Anti-CD40 antibody and its use

Abstract

The present invention relates to an antibody or an antigen-binding fragment thereof that specifically binds to CD40, and a composition containing the antibody or the antigen-binding fragment. Furthermore, the present invention relates to a nucleic acid encoding the antibody or the antigen-binding fragment, a host cell containing the same, and uses related thereto. In addition, the present invention relates to the therapeutic use and the detection use of the antibody or the antigen-binding fragment.

Description

【Technical Field】 【0001】 The present application relates to an antibody or an antigen-binding fragment thereof that specifically binds to CD40, and a composition containing the antibody or the antigen-binding fragment. Furthermore, the present application relates to a nucleic acid molecule encoding the antibody or the antigen-binding fragment, a host cell containing the nucleic acid molecule, and related uses thereof. Furthermore, the present application relates to the therapeutic use and the detection use of the antibody or the antigen-binding fragment. 【Background Art】 【0002】 CD40, also known as TNFRSF5, belongs to the tumor necrosis factor receptor superfamily (TNFRSF) and is widely expressed in a variety of cell types including immune B cells, antigen-presenting cells (APCs), and some tumor cells. The CD40 precursor contains 297 amino acids (aa) and is a type I transmembrane glycoprotein consisting of an N-terminal signal peptide (20 aa), an extracellular domain (193 aa), a transmembrane domain (22 aa), and a cytoplasmic domain (62 aa). The extracellular domain contains four cysteine-rich domains (CRDs) and has a total of 22 cysteines. CD40 is highly glycosylated, and the molecular weight calculated from the amino acid sequence is 28 kD, but the glycosylated molecular weight is 40 - 50 kD. 【0003】 The ligand of CD40, CD154 (also known as TNFSF5 or CD40L), is a type II membrane integral protein of 34 - 39 kDa. The expression of CD40L is usually inducible and is limited to hematopoietic cells such as platelets, granulocytes, activated T cells, activated B cells, and activated natural killer (NK) cells, but is also slightly expressed on endothelial cells and smooth muscle cells. 【0004】 Large-scale studies have confirmed that CD8+ cytotoxic T lymphocytes are generated by the CD40 - CD40L interaction. This is important in the immune response and is essential for the adaptive immune response. 【0005】 CD40 expression on monocytes, their progeny macrophages and DCs, and B cells plays an important role in immune cell function. Monocytes are innate immune progenitor cells with very high flexibility. Monocytes have the ability to differentiate into several cell types, such as myeloid-derived suppressor cells (MDSC), macrophages, and DCs. CD40 signaling is an important trigger for the monocyte maturation process and mainly drives the differentiation into M1-spectrum macrophages and DCs. The involvement of CD40 on the DC surface promotes the production of cytokines and chemokines, induces the expression of co-stimulatory molecules, and enhances antigen cross-presentation. One of the main functions of CD40L is to enhance antigen presentation to T cells by activating DCs. This step, called "licensing", activates the latter by increasing the interaction between DCs and T cells through upregulation of surface proteins such as CD54 and CD86. 【0006】 B cells are likewise targets of CD40L activity. The interaction between B cells and activated T cells expressing CD40L also increases the expression of MHC-II and co-stimulatory molecules, such as CD80 or CD86, and induces Ig class switching in B cells. Activated B cells migrate to lymphoid organs that present antigens to T cells, and CD40-activated DCs and B cells support the immune response by releasing immunostimulatory cytokines and chemokines, such as IL-12p70, CXCL10, and TNFα. In addition, CD40-activated B cells can stimulate antigen-specific CD8+ T cells by enhancing the secretion of cytokines, including TNFα and IFNγ. Several studies have provided evidence that CD40-expressing B cells activated ex vivo become fully functional antigen-presenting B cells, and subsequent adoptive cell transfer (ACT) therapy with such cells improves anti-tumor activity. 【0007】 CD40 plays a central role in the anti-tumor immune response, and various strategies to induce CD40 signaling have been extensively studied and explored. Current drug development targeting CD40 can be broadly classified into agonists (activators) or antagonists (inhibitors) depending on whether the goal is to enhance immune activity in the tumor microenvironment or to suppress the immune system in autoimmune diseases. However, as a monotherapy, CD40 antibodies have only demonstrated moderate activity, and the tumor objective response rate (ORR) was less than 20%. This is particularly the case in "cold tumors" with insufficient immune responses, i.e., tumors where immune cells have not yet extensively infiltrated. As a result, many CD40 monoclonal antibody projects have been discontinued due to such limitations. 【0008】 Considering the significance of CD40 in cancer immunity, CD40 is considered an extremely attractive target for cancer immunotherapy. There is a strong need in the art for the development of new anti-CD40 antibodies, especially anti-CD40 antibodies that can target various epitopes and can block or not block CD40L ligand binding. Such antibodies have various applicability in tumor immunity and autoimmune diseases and can be used for the treatment of diseases, especially cancer therapy, when combined with clinical combination therapies. 【Summary of the Invention】 【Problems to be Solved by the Invention】 【0009】 After extensive research, the inventors of the present application screened and obtained a series of fully human antibodies against CD40 that have high binding affinity for CD40, show cross-reactivity with human CD40 and cynomolgus monkey CD40, and can efficiently block or not block the binding to the ligand CD40L of CD40. Based on this, the present application also provides a composition containing an antibody or an antigen-binding fragment thereof, a nucleic acid molecule encoding the antibody or the antigen-binding fragment thereof, a host cell containing the same, and related uses. 【Means for Solving the Problems】 【0010】 Antibody or antigen-binding fragment Thus, in one aspect, an antibody or antigen-binding fragment thereof that can specifically bind to CD40, (1) The following three complementarity-determining regions (CDRs) of the heavy-chain variable region (VH): (a) VH CDR1 having a structure selected from the group consisting of FTFX1X2YX3MH (SEQ ID NO: 76), GSISSYYWS (SEQ ID NO: 6) (b) VH CDR2 having a structure selected from the group consisting of VIX4YX5X6X7X8KYYAX9SVKG (SEQ ID NO: 77), GISWSSX 10 SIX 11 YADSVKG (SEQ ID NO: 78), SIYYSGSTNYNPSLKS (SEQ ID NO: 22) (c) VH CDR3 having a structure selected from the group consisting of ARDTSRGHDI (SEQ ID NO: 23), AKDPTATTGARGYFDL (SEQ ID NO: 24), ARDANYYKWHPY (SEQ ID NO: 25) And / or (2) The following three complementarity-determining regions (CDRs) of the light-chain variable region (VL): (d) VL CDR1 having a structure selected from the group consisting of X 12 ASQSVSX 13 X 14 YLA (SEQ ID NO: 79), X 15 ASX 16 X 17 ISSWLA (SEQ ID NO: 7) (e) VL CDR2 having a structure selected from the group consisting of GASX 18 X 19 X 20 X 21 (SEQ ID NO: 8), AASX 22 LX 23 S (SEQ ID NO: 9) (f) X 24 X 25 YX 26 DYPPFT (SEQ ID NO: 10), QQX 27 X​​​​​28 SX 29 PPT (SEQ ID NO: 11) A VL CDR3 having a structure selected from the group consisting of comprising X1 is selected from the group consisting of (i) the amino acid residues S, E, G, D and (ii) amino acid residues that are conservative substitutions for (i), X2 is selected from the group consisting of (i) the amino acid residues S, K, D, T and (ii) amino acid residues that are conservative substitutions for (i), X3 is selected from the group consisting of (i) the amino acid residues G, A and (ii) amino acid residues that are conservative substitutions for (i), X4 is selected from the group consisting of (i) the amino acid residues S, H and (ii) amino acid residues that are conservative substitutions for (i), X5 is selected from the group consisting of (i) the amino acid residues E, H and (ii) amino acid residues that are conservative substitutions for (i), X6 is selected from the group consisting of (i) the amino acid residues G, A and (ii) amino acid residues that are conservative substitutions for (i), X7 is selected from the group consisting of (i) the amino acid residues S, E, T, V and (ii) amino acid residues that are conservative substitutions for (i), X8 is selected from the group consisting of (i) the amino acid residues N, S, I, K and (ii) amino acid residues that are conservative substitutions for (i), X9 is selected from the group consisting of (i) the amino acid residues D, E and (ii) amino acid residues that are conservative substitutions for (i), X 10 is selected from the group consisting of (i) the amino acid residues G, D and (ii) amino acid residues that are conservative substitutions for (i), X 11 is selected from the group consisting of (i) the amino acid residues G, H and (ii) amino acid residues that are conservative substitutions for (i), X 12 is selected from the group consisting of (i) the amino acid residues R, Q and (ii) amino acid residues that are conservative substitutions for (i), X 13is selected from the group consisting of (i) amino acid residues Y, S and (ii) amino acid residues that are conservative substitutions for (i), X 14 is selected from the group consisting of (i) amino acid residues S, D, N and (ii) amino acid residues that are conservative substitutions for (i), X 15 is selected from the group consisting of (i) amino acid residues R, E and (ii) amino acid residues that are conservative substitutions for (i), X 16 is selected from the group consisting of (i) amino acid residues Q, K and (ii) amino acid residues that are conservative substitutions for (i), X 17 is selected from the group consisting of (i) amino acid residues G, V and (ii) amino acid residues that are conservative substitutions for (i), X 18 is selected from the group consisting of (i) amino acid residues S, T, A and (ii) amino acid residues that are conservative substitutions for (i), X 19 is selected from the group consisting of (i) amino acid residues R, L and (ii) amino acid residues that are conservative substitutions for (i), X 20 is selected from the group consisting of (i) amino acid residues N, A and (ii) amino acid residues that are conservative substitutions for (i), X 21 is selected from the group consisting of (i) amino acid residues T, N and (ii) amino acid residues that are conservative substitutions for (i), X 22 is selected from the group consisting of (i) amino acid residues S, Y and (ii) amino acid residues that are conservative substitutions for (i), X 23 is selected from the group consisting of (i) amino acid residues E, Q and (ii) amino acid residues that are conservative substitutions for (i), X 24 is selected from the group consisting of (i) amino acid residues S, G, Q and (ii) amino acid residues that are conservative substitutions for (i), X 25is selected from the group consisting of (i) amino acid residues E, Q, D and (ii) amino acid residues that are conservative substitutions for (i), X 26 is selected from the group consisting of (i) amino acid residues A, S and (ii) amino acid residues that are conservative substitutions for (i), X 27 is selected from the group consisting of (i) amino acid residues R, H, V and (ii) amino acid residues that are conservative substitutions for (i), X 28 is selected from the group consisting of (i) amino acid residues S, L, A and (ii) amino acid residues that are conservative substitutions for (i), X 29 is selected from the group consisting of (i) amino acid residues F, Y and (ii) amino acid residues that are conservative substitutions for (i), The antibody or antigen-binding fragment thereof is provided by the present application. 【0011】 (ADI-47754 family) In certain embodiments, the antibody or antigen-binding fragment thereof is (1) The following three complementarity-determining regions (CDRs) of the heavy-chain variable region (VH): (a) VH CDR1 having the structure shown in FTFX1X2YX3MH (SEQ ID NO: 76), (b) VH CDR2 having the structure shown in VIX4YX5X6X7X8KYYAX9SVKG (SEQ ID NO: 77), (c) VH CDR3 having the structure shown in ARDTSRGHDI (SEQ ID NO: 23) and / or (2) The following three complementarity-determining regions (CDRs) of the light-chain variable region (VL): (d) X 12 ASQSVSX 13 X 14 VL CDR1 having the structure shown in YLA (SEQ ID NO: 79), (e) GASX 18 X 19 X 20 X 21 (SEQ ID NO: 8) having the structure shown in VL CDR2, (f)X 24 X 25 YX 26 VL CDR3 having the structure shown in DYPPFT (SEQ ID NO: 10) comprising X1 is selected from the group consisting of (i) amino acid residues S, E, G and (ii) amino acid residues that are conservative substitutions for (i); X2 is selected from the group consisting of (i) amino acid residues S, K and (ii) amino acid residues that are conservative substitutions for (i); X3 is selected from the group consisting of (i) amino acid residue G and (ii) amino acid residues that are conservative substitutions for (i); X4 is selected from the group consisting of (i) amino acid residues S, H and (ii) amino acid residues that are conservative substitutions for (i); X5 is selected from the group consisting of (i) amino acid residues E, H and (ii) amino acid residues that are conservative substitutions for (i); X6 is selected from the group consisting of (i) amino acid residues G, A and (ii) amino acid residues that are conservative substitutions for (i); X7 is selected from the group consisting of (i) amino acid residues S, E, T, V and (ii) amino acid residues that are conservative substitutions for (i); X8 is selected from the group consisting of (i) amino acid residues N, S, I, K and (ii) amino acid residues that are conservative substitutions for (i); X9 is selected from the group consisting of (i) amino acid residues D, E and (ii) amino acid residues that are conservative substitutions for (i); X 12 is selected from the group consisting of (i) amino acid residues R, Q and (ii) amino acid residues that are conservative substitutions for (i); X 13 is selected from the group consisting of (i) amino acid residues Y, S and (ii) amino acid residues that are conservative substitutions for (i); X 14 is selected from the group consisting of (i) amino acid residues S, D, N and (ii) amino acid residues that are conservative substitutions for (i); X 18is selected from the group consisting of (i) amino acid residues S, T, A and (ii) amino acid residues that are conservative substitutions for (i), X 19 is selected from the group consisting of (i) amino acid residues R, L and (ii) amino acid residues that are conservative substitutions for (i), X 20 is selected from the group consisting of (i) amino acid residues N, A and (ii) amino acid residues that are conservative substitutions for (i), X 21 is selected from the group consisting of (i) amino acid residues T, N and (ii) amino acid residues that are conservative substitutions for (i), X 24 is selected from the group consisting of (i) amino acid residues S, G, Q and (ii) amino acid residues that are conservative substitutions for (i), X 25 is selected from the group consisting of (i) amino acid residues E, Q, D and (ii) amino acid residues that are conservative substitutions for (i), X 26 is selected from the group consisting of (i) amino acid residues A, S and (ii) amino acid residues that are conservative substitutions for (i). 