Anti-human CD38 antibody and its application

Abstract

The present invention provides an antibody molecule or a fragment thereof that binds to human CD38, and the antibody molecule or the fragment thereof is used for preventing or treating a disease. The antibody molecule of the present invention is a human CD38 antibody molecule having a completely new sequence obtained by screening after immunizing mice and fusing with myeloma cells to produce hybridoma cells, and the antibody has the same or better functional activity as known antibodies, including in vitro ADCC, CDC and Daudi cell apoptosis activity, etc.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to a novel anti-CD38 antibody or a functional fragment thereof and also relates to the application of the antibody or the functional fragment thereof. Background Art

[0002] CD38 is a type II transmembrane glycoprotein with a molecular weight of 46KD, including the C-terminal extracellular region (258 amino acids), the transmembrane region (21 amino acids) and the N-terminus (21 amino acids). Its functions include receptor-mediated adhesion and signal transduction, as well as calcium mobilization through its extracellular enzyme activity, catalyzing the formation of cyclic ADP ribose (cADPR) and ADPR.

[0003] CD38 is uniformly highly expressed on multiple myeloma cells (MM), but is lowly expressed on normal lymphocytes and bone marrow cells and some tissues of non-hematopoietic origin. Since CD38 is a transmembrane glycoprotein with in vitro enzyme activity and also functions as a receptor and adhesion molecule, it is an ideal target for MM treatment; based on its unique mechanism of action, low toxicity and single-agent activity, CD38 antibodies are also suitable for combination regimens. Currently, multiple studies are exploring the efficacy of CD38 antibody-based therapies in newly diagnosed high-risk MM patients. In addition, the therapeutic effect of CD38 antibodies for other hematological malignancies (acute lymphocytic leukemia, NK / T cell lymphoma and acute myeloid leukemia) is also under study.

[0004] Currently, many pharmaceutical companies are developing monoclonal antibodies against CD38, which achieve anti-MM mechanisms through antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), antibody-dependent cellular phagocytosis (ADPC) and direct inhibition of CD38 enzymatic activity. Currently, CD38 antibodies in clinical trials or approved include Daratumumab (Darzalex), Isatuximab, MOR-202 and TAK-079.

[0005] Among the monoclonal antibody drugs currently developed for CD38, Johnson & Johnson's Darzalex is the first CD38-mediated, cytolytic antibody drug approved worldwide. It has a broad-spectrum killing activity and can target and bind to the transmembrane extracellular enzyme CD38 molecules highly expressed on the surface of multiple myeloma and various solid tumor cells, inducing rapid death of tumor cells through a variety of immune-mediated mechanisms, including complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP) as well as through apoptosis. In addition, Darzalex has also been shown to be able to target immunosuppressive cells in the tumor microenvironment and thus exhibit immunomodulatory activity. In addition to multiple myeloma, Darzalex also has the potential to treat other types of tumors that highly express the CD38 molecule, including diffuse large B-cell lymphoma (DLBCL), chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), plasma cell leukemia (PCL), acute myeloid leukemia (AML), follicular lymphoma (FL) and mantle cell lymphoma (MCL).

[0006] Furthermore, the development and use of new anti-CD38 antibodies is needed in the art. Summary of the invention

[0007] The technical problem to be solved by the present invention is to obtain a high-affinity antibody that specifically binds to CD38, especially human CD38, through hybridoma screening and humanization technology.

[0008] In view of the above technical problems, the purpose of the present invention is to provide an antibody or a fragment thereof that specifically binds to CD38, especially human CD38, and based on the antibody or the fragment thereof, provide its use. The "fragment" of the antibody molecule described in the present invention covers various functional fragments of antibodies, such as its antigen-binding portion, such as Fab, F(ab')2 or scFv fragment.

[0009] Specifically, the present invention provides the following technical solutions.

[0010] In one aspect, the present invention provides an antibody molecule or a fragment thereof that binds to human CD38, wherein the antibody molecule or the fragment thereof comprises a heavy chain variable region (VH), wherein the heavy chain variable region comprises a CDR1 (VH-CDR1) selected from SEQ ID NO: 1, SEQ ID NO: 7, SEQ ID NO: 11, SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 26, SEQ ID NO: 31, SEQ ID NO: 41 and SEQ ID NO: 147, a CDR2 (VH-CDR2) selected from SEQ ID NO: 2, SEQ ID NO: 8, SEQ ID NO: 12, SEQ ID NO: 17, SEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO: 32, SEQ ID NO: 37 and SEQ ID NO: 42, and a CDR3 (VH-CDR2) selected from SEQ ID NO: 3, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 18, SEQ ID NO: 22, SEQ ID NO: 28, SEQ ID NO: 33, SEQ ID NO: 38, SEQ ID NO: 49, SEQ ID NO: 50 CDR3 (VH-CDR3) shown in SEQ ID NO:43, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48 and SEQ ID NO:137; and

[0011] The antibody molecule or fragment thereof comprises a light chain variable region (VL), wherein the light chain variable region comprises a CDR1 (VL-CDR1) selected from the group consisting of SEQ ID NO:4, SEQ ID NO:14, SEQ ID NO:23, SEQ ID NO:29, SEQ ID NO:34, SEQ ID NO:39, SEQ ID NO:44, SEQ ID NO:49, SEQ ID NO:136 and SEQ ID NO:148, a CDR2 (VL-CDR2) selected from the group consisting of SEQ ID NO:5, SEQ ID NO:24 and SEQ ID NO:35, and a CDR3 (VL-CDR3) selected from the group consisting of SEQ ID NO:6, SEQ ID NO:10, SEQ ID NO:15, SEQ ID NO:19, SEQ ID NO:25, SEQ ID NO:30, SEQ ID NO:36, SEQ ID NO:40, SEQ ID NO:45, SEQ ID NO:138 and SEQ ID NO:139.

[0012] According to a specific embodiment of the present invention, the heavy chain variable region may comprise a CDR combination (VH-CDR1, VH-CDR2, VH-CDR3) selected from the following:

[0013] (1) VH-CDR1 as shown in SEQ ID NO: 1, VH-CDR2 as shown in SEQ ID NO: 2, and VH-CDR3 as shown in SEQ ID NO: 3;

[0014] (2) VH-CDR1 as shown in SEQ ID NO:7, VH-CDR2 as shown in SEQ ID NO:8, and VH-CDR3 as shown in SEQ ID NO:9;

[0015] (3) VH-CDR1 as shown in SEQ ID NO: 11, VH-CDR2 as shown in SEQ ID NO: 12, and VH-CDR3 as shown in SEQ ID NO: 13;

[0016] (4) VH-CDR1 as shown in SEQ ID NO: 16, VH-CDR2 as shown in SEQ ID NO: 17, and VH-CDR3 as shown in SEQ ID NO: 18;

[0017] (5) VH-CDR1 as shown in SEQ ID NO:20, VH-CDR2 as shown in SEQ ID NO:21, and VH-CDR3 as shown in SEQ ID NO:22;

[0018] (6) VH-CDR1 as shown in SEQ ID NO:26, VH-CDR2 as shown in SEQ ID NO:27, and VH-CDR3 as shown in SEQ ID NO:28;

[0019] (7) VH-CDR1 as shown in SEQ ID NO:31, VH-CDR2 as shown in SEQ ID NO:32, and VH-CDR3 as shown in SEQ ID NO:33;

[0020] (8) VH-CDR1 as shown in SEQ ID NO:26, VH-CDR2 as shown in SEQ ID NO:37, and VH-CDR3 as shown in SEQ ID NO:38;

[0021] (9) VH-CDR1 as shown in SEQ ID NO:41, VH-CDR2 as shown in SEQ ID NO:42, and VH-CDR3 as shown in SEQ ID NO:43;

[0022] (10) VH-CDR1 as shown in SEQ ID NO: 11, VH-CDR2 as shown in SEQ ID NO: 12, and VH-CDR3 as shown in SEQ ID NO: 46;

[0023] (11) VH-CDR1 as shown in SEQ ID NO:11, VH-CDR2 as shown in SEQ ID NO:12, and VH-CDR3 as shown in SEQ ID NO:47;

[0024] (12) VH-CDR1 as shown in SEQ ID NO: 11, VH-CDR2 as shown in SEQ ID NO: 12, and VH-CDR3 as shown in SEQ ID NO: 48;

[0025] (13) VH-CDR1 as shown in SEQ ID NO: 16, VH-CDR2 as shown in SEQ ID NO: 17, and VH-CDR3 as shown in SEQ ID NO: 137;

[0026] (14) VH-CDR1 as shown in SEQ ID NO: 147, VH-CDR2 as shown in SEQ ID NO: 42, and VH-CDR3 as shown in SEQ ID NO: 43;

[0027] And / or, the antibody molecule or fragment thereof comprises a light chain variable region (VL), wherein the light chain variable region may comprise a CDR combination (VL-CDR1, VL-CDR2, VL-CDR3) selected from the following:

[0028] (1) VL-CDR1 as shown in SEQ ID NO:4, VL-CDR2 as shown in SEQ ID NO:5, and VL-CDR3 as shown in SEQ ID NO:6;

[0029] (2) VL-CDR1 as shown in SEQ ID NO:4, VL-CDR2 as shown in SEQ ID NO:5, and VL-CDR3 as shown in SEQ ID NO:10;

[0030] (3) VL-CDR1 as shown in SEQ ID NO: 14, VL-CDR2 as shown in SEQ ID NO: 5, and VL-CDR3 as shown in SEQ ID NO: 15;

[0031] (4) VL-CDR1 as shown in SEQ ID NO:4, VL-CDR2 as shown in SEQ ID NO:5, and VL-CDR3 as shown in SEQ ID NO:19;

[0032] (5) VL-CDR1 as shown in SEQ ID NO:23, VL-CDR2 as shown in SEQ ID NO:24, and VL-CDR3 as shown in SEQ ID NO:25;

[0033] (6) VL-CDR1 as shown in SEQ ID NO:29, VL-CDR2 as shown in SEQ ID NO:24, and VL-CDR3 as shown in SEQ ID NO:30;

[0034] (7) VL-CDR1 as shown in SEQ ID NO:34, VL-CDR2 as shown in SEQ ID NO:35, and VL-CDR3 as shown in SEQ ID NO:36;

[0035] (8) VL-CDR1 as shown in SEQ ID NO:39, VL-CDR2 as shown in SEQ ID NO:24, and VL-CDR3 as shown in SEQ ID NO:40;

[0036] (9) VL-CDR1 as shown in SEQ ID NO:44, VL-CDR2 as shown in SEQ ID NO:24, and VL-CDR3 as shown in SEQ ID NO:45;

[0037] (10) VL-CDR1 as shown in SEQ ID NO:49, VL-CDR2 as shown in SEQ ID NO:24, and VL-CDR3 as shown in SEQ ID NO:40;

[0038] (11) VL-CDR1 as shown in SEQ ID NO:4, VL-CDR2 as shown in SEQ ID NO:5, and VL-CDR3 as shown in SEQ ID NO:138;

[0039] (12) VL-CDR1 as shown in SEQ ID NO: 136, VL-CDR2 as shown in SEQ ID NO: 35, and VL-CDR3 as shown in SEQ ID NO: 36;

[0040] (13) VL-CDR1 as shown in SEQ ID NO: 136, VL-CDR2 as shown in SEQ ID NO: 35, and VL-CDR3 as shown in SEQ ID NO: 139;

[0041] (14) VL-CDR1 as shown in SEQ ID NO:148, VL-CDR2 as shown in SEQ ID NO:24, and VL-CDR3 as shown in SEQ ID NO:45.

[0042] Preferably, the heavy chain variable region in the antibody molecule or fragment thereof comprises an amino acid sequence selected from any one of SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68 to SEQ ID NO:88, SEQ ID NO:140 to SEQ ID NO:142, or an amino acid sequence having at least 75% identity thereto; and / or

[0043] The light chain variable region comprises an amino acid sequence selected from any one of SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:89 to SEQ ID NO:105, SEQ ID NO:134 and SEQ ID NO:135, SEQ ID NO:143 to SEQ ID NO:146, or an amino acid sequence that is at least 75% identical to the amino acid sequence shown.

[0044] According to a specific embodiment of the present invention, the antibody molecule or fragment thereof comprises a CDR combination selected from the following:

[0045] (1) VH-CDR1 as shown in SEQ ID NO: 1, VH-CDR2 as shown in SEQ ID NO: 2, VH-CDR3 as shown in SEQ ID NO: 3; VL-CDR1 as shown in SEQ ID NO: 4, VL-CDR2 as shown in SEQ ID NO: 5, VL-CDR3 as shown in SEQ ID NO: 6;

[0046] (2) VH-CDR1 as shown in SEQ ID NO:7, VH-CDR2 as shown in SEQ ID NO:8, VH-CDR3 as shown in SEQ ID NO:9; VL-CDR1 as shown in SEQ ID NO:4, VL-CDR2 as shown in SEQ ID NO:5, VL-CDR3 as shown in SEQ ID NO:10;

[0047] (3) VH-CDR1 as shown in SEQ ID NO: 11, VH-CDR2 as shown in SEQ ID NO: 12, VH-CDR3 as shown in SEQ ID NO: 13; VL-CDR1 as shown in SEQ ID NO: 14, VL-CDR2 as shown in SEQ ID NO: 5, VL-CDR3 as shown in SEQ ID NO: 15;

[0048] (4) VH-CDR1 as shown in SEQ ID NO: 16, VH-CDR2 as shown in SEQ ID NO: 17, VH-CDR3 as shown in SEQ ID NO: 18; VL-CDR1 as shown in SEQ ID NO: 4, VL-CDR2 as shown in SEQ ID NO: 5, VL-CDR3 as shown in SEQ ID NO: 19;

[0049] (5) VH-CDR1 as shown in SEQ ID NO:20, VH-CDR2 as shown in SEQ ID NO:21, VH-CDR3 as shown in SEQ ID NO:22; VL-CDR1 as shown in SEQ ID NO:23, VL-CDR2 as shown in SEQ ID NO:24, VL-CDR3 as shown in SEQ ID NO:25;

[0050] (6) VH-CDR1 as set forth in SEQ ID NO:26, VH-CDR2 as set forth in SEQ ID NO:27, VH-CDR3 as set forth in SEQ ID NO:28; VL-CDR1 as set forth in SEQ ID NO:29, VL-CDR2 as set forth in SEQ ID NO:24, VL-CDR3 as set forth in SEQ ID NO:30;

