti-BCMA intact antibody reduces tumor burden in xenograft mice

We injected 6x 10 in tail vein using NOD scid common gamma chain knockout (NSG) mice lacking functional B, T and NK cell populations ⁶ After MM1S-luc cells, mice were randomized into 3 groups of 8. The first group received an equivalent dose of PBS until the end of the experiment, while the second and third groups received 200ug injections twice a week of the experimental group antibody and isotype control antibody, respectively. Intraperitoneal (i.p.) administration of antibody for 4 weeks starting on the day of tumor cell injectionTime. Tumor growth in mice was monitored weekly using a mouse in vivo imaging system and bioluminescence was measured 3 minutes after intraperitoneal injection of fluorescein.

The experimental results are shown in fig. 4, starting from the first measurement point on day 7, the group treated with hbma 22 antibody showed significantly lower tumor burden. In addition, the group showed less overall tumor burden during the entire monitoring period. On day 28 after target cell injection, as shown in fig. 5, a higher tumor burden was seen in mice receiving isotype control antibodies. Mice receiving BCMA antibodies were able to significantly inhibit tumor growth.

The foregoing detailed description is provided by way of explanation and example and is not intended to limit the scope of the appended claims. Numerous variations of the presently exemplified embodiments of the present application will be apparent to those of ordinary skill in the art and remain within the scope of the appended claims and equivalents thereof.

Sequence listing

<110> Shanghai Primary energy cytomedical technology Co., ltd

<120> an isolated antigen BCMA-binding protein and uses thereof

<130> 0065-PA-029

<160> 65

<170> PatentIn version 3.5

<210> 1

<211> 15

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7 LCDR1

<400> 1

Arg Ala Ser Glu Ser Val Ser Val Ser Gly Ile His Leu Leu His

1 5 10 15

<210> 2

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/hBCMA19/ LCDR2

<400> 2

Leu Gly Arg Asn Arg Ala Ala

1 5

<210> 3

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/ hBCMA23 LCDR3

<400> 3

Leu Gln Ser Arg Arg Phe Pro Arg His

1 5

<210> 4

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 HCDR1

<400> 4

Asp Tyr Thr Ile Asn

1 5

<210> 5

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A11 HCDR2

<400> 5

Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe Arg

1 5 10 15

Gly

<210> 6

<211> 8

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 HCDR3

<400> 6

Asp Ile Arg Tyr Val Met Asp Tyr

1 5

<210> 7

<211> 15

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A10/8A11/hBCMA19/hBCMA22/hBCMA23 LCDR1

<400> 7

Arg Ala Ser Glu Ser Val Ser Val Pro Gly Ile Ser Phe Ile His

1 5 10 15

<210> 8

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A10/hBCMA23 HCDR2

<400> 8

Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe Arg

1 5 10 15

Ser

<210> 9

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A11/hBCMA22/hBCMA23 LCDR2

<400> 9

Leu Gly Ser Asn Arg Pro Ser

1 5

<210> 10

<211> 15

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> LCDR1 formula

<220>

<221> misc_feature

<222> (9)..(9)

<223> Xaa=Pro or Ser

<220>

<221> misc_feature

<222> (12)..(12)

<223> Xaa=His or Ser

<220>

<221> misc_feature

<222> (13)..(13)

<223> Xaa=Phe or Leu

<220>

<221> misc_feature

<222> (14)..(14)

<223> Xaa=lie or Leu

<400> 10

Arg Ala Ser Glu Ser Val Ser Val Xaa Gly Ile Xaa Xaa Xaa His

1 5 10 15

<210> 11

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> LCDR2 general formula

<220>

<221> misc_feature

<222> (3)..(3)

<223> Xaa=Arg or Ser

<220>

<221> misc_feature

<222> (6)..(6)

<223> Xaa=Ala or Pro

<220>

<221> misc_feature

<222> (7)..(7)

<223> Xaa=Ala or Ser

<400> 11

Leu Gly Xaa Asn Arg Xaa Xaa

1 5

<210> 12

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11 HCDR2 general formula

<220>

<221> misc_feature

<222> (17)..(17)