【0012】 In certain embodiments, X1 is selected from the group consisting of amino acid residues S, E, G; X2 is selected from the group consisting of amino acid residues S, K; X3 is selected from the group consisting of amino acid residues G; X4 is selected from the group consisting of amino acid residues S, H; X5 is selected from the group consisting of amino acid residues E, H; X6 is selected from the group consisting of amino acid residues G, A; X7 is selected from the group consisting of amino acid residues S, E, T, V; X8 is selected from the group consisting of amino acid residues N, S, I, K; X9 is selected from the group consisting of amino acid residues D, E; X 12 is selected from the group consisting of amino acid residues R, Q; X 13 is selected from the group consisting of amino acid residues Y, S; X 14 is selected from the group consisting of amino acid residues S, D, N; X 18 is selected from the group consisting of amino acid residues S, T, A; X 19is selected from the group consisting of amino acid residues R and L, and X 20 is selected from the group consisting of amino acid residues N and A, and X 21 is selected from the group consisting of amino acid residues T and N, and X 24 is selected from the group consisting of amino acid residues S, G, and Q, and X 25 is selected from the group consisting of amino acid residues E, Q, and D, and X 26 is selected from the group consisting of amino acid residues A and S. 【0013】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises VH CDR1 shown in any one of SEQ ID NOs: 1 to 3, VH CDR2 shown in any one of SEQ ID NOs: 12 to 18, 82, and VH CDR3 shown in SEQ ID NO: 23, VL CDR1 shown in any one of SEQ ID NOs: 37 to 40, VL CDR2 shown in any one of SEQ ID NOs: 45 to 50, and VL CDR3 shown in any one of SEQ ID NOs: 54 to 57 is included. 【0014】 In certain embodiments, the antibody or antigen-binding fragment thereof (1) VH CDR1 shown in SEQ ID NO: 1, VH CDR2 shown in SEQ ID NO: 12, and VH CDR3 shown in SEQ ID NO: 23, VL CDR1 shown in SEQ ID NO: 37, VL CDR2 shown in SEQ ID NO: 45, and VL CDR3 shown in SEQ ID NO: 54, (2) VH CDR1 shown in SEQ ID NO: 1, VH CDR2 shown in SEQ ID NO: 13, and VH CDR3 shown in SEQ ID NO: 23, VL CDR1 shown in SEQ ID NO: 37, VL CDR2 shown in SEQ ID NO: 45, and VL CDR3 shown in SEQ ID NO: 54, (3) VH CDR1 shown in SEQ ID NO: 2, VH CDR2 shown in SEQ ID NO: 14, and VH CDR3 shown in SEQ ID NO: 23, VL CDR1 shown in SEQ ID NO: 38, VL CDR2 shown in SEQ ID NO: 46, and VL CDR3 shown in SEQ ID NO: 55, (4) The VH CDR1 shown in SEQ ID NO: 2, the VH CDR2 shown in SEQ ID NO: 15, and the VH CDR3 shown in SEQ ID NO: 23, the VL CDR1 shown in SEQ ID NO: 39, the VL CDR2 shown in SEQ ID NO: 47, and the VL CDR3 shown in SEQ ID NO: 56 (5) The VH CDR1 shown in SEQ ID NO: 2, the VH CDR2 shown in SEQ ID NO: 16, and the VH CDR3 shown in SEQ ID NO: 23, the VL CDR1 shown in SEQ ID NO: 39, the VL CDR2 shown in SEQ ID NO: 48, and the VL CDR3 shown in SEQ ID NO: 57 (6) The VH CDR1 shown in SEQ ID NO: 2, the VH CDR2 shown in SEQ ID NO: 17, and the VH CDR3 shown in SEQ ID NO: 23, the VL CDR1 shown in SEQ ID NO: 40, the VL CDR2 shown in SEQ ID NO: 49, and the VL CDR3 shown in SEQ ID NO: 57 (7) The VH CDR1 shown in SEQ ID NO: 3, the VH CDR2 shown in SEQ ID NO: 18, and the VH CDR3 shown in SEQ ID NO: 23, the VL CDR1 shown in SEQ ID NO: 38, the VL CDR2 shown in SEQ ID NO: 50, and the VL CDR3 shown in SEQ ID NO: 54, or (8) The VH CDR1 shown in SEQ ID NO: 2, the VH CDR2 shown in SEQ ID NO: 82, and the VH CDR3 shown in SEQ ID NO: 23, the VL CDR1 shown in SEQ ID NO: 39, the VL CDR2 shown in SEQ ID NO: 48, and the VL CDR3 shown in SEQ ID NO: 57 comprises. 【0015】 In certain embodiments, the antibody or antigen-binding fragment thereof further comprises a framework region of a human immunoglobulin. For example, the antibody or antigen-binding fragment thereof comprises a framework region contained in an amino acid sequence encoded by a human germline antibody gene. For example, the antibody or antigen-binding fragment thereof comprises a heavy chain framework region contained in an amino acid sequence encoded by a human heavy chain germline gene and / or a light chain framework region contained in an amino acid sequence encoded by a human light chain germline gene. 【0016】 In certain embodiments, the antibody or antigen-binding fragment thereof (1) A heavy chain variable region (VH) comprising an amino acid sequence selected from the following: (i) an array shown in any one of SEQ ID NOs: 26 - 32, 83, (ii) an array having one or several amino acid substitutions, deletions or additions (for example, substitution, deletion or addition of 1, 2, 3, 4 or 5 amino acids) compared with the array shown in any one of SEQ ID NOs: 26 - 32, 83, or (iii) an array having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity compared with the array described in any one of SEQ ID NOs: 26 - 32, 83, and / or (2) a light chain variable region (VL) containing an amino acid sequence selected from the following: (i) an array shown in any one of SEQ ID NOs: 61 - 66, (ii) an array having one or several amino acid substitutions, deletions or additions (for example, substitution, deletion or addition of 1, 2, 3, 4 or 5 amino acids) compared with the array shown in any one of SEQ ID NOs: 61 - 66, or (iii) an array having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity compared with the array described in any one of SEQ ID NOs: 61 - 66 is included. 【0017】 In a particular embodiment, the antibody or antigen - binding fragment thereof comprises a VH containing the sequence shown in any one of SEQ ID NOs: 26 - 32, 83 or a variant thereof and a VL containing the sequence shown in any one of SEQ ID NOs: 61 - 66 or a variant thereof, This variant has one or several amino acid substitutions, deletions or additions (e.g., substitution, deletion or addition of 1, 2, 3, 4 or 5 amino acids) compared to the original sequence, or has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity. In certain embodiments, the substitution is a conservative substitution. 【0018】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence shown in SEQ ID NO: 26 or a variant thereof and a VL having the sequence shown in SEQ ID NO: 61 or a variant thereof. 【0019】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence shown in SEQ ID NO: 27 or a variant thereof and a VL having the sequence shown in SEQ ID NO: 61 or a variant thereof. 【0020】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence shown in SEQ ID NO: 28 or a variant thereof and a VL having the sequence shown in SEQ ID NO: 62 or a variant thereof. 【0021】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence shown in SEQ ID NO: 29 or a variant thereof and a VL having the sequence shown in SEQ ID NO: 63 or a variant thereof. 【0022】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence shown in SEQ ID NO: 30 or a variant thereof and a VL having the sequence shown in SEQ ID NO: 64 or a variant thereof. 【0023】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence shown in SEQ ID NO: 31 or a variant thereof and a VL having the sequence shown in SEQ ID NO: 65 or a variant thereof. 【0024】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 32 or a variant thereof and a VL having the sequence set forth in SEQ ID NO: 66 or a variant thereof. 【0025】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 83 or a variant thereof and a VL having the sequence set forth in SEQ ID NO: 64 or a variant thereof, wherein the variant has one or several amino acid substitutions, deletions or additions (e.g., 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) compared to the originating sequence, or has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity. In certain embodiments, the substitution is a conservative substitution. 【0026】 In certain embodiments, the antibody or antigen-binding fragment thereof is capable of blocking the binding to the ligand CD40L of CD40. 【0027】 (ADI-47720 family) In some embodiments, the antibody or antigen-binding fragment thereof (1) the following three complementarity-determining regions (CDRs) of the heavy chain variable region (VH): (a) VH CDR1 having the structure shown in FTFX1X2YX3MH (SEQ ID NO: 76), (b) GISWSSX 10 SIX 11 (c) VH CDR2 having the structure shown in YADSVKG (SEQ ID NO: 78), (c) VH CDR3 having the structure shown in AKDPTATTGARGYFDL (SEQ ID NO: 24), and / or (2) the following three complementarity-determining regions (CDRs) of the light chain variable region (VL): (d) X 15ASX 16 X 17 A VL CDR1 having the structure shown in ISSWLA (SEQ ID NO: 7), (e) AASX 22 LX 23 A VL CDR2 having the structure shown in S (SEQ ID NO: 9), (f) QQX 27 X 28 SX 29 A VL CDR3 having the structure shown in PPT (SEQ ID NO: 11) comprising wherein X1 is selected from the group consisting of (i) amino acid residue D and (ii) amino acid residues that are conservative substitutions for (i), wherein X2 is selected from the group consisting of (i) amino acid residues D, T and (ii) amino acid residues that are conservative substitutions for (i), wherein X3 is selected from the group consisting of (i) amino acid residue A and (ii) amino acid residues that are conservative substitutions for (i), X 10 is selected from the group consisting of (i) amino acid residues G, D and (ii) amino acid residues that are conservative substitutions for (i), X 11 is selected from the group consisting of (i) amino acid residues G, H and (ii) amino acid residues that are conservative substitutions for (i), X 15 is selected from the group consisting of (i) amino acid residues R, E and (ii) amino acid residues that are conservative substitutions for (i), X 16 is selected from the group consisting of (i) amino acid residues Q, K and (ii) amino acid residues that are conservative substitutions for (i), X 17 is selected from the group consisting of (i) amino acid residues G, V and (ii) amino acid residues that are conservative substitutions for (i), X 22 is selected from the group consisting of (i) amino acid residues S, Y and (ii) amino acid residues that are conservative substitutions for (i), X 23is selected from the group consisting of (i) amino acid residues E, Q and (ii) amino acid residues that are conservative substitutions for (i). X 27 is selected from the group consisting of (i) amino acid residues R, H and (ii) amino acid residues that are conservative substitutions for (i). X 28 is selected from the group consisting of (i) amino acid residues S, A and (ii) amino acid residues that are conservative substitutions for (i). X 29 is selected from the group consisting of (i) amino acid residue F and (ii) amino acid residues that are conservative substitutions for (i). 【0028】 In certain embodiments, X1 is amino acid residue D, X2 is selected from the group consisting of amino acid residues D, T, X3 is amino acid residue A, X 10 is selected from the group consisting of amino acid residues G, D, X 11 is selected from the group consisting of amino acid residues G, H, X 15 is selected from the group consisting of amino acid residues R, E, X 16 is selected from the group consisting of amino acid residues Q, K, X 17 is selected from the group consisting of amino acid residues G, V, X 22 is selected from the group consisting of amino acid residues S, Y, X 23 is selected from the group consisting of amino acid residues E, Q, X 27 is selected from the group consisting of amino acid residues R, H, X 28 is selected from the group consisting of amino acid residues S, A, X 29 is amino acid residue F. 【0029】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises VH CDR1 shown in SEQ ID NO: 4 or 5, VH CDR2 shown in any one of SEQ ID NOs: 19-21, and VH CDR3 shown in SEQ ID NO: 24, VL CDR1 shown in any one of SEQ ID NOs: 41-43, VL CDR2 shown in any one of SEQ ID NOs: 51-53, and VL CDR3 shown in SEQ ID NO: 58 or 59. 【0030】 In certain embodiments, the antibody or antigen-binding fragment thereof (1) VH CDR1 shown in SEQ ID NO: 4, VH CDR2 shown in SEQ ID NO: 19, and VH CDR3 shown in SEQ ID NO: 24, VL CDR1 shown in SEQ ID NO: 41, VL CDR2 shown in SEQ ID NO: 51, and VL CDR3 shown in SEQ ID NO: 58, (2) VH CDR1 shown in SEQ ID NO: 4, VH CDR2 shown in SEQ ID NO: 20, and VH CDR3 shown in SEQ ID NO: 24, VL CDR1 shown in SEQ ID NO: 42, VL CDR2 shown in SEQ ID NO: 52, and VL CDR3 shown in SEQ ID NO: 59, or (3) VH CDR1 shown in SEQ ID NO: 5, VH CDR2 shown in SEQ ID NO: 21, and VH CDR3 shown in SEQ ID NO: 24, VL CDR1 shown in SEQ ID NO: 43, VL CDR2 shown in SEQ ID NO: 53, and VL CDR3 shown in SEQ ID NO: 59 comprises. 【0031】 In certain embodiments, the antibody or antigen-binding fragment thereof further comprises a framework region of a human immunoglobulin. For example, the antibody or antigen-binding fragment thereof comprises a framework region contained in the amino acid sequence encoded by a human germline antibody gene. For example, the antibody or antigen-binding fragment thereof comprises a heavy chain framework region contained in the amino acid sequence encoded by a human heavy chain germline gene and / or a light chain framework region contained in the amino acid sequence encoded by a human light chain germline gene. 【0032】 In certain embodiments, the antibody or antigen-binding fragment thereof (1) comprises a heavy chain variable region (VH) having an amino acid sequence selected from the following: (i) the sequence shown in any one of SEQ ID NOS: 33 to 35, (ii) a sequence having one or several amino acid substitutions, deletions or additions (e.g., 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) as compared to the sequence shown in any one of SEQ ID NOS: 33 to 35, or (iii) An array having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity when compared with the array described in any one of SEQ ID NOs: 33 to 35 and / or (2) A variable light chain region (VL) comprising an amino acid sequence selected from the following: (i) The sequence shown in any one of SEQ ID NOs: 67 to 69 (ii) A sequence having one or several amino acid substitutions, deletions or additions (e.g., substitution, deletion or addition of 1, 2, 3, 4 or 5 amino acids) when compared with the sequence shown in any one of SEQ ID NOs: 67 to 69, or (iii) An array having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity when compared with the array described in any one of SEQ ID NOs: 67 to 69 is included. 【0033】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH that is the sequence shown in any one of SEQ ID NOs: 33 to 35 or a variant thereof and a VL that is the sequence shown in any one of SEQ ID NOs: 67 to 69 or a variant thereof, wherein the variant has one or several amino acid substitutions, deletions or additions (e.g., substitution, deletion or addition of 1, 2, 3, 4 or 5 amino acids) when compared with the derived sequence, or has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity. In certain embodiments, the substitution is a conservative substitution. 【0034】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 33 or a variant thereof and a VL having the sequence set forth in SEQ ID NO: 67 or a variant thereof. 【0035】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 34 or a variant thereof and a VL having the sequence set forth in SEQ ID NO: 68 or a variant thereof. 【0036】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH having the sequence set forth in SEQ ID NO: 35 or a variant thereof and a VL having the sequence set forth in SEQ ID NO: 69 or a variant thereof, wherein the variant has one or several amino acid substitutions, deletions or additions (e.g., 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) compared to the derived sequence, or has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity. In certain embodiments, the substitution is a conservative substitution. 【0037】 In some embodiments, the antibody or antigen-binding fragment thereof does not block the binding to the ligand CD40L of human CD40. 