[0051] (7) VH-CDR1 as shown in SEQ ID NO:31, VH-CDR2 as shown in SEQ ID NO:32, VH-CDR3 as shown in SEQ ID NO:33; VL-CDR1 as shown in SEQ ID NO:34, VL-CDR2 as shown in SEQ ID NO:35, VL-CDR3 as shown in SEQ ID NO:36;

[0052] (8) VH-CDR1 as set forth in SEQ ID NO:26, VH-CDR2 as set forth in SEQ ID NO:37, VH-CDR3 as set forth in SEQ ID NO:38; VL-CDR1 as set forth in SEQ ID NO:39, VL-CDR2 as set forth in SEQ ID NO:24, VL-CDR3 as set forth in SEQ ID NO:40;

[0053] (9) VH-CDR1 as set forth in SEQ ID NO:41, VH-CDR2 as set forth in SEQ ID NO:42, VH-CDR3 as set forth in SEQ ID NO:43; VL-CDR1 as set forth in SEQ ID NO:44, VL-CDR2 as set forth in SEQ ID NO:24, VL-CDR3 as set forth in SEQ ID NO:45;

[0054] (10) VH-CDR1 as set forth in SEQ ID NO: 11, VH-CDR2 as set forth in SEQ ID NO: 12, VH-CDR3 as set forth in SEQ ID NO: 47; VL-CDR1 as set forth in SEQ ID NO: 14, VL-CDR2 as set forth in SEQ ID NO: 5, VL-CDR3 as set forth in SEQ ID NO: 15;

[0055] (11) VH-CDR1 as set forth in SEQ ID NO: 16, VH-CDR2 as set forth in SEQ ID NO: 17, VH-CDR3 as set forth in SEQ ID NO: 137; VL-CDR1 as set forth in SEQ ID NO: 4, VL-CDR2 as set forth in SEQ ID NO: 5, VL-CDR3 as set forth in SEQ ID NO: 19;

[0056] (12) VH-CDR1 as set forth in SEQ ID NO: 16, VH-CDR2 as set forth in SEQ ID NO: 17, VH-CDR3 as set forth in SEQ ID NO: 137; VL-CDR1 as set forth in SEQ ID NO: 4, VL-CDR2 as set forth in SEQ ID NO: 5, VL-CDR3 as set forth in SEQ ID NO: 138;

[0057] (13) VH-CDR1 as set forth in SEQ ID NO:31, VH-CDR2 as set forth in SEQ ID NO:32, VH-CDR3 as set forth in SEQ ID NO:33; VL-CDR1 as set forth in SEQ ID NO:136, VL-CDR2 as set forth in SEQ ID NO:35, VL-CDR3 as set forth in SEQ ID NO:36;

[0058] (14) VH-CDR1 as set forth in SEQ ID NO: 147, VH-CDR2 as set forth in SEQ ID NO: 42, VH-CDR3 as set forth in SEQ ID NO: 43; VL-CDR1 as set forth in SEQ ID NO: 44, VL-CDR2 as set forth in SEQ ID NO: 24, VL-CDR3 as set forth in SEQ ID NO: 45;

[0059] Preferably, the heavy chain variable region and light chain variable region comprised by the antibody molecule or fragment thereof can be selected from the following combinations:

[0060] (1) the amino acid sequence as set forth in SEQ ID NO:50 or an amino acid sequence having at least 75% identity to the amino acid sequence as set forth in SEQ ID NO:50; and, the amino acid sequence as set forth in SEQ ID NO:51 or an amino acid sequence having at least 75% identity to the amino acid sequence as set forth in SEQ ID NO:51;

[0061] (2) the amino acid sequence as set forth in SEQ ID NO:52 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:52; and, the amino acid sequence as set forth in SEQ ID NO:53 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:53;

[0062] (3) the amino acid sequence as set forth in SEQ ID NO:54 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:54; and, the amino acid sequence as set forth in SEQ ID NO:55 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:55;

[0063] (4) the amino acid sequence as set forth in SEQ ID NO:56 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:56; and, the amino acid sequence as set forth in SEQ ID NO:57 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:57;

[0064] (5) the amino acid sequence as set forth in SEQ ID NO:58 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:58; and, the amino acid sequence as set forth in SEQ ID NO:59 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:59;

[0065] (6) the amino acid sequence as set forth in SEQ ID NO:60 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:60; and, the amino acid sequence as set forth in SEQ ID NO:61 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:61;

[0066] (7) the amino acid sequence as set forth in SEQ ID NO:62 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:62; and, the amino acid sequence as set forth in SEQ ID NO:63 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:63;

[0067] (8) the amino acid sequence as set forth in SEQ ID NO:64 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:64; and, the amino acid sequence as set forth in SEQ ID NO:65 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:65;

[0068] (9) the amino acid sequence as set forth in SEQ ID NO:66 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:66; and, the amino acid sequence as set forth in SEQ ID NO:67 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:67;

[0069] (10) the amino acid sequence as set forth in SEQ ID NO:69 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:69; and, the amino acid sequence as set forth in SEQ ID NO:90 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:90;

[0070] (11) the amino acid sequence as set forth in SEQ ID NO:71 or an amino acid sequence having at least 75% identity to the amino acid sequence as set forth in SEQ ID NO:71; and, the amino acid sequence as set forth in SEQ ID NO:90 or an amino acid sequence having at least 75% identity to the amino acid sequence as set forth in SEQ ID NO:90;

[0071] (12) the amino acid sequence as set forth in SEQ ID NO:74 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:74; and, the amino acid sequence as set forth in SEQ ID NO:92 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:92;

[0072] (13) the amino acid sequence as set forth in SEQ ID NO:75 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:75; and, the amino acid sequence as set forth in SEQ ID NO:94 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:94;

[0073] (14) the amino acid sequence as set forth in SEQ ID NO:76 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:76; and, the amino acid sequence as set forth in SEQ ID NO:94 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:94;

[0074] (15) the amino acid sequence as set forth in SEQ ID NO:77 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:77; and, the amino acid sequence as set forth in SEQ ID NO:95 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:95;

[0075] (16) the amino acid sequence as set forth in SEQ ID NO:83 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:83; and, the amino acid sequence as set forth in SEQ ID NO:102 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:102;

[0076] (17) the amino acid sequence as set forth in SEQ ID NO:84 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:84; and, the amino acid sequence as set forth in SEQ ID NO:102 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:102;

[0077] (18) the amino acid sequence as set forth in SEQ ID NO:85 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:85; and, the amino acid sequence as set forth in SEQ ID NO:134 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:134;

[0078] (19) the amino acid sequence as set forth in SEQ ID NO:88 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:88; and, the amino acid sequence as set forth in SEQ ID NO:103 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:103;

[0079] (20) the amino acid sequence as set forth in SEQ ID NO:88 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:88; and, the amino acid sequence as set forth in SEQ ID NO:104 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO:104;

[0080] (21) the amino acid sequence as set forth in SEQ ID NO: 140 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO: 140; and, the amino acid sequence as set forth in SEQ ID NO: 143 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO: 143;

[0081] (22) the amino acid sequence as set forth in SEQ ID NO: 141 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO: 141; and, the amino acid sequence as set forth in SEQ ID NO: 144 or an amino acid sequence that is at least 75% identical to the amino acid sequence as set forth in SEQ ID NO: 144;

[0082] (23) the amino acid sequence as shown in SEQ ID NO:142 or an amino acid sequence that is at least 75% identical to the amino acid sequence as shown in SEQ ID NO:142; and, the amino acid sequence as shown in SEQ ID NO:144 or an amino acid sequence that is at least 75% identical to the amino acid sequence as shown in SEQ ID NO:144.

[0083] The antibody molecule or fragment thereof of the present invention may be in any form such as a monoclonal antibody, a single-chain antibody, a bifunctional antibody, a single-domain antibody, a nanobody, a fully or partially humanized antibody or a chimeric antibody, or the antibody molecule or fragment thereof may be a half antibody or an antigen-binding fragment of a half antibody, such as scFv, BsFv, dsFv, (dsFv)2, Fab, Fab', F(ab')2 or Fv; with respect to antibody fragments, the antigen-binding fragment of an antibody is particularly preferred.

[0084] Preferably, the antibody molecule or fragment thereof further comprises a human or mouse constant region, preferably a human or mouse light chain constant region (CL) and / or a heavy chain constant region (CH);

[0085] More preferably, the antibody molecule or fragment thereof comprises a heavy chain constant region selected from IgG, IgA, IgM, IgD or IgE and / or a kappa or lambda type light chain constant region.

[0086] According to a specific embodiment of the present invention, the antibody molecule is a monoclonal antibody, preferably a murine, chimeric or humanized monoclonal antibody; preferably, the heavy chain constant region of the monoclonal antibody is of IgG1 or IgG4 subtype, and the light chain constant region is of κ type;

[0087] Preferably, the heavy chain constant region of the monoclonal antibody comprises the amino acid sequence shown in SEQ ID NO: 106 or an amino acid sequence having at least 75% identity with the amino acid sequence;

[0088] Preferably, the light chain constant region of the monoclonal antibody comprises the amino acid sequence shown in SEQ ID NO: 107 or an amino acid sequence that is at least 75% identical to the amino acid sequence.

[0089] As used herein, "at least 75% identity" is, for example, at least 80%, preferably at least 85%, more preferably at least 90%, further preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or even 99% identity, etc., any percentage identity ≥ 75%.

[0090] Based on the antibody molecule or fragment thereof of the present invention, the present invention also provides a conjugate or fusion protein comprising the antibody or fragment thereof of the present invention. The conjugate or fusion protein may include other parts that are bound to the antibody or fragment thereof of the present invention by chemical or physical methods, such as cell surface receptors, small molecule compounds such as amino acids and sugars, small molecule polymers, or any other parts that modify the antibody of the present invention, or even active proteins or polypeptides.

[0091] In another aspect, the present invention also provides a nucleic acid molecule encoding the heavy chain CDR, light chain CDR, light chain variable region, heavy chain variable region, heavy chain or light chain in any antibody or fragment thereof of the present invention.

[0092] In another aspect, the present invention provides a vector comprising the nucleic acid molecule of the present invention. The vector may be a eukaryotic expression vector, a prokaryotic expression vector, an artificial chromosome, a phage vector, etc. The vector or nucleic acid molecule of the present invention may be used to transform or transfect a host cell or enter a host cell in any manner for purposes such as storage or expression of antibodies.

[0093] Therefore, on the other hand, the present invention provides a host cell, the host cell comprises nucleic acid molecules and / or vectors of the present invention, or the host cell is transformed or transfected by nucleic acid molecules and / or vectors of the present invention. The host cell can be any prokaryotic or eukaryotic cell, for example a bacterium or an insect, fungus, plant or animal cell.

[0094] Based on the disclosure of the present invention, the antibody molecules or fragments thereof and the corresponding conjugates or fusion proteins, nucleic acid molecules, vectors and / or host cells provided by the present invention can be obtained by using any conventional technical method known in the art. The antibody molecules or fragments thereof, conjugates or fusion proteins, nucleic acid molecules, vectors and / or host cells can be included in pharmaceutical compositions, more particularly in pharmaceutical preparations, so as to be used for various purposes according to actual needs.

[0095] Therefore, in another aspect, the present invention also provides a pharmaceutical composition, comprising the antibody molecule or fragment thereof, conjugate or fusion protein, nucleic acid molecule, vector and / or host cell described in the present invention, and optional pharmaceutically acceptable excipients.

[0096] In another aspect, the present invention provides use of the above-mentioned antibody molecule or fragment thereof, conjugate or fusion protein, nucleic acid molecule, vector, host cell and / or pharmaceutical composition in the preparation of a drug for treating a disease associated with CD38 expression or mediated by CD38;

[0097] Preferably, the disease is a hematological malignancy, preferably multiple myeloma or non-Hodgkin's lymphoma.

[0098] In yet another aspect, the present invention provides a kit comprising the antibody molecule or fragment thereof, conjugate or fusion protein, nucleic acid molecule, vector, host cell and / or pharmaceutical composition of the present invention.

[0099] In the present invention, hybridoma technology is used to obtain mouse antibodies, and candidate mouse antibodies are obtained through antibody activity analysis (ELISA binding, epitope competition with control antibodies, affinity kinetics); then, the sequences encoding the light and heavy chain variable regions of the candidate mouse antibodies are cloned upstream of the sequences encoding the light and heavy chain constant regions of human antibodies, and mammalian cells are expressed to prepare chimeric antibodies, and then antibody activity analysis is performed to verify the chimeric antibodies' complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) on effector cells, as well as their apoptotic activity on Duadi-luc. Verification; then, according to the Germline database, the humanized template was selected to carry out the humanized design of the antibody sequence. The obtained humanized antibody was again analyzed for antibody activity, and the complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) of the humanized modified antibody on effector cells were repeatedly verified by in vitro cytological experiments, as well as the apoptotic activity of Duadi-luc; then the in vitro physicochemical properties of the humanized antibody were analyzed, and finally a new anti-CD38 humanized antibody sequence with affinity and specificity comparable to or superior to that of the control antibody was obtained.

[0100] Experiments have shown that the antibody molecules provided by the present invention have similar or consistent antigen binding recognition epitopes with known antibodies, and also have the same or better functional activities as known antibodies, including in vitro ADCC, CDC and Daudi cell apoptosis activities. BRIEF DESCRIPTION OF THE DRAWINGS

[0101] The embodiments of the present invention are described in detail below with reference to the accompanying drawings, wherein:

[0102] Figure 1 The results of flow cytometry identification of CD38 expressing cell lines are shown, among which, Figure 1 A: cell line CHOblank; Figure 1 B: cell line 1-T-12; Figure 1 C: Cell line 1-T-14.

[0103] Figure 2 The first round of hybridoma cells is shown ( Figure 2 A) and the second round ( Figure 2 B) Fusion screening results.

[0104] Figure 3 The binding activity of the antibody of the present invention to the antigen CD38 expressed on the cell surface was shown.

[0105] Figure 4 The antibody-dependent cellular cytotoxicity of the antibody of the present invention is shown, wherein Figure 4 A: 8B7 humanized antibody; Figure 4 B: 20M19 humanized antibody; Figure 4 C: 32A7 humanized antibody; Figure 4 D: 5E19 humanized antibody.

