Antibodies that specifically recognize CD40 and their use

Abstract

Provided are antibodies or antigen-binding fragments that specifically recognize CD40, as well as methods for their preparation and use.

Description

Cross-reference of related applications

[0001] This invention claims priority to the Chinese patent application “Antibody that specifically recognizes CD40 and its use,” application number 202110793032.1, filed on 14 July 2021, and the entirety of that application is incorporated herein by reference.

[0002] Citations for electronic sequence lists The contents of the electronic sequence listing (text name: CN_202106295612_SEQLIST.xml, recorded date: July 12, 2022, size: 141KB) are incorporated herein by reference in their entirety. [Technical Field]

[0003] The present invention provides an antibody or antigen-binding fragment that specifically recognizes CD40, as well as a method for preparing and using the same. [Background technology]

[0004] CD40 is one of the co-stimulatory members of the tumor necrosis factor receptor (TNFR) superfamily (Croft, Michael et al. Nature reviews. Drug discovery vol.12,2(2013):147-68). CD40's function was first identified in human B lymphocytes, where it is constitutively expressed, and CD40 signaling promotes B cell activation and proliferation (Clark, EA, and JA Ledbetter. Proceedings of the National Academy of Sciences of the United States of America vol.83,12(1986):4494-8). CD40, when expressed in other hematopoietic cells (e.g., monocytes and dendritic cells (DCs)), can promote cell survival and cytokine production (Stout, RD, and J Suttles. Immunology Today vol. 17, 10 (1996): 487-92), and also signals CD4, which expresses CD40L.+ It is known that CD40 signaling can induce T cell activation, proliferation, and cytokine production (Grewal, IS, and RA Flavell. Immunology Today vol.17,9(1996):410-4). CD40 signaling can also influence cellular responses in non-hematopoietic cells, including endothelial cells, epithelial cells, fibroblasts, and neurons (Hollenbaugh, D et al. The Journal of Experimental Medicine vol.182,1(1995):33-40; Karmann, K et al. Proceedings of the National Academy of Sciences of the United States of America vol.92,10(1995):4342-6; and Tan, Jun et al. The EMBO journal vol.21,4(2002):643-52).

[0005] CD40L is the first activated CD4 +It has been identified on the surface of T cells (Hollenbaugh, D et al. The EMBO journal vol.11,12(1992):4313-21), and the CD40-CD40L interaction between B cells and T cells indicates that CD40L is important in germinal center (GC) response and isotype class switching in response to T-dependent antigens (Kawabe, T et al. Immunity vol.1,3(1994):167-78). Furthermore, it has been demonstrated that CD40L can be expressed in a variety of cells, including mast cells, basophils, B cells, natural killer (NK) cells, macrophages, megakaryocytes, and platelets, revealing the broad role of the CD40 pathway in cell biology (Schonbeck, U, and P Libby. Cellular and molecular life sciences:CMLS vol.58,1(2001):4-43; and van Kooten, C, and J Banchereau. Journal of leukocyte biology vol.67,1(2000):2-17).

[0006] In many animal models, the CD40L / CD40 interaction is linked to systemic lupus erythematosus (Davidson, Anne et al. Annals of the New York Academy of Sciences vol.987(2003):188-98), experimental allergic encephalomyelitis (Toubi, Elias and Yehuda Shoenfeld. Autoimmunity vol.37,6-7(2004):457-64), collagen-induced arthritis (Durie, FH et al. Science (New York, NY) vol.261,5126(1993):1328-30), immune rejection (Goldwater, R et al. American journal of transplantation vol.13,4(2013):1040-1046), and atherosclerosis-mediated cardiovascular disease (Jansen, Matthijs F et al. Basic research in cardiology). vol.111,4(2016):38), Sjögren's syndrome (Okimura, K et al.American journal of transplantation vol.14,6(2014):1290-9), systemic sclerosis (Boleto, Goncalo et al.Frontiers in immunology vol.9) 2998.18Dec.2018), immune thrombocytopenic purpura (Kuwana,Masataka et al.Blood vol.103,4(2004):1229-36), inflammatory bowel disease (Senhaji,Nezha et al.Frontiers in immunology vol.6 529.16Oct.2015), diabetes (Balasa,B et al. al.Journal of Immunology (Baltimore, Md.: 1950) vol.159,9(1997):4620-7), Rheumatoid Arthritis (RA) (Gotoh, Haruhiko et al. The Journal of Rheumatology vol.31,8(2004):1506-12), and Cancer (Li, Rui et al. Molecular Biology Reports vol.It has been demonstrated to be associated with many diseases, including those listed in 39,9(2012):8741-7; and Li, Rui et al. (Immunology Letters vol.131,2(2010):120-5).

[0007] Therefore, neutralizing the binding of CD40 to CD40L may be a therapeutic approach for diseases and conditions mediated by the CD40 / CD40L pathway. Patent application WO200563981A1 discloses the anti-human CD40 antibody ASKP1240 (used as a control antibody in the embodiment portion of the present invention), and patent application WO201265950A1 discloses the anti-human CD40 antibody CFZ533 (used as a control antibody in the embodiment portion of the present invention).

[0008] All publications, patents, patent applications, and the information disclosed in published patent applications referred to herein are incorporated herein by reference in their entirety. [Overview of the Initiative]

[0009] In some examples, the isolated anti-CD40 antibody had a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V Hcomprises heavy chain complementarity determining region (HC-CDR) 1, HC-CDR2, and HC-CDR3, wherein the heavy chain complementarity determining region (HC-CDR) 1 comprises GYX1VN (SEQ ID NO:81), X1 is L, T or V, the HC-CDR2 comprises X1IX2PX3X4GX5TX6YNQKFKG (SEQ ID NO:82), X1 is L or M, X2 is A, N or V, X3 is E, K, L, M, R, S, T or Y, X4 is H, N, S or T, X5 is G or R, X6 is A, S, T or W, the HC-CDR3 comprises SX1X2X3X4PYAX5DY (SEQ ID NO:83), X1 is G, I, L, M, P, R, S or Y, X2 is W or Y, X3 is S or V, X4 is D, K, L, N, R, T or V, X5 is A, G, M, N, Q or S, said V L comprises light chain complementarity determining region (LC-CDR) 1, LC-CDR2, and LC-CDR3, wherein the light chain complementarity determining region (LC-CDR) 1 comprises KASQNVHTX1VA (SEQ ID NO:84), X1 is A, H, N or R, the LC-CDR2 comprises LX1SDRRT (SEQ ID NO:85), X1 is A or R, the LC-CDR3 comprises LQX1WX2X3PLT (SEQ ID NO:86), X1 is A or H, X2 is N, S or T, X3 is A, Q, R, S or Y, and is any of the isolated anti-CD40 antibodies as described above.

[0010] In some embodiments, V H and V L comprises, said V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs:1-3 or a variant of the amino acid sequence shown in any of SEQ ID NOs:1-3 including at most about three amino acid substitutions, HC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs:9-26 or a variant of the amino acid sequence shown in any of SEQ ID NOs:9-26 including at most about three amino acid substitutions, and HC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs:32-52 or a variant of the amino acid sequence shown in any of SEQ ID NOs:32-52 including at most about three amino acid substitutions, and V L The present invention provides an isolated anti-CD40 antibody comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs: 58-61 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-61 containing at least approximately three amino acid substitutions, LC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs: 65-66 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 65-66 containing at least approximately three amino acid substitutions, and LC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs: 70-77 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 70-77 containing at least approximately three amino acid substitutions.