【0038】 (ADI-47741 family) In certain embodiments, the antibody or antigen-binding fragment thereof (1) the following three complementarity-determining regions (CDRs) of the heavy-chain variable region (VH): (a) VH CDR1 having the structure shown in GSISSYYWS (SEQ ID NO: 6), (b) VH CDR2 having the structure shown in SIYYSGSTNYNPSLKS (SEQ ID NO: 22), (c) VH CDR3 having the structure shown in ARDANYYKWHPY (SEQ ID NO: 25), and / or (2) The following three complementarity-determining regions (CDRs) of the light chain variable region (VL): (d) X 12 ASQSVSX 13 X 14 VL CDR1 having the structure shown in YLA (SEQ ID NO: 79), (e) GASX 18 X 19 X 20 X 21 VL CDR2 having the structure shown in (SEQ ID NO: 8), (f) QQX 27 X 28 SX 29 VL CDR3 having the structure shown in PPT (SEQ ID NO: 11) comprising X 12 is selected from the group consisting of (i) amino acid residue R and (ii) amino acid residues that are conservative substitutions for (i), X 13 is selected from the group consisting of (i) amino acid residue S and (ii) amino acid residues that are conservative substitutions for (i), X 14 is selected from the group consisting of (i) amino acid residue D and (ii) amino acid residues that are conservative substitutions for (i), X 18 is selected from the group consisting of (i) amino acid residue S and (ii) amino acid residues that are conservative substitutions for (i), X 19 is selected from the group consisting of (i) amino acid residue R and (ii) amino acid residues that are conservative substitutions for (i), X 20 is selected from the group consisting of (i) amino acid residue A and (ii) amino acid residues that are conservative substitutions for (i), X 21 is selected from the group consisting of (i) amino acid residue T and (ii) amino acid residues that are conservative substitutions for (i), X 27 is selected from the group consisting of (i) amino acid residue V and (ii) amino acid residues that are conservative substitutions for (i), X28 is selected from the group consisting of (i) amino acid residue L and (ii) amino acid residues that are conservative substitutions for (i). X 29 is selected from the group consisting of (i) amino acid residue Y and (ii) amino acid residues that are conservative substitutions for (i). 【0039】 In certain embodiments, X 12 is amino acid residue R, X 13 is amino acid residue S, X 14 is amino acid residue D, X 18 is amino acid residue S, X 19 is amino acid residue R, X 20 is amino acid residue A, X 21 is amino acid residue T, X 27 is amino acid residue V, X 28 is amino acid residue L, X 29 is amino acid residue Y. 【0040】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises VH CDR1 set forth in SEQ ID NO: 6, VH CDR2 set forth in SEQ ID NO: 22, and VH CDR3 set forth in SEQ ID NO: 25, VL CDR1 set forth in SEQ ID NO: 44, VL CDR2 set forth in SEQ ID NO: 45, and VL CDR3 set forth in SEQ ID NO: 60. 【0041】 In certain embodiments, the antibody or antigen-binding fragment thereof further comprises a framework region of a human immunoglobulin. For example, the antibody or antigen-binding fragment thereof comprises a framework region contained in the amino acid sequence encoded by a human germline antibody gene. For example, the antibody or antigen-binding fragment thereof comprises a heavy chain framework region contained in the amino acid sequence encoded by a human heavy chain germline gene and / or a light chain framework region contained in the amino acid sequence encoded by a human light chain germline gene. 【0042】 In certain embodiments, the antibody or antigen-binding fragment thereof (1) A heavy chain variable region (VH) comprising an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 36, (ii) A sequence having one or several amino acid substitutions, deletions or additions (e.g., 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) compared to the sequence shown in SEQ ID NO: 36, or (iii) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity compared to the sequence set forth in SEQ ID NO: 36, and / or (2) A light chain variable region (VL) comprising an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 70, (ii) A sequence having one or several amino acid substitutions, deletions or additions (e.g., 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) compared to the sequence shown in SEQ ID NO: 70, or (iii) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity compared to the sequence set forth in SEQ ID NO: 70 is included. 【0043】 In certain embodiments, the antibody or antigen-binding fragment thereof comprises a VH that is the sequence shown in SEQ ID NO: 36 or a variant thereof and a VL that is the sequence shown in SEQ ID NO: 70 or a variant thereof and This variant has one or several amino acid substitutions, deletions or additions (e.g., substitution, deletion or addition of 1, 2, 3, 4 or 5 amino acids) compared to the derived sequence, or has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% sequence identity. In certain embodiments, the substitution is a conservative substitution. 【0044】 In certain embodiments, the antibody or antigen-binding fragment thereof is capable of blocking the binding to the ligand CD40L of human CD40. 【0045】 In certain embodiments, the antibody or antigen-binding fragment thereof further comprises a constant region derived from human immunoglobulin. 【0046】 In certain embodiments, the heavy chain of the antibody or antigen-binding fragment thereof comprises the heavy chain constant region (CH) of human immunoglobulin or a variant thereof, which variant has one or several amino acid substitutions, deletions or additions (e.g., up to 20, up to 15, up to 10 or up to 5 amino acid substitutions, deletions or additions, e.g., substitution, deletion or addition of 1, 2, 3, 4 or 5 amino acids) compared to the derived sequence, and / or the light chain of the antibody or antigen-binding fragment thereof comprises the light chain constant region (CL) of human immunoglobulin or a variant thereof, which variant has up to 20 conservative amino acid substitutions (e.g., up to 15, up to 10 or up to 5 conservative substitutions, e.g., substitution of 1, 2, 3, 4 or 5 amino acids) compared to the derived sequence. 【0047】 In some embodiments, the variant of the heavy chain constant region (CH) may have one or more conservative amino acid substitutions compared to the derived sequence. In such embodiments, the variant of the heavy chain constant region (CH) may have the same or substantially the same effector function compared to the derived wild-type sequence. 【0048】 In other embodiments, the variant of the heavy chain constant region (CH) contains one or more amino acid mutations or chemical modifications that alter one or more of the following properties of the antibody of the present invention: Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function or complement function. The functional change can be achieved by substituting at least one amino acid residue of the constant region of the antibody with a different residue, or by chemically modifying at least one amino acid residue of the antibody, for example, by changing the affinity for an effector ligand (e.g., FcR or complement C1q) of the antibody to alter effector functions, such as some important effector functions mediated by the Fc region of the antibody including ADCC, phagocytosis, CDC, etc. (e.g., decreasing or enhancing). 【0049】 In certain embodiments, the antibody or antigen-binding fragment thereof has ADCC activity. In certain embodiments, the antibody or antigen-binding fragment thereof contains a mutated (e.g., amino acid substitution) or chemically modified Fc region, and this mutated or chemically modified Fc region enables enhancement of ADCC activity. In certain embodiments, the antibody or antigen-binding fragment thereof is produced by expression in a host cell having low fucosylation activity or no fucosylation activity at all, and the produced antibody has low fucosylation or no fucosylation at all that enhances ADCC activity. Such host cells can be mammalian cells, e.g., CHO cells, that do not contain a gene encoding fucosyltransferase (e.g., FUT8). 【0050】 In certain embodiments, the heavy chain of the antibody or antigen-binding fragment thereof contains a heavy chain constant region derived from a human immunoglobulin (e.g., IgG1, IgG2, IgG3 or IgG4). 【0051】 In certain exemplary embodiments, the heavy chain of the antibody or antigen-binding fragment thereof contains the heavy chain constant region shown in SEQ ID NO: 155. 【0052】 In certain embodiments, the light chain of the antibody or antigen-binding fragment thereof comprises a light chain constant region derived from a human immunoglobulin (e.g., the constant region of the κ or λ chain). 【0053】 In certain exemplary embodiments, the light chain of the antibody or antigen-binding fragment thereof comprises the light chain constant region shown in SEQ ID NO: 156. 【0054】 In certain embodiments, the antibody or antigen-binding fragment thereof is selected from the group consisting of Fab, Fab’, F(ab’)2, Fv, disulfide-bonded Fv, scFv, diabody, and single domain antibody (sdAb), and / or the antibody is a chimeric antibody, bispecific antibody or multispecific antibody. 【0055】 In certain embodiments, the antibody or antigen-binding fragment thereof has cross-reactivity with human CD40 and cynomolgus CD40. 【0056】 The antibodies of the present invention can be prepared by various methods known in the art, such as methods using genetic modification techniques and recombinant techniques. For example, the DNA molecules encoding the heavy and light chains of the antibodies of the present invention can be obtained by chemical synthesis or PCR amplification. The resulting DNA molecules are inserted into an expression vector and then transfected into a host cell. The transfected host cell is then cultured under specific conditions to express the antibodies of the present invention. 【0057】 The antigen-binding fragment of the present invention can be obtained by hydrolyzing an intact antibody molecule [see Morimoto et al., J. Biochem. Biophys. Methods 24:107-117 (1992) and Brennan et al., Science 229:81 (1985)]. In addition, such antigen-binding fragments can also be produced directly from recombinant host cells [reviewed in Hudson, Curr. Opin. Immunol. 11: 548-557 (1999); Little et al., Immunol. Today, 21: 364-370 (2000)]. For example, Fab’ fragments can be obtained directly from host cells, and Fab’ fragments can be chemically conjugated to form F(ab’)2 fragments [Carter et al., Bio / Technology, 10: 163-167 (1992)]. In addition, Fv, Fab or F(ab’)2 fragments can also be isolated directly from the cell culture of recombinant host cells. Those skilled in the art are fully aware of other techniques for preparing such antigen-binding fragments. 【0058】 Isolated nucleic acid molecule In another aspect, the present application also provides the above-mentioned antibody or this antigen-binding fragment or an isolated nucleic acid molecule encoding this heavy chain variable region and / or light chain variable region. 【0059】 In certain embodiments, the isolated nucleic acid molecule comprises a first nucleotide sequence encoding the heavy chain or heavy chain variable region of the antibody or this antigen-binding fragment of the present invention and a second nucleotide sequence encoding the light chain or light chain variable region of the antibody or this antigen-binding fragment, and the first nucleotide sequence and the second nucleotide sequence are present on the same or different isolated nucleic acid molecules. When the first nucleotide sequence and the second nucleotide sequence are present on different isolated nucleic acid molecules, the isolated nucleic acid molecule of the present invention comprises a first nucleic acid molecule comprising the first nucleotide sequence and a second nucleic acid molecule comprising the second nucleotide sequence. 【0060】 Vector In another aspect, the present application provides a vector comprising the above nucleic acid molecule. In certain embodiments, the vector is a cloning vector or an expression vector. 【0061】 In certain embodiments, the vector comprises a first nucleotide sequence encoding the heavy chain or the heavy chain variable region of an antibody or antigen-binding fragment thereof of the present invention and a second nucleotide sequence encoding the light chain or the light chain variable region of the antibody or antigen-binding fragment thereof, and the first nucleotide sequence and the second nucleotide sequence are present on the same or different vectors. When the first nucleotide sequence and the second nucleotide sequence are present on different vectors, the vector of the present invention comprises a first vector comprising the first nucleotide sequence and a second vector comprising the second nucleotide sequence. 【0062】 Host cell In another aspect, the present application also provides a host cell comprising the above nucleic acid molecule or vector. Such host cells include, but are not limited to, prokaryotic cells, such as bacterial cells [e.g., Escherichia coli (E. coli) cells], and eukaryotic cells, such as fungal cells (e.g., yeast cells), insect cells, plant cells, and animal cells (e.g., mammalian cells, such as mouse cells, human cells, etc.). 【0063】 Preparation method In another aspect, the present application provides a method for preparing the above antibody or antigen-binding fragment thereof, comprising culturing the above host cell under conditions that allow expression of the antibody or antigen-binding fragment thereof, and recovering the antibody or antigen-binding fragment thereof from the cell culture of the host cell. 【0064】 Therapeutic use In another aspect, the present application provides a pharmaceutical composition comprising the above antibody or antigen-binding fragment thereof and a pharmaceutically acceptable carrier and / or excipient. 【0065】 In certain exemplary embodiments, pharmaceutically acceptable carriers and / or excipients include sterile injectable solutions (e.g., aqueous or non-aqueous suspensions or solutions). In certain exemplary embodiments, such sterile injectable solutions are selected from the group consisting of water for injection (WFI), bacteriostatic water for injection (BWFI), sodium chloride solutions (e.g., 0.9% (w / v) NaCl), glucose solutions (e.g., 5% glucose), surfactant-containing solutions (e.g., solutions containing 0.01% polysorbate 20), pH buffer solutions (e.g., phosphate buffer solutions), Ringer's solutions, and any combination thereof. 【0066】 In another aspect, the present application provides for the use of the above-described antibody or antigen-binding fragment thereof, isolated nucleic acid molecule, vector, host cell, or pharmaceutical composition in the manufacture of a medicament, which is used for activating CD40, enhancing immune cell activity, enhancing immune response, and / or preventing and / or treating tumors or infections in a subject. 【0067】 In certain embodiments, the immune cells are T cells, B cells, DCs, macrophages, and / or NK cells. 【0068】 In certain embodiments, the immune response is a CD40-mediated immune response. 【0069】 In certain embodiments, the tumor is selected from solid tumors or hematological tumors (e.g., leukemia, lymphoma, myeloma). In certain embodiments, the tumor is selected from solid tumors or lymphomas. 【0070】 In certain embodiments, the tumor is selected from the group consisting of colorectal cancer, colon cancer, bladder cancer, breast cancer, uterine / cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, central nervous system cancer, lymphoma, leukemia, myeloma, sarcoma, and melanoma. 【0071】 In certain embodiments, the infectious disease is selected from the group consisting of viral infections, bacterial infections, fungal infections, and parasitic infections. 【0072】 In certain embodiments, the subject is a mammal, such as a human. 【0073】 In some embodiments, the antibody or antigen-binding fragment thereof is used alone or in combination with a further pharmaceutically active substance. 【0074】 In another aspect, the present application provides a method for enhancing an immune response and / or preventing and / or treating a tumor or an infectious disease in a subject, comprising administering to the subject in need thereof an effective amount of the above antibody or antigen-binding fragment thereof or a pharmaceutical composition. 