[0106] Figure 5 The results of antigen epitope analysis of the antibody of the present invention are shown, wherein: Figure 5 A: Relative to Daratumumab; Figure 5 B: relative to Mor202; Figure 5 C: relative to isatuximab.

[0107] Figure 6 The serum stability results of the antibodies of the present invention are shown, wherein, Figure 6 A: Daratumumab; Figure 6 B: 8B7-H3L3; Figure 6 C:17H13-H2L2; Figure 6 D:20M19-H1L1. DETAILED DESCRIPTION

[0108] The present invention is described below with reference to specific examples. It will be appreciated by those skilled in the art that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

[0109] The experimental methods in the following examples are conventional methods unless otherwise specified. The medicinal materials, reagents, etc. used in the following examples are commercially available products unless otherwise specified.

[0110] The heavy chain constant region sequence of the human monoclonal antibody IgG1 subclass is shown in SEQ ID NO: 106 and SEQ ID NO: 126, and the light chain constant region sequence of the human monoclonal antibody κ subclass is shown in SEQ ID NO: 107 and SEQ ID NO: 127.

[0111] Recombinant human CD38 protein was purchased from ACRO biosystem, Cat No: CD8-H5224.

[0112] Example 1 Synthesis and expression of control antibodies

[0113] The fully synthesized Daratumumab, Isatuximab and Mor202 antibody light chain variable region and heavy chain variable region genes were cloned into eukaryotic expression vectors containing human-kappa light chain constant region and human IgG1 heavy chain constant region encoding genes upstream, and Daratumumab, Isatuximab and Mor202 light and heavy chain expression plasmids were obtained respectively. The obtained plasmids were transferred into Escherichia coli for amplification, and a large amount of plasmids containing Daratumumab, Isatuximab or Mor202 antibody light chain and heavy chain were isolated and obtained, and the two were mixed with PEI and co-transfected into HEK293 cells.

[0114] 5-6 days after cell transfection, the culture supernatant was collected and purified using a Mabselect affinity chromatography column to obtain Daratumumab, Isatuximab and Mor202 antibody recombinant proteins.

[0115] Daratumumab: See SEQ ID NO: 128 to SEQ ID NO: 129 for the sequence.

[0116] Isatuximab: See SEQ ID NO:130 to SEQ ID NO:131 for sequence.

[0117] Mor202: See SEQ ID NO:132 to SEQ ID NO:133 for the sequence.

[0118] Example 2 Preparation of CD38 antigen expressing cell line

[0119] The human CD38 high-expressing CHO cell line 1-T-14 and the human CD38 low-expressing CHO cell line 1-T-12 were prepared.

[0120] The reading frame of the CD38 gene was cloned from a vector containing CD38 cDNA (Beijing Yiqiao Shenzhou, Cat: HG10818-UT) by PCR, and the reading frame of the CD38 gene (the correct sequence has been sequenced and verified) was cloned into a stable expression vector containing a glutamine synthetase (GS) screening gene by enzyme digestion. The CHO-K1 cells in suspension culture were electrotransfected (Nucleofector IIb, Lonza), and the transfected cells were placed in CD CHO AGTTM medium (Gibco, Cat: 12490-025) containing 50 μM MSX (Sigma, Cat: M5379), inoculated into 96-well cell culture plates, and cultured at 37°C, 5% CO2 for 2-3 weeks. After MSX pressure screening, pre-screening under the microscope and amplification culture were performed, and finally two clones (1-T-12 and 1-T-14) were selected by flow cytometry analysis (FACS) for amplification culture and cryopreservation.

[0121] The flow cytometry results of the two cell lines and the negative control CHO cell line (CHO-blank) are shown in Figure 1 The mean fluorescence intensity (MFI) values ​​are shown in Table 1.

[0122] Table 1 Expression of CD38 antigen expressing cell lines (FACS)

[0123] Cell lines MFI CHO blank 391.3 1-T-12 66006.4 1-T-14 133008

[0124] Example 3 Screening and identification of hybridoma cells

[0125] 1. Mouse Immunization

[0126] Balb / c mice aged 8-10 weeks were immunized with Freund's adjuvant and water-soluble adjuvant. Freund's adjuvant was injected intraperitoneally twice on day 0 and day 14, and water-soluble adjuvant was injected intramuscularly twice on day 0 and day 21. The immunization antigen was human CD38 recombinant protein, which was recombinantly expressed in HEK293 cells by Acro biosytem Biotechnology Co., Ltd. The first immunization dose was 50 μg, and the second immunization dose was 25 μg. Before immunization, mouse serum was taken as a negative control for detection. Blood was collected from the tail vein on day 28 after the first immunization with Freund's adjuvant and on day 35 after the first immunization with water-soluble adjuvant. The serum titer was detected by ELISA using a 96-well enzyme-labeled plate coated with recombinant human CD38 protein. Mice with serum titers reaching the fusion requirements were boosted with 25 μg of antigen diluted to 500 μl with D-PBS and injected intraperitoneally. On days 18-38 after the initial immunization, spleen cells from mice with high serum titers were collected for the next step of cell fusion.

[0127] 2. Cell fusion and hybridoma preparation

[0128] 2.1 Myeloma cell preparation

[0129] The myeloma cells P3X63Ag8.653 required for fusion were cultured in a 500 ml triangular culture flask until the cell density reached 0.8-1.0E+6, and the medium was replaced with complete hybridoma culture medium for later use.

[0130] 2.2 Lymph node and B lymphocyte preparation

[0131] Mice with titers that meet the requirements were selected, their eyeballs were removed for blood collection, and the serum was separated as a positive control serum for antibody detection; the spleen of the mice was aseptically removed, and a B lymphocyte suspension was prepared according to conventional methods.

[0132] 2.3 Electro Cell Fusion (ECF)

[0133] Myeloma cells P3X63Ag8.653 and B lymphocytes were mixed at a ratio of 4:1 and then electroporated for fusion. After electroporation, the cells were placed in a 37°C CO2 incubator, left to stand for 30 minutes, centrifuged at 1000RPM at room temperature for 10 minutes, resuspended in 360ml hybridoma selection medium, and plated in a 384-well plate, ensuring that the plate density was 8000-20000 cells / well. The medium was changed once after 2-3 days, and positive hybridoma screening was performed on the 7th-10th day.

[0134] 2.4 Positive hybridoma screening (FACS)

[0135] The fused hybridoma cells were cultured in 384-well plates, and the supernatant was analyzed by FACS to analyze the antibodies secreted by the hybridoma cells. Several clones were screened, which could bind to the human CD38 high-expressing strain 1-T-14 but not to CHO-blank cells. The screened clones were made into single cells by limiting dilution method, and each hybridoma cell clone obtained after 3 rounds secreted only one antibody.

[0136] The results of the first and second rounds of fusion screening are shown in Figure 2 .

[0137] Example 4 Identification of variable region sequences of mouse monoclonal antibodies, preparation of anti-human CD38 chimeric antibodies, and antibody screening

[0138] After the hybridoma cells secreting anti-human CD38 antibodies were expanded and cultured, total cell RNA was extracted according to the instructions of the RNAfast200 kit (Shanghai Feijie Biotechnology Co., Ltd.); the total RNA of the hybridoma cells was reverse transcribed into cDNA using 5× PrimeScript RT Master Mix (Takara); the antibody light chain variable region IgVL (κ) and heavy chain variable region VH sequences were amplified using degenerate primers (Anke Krebber. 1997) and ExtaqPCR reagents (Takara); the PCR amplification products were purified using a PCR clean-upGel extraction kit (Macherey-Nagel); the amplified PCR products were connected to the T vector and transformed into Escherichia coli competent cells according to the instructions of the pClone007 SimpleVector Kit (Qingke Biotechnology Co., Ltd.), and the monoclonal antibody variable region sequences were obtained by DNA sequencing after strain amplification and plasmid extraction.

[0139] The heavy chain variable region sequence of the mouse anti-human CD38 monoclonal antibody and the publicly published heavy chain constant region sequence of the human monoclonal antibody IgG1 subclass were spliced ​​together and constructed into the mammalian cell expression vector pCDNA3.4; the light chain variable region sequence of the mouse anti-human CD38 monoclonal antibody and the publicly published light chain constant region sequence of the human monoclonal antibody κ subclass were spliced ​​together and constructed into the mammalian cell expression vector pCDNA3.1. The constructed heavy chain vector and light chain vector of the anti-human CD38 chimeric antibody were paired and mixed, and HEK293 cells were transfected with polyethyleneimine (PEI). After about 7 days, the cell supernatant was collected and purified using Mabselect to obtain the anti-human CD38 chimeric antibody protein.

[0140] The resulting chimeric antibody is referred to herein as "murine antibody nomenclature-xiIgG".

[0141] The obtained mouse antibodies and chimeric antibodies were subjected to antigen ELISA binding experiments of hybridoma cell culture supernatants or purified antibodies, Octet binding experiments, in vitro cell binding experiments, in vitro cytology experiments (complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC)) and other monoclonal antibody screening (experimental procedures are described below), and the mouse antibodies and the corresponding chimeric antibodies were verified against each other. By combining various experiments, some mouse antibodies and their chimeric antibodies were screened.

[0142] The sequences of the variable regions of the heavy and light chains (VH and VL) from some murine antibodies are shown below, where the antigen complementarity determining clusters (CDRs) of the heavy and light chains are underlined.

[0143] Mouse antibody 2H17:

[0144] >2H17_VH(SEQ ID NO:50)

[0145] QVQLKQSGPGLVQPSQSLSITCTVS GFSLTSYGVH WVRQSPGKGLEWLG VIWRGGSTDYNAAFMS RLSITKDNSKSQVFFKMNSLQADDTAIYYCAK GMITTGYAMDY WGQGTSVTVSS

[0146] >2H17_VL(SEQ ID NO:51)

[0147] DIQMTQSSSSFSVSLGDRVTITC KASEDIYNRLA WYQQKPGNAPRLLIS GATSLET GVPSRFSGSGSGKDYTLSITSLQTEDVATYYC QQYWSTPT FGGGTKLEIK

[0148] Murine antibody 4J1:

[0149] >4J1_VH(SEQ ID NO:52)

[0150] QVQLKQSGPGLVQPSQSLSITCTVS GFSLTSFGIH WVRQSPGKGLEWLG VIWRGGSTDYNAAFLS RLSITKDNSKSQVFFKINSLRADDTAIYFCAK SMITTGYAMDY WGQGTSVTVSS

[0151] >4J1_VL(SEQ ID NO:53)

[0152] DIQMTQSSSSFSVSLGDRVTITC KASEDIYNRLA WYQQKPGNAPRLLIS GATSLET GVPSRFSGSGSGKDYILSITSLQTEDVATYYC QQYWSTPWT FGGGTKLEIK

[0153] Murine antibody 5E19:

[0154] >5E19_VH(SEQ ID NO:54)

[0155] QVQLKQSGPGLVQPSQSLSITCTVS GFSLTNYGVH WLRQSPGKGLEWLG VIWRGGSTDYSAAFMS RLNITKDNSKSQVFFKMNSLQADDTAIYYCAK TTLTRVWYCDV WGAGTTVTVSS

[0156] >5E19_VL(SEQ ID NO:55)

[0157] DIQMTQSSSSFSVSLGDRVTITC KSSEDIYNRLA WYQQKPGNAPRLLIS GATSLET GVPSRFSGSGSGKDYTLSITSLQTEDVVTYYC QQYWNTPLT FGAGTKLELK

[0158] Mouse antibody 8B7:

[0159] >8B7_VH(SEQ ID NO:56)

[0160] QVQLKQSGPGLVQPSQSLSITCTVS GFSLLSYGVH WVRQSPGKGLEWLG VIWRGGSADYNAAFMS RLSITKDNSKSQVFFKMNSLQADDTAIYYCAK SMITTGFTLDY WGQGTSVTVSS

[0161] >8B7_VL(SEQ ID NO:57)

[0162] DIQMTQSSSSFSVSLGDRVTITC KASEDIYNRLA WYQQKPGHAPRLLIS GATSLET GVPSRFSGSGSGKDYTLSITSLQTEDVATYYC QQFWNTPWT FGGGTKLEIK

[0163] Mouse antibody 13D15:

[0164] >13D15_VH(SEQ ID NO:58)

[0165] QIQLVQSGPELKKPGETVKISCKAS GYTFTNFGMN WVKQAPGKGLKWMG WINTYTGEPTNADDFKG RFAFSLETSASTAYLQINNLKNEDMATYFCAR KG WGQGTLVTVSA

[0166] >13D15_VL(SEQ ID NO:59)

[0167] DIVLTQSPASLAVSLGQRTTISC RASESVDSYGNSFMH WYQQKPGQPPKLLIY RASNLES GIPARFSGSGSRTDFTLTINPVEADDVATYYC QQINEDPFT FGSGTKLEIK

[0168] Mouse antibody 15A10:

[0169] >15A10_VH(SEQ ID NO:60)

[0170] QIQLVQSGPELKKPGETVKISCKAS GYTFTNYGMN WVKQAPGKGLKWMG WINTYTAEPTYADDFKG RFAFSLEASASTAYLQINNLKNEDMATYFCAR KYGGY WGQGTTLTVSS

[0171] >15A10_VL(SEQ ID NO:61)

[0172] DIVLTQSPPSLAVSLGQRATISC RASESVDSYGNSFIH WYQQKPGQSPKLLIY RASNLES GIPARFSGSGSRTDFTLTINPVEADDVATYYC QVNEDPLT FGAGTKLELK

[0173] Mouse antibody 17H13:

[0174] >17H13_VH(SEQ ID NO:62)

[0175] QVQLQQSGAELAKPGASVKMSCKAS GYTFTNYWMN WIKQRPGQGLEWIG YIIPSTDYTEYNQKFKD KATLTADKSSGTAYMQLSSLTSEDSAVYYCSR YPYRGYAMDY WGQGTSVTVSS

[0176] >17H13_VL(SEQ ID NO:63)

[0177] QIVLTQSPALMSASPGEKVTMTC SASSSVSYMY WYQQKPRSSPKPWIY LTSNAS GVPARFSGSGSGTSYSLTISSMEAEDAATYYC QQWSTNPWT FGGGTKLELK

[0178] Mouse antibody 20M19:

[0179] >20M19_VH (SEQ ID NO:64)

[0180] QIQLVQSGPELKKPGETVKISCKAS GYTFTNYGMN WVKQAPGKGLKWMG WINTYTGEPTVADDFKG RFAFSLDTSASTAYLQINNLKNEDTATYFCAR NPGWFAY WGQGTLVTVSA

[0181] >20M19_VL (SEQ ID NO:65)

[0182] DIVLTQSPVFLAVSLGQRATISC RASESVDNYGNSFMH WYQQKPGQPPKLLIY RASNLES GIPARFSGSGSRTDFTLTINPVEADDVATYYC QQSHEAPWT FGGGTRLEIK

[0183] Mouse antibody 32A7:

[0184] >32A7_VH (SEQ ID NO:66)

[0185] QIQLVQSGPELKKPGETVKISCKAS GYNLTNYGMN WVKQAPGKGLKWMG WINTYTGEPTYADDFKG RFAFSLETSASTAYLQINNLKNEDTATYFCAR YPGWFAY WGQGTLVTVSA

[0186] >32A7_VL (SEQ ID NO:67)

[0187] DIVLTQSPASLAVSLGQRATISC RTSESVDSYGNSFMH WYQQKPGQPPKLLIY RASNLES GIPARFSGSGSRTDFTLTINPVEADDVATYYC QQQSNEDPWT FGGGTKLEIK

[0188] The results showed that all mouse antibodies with the above-mentioned light and heavy chains had better binding to human CD38 recombinant protein than other mouse antibodies, with obvious effects in cell binding experiments and strong complement-dependent cytotoxicity and antibody-dependent cytotoxicity. Moreover, after being converted into chimeric antibodies, they retained the effects of mouse antibodies, and their binding activity was similar to or higher than that of the control antibodies Daratumumab or Isatuximab.