[0011] In some embodiments, V H and V L Including the above V H This refers to V, which is represented by one of the amino acid sequences of SEQ ID NOs: 87-119. H The V L This refers to V, which is represented by one of the amino acid sequences of SEQ ID NOs:126-138. LThis invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0012] In some embodiments, (i)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:87. H The V L This is the V shown in amino acid sequence SEQ ID NO:126. L (ii)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:88. H The V L This is the V shown in amino acid sequence SEQ ID NO:127. L (iii)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:89. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L (iv)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:94. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L (v)V Hand V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:96. H The V L This is the V shown in amino acid sequence SEQ ID NO:132. L (vi)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:101. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L (vii)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:110. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L (viii)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:114. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L (ix)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:115. H The VL This is the V shown in amino acid sequence SEQ ID NO:129. L (x)V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:117. H The V L This is the V shown in amino acid sequence SEQ ID NO:129. L This invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0013] In some embodiments, (i)V H Or V H mutants of V L Or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:9, and HC-CDR3 comprises amino acid sequence SEQ ID NO:32, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 58, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions; (ii)V H Or V H mutants of V L Or V L Includes variants of the V Hcomprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein said HC-CDR1 comprises the amino acid sequence SEQ ID NO:1, said HC-CDR2 comprises the amino acid sequence SEQ ID NO:10, said HC-CDR3 comprises the amino acid sequence SEQ ID NO:32, and the V H of the variant of HC-CDRs comprises at most about 5 amino acid substitutions, and the V L comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein said LC-CDR1 comprises the amino acid sequence SEQ ID NO:59, said LC-CDR2 comprises the amino acid sequence SEQ ID NO:65, said LC-CDR3 comprises the amino acid sequence SEQ ID NO:70, and the V L of the variant of LC-CDRs comprises at most about 5 amino acid substitutions; (iii) V H or the variant of said V H and V L or the variant of said V L comprises the variant of V H comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein said HC-CDR1 comprises the amino acid sequence SEQ ID NO:1, said HC-CDR2 comprises the amino acid sequence SEQ ID NO:13, said HC-CDR3 comprises the amino acid sequence SEQ ID NO:34, and the V H of the variant of HC-CDRs comprises at most about 5 amino acid substitutions, and the V L comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein said LC-CDR1 comprises the amino acid sequence SEQ ID NO:59, said LC-CDR2 comprises the amino acid sequence SEQ ID NO:65, said LC-CDR3 comprises the amino acid sequence SEQ ID NO:70, and the V L of the variant of LC-CDRs comprises at most about 5 amino acid substitutions; (iv) V H or the variant of said V H and V L or the variant of said V L comprises the variant of V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:37, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 73. L The LC-CDRs of the mutants contain at most about five amino acid substitutions; (v)V H Or V H mutants of V L Or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:20, and HC-CDR3 comprises amino acid sequence SEQ ID NO:42, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions; (vi)V H Or V H mutants of V L Or V L Includes variants of the V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:14, and HC-CDR3 comprises amino acid sequence SEQ ID NO:43. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions; (vii)V H Or V H mutants of V L Or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:51. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions; (viii)V H Or V H mutants of V L Or V L Includes variants of the V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:11, and HC-CDR3 comprises amino acid sequence SEQ ID NO:48. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 71, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions; or (ix)V H Or V H mutants of V L Or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:18, and HC-CDR3 comprises amino acid sequence SEQ ID NO:47, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 71, and V L The LC-CDR variants provide isolated anti-CD40 antibodies containing at most about five amino acid substitutions.

[0014] In some examples, the isolated anti-CD40 antibody was V H and V L Including the above V HThis includes an amino acid sequence shown in any of SEQ ID NOs:87-119 or a mutant having at least approximately 90% sequence identity with an amino acid sequence shown in any of SEQ ID NOs:87-119, and the V L This refers to any of the isolated anti-CD40 antibodies described above, which include the amino acid sequence shown in any of SEQ ID NOs:126-138 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in any of SEQ ID NOs:126-138.

[0015] In some embodiments, (i)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:87, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:87, and the V L (ii)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:88, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:88, and the V L (iii)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:89, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:89, and the V L (iv)V H and V L Including the above V HThis includes amino acid sequence SEQ ID NO:94, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:94, and the V L This includes amino acid sequence SEQ ID NO:128, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:128; (v)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:96, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:96, and the V L (vi)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:101, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:101, and the V L (vii)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:110, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:110, and the V L (viii)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:114, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:114, and the V LThis includes amino acid sequence SEQ ID NO:128, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:128; (ix)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:115, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:115, and the V L This includes amino acid sequence SEQ ID NO:129, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:129; or (x)V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:117, or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:117, and the V L This provides isolated anti-CD40 antibodies containing amino acid sequence SEQ ID NO:129, or variants having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:129.

[0016] In some embodiments, V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:120. H The V L This is the V shown in amino acid sequence SEQ ID NO:139. L This invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0017] In some embodiments, V H or V H mutants of V L or V L Includes variants of the V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:4, HC-CDR2 comprises amino acid sequence SEQ ID NO:27, and HC-CDR3 comprises amino acid sequence SEQ ID NO:53. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:67, and LC-CDR3 comprises amino acid sequence SEQ ID NO:78. L The LC-CDR variants provide isolated anti-CD40 antibodies containing at most about five amino acid substitutions.

[0018] In some examples, the isolated anti-CD40 antibody was V H and V L Including the above V H This includes the amino acid sequence shown in SEQ ID NO:120 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:120, and the V L This refers to an isolated anti-CD40 antibody as described above, containing either the amino acid sequence shown in SEQ ID NO:139 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:139.

[0019] In some embodiments, V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:121. H The V L This is the V shown in amino acid sequence SEQ ID NO:140. L This invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0020] In some embodiments, V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO: 5, HC-CDR2 comprises amino acid sequence SEQ ID NO: 27, and HC-CDR3 comprises amino acid sequence SEQ ID NO: 54. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:67, and LC-CDR3 comprises amino acid sequence SEQ ID NO:78. L The LC-CDR variants provide isolated anti-CD40 antibodies containing at most about five amino acid substitutions.

[0021] In some embodiments, V H and V L Including the above V H This includes the amino acid sequence shown in SEQ ID NO:121 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:121, and the V L This refers to any of the isolated anti-CD40 antibodies described above, including the amino acid sequence shown in SEQ ID NO:140 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:140.

[0022] In some embodiments, V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:122. HThe V L This is the V shown in amino acid sequence SEQ ID NO:141. L This invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0023] In some embodiments, V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:6, HC-CDR2 comprises amino acid sequence SEQ ID NO:28, and HC-CDR3 comprises amino acid sequence SEQ ID NO:55, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:63, LC-CDR2 comprises amino acid sequence SEQ ID NO:68, and LC-CDR3 comprises amino acid sequence SEQ ID NO:79, and V L The LC-CDR variants provide isolated anti-CD40 antibodies containing at most about five amino acid substitutions.

[0024] In some embodiments, V H and V L Including the above V H This includes the amino acid sequence shown in SEQ ID NO:122 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:122, and the V L This refers to any of the isolated anti-CD40 antibodies described above, including the amino acid sequence shown in SEQ ID NO:141 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:141.

[0025] In some embodiments, V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:123. H The V L This is the V shown in amino acid sequence SEQ ID NO:142. L This invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0026] In some embodiments, V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:7, HC-CDR2 comprises amino acid sequence SEQ ID NO:29, and HC-CDR3 comprises amino acid sequence SEQ ID NO:56. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO:64, LC-CDR2 includes amino acid sequence SEQ ID NO:69, and LC-CDR3 includes amino acid sequence SEQ ID NO:80, and V L The LC-CDR variants provide isolated anti-CD40 antibodies containing at most about five amino acid substitutions.

[0027] In some embodiments, V H and V L Including the above V HThis includes the amino acid sequence shown in SEQ ID NO:123 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:123, and the V L This refers to any of the isolated anti-CD40 antibodies described above, including the amino acid sequence shown in SEQ ID NO:142 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:142.

[0028] In some embodiments, V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:124. H The V L This is the V shown in amino acid sequence SEQ ID NO:143. L This invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0029] In some embodiments, V H or V H mutants of V L or V L Includes variants of the V H This includes HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 includes amino acid sequence SEQ ID NO:8, HC-CDR2 includes amino acid sequence SEQ ID NO:30, and HC-CDR3 includes amino acid sequence SEQ ID NO:57, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:67, and LC-CDR3 comprises amino acid sequence SEQ ID NO:80, and V LThe LC-CDR variants provide isolated anti-CD40 antibodies containing at most about five amino acid substitutions.

[0030] In some embodiments, V H and V L Including the above V H This includes the amino acid sequence shown in SEQ ID NO:124 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:124, and the V L This refers to any of the isolated anti-CD40 antibodies described above, including the amino acid sequence shown in SEQ ID NO:143 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:143.

[0031] In some embodiments, V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:125. H The V L This is the V shown in amino acid sequence SEQ ID NO:144. L This invention provides isolated anti-CD40 antibodies containing LC-CDR1, LC-CDR2, and LC-CDR3.