【0075】 In some embodiments, the immune response is a CD40-mediated immune response. 【0076】 In certain embodiments, the tumor is selected from solid tumors or hematological tumors (e.g., leukemia, lymphoma, myeloma). In certain embodiments, the tumor is selected from solid tumors or lymphomas. 【0077】 In certain embodiments, the tumor is selected from the group consisting of colorectal cancer, colon cancer, bladder cancer, breast cancer, uterine / cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell carcinoma, bone cancer, liver cancer, thyroid cancer, skin cancer, central nervous system tumors, lymphoma, leukemia, myeloma, sarcoma, and melanoma. 【0078】 In certain embodiments, the infectious disease is selected from the group consisting of viral infections, bacterial infections, fungal infections, and parasitic infections. 【0079】 In certain embodiments, the subject is a mammal, such as a human. 【0080】 The antibody or antigen-binding fragment thereof or pharmaceutical composition of the present application can be formulated into any dosage form known in the medical field, for example, tablets, pills, suspensions, emulsions, solutions, gels, capsules, powders, granules, elixirs, troches, suppositories, injections (including injection solutions, sterile powders for injection, and concentrated injection solutions), inhalants, sprays, etc. The preferred dosage form depends on the intended route of administration and therapeutic use. The antibody or antigen-binding fragment thereof or pharmaceutical composition of the present invention must be sterile and stable under the conditions of production and storage. A preferred dosage form is an injection. Such an injection can be a sterile injection solution. For example, a sterile injection solution can be prepared by incorporating the required dose of the antibody or antigen-binding fragment thereof of the present invention and, if appropriate, other desired components (including, but not limited to, pH adjusters, surfactants, adjuvants, ionic strength enhancers, isotonic substances, preservatives, diluents, or any combination thereof) into a suitable solvent and then filter sterilizing. In addition, a sterile injection solution can be prepared as a sterile lyophilized powder (for example, by vacuum drying or freeze drying) to facilitate storage and use. Such a sterile lyophilized powder can be dispersed in a suitable carrier, such as water for injection (WFI), bacteriostatic water for injection (BWFI), sodium chloride solution [for example, 0.9% (w / v) NaCl], glucose solution (for example, 5% glucose), surfactant-containing solution (for example, a solution containing 0.01% polysorbate 20), pH buffer solution (for example, phosphate buffer solution), Ringer's solution, and any combination thereof, before use. 【0081】 The antibody or antigen-binding fragment thereof of the present application or the pharmaceutical composition of the present invention can be administered by any suitable method known in the art, including, but not limited to, oral, buccal, sublingual, intraocular, topical, parenteral, rectal, intrathecal, intracisternal, inguinal, intravesical, external (e.g., powder, ointment or droplet) or nasal routes. However, in many therapeutic uses, the preferred route / method of administration is parenteral administration (e.g., intravenous or bolus injection, subcutaneous injection, intraperitoneal injection, intramuscular injection). Those skilled in the art will understand that the route and / or method of administration will vary depending on the intended purpose. In certain embodiments, the antibody or antigen-binding fragment thereof or the pharmaceutical composition of the present invention is administered by intravenous injection or bolus injection. 【0082】 Detectable uses In another aspect, the present application provides a conjugate comprising the above antibody or antigen-binding fragment thereof and a detectable label that binds to the antibody or antigen-binding fragment thereof. 【0083】 In certain embodiments, the detectable label is selected from the group consisting of enzymes (e.g., horseradish peroxidase or alkaline phosphatase), chemiluminescent reagents (e.g., acridinium ester, luminol and its derivatives or ruthenium derivatives), fluorescent dyes (e.g., fluorescein or fluorescent proteins), radionuclides, and biotin. 【0084】 In another aspect, the present application provides a kit comprising the above antibody or antigen-binding fragment thereof or conjugate. 【0085】 In certain embodiments, the kit comprises the above conjugate. 【0086】 In certain embodiments, the kit includes the above-described antibody or antigen-binding fragment thereof and a secondary antibody that specifically recognizes the antibody or antigen-binding fragment thereof. In certain embodiments, the secondary antibody further includes a detectable label, such as an enzyme (e.g., horseradish peroxidase or alkaline phosphatase), a chemiluminescent reagent (e.g., acridinium ester, luminol and its derivatives or ruthenium derivatives), a fluorescent dye (e.g., fluorescein or fluorescent protein), a radionuclide or biotin. 【0087】 In another aspect, the present application provides a method for detecting the presence or level of CD40 in a sample, including using the above-described antibody or antigen-binding fragment thereof or the above-described conjugate. In certain embodiments, the method is used for therapeutic purposes, diagnostic purposes or non-therapeutic non-diagnostic purposes. 【0088】 In certain embodiments, the method is an immunological assay, such as a Western blot, an enzyme immunoassay (e.g., ELISA), a chemiluminescent immunoassay, a fluorescent immunoassay or a radioimmunoassay. 【0089】 In certain embodiments, the method includes using the above-described conjugate. 【0090】 In certain embodiments, the method includes using the above-described antibody or antigen-binding fragment thereof, and further includes detecting the antibody or antigen-binding fragment thereof using a secondary antibody carrying a detectable label [e.g., an enzyme (e.g., horseradish peroxidase or alkaline phosphatase), a chemiluminescent reagent (e.g., acridinium ester, luminol and its derivatives or ruthenium derivatives), a fluorescent dye (e.g., fluorescein or fluorescent protein), a radionuclide or biotin]. 【0091】 In certain embodiments, the method comprises (1) contacting a sample with an antibody of the invention or an antigen-binding fragment or conjugate thereof, and (2) detecting the formation of an antigen-antibody immune complex or detecting the amount of the immune complex. The formation of the immune complex indicates the presence of CD40 or CD40-expressing cells. 【0092】 In another aspect, the present application also provides the use of the above antibody or an antigen-binding fragment or conjugate thereof in the manufacture of a detection reagent for detecting the presence or level of CD40 in a sample. 【0093】 In certain embodiments, the presence or level of CD40 in a sample is detected with a detection reagent using the above method. 【0094】 In certain embodiments, the sample is a cell sample (e.g., immune cells) derived from a subject (e.g., a mammal, preferably a human or a cynomolgus monkey). 【0095】 Definition of Terms In the present application, unless otherwise stated, scientific and technical terms used herein have meanings commonly understood by those skilled in the art. Moreover, the virological, biochemical, and immunological assays used herein are conventional methods widely used in the corresponding fields. On the other hand, for a better understanding of the present application, definitions and explanations of related terms are provided below. 【0096】 When the terms "for example (for example, such as, e.g.)", "comprising (comprising, including)" or variants thereof are used herein, such terms are not considered to be restrictive, but rather are construed to mean "but without limitation" or "without limitation". 【0097】 Unless the context dictates otherwise, and except as otherwise clearly inconsistent with the context, the terms "a", "an", and "the", as well as similar reference terms in the context in which this application is described (in particular, the context of the following claims), shall be construed to include both the singular and the plural. 【0098】 As used herein, the term "antibody" refers to an immunoglobulin molecule typically composed of two pairs of polypeptide chains, each pair having a light chain (LC) and a heavy chain (HC). Antibody light chains can be classified as κ (kappa) light chains and λ (lambda) light chains. Heavy chains can be classified as μ, δ, γ, α, or ε, and define the antibody isotypes as IgM, IgD, IgG, IgA, and IgE, respectively. In light and heavy chains, the variable and constant regions are connected by a "J" region consisting of approximately 12 or more amino acids, and the heavy chain also contains a "D" region consisting of approximately 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region consists of three domains (CH1, CH2, and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of one domain, CL. This constant domain is not directly involved in the binding of the antibody to the antigen, but mediates various effector functions, such as the binding of the immunoglobulin to host tissues, or various cells of the immune system (e.g., effector cells) and factors including the first component of the classical complement system (Clq). Also, the VH region and the VL region can be subdivided into highly variable regions called complementarity determining regions (CDRs), with more conserved regions called framework regions (FRs) interspersed. V H and V LEach of them consists of three CDRs and four FRs, and is arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions (VH and VL) of each heavy / light chain pair each form an antigen-binding site. The assignment of amino acid regions or domains can follow the definitions described in Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)) or Chothia & Lesk (1987) J. Mol. Biol. 196: 901-917; Chothia et al. (1989) Nature 342: 878-883. 【0099】 As used herein, the terms "complementary determining region" or "CDR" refer to amino acid residues within the antibody variable regions that mediate antigen binding. The variable regions of the heavy and light chains each contain three CDRs, designated CDR1, CDR2, and CDR3. The exact boundaries of such CDRs can be defined according to various numbering schemes known in the art, such as the Kabat numbering scheme (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), the Chothia numbering scheme [Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883] or the IMGT numbering scheme (Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003). For a given antibody, one of ordinary skill in the art can readily identify the CDRs defined by each numbering scheme. Moreover, the correspondence between the various numbering schemes is well known to those of ordinary skill in the art (see, for example, Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003). 【0100】 In the present application, the CDRs contained in the antibody of the present invention or antigen-binding fragment thereof can be determined according to various numbering systems known in the art. In certain embodiments, the CDRs contained in the antibody of the present invention or antigen-binding fragment thereof are determined by the Kabat, Chothia or IMGT numbering system. In certain embodiments, the CDRs contained in the antibody of the present invention or antigen-binding fragment thereof are determined by the numbering method described in the section "Sequence analysis" on page 11 of Lu X, Nobrega RP, Lynaugh H, et al. Deamidation and isomerization liability analysis of 131 clinical-stage antibodies. MAbs. 2019 Jan;11(1):45-57. doi: 10.1080 / 19420862.2018.1548233, which is incorporated herein by reference in its entirety. These CDR L1-L3 refer to positions 24-34, 50-56, and 89-97 respectively, and CDR H1-H3 refer to positions 27-35, 50-65, and 93-102 respectively, and the above positions refer to Kabat positions. 【0101】 As used herein, the term "framework region" or "FR" residues refers to amino acid residues within the antibody variable region other than the CDR residues defined above. 【0102】 The term "antibody" is not limited to any particular method of generating the antibody. This includes, for example, recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. Antibodies can be antibodies of various isotypes, such as IgG (e.g., IgG1, IgG2, IgG3 or IgG4 subtypes), IgA1, IgA2, IgD, IgE or IgM. 【0103】 As used herein, the term “antigen-binding fragment” of an antibody refers to a polypeptide that retains the ability to specifically bind to the same antigen to which the full-length antibody binds and / or competes with the full-length antibody for specific binding to the antigen, and which is also referred to as the “antigen-binding portion”. In general, see Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd ed., Raven Press, N.Y. (1989)), which is hereby incorporated by reference in its entirety for all purposes. Antigen-binding fragments of antibodies can be obtained by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Non-limiting examples of antigen-binding fragments include Fab, Fab’, F(ab’)2, Fd, Fv, complementarity-determining region (CDR) fragments, scFv, diabodies, single-domain antibodies, chimeric antibodies, linear antibodies, nanobodies (technology by Domantis), probodies, and polypeptides containing at least a portion of an antibody sufficient to confer specific antigen-binding ability. A review of engineered variant antibodies is described in Holliger et al., 2005; Nat Biotechnol, 23: 1126-1136. 【0104】 As used herein, the term "full-length antibody" refers to an antibody consisting of two "full-length heavy chains" and two "full-length light chains". In particular, a "full-length heavy chain" refers to a polypeptide chain consisting of, in the direction from the N-terminus to the C-terminus, a heavy chain variable region (VH), a heavy chain constant region CH1 domain, a hinge region (HR), a heavy chain constant region CH2 domain, and a heavy chain constant region CH3 domain, and may also include a heavy chain constant region CH4 domain if the full-length antibody is of the IgE isotype. Preferably, a "full-length heavy chain" is a polypeptide chain consisting of VH, CH1, HR, CH2, and CH3 in the direction from the N-terminus to the C-terminus. A "full-length light chain" is a polypeptide chain consisting of a light chain variable region (VL) and a light chain constant region (CL) in the direction from the N-terminus to the C-terminus. The two pairs of full-length antibody chains are joined together by disulfide bonds between CL and CH1 and between the HRs of the two full-length heavy chains. The full-length antibodies of the present invention may be of a single species, for example, of human origin, and may be chimeric antibodies or humanized antibodies. The full-length antibodies of the present invention include two antigen-binding sites formed by pairs of VH and VL, and such two antigen-binding sites specifically recognize / bind to the same antigen. 【0105】 As used herein, the term "Fd" refers to an antibody fragment consisting of a VH domain and a CH1 domain, the term "dAb fragment" refers to an antibody fragment consisting of a VH domain [Ward et al., Nature 341:544 546 (1989)], the term "Fab fragment" refers to an antibody fragment consisting of a VL domain, a VH domain, a CL domain, and a CH1 domain, the term "F(ab’)2 fragment" refers to an antibody fragment consisting of two Fab fragments connected by a disulfide bridge on the hinge region, and the term "Fab fragment" refers to a fragment obtained by reduction of the disulfide bond connecting the two heavy chain fragments within the F(ab’)2 fragment and consisting of a full light chain and a heavy chain Fd fragment (consisting of a VH domain and a CH1 domain). 