<223> Xaa=Gly or Ser

<400> 12

Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe Arg

1 5 10 15

Xaa

<210> 13

<211> 23

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 L-FR1

<400> 13

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys

20

<210> 14

<211> 15

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 L-FR2

<400> 14

Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr

1 5 10 15

<210> 15

<211> 32

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 L-FR3

<400> 15

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys

20 25 30

<210> 16

<211> 10

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 L-FR4

<400> 16

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 17

<211> 30

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11 H-FR1

<400> 17

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala

20 25 30

<210> 18

<211> 14

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10 H-FR2

<400> 18

Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly

1 5 10

<210> 19

<211> 32

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11 H-FR3

<400> 19

Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr Leu Gln

1 5 10 15

Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

20 25 30

<210> 20

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 H-FR4

<400> 20

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 21

<211> 14

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A11 H-FR2

<400> 21

Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly

1 5 10

<210> 22

<211> 14

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A 11H-FR 2 general formula

<220>

<221> misc_feature

<222> (13)..(13)

<223> Xaa=lie or Met

<400> 22

Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Xaa Gly

1 5 10

<210> 23

<211> 111

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7 VL

<400> 23

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Val Ser

20 25 30

Gly Ile His Leu Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Gly Arg Asn Arg Ala Ala Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Arg Phe Pro Arg His Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 24

<211> 111

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A10/hBCMA19 VL

<400> 24

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Val Pro

20 25 30

Gly Ile Ser Phe Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Gly Arg Asn Arg Ala Ala Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Arg Phe Pro Arg His Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 25

<211> 111

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A11/hBCMA22/hBCMA23 VL

<400> 25

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Val Pro

20 25 30

Gly Ile Ser Phe Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Gly Ser Asn Arg Pro Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Arg Phe Pro Arg His Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 26

<211> 111

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> VL general formula

<220>

<221> misc_feature

<222> (32)..(32)

<223> Xaa=Pro or Ser

<220>

<221> misc_feature

<222> (35)..(35)

<223> Xaa=His or Ser

<220>

<221> misc_feature

<222> (36)..(36)

<223> Xaa=Phe or Leu

<220>

<221> misc_feature

<222> (37)..(37)

<223> Xaa=lie or Leu

<220>

<221> misc_feature

<222> (56)..(56)

<223> Xaa=Arg or Ser

<220>

<221> misc_feature

<222> (59)..(59)

<223> Xaa=Ala or Pro

<220>

<221> misc_feature

<222> (60)..(60)

<223> Xaa=Ala or Ser

<400> 26

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Val Xaa

20 25 30

Gly Ile Xaa Xaa Xaa His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Gly Xaa Asn Arg Xaa Xaa Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Arg Phe Pro Arg His Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 27

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7 VH

<400> 27

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 28

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A10 VH

<400> 28

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Ser Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 29

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A11 VH

<400> 29

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 30

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11 VH general formula

<220>

<221> misc_feature

<222> (48)..(48)

<223> Xaa=lie or Met

<220>

<221> misc_feature

<222> (66)..(66)

<223> Xaa=Gly or Ser

<400> 30

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Xaa

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Xaa Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 31

<211> 107

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 light chain constant region

<400> 31

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 32

<211> 330

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7/8A10/8A11/hBCMA19/hBCMA22/hBCMA23 heavy chain constant region

<400> 32

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 33

<211> 218

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7 light chain

<400> 33

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Val Ser

20 25 30

Gly Ile His Leu Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Gly Arg Asn Arg Ala Ala Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Arg Phe Pro Arg His Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 34

<211> 218

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A10/hBCMA19 light chain

<400> 34

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Val Pro

20 25 30

Gly Ile Ser Phe Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Gly Arg Asn Arg Ala Ala Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Arg Phe Pro Arg His Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 35

<211> 218

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A11/hBCMA22/hBCMA23 light chain

<400> 35

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Val Pro

20 25 30

Gly Ile Ser Phe Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Gly Ser Asn Arg Pro Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Ile Tyr Tyr Cys Leu Gln Ser Arg

85 90 95

Arg Phe Pro Arg His Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 36

<211> 447

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A7 heavy chain

<400> 36

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 37

<211> 447

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A10 heavy chain

<400> 37

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Ser Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 38

<211> 447

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> 8A11 heavy chain

<400> 38

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Val Lys Gln Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 39