[0189] Example 5 Humanization of anti-human CD38 mouse antibody and preparation of humanized antibody

[0190] Based on the antibody coding schemes of Kabat and Chothia, the amino acid sequence regions of the six antigen complementarity determining clusters (CDRs) of the heavy and light chains of mouse antibodies and the framework region (framework region) that supports the conservative three-dimensional conformation of the antibody were determined.

[0191] Then, by analyzing and searching known human antibody sequences, we selected the human antibody heavy chain variable region sequence that is most similar to the mouse antibody, such as IGHV1|IGHJ4*01, and selected its antibody framework region sequence as a template to combine the mouse antibody heavy chain CDR with the human antibody framework region to finally generate the humanized antibody heavy chain variable region sequence. The same process was used to generate the humanized antibody light chain variable region sequence.

[0192] In the above process, it was found that antibodies with mouse antibody CDR directly transplanted to human framework regions often show a sharp drop in binding activity, so it is necessary to change individual amino acids in the framework region from human to mouse. When determining the back mutation site, one is to check which amino acids are different by comparing the designed humanized antibody sequence with the original mouse antibody sequence; the second is to check whether these amino acids play an important role in supporting the antibody structure or in binding to the antigen. For the humanized designed sequence, it is also necessary to check whether there are some potential post-translational modification sites, such as N (asparagine) glycosylation site, N deamidation site, D (aspartic acid) isomerization site, etc.

[0193] The humanized antibody heavy chain variable region gene was constructed into a mammalian cell expression vector containing the heavy chain constant region gene of the human monoclonal antibody IgG1 subclass; the light chain variable region gene was constructed into a mammalian cell expression vector containing the light chain constant region gene of the human monoclonal antibody κ subclass. The constructed anti-human CD38 humanized antibody heavy chain vector and light chain vector were paired and mixed, and HEK293 cells were transfected with polyethyleneimine (PEI). After about 7 days, the cell supernatant was collected and purified using Mabselect to obtain the anti-human CD38 humanized antibody protein.

[0194] The partial sequences of the light and heavy chain variable regions obtained by humanization are shown below, wherein the antigen complementarity determining clusters (CDRs) of the heavy and light chains are underlined.

[0195] >5E19_VH_hz0 (SEQ ID NO:68)

[0196] QVQLVESGGGVVQPGRSLRLSCAAS GFSLTNYGVH WVRQAPGKGLEWVA VIWRGGSTDYSAAFMSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR TTLTRVWYCDV WGQGTLVTVSS

[0197] >5E19_VH_hz1(SEQ ID NO:69)

[0198] QVQLVESGGGVVQPGRSLRLSCAAS GFSLTNYGVH WVRQAPGKGLEWVA VIWRGGSTDYSAAFMS RFTISKDNSKNTVYLQMNSLRAEDTAVYYCAK TTLTRVWYCDV WGQGTLVTVSS

[0199] >5E19_VH_hz2(SEQ ID NO:70)

[0200] QVQLVESGGGVVQPGRSLRLSCAVS GFSLTNYGVH WVRQAPGKGLEWVA VIWRGGSTDYSAAFMS RFTISKDNSKNTVYLQMNSLRAEDTAVYYCAK TTLTRVWYFDV WGQGTLVTVSS

[0201] >5E19_VH_hz3(SEQ ID NO:71)

[0202] QVQLVESGGGVVQPGRSLRLSCAVS GFSLTNYGVH WVRQAPGKGLEWVA VIWRGGSTDYSAAFMS RFTISKDNSKNTVYLQMNSLRAEDTAVYYCAK TTLTRVWYVDV WGQGTLVTVSS

[0203] >5E19_VH_hz4(SEQ ID NO:72)

[0204] QVQLVESGGGVVQPGRSLRLSCAVS GFSLTNYGVH WVRQAPGKGLEWVA VIWRGGSTDYSAAFMS RFTISKDNSKNTVYLQMNSLRAEDTAVYYCAK TTLTRVWYYDV WGQGTLVTVSS

[0205] >5E19_VL_hz0(SEQ ID NO:89)

[0206] DIQMTQSPSSLSASVGDRVTITC KSSEDIYNRLA WYQQKPGKAPKLLIY GATSLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYWNTPLT FGQGTKVEIK

[0207] >5E19_VL_hz1(SEQ ID NO:90)

[0208] DIQMTQSPSSLSASVGDRVTITC KSSEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGTDFTLTISSLQPEDVATYYC QQYWNTPLT FGQGTKVEIK

[0209] >8B7_VH_hz0(SEQ ID NO:73)

[0210] QVQLVESGGGVVQPGRSLRLSCAAS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR SMITTGFTL DYWGQGTLVTVSS

[0211] >8B7_VH_hz1(SEQ ID NO:74)

[0212] QVQLVESGGGVVQPGRSLRLSCAAS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISKDNSKNTVYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0213] >8B7_VH_hz3(SEQ ID NO:75)

[0214] QVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISKDNSKSTVYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0215] >8B7_VH_hz4(SEQ ID NO:76)

[0216] EVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMSRFTISKDNSKSTVYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0217] >8B7_VH_hz5(SEQ ID NO:77)

[0218] EVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISKDNSKNTLYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0219] >8B7_VH_hz6(SEQ ID NO:78)

[0220] EVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISRDNSKSTLYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0221] >8B7_VH_hz7(SEQ ID NO:79)

[0222] EVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISRDNSKNTVYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0223] >8B7_VH_hz8(SEQ ID NO:80)

[0224] EVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISKDNSKSTLYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0225] >8B7_VH_hz9(SEQ ID NO:81)

[0226] EVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMSRFTISKDNSKNTVYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0227] >8B7_VH_hz10(SEQ ID NO:82)

[0228] EVQLVESGGGVVQPGRSLRLSCAVS GFSLLSYGVH WVRQAPGKGLEWVA VIWRGGSADYNAAFMS RFTISRDNSKSTVYLQMNSLRAEDTAVYYCAK SMITTGFTL DYWGQGTLVTVSS

[0229] >8B7_VL_hz0(SEQ ID NO:91)

[0230] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC QQFWNTPWT FGPGTKLEIK

[0231] >8B7_VL_hz1(SEQ ID NO:92)

[0232] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIS GATSLET GVPSRFSGSGSGKDYTFTISSLQPEDVATYYC QQFWNTPWT FGPGTKLEIK

[0233] >8B7_VL_hz2(SEQ ID NO:93)

[0234] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC QQFWNTPWT FGQGTKVEIK

[0235] >8B7_VL_hz3(SEQ ID NO:94)

[0236] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIS GATSLETGVPSRFSGSGSGKDYTFTISSLQPEDVATYYC QQFWNTPWT FGQGTKVEIK

[0237] >8B7_VL_hz4(SEQ ID NO:95)

[0238] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGKDFTFTISSLQPEDIATYYC QQFWNTPWTF GQGTKVEIK

[0239] >8B7_VL_hz5(SEQ ID NO:96)

[0240] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGTDYTFTISSLQPEDIATYYC QQFWNTPWTF GQGTKVEIK

[0241] >8B7_VL_hz6(SEQ ID NO:97)

[0242] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGTDFTFTISSLQPEDVATYYC QQFWNTPWTF GQGTKVEIK

[0243] >8B7_VL_hz7(SEQ ID NO:98)

[0244] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGKDYTFTISSLQPEDIATYYC QQFWNTPWTF GQGTKVEIK

[0245] >8B7_VL_hz8(SEQ ID NO:99)

[0246] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLETGVPSRFSGSGSGKDFTFTISSLQPEDVATYYC QQFWNTPWTF GQGTKVEIK

[0247] >8B7_VL_hz9(SEQ ID NO:100)

[0248] DIQMTQSPSSLSASVGDRVTITC KASEDIYNRLA WYQQKPGKAPKLLIY GATSLET GVPSRFSGSGSGTDYTFTISSLQPEDVATYYC QQFWNTPWTF GQGTKVEIK

[0249] >17H13_VH_hz0(SEQ ID NO:83)

[0250] QVQLVQSGAEVKKPGASVKVSCKAS GYTFTNYWMN WVRQAPGQGLEWMG YIIPSTDYTEYNQKFKD RVTMTRDTSTSTVYMELSSLRSEDTAVYYCAR YPYRGYAMDY WGQGTLVTVSS

[0251] >17H13_VH_hz1(SEQ ID NO:84)

[0252] QVQLVQSGAEVKKPGASVKVSCKAS GYTFTNYWMN WVRQAPGQGLEWMG YIIPSTDYTEYNQKFKD RVTMTKDNSTSTAYMELSSLRSEDTAVYYCSR YPYRGYAMDY WGQGTLVTVSS

[0253] >17H13_VH_hz2(SEQ ID NO:85)

[0254] QVQLVQSGAEVKKPGASVKVSCKAS GYTFTNYWMN WVRQAPGQRLEWMG YIIPSTDYTEYNQKFKD RVTITRDTSASTAYMELSSLRSEDTAVYYCAR YPYRGYAMDY WGQGTTVTVSS

[0255] >17H13_VH_hz3(SEQ ID NO:86)

[0256] QVQLVQSGAEVKKPGASVKVSCKAS GYTFTNYWMN WVRQAPGQRLEWMG YIIPSTDYTEYNQKFKDRVTITADKSAGTAYMELSSLRSEDTAVYYCSR YPYRGYAMDY WGQGTTVTVSS

[0257] >17H13_VL_hz0(SEQ ID NO:101)

[0258] EIVLTQSPATLSLSPGERATLSC SASSSVSYMYWY QQKPGQAPRLLIY LTSNLAS GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQWSTNPWT FGQGTKVEIK

[0259] >17H13_VL_hz1(SEQ ID NO:102)

[0260] EIVLTQSPATLSLSPGERATLSC SASSSVSYMYWY QQKPGQAPRPLIY LTSNLAS GIPARFSGSGSGTDYTLTISSLEPEDAAVYYC QQWSTNPWT FGQGTKVEIK

[0261] >17H13_VL_hz2(SEQ ID NO:134)

[0262] QIVLTQSPATLSLSPGERATLTC SASSSVSYMYWY QQKPGQAPRPWIY LTSNLAS GVPARFSGSGSGTSYTLTISSLEPEDFAVYYC QQWSTNPWT FGQGTKVEIK

[0263] >17H13_VL_hz3(SEQ ID NO:135)

[0264] QIVLTQSPATLSLSPGERATLTC SASSSVSYMYWY QQKPGSSPRPWIY LTSNLAS GVPARFSGSGSGTSYTLTISSLEPEDFAVYYC QQWSTNPWTF GGGTKVEIK

[0265] >20M19_VH_hz0(SEQ ID NO:87)

[0266] QVQLVQSGSELKKPGASVKVSCKAS GYTFTNYGMN WVRQAPGQGLEWMG WINTYTGEPTVADDFKGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCAR NPGWFAY WGQGTLVTVSS

[0267] >20M19_VH_hz1(SEQ ID NO:88)

[0268] QIQLVQSGSELKKPGASVKVSCKAS GYTFTNYGMN WVRQAPGQGLEWMG WINTYTGEPTVADDFKG RFVFSLDTSVSTAYLQISSLKAEDTAVYYCAR NPGWFAY WGQGTLVTVSS

[0269] >20M19_VL_hz0(SEQ ID NO:103)

[0270] DIVLTQSPASLAVSPGQRATITC RASESVDNYGNSFMH WYQQKPGQPPKLLIY RASNLES GVPARFSGSGSGTDFTLTINPVEANDTANYYC QQSHEAPWT FGQGTKVEIK

[0271] >20M19_VL_hz1(SEQ ID NO:104)

[0272] DIVLTQSPASLAVSPGQRATITC RASESVDNYGNSFMH WYQQKPGQPPKLLIY RASNLES GVPARFSGSGSRTDFTLTINPVEANDTANYYC QQSHEAPWT FGQGTKVEIK

[0273] >20M19_VL_hz2(SEQ ID NO:105)

[0274] DIVLTQSPASLAVSPGQRATITC RASESVDNYGNTFMH WYQQKPGQPPKLLIY RASNLES GVPARFSGSGSRTDFTLTINPVEAGDTANYYC QQSHEAPWT FGQGTKVEIK

[0275] >32A7_VH_hz0(SEQ ID NO:140)