【0106】 As used herein, the term "Fv" refers to an antibody fragment consisting of the VL and VH domains of a single arm of an antibody. An Fv fragment is generally considered to be the smallest antibody fragment capable of forming a complete antigen-binding site. Six CDRs are generally thought to confer the antigen-binding specificity of an antibody. However, even the variable region (e.g., an Fd fragment containing only three antigen-specific CDRs), although it may have a lower affinity than the intact binding site, is capable of recognizing and binding to an antigen. 【0107】 As used herein, the term "Fc" refers to an antibody fragment formed by the association of the second and third constant regions of the first heavy chain of an antibody with the second and third constant regions of the second heavy chain of the antibody through disulfide bonds. The Fc fragment of an antibody has a variety of functions but is not involved in antigen binding. 【0108】 As used herein, the term "scFv" refers to a single polypeptide chain containing a VL domain and a VH domain, where the VL and VH are connected via a linker [see, for example, Bird et al., Science. 242:423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Roseburg and Moore, eds., Springer-Verlag, New York, pp. 269-315 (1994)]. Such scFv molecules can have the general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable linkers according to the prior art consist of the GGGGS amino acid repeat sequence or variants thereof. For example, a linker having the amino acid sequence (GGGGS)4 can be used, but variants thereof can also be used [Holliger et al. (1993), Proc. Natl. Acad. Sci. USA 90: 6444-6448]. Other linkers useful in the present application are described in Alfthan et al. (1995), Protein Eng. 8:725-731, Choi et al. (2001), Eur. J. Immunol. 31:94-106, Hu et al. (1996), Cancer Res. 56:3055-3061, Kipriyanov et al. (1999), J. Mol. Biol. 293:41-56, and Roovers et al. (2001), Cancer Immunol. In some cases, a disulfide bond may also be present between the VH and VL of the scFv. In certain embodiments of the present application, the scFv can form a di-scFv, which refers to an antibody formed by connecting two or more individual scFvs in series.In certain embodiments of the present application, the scFv is capable of forming (scFv)2, which refers to an antibody formed by connecting two or more individual scFvs in parallel. 【0109】 As used herein, the term "diabody" refers to an antibody in which the VH domain and the VL domain are expressed on a single polypeptide chain, but the linker used is too short to allow pairing between the two domains on the same chain, which forces the domains to pair with complementary domains of other chains to generate two antigen-binding sites [see, for example, Holliger P. et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993), and Poljak R. J. et al., Structure 2:1121-1123 (1994)]. 【0110】 As used herein, the term "single-domain antibody (sdAb)" has the same meaning as commonly understood by those skilled in the art and refers to an antibody fragment consisting of a single monomeric variable antibody domain (e.g., a single heavy-chain variable region) that retains the ability to specifically bind to the same antigen to which a full-length antibody binds. Single-domain antibodies are also referred to as nanobodies. 【0111】 Each of the above antibody fragments retains the ability to specifically bind to the same antigen to which a full-length antibody binds and / or competes with a full-length antibody for specific binding to the antigen. 【0112】 Antigen-binding fragments of an antibody (e.g., the above antibody fragments) can be obtained from a given antibody (e.g., an antibody provided by the present application) using conventional techniques known to those skilled in the art (e.g., recombinant DNA technology or enzymatic or chemical fragmentation methods), and the specificity of the antigen-binding fragment of the antibody is screened in the same manner as for the intact antibody. 【0113】 As used herein, the term "chimeric antibody" refers to an antibody in which a portion of the light and / or heavy chain is derived from an antibody (which may be from a particular species or belong to a particular antibody class or subclass), and the other portion of the light or / and heavy chain is derived from another antibody (which may be from the same or a different species or belong to the same or a different antibody class or subclass), but nevertheless retains binding activity to the target antigen [U.S. Patent No. 4,816,567 by Cabilly et al.; Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851 - 6855 (1984)]. In certain embodiments, the term "chimeric antibody" may include an antibody in which the heavy chain variable region and the light chain variable region of the antibody are derived from a first antibody, and the heavy chain constant region and the light chain constant region of the antibody are derived from a second antibody. 【0114】 As used herein, the term "identity" is used to refer to sequence identity between two polypeptides or between two nucleic acids. To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps are introduced into the first amino acid sequence or nucleic acid sequence to obtain the best alignment with the second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at the corresponding amino acid positions or nucleotide positions are then compared. The molecule is identical at a position if the position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence. The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., percent identity = number of identical overlapping positions / total number of positions × 100%). In certain embodiments, both sequences are of the same length. 【0115】 In addition, the determination of percent identity between two sequences can be accomplished using mathematical algorithms. A non-limiting example of a mathematical algorithm for comparing two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. U.S.A. 87:2264-2268, which was modified by Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. U.S.A. 90:5873-5877. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403. 【0116】 As used herein, the term "variant" also refers to a polypeptide or peptide that includes an amino acid sequence that has been altered by introducing an amino acid residue substitution, deletion, or addition in the context of a polypeptide (including a polypeptide). In some cases, the term "variant" also refers to a polypeptide or peptide that has been modified (i.e., by covalently attaching any type of molecule to the polypeptide or peptide). For example, without limitation, a polypeptide can be modified by, for example, glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, conjugation to a cell ligand or another protein, etc. Derivatized polypeptides or peptides can be produced using techniques known to those of skill in the art by chemical modification, including, without limitation, specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicaamycin, etc. Moreover, the variant has a similar, identical, or improved function relative to the polypeptide or peptide from which it is derived. 【0117】 As used herein, the term "specific binding" refers to a non-random binding reaction between two molecules, for example, the reaction between an antibody and an antigen to be targeted. The strength or affinity of a specific binding interaction can be represented by the equilibrium dissociation constant (K D ) of the interaction. In this application, "KD The term " " refers to the dissociation equilibrium constant of the antibody-antigen specific interaction and is used to describe the binding affinity between an antibody and an antigen. The smaller the equilibrium dissociation constant, the stronger the antibody-antigen binding and the higher the affinity between the antibody and the antigen. 【0118】 The specific binding characteristics between two molecules can be determined using methods known in the art. One approach involves measuring the rates at which the antigen-binding site / antigen complex forms and dissociates. The "association rate constant" (k a or k on ) and the "dissociation rate constant" (k dis or k off ) can both be calculated from the concentrations and actual rates of association and dissociation (see Malmqvist M, Nature, 1993, 361 :186-187). The ratio of k dis / k on is equal to the dissociation constant K D (see Davies et al., Annual Rev Biochem, 1990; 59:439-473). The K D value, the k on value, and the k dis value can be measured by any effective method. In certain embodiments, the dissociation constant can be measured by surface plasmon resonance (SPR) using equipment from Biacore. Additionally, bioluminescence interference or Kinexa can be used to measure the dissociation constant. 【0119】 As used herein, the detectable label of the present application can be any substance detectable by fluorescent, spectroscopic, photochemical, biochemical, immunological, electrical, optical, or chemical means. Such labels are well known in the art and include, by way of example, enzymes (e.g., horseradish peroxidase, alkaline phosphatase, beta-galactosidase, urease, glucose oxidase, etc.), radionuclides (e.g., 3 H, 125 I, 35 S, 14 C or 32P), fluorescent dyes [e.g., fluorescein isothiocyanate (FITC), fluorescein, tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin (PE), Texas Red, rhodamine, quantum dots or cyanine dye derivatives (e.g., Cy7, Alexa750)], luminescent substances (e.g., chemiluminescent substances, e.g., acridinium ester, luminol and its derivative compounds, ruthenium derivatives, e.g., ruthenium terpyridyl), magnetic beads [e.g., Dynabeads (registered trademark)], thermometric markers, e.g., colloidal gold or colored glass, or plastic (e.g., polystyrene, polypropylene, latex, etc.) beads, and biotin for binding to avidin (e.g., streptavidin) modified with the above labels are included, but not limited to these. 【0120】 As used herein, the term "vector" refers to a nucleic acid delivery medium into which a polynucleotide can be inserted. When a vector is capable of expressing a protein encoded by the inserted polynucleotide, the vector is called an expression vector. A vector can be introduced into a host cell by transformation, transduction or transfection such that the genetic material elements it carries are capable of being expressed within the host cell. Vectors are well known to those skilled in the art and include, but are not limited to, plasmids; phagemids; cosmids; artificial chromosomes, e.g., yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs) or Pl-derived artificial chromosomes (PACs); phages, e.g., λ phage or M13 phage, and animal viruses, etc. Animal viruses that can be used as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpesviruses (e.g., herpes simplex virus), poxviruses, baculoviruses, papillomaviruses, papovaviruses (e.g., SV40). A vector can include various expression regulatory elements, including, but not limited to, promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. In addition, a vector can also include an origin of replication. 【0121】 As used herein, the term "host cell" refers to a cell that can be used for the introduction of a vector, and includes, but is not limited to, prokaryotic cells such as Escherichia coli or Bacillus subtilis, fungal cells such as yeast cells or Aspergillus, insect cells such as S2 Drosophila cells or Sf9, or animal cells such as fibroblasts, CHO cells, COS cells, NSO cells, HeLa cells, BHK cells, HEK293 cells or human cells. 【0122】 As used herein, the term "conservative substitution" refers to an amino acid substitution that does not adversely affect or change the predicted properties of a protein / polypeptide containing an amino acid sequence. For example, conservative substitutions can be introduced by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative substitutions of amino acids include substituting an amino acid residue with another amino acid residue having a similar side chain, e.g., a side chain that is physically or functionally similar to the corresponding amino acid residue (e.g., having chemical properties such as similar size, shape, charge, ability to form covalent or hydrogen bonds, etc.). Families of amino acid residues having similar side chains are defined in the art. Such families include amino acids having basic side chains (e.g., lysine, arginine, and histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, it is preferred to substitute the corresponding amino acid residue with another amino acid residue from the same side chain family. Methods for identifying conservative substitutions of amino acids are well known in the art [see, e.g., Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10):879-884 (1999); and Burks et al. Proc. Natl Acad. Set USA 94:412-417 (1997), which are incorporated herein by reference]. 【0123】 The 20 standard amino acids included in this specification are described according to conventional usage. See, for example, Immunology-A Synthesis (2nd Edition, E. S. Golub and D. R. Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. In this application, the terms "polypeptide" and "protein" have the same meaning and are used interchangeably. Further, in this application, amino acids are generally represented by the one-letter and three-letter abbreviations well known in the art. For example, alanine can be represented by A or Ala. 【0124】 As used herein, the term "pharmaceutically acceptable carrier and / or excipient" refers to carriers and / or excipients that are pharmacologically and / or physiologically compatible with the subject and the active ingredient, which are well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and include, but are not limited to, pH adjusting substances, surfactants, adjuvants, ionic strength enhancers, diluents, substances for maintaining osmotic pressure, substances for delaying absorption, and preservatives. For example, pH adjusting substances include, but are not limited to, phosphate buffers. Surfactants include cationic, anionic or nonionic surfactants, such as, but not limited to, Tween-80. Ionic strength enhancers include, but are not limited to, sodium chloride. Preservatives include various antibacterial and antifungal substances, such as, but not limited to, parabens, chlorobutanol, phenol, sorbic acid, etc. Substances for maintaining osmotic pressure include, but are not limited to, sugars, NaCl, etc. Substances for delaying absorption include, but are not limited to, monostearic acid and gelatin. Diluents include, but are not limited to, water, aqueous buffers (e.g., buffered saline), alcohols, and polyols (e.g., glycerol). Preservatives include various antibacterial and antifungal substances, such as, but not limited to, thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid, etc.A stabilizer has the meaning generally understood by those skilled in the art to be capable of stabilizing the desired activity of the active ingredient in a medicament, and includes, but is not limited to, sodium glutamate, gelatin, SPGA, saccharides (e.g., sorbitol, mannitol, starch, sucrose, lactose, dextran or glucose), amino acids (e.g., glutamic acid, glycine), proteins (e.g., dried whey, albumin or casein) or their degradation products (e.g., lactalbumin hydrolysate), etc. In certain exemplary embodiments, a pharmaceutically acceptable carrier or excipient includes a sterile injection solution (e.g., an aqueous or non-aqueous suspension or solution). In certain exemplary embodiments, such sterile injection solutions are selected from the group consisting of water for injection (WFI), bacteriostatic water for injection (BWFI), sodium chloride solutions [e.g., 0.9% (w / v) NaCl], glucose solutions (e.g., 5% glucose), surfactant-containing solutions (e.g., solutions containing 0.01% polysorbate 20), pH buffer solutions (e.g., phosphate buffer solutions), Ringer's solutions, and any combination thereof. 