<211> 333

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A7 VL

<400> 39

gaaatcgttc tgacccagag tccagcgacg ctgagtctga gtccgggtga acgcgcgacg 60

ctgagttgcc gtgccagcga aagcgttagc gtgagtggca ttcacttact ccactggtac 120

cagcagaaac cgggccaagc cccacgtctg ctgatctatc tgggacgcaa tcgggcggcc 180

ggcattccag cccgctttag cggcagcggt agcggtaccg attttacgct gaccatcagc 240

agtctggagc cagaagatgc cgccatctac tactgtttac agagcagaag gtttcctcga 300

cacttcggcc aaggtaccaa actggaaatc aag 333

<210> 40

<211> 333

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> code 8A10/hBCMA19 VL

<400> 40

gaaatcgttc tgacccagag tccagcgacg ctgagtctga gtccgggtga acgcgcgacg 60

ctgagttgcc gtgccagcga aagcgttagc gtgcctggca tatctttcat ccactggtac 120

cagcagaaac cgggccaagc cccacgtctg ctgatctatc tgggacgcaa tcgggcggcc 180

ggcattccag cccgctttag cggcagcggt agcggtaccg attttacgct gaccatcagc 240

agtctggagc cagaagatgc cgccatctac tactgtttac agagcagaag gtttcctcga 300

cacttcggcc aaggtaccaa actggaaatc aag 333

<210> 41

<211> 333

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A11/hBCMA22/hBCMA23 VL

<400> 41

gaaatcgttc tgacccagag tccagcgacg ctgagtctga gtccgggtga acgcgcgacg 60

ctgagttgcc gtgccagcga aagcgttagc gtgcctggca tatctttcat ccactggtac 120

cagcagaaac cgggccaagc cccacgtctg ctgatctatc tgggcagcaa tcgaccatcc 180

ggcattccag cccgctttag cggcagcggt agcggtaccg attttacgct gaccatcagc 240

agtctggagc cagaagatgc cgccatctac tactgtttac agagcagaag gtttcctcga 300

cacttcggcc aaggcaccaa actggaaatc aag 333

<210> 42

<211> 351

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A7 VH

<400> 42

caagttcagc tcgtgcagag cggctccgag gtgaagcagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata cagcttcgcc gattacacca tcaactgggt tcgtcaagct 120

cccggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaggtcg tttcgtgttc tctttagaca caagcgcgag caccgcctat 240

ttacagatca gctctttaaa ggccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag c 351

<210> 43

<211> 351

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A10 VH

<400> 43

caagttcagc tcgtgcagag cggctccgag gtgaagcagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata cagcttcgcc gattacacca tcaactgggt tcgtcaagct 120

cccggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaagtcg tttcgtgttc tctttagaca caagcgcgag caccgcctat 240

ttacagatca gctctttaaa ggccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag c 351

<210> 44

<211> 351

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A11 VH

<400> 44

caagttcagc tcgtgcagag cggctccgag gtgaagcagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata cagcttcgcc gattacacca tcaactgggt tcgtcaagct 120

cccggtcaag gtctggagtg gataggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaggtcg tttcgtgttc tctttagaca caagcgcgag caccgcctat 240

ttacagatca gctctttaaa ggccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag c 351