[0276] QVQLVQSGSELKKPGASVKVSCKAS GYNLTNYGMN WVRQAPGQGLEWMG WINTYTGEPTYADDFKG<h2 style=";text-align:left;direction:ltr">RFVFSLDTSVSTAYLQISSLKAEDTAVYYCAR<h2 style=";text-align:left;direction:ltr"> YPGWFAY <h2 style=";text-align:left;direction:ltr"> WGQGTLVTVSS<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0277] <h2 style=";text-align:left;direction:ltr"> >32A7_VH_hz1(SEQ ID NO:141)<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0278] <h2 style=";text-align:left;direction:ltr"> QIQLVQSGSELKKPGASVKVSCKAS<h2 style=";text-align:left;direction:ltr"> GYNLTNYGMN <h2 style=";text-align:left;direction:ltr"> WVRQAPGQGLEWMG<h2 style=";text-align:left;direction:ltr"> WINTYTGEPTYADDFKG <h2 style=";text-align:left;direction:ltr"> RFVFSLDTSVSTAYLQISSLKAEDTAVYYCAR<h2 style=";text-align:left;direction:ltr"> YPGWFAY <h2 style=";text-align:left;direction:ltr"> WGQGTLVTVSS<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0279] <h2 style=";text-align:left;direction:ltr"> >32A7_VH_hz2(SEQ ID NO:142)<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0280] <h2 style=";text-align:left;direction:ltr"> QIQLVQSGSELKKPGASVKVSCKAS<h2 style=";text-align:left;direction:ltr"> GYALTNYGMN <h2 style=";text-align:left;direction:ltr"> WVRQAPGQGLEWM<h2 style=";text-align:left;direction:ltr"> GWINTYTGEPTYADDFKG <h2 style=";text-align:left;direction:ltr"> RFVFSLDTSVSTAYLQISSLKAEDTAVYYCAR<h2 style=";text-align:left;direction:ltr"> YPGWFAY <h2 style=";text-align:left;direction:ltr"> WGQGTLVTVSS<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0281] <h2 style=";text-align:left;direction:ltr"> >32A7_VL_hz0(SEQ ID NO:143)<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0282] <h2 style=";text-align:left;direction:ltr"> DIVLTQSPASLAVSPGQRATITC<h2 style=";text-align:left;direction:ltr"> RTSESVDSYGNSFMH <h2 style=";text-align:left;direction:ltr"> WYQQKPGQPPKLLIY<h2 style=";text-align:left;direction:ltr"> RASNLES <h2 style=";text-align:left;direction:ltr"> GVPARFSGSGSGTDFTLTINPVEANDTANYYC<h2 style=";text-align:left;direction:ltr"> QQQSNEDPWT <h2 style=";text-align:left;direction:ltr"> FGQGTKVEIK<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0283] <h2 style=";text-align:left;direction:ltr"> >32A7_VL_hz1(SEQ ID NO:144)<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0284] <h2 style=";text-align:left;direction:ltr"> DIVLTQSPASLAVSPGQRATITC<h2 style=";text-align:left;direction:ltr"> RTSESVDSYGNSFMH <h2 style=";text-align:left;direction:ltr"> WYQQKPGQPPKLLIY<h2 style=";text-align:left;direction:ltr"> RASNLES <h2 style=";text-align:left;direction:ltr"> GVPARFSGSGSRTDFTLTINPVEANDTANYYC<h2 style=";text-align:left;direction:ltr"> QQQSNEDPWT <h2 style=";text-align:left;direction:ltr"> FGQGTKVEIK<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0285] <h2 style=";text-align:left;direction:ltr"> >32A7_VL_hz2(SEQ ID NO:145)<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0286] <h2 style=";text-align:left;direction:ltr"> DIVLTQSPASLAVSPGQRATITC<h2 style=";text-align:left;direction:ltr"> RTSESVDSYGNTFMH <h2 style=";text-align:left;direction:ltr"> WYQQKPGQPPKLLIY<h2 style=";text-align:left;direction:ltr"> RASNLESGVPARFSGSGSRTDFTLTINPVEANDTANYYC QQQSNEDPWT FQGKV

[0287] >32A7_VL_hz3 (SEQ ID NO: 146)

[0288] DIVLTQSPASLAVSPGQRATITC RTSESVDSYGNTFMH WYQQKPGQPPKLLIY RASNLES GVPARFSGSGSRTDFTLTINPVEAQDTANYYC QQQSNEDPWT FQGKV

[0289] The humanized antibody was named "Mouse Antibody Naming-HmLn", where m and n are the numbers of the humanized (hz) modified sequences of VH and VL (VH_hz and VL_hz), respectively.

[0290] Example 6 In vitro validation of anti-human CD38 antibody binding to antigen and in vitro cytological assay

[0291] 1. Octet binding assay

[0292] Octet binding experiments were performed on the antibodies of the present invention and the control antibody Daratumumab (Dara) and recombinant human CD38 protein.

[0293] The antibody affinity was determined by the anti-human antibody capture method using the Fortebio (BLITZ pro1.1.0.28) instrument. During the determination, the capture antibody (AHC) biological probe of the anti-human antibody Fc segment was soaked in PBS for 10 minutes; 200 μl of the diluted antibody sample (including the chimeric antibody, humanized antibody and control antibody Daratumumab of the present invention; the antibody working concentration was 15 μg / mL) was loaded onto the AHC biological probe, and then balanced in PBS for 100 seconds, and the AHC probe was further combined with human CD38 protein (ACRO biosystem, Cat No.CD8-H5224) of different dilution concentrations for a binding reaction of 600 seconds, and then the AHC probe was transferred to PBS for a dissociation reaction of 600 seconds. After the experiment was completed, the blank control response value was deducted, and the 1:1 Langmuir binding mode fitting was performed using the software to calculate the kinetic constants of antigen-antibody binding.

[0294] The results are shown in Tables 2 to 5.

[0295] Table 2 In vitro binding activity of antibodies to recombinant human CD38 protein

[0296] Antibody Concentration (nM) response KD(M) kon(1 / Ms) kdis(1 / s) RMax 4J1-xiIgG 100 0.4598 3.31E-08 2.85E+05 9.44E-03 0.6097 4J1-H0L0 100 -0.01 - 2.32E+05 - 0.0001 4J1-H0L1 100 -0.0089 - 1.09E+06 - 0.0001 Dara 100 0.2098 2.44E-07 4.53E+04 1.11E-02 0.7031

[0297] Table 3 In vitro binding activity of antibodies to recombinant human CD38 protein

[0298] Antibody Concentration (nM) response KD(M) kon(1 / Ms) kdis(1 / s) RMax 5E19-xiIgG 100 0.3799 2.79E-08 4.70E+05 1.31E-02 0.4941 5E19-H0L0 100 -0.0155 - 8.33E+04 - 0.0032 5E19-H0L1 100 0.0184 - 2.26E+05 5.13E-01 0.5128 5E19-H1L0 100 0.1096 1.30E-06 6.44E+04 8.39E-02 1.6077 5E19-H1L1 100 0.338 7.76E-08 3.17E+05 2.46E-02 0.6181 Dara 100 0.2225 1.60E-07 7.29E+04 1.16E-02 0.5712

[0299] Table 4 In vitro binding activity of antibodies to recombinant human CD38 protein

[0300]

[0301]

[0302] Table 5 In vitro binding activity of antibodies to recombinant human CD38 protein

[0303] Antibody Concentration (nM) response KD(M) kon(1 / Ms) kdis(1 / s) RMax 17H13-xiIgG 100 0.2662 2.29E-08 2.87E+05 6.56E-03 0.3266 17H13-H0L0 100 -0.0063 - 1.96E+06 - 0.0001 17H13-H0L1 100 0.2254 6.16E-08 1.64E+05 1.01E-02 0.3658 17H13-H1L0 100 0.0053 - 8.49E+03 7.90E-01 8.1272 17H13-H1L1 100 0.2275 3.94E-08 3.08E+05 1.21E-02 0.3203 Dara 100 0.2159 1.43E-07 8.83E+04 1.26E-02 0.5239

[0304] Experimental results showed that some humanized antibodies retained the binding activity to human CD38 recombinant protein.

[0305] 2. In vitro cell binding assay

[0306] An in vitro cell binding experiment of the antibody of the present invention and the control antibody Daratumumab (Dara) to CD38 protein was conducted.

[0307] The anti-human CD38 antibody was diluted 2-fold from a starting concentration of 100 nM, for a total of 16 concentration points. 10 μl of the antibody at each concentration point was added to a 384-well plate. The CHO cell line 1-T-14 expressing CD38 on the cell surface was taken, and the cells were washed once with PBS containing 0.5% BSA, centrifuged at 100 g for 5 minutes at room temperature, and the cells were resuspended at a density of about 2x10 6 cells / ml, take 10μl and add it to the wells of the 384-well plate with antibody. After incubation at 4℃ for 1 hour, add fluorescently labeled goat anti-human IgG secondary antibody. After incubation at 4℃ for another 1 hour, analyze the average fluorescence intensity of the cell population using flow cytometry.

[0308] Results Figure 3 and Table 6.

[0309] Table 6 Binding activity of antibodies and antigens

[0310] Antibody EC50(μg / ml) 8B7-H3L3 0.7368 5E19-H3L1 0.9533 20M19-H1L0 1.101 20M19-H1L1 1.221 Dara 1.279 8B7-H1L1 1.375 Unrelated antibodies -

[0311] 3. In vitro cytological assay

[0312] 3.1 Complement-dependent cytotoxicity (CDC)

[0313] The ability of the antibodies of the present invention and the control antibody Daratumumab (Dara) to induce complement-dependent cytotoxicity (CDC) against the CHO cell line 1-T-14 stably expressing human CD38 was analyzed using commercial human serum full complement from Quidel.

[0314] The cells were mixed with antibodies and control antibodies at final concentrations of 50 μg / ml to 7.6 ng / ml, and then 6.25% human serum complete complement dissolved in cell culture medium RPMI-1640 was added to the cells and incubated at 37°C for 3 hours. Thereafter, cytotoxicity was detected using a CCK-8 kit, and finally the absorbance at 450 nm was detected using an MD microplate reader. The absorbance value was used to perform a 4-parameter curve fitting using softmax pro7 software to calculate the EC50 of the sample.

[0315] The results are shown in Table 7.

[0316] Table 7 Complement dependent cytotoxicity of antibodies

[0317] Plate No. Antibody EC50(ng / mL) Relative activity (%) P1 Dara 154.5 100.0 P1 8B7-H1L1 177.4 87.1 P1 8B7-H3L3 214.5 72.0 P2 Dara 144.5 100.0 P2 20M19-H1L0 234.9 61.5 P2 20M19-H1L1 239.6 60.3 P3 Dara 126.4 100.0 P3 32A7-H1L1 303.2 41.7 P3 32A7-H2L1 203.5 62.1 P4 Dara 125.9 100.0 P4 5E19-H3L1 192.6 65.4 P4 Unrelated antibodies ～ ～

[0318] 3.2 Antibody-dependent cellular cytotoxicity (ADCC)

[0319] The antibodies of the present invention and the control antibody Daratumumab (Dara) were tested for inducing antibody-dependent cellular cytotoxicity (ADCC) against the CHO cell line 1-T-14 stably expressing human CD38.

[0320] Engineered Jurkat cells stably expressing the FcγRIIIa-FcεRIaγ hybrid receptor and firefly luciferase driven by the NFAT response element were used as effector cells. The biological activity of the antibody in the ADCC mechanism of action was quantified by the production of luciferase by the activation of the NFAT pathway.

[0321] 1.5E5 effector cells were mixed with the antibody of the present invention and the control antibody Daratumumab at a final concentration of 33 μg / ml to 85 pg / ml, and then 2.5E4 target cells, i.e., CHO cell line 1-T-14 stably expressing human CD38 (the E:T ratio of effector cells to target cells was 6:1), were added thereto and incubated at 37°C for 16 hours. TM The Luciferase Assay System was used for detection, and the LUM value was finally detected by MD microplate reader, and the induction fold and EC50 were calculated using the following formula.

[0322] Induction fold = (test well reading – background value) / (negative control well reading – background value)

[0323] The induction fold was calculated using prism7 software for a 4-parameter fitting curve and the EC50 of the sample was calculated.

[0324] Results Figure 4 and Table 8.

[0325] Table 8 Antibody-dependent cellular cytotoxicity of antibodies

[0326] Plate No. Antibody EC50(ng / mL) Relative activity (%) P1 Dara 31.76 100.0 P1 8B7-H1L1 60.59 52.4 P1 8B7-H3L3 37.94 83.7 P2 Dara 39.01 100.0 P2 20M19-H1L0 31.55 123.6 P2 20M19-H1L1 41.63 93.7 P3 Dara 32.59 100.0 P3 32A7-H1L1 60.23 54.1 P3 32A7-H2L1 56.42 57.8 P4 Dara 31.61 100.0 P4 5E19-H3L1 51.46 61.4 P4 Unrelated antibodies ～ ～

[0327] Example 9 In vitro binding affinity and kinetics experiments of humanized anti-human CD38 antibodies

[0328] The antibody-antigen interaction was determined using the GE BIAcore S200 instrument.

[0329] Referring to the operating instructions of GE's Biotin CAPture Kit (Cat. No. 28-9202-34, Lot. 10265137), firstly, the His-tagged human CD38 protein antigen was coupled to both the CAP analysis channel and the control sample channel of the sensor chip, and then the gradient diluted antibody sample (starting concentration 20nM, 1:3 dilution for 8 concentration points, and setting the 0.741nm concentration point to repeat) was flowed through the analysis channel and the control sample channel together to measure the light reaction value after the antibody-antigen binding. After fitting analysis by the instrument software, the binding constant Kon and dissociation constant Koff of the antibody, as well as the affinity constant KD, were finally obtained.

[0330] The results are shown in Table 9.

[0331] Table 9 Binding kinetics of humanized antibodies

[0332] ka(1 / Ms) kd(1 / s) KD(M) 20M19-H1L1 1.049E+6 7.136E-4 6.804E-10 8B7-H3L3 6.623E+5 0.001197 1.808E-9 8B7-H4L3 1.277E+6 0.002919 2.287E-9

[0333] Example 10 Determination of apoptotic activity of humanized anti-human CD38 antibody

[0334] Materials: Daudi-Luc (ATCC, Cat No. CCL-213) cells

[0335] Kit: Alex FluorTM 488 Annexin V / Dead Cell Apoptosis Kit (invitrogen, Cat: V13245, Lot: 1923636)

[0336] Tool antibody: Affinipure F(ab')2 Fragment Goat-anti-human IgG Fcr Fragment Specific (Immo Research, Cat: 109-006-008, Lot: 129913, Conc. 1.3 mg / ml)

[0337] Experimental steps:

[0338] Wash the Daudi cells once with pre-cooled PBS, then seed the cells into a 96-well plate at 1E5 / well, add 0.7nM anti-CD38 antibody and incubate for 30 minutes. Add 5μg / ml goat anti-human tool antibody to the 96-well plate and incubate at 4℃ for 24 hours. Wash the cells twice with pre-cooled PBS, discard the supernatant, and resuspend the cells with 100μl 1x Annexin solution. Add 5μl Alexa 488annexin V and 1μL 100μg / mL PI working solution were added to a 96-well plate and incubated at room temperature in the dark for 15 minutes. After the reaction was terminated with 400μl 1Xannexin binding solution, the 96-well plate was placed on ice and the fluorescence excitation 530nM and emission 488nM readings were measured using a flow cytometer.