[0032] In some embodiments, V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:4, HC-CDR2 comprises amino acid sequence SEQ ID NO:31, and HC-CDR3 comprises amino acid sequence SEQ ID NO:53, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V LIt comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:68, and LC-CDR3 comprises amino acid sequence SEQ ID NO:79, and V L The LC-CDR variants provide isolated anti-CD40 antibodies containing at most about five amino acid substitutions.

[0033] In some embodiments, V H and V L Including the above V H This includes the amino acid sequence shown in SEQ ID NO:125 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:125, and the V L This refers to any of the isolated anti-CD40 antibodies described above, including the amino acid sequence shown in SEQ ID NO:144 or a variant having at least approximately 90% sequence identity with the amino acid sequence shown in SEQ ID NO:144.

[0034] In several examples, the Kd values ​​for binding between isolated anti-CD40 antibodies and human CD40 were 0.1 pM to 1 nM.

[0035] In some examples, isolated anti-CD40 antibodies are provided that specifically bind to CD40 competitively with any of the isolated anti-CD40 antibodies described above. In some examples, isolated anti-CD40 antibodies are provided that specifically bind to the same epitope as any of the isolated anti-CD40 antibodies described above.

[0036] In some examples, the isolated anti-CD40 antibody is one of the above-described isolates containing an Fc fragment. In some examples, the isolated anti-CD40 antibody is a full-length IgG antibody. In some examples, the isolated anti-CD40 antibody is a full-length IgG1, IgG2, IgG3, or IgG4 antibody. In some examples, the isolated anti-CD40 antibody is a chimeric antibody, a fully human antibody, or a humanized antibody. In some examples, the isolated anti-CD40 antibody is an antigen-binding fragment, which is selected from Fab, Fab', F(ab)'2, Fab'-SH, single-chain antibody (scFv), Fv fragment, dAb, Fd, nanobody, diabody, and linear antibody.

[0037] In some examples, isolated nucleic acid molecules are provided, the nucleic acid molecules encoding any of the above-described anti-CD40 antibodies. In some examples, vectors comprising any of the above-described nucleic acid molecules are provided. In some examples, host cells comprising any of the above-described anti-CD40 antibodies, any of the above-described nucleic acid molecules, or any of the above-described vectors are provided. In some examples, a method for preparing an anti-CD40 antibody is provided, comprising a) culturing any of the above-described host cells under conditions in which the anti-CD40 antibody can be effectively expressed; and b) obtaining the expressed anti-CD40 antibody from the host cells.

[0038] In some embodiments, a method for treating a disease or condition of a target subject is provided, comprising administering an effective amount of any of the above-described anti-CD40 antibodies to the target subject. In some embodiments, the use of any of the above-described anti-CD40 antibodies in the preparation of a drug composition for treating a disease or condition of a target subject is provided. In some embodiments, the use of any of the above-described anti-CD40 antibodies or a drug composition containing an anti-CD40 antibody in the preparation of a drug for treating a disease or condition is provided. In some embodiments, the disease or condition is related to the CD40 signaling pathway and includes autoimmune diseases and / or inflammatory diseases and / or cancer and / or transplant-related diseases or conditions.In some embodiments, the disease or condition is, for example, systemic lupus erythematosus (SLE), discoid lupus erythematosus, lupus nephritis, inflammatory arthritis, hyperacute rejection, acute or chronic rejection, and / or graft-versus-host disease, organ or tissue transplant rejection, multiple sclerosis, hyper-IgE syndrome, polyarteritis nodosa, primary biliary cirrhosis, inflammatory bowel disease, Crohn's disease, Graves' disease (gluten-sensitive bowel syndrome), autoimmune hepatitis, pernicious anemia, autoimmune hemolytic anemia, scleroderma, myasthenia gravis, idiopathic thrombocytopenic purpura, autoimmune thyroiditis, Graves' disease, Hashimoto's thyroiditis Thyroiditis, immune complex disease, chronic fatigue-related immune dysfunction syndrome (CFIDS), polymyositis, dermatomyositis, cryoglobulinemia, thrombolysis, cardiomyopathy, pemphigus vulgaris, interstitial pulmonary fibrosis, type 1 and type 2 diabetes, type 1, type 2, type 3 and type 4 delayed-type hypersensitivity, allergies or allergic diseases, unnecessary / unintended immune responses to therapeutic proteins (e.g., factor VII of hemophilia), asthma, Rich-Strauss syndrome (allergic granulomatosis), atopic dermatitis, allergic contact dermatitis and irritant contact dermatitis, urticaria, IgE-mediated allergy, atherosclerosis, vasculitis, idiopathic inflammatory muscle disease, hemolytic disease, Alzheimer's disease, chronic inflammatory demyelinating polyneuropathy, Sjögren's syndrome The following are selected from inflammatory arthritis, psoriasis, hidradenitis suppurativa, urethritis suppurativa, sarcoidosis, ulcerative colitis, tumors, nephritis, respiratory distress syndrome, epididymitis, glomerulonephritis, and hyperthyroidism; the inflammatory arthritis includes, but is not limited to, juvenile arthritis, rheumatoid arthritis, psoriatic arthritis, Reiter's syndrome, ankylosing spondylitis, and gouty arthritis; and the tumors include, but are not limited to, non-Hodgkin lymphoma, chronic lymphocytic leukemia, multiple myeloma, B-cell lymphoma, high-grade / intermediate-grade / low-grade B-cell lymphoma, acute B-cell lymphoblastic leukemia, granulocytic leukemia, and Hodgkin's disease.

[0039] Simultaneously, we provide pharmaceutical compositions, reagent kits, and products containing any of the anti-CD40 antibodies described above. [Brief explanation of the drawing]

[0040] [Figure 1] The results shown in Figure 1A indicate that anti-CD40 antibodies D1, D6, or D7 inhibited human CD40L-induced CD95 expression in Ramos cells compared to the control antibody ASKP1240. The results shown in Figure 1B indicate that, after crosslinking with an anti-immunoglobulin antibody, anti-CD40 antibodies D1, D7, and the control antibody ASKP1240 had little to no side effect of activating the CD40 pathway, while the control antibody CFZ533 showed some agonist activity. [Figure 2] The results shown in Figure 2 indicate that, compared to the control antibody ASKP1240, anti-CD40 antibodies D1, D6, or D7 inhibited CD40L-induced CD23 expression in PBMCs. [Figure 3] The results shown in Figures 3A-3B indicate that anti-CD40 antibodies D1, D6, or D7 inhibited the secretion of CD40L-induced cytokines in iDCs compared to the control antibody ASKP1240. The results shown in Figure 3A indicate that anti-CD40 antibodies D1, D6, or D7 inhibited the secretion of CD40L-induced IL12p40 in iDCs. The results shown in Figure 3B indicate that anti-CD40 antibodies D1, D6, or D7 inhibited the secretion of CD40L-induced TNF-α in iDCs. [Figure 4] The results shown in Figure 4 demonstrate that, compared to the control antibody ASKP1240, the optimized anti-CD40 antibodies D21, D26, D33, D42, D47, and D49 inhibited the activation of the NF-κB signaling pathway induced by human CD40L. [Figure 5]The results shown in Figures 5A-5B demonstrate the binding affinity of the optimized anti-CD40 antibodies to different species of CD40 compared to the control antibody ASKP1240. Figure 5A shows the binding curves of the optimized anti-CD40 antibodies D21, D26, D33, D42, D47, and D49 to human CD40. Figure 5B shows the binding curves of the optimized anti-CD40 antibodies D21, D26, D33, D42, D47, and D49 to rhesus monkey CD40. [Figure 6] The results shown in Figure 6 demonstrate that, compared to the control antibody ASKP1240, the optimized anti-CD40 antibodies D21, D26, D33, D42, D47, and D49 inhibited human CD40L-induced CD95 expression in Ramos cells. [Figure 7] The results shown in Figure 7 demonstrate that, compared to the control antibody ASKP1240, the optimized anti-CD40 antibodies D33, D42, and D47 inhibited CD40L-induced CD23 expression in PBMCs. [Figure 8] The results shown in Figures 8A-8D demonstrate the blocking effect of anti-CD40 antibodies on the T cell-dependent antibody response compared to the control antibody ASKP1240. Figure 8A shows that optimized anti-CD40 antibodies D33, D42, or D47 nearly completely blocked the production of MASP2 antigen-specific IgG at a dose of 3 mpk. Figure 8B shows that optimized anti-CD40 antibodies D33, D42, or D47 nearly completely blocked the production of MASP2 antigen-specific IgM at a dose of 3 mpk. Figure 8C shows that optimized anti-CD40 antibodies D33, D42, or D47 nearly completely blocked the production of KLH antigen-specific IgG at a dose of 1 mpk. Figure 8D shows that optimized anti-CD40 antibodies D33, D42, or D47 nearly completely blocked the production of KLH antigen-specific IgM at a dose of 1 mpk. [Figure 9] The results shown in Figure 9 are the drug serum concentration curves in rats for the optimized anti-CD40 antibodies D33, D42, or D47 compared to the control antibody ASKP1240. [Modes for carrying out the invention]

[0041] According to one aspect of the present invention, an anti-CD40 antibody molecule is provided. By combining hybridoma technology, scFv phage library screening, humanization technology, affinity maturation, and appropriately designed biochemical and biological experiments, an effective antibody molecule capable of binding to human CD40 and inhibiting the action of human CD40 and its receptor has been identified. Results herein show that the antibody described herein binds to human CD40 and / or monkey CD40 with high affinity compared to the known anti-CD40 antibody ASKP1240, and, surprisingly, has been proven to be even more effective than ASKP1240 in various biological experiments.