<210> 45

<211> 654

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> coding 8A7 light chain

<400> 45

gaaatcgttc tgacccagag tccagcgacg ctgagtctga gtccgggtga acgcgcgacg 60

ctgagttgcc gtgccagcga aagcgttagc gtgagtggca ttcacttact ccactggtac 120

cagcagaaac cgggccaagc cccacgtctg ctgatctatc tgggacgcaa tcgggcggcc 180

ggcattccag cccgctttag cggcagcggt agcggtaccg attttacgct gaccatcagc 240

agtctggagc cagaagatgc cgccatctac tactgtttac agagcagaag gtttcctcga 300

cacttcggcc aaggtaccaa actggaaatc aagcgtacgg tggctgcacc atctgtcttc 360

atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg 420

aataacttct accccagaga agccaaagtg cagtggaagg tggacaacgc cctgcagagc 480

ggaaacagcc aggaaagcgt gacagagcag gattccaagg attccacata cagcctgagc 540

agcacactga cactgtccaa ggccgactac gagaagcaca aggtgtacgc ctgcgaagtg 600

acacaccagg gactgtcctc ccctgtgaca aagagcttca acagaggaga atgc 654

<210> 46

<211> 654

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A10/hBCMA19 light chain

<400> 46

gaaatcgttc tgacccagag tccagcgacg ctgagtctga gtccgggtga acgcgcgacg 60

ctgagttgcc gtgccagcga aagcgttagc gtgcctggca tatctttcat ccactggtac 120

cagcagaaac cgggccaagc cccacgtctg ctgatctatc tgggacgcaa tcgggcggcc 180

ggcattccag cccgctttag cggcagcggt agcggtaccg attttacgct gaccatcagc 240

agtctggagc cagaagatgc cgccatctac tactgtttac agagcagaag gtttcctcga 300

cacttcggcc aaggtaccaa actggaaatc aagcgtacgg tggctgcacc atctgtcttc 360

atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg 420

aataacttct accccagaga agccaaagtg cagtggaagg tggacaacgc cctgcagagc 480

ggaaacagcc aggaaagcgt gacagagcag gattccaagg attccacata cagcctgagc 540

agcacactga cactgtccaa ggccgactac gagaagcaca aggtgtacgc ctgcgaagtg 600

acacaccagg gactgtcctc ccctgtgaca aagagcttca acagaggaga atgc 654

<210> 47

<211> 654

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A11/hBCMA22/hBCMA23 light chain

<400> 47

gaaatcgttc tgacccagag tccagcgacg ctgagtctga gtccgggtga acgcgcgacg 60

ctgagttgcc gtgccagcga aagcgttagc gtgcctggca tatctttcat ccactggtac 120

cagcagaaac cgggccaagc cccacgtctg ctgatctatc tgggcagcaa tcgaccatcc 180

ggcattccag cccgctttag cggcagcggt agcggtaccg attttacgct gaccatcagc 240

agtctggagc cagaagatgc cgccatctac tactgtttac agagcagaag gtttcctcga 300

cacttcggcc aaggcaccaa actggaaatc aagcgtacgg tggctgcacc atctgtcttc 360

atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg 420

aataacttct accccagaga agccaaagtg cagtggaagg tggacaacgc cctgcagagc 480

ggaaacagcc aggaaagcgt gacagagcag gattccaagg attccacata cagcctgagc 540

agcacactga cactgtccaa ggccgactac gagaagcaca aggtgtacgc ctgcgaagtg 600

acacaccagg gactgtcctc ccctgtgaca aagagcttca acagaggaga atgc 654

<210> 48

<211> 1341

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> coding 8A7 heavy chain

<400> 48

caagttcagc tcgtgcagag cggctccgag gtgaagcagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata cagcttcgcc gattacacca tcaactgggt tcgtcaagct 120

cccggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaggtcg tttcgtgttc tctttagaca caagcgcgag caccgcctat 240

ttacagatca gctctttaaa ggccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag cgcgtcgacc 360

aagggcccat cggtcttccc cctggcaccc tcctccaaga gcacctctgg gggcacagcg 420

gccctgggct gcctggtcaa ggactacttc cccgaacctg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacccagac ctacatctgc 600

aacgtgaatc acaagcccag caacaccaag gtggacaaga aagttgagcc caaatcttgt 660

gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 720

ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 780

tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 840

ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 900

cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 960

tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 1020

gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 1080

aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 1140

tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 1200

gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg 1260

aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 1320

ctctccctgt ctccgggtaa a 1341

<210> 49

<211> 1341

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A10 heavy chain

<400> 49

caagttcagc tcgtgcagag cggctccgag gtgaagcagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata cagcttcgcc gattacacca tcaactgggt tcgtcaagct 120

cccggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaagtcg tttcgtgttc tctttagaca caagcgcgag caccgcctat 240

ttacagatca gctctttaaa ggccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag cgcgtcgacc 360

aagggcccat cggtcttccc cctggcaccc tcctccaaga gcacctctgg gggcacagcg 420

gccctgggct gcctggtcaa ggactacttc cccgaacctg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacccagac ctacatctgc 600