[0339] The results are shown in Table 10.

[0340] Table 10 Apoptotic activity of the antibodies of the present invention

[0341]

[0342]

[0343] The "cross-linked" and "non-cross-linked" in the table are the forms of bridged Fab and non-bridged Fab, respectively.

[0344] Embodiment 11 Characterization of drugability of humanized anti-human CD38 antibody

[0345] 1. Antibody epitope analysis

[0346] The antigen epitope analysis of the antibody of the present invention was performed using the In tandem method of Fortebio. In short, the monoclonal supernatant containing the mouse antibody of the present invention was first used to bind to the antigen cell, i.e., the human CD38 highly expressing CHO cell line 1-T-14, and then the control antibody was added to compete with the mouse antibody for binding to the antigen epitope. The binding signal of the labeled control antibody was detected. The lower the binding signal of the control antibody, the more it can prove that the mouse antibody occupies the antigen epitope recognized by the control antibody, and the antigen binding epitopes of the antibody of the present invention and the control antibody are close to or consistent.

[0347] The competition results of the antibody of the present invention with Daratumumab, Mor202, and Isatuximab are shown in Figure 5 .

[0348] Experiments have shown that the binding site of the antibody of the present invention to the antigen is consistent with the epitope of the control antibody.

[0349] 2. Antibody Monomer Rate Analysis

[0350] Experimental instrument: UPLC CLASS ACQUITYH (WATERS)

[0351] Analytical column: TSKgel G3000SWXL 7.8*300 (TOSHI, Cat No 003C03326C

[0352] Assay solution: DPBS (GIBCO, Cat No. 14190-136, Lot # 1967705)

[0353] Analysis method: Inject 50 μl of 1 mg / ml antibody into a pre-equilibrated chromatography column, flow at room temperature and a flow rate of 0.7 ml / min for 35 min, and simultaneously detect the absorbance values ​​of machines A280 and A214. Determine the monomer content and ratio of the antibody based on the peak time and peak volume.

[0354] The results are shown in Table 11.

[0355] Table 11 Main peak retention time and monomer rate of the antibodies of the present invention

[0356]

[0357] 3. Analysis of Antibody Hydrophobicity

[0358] Experimental instrument: ARC (Waters)

[0359] Analytical column for experiment: TSKgel Butyl-NPR (4.6mmX3.5cm, Cat No 14947)

[0360] Assay solution: a. 20 mM Histidine, pH 6.0; b. 20 mM Histidine, 1.6 M (NH4)2SO4

[0361] Analysis method: Analyze the hydrophobic properties of the antibody according to the instructions for use of the hydrophobic chromatography column.

[0362] The results are shown in Table 12.

[0363] Table 12 Hydrophobic properties of the antibodies of the present invention

[0364]

[0365]

[0366] 4. Antibody in vitro monkey (cyno) serum stability study

[0367] Experimental materials: antibody to be tested, FBS, antigen human CD38 recombinant protein, anti-huIgG Fab monoclonal antibody (Sigma, I5260-1ML), HRP-labeled goat anti-human IgG secondary antibody (Jackson, code: 109-035-098).

[0368] Experimental instruments: 37℃ incubator, ELISA reader.

[0369] Experimental steps:

[0370] Sample preparation:

[0371] 1) Adjust the concentration of the antibody to be tested to 20 μg / ml, filter and sterilize, and dispense into 250 μl / tube for later use;

[0372] 2) Add an equal volume of FBS to the aliquoted antibody to be tested, i.e., the final concentration is 50% of the serum concentration and 10 μg / ml antibody concentration;

[0373] 3) Prepare 7 samples in total, seal with sealing film, and place at 37°C. Keep sterile throughout the process;

[0374] 4) Take samples on day 0, 3, 7, 10, 14 and 21 and store them at 4°C for testing. The sample taken on day 21 should be kept at 4°C for at least one day.

[0375] Detection method:

[0376] 1) PBS was used to coat the antigen and anti-IgG Fab monoclonal antibody in a 96-well ELISA plate, 0.2 μg / ml, 100 μl / well, at 4°C overnight;

[0377] 2) Prepare the required reagents:

[0378] Blocking solution: 5% BSA + PBS

[0379] Antibody diluent 5% BSA + PBS + 50% FBS

[0380] ELISA plate washing solution 0.1% Tween + PBS

[0381] 3) Wash the coated ELISA plate three times with PBS, 300 μl / well, to remove free uncoated antigens;

[0382] 4) Add blocking solution, 200 μl / well, and block at 37°C for 1 h;

[0383] 5) The antibody to be tested was diluted to 2 μg / ml using antibody diluent and diluted 3-fold in a total of 8 gradients;

[0384] 6) Pour off the blocking solution, add the diluted antibodies to the two coating methods, 100 μl / well, and incubate at 37°C for 1 hour;

[0385] 7) Wash the plate 3 times with PBST;

[0386] 8) Dilute the secondary antibody at 1:5000, add to the washed ELISA plate, 100 μl / well, and incubate at 37°C for 40 min;

[0387] 9) Wash the plate 3 times with PBST;

[0388] 10) TMB color development, 100 μl / well, protected from light for 10 min;

[0389] 11) Add 50 μl 2M HCL to stop the reaction and read at 450 nm.

[0390] Result processing:

[0391] The binding curve was developed according to the ELISA color development value, and the changes in the binding curve after different storage times were observed to evaluate the stability of the antibody binding activity after storage.

[0392] Results Figure 6 .

[0393] 5. Analysis of drug metabolism in antibody mice

[0394] Experimental materials: antibody to be tested, serum collected from mice at different time points, binding antigen human CD38 of the antibody to be tested, antibody standard Dara, anti-huIgG Fab monoclonal antibody (Sigma, I5260-1ML), HRP-labeled goat anti-human IgG secondary antibody (Jackson, code: 109-035-098).

[0395] Experimental methods:

[0396] Serum Collection:

[0397] 1) Female Balb / C mice, 3 mice / group, 200 μg of the antibody to be tested / mouse were administered via the tail vein or intraperitoneal cavity;

[0398] 2) Blood samples were collected from the tail vein at the time points designed for the experiment, placed at room temperature for more than 30 minutes, and serum was collected at 4000 rpm for 15 minutes and stored at -20°C. To prevent serum evaporation, the final serum collection volume should be greater than 20 μl as much as possible;

[0399] 3) The last collected serum should be frozen at -20℃ for at least 24h.

[0400] Detection method:

[0401] 1) PBS was used to coat the antigen and anti-IgG Fab monoclonal antibody in a 96-well ELISA plate, 0.2 μg / ml, 100 μl / well, at 4°C overnight;

[0402] 2) Prepare the required reagents:

[0403] Blocking solution: 5% BSA + PBS

[0404] Antibody diluent 5% BSA + PBS + 20% blank mouse serum

[0405] ELISA plate washing solution 0.1% Tween + PBS

[0406] 3) Wash the coated ELISA plate three times with PBS, 300 μl / well;

[0407] 4) Add blocking solution, 200 μl / well, and block at 37°C for 1 h;

[0408] 5) The starting serum is diluted to an appropriate concentration using blocking solution, and then diluted to an appropriate concentration range using antibody diluent containing the same serum concentration. The specific dilution multiple needs to be adjusted based on preliminary experiments. In principle, the final color value of the tested serum should be within the color value range of the standard curve.

[0409] 6) Dilute the antibody standard with antibody diluent. The dilution of the standard is still adjusted according to the preliminary experiment so that the standard can fit a linear curve (if there is suitable software, an S-shaped curve can also be fitted).

[0410] 7) Pour off the blocking solution, add the diluted antibody standard and the test serum to the ELISA plates of the two coating methods, 100 μl / well, and incubate at 37°C for 1 hour;

[0411] 8) Wash the plate 3 times with PBST;

[0412] 9) Dilute the secondary antibody at 1:5000, add to the washed ELISA plate, 100 μl / well, and incubate at 37°C for 40 min;

[0413] 10) Wash the plate 3 times with PBST;

[0414] 11) TMB color development, 100 μl / well, protected from light for 10 min;

[0415] 12) Add 50 μl 2M HCL to stop the reaction and read at 450 nm.

[0416] The results are shown in Table 13.

[0417] Table 13-1 Drug metabolism properties of the antibodies of the present invention in mice

[0418]

[0419] Table 13-2 Drug metabolism properties of the antibodies of the present invention in mice

[0420]

[0421]