[0042] The anti-CD40 antibodies provided by the present invention include, for example, full-length anti-CD40 antibodies, anti-CD40 single-chain antibodies (scFvs), anti-CD40Fc fusion proteins, multispecific (e.g., bispecific) anti-CD40 antibodies, and anti-CD40 immunoconjugates.

[0043] According to one embodiment, the present invention provides an anti-CD40 antibody, wherein the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V HThis includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes GYX1VN (SEQ ID NO: 81), where X1 is L, T, or V; the HC-CDR 2 includes X1IX2PX3X4GX5TX6YNQKFKG (SEQ ID NO: 82), where X1 is L or M, X2 is A, N, or V, X3 is E, K, L, M, R, S, T, or Y, X4 is H, N, S, or T, X5 is G or R, X6 is A, S, T, or W; and the HC-CDR 3 includes SX1X2X3X4PYAX5DY (SEQ ID NO: 82). NO:83) includes, where X1 is G, I, L, M, P, R, S, or Y, X2 is W, or Y, X3 is S, or V, X4 is D, K, L, N, R, T, or V, and X5 is A, G, M, N, Q, or S, and the V L This comprises a light chain complementarity determination region (LC-CDR) 1, an LC-CDR 2, and an LC-CDR 3, wherein the light chain complementarity determination region (LC-CDR) 1 includes KASQNVHTX1VA (SEQ ID NO: 84), where X1 is A, H, N, or R; the LC-CDR 2 includes LX1SDRRT (SEQ ID NO: 85), where X1 is A or R; the LC-CDR 3 includes LQX1WX2X3PLT (SEQ ID NO: 86), where X1 is A or H, X2 is N, S, or T, and X3 is A, Q, R, S, or Y.

[0044] According to one embodiment, the present invention provides an anti-CD40 antibody, wherein the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes DAYMH (SEQ ID NO: 4), the HC-CDR 2 includes RIDPANGNTKYDPKFQG (SEQ ID NO: 27), and the HC-CDR 3 includes NYYGSRYPFAY (SEQ ID NO: 53), and the VL This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes SASSSVSYMH (SEQ ID NO: 62), LC-CDR2 includes STSNLAS (SEQ ID NO: 67), and LC-CDR3 includes QQGSSYPLT (SEQ ID NO: 78).

[0045] According to one embodiment, the present invention provides an anti-CD40 antibody, wherein the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes DSYMH (SEQ ID NO: 5), the HC-CDR 2 includes RIDPANGNTKYDPKFQG (SEQ ID NO: 27), and the HC-CDR 3 includes NYYGAAYPFAY (SEQ ID NO: 54), and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes SASSSVSYMH (SEQ ID NO: 62), LC-CDR2 includes STSNLAS (SEQ ID NO: 67), and LC-CDR3 includes QQGSSYPLT (SEQ ID NO: 78).

[0046] According to one embodiment, the present invention provides an anti-CD40 antibody, wherein the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes DSYIH (SEQ ID NO: 6), the HC-CDR 2 includes RIDPTNGNTNFDPKFRG (SEQ ID NO: 28), and the HC-CDR 3 includes NYYGSRYPFSY (SEQ ID NO: 55), and the V LThis includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes SGSSSVSYMH (SEQ ID NO: 63), LC-CDR2 includes FTSNLAS (SEQ ID NO: 68), and LC-CDR3 includes QQGSTYPLT (SEQ ID NO: 79).

[0047] According to one embodiment, the present invention provides an anti-CD40 antibody, wherein the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H It includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes DHGMH (SEQ ID NO: 7), the HC-CDR 2 includes VISTSYGVATYNQKFKG (SEQ ID NO: 29), and the HC-CDR 3 includes RDSKSPFDY (SEQ ID NO: 56), and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes SASSSISSNYLH (SEQ ID NO: 64), LC-CDR2 includes RTSNLAS (SEQ ID NO: 69), and LC-CDR3 includes HQYHRSPLT (SEQ ID NO: 80).

[0048] According to one embodiment, the present invention provides an anti-CD40 antibody, wherein the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes DHAIH (SEQ ID NO: 8), the HC-CDR 2 includes AISTNYGAATYNQKFKG (SEQ ID NO: 30), and the HC-CDR 3 includes RGSHGDWFAY (SEQ ID NO: 57), and the V LThis includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes SASSSVSYMH (SEQ ID NO: 62), LC-CDR2 includes STSNLAS (SEQ ID NO: 67), and LC-CDR3 includes HQYHRSPLT (SEQ ID NO: 80).

[0049] In another aspect, the present invention provides an anti-CD40 antibody, the anti-CD40 antibody having a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes DAYMH (SEQ ID NO: 4), the HC-CDR 2 includes RIDPANGYTKSDPKFQG (SEQ ID NO: 31), and the HC-CDR 3 includes NYYGSRYPFAY (SEQ ID NO: 53), and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes SASSSVSYMH (SEQ ID NO: 62), LC-CDR2 includes FTSNLAS (SEQ ID NO: 68), and LC-CDR3 includes QQGSTYPLT (SEQ ID NO: 79).

[0050] Simultaneously, the present invention also provides nucleic acids encoding anti-CD40 antibodies, compositions containing anti-CD40 antibodies, and methods for producing and using anti-CD40 antibodies.

[0051] definition As described herein, “treatment” or “treating” is a method for obtaining a favorable or desired outcome, including clinical outcomes. In view of the object of the present invention, such favorable or desired clinical outcomes include, but are not limited to, one or more of the following: relief of one or more symptoms of the disease, reduction of the severity of the disease, stabilization of the disease (e.g., prevention or delay of disease exacerbation), prevention or delay of disease spread (e.g., metastasis), prevention or delay of disease recurrence, delay or slowing of disease progression, improvement of the disease state, relief of the disease (in part or in whole), reduction of the dose of one or more other agents necessary for the treatment of the disease, delay of disease progression, improvement or enhancement of quality of life, weight gain, and / or extension of survival. Also, “treatment” includes a reduction of the pathological outcome of the disease (e.g., in the case of cancer, tumor volume). The methods of the present invention take into consideration one or more of these treatments.

[0052] The term "antibody" includes full-length antibodies and their antigen-binding fragments. A full-length antibody comprises two heavy chains and two light chains. The variable regions of the light and heavy chains are responsible for antigen binding. The variable regions in the two chains generally contain three hypervariable loops called complementarity-determining regions (CDRs) (light chain (LC) CDRs include LC-CDR1, LC-CDR2, and LC-CDR3, and heavy chain (HC) CDRs include HC-CDR1, HC-CDR2, and HC-CDR3). The CDR boundaries of the antibodies or antigen-binding fragments disclosed herein can be defined or recognized according to the conventions of Kabat, Chothia, or Al-Lazikani (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). Three CDR regions of the heavy or light chain are inserted between flanking regions called framework regions (FRs), which are more conserved than the CDR regions and form a scaffold supporting the hypervariable loop. The constant regions of the heavy and light chains do not participate in antigen binding but exhibit various effects and functions. Antibodies are classified based on the amino acid sequence of their heavy chain constant regions. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, characterized by having α, δ, ε, γ, and μ type heavy chains, respectively. Multiple major antibody classes are divided into subclasses, for example, IgG1 (γ1 heavy chain), IgG2 (γ2 heavy chain), IgG3 (γ3 heavy chain), IgG4 (γ4 heavy chain), IgA1 (α1 heavy chain), or IgA2 (α2 heavy chain).