aacgtgaatc acaagcccag caacaccaag gtggacaaga aagttgagcc caaatcttgt 660

gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 720

ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 780

tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 840

ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 900

cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 960

tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 1020

gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 1080

aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 1140

tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 1200

gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg 1260

aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 1320

ctctccctgt ctccgggtaa a 1341

<210> 50

<211> 1341

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding 8A11 heavy chain

<400> 50

caagttcagc tcgtgcagag cggctccgag gtgaagcagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata cagcttcgcc gattacacca tcaactgggt tcgtcaagct 120

cccggtcaag gtctggagtg gataggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaggtcg tttcgtgttc tctttagaca caagcgcgag caccgcctat 240

ttacagatca gctctttaaa ggccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag cgcgtcgacc 360

aagggcccat cggtcttccc cctggcaccc tcctccaaga gcacctctgg gggcacagcg 420

gccctgggct gcctggtcaa ggactacttc cccgaacctg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacccagac ctacatctgc 600

aacgtgaatc acaagcccag caacaccaag gtggacaaga aagttgagcc caaatcttgt 660

gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 720

ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 780

tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 840

ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 900

cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 960

tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 1020

gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 1080

aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 1140

tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 1200

gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg 1260

aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 1320

ctctccctgt ctccgggtaa a 1341

<210> 51

<211> 231

<212> PRT

<213> human (homosapiens)

<400> 51

Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

1 5 10 15

Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

20 25 30

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

35 40 45

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

50 55 60

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

65 70 75 80

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

85 90 95

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

100 105 110

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

115 120 125

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys

130 135 140

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

145 150 155 160

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

165 170 175

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

180 185 190

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

195 200 205

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

210 215 220

Leu Ser Leu Ser Pro Gly Lys

225 230

<210> 52

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22 HCDR2

<400> 52

Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp Phe Arg

1 5 10 15

Gly

<210> 53

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22/hBCMA23 HCDR2 general formula

<220>

<221> misc_feature

<222> (10)..(10)

<223> Xaa=Ala or Thr

<220>

<221> misc_feature

<222> (17)..(17)

<223> Xaa=Gly or Ser

<400> 53

Trp Ile Asn Thr Glu Thr Arg Glu Pro Xaa Tyr Ala Tyr Asp Phe Arg

1 5 10 15

Xaa

<210> 54

<211> 30

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22/hBCMA23 H-FR1

<400> 54

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala

20 25 30

<210> 55

<211> 14

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22/hBCMA23 H-FR2

<400> 55

Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met Gly

1 5 10

<210> 56

<211> 32

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22/hBCMA23 H-FR3

<400> 56

Arg Phe Thr Phe Thr Leu Asn Thr Ser Ile Ser Thr Ala Tyr Leu Glu

1 5 10 15

Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

20 25 30

<210> 57

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22 VH

<400> 57

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Thr Phe Thr Leu Asn Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 58

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA23 VH

<400> 58

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Ser Arg Phe Thr Phe Thr Leu Asn Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 59

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22/hBCMA23VH formula

<220>

<221> misc_feature

<222> (59)..(59)

<223> Xaa=Ala or Thr

<220>

<221> misc_feature

<222> (66)..(66)

<223> Xaa=Gly or Ser

<400> 59

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Xaa Tyr Ala Tyr Asp Phe

50 55 60

Arg Xaa Arg Phe Thr Phe Thr Leu Asn Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 60

<211> 447

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA19/hBCMA22 heavy chain

<400> 60

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Ala Tyr Ala Tyr Asp Phe

50 55 60

Arg Gly Arg Phe Thr Phe Thr Leu Asn Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 61

<211> 447

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> hBCMA23 heavy chain

<400> 61

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Asp Tyr

20 25 30

Thr Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Arg Glu Pro Thr Tyr Ala Tyr Asp Phe

50 55 60

Arg Ser Arg Phe Thr Phe Thr Leu Asn Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ile Arg Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 62

<211> 351

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding hBCMA19/hBCMA22 VH

<400> 62

caagttcagc tcgtgcagag cggcgccgag gtgaagaagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata caccttcgcc gattacacca tcaactgggt tcgtcaagct 120

accggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccgcctat 180

gcctacgact tcagaggtcg tttcacgttc actttaaaca caagcatcag caccgcctat 240

ttagagctca gctctttaag gtccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag c 351