[0422] The above description of the specific embodiments of the present invention does not limit the present invention. Those skilled in the art may make various changes or modifications based on the present invention. As long as they do not depart from the spirit of the present invention, they should all fall within the scope of the claims attached to the present invention. Sequence Listing <110> Maiwei (Shanghai) Biotechnology Co., Ltd. Shanghai PuMing Biotechnology Co., Ltd. <120> Anti-human CD38 antibody and its application <130> LC19110036 <160> 148 <170> SIPOSequenceListing 1.0 <210> 1 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 1 Gly Phe Ser Leu Thr Ser Tyr Gly Val His 1 5 10 <210> 2 <211> 16 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 2 Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Asn Ala Ala Phe Met Ser 1 5 10 15 <210> 3 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 3 Gly Met Ile Thr Thr Gly Tyr Ala Met Asp Tyr 1 5 10 <210> 4 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 4 Lys Ala Ser Glu Asp Ile Tyr Asn Arg Leu Ala 1 5 10 <210> 5 <211> 7 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR2 <400> 5 Gly Ala Thr Ser Leu Glu Thr 1 5 <210> 6 <211> 8 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 6 Gln Gln Tyr Trp Ser Thr Pro Thr 1 5 <210> 7 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 7 Gly Phe Ser Leu Thr Ser Phe Gly Ile His 1 5 10 <210> 8 <211> 16 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 8 Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Asn Ala Ala Phe Leu Ser 1 5 10 15 <210> 9 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 9 Ser Met Ile Thr Thr Gly Tyr Ala Met Asp Tyr 1 5 10 <210> 10 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 10 Gln Gln Tyr Trp Ser Thr Pro Trp Thr 1 5 <210> 11 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 11 Gly Phe Ser Leu Thr Asn Tyr Gly Val His 1 5 10 <210> 12 <211> 16 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 12 Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met Ser 1 5 10 15 <210> 13 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 13 Thr Thr Leu Thr Arg Val Trp Tyr Cys Asp Val 1 5 10 <210> 14 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 14 Lys Ser Ser Glu Asp Ile Tyr Asn Arg Leu Ala 1 5 10 <210> 15 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 15 Gln Gln Tyr Trp Asn Thr Pro Leu Thr 1 5 <210> 16 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 16 Gly Phe Ser Leu Leu Ser Tyr Gly Val His 1 5 10 <210> 17 <211> 16 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 17 Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met Ser 1 5 10 15 <210> 18 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 18 Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr 1 5 10 <210> 19 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 19 Gln Gln Phe Trp Asn Thr Pro Trp Thr 1 5 <210> 20 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 20 Gly Tyr Thr Phe Thr Asn Phe Gly Met Asn 1 5 10 <210> twenty one <211> 17 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> twenty one Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Asn Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> twenty two <211> 5 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> twenty two Lys Gly Phe Ala Tyr 1 5 <210> twenty three <211> 15 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> twenty three Arg Ala Ser Glu Ser Val Asp Ser Tyr Gly Asn Ser Phe Met His 1 5 10 15 <210> twenty four <211> 7 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR2 <400> twenty four Arg Ala Ser Asn Leu Glu Ser 1 5 <210> 25 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 25 Gln Gln Ile Asn Glu Asp Pro Phe Thr 1 5 <210> 26 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 26 Gly Tyr Thr Phe Thr Asn Tyr Gly Met Asn 1 5 10 <210> 27 <211> 17 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 27 Trp Ile Asn Thr Tyr Thr Ala Glu Pro Thr Tyr Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 28 <211> 5 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 28 Lys Tyr Gly Gly Tyr 1 5 <210> 29 <211> 15 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 29 Arg Ala Ser Glu Ser Val Asp Ser Tyr Gly Asn Ser Phe Ile His 1 5 10 15 <210> 30 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 30 Gln Gln Val Asn Glu Asp Pro Leu Thr 1 5 <210> 31 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 31 Gly Tyr Thr Phe Thr Asn Tyr Trp Met Asn 1 5 10 <210> 32 <211> 17 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 32 Tyr Ile Ile Pro Ser Thr Asp Tyr Thr Glu Tyr Asn Gln Lys Phe Lys 1 5 10 15 Asp <210> 33 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 33 Tyr Pro Tyr Arg Gly Tyr Ala Met Asp Tyr 1 5 10 <210> 34 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 34 Ser Ala Ser Ser Ser Val Ser Tyr Met Tyr 1 5 10 <210> 35 <211> 7 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR2 <400> 35 Leu Thr Ser Asn Leu Ala Ser 1 5 <210> 36 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 36 Gln Gln Trp Ser Thr Asn Pro Trp Thr 1 5 <210> 37 <211> 17 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 37 Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Val Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 38 <211> 7 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 38 Asn Pro Gly Trp Phe Ala Tyr 1 5 <210> 39 <211> 15 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 39 Arg Ala Ser Glu Ser Val Asp Asn Tyr Gly Asn Ser Phe Met His 1 5 10 15 <210> 40 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 40 Gln Gln Ser His Glu Ala Pro Trp Thr 1 5 <210> 41 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 41 Gly Tyr Asn Leu Thr Asn Tyr Gly Met Asn 1 5 10 <210> 42 <211> 17 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR2 <400> 42 Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 43 <211> 7 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 43 Tyr Pro Gly Trp Phe Ala Tyr 1 5 <210> 44 <211> 15 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 44 Arg Thr Ser Glu Ser Val Asp Ser Tyr Gly Asn Ser Phe Met His 1 5 10 15 <210> 45 <211> 9 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 45 Gln Gln Ser Asn Glu Asp Pro Trp Thr 1 5 <210> 46 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 46 Thr Thr Leu Thr Arg Val Trp Tyr Phe Asp Val 1 5 10 <210> 47 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 47 Thr Thr Leu Thr Arg Val Trp Tyr Val Asp Val 1 5 10 <210> 48 <211> 11 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 48 Thr Thr Leu Thr Arg Val Trp Tyr Tyr Asp Val 1 5 10 <210> 49 <211> 15 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 49 Arg Ala Ser Glu Ser Val Asp Asn Tyr Gly Asn Thr Phe Met His 1 5 10 15 <210> 50 <211> 119 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 50 Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Leu Ser Ile Thr Lys Asp Asn Ser Lys Ser Gln Val Phe Phe 65 70 75 80 Lys Met Asn Ser Leu Gln Ala Asp Asp Thr Ala Ile Tyr Tyr Cys Ala 85 90 95 Lys Gly Met Ile Thr Thr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 51 <211> 106 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 51 Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Phe Ser Val Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Leu Ser Ile Thr Ser Leu Gln Thr 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 52 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 52 Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Ser Phe 20 25 30 Gly Ile His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Asn Ala Ala Phe Leu 50 55 60 Ser Arg Leu Ser Ile Thr Lys Asp Asn Ser Lys Ser Gln Val Phe Phe 65 70 75 80 Lys Ile Asn Ser Leu Arg Ala Asp Asp Thr Ala Ile Tyr Phe Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 53 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 53 Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Phe Ser Val Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Ile Leu Ser Ile Thr Ser Leu Gln Thr 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 54 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 54 Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Leu Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met 50 55 60 Ser Arg Leu Asn Ile Thr Lys Asp Asn Ser Lys Ser Gln Val Phe Phe 65 70 75 80 Lys Met Asn Ser Leu Gln Ala Asp Asp Thr Ala Ile Tyr Tyr Cys Ala 85 90 95 Lys Thr Thr Leu Thr Arg Val Trp Tyr Cys Asp Val Trp Gly Ala Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 55 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 55 Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Phe Ser Val Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Leu Ser Ile Thr Ser Leu Gln Thr 65 70 75 80 Glu Asp Val Val Thr Tyr Tyr Cys Gln Gln Tyr Trp Asn Thr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 56 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 56 Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Leu Ser Ile Thr Lys Asp Asn Ser Lys Ser Gln Val Phe Phe 65 70 75 80 Lys Met Asn Ser Leu Gln Ala Asp Asp Thr Ala Ile Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 57 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain variable region <400> 57 Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Phe Ser Val Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly His Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Leu Ser Ile Thr Ser Leu Gln Thr 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 58 <211> 114 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 58 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Phe 20 25 30 Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Asn Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Met Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Lys Gly Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 100 105 110 Ser Ala <210> 59 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 59 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Thr Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Ser Tyr 20 25 30 Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Glu Asp Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 60 <211> 114 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 60 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Ala Glu Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Ala Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Met Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Lys Tyr Gly Gly Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val 100 105 110 Ser Ser <210> 61 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 61 Asp Ile Val Leu Thr Gln Ser Pro Pro Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Ser Tyr 20 25 30 Gly Asn Ser Phe Ile His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Val Asn 85 90 95 Glu Asp Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 110 <210> 62 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 62 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met Asn Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Ile Pro Ser Thr Asp Tyr Thr Glu Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Gly Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Tyr Pro Tyr Arg Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 63 <211> 106 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain variable region <400> 63 Gln Ile Val Leu Thr Gln Ser Pro Ala Leu Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Arg Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Thr Asn Pro Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 64 <211> 116 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 64 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Val Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Asn Pro Gly Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ala 115 <210> 65 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 65 Asp Ile Val Leu Thr Gln Ser Pro Val Phe Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser His 85 90 95 Glu Ala Pro Trp Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys 100 105 110 <210> 66 <211> 116 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 66 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Asn Leu Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Tyr Pro Gly Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ala 115 <210> 67 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 67 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Thr Ser Glu Ser Val Asp Ser Tyr 20 25 30 Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Asn 85 90 95 Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 68 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 68 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Thr Thr Leu Thr Arg Val Trp Tyr Cys Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 69 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 69 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Thr Thr Leu Thr Arg Val Trp Tyr Cys Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 70 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 70 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Thr Thr Leu Thr Arg Val Trp Tyr Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 71 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 71 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Thr Thr Leu Thr Arg Val Trp Tyr Val Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 72 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 72 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Thr Thr Leu Thr Arg Val Trp Tyr Tyr Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 73 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 73 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 74 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 74 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 75 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 75 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Ser Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 76 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 76 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Ser Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 77 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 77 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 78 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 78 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Ser Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 79 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 79 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 80 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 80 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Ser Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 81 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 81 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 82 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 82 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ser Leu Leu Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Arg Gly Gly Ser Ala Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Ser Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Ser Met Ile Thr Thr Gly Phe Thr Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 83 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 83 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Ile Pro Ser Thr Asp Tyr Thr Glu Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Pro Tyr Arg Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 84 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 84 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Ile Pro Ser Thr Asp Tyr Thr Glu Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Lys Asp Asn Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Tyr Pro Tyr Arg Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 85 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 85 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Tyr Ile Ile Pro Ser Thr Asp Tyr Thr Glu Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Pro Tyr Arg Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 86 <211> 119 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 86 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Tyr Ile Ile Pro Ser Thr Asp Tyr Thr Glu Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Ile Thr Ala Asp Lys Ser Ala Gly Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Tyr Pro Tyr Arg Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 87 <211> 116 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 87 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Val Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val 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 Asn Pro Gly Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <210> 88 <211> 116 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 88 Gln Ile Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Val Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val 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 Asn Pro Gly Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <210> 89 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 89 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Asn Thr Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 90 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 90 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Asn Thr Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 91 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 91 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 92 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 92 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 93 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 93 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 94 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 94 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 95 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 95 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 96 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain variable region <400> 96 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 97 <211> 107 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain variable region <400> 97 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 98 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 98 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 99 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 99 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 100 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 100 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Phe Trp Asn Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 101 <211> 106 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 101 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 Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Leu Thr Ser Asn Leu Ala Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Thr Asn Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 102 <211> 106 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 102 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 Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Leu Ile Tyr 35 40 45 Leu Thr Ser Asn Leu Ala Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu 65 70 75 80 Asp Ala Ala Val Tyr Tyr Cys Gln Gln Trp Ser Thr Asn Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 103 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 103 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly 1 5 10 15 Gln Arg Ala Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asn Asp Thr Ala Asn Tyr Tyr Cys Gln Gln Ser His 85 90 95 Glu Ala Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 104 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 104 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly 1 5 10 15 Gln Arg Ala Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asn Asp Thr Ala Asn Tyr Tyr Cys Gln Gln Ser His 85 90 95 Glu Ala Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 105 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain variable region <400> 105 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly 1 5 10 15 Gln Arg Ala Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Asn Thr Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Gly Asp Thr Ala Asn Tyr Tyr Cys Gln Gln Ser His 85 90 95 Glu Ala Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 106 <211> 330 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain constant region <400> 106 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Leu 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 Arg 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 Leu 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 Glu Glu 225 230 235 240 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Leu Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 107 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain constant region <400> 107 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 108 <211> 357 <212> DNA <213> Artificial <220> <221> gene <222> ()..() <223> Heavy chain variable region <400> 108 caggtgcagc tgaagcagtc aggacctggc ctagtgcagc cctcacagag cctgtccata 60 acctgcacag tctctggttt ctcattaact agctatggtg tacactgggt tcgccagtct 120 ccaggaaagg gtctggagtg gctggggagtg atatggagag gtggaagcac agactacaat 180 gcagctttca tgtccagact gagcatcacc aaggacaact ccaagagcca agttttcttt 240 aaaatgaaca gtctgcaagc tgatgacact gccatatact actgtgccaa aggtatgatt 300 acgacgggct atgctatgga ctactggggt caaggaacct cagtcaccgt ctcctca 357 <210> 109 <211> 318 <212> DNA <213> Artificial <220> <221> gene <222> ()..() <223> Light chain variable region <400> 109 gacatccaga tgacacaatc ttcatcctcc ttttctgtat ctctaggaga cagagtcacc 60 attacttgca aggcaagtga ggacatatat aatcggttag cctggtatca gcagaaacca 120 ggaaatgctc ctaggctctt aatatctggt gcaaccagtt tggaaactgg ggttccttca 180 agattcagtg gcagtggatc tggaaaggat tacactctca gcattaccag tcttcagact 240 gaagatgttg ctacttatta ctgtcaacag tattggagta ctcccacgtt cggagggggg 300 accaagctgg aaataaaa 318 <210> 110 <211> 357 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> heavy chain variable region <400> 110 caggtgcagc tgaagcagtc aggacctggc ctagtgcagc cctcacagag cctgtccata 60 acctgcacag tctctggttt ctcattaact agctttggta tacactgggt tcgccagtct 120 ccaggaaagg gtctggagtg gctgggagtg atatggagag gtggaagcac agactacaat 180 gcagctttcc tgtccagact gagcatcacc aaggacaact ccaagagcca agttttcttt 240 aaaataaaca gtcttcgagc tgatgacact gccatatact tctgtgccaa atctatgatt 300 acgaccggct atgctatgga ctactggggt caaggaacct cagtcaccgt ctcctca 357 <210> 111 <211> 321 <212> DNA <213> Artificial <220> <221> gene <222> ()..() <223> Light chain variable region <400> 111 gacatccaga tgacacaatc ttcatcctcc ttttctgtat ctctaggaga cagagtcacc 60 attacttgca aggcaagtga ggacatatat aatcggttag cctggtatca gcagaaacca 120 ggaaatgctc ctaggctctt aatatctggt gcaaccagtt tggaaactgg ggttccttca 180 agattcagtg gcagtggatc tggaaaggat tacattctca gcattaccag tcttcagact 240 gaagatgttg ctacttatta ctgtcaacag tattggagta ctccgtggac gttcggtgga 300 ggcaccaagc tggagatcaa a 321 <210> 112 <211> 357 <212> DNA <213> Artificial <220> <221> gene <222> ()..() <223> Heavy chain variable region <400> 112 caggtgcagc tgaagcagtc aggacctggc ctagtgcagc cctcacagag cctgtccata 60 acctgcacag tctctggttt ctcattaact aactatggtg tacactggct tcgccagtct 120 ccaggaaagg gtctggagtg gctgggagtg atatggagag gtggaagcac agactacagt 180 gcagctttca tgtccagact gaacatcacc aaggacaact ccaagagtca agtttttttt 240 aaaatgaaca gtctgcaagc tgatgacact gccatatact actgtgccaa aactacgctt 300 acgagggtct ggtattgcga tgtctggggc gcagggacca cggtcaccgt ctcctca 357 <210> 113 <211> 321 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> Light chain variable region <400> 113 gacatccaga tgacacaatc ttcatcctcc ttttctgtat ctctaggaga cagagtcacc 60 attacttgca agtcaagtga ggacatatat aatcggttag cctggtatca gcagaaacca 120 ggaaatgctc ctaggctctt aatatctggt gcaaccagtt tggaaactgg ggttccttca 180 agattcagtg gcagtggatc tggaaaggat tacactctca gcattaccag tcttcagact 240 gaagatgttg ttacttatta ctgtcaacag tattggaata ctccgctcac gttcggtgct 300 gggaccaagc tggagctgaa a 321 <210> 114 <211> 357 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> heavy chain variable region <400> 114 caggtgcagc tgaagcagtc aggacctggc ctagtgcagc cctcacagag cctgtccata 60 acctgcacag tctctgggtt ctcattactt agttatggtg tacactgggt tcgccagtct 120 ccaggaaagg gtctggagtg gctgggagtg atatggagag gtggaagcgc agactacaat 180 gcagctttca tgtccagact gagcatcacc aaagacaact ccaagagcca agttttcttt 240 aaaatgaaca gtctgcaagc tgatgacact gccatatact actgtgccaa aagtatgatt 300 acgacggggt ttactctgga ctactggggt caaggaacct cagtcaccgt ctcctca 357 <210> 115 <211> 321 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> Light chain variable region <400> 115 gacatccaga tgacacaatc ttcatcctcc ttttctgtat ctctaggaga cagagtcacc 60 attacttgca aggcaagtga ggacatatat aatcggttag cctggtatca gcagaaacca 120 ggacatgctc ctaggctctt aatatctggt gcaaccagtt tggaaactgg ggttccttca 180 agattcagtg gcagtggatc tggaaaggat tacactctca gcattaccag tcttcagact 240 gaagatgttg ctacttatta ctgtcaacag ttttggaata ctccgtggac gttcggtgga 300 ggcaccaagc tggaaatcaa a 321 <210> 116 <211> 342 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> Heavy chain variable region <400> 116 cagatccagt tggtgcagtc tggacctgag ctgaagaagc ctggagagac agtcaagatc 60 tcctgcaagg cttctggata taccttcaca aactttggaa tgaactgggt gaagcaggct 120 ccaggaaagg gtttaaagtg gatgggctgg ataaacacct acactggaga gccaacaaat 180 gctgatgact tcaagggacg gtttgccttc tctttggaaa cctctgccag cactgcctat 240 ttgcagatca acaacctcaa aaatgaggac atggctacat atttctgtgc aagaaaaggg 300 tttgcttact ggggccaagg gactctggtc actgtctctg ca 342 <210> 117 <211> 333 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> light chain variable region <400> 117 gacattgtgc tgacccaatc tccagcttct ttggctgtgt ctctagggca gaggaccacc 60 atatcctgca gagccagtga aagtgttgat agttatggca atagttttat gcactggtac 120 cagcagaaac caggacagcc acccaaactc ctcatctatc gtgcatccaa cctagaatct 180 gggatccctg ccaggttcag tggcagtggg tctaggacag acttcaccct caccattaat 240 cctgtggagg ctgatgatgt tgcaacctat tactgtcagc aaattaatga ggatccattc 300 acgttcggct cggggacaaa gttggaaata aaa 333 <210> 118 <211> 342 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> heavy chain variable region <400> 118 cagatccagt tggtgcagtc tggacctgag ctgaagaagc ctggagagac agtcaagatc 60 tcctgcaagg cttctggata taccttcaca aactatggaa tgaactgggt gaagcaggct 120 ccaggaaagg gtttaaagtg gatgggctgg ataaacacct acactgcaga gccaacatat 180 gctgatgact tcaagggacg gtttgccttc tctttggaag cctctgccag cactgcctat 240 ttgcagatca acaacctcaa aaatgaggac atggctacat atttctgtgc aagaaagtat 300 ggtggctact ggggccaagg caccactctc acagtctcct ca 342 <210> 119 <211> 333 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> light chain variable region <400> 119 gacattgtgc tgacccaatc tccaccttct ttggctgtgt ctctagggca gagggccacc 60 atatcctgca gagccagtga aagtgttgat agttatggca atagttttat acactggtac 120 cagcagaaac caggacagtc acccaaactc ctcatctatc gtgcatccaa cctagaatct 180 gggatccctg ccaggttcag tggcagtggg tctaggacag acttcaccct caccattaat 240 cctgtggagg ctgatgatgt tgcaacctat tactgtcagc aagttaatga ggatccgctc 300 acgttcggtg ctgggaccaa gctggagctg aaa 333 <210> 120 <211> 357 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> Heavy chain variable region <400> 120 caggtccagc ttcagcagtc tggggctgaa ctggcaaaac ctggggcctc agtgaagatg 60 tcctgcaagg cttctggcta cacctttact aactactgga tgaattggat aaaacagagg 120 cctggacagg gtctggaatg gattggatac attattccaa gcactgatta tactgagtac 180 aatcagaagt tcaaggacaa ggccacattg actgcagaca aatcctccgg cacagcctac 240 atgcaactga gcagcctgac atctgaggac tctgcagtct attactgttc aaggtaccct 300 tataggggct atgctatgga ctactggggt caaggaacct cagtcaccgt ctcctca 357 <210> 121 <211> 318 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> light chain variable region <400> 121 caaattgttc tcacccagtc tccagcactc atgtctgcat ctccagggga gaaggtcacc 60 atgacctgca gtgccagctc aagtgtaagt tacatgtact ggtaccaaca gaagccaaga 120 tcctccccca aaccctggat ttatctcaca tccaacctgg cttctggagt ccctgctcgc 180 ttcagtggca gtgggtctgg gacctcttac tctctcacaa tcagcagcat ggaggctgaa 240 gatgctgcca cttattactg ccagcagtgg agtactaacc cgtggacgtt cggtggaggc 300 accaagctgg aactcaaa 318 <210> 122 <211> 348 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> heavy chain variable region <400> 122 cagatccagt tggtgcagtc tggacctgag ctgaagaagc ctggagagac agtcaagatc 60 tcctgcaagg cttctgggta taccttcaca aactatggaa tgaactgggt gaagcaggct 120 ccaggaaagg gtttaaagtg gatgggctgg ataaacacct acactggaga gccaacagtt 180 gctgatgact tcaagggacg gtttgccttc tctttggaca cctctgccag cactgcctat 240 ttgcagatca acaacctcaa aaatgaggac acggctacat atttctgtgc aagaaatccc 300 ggctggtttg cttactgggg ccaagggact ctggtcactg tctctgca 348 <210> 123 <211> 333 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> light chain variable region <400> 123 gacattgtgt tgacccaatc tccagttttt ttggctgtgt ctctagggca gagggccacc 60 atatcctgca gagccagtga aagtgttgat aattatggca atagttttat gcactggtac 120 cagcagaaac caggacagcc acccaaactc ctcatctatc gtgcatccaa cctagaatct 180 gggatccctg ccaggttcag tggcagtggg tctaggacag acttcaccct caccattaat 240 cctgtggagg ctgatgatgt tgcaacctat tactgtcagc aaagtcatga ggctccgtgg 300 acgttcggtg gaggcaccag gctggaaatc aaa 333 <210> 124 <211> 348 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> heavy chain variable region <400> 124 cagatccagt tggtgcagtc tggacctgag ctgaagaagc ctggagagac agtcaagatc 60 tcctgcaagg cttctgggta taacctcaca aactatggaa tgaactgggt gaagcaggct 120 ccaggaaagg gtttaaagtg gatgggctgg ataaacacct acactggaga gccaacatat 180 gctgatgact tcaagggacg gtttgccttc tctttggaaa cctctgccag cactgcctat 240 ttgcagatca acaacctcaa aaatgaggac acggctacat atttctgtgc aagataccca 300 ggctggtttg cttactgggg ccaagggact ctggtcactg tctctgca 348 <210> 125 <211> 333 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> Light chain variable region <400> 125 gacattgtgc tgacccaatc tccagcttct ttggctgtgt ctctagggca gagggccacc 60 atatcctgca gaaccagtga aagtgttgat agttatggca atagttttat gcactggtac 120 cagcagaaac caggacagcc acccaaactc ctcatctatc gtgcatccaa cctagaatct 180 gggatccctg ccaggttcag tggcagtggg tctaggacag acttcaccct caccattaat 240 cctgtggagg ctgatgatgt tgcaacctat tactgtcagc aaagtaatga ggatccgtgg 300 acgttcggtg gaggcaccaa gctggaaatc aaa 333 <210> 126 <211> 990 <212> DNA <213> Artificial <220> <221> gene <222> ()..() <223> Heavy chain constant region <400> 126 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 60 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 120 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 180 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 240 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagag agttgagccc 300 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga 360 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 420 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 480 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 540 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 600 gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc 660 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggaggag 720 atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 780 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 840 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 900 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 960 cagaagagcc tctccctgtc tccgggtaaa 990 <210> 127 <211> 321 <212> DNA <213> artificial <220> <221> gene <222> ()..() <223> light chain constant region <400> 127 cgtacggtgg ctgcaccatc tgtcttcatc ttcccgccat ctgatgagca gttgaaatct 60 ggaactgcct ctgttgtgtg cctgctgaat aacttctatc cccgcgaggc caaagtacag 120 tggaaggtgg ataacgccct ccaatcgggt aactcccagg agagtgtcac agagcaggac 180 agcaaggaca gcacctacag cctcagcagc accctgacgc tgagcaaagc agactacgag 240 aaacacaaag tctacgcctg cgaagtcacc catcagggcc tgagctcgcc cgtcacaaag 300 agcttcaacc gcggagagtg t 321 <210> 128 <211> 122 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Daratumumab, heavy chain variable region <400> 128 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Asn Ser Phe 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Gly Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Lys Asp Lys Ile Leu Trp Phe Gly Glu Pro Val Phe Asp Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 129 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Daratumumab, light chain variable region <400> 129 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 Gln Ser Val Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 130 <211> 120 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Isatuximab, heavy chain variable region <400> 130 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Ala Lys Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Trp Met Gln Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Thr Ile Tyr Pro Gly Asp Gly Asp Thr Gly Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Lys Thr Val Tyr 65 70 75 80 Met His Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Tyr Gly Ser Asn Ser Leu Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 131 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Isatuximab, light chain variable region <400> 131 Asp Ile Val Met Thr Gln Ser His Leu Ser Met Ser Thr Ser Leu Gly 1 5 10 15 Asp Pro Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Val 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Arg Arg Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Ile Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ala Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 132 <211> 120 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Mor202, heavy chain variable region <400> 132 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Gly Asp Pro Ser Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Leu Pro Leu Val Tyr Thr Gly Phe Ala Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 133 <211> 107 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Mor202, light chain variable region <400> 133 Asp Ile Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln 1 5 10 15 Thr Ala Arg Ile Ser Cys Ser Gly Asp Asn Leu Arg His Tyr Tyr Val 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Gly Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Thr Tyr Thr Gly Gly Ala Ser Leu 85 90 95 Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 <210> 134 <211> 106 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 134 Gln Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40 45 Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Thr Asn Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 135 <211> 106 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 135 Gln Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Arg Pro Trp Ile Tyr 35 40 45 Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Thr Asn Pro Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 136 <211> 12 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 136 Ser Ala Ser Ser Ser Val Ser Tyr Met Tyr Trp Tyr 1 5 10 <210> 137 <211> 9 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR3 <400> 137 Ser Met Ile Thr Thr Gly Phe Thr Leu 1 5 <210> 138 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 138 Gln Gln Phe Trp Asn Thr Pro Trp Thr Phe 1 5 10 <210> 139 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR3 <400> 139 Gln Gln Trp Ser Thr Asn Pro Trp Thr Phe 1 5 10 <210> 140 <211> 116 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain variable region <400> 140 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Asn Leu Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val 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 Tyr Pro Gly Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <210> 141 <211> 116 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 141 Gln Ile Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Asn Leu Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val 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 Tyr Pro Gly Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <210> 142 <211> 116 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> heavy chain variable region <400> 142 Gln Ile Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Leu Thr Asn Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val 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 Tyr Pro Gly Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <210> 143 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 143 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly 1 5 10 15 Gln Arg Ala Thr Ile Thr Cys Arg Thr Ser Glu Ser Val Asp Ser Tyr 20 25 30 Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asn Asp Thr Ala Asn Tyr Tyr Cys Gln Gln Ser Asn 85 90 95 Glu Asp Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 144 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 144 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly 1 5 10 15 Gln Arg Ala Thr Ile Thr Cys Arg Thr Ser Glu Ser Val Asp Ser Tyr 20 25 30 Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asn Asp Thr Ala Asn Tyr Tyr Cys Gln Gln Ser Asn 85 90 95 Glu Asp Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 145 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 145 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly 1 5 10 15 Gln Arg Ala Thr Ile Thr Cys Arg Thr Ser Glu Ser Val Asp Ser Tyr 20 25 30 Gly Asn Thr Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Asn Asp Thr Ala Asn Tyr Tyr Cys Gln Gln Ser Asn 85 90 95 Glu Asp Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 146 <211> 111 <212> PRT <213> artificial <220> <221> PEPTIDE <222> ()..() <223> light chain variable region <400> 146 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly 1 5 10 15 Gln Arg Ala Thr Ile Thr Cys Arg Thr Ser Glu Ser Val Asp Ser Tyr 20 25 30 Gly Asn Thr Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Ala Gln Asp Thr Ala Asn Tyr Tyr Cys Gln Gln Ser Asn 85 90 95 Glu Asp Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 147 <211> 10 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Heavy chain, CDR1 <400> 147 Gly Tyr Ala Leu Thr Asn Tyr Gly Met Asn 1 5 10 <210> 148 <211> 15 <212> PRT <213> Artificial <220> <221> PEPTIDE <222> ()..() <223> Light chain, CDR1 <400> 148 Arg Thr Ser Glu Ser Val Asp Ser Tyr Gly Asn Thr Phe Met His 1 5 10 15