[0053] As described herein, the term “antigen-binding fragment” includes, for example, antibody fragments such as diabody, Fab, Fab', F(ab')2, Fv fragment, disulfide-stabilized Fv fragment (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized diabody (dsdiabody), single-stranded Fv (scFv), scFv dimer (bivalent diabody), multispecific antibodies consisting of antibody fragments containing one or more CDRs, single-domain antibodies, nanobody, domain antibodies, bivalent domain antibodies, or any other antibody fragments that can bind to an antigen but do not contain a complete antibody structure. Antigen-binding fragments can bind to the same antigen as a parent antibody or parent antibody fragment (e.g., parent scFv). Antigen-binding fragments also include fusion proteins containing the above antibody fragments. In some examples, the antigen-binding fragment may comprise one or more CDRs derived from a specific human antibody, which were implanted into a framework region derived from one or more different human antibodies.

[0054] As described herein, the term “epitope” means a specific group of atoms or amino acids on an antigen to which an antibody or antibody moiety binds. When two antibodies or antibody moieties indicate that they competitively bind to a particular antigen, they may bind to the same epitope on the antigen.

[0055] As used herein, if the first antibody inhibits the binding of the second antibody to the CD40 target by at least 50% (e.g., at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) at equimolar concentrations, then the first antibody “competes” with the second antibody to bind to the CD40 target, and vice versa. PCT publication WO03 / 48731 describes a “epitope classification” method for high-throughput antibodies based on cross-competition.

[0056] As described herein, the terms “specifically bind,” “specifically recognize,” or “specific to…” mean a measurable and reproducible interaction, such as the binding of an antibody to a target which can confirm the presence of that target in a heterogeneous population of cells containing biomolecules. For example, the ability of an antibody to specifically recognize a target (which may be an epitope) means that the binding of that antibody to that target has a higher affinity, binding strength, is easier, and / or more persistent compared to binding to other targets. In some examples, an antibody that specifically recognizes an antigen reacts with one or more antigenic determinants of the antigen, and its binding affinity is at least 10 times that of binding to other targets.

[0057] As described herein, “isolated” anti-CD40 antibody means an anti-CD40 antibody that (1) is unrelated to a naturally occurring protein, (2) does not contain other proteins of the same origin, (3) is expressed by cells of a different species, or (4) does not exist in nature.

[0058] As used herein, the term “isolated nucleic acid” means a genome, cDNA, or a combination of synthetic nucleic acids or other nucleic acids. Depending on its origin, such “isolated nucleic acid” means (1) unrelated to all or part of polynucleotides in “isolated nucleic acid” found in nature, (2) operably ligable with polynucleotides not naturally ligated to it, or (3) not existing in nature as part of a longer sequence.

[0059] As used herein, the term “CDR” or “complementarity-determining region” refers to discontinuous antigen-binding sites found within the variable domains of heavy and light chain polypeptides. ReferencesKabat et al., J.Biol.Chem.252:6609-6616(1977), Kabat et al.,USDept.of Health and Human Services, “Sequences of proteins of immunological interest”(1991);Chothia et al.,J.Mol.Biol.196:901-917(1987);Al-Lazikani B.et al.,J.Mol.Biol.,273:927-948(1997);MacCallum et al.,J.Mol.Biol.262:732-745(1996);Abhinandan and Martin,Mol.Immunol.,45:3832-3839(2008);Lefranc MPet al.,Dev.Comp.Immunol.,27:55-77(2003); and Honegger These specific regions are described in and Pluckthun, J.Mol.Biol., 309:657-670 (2001), and when comparing these specific regions with one another, these definitions include overlapping or subsets of amino acid residues. However, using any of the definitions to refer to the CDR of an antibody or grafted antibody or its variants falls within the scope of the terms defined and used herein. Table 1 shows, for comparison, the positions of amino acid residues contained in the CDRs defined by each of the references cited above. The prediction algorithms and bonding interfaces for CDR are known in the art, including, for example, those described in Abhinandan and Martin, Mol.Immunol., 45:3832-3839 (2008); Ehrenmann F. et al., Nucleic Acids Res., 38:D301-D307 (2010); and Adolf-Bryfogle J. et al., Nucleic Acids Res., 43:D432-D438 (2015).The contents of the references cited in this paragraph are incorporated herein by reference in their entirety for use in the present invention and in one or more claims contained herein.

[0060] [Table 1]

[0061] The term "chimeric antibody" means an antibody in which a portion of the heavy chain and / or light chain is derived from a particular species, or matches or is homologous to a corresponding sequence in an antibody belonging to a particular antibody type or subclass, and the remainder of these chains is match or is homologous to a corresponding sequence in an antibody derived from another species, or an antibody belonging to another antibody type or subclass, and also means a fragment of such an antibody insofar as it has the biological activity of the present invention (see US Patent No. 4816567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).

[0062] "Fv" is the smallest antibody fragment containing a complete antigen recognition and binding site. This fragment is a dimer formed by a tight non-covalent bond between one heavy-chain variable domain and one light-chain variable domain. The folding of these two domains induces six hypervariable loops (three rings each in the light and heavy chains) that provide the antibody with amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of the Fv fragment containing only three antigen-specific CDRs) has the ability to recognize and bind to antigens, despite having lower affinity than the complete binding site.

[0063] "Single-stranded Fv" is also abbreviated as "sFv" or "scFv," and refers to a single polypeptide chain to which V is attached. H antibody domain and V LIt is an antibody fragment containing an antibody domain. In some examples, the scFv polypeptide is further V H Domain and V L The scFv contains a binding polypeptide between the domain and the scFv, and the binding peptide allows the scFv to form an ideal antigen-binding structure. For an overview of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0064] The term "diabodies" is V H Domain and V L This is a small antibody fragment constructed by building an scFv fragment (see above) using short linkers (e.g., 5-10 residues) between domains. This causes the variable domains to pair between chains rather than within the chain, generating a bivalent fragment, i.e., a fragment with two antigen-binding sites. The bispecific diabody is a heterodimer of two "crossing" scFv fragments, and the V of two antibodies H Domain and V L The domains are located on different polypeptide chains. The diabodies are described in detail in EP 404,097;WO 93 / 11161;Hollinger et al., Proc.Natl.Acad.Sci.USA,90:6444-6448(1993).

[0065] The "humanized" form of a non-human (e.g., rodent) antibody is a chimeric antibody containing a minimal number of sequences derived from the non-human antibody. In most cases, the humanized antibody is a human immunoglobulin (receptor antibody), and the hypervariable region (HVR) residues of the receptor antibody are replaced with hypervariable region residues derived from a non-human species such as mouse, rat, rabbit, or non-human mammal, and which have ideal antibody specificity, affinity, and performance (donor antibody). In some cases, residues in the framework region of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, the humanized antibody may contain residues that are not present in either the receptor antibody or the donor antibody. These modifications can further improve the performance of the antibody. Generally, almost all humanized antibodies contain at least one, usually two, variable domains, all or almost all of their hypervariable loops correspond to the hypervariable loops of the non-human immunoglobulin, and all or almost all of their framework regions are human immunoglobulin sequences. Human antibodies optionally include at least a portion of the constant region (Fc) of an immunoglobulin, which is typically the constant region of human immunoglobulin. For further details, see Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).

[0066] The “percentage of amino acid sequence identity (%)” or “homology” of polypeptides and antibody sequences identified herein is defined as the percentage of identical amino acid residues in the polypeptide sequence being compared to the candidate sequence, when a sequence comparison is performed and the conservative substitution is considered to be part of the sequence identity. The percentage of amino acid sequence identity can be determined by several comparison methods in the art, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. Those skilled in the art can determine appropriate parameters for measurement and comparison, including any algorithm necessary to achieve a maximal comparison over the entire length of the sequences being compared. However, for the purposes of this specification, the numerical values ​​of the percentage of amino acid sequence identity were generated by the sequence comparison computer program MUSCLE (Edgar, RC, Nucleic Acids Research 32(5):1792-1797, 2004; Edgar, RC, BMC Bioinformatics 5(1):113, 2004).