<210> 63

<211> 351

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding hBCMA23 VH

<400> 63

caagttcagc tcgtgcagag cggcgccgag gtgaagaagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata caccttcgcc gattacacca tcaactgggt tcgtcaagct 120

accggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaagtcg tttcacgttc actttaaaca caagcatcag caccgcctat 240

ttagagctca gctctttaag gtccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag c 351

<210> 64

<211> 1341

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding hBCMA19/hBCMA22 heavy chain

<400> 64

caagttcagc tcgtgcagag cggcgccgag gtgaagaagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata caccttcgcc gattacacca tcaactgggt tcgtcaagct 120

accggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccgcctat 180

gcctacgact tcagaggtcg tttcacgttc actttaaaca caagcatcag caccgcctat 240

ttagagctca gctctttaag gtccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag cgcgtcgacc 360

aagggcccat cggtcttccc cctggcaccc tcctccaaga gcacctctgg gggcacagcg 420

gccctgggct gcctggtcaa ggactacttc cccgaacctg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacccagac ctacatctgc 600

aacgtgaatc acaagcccag caacaccaag gtggacaaga aagttgagcc caaatcttgt 660

gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 720

ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 780

tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 840

ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 900

cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 960

tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 1020

gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 1080

aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 1140

tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 1200

gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg 1260

aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 1320

ctctccctgt ctccgggtaa a 1341

<210> 65

<211> 1341

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<220>

<223> encoding hBCMA23 heavy chain

<400> 65

caagttcagc tcgtgcagag cggcgccgag gtgaagaagc ccggcgcttc cgtgaaggtg 60

agctgcaagg ccagcggata caccttcgcc gattacacca tcaactgggt tcgtcaagct 120

accggtcaag gtctggagtg gatgggctgg atcaacaccg aaaccagaga gcccacctat 180

gcctacgact tcagaagtcg tttcacgttc actttaaaca caagcatcag caccgcctat 240

ttagagctca gctctttaag gtccgaggat accgccgtgt actactgcgc tcgtgacatc 300

agatacgtga tggactactg gggccaaggt acactggtga ccgtgtccag cgcgtcgacc 360

aagggcccat cggtcttccc cctggcaccc tcctccaaga gcacctctgg gggcacagcg 420

gccctgggct gcctggtcaa ggactacttc cccgaacctg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacccagac ctacatctgc 600

aacgtgaatc acaagcccag caacaccaag gtggacaaga aagttgagcc caaatcttgt 660

gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 720

ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 780

tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 840

ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 900

cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 960

tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 1020

gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 1080

aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 1140

tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 1200

gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg 1260

aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 1320

ctctccctgt ctccgggtaa a 1341

Claims

1. An antibody or antigen-binding fragment thereof capable of binding BCMA, said antibody or antigen-binding fragment thereof comprising a heavy chain variable region VH comprising HCDR1, HCDR2 and HCDR3 and a light chain variable region VL comprising

LCDR1, LCDR2 and LCDR3, said HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3

The amino acid sequence of LCDR3 is selected from any one of the following groups:

(1) The amino acid sequence of the HCDR1 is shown as SEQ ID NO. 4, the amino acid sequence of the HCDR2 is shown as SEQ ID NO. 52, the amino acid sequence of the HCDR3 is shown as SEQ ID NO. 6, and the amino acid sequence of the HCDR2 is shown as SEQ ID NO. 52

The amino acid sequence of LCDR1 is shown as SEQ ID NO. 7, the amino acid sequence of LCDR2 is shown as SEQ ID NO. 2, and the amino acid sequence of LCDR3 is shown as SEQ ID NO. 3; and

(2) The amino acid sequence of the HCDR1 is shown as SEQ ID NO. 4, the amino acid sequence of the HCDR2 is shown as SEQ ID NO. 52, the amino acid sequence of the HCDR3 is shown as SEQ ID NO. 6, and the amino acid sequence of the HCDR2 is shown as SEQ ID NO. 52

The amino acid sequence of LCDR1 is shown as SEQ ID NO. 7, the amino acid sequence of LCDR2 is shown as SEQ ID NO. 9, and the amino acid sequence of LCDR3 is shown as SEQ ID NO. 3.