【0125】 As used herein, the term "prevent" refers to a method of preventing or delaying the onset of a disease or symptom or syndrome in a subject. As used herein, the term "treat" refers to a method of achieving a beneficial or desired clinical outcome. For the purposes of this application, beneficial or desired clinical outcomes include, but are not limited to, symptom relief, reduction in the degree of disease, stabilization of the disease state (i.e., non-worsening), delay or slowing of disease progression, recovery or alleviation of the disease state, and alleviation (partial or complete, detectable or undetectable) of symptoms. In addition, "treat" can also refer to an extension of survival compared to survival expected in the absence of treatment. 【0126】 As used herein, the term "subject" refers to a mammal, e.g., a human. In certain embodiments, the subject (e.g., a human) has or is at risk of having a tumor, an infection or an autoimmune disease. 【0127】 As used herein, the term "effective amount" refers to an amount sufficient to achieve or at least partially achieve the desired effect. For example, an effective amount for preventing a disease (e.g., a tumor, an infectious disease or an autoimmune disease) refers to an amount sufficient to prevent, inhibit or delay the onset of the disease, and an effective amount for treating a disease refers to an amount sufficient to cure or at least partially prevent the disease and its complications in a patient already having the disease. The determination of such an effective amount is well within the ability of those skilled in the art. For example, the amount effective for therapeutic use depends on the severity of the disease to be treated, the overall state of the patient's own immune system, the general condition of the patient, such as age, weight, and gender, the method of administering the drug, and other treatments administered simultaneously, etc. 【Advantages of the Invention】 【0128】 The present application provides a fully human antibody against CD40 that has a high binding affinity for CD40, shows cross-reactivity with human CD40 and cynomolgus monkey CD40, and is capable of either blocking or not blocking the binding to the ligand CD40L of CD40. 【Brief Description of the Drawings】 【0129】 【Figure 1】 Figure 1 shows the binding activity of a parental anti-CD40 candidate molecule to CHO cells expressing human CD40. 【Figure 2】 Figure 2 shows the activity of a parental anti-CD40 candidate molecule in the activation of a reporter gene in CD40 signal transduction in the absence of bead crosslinking. 【Figure 3】 Figure 3 shows the activity of a parental anti-CD40 candidate molecule in the activation of a reporter gene in CD40 signal transduction in the presence of bead crosslinking. 【Figure 4】 Figure 4 shows the binding activity of a progeny anti-CD40 candidate molecule to CHO cells expressing human CD40. 【Figure 5】 Figure 5 shows the binding activity of a progeny anti-CD40 candidate molecule to CHO cells expressing cynomolgus monkey CD40. 【Figure 6】Figure 6 shows the activity of a progeny anti-CD40 candidate molecule in the activation of a reporter gene for CD40 signal transduction in the presence of an anti-Fc crosslink. 【Figure 7】 Figure 7 shows the activity of a progeny anti-CD40 candidate molecule in the activation of a reporter gene for CD40 signal transduction in the absence of an anti-Fc crosslink. **DETAILED DESCRIPTION OF THE INVENTION** 【0130】 Embodiments of the present invention will be described in detail below with reference to the accompanying drawings and examples. Those skilled in the art will understand that the subsequent drawings and examples are merely used to illustrate the present invention and do not limit the scope of the present invention. Various objects and advantageous aspects of the present invention will become apparent to those skilled in the art from the accompanying drawings and the detailed description of the following preferred embodiments. 【0131】 Sequence Information The description of the sequences included by this application is presented in the table below. 【0132】 **Table 1-1** **Table 1-2** 【0133】 Here, the present invention will be described with reference to the following examples, which are intended to illustrate the present invention (but not limit the present invention). 【0134】 Unless otherwise specified, the molecular biology experimental methods and immunoassay methods used in this application are basically carried out with reference to the methods described by J. Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989 and F. M. Ausubel et al., Compiled Laboratory Guide to Molecular Biology, 3rd Edition, John Wiley & Sons, Inc., 1995. The use of restriction enzymes followed the conditions recommended by the manufacturer. Those skilled in the art will understand that in the examples, the present invention is described by way of example and is not intended to limit the scope of the claimed invention. 【Example】 【0135】 [Example 1] Preparation of Antibody 【0136】 1.1 Yeast Display Technology for Screening of Anti-CD40 Fully Human Antibodies Based on the yeast display antibody library (Adimab), eight natural synthetic libraries were amplified according to the existing methods (International Publication No. WO2009036379, International Publication No. WO2010105256, International Publication No. WO2012009568), and the diversity of each library reached 1×10 9 For simplicity, the first two rounds of screening were carried out using the Miltenyi MACS system for magnetic bead cell sorting. First, yeast cells derived from the library (about 1×10 per library) in FACS wash buffer (phosphate buffered saline containing 0.1% BSA) 10The cells were incubated for 15 minutes at room temperature with a buffer containing 10 nM biotinylated human CD40-Fc fusion protein (purchased from ACRO, catalog number: CD0-H5253, and the amino acid sequence of this human CD40 is shown as SEQ ID NO: 71). The cells were washed once with 50 mL of pre-cooled FACS wash buffer, then resuspended in 40 mL of buffer, and after adding 500 μL of streptavidin magnetic beads (Miltenyi LS), incubated for 15 minutes at room temperature. After centrifuging at 1000 rpm for 5 minutes to discard the supernatant, the cells were resuspended in 5 mL of FACS wash buffer, and the cell supernatant was added to the Miltenyi LS column. After addition, the column was washed three times with 3 mL of FACS wash buffer each time. The Miltenyi LS column was removed from the magnetic field, elution was carried out with 5 mL of growth medium, and the eluted yeast cells were collected and grown overnight at 37°C. 【0137】 In the next round of sorting using flow cytometry, then, approximately 1×10 yeast cells obtained by screening with the MACS system 8 The cells were washed three times with FACS buffer and cultured at room temperature in a solution containing 100 nM biotinylated CD40 monomer protein (purchased from ACRO, catalog number: CD0-H5228, and the amino acid sequence of this CD40 is shown as SEQ ID NO: 71) or 10 nM CD40-Fc fusion protein. After discarding the culture medium, the cells were washed twice with FACS wash buffer, then mixed with LC-FITC (FITC-labeled anti-human immunoglobulin kappa light chain antibody, purchased from SouthernBiotech) (1:100 dilution) and mixed with SA-633 (streptavidin-633, Molecular Probes) (1:500 dilution) or SA-PE (streptavidin-PE, Sigma) (1:50 dilution), and incubated at 4°C for 15 minutes. The cells were washed twice with pre-cooled FACS wash buffer, resuspended in 0.4 mL of buffer, and then transferred to a separation tube with a filter. The cells were stored using a FACS ARIA (BD Biosciences). 【0138】 Yeast cells expressing the anti-CD40 antibody obtained through screening were induced, and the anti-CD40 antibody (full-length antibody) was secreted by shaking at 30°C for 48 hours. After completion of the induction, the yeast cells were removed by centrifugation at 1300 rpm for 10 minutes, and the supernatant was collected. The anti-CD40 antibody in the supernatant was purified using Protein A, eluted with an acetic acid solution at pH 2.0, and the anti-CD40 antibody was recovered. Through this screening, anti-CD40 antibodies ADI-47754, ADI-47720, and ADI-47741 were obtained. 【0139】 1.2 Affinity optimization of anti-human CD40 antibody To obtain further high-affinity antibodies, antibodies ADI-47754 and ADI-47720 were optimized by the following method without limitation. 【0140】 1.3 CDRH1 / CDRH2 screening The CDRH3 gene, which is the parental molecule, was constructed with a diversity of 1×10 8 within the CDRH1 / CDRH2 gene library and subjected to 4 rounds of screening. The first 2 rounds were used for the MACS method, and the 3rd and 4th rounds were used for the FACS method to apply affinity pressure to the antibody-antigen complex and screen for high-affinity antibodies. 【0141】 Yeast cells expressing the anti-CD40 antibody obtained through screening were cultured with shaking at 30°C for 48 hours to obtain the CD40 antibody. After completion of the expression induction, the yeast cells were removed by centrifugation at 1300 rpm for 10 minutes, and the supernatant was collected. The anti-CD40 antibody in the supernatant was purified using Protein A, eluted with an acetic acid solution at pH 2.0, and the anti-CD40 antibody was recovered. The corresponding Fab fragment was obtained by using papain and purified using KappaSelect (GE Healthcare). 【0142】 1.4 VHmut screening In this method, mutations were introduced into the heavy chain region by conventional error-prone PCR. During the PCR process, the probability of base mismatches was increased to approximately 0.01 bp by using 1 μM of the highly mutagenic base analogs dPTP and 8-oxo-dGTP. 【0143】 The products obtained by error-prone PCR were constructed by homologous recombination into a vector containing the heavy chain constant region. Using this method, a secondary library with a capacity of 1×10 7 was obtained under selection pressure including CD40 antigen titer, unlabeled antigen competition, and competition using the parental antibody. The library was then subjected to three rounds of screening using the FACS method to obtain yeast cells expressing the corresponding antibodies. 【0144】 Table 2 shows the relationship between each antibody progeny after affinity maturation and the parental antibody. 【0145】 【Table 2】 【0146】 1.5 Expression and purification of antibodies The sequences and names of various anti-human CD40 antibody clones involved in this application are shown in Table 3. This includes the parental antibody ADI-47741 not mentioned in Table 2. 【0147】 The antibodies used in the examples were expressed and purified by HEK293 cells. Certain operations are as follows: The pcDNA3.1 vector carrying the coding sequences of the heavy and light chains of the antibody was transfected into HEK293 cells using the chemical transfection method (the amino acid sequence of the heavy chain constant region encoded by the vector is shown as SEQ ID NO: 72, and the amino acid sequence of the light chain constant region is shown as SEQ ID NO: 73), and cultured for 7 days under the conditions of 37 °C and 8% CO2. The cell culture was collected and centrifuged at 13,000 rpm for 20 minutes. The supernatant was taken out, and the antibody was purified by Protein A. The purity of the antibody was detected by SEC, and the endotoxin content was adjusted simultaneously. 【0148】 [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] 【0149】 [Example 2] Detection of Antibody Affinity 【0150】 The binding and dissociation constants (K D ) of the antibodies involved in the present application with respect to the binding to human CD40 and cynomolgus monkey CD40, and whether they block the binding to the ligand of human CD40 (human CD40L) were determined using optical biosensor interference technology (ForteBio). The amino acid sequence of human CD40 is shown as SEQ ID NO: 71, the amino acid sequence of cynomolgus monkey CD40 is shown as SEQ ID NO: 74, and the amino acid sequence of human CD40L is shown as SEQ ID NO: 75. The Fortebio affinity assay was performed according to the existing method [Este, P et al. High throughput solution-based measurement of antibody-antigen affinity and epitope binning. Mabs, 2013.5(2):p.270-8]. 【0151】 Measurement of the monovalent affinity of candidate antibody Fab fragments for human and cynomolgus macaque CD40-Fc proteins: After offline equilibration of the sensor in the assay buffer for 20 minutes, online detection was performed for 120 seconds to establish a baseline. Subsequently, the CD40-Fc antigen was added to the AHQ sensor to a thickness of 1 nm for affinity detection. The sensor with the added antigen was incubated with a 100 nM Fab solution until a plateau was reached, and then the sensor was transferred to the assay buffer for at least 2 minutes to measure the dissociation rate. Kinetic analysis was performed using a 1:1 binding model. 【0152】 Measurement of the bivalent affinity of intact antibodies for human and cynomolgus macaque CD40-Fc proteins: The sensor was equilibrated offline in the assay buffer for 20 minutes, and then online detection was used for 120 seconds to establish a baseline. An intact CD40 antibody was added to the AHQ sensor to a thickness of 1 nm for affinity detection. The sensor with the added antibody was further incubated for 10 minutes with a high concentration of irrelevant intact antibody solution to saturate and block the Fc binding sites on the AHQ sensor. Subsequently, the blocked sensor was incubated with a 100 nM CD40-Fc antigen until a plateau was reached, and then the sensor was transferred to the assay buffer for at least 2 minutes to measure the dissociation rate. Kinetic analysis was performed using a 1:1 binding model. 【0153】 In the above-described measurement method, the K D values of each parental antibody molecule and this mutant molecule are shown in Table 4. 【0154】 From the results shown in the charts and tables, the following can be understood: (1) After affinity maturation, the monovalent affinity of each progeny molecule for the human CD40 protein was significantly improved compared to the corresponding parental molecule. (2) After affinity maturation, the monovalent affinity of each progeny molecule for the cynomolgus macaque CD40 protein was significantly improved compared to the corresponding parental molecule. 【0155】 [Table 4] 【0156】 [Example 3] Binding of the parental anti-CD40 antibody to human CD40-highly expressing CHO cells 【0157】 The binding activity of the parental anti-CD40 antibody to human CD40-expressing CHO cells was detected by flow cytometry. In this assay, the light chain amino acid sequence of the control antibody CP870893-IgG2 is shown as SEQ ID NO: 80, and the heavy chain amino acid sequence is shown as SEQ ID NO: 81. 【0158】 Specifically, after transfecting the human CD40 cDNA cloned into the MCS of the pCHO1.0 vector (Invitrogen) and applying a selection pressure, CHO-S cells highly expressing human CD40 (CHO-huCD40 cells) were generated. The proliferated human CD40-highly expressing cells were adjusted to an appropriate cell density and seeded on a 96-well flow plate. After centrifugation, the cells were incubated with serially diluted samples at 4°C for 30 minutes. Then, the cells were washed twice with PBS and incubated with a fluorescent secondary antibody diluted to an appropriate concentration at 4°C for 30 minutes. After two washes with PBS, the cells were resuspended in PBS, and the fluorescence signal was detected on a CytoFlex flow cytometer to calculate the corresponding MFI. The results are shown in Figure 1. 【0159】 The following conclusions can be drawn from the experimental results: All of the parental anti-CD40 antibodies ADI-47754, ADI-47720, and ADI-47741 show good monovalent binding activity to CHO cells highly expressing human CD40. 