[0067] The terms “Fc receptor” or “FcR” are used to describe receptors that bind to the Fc region of an antibody. In some examples, the FcRs described herein are FcRs that bind to an IgG antibody (one type of γ receptor) and include the FcγRI, FcγRII, and FcγRIII subclass receptors, including allelic variants and alternative splice forms of these receptors. The FcγRII receptors include FcγRIIA ("activating receptors") and FcγRIIB ("inhibiting receptors"), which have similar amino acid sequences but differ primarily in their cytoplasmic domains. The cytoplasmic domain of the activating receptor FcγRIIA contains an immunoreceptor tyrosine-activated motif (ITAM). The cytoplasmic domain of the inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-dependent inhibitory motif (ITIM) (see M. in Daeron Annu. Rev. Immunol. 15:203-234 (1997)). The above terms also include allotypes, for example, allotypes of FcγRIIIIA: FcγRIIIIA-Phe158, FcγRIIIIA-Val158, FcγRIIA-R131 and / or FcγRIIA-H131. For FcRs, see Ravetch and Kinet, Annu. Rev. Immunol. This is described in 9:457-92 (1991) and Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). In this specification, the term FcR includes other types of FcRs, including FcRs to be identified in the future. The term FcR also includes the neonatal receptor FcRn, which is responsible for the introduction of maternal IgGs into the neonatal body (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)).

[0068] The term "FcRn" refers to the neonatal Fc receptor (FcRn). FcRn is structurally similar to the major histocompatibility complex (MHC), with an α chain non-covalently bound to β2-microglobulin. The various functions of the neonatal Fc receptor FcRn are reviewed in Ghetie and Ward (2000) Annu. Rev. Immunol. 18, 739-766. FcRn plays an important role in the passive transport of immunoglobulin IgG from mother to neonatal and in the regulation of serum IgG levels. As a salvage receptor, FcRn can bind and transport endocytic IgG completely both intracellularly and intercellularly, protecting them from default degradation pathways.

[0069] The "CH1 domain" in the Fc region of human IgG generally extends from amino acid position 118 to amino acid position 215 (EU numbering system).

[0070] The "hinge region" is typically defined as extending from Glu at position 216 to Pro at position 230 in human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). By positioning the first and last cysteine ​​residues that form the inter-heavy chain disulfide bond in the same positions as in IgG1, the hinge regions of other IgG isotypes can be compared to the sequence of IgG1.

[0071] The "CH2 domain" in the Fc region of human IgG generally extends from amino acid position 231 to amino acid position 340. A unique feature of the CH2 domain is the insertion of two N-terminal branched sugar chains between the two CH2 domains in a complete, natural IgG molecule, without close pairing with other regions. It is hypothesized that these sugars may help maintain the stability of the CH2 domain as an alternative to pairing between domains. (Burton, Molec. Immunol. 22:161-206 (1985)).

[0072] The "CH3" domain includes a region extending from the C-terminal residue in the Fc region to the CH2 domain (from the amino acid at position 341 to the C-terminus of the antibody sequence, which is usually the amino acid residue at position 446 or 447 of IgG).

[0073] "Functional Fc fragments" possess the "effector functions" of native Fc region sequences. Exemplary "effector functions" include C1q binding, complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, and downregulation of cell surface receptors (e.g., B cell receptors, BCRs). Such effector functions typically require binding of the Fc region to a binding domain (e.g., an antibody variable region) and can be evaluated using various experimental methods known in the art.

[0074] Antibodies of IgG Fc variants with "modified" FcR binding affinity or ADCC activity exhibit enhanced or reduced FcR binding activity and / or ADCC activity compared to the parent polypeptide or polypeptide containing the natural Fc sequence. Fc variants showing "enhanced binding" to FcR exhibit higher binding affinity to at least one FcR (e.g., lower apparent Kd value or IC2) compared to the parent polypeptide or polypeptide containing the natural IgG Fc sequence. 50 It has a value. In some examples, the binding ability is increased by 3 times, for example, 5 times, 10 times, 25 times, 50 times, 60 times, 100 times, 150 times, 200 times, and even 500 times compared to the parent polypeptide, or the binding strength is increased by 25% to 1000%. Fc variants that show "reduced binding" to FcR have lower affinity (e.g., higher apparent Kd value or IC) to at least one FcR compared to the parent polypeptide. 50 It has a value of 40% or more compared to the parent polypeptide.

[0075] Antibody-dependent cell-mediated cytotoxicity, or ADCC, is a form of cytotoxicity in which secreted Ig antibodies bind to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer cells (NKs), neutrophils, and macrophages). These cytotoxic effector cells can then specifically bind to target cells containing antigens, subsequently killing the target cells using cytotoxins. Antibodies "arm" the cytotoxic cells, which is necessary for such killing. Of the main cell types that mediate ADCC, NK cells express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991), summarizes FcR expression on hematopoietic cells. To evaluate the ADCC activity of a target molecule, in vitro ADCC experiments can be performed, as described in U.S. Patent No. 5,500,362 or 5,821,337. Suitable effector cells for such experiments include peripheral blood mononuclear cells (PBMCs) and natural killer cells (NK). Optionally, or further, the ADCC activity of a target molecule can also be evaluated in vivo, as described, for example, in the animal model disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).

[0076] Polypeptides containing Fc region variants have been shown to exhibit "enhanced ADCC activity" or more effectively mediate ADCC effectors in the presence of human effector cells compared to polypeptides containing wild-type IgG Fc or the parent polypeptide. These Fc region variant polypeptides can more effectively mediate ADCC both in vivo and in vitro when their quantities are approximately the same as those of the wild-type IgG Fc polypeptide (or parent polypeptide) at the time of the experiment. Generally, any in vitro ADCC experimental method known in the art is used to identify such variants, such as experiments or methods for identifying ADCC activity, or, for example, animal models. In some examples, such variants show a 5-100-fold, or for example, 25-50-fold, increase in ADCC-mediating efficiency compared to wild-type Fc (or parent polypeptide).

[0077] "Complement-dependent cell injury" or "CDC" means lysing target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to an antibody (a subclass with the appropriate structure) that binds to a congener antigen. To evaluate complement activation, CDC experiments can be performed, as described, for example, in Gazzano-Santoro et al., J.Immunol.Methods 202:163 (1996). Polypeptide variants with modified Fc region amino acid sequences and increased or decreased C1q binding ability are described in U.S. Patent Nos. 6,194,551B1 and WO99 / 51642. The contents of these patent publications are explicitly incorporated herein by reference. See also Idusougie et al. J.Immunol.164:4178-4184 (2000).

[0078] Unless otherwise specified, one of the "nucleotide sequences encoding an amino acid sequence" includes all nucleotide sequences that are degenerate forms of each other and that encode the same amino acid sequence. Nucleotide sequences encoding proteins or RNA may contain introns; for example, nucleotide sequences encoding proteins contain introns in several forms.

[0079] The term "operably linked" means that a sequence is expressed by regulating the functional linkage between it and a heterologous nucleotide sequence. For example, if a first nucleotide sequence and a second nucleotide sequence have a functional relationship, then the first and second nucleotide sequences are operably linked. For example, if a promoter influences the transcription or expression of a coding sequence, then the promoter and the coding sequence are operably linked. Generally, operably linked DNA sequences are contiguous and, if necessary, can link two protein-coding regions into the same reading frame.

[0080] "Homologie" refers to the sequence similarity or identity between two polypeptides or two nucleic acid molecules. Two comparison sequences are homologous at the same position if those positions are the same base or amino acid monomer subunit. For example, if two DNA molecules both have adenine at the same position, these two DNA molecules are homologous at that position. The percentage of homology between two sequences is a function of the ratio of the number of matching or homologous positions common to both sequences to the total number of positions, multiplied by 100. For example, if two sequences have 6 matching or homologous positions out of 10, the homology between these two sequences is 60%. For example, the DNA sequences ATTGCC and TATGGC have 50% homology. Typically, when comparing two sequences, the comparison is made to obtain the maximum possible homology.

[0081] The “effective amount” of any anti-CD40 antibody or composition disclosed herein means an amount sufficient to achieve a particular purpose. The “effective amount” can be determined by empirical and known methods relating to the purpose.