2. The antibody or antigen-binding fragment thereof of claim 1, wherein the antigen-binding fragment comprises Fab, fab ', F (ab) 2, fv fragment, F (ab') 2, scFv, and/or di-scFv.

3. The antibody or antigen-binding fragment thereof of claim 1, wherein the VL comprises the framework regions L-FR1, L-

FR2, L-FR3, and L-FR4, wherein the amino acid sequence of said L-FR1 is shown as SEQ ID NO. 13, the amino acid sequence of said L-FR2 is shown as SEQ ID NO. 14, the amino acid sequence of said L-FR3 is shown as SEQ ID NO. 15, and the amino acid sequence of said L-FR4 is shown as SEQ ID NO. 16.

4. The antibody or antigen-binding fragment thereof of claim 1, wherein the VH comprises framework regions H-FR1, H-

FR2, H-FR3, and H-FR4, the amino acid sequence of said H-FR1 being shown in SEQ ID NO:54, said H-

The amino acid sequence of FR2 is shown as SEQ ID NO. 55, the amino acid sequence of H-FR3 is shown as SEQ ID NO. 56, and the amino acid sequence of H-FR4 is shown as SEQ ID NO. 20.

5. The antibody or antigen-binding fragment thereof of claim 1, wherein the amino acid sequences of VH and VL are selected from any one of the following groups:

(1) The amino acid sequence of the VH is shown as SEQ ID NO. 57, and the amino acid sequence of the VL is shown as SEQ ID NO. 24; and

(2) The amino acid sequence of the VH is shown as SEQ ID NO. 57, and the amino acid sequence of the VL is shown as SEQ ID NO. 25.

6. The antibody or antigen binding fragment thereof of claim 1, comprising an antibody light chain constant region, and the antibody light chain constant region is a human igκ constant region.

7. The antibody or antigen-binding fragment thereof of claim 6, wherein the amino acid sequence of the antibody light chain constant region is set forth in SEQ ID No. 31.

8. The antibody or antigen-binding fragment thereof of claim 1, comprising an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgG heavy chain constant region.

9. The antibody or antigen-binding fragment thereof of claim 1, comprising an antibody heavy chain constant region, and the antibody heavy chain constant region is derived from a human IgG1 heavy chain constant region.

10. The antibody or antigen-binding fragment thereof of claim 8, wherein the amino acid sequence of the antibody heavy chain constant region is set forth in SEQ ID No. 32.

11. The antibody or antigen binding fragment thereof of claim 1, comprising an antibody heavy chain HC and an antibody light chain LC, and the amino acid sequences of HC and LC are selected from any one of the following groups:

(1) The amino acid sequence of HC is shown as SEQ ID NO. 60, and the amino acid sequence of LC is shown as SEQ ID NO. 34; and

(2) The amino acid sequence of HC is shown as SEQ ID NO. 60, and the amino acid sequence of LC is shown as SEQ ID NO. 35.

12. An isolated one or more nucleic acid molecules encoding the antibody or antigen-binding fragment thereof of any one of claims 1-11.

13. A vector comprising the nucleic acid molecule of claim 12.

14. A cell comprising the nucleic acid molecule of claim 12 or the vector of claim 13.

15. A method of making the antibody or antigen-binding fragment thereof of any one of claims 1-11, comprising culturing claim 14 under conditions such that the antibody or antigen-binding fragment thereof of any one of claims 1-11 is expressed

The cells.

16. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1-11, the nucleic acid molecule of claim 12, the vector of claim 13 and/or the cell of claim 14, and optionally a pharmaceutically acceptable adjuvant.

17. Use of the antibody or antigen binding fragment thereof of any one of claims 1-11, the nucleic acid molecule of claim 12, the vector of claim 13, the cell of claim 14 and/or the pharmaceutical composition of claim 16 in the manufacture of a medicament for alleviating and/or treating a tumor, which is myeloma.

18. A method of detecting BCMA in a sample, said method comprising administering the antibody or antigen binding fragment thereof according to any one of claims 1-11, said method being a method for the purpose of diagnosis of a non-disease.