【0160】 [Example 4] Activation of the human CD40 signaling pathway by the parental anti-CD40 antibody 【0161】 The activity of the parental anti-CD40 antibody in the activation of the CD40 signaling pathway was detected using an NFκB-Luc2P reporter gene assay with or without cross-linking by protein G magnetic beads. 【0162】 Specifically, after transfecting U2OS cells that highly express human CD40 with the NFκB-Luc2P vector (Promega, catalog number: 11501ES03), a selection pressure was applied to generate NFκB-Luc2P-U2OS cells. Parental antibodies at final concentrations of 10 μg / ml, 1 μg / ml, 0.1 μg / ml, 0.01 μg / ml, and 0.001 μg / ml were added to NFκB-Luc2P-U2OS cells together with 0.25 μL / well of Pierce™ Protein G Magnetic Beads (Thermo Scientific, catalog number: 88847). The cells were then incubated in a cell culture incubator for 4 hours to examine the activation of CD40 downstream signaling induced by the antibody in the presence of bead cross-linking. In addition, antibodies at corresponding concentrations were added to NFκB-Luc2P-U2OS cells and incubated in a cell culture incubator for 4 hours to investigate the activation of CD40 downstream signaling induced by the antibody alone in the absence of bead cross-linking. 【0163】 The following conclusions can be drawn from the experimental results: The parental clones ADI-47754, ADI-47720, and ADI-47741 are unable to activate the CD40 downstream signaling pathway without Fc cross-linking, and with bead cross-linking, all clones are capable of activating the CD40 downstream signaling pathway, and the signaling activity is enhanced in all antibodies with bead cross-linking. The positive control antibody CP-870893-G2-WT is capable of activating the CD40 downstream signaling pathway independently of anti-Fc cross-linking. The detailed results can be found in Tables 5-7 and Figures 2 and 3. 【0164】 【Table 5】 【0165】 【Table 6】 【0166】 【Table 7】 【0167】 [Example 5] Binding of a progeny anti-CD40 antibody to human / cynomolgus CD40-expressing CHO cells 【0168】 Based on the results of Example 4, the affinities of ADI-47720, ADI-47741, and ADI-47754 were optimized. These are dependent on bead crosslinking to activate CD40 signaling when visualized by a reporter gene, yielding the antibodies shown in Table 2. 【0169】 ADI-47720, ADI-47741, ADI-47754, and their progeny antibodies ADI-55136, ADI-55140, ADI-55142, ADI-55144-2, ADI-55145, ADI-55147, ADI-55164, ADI-55168 were selected for investigation by flow cytometry of their binding activity to human / cynomolgus CD40-high expressing cells. 【0170】 Specifically, after transfecting the pCHO1.0 vector (Invitrogen) with human / cynomolgus CD40 cDNA cloned into the MCS and applying a selection pressure, CHO-S cells highly expressing human / cynomolgus CD40 were generated. The expanded CD40-high expressing cells were adjusted to an appropriate cell density and seeded on a 96-well flow plate. After centrifugation, the cells were incubated with serially diluted samples at 4°C for 30 minutes, then the cells were washed twice with PBS and then incubated with a fluorescent secondary antibody diluted to an appropriate concentration at 4°C for 30 minutes. After two washes with PBS, the cells were resuspended in PBS and the fluorescent signal was detected on a CytoFlex flow cytometer to calculate the corresponding MFI. 【0171】 The following conclusions can be drawn from the experimental results: Compared with the parental antibody, the binding activity of the progeny anti-CD40 antibody to human / cynomolgus CD40-high expressing cells was improved to some extent, as shown in Tables 8-9 and Figures 4-5. 【0172】 [Table 8] 【0173】 [Table 9] 【0174】 [Example 6] Activation of the human CD40 signaling pathway by a progeny anti-CD40 antibody 【0175】 The activities of the parental anti-CD40 antibody and the progeny anti-CD40 antibody in the activation of CD40 signaling were detected using an NFκB-Luc2P reporter gene assay with or without crosslinking by the F(ab)2 fragment of goat anti-human IgG-Fc (Jackson, catalog number: 109-036-098). 【0176】 Specifically, after transfecting Jurkat cells highly expressing human CD40 with the NFκB-Luc2P vector (Promega, catalog number: 11501ES03), a selection pressure was applied to generate CD40-NFκB-Jurkat cells. The parental antibody and the progeny antibody at a final concentration of 200 nM with a 3-fold dilution over 11 concentrations were added to CD40-NFκB-Jurkat cells at a molar ratio of 1:2 together with the F(ab)2 fragment of goat anti-human IgG-Fc (Thermo Scientific, catalog number: 88847), and incubated in a cell culture incubator for 6 hours to investigate the activation of CD40 downstream signaling induced by the antibody in the presence of anti-Fc fragment crosslinking. In addition, the corresponding concentrations of the antibody were added to CD40-NFκB-Jurkat cells and incubated in a cell culture incubator for 6 hours to examine the activation of the CD40 downstream signaling pathway induced by the antibody alone in the absence of anti-Fc fragment crosslinking. 【0177】 The experimental results are shown in FIGS. 6 and 7. From these results, it can be understood that the parental antibodies ADI-47720, ADI-47741, ADI-47754, and the progeny antibodies ADI-55142, ADI-55144-2, ADI-55147, ADI-55164, ADI-55168 are unable to activate the CD40 downstream signaling pathway without anti-Fc crosslinking, but can activate the CD40 downstream signaling pathway to varying degrees with anti-Fc crosslinking. The positive control antibody CP-870893-G2-WT can activate the CD40 downstream signaling pathway independently of anti-Fc crosslinking. In addition, the control antibody CP-870893-G2-WT has been reported to cause severe hepatotoxicity in a clinical setting as a crosslinking-independent CD40 agonist, and this clinical trial was terminated. 【0178】 Although specific embodiments of the present application have been described in detail, those skilled in the art can make various modifications and changes in detail based on all the teachings disclosed, and understand that such changes are all within the protection scope of the present application. The entire present application is provided by the appended claims and their equivalents.

Claims

[Claim 1] An antibody or antigen-binding fragment capable of specifically binding to CD40, (1) The following three complementarity-determining regions (CDRs) of the heavy chain variable region (VH): (a) FTFX １ X ２ YX ３ MH (SEQ ID NO: 76), GSISSYYWS (SEQ ID NO: 6) VH CDR1 having a structure selected from the group consisting of, (b) VIX ４ YX ５ X ６ X ７ X ８ KYYAX ９ SVKG (SEQ ID NO: 77), GISWSSX １０ SIX １１ YADSVKG (SEQ ID NO: 78), SIYYSSGSTNYNPSLKS (SEQ ID NO: 22) VH CDR2 having a structure selected from the group consisting of, (c) ARDTSRGHDI (SEQ ID NO: 23), AKDPTATTTGARGYFDL (SEQ ID NO: 24), ARDANYYKWHPY (SEQ ID NO: 25) VH CDR3 having a structure selected from the group consisting of the above and / or (2) The following three complementarity-determining regions (CDRs) of the light chain variable region (VL): (d) X １２ ASQSVSX １３ X １４ YLA (Sequence ID 79), X １５ ASX １６ X １７ ISSWLA (Sequence ID 7) VL CDR1 having a structure selected from the group consisting of, (e) GASX １８ X １９ X ２０ X ２１ (Sequence No. 8), AASX ２２ LX ２３ S (Sequence No. 9) VL CDR2 having a structure selected from the group consisting of, (f) X ２４ X ２５ YX ２６ DYPPFT (Sequence ID 10), QQX ２７ X ２８ SX ２９ PPT (Sequence No. 11) VL CDR3 having a structure selected from the group consisting of the above Includes, X １ However, (i) amino acid residues S, E, G, D and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２ However, (i) amino acid residues S, K, D, T and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ３ However, (i) amino acid residues G, A and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ４ However, (i) amino acid residues S and H and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ５ However, (i) amino acid residues E, H and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ６ However, (i) amino acid residues G, A and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ７ However, (i) amino acid residues S, E, T, V and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ８ However, (i) amino acid residues N, S, I, K and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ９ However, (i) amino acid residues D and E and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １０ However, (i) amino acid residues G and D and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １１ However, (i) amino acid residues G and H and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １２ However, (i) amino acid residues R and Q and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １３ However, (i) amino acid residues Y and S and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １４ However, (i) amino acid residues S, D, N and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １５ However, (i) amino acid residues R and E and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １６ However, (i) amino acid residues Q, K and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １７ However, (i) amino acid residues G and V and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １８ However, (i) amino acid residues S, T, A and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １９ However, (i) amino acid residues R, L and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２０ However, (i) amino acid residues N, A and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２１ However, (i) amino acid residues T and N and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２２ However, (i) amino acid residues S and Y and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２３ However, (i) amino acid residues E and Q and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２４ However, (i) amino acid residues S, G, Q and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２５ However, (i) amino acid residues E, Q, D and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２６ However, (i) amino acid residues A and S and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２７ However, (i) amino acid residues R, H, V and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２８ However, (i) amino acid residues S, L, A and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２９ However, (i) amino acid residues F, Y and (ii) amino acid residues selected from the group consisting of conserved substitutions for (i), An antibody or this antigen-binding fragment. [Claim 2] (1) The following three complementarity-determining regions (CDRs) of the heavy chain variable region (VH): (a) VH CDR1 having the structure shown in FTFX 1 X 2 YX 3 MH (Sequence ID 76), (b) VH CDR2 having the structure shown in GISWSSX 10 SIX 11 YADSVKG (Sequence ID 78), (c) VH CDR3 having the structure shown in AKDPTATTTGARGYFDL (Sequence ID 24) and / or (2) The following three complementarity-determining regions (CDRs) of the light chain variable region (VL): (d) VL CDR1 having the structure shown in X 15 ASX 16 X 17 ISSWLA (Sequence ID 7), (e) VL CDR2 having the structure shown in AASX 22 LX 23 S (Sequence ID 9), (f) VL CDR3 having the structure shown in QQX 27 X 28 SX 29 PPT (Sequence No. 11) Includes, X1 is selected from the group consisting of (i) amino acid residue D and (ii) amino acid residues that are conservative substitutions for (i), X2 is selected from the group consisting of (i) amino acid residues D and T and (ii) amino acid residues that are conservative substitutions for (i), X3 is selected from the group consisting of (i) amino acid residue A and (ii) amino acid residues that are conservative substitutions for (i), X 10 is selected from the group consisting of (i) amino acid residues G and D and (ii) amino acid residues that are conservative substitutions for (i), X 11 is selected from the group consisting of (i) amino acid residues G and H and (ii) amino acid residues that are conservative substitutions for (i), X 15 is selected from the group consisting of (i) amino acid residues R and E and (ii) amino acid residues that are conservative substitutions for (i), X 16 is selected from the group consisting of (i) amino acid residues Q and K and (ii) amino acid residues that are conservative substitutions for (i), X 17 is selected from the group consisting of (i) amino acid residues G and V and (ii) amino acid residues that are conservative substitutions for (i), X 22 is selected from the group consisting of (i) amino acid residues S and Y and (ii) amino acid residues that are conservative substitutions for (i), X 23 is selected from the group consisting of (i) amino acid residues E and Q and (ii) amino acid residues that are conservative substitutions for (i), X 27 is selected from the group consisting of (i) amino acid residues R and H and (ii) amino acid residues that are conservative substitutions for (i), X 28 is selected from the group consisting of (i) amino acid residues S and A and (ii) amino acid residues that are conservative substitutions for (i), X 29 is selected from the group consisting of (i) amino acid residue F and (ii) amino acid residues that are conservative substitutions for (i). The antibody or antigen-binding fragment according to claim 1. [Claim 3] The antibody or antigen-binding fragment according to Claim 2, wherein X1 is amino acid residue D, X2 is selected from the group consisting of amino acid residues D and T, X3 is amino acid residue A, X10 is selected from the group consisting of amino acid residues G and D, X11 is selected from the group consisting of amino acid residues G and H, X15 is selected from the group consisting of amino acid residues R and E, X16 is selected from the group consisting of amino acid residues Q and K, X17 is selected from the group consisting of amino acid residues G and V, X22 is selected from the group consisting of amino acid residues S and Y, X23 is selected from the group consisting of amino acid residues E and Q, X27 is selected from the group consisting of amino acid residues R and H, X28 is selected from the group consisting of amino acid residues S and A, and X29 is amino acid residue F. [Claim 4] (1) The following three complementarity-determining regions (CDRs) of the heavy chain variable region (VH): (a) FTFX １ X ２ YX ３ VH CDR1 having the structure shown in MH (Sequence ID 76), (b) VIX ４ YX ５ X ６ X ７ X ８ KYYAX ９ VH CDR2 having the structure shown in SVKG (Sequence ID 77), (c) VH CDR3 having the structure shown in ARDTSRGHDI (Sequence ID 23) and / or (2) The following three complementarity-determining regions (CDRs) of the light chain variable region (VL): (d) X １２ ASQSVSX １３ X １４ VL CDR1 having the structure shown in YLA (Sequence ID 79), (e) GASX １８ X １９ X ２０ X ２１ VL CDR2 having the structure shown in (Sequence ID 8), (f) X ２４ X ２５ YX ２６ VL CDR3 having the structure shown in DYPPFT (Sequence ID 10) Includes, X １ However, (i) amino acid residues S, E, G and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２ However, (i) amino acid residues S and K and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ３ However, (i) an amino acid residue G and (ii) an amino acid residue that is a conservative substitution for (i) are selected from the group, X ４ However, (i) amino acid residues S and H and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ５ However, (i) amino acid residues E, H and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ６ However, (i) amino acid residues G, A and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ７ However, (i) amino acid residues S, E, T, V and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ８ However, (i) amino acid residues N, S, I, K and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ９ However, (i) amino acid residues D and E and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １２ However, (i) amino acid residues R and Q and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １３ However, (i) amino acid residues Y and S and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １４ However, (i) amino acid residues S, D, N