[0082] The term "therapeutic dose" means an amount of the anti-CD40 antibody or composition disclosed herein that can effectively treat a disease or condition in an individual. That is, an amount sufficient to reduce or improve the severity and / or duration of a disease or one or more symptoms thereof, to prevent disease progression, to cause disease remission, to prevent recurrence, development, seizure, or progression of one or more symptoms associated with the disease, to detect the disease, or to enhance / improve the preventive or therapeutic effect of another treatment (e.g., a prophylactic or therapeutic agent).

[0083] As used herein, “pharmaceutically acceptable” or “pharmacologically compatible” means a material that is free from biological activity or other undesirable properties, such as being able to be added to a pharmaceutical composition administered to a patient without causing a significant adverse biological reaction or interacting in an adverse manner with any other components of the composition. A pharmaceutically acceptable carrier or excipient is preferably one that meets the standards required for toxicological or manufacturing testing and / or is included in the inactive ingredient guidelines created by the U.S. Food and Drug Administration.

[0084] The embodiments of the present invention described herein should be understood to include embodiments comprising "..." and / or "substantially comprising...".

[0085] As used herein, “about” refers to a number or parameter, and includes (and describes) variations of the number or parameter itself. For example, a description of “about X” includes a description of “X”.

[0086] A statement that a particular number or parameter is "not" usually means "other than" that number or parameter. For example, "this method cannot be used to treat type X cancer" means that this method is typically used to treat types of cancer other than type X cancer.

[0087] Unless explicitly stated in another paragraph, the singular forms “1,” “one,” and “the said” as used herein and in the claims include multiple subjects.

[0088] Anti-CD40 antibody According to one embodiment, the present invention provides an anti-CD40 antibody that specifically binds to CD40. The anti-CD40 antibody includes, but is not limited to, humanized antibodies, chimeric antibodies, mouse antibodies, human antibodies, and antibody molecules comprising heavy-chain and / or light-chain CDRs as described herein. According to one embodiment, the present invention provides an isolated antibody that binds to CD40. Expected anti-CD40 antibodies include, for example, full-length anti-CD40 antibodies (e.g., full-length IgG1 or full-length IgG4), anti-CD40 single-chain antibodies, anti-CD40Fc fusion proteins, multispecific (e.g., bispecific) anti-CD40 antibodies, and anti-CD40 immunoconjugates. In some examples, the anti-CD40 antibody is a full-length antibody (e.g., full-length IgG1 or full-length IgG4) or its antigen-binding fragment that specifically binds to CD40. In some examples, the anti-CD40 antibody is Fab, Fab', F(ab)'2, Fab'-SH, single-chain Fv(scFv), Fv fragment, dAb, Fd, nanobody, diabody, or linear antibody. In some examples, the antibody that specifically binds to CD40 has a binding affinity between the antibody and CD40 that is at least 10 times greater than the binding affinity between the antibody and the non-target (e.g., 10, 10). 2 , 10 3 , 10 4 , 10 5 , 10 6 , or 10 7This means that it includes multiples. In some examples, non-target means an antigen that is not CD40. Binding affinity can be measured by methods known in the art, such as ELISA, fluorescence-activated cell sorting (FACS) analysis, or radioimmunoassay (RIA). Kd value can be measured by methods known in the art, such as surface plasmon resonance (SPR) technique or biolayer interference (BLI) technique.

[0089] This specification has broadly discussed anti-CD40 antibodies containing human sequences (e.g., human heavy and light chain variable domains containing human CDR sequences), but has also considered non-human anti-CD40 antibodies. In some examples, the non-human anti-CD40 antibody comprises the human CDR sequence and non-human framework region sequence of the anti-CD40 antibody described herein, and in some examples, the non-human framework region sequence comprises any sequence for generating heavy and / or light chain variable domains using one or more human CDR sequences described herein, including mammals such as mice, rats, rabbits, pigs, cattle (e.g., cattle, yaks, buffalo), deer, sheep, goats, chickens, cats, dogs, ferrets, and primates (e.g., marmosets, macaques). In some examples, the non-human anti-CD40 antibody comprises an anti-CD40 antibody produced by transplanting one or more human CDR sequences described herein into a non-human framework region (e.g., a mouse or chicken framework region sequence).

[0090] The complete amino acid sequence of exemplary human CD40 includes or consists of the amino acid sequence shown in SEQ ID NO:149.

[0091] In some examples, the anti-CD40 antibodies described herein specifically recognize epitopes in human CD40. In some examples, the anti-CD40 antibodies cross-react with CD40 from other non-human species. In some examples, the anti-CD40 antibodies are completely specific to human CD40 and do not cross-react with other non-human species.

[0092] In some examples, the anti-CD40 antibody cross-reacts with at least one allele variant of the CD40 protein (or fragment thereof). In some examples, the allele variant has at most 30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or 30) amino acid substitutions (e.g., conserved substitutions) compared to the naturally occurring CD40 protein (or fragment thereof). In some examples, the anti-CD40 antibody does not cross-react with any allele variant of the CD40 protein (or fragment thereof).

[0093] In some examples, the anti-CD40 antibody cross-reacts with at least one interspecies variant of the CD40 protein. In some examples, for instance, the CD40 protein (or its fragment) is human CD40, and the interspecies variant of the CD40 protein (or its fragment) is a variant in cynomolgus monkeys. In some examples, the anti-CD40 antibody does not cross-react with any interspecies variant of the CD40 protein.

[0094] In some examples, any anti-CD40 antibody described herein comprises an antibody heavy chain constant region and an antibody light chain constant region. In some examples, the anti-CD40 antibody comprises an IgG1 type heavy chain constant region. In some examples, the anti-CD40 antibody comprises an IgG2 type heavy chain constant region. In some examples, the anti-CD40 antibody comprises an IgG3 type heavy chain constant region. In some examples, the anti-CD40 antibody comprises an IgG4 type heavy chain constant region. In some examples, the heavy chain constant region comprises amino acid sequence SEQ ID NO: 145 (including consisting of... or substantially consisting of...). In some examples, the heavy chain constant region comprises amino acid sequence SEQ ID NO: 146 (including consisting of... or substantially consisting of...). In some examples, the anti-CD40 antibody comprises a κ light chain constant region. In some examples, the light chain constant region comprises amino acid sequence SEQ ID NO: 147 (including consisting of... or substantially consisting of...). In some examples, the anti-CD40 antibody includes a λ light chain constant region. In some examples, the light chain constant region includes the amino acid sequence SEQ ID NO:148 (or consists of..., or substantially consists of...). In some examples, the anti-CD40 antibody includes an antibody heavy chain variable domain and an antibody light chain variable domain.

[0095] In some examples, the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V HThis includes a heavy chain complementarity determination region (HC-CDR) 1, HC-CDR 2, and HC-CDR 3, wherein the heavy chain complementarity determination region (HC-CDR) 1 includes GYX1VN (SEQ ID NO: 81), where X1 is L, T, or V; the HC-CDR 2 includes X1IX2PX3X4GX5TX6YNQKFKG (SEQ ID NO: 82), where X1 is L or M, X2 is A, N, or V, X3 is E, K, L, M, R, S, T, or Y, X4 is H, N, S, or T, X5 is G or R, X6 is A, S, T, or W; and the HC-CDR 3 includes SX1X2X3X4PYAX5DY (SEQ ID NO: 82). NO:83) includes, where X1 is G, I, L, M, P, R, S, or Y, X2 is W, or Y, X3 is S, or V, X4 is D, K, L, N, R, T, or V, and X5 is A, G, M, N, Q, or S, and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 is KASQNV H TX1VA (SEQ ID NO: 84) is included, where X1 is A, H, N, or R; LC-CDR2 is included, where LX1SDRRT (SEQ ID NO: 85) is included, where X1 is A or R; LC-CDR3 is included, where LQX1WX2X3PLT (SEQ ID NO: 86) is included, where X1 is A or H, X2 is N, S, or T, and X3 is A, Q, R, S, or Y.

[0096] In some examples, the anti-CD40 antibody was V H Including the above V HThe present invention comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs:1-3 or a variant of the amino acid sequence shown in any of SEQ ID NOs:1-3, comprising at most about three (e.g., one, two, or three) amino acid substitutions; HC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs:9-26 or a variant of the amino acid sequence shown in any of SEQ ID NOs:9-26, comprising at most about three (e.g., one, two, or three) amino acid substitutions; and HC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs:32-52 or a variant of the amino acid sequence shown in any of SEQ ID NOs:32-52, comprising at most about three (e.g., one, two, or three) amino acid substitutions.