Claims

1. An antibody molecule or an antigen-binding fragment thereof that binds to human CD38, wherein the antibody molecule or the antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region and the light chain variable region comprise the following CDR combinations: VH-CDR1 as shown in SEQ ID NO: 16, VH-CDR2 as shown in SEQ ID NO: 17, VH-CDR3 as shown in SEQ ID NO: 18; VL-CDR1 as shown in SEQ ID NO: 4, VL-CDR2 as shown in SEQ ID NO: 5, VL-CDR3 as shown in SEQ ID NO:

19.

2. The antibody molecule or antigen-binding fragment thereof according to claim 1, characterized in that: The heavy chain variable region and light chain variable region comprised by the antibody molecule or antigen-binding fragment thereof are selected from the following combinations: (1) the amino acid sequence shown in SEQ ID NO: 56; and, the amino acid sequence shown in SEQ ID NO: 57; (2) the amino acid sequence shown in SEQ ID NO: 74; and, the amino acid sequence shown in SEQ ID NO: 92; (3) the amino acid sequence shown in SEQ ID NO: 75; and, the amino acid sequence shown in SEQ ID NO: 94; (4) the amino acid sequence shown in SEQ ID NO: 76; and, the amino acid sequence shown in SEQ ID NO: 94; or (5) the amino acid sequence shown in SEQ ID NO: 77; and, the amino acid sequence shown in SEQ ID NO:

95.

3. The antibody molecule or antigen-binding fragment thereof according to claim 1, characterized in that: The antibody molecule or antigen-binding fragment thereof is a monoclonal antibody, a single-chain antibody, a fully or partially humanized antibody or a chimeric antibody.

4. The antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 3, characterized in that: The antibody molecule or antigen-binding fragment thereof is scFv, BsFv, dsFv, (dsFv)2, Fab, Fab', F(ab')2 or Fv.

5. The antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 3, characterized in that: The antibody molecule or antigen-binding fragment thereof further comprises a human or mouse constant region.

6. The antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 3, characterized in that: The antibody molecule or antigen-binding fragment thereof further comprises a human or mouse light chain constant region (CL) and / or a heavy chain constant region (CH).

7. The antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 3, characterized in that: The antibody molecule or antigen-binding fragment thereof comprises a heavy chain constant region selected from IgG, IgA, IgM, IgD or IgE and / or a kappa or lambda type light chain constant region.

8. The antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 3, characterized in that: The antibody molecule is a monoclonal antibody.

9. The antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 3, characterized in that: The antibody molecule is a mouse, chimeric or humanized monoclonal antibody.

10. The antibody molecule or antigen-binding fragment thereof according to claim 9, characterized in that: The heavy chain constant region of the monoclonal antibody is of IgG1 or IgG4 subtype, and the light chain constant region is of κ type.

11. The antibody molecule or antigen-binding fragment thereof according to claim 9, characterized in that: The heavy chain constant region of the monoclonal antibody comprises the amino acid sequence shown in SEQ ID NO:

106.

12. The antibody molecule or antigen-binding fragment thereof according to claim 9, characterized in that: The light chain constant region of the monoclonal antibody comprises the amino acid sequence shown in SEQ ID NO:

107.

13. A nucleic acid molecule encoding the antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 12. A vector comprising the nucleic acid molecule according to claim 13 . 15 . A host cell, comprising the nucleic acid molecule of claim 13 or the vector of claim 14 , or the host cell is transformed or transfected with the nucleic acid molecule of claim 13 or the vector of claim 14 .

16. A pharmaceutical composition comprising the antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 12, the nucleic acid molecule according to claim 13, the vector according to claim 14 or the host cell according to claim 15, and optionally a pharmaceutically acceptable excipient.

17. Use of the antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 12, the nucleic acid molecule according to claim 13, the vector according to claim 14, the host cell according to claim 15 or the pharmaceutical composition according to claim 16 in the preparation of a medicament for treating multiple myeloma or non-Hodgkin's lymphoma.

18. A kit comprising the antibody molecule or antigen-binding fragment thereof according to any one of claims 1 to 12, the nucleic acid molecule according to claim 13, the vector according to claim 14, the host cell according to claim 15 or the pharmaceutical composition according to claim 16.

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