and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X １８ is selected from the group consisting of (i) amino acid residues S, T, A and (ii) amino acid residues that are conservative substitutions for (i). X １９ However, (i) amino acid residues R, L and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２０ However, (i) amino acid residues N, A and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２１ However, (i) amino acid residues T and N and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２４ However, (i) amino acid residues S, G, Q and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２５ However, (i) amino acid residues E, Q, D and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group, X ２６ The antibody or antigen-binding fragment according to claim 1, wherein (i) amino acid residues A, S and (ii) amino acid residues that are conserved substitutions for (i) are selected from the group comprising (i) and (ii). [Claim 5] X1 is selected from the group consisting of amino acid residues S, E, and G; X2 is selected from the group consisting of amino acid residues S and K; X3 is amino acid residue G; X4 is selected from the group consisting of amino acid residues S and H; X5 is selected from the group consisting of amino acid residues E and H; X6 is selected from the group consisting of amino acid residues G and A; X7 is selected from the group consisting of amino acid residues S, E, T, and V; X8 is selected from the group consisting of amino acid residues N, S, I, and K; X9 is selected from the group consisting of amino acid residues D and E; X12 is selected from the group consisting of amino acid residues R and Q; X13 is selected from the group consisting of amino acid residues Y and S; X14 is selected from the group consisting of amino acid residues S, D, and N; X18 is selected from the group consisting of amino acid residues S, T, and A; X19 The antibody or antigen-binding fragment according to claim 4, wherein X 20 is selected from the group consisting of amino acid residues R and L, X 21 is selected from the group consisting of amino acid residues T and N, X 24 is selected from the group consisting of amino acid residues S, G and Q, X 25 is selected from the group consisting of amino acid residues E, Q and D, and X 26 is selected from the group consisting of amino acid residues A and S. [Claim 6] (a) VH CDR1 shown in Sequence ID No. 4 or 5, VH CDR2 shown in any one of Sequence ID Nos. 20, 19, or 21, and VH CDR3 shown in Sequence ID No. 24, VL CDR1 shown in any one of Sequence ID Nos. 42, 41, or 43, VL CDR2 shown in any one of Sequence ID Nos. 52, 51, or 53, and VL CDR3 shown in Sequence ID No. 59 or 58, or (b) VH CDR1 shown in any one of SEQ ID NOs: 1 to 3, VH CDR2 shown in any one of SEQ ID NOs: 12 to 18, 82, and VH CDR3 shown in SEQ ID NO: 23, VL CDR1 shown in any one of SEQ ID NOs: 37 to 40, VL CDR2 shown in any one of SEQ ID NOs: 45 to 50, and VL CDR3 shown in any one of SEQ ID NOs: 54 to 57 The antibody or antigen-binding fragment according to claim 1, comprising: [Claim 7] (1) VH CDR1 shown in Sequence ID 4, VH CDR2 shown in Sequence ID 20, and VH CDR3 shown in Sequence ID 24, VL CDR1 shown in Sequence ID 42, VL CDR2 shown in Sequence ID 52, and VL CDR3 shown in Sequence ID 59, (2) VH CDR1 shown in Sequence ID 1, VH CDR2 shown in Sequence ID 12, and VH CDR3 shown in Sequence ID 23, VL CDR1 shown in Sequence ID 37, VL CDR2 shown in Sequence ID 45, and VL CDR3 shown in Sequence ID 54, (3) VH CDR1 shown in Sequence ID 1, VH CDR2 shown in Sequence ID 13, and VH CDR3 shown in Sequence ID 23, VL CDR1 shown in Sequence ID 37, VL CDR2 shown in Sequence ID 45, and VL CDR3 shown in Sequence ID 54, (4) VH CDR1 shown in Sequence ID No. 2, VH CDR2 shown in Sequence ID No. 14, and VH CDR3 shown in Sequence ID No. 23, VL CDR1 shown in Sequence ID No. 38, VL CDR2 shown in Sequence ID No. 46, and VL CDR3 shown in Sequence ID No. 55, (5) VH CDR1 shown in Sequence ID No. 2, VH CDR2 shown in Sequence ID No. 15, and VH CDR3 shown in Sequence ID No. 23, VL CDR1 shown in Sequence ID No. 39, VL CDR2 shown in Sequence ID No. 47, and VL CDR3 shown in Sequence ID No. 56, (6) VH CDR1 shown in Sequence ID No. 2, VH CDR2 shown in Sequence ID No. 16, and VH CDR3 shown in Sequence ID No. 23, VL CDR1 shown in Sequence ID No. 39, VL CDR2 shown in Sequence ID No. 48, and VL CDR3 shown in Sequence ID No. 57, (7) VH CDR1 shown in Sequence ID No. 2, VH CDR2 shown in Sequence ID No. 17, and VH CDR3 shown in Sequence ID No. 23, VL CDR1 shown in Sequence ID No. 40, VL CDR2 shown in Sequence ID No. 49, and VL CDR3 shown in Sequence ID No. 57, (8) VH CDR1 shown in Sequence ID 3, VH CDR2 shown in Sequence ID 18, and VH CDR3 shown in Sequence ID 23, VL CDR1 shown in Sequence ID 38, VL CDR2 shown in Sequence ID 50, and VL CDR3 shown in Sequence ID 54, (9) VH CDR1 shown in Sequence ID No. 2, VH CDR2 shown in Sequence ID No. 82, and VH CDR3 shown in Sequence ID No. 23, VL CDR1 shown in Sequence ID No. 39, VL CDR2 shown in Sequence ID No. 48, and VL CDR3 shown in Sequence ID No. 57, (10) VH CDR1 shown in Sequence ID No. 4, VH CDR2 shown in Sequence ID No. 19, and VH CDR3 shown in Sequence ID No. 24, VL CDR1 shown in Sequence ID No. 41, VL CDR2 shown in Sequence ID No. 51, and VL CDR3 shown in Sequence ID No. 58, (11) VH CDR1 shown in Sequence ID No. 5, VH CDR2 shown in Sequence ID No. 21, and VH CDR3 shown in Sequence ID No. 24, VL CDR1 shown in Sequence ID No. 43, VL CDR2 shown in Sequence ID No. 53, and VL CDR3 shown in Sequence ID No. 59, or (12) The antibody or antigen-binding fragment according to claim 1, comprising VH CDR1 shown in SEQ ID NO: 6, VH CDR2 shown in SEQ ID NO: 22, and VH CDR3 shown in SEQ ID NO: 25, VL CDR1 shown in SEQ ID NO: 44, VL CDR2 shown in SEQ ID NO: 45, and VL CDR3 shown in SEQ ID NO:

60. [Claim 8] Further including the framework area of ​​human immunoglobulins, For example, it includes a framework region contained in the amino acid sequence encoded by a human germline antibody gene, and for example, it includes a heavy chain framework region contained in the amino acid sequence encoded by a human heavy chain germline gene and / or a light chain framework region contained in the amino acid sequence encoded by a human light chain germline gene. Optionally, the antibody or this antigen-binding fragment (1) Heavy chain variable region (VH) containing an amino acid sequence selected from the following: (i) The sequence shown in any one of sequence numbers 26-36 or 83, (ii) A sequence having one or more amino acid substitutions, deletions or additions (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) compared to any one of the sequences shown in SEQ ID NOs. 26-36, 83, or (iii) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to any one of the sequences described in sequence numbers 26-36, 83, and / or (2) Light chain variable region (VL) containing an amino acid sequence selected from the following: (i) The sequence shown in any one of sequence numbers 61 to 70, (ii) A sequence having one or more amino acid substitutions, deletions or additions (for example, substitutions, deletions or additions of 1, 2, 3, 4, or 5 amino acids) compared to any one of the sequences shown in Sequence ID No. 61 to 70, or (iii) A sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity compared to any one of the sequences described in sequence numbers 61 to 70. Includes, Preferably, (a) VH or a variant thereof containing the sequence shown in any one of sequence numbers 26-32 or 83, and VL or a variant thereof containing the sequence shown in any one of sequence numbers 61-66, or (b) VH or a variant thereof containing the sequence shown in any one of SEQ ID NOs. 33 to 35 and VL or a variant thereof containing the sequence shown in any one of SEQ ID NOs. 67 to 69 Includes, The variant has one or more amino acid substitutions, deletions, or additions (for example, 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions) compared to the derived sequence, or has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity, preferably the substitutions are conservative substitutions. For example, the sequence VH or a variant thereof, as shown in Sequence ID No. 26, and the sequence VL or a variant thereof, as shown in Sequence ID No. 61, For example, it includes VH or a variant thereof, which is the sequence shown in SEQ ID NO: 27, and VL or a variant thereof, which is the sequence shown in SEQ ID NO:

61. For example, including VH or a variant thereof, which is the sequence shown in SEQ ID NO: 28, and VL or a variant thereof, For example, including VH or a variant thereof, which is the sequence shown in SEQ ID NO: 29, and VL or a variant thereof, For example, the sequence VH or a variant thereof, as shown in Sequence ID No. 30, and the sequence VL or a variant thereof, as shown in Sequence ID No. 64, For example, it includes VH or a variant thereof, which is the sequence shown in SEQ ID NO: 31, and VL or a variant thereof, which is the sequence shown in SEQ ID NO:

65. For example, including VH or a variant thereof, which is the sequence shown in SEQ ID NO: 32, and VL or a variant thereof, For example, the sequence VH or a variant thereof, as shown in Sequence ID No. 33, and the sequence VL or a variant thereof, as shown in Sequence ID No. 67, For example, including VH or a variant thereof, which is the sequence shown in SEQ ID NO: 34, and VL or a variant thereof, For example, including VH or a variant thereof, which is the sequence shown in SEQ ID NO: 35, and VL or a variant thereof, For example, including VH or a variant thereof, which is the sequence shown in SEQ ID NO: 36, and VL or a variant thereof, For example, the sequence VH or a variant thereof, as shown in Sequence ID No. 83, and the sequence VL or a variant thereof, The antibody or antigen-binding fragment according to claim 1, wherein the variant has one or more amino acid substitutions, deletions or additions (for example, one, two, three, four or five amino acid substitutions, deletions or additions) compared to the derived sequence, or has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity, and preferably the substitutions are conservative substitutions. [Claim 9] (1) The antibody or the antigen-binding fragment further comprises a constant region derived from human immunoglobulin, (2) The heavy chain of the antibody or the antigen-binding fragment includes a constant region of the heavy chain derived from human immunoglobulin (e.g., IgG1, IgG2, IgG3, or IgG4), (3) The light chain of the antibody or this antigen-binding fragment contains a constant region of the light chain derived from human immunoglobulin (e.g., a constant region of the κ or λ chain), or (4) The antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab')2, Fv, disulfide-bonded Fv, scFv, diabody, and single-domain antibody (sdAb), and / or the antibody is a chimeric antibody, a bispecific antibody, or a polyspecific antibody. The antibody or antigen-binding fragment according to claim 1, having one or more features selected from the group consisting of the following. [Claim 10] An isolated nucleic acid molecule encoding the antibody according to any one of claims 1 to 9, or the antigen-binding fragment thereof, or the heavy chain variable region and / or light chain variable region. [Claim 11] A vector comprising the nucleic acid molecule described in claim 10, preferably a cloning vector or an expression vector. [Claim 12] A host cell comprising the nucleic acid molecule described in claim 10 or a vector containing the nucleic acid molecule. [Claim 13] A method for preparing an antibody or an antigen-binding fragment, comprising culturing the host cells described in claim 12 under conditions that enable the expression of the antibody or the antigen-binding fragment, and recovering the antibody or the antigen-binding fragment from the cell culture of the host cells. [Claim 14] A pharmaceutical composition comprising an antibody or antigen-binding fragment according to any one of claims 1 to 9, and a pharmaceutically acceptable carrier and / or excipient. [Claim 15] A pharmaceutical composition for activating CD40, enhancing immune cell activity, enhancing the immune response, and / or preventing and / or treating tumors or infections in a subject, comprising: (i) an antibody or antigen-binding fragment according to any one of claims 1 to 9; (ii) a nucleic acid molecule, vector or host cell comprising a nucleotide sequence encoding the antibody or antigen-binding fragment; or (iii) a pharmaceutical composition comprising the antibody or antigen-binding fragment and a pharmaceutically acceptable carrier and / or excipient, Preferably, the immune cells are T cells, B cells, DCs, macrophages, and / or NK cells. Preferably, the immune response is a CD40-mediated immune response. Preferably, the subject is a mammal, such as a human. Preferably, a pharmaceutical composition in which the antibody or the antigen-binding fragment is used alone or in combination with further pharmaceutically active substances. [Claim 16] A conjugate comprising an antibody or antigen-binding fragment according to any one of claims 1 to 9 and a detectable label that binds to the antibody or antigen-binding fragment, Preferably, the detectable label is a conjugate selected from the group consisting of enzymes (e.g., horseradish peroxidase or alkaline phosphatase), chemiluminescent reagents (e.g., acridine ester compounds, luminol and its derivatives or ruthenium derivatives), fluorescent dyes (e.g., fluorescein or fluorescent proteins), radionuclides, and biotin. [Claim 17] (i) an antibody or antigen-binding fragment according to any one of claims 1 to 9, or (ii) a conjugate comprising the antibody or antigen-binding fragment and a detectable label that binds thereto, Preferably, the conjugate is included, Preferably, the kit comprises an antibody or an antigen-binding fragment and a secondary antibody that specifically recognizes the antibody or antigen-binding fragment, wherein the secondary antibody may further comprise a detectable label, such as an enzyme (e.g., horseradish peroxidase or alkaline phosphatase), a chemiluminescent reagent (e.g., acridinium ester, luminol and its derivatives or ruthenium derivatives), a fluorescent dye (e.g., fluorescein or fluorescent protein), a radionuclide or biotin. [Claim 18] A method for detecting the presence or level of CD40 in a sample, comprising using (i) an antibody or antigen-binding fragment according to any one of claims 1 to 9, or (ii) a conjugate comprising the antibody or antigen-binding fragment and a detectable label bound thereto, Preferably, the assay is an immunoassay, such as an immunoblotting assay, an enzyme immunoassay (e.g., ELISA), a chemiluminescent immunoassay, a fluorescence immunoassay, or a radioimmunoassay. Preferably, this includes using the conjugate, Preferably, the method includes using an antibody or the antigen-binding fragment, and further includes detecting the antibody or the antigen-binding fragment using a secondary antibody possessing a detectable label [e.g., an enzyme (e.g., horseradish peroxidase or alkaline phosphatase), a chemiluminescent reagent (e.g., acridinium ester, luminol and its derivatives or ruthenium derivatives), a fluorescent dye (e.g., fluorescein or fluorescent protein), a radionuclide or biotin]. Preferably, the sample is a cell sample (e.g., immune cells) derived from a subject (e.g., a mammal, preferably a human or cynomolgus monkey). [Claim 19] (i) the antibody or antigen-binding fragment according to any one of claims 1 to 9 or (ii) the use of a conjugate comprising the antibody or antigen-binding fragment and a detectable label bound thereto, in the manufacture of a detection reagent for detecting the presence or level of CD40 in a sample using the detection reagent, Preferably, the sample used is a cell sample (e.g., immune cells) derived from the subject (e.g., a mammal, preferably a human or cynomolgus monkey).

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