[0097] In some examples, the anti-CD40 antibody was V H Including the above V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs: 1-3, HC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs: 9-26, and HC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs: 32-52.

[0098] In some examples, the anti-CD40 antibody was V L Including the above V LThe present invention comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs: 58-61 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-61, comprising at most about three (e.g., one, two, or three) amino acid substitutions; LC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs: 65-66 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 65-66, comprising at most about three (e.g., one, two, or three) amino acid substitutions; and LC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs: 70-77 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 70-77, comprising at most about three (e.g., one, two, or three) amino acid substitutions.

[0099] In some examples, the anti-CD40 antibody was V L Including the above V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs: 58-61, LC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs: 65-66, and LC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs: 70-77.

[0100] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs:1-3 or a variant of the amino acid sequence shown in any of SEQ ID NOs:1-3, comprising at most about three (e.g., one, two, or three) amino acid substitutions; HC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs:9-26 or a variant of the amino acid sequence shown in any of SEQ ID NOs:9-26, comprising at most about three (e.g., one, two, or three) amino acid substitutions; and HC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs:32-52 or a variant of the amino acid sequence shown in any of SEQ ID NOs:32-52, comprising at most about three (e.g., one, two, or three) amino acid substitutions. L The present invention comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs: 58-61 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-61, comprising at most about three (e.g., one, two, or three) amino acid substitutions; LC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs: 65-66 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 65-66, comprising at most about three (e.g., one, two, or three) amino acid substitutions; and LC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs: 70-77 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 70-77, comprising at most about three (e.g., one, two, or three) amino acid substitutions.

[0101] In some examples, the anti-CD40 antibody was V H and V L Including the above V HIt comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs: 1-3, HC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs: 9-26, and HC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs: 32-52, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises an amino acid sequence shown in any of SEQ ID NOs: 58-61, LC-CDR2 comprises an amino acid sequence shown in any of SEQ ID NOs: 65-66, and LC-CDR3 comprises an amino acid sequence shown in any of SEQ ID NOs: 70-77.

[0102] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:9, and HC-CDR3 comprises amino acid sequence SEQ ID NO:32, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 58, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0103] In some examples, the anti-CD40 antibody was V H and V L Including the above VH This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:9, and HC-CDR3 comprises amino acid sequence SEQ ID NO:32, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 58, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70.

[0104] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:10, and HC-CDR3 comprises amino acid sequence SEQ ID NO:32. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0105] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:10, and HC-CDR3 comprises amino acid sequence SEQ ID NO:32. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70.

[0106] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:13, and HC-CDR3 comprises amino acid sequence SEQ ID NO:34. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0107] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:13, and HC-CDR3 comprises amino acid sequence SEQ ID NO:34. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70.

[0108] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:37, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L This includes LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 73. L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0109] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:37, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 contains amino acid sequence SEQ ID NO: 59, LC-CDR2 contains amino acid sequence SEQ ID NO: 65, and LC-CDR3 contains amino acid sequence SEQ ID NO: 73.

[0110] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:20, and HC-CDR3 comprises amino acid sequence SEQ ID NO:42, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0111] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:20, and HC-CDR3 comprises amino acid sequence SEQ ID NO:42, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70.

[0112] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:14, and HC-CDR3 comprises amino acid sequence SEQ ID NO:43. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0113] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:14, and HC-CDR3 comprises amino acid sequence SEQ ID NO:43. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70.

[0114] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:51. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 70, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0115] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:51. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 70.

[0116] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:11, and HC-CDR3 comprises amino acid sequence SEQ ID NO:48. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 71, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0117] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:11, and HC-CDR3 comprises amino acid sequence SEQ ID NO:48. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 71.

[0118] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:18, and HC-CDR3 comprises amino acid sequence SEQ ID NO:47, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 59, LC-CDR2 comprises amino acid sequence SEQ ID NO: 65, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 71, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0119] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:18, and HC-CDR3 comprises amino acid sequence SEQ ID NO:47, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 59, LC-CDR2 includes amino acid sequence SEQ ID NO: 65, and LC-CDR3 includes amino acid sequence SEQ ID NO: 71.

[0120] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes the amino acid sequence shown in any of SEQ ID NOs: 1-3, 9-26, and 32-52, or a variant of the amino acid sequence shown in any of SEQ ID NOs: 1-3, 9-26, and 32-52, which includes at most about five amino acid substitutions, and the V L This includes the amino acid sequence shown in any of SEQ ID NOs: 58-61, 65-66, and 70-77, or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-61, 65-66, and 70-77, which includes at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes an amino acid sequence shown in any of SEQ ID NOs: 1-3, 9-26, and 32-52, and the V L This includes the amino acid sequence shown in SEQ ID NOs: 58-61, 65-66, and 70-77.

[0121] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the VH This includes amino acid sequences SEQ ID NOs: 1, 9, and 32, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 58, 65, and 70, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 9, and 32, and the V L This includes amino acid sequences SEQ ID NOs: 58, 65, and 70.

[0122] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 1, 10, and 32, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 10, and 32, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70.

[0123] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V HThis includes amino acid sequences SEQ ID NOs: 1, 13, and 34, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 13, and 34, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70.

[0124] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 1, 15, and 37, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 73, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 15, and 37, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 73.

[0125] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V HThis includes amino acid sequences SEQ ID NOs: 1, 20, and 42, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 20, and 42, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70.

[0126] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 1, 14, and 43, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 14, and 43, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70.

[0127] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V HThis includes amino acid sequences SEQ ID NOs: 1, 15, and 51, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 15, and 51, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 70.

[0128] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 1, 11, and 48, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 71, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 11, and 48, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 71.

[0129] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V HThis includes amino acid sequences SEQ ID NOs: 1, 18, and 47, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 71, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 1, 18, and 47, and the V L This includes amino acid sequences SEQ ID NOs: 59, 65, and 71.

[0130] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This refers to V, which is represented by one of the amino acid sequences of SEQ ID NOs: 87-119. H The V L This refers to V, which is represented by one of the amino acid sequences of SEQ ID NOs:126-138. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0131] In some examples, the anti-CD40 antibody was V H Including the above V H This includes one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:87. In some examples, the anti-CD40 antibody is V H Including the above V H This includes one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:88. In some examples, the anti-CD40 antibody is V H Including the above V HThis includes one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:89. In some examples, the anti-CD40 antibody is V H Including the above V H This includes one, two, or three HC-CDRs in the amino acid sequence SEQ ID NO:94. In some examples, the anti-CD40 antibody is V H Including the above V H This includes one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:96. In some examples, the anti-CD40 antibody is V H Including the above V H This includes one, two, or three HC-CDRs in the amino acid sequence SEQ ID NO:101. In some examples, the anti-CD40 antibody is V H Including the above V H This includes one, two, or three HC-CDRs in the amino acid sequence SEQ ID NO:110. In some examples, the anti-CD40 antibody is V H Including the above V H This includes one, two, or three HC-CDRs in the amino acid sequence SEQ ID NO:114. In some examples, the anti-CD40 antibody is V H Including the above V H This includes one, two, or three HC-CDRs in the amino acid sequence SEQ ID NO:115. In some examples, the anti-CD40 antibody is V H Including the above V H This contains one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:117.

[0132] In some examples, the anti-CD40 antibody was V L Including the above V L This includes one, two, or three LC-CDRs in amino acid sequence SEQ ID NO:126. In some examples, the anti-CD40 antibody is V L Including the above V LThis includes one, two, or three LC-CDRs in the amino acid sequence SEQ ID NO:127. In some examples, the anti-CD40 antibody is V L Including the above V L This includes one, two, or three LC-CDRs in amino acid sequence SEQ ID NO:128. In some examples, the anti-CD40 antibody is V L Including the above V L This includes one, two, or three LC-CDRs in the amino acid sequence SEQ ID NO:129. In some examples, the anti-CD40 antibody is V L Including the above V L This includes one, two, or three LC-CDRs in amino acid sequence SEQ ID NO:132.

[0133] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:87. H The V L This is the V shown in amino acid sequence SEQ ID NO:126. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0134] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:88. H The V L This is the V shown in amino acid sequence SEQ ID NO:127. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0135] In some examples, the anti-CD40 antibody was VH and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:89. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0136] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:94. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0137] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:96. H The V L This is the V shown in amino acid sequence SEQ ID NO:132. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0138] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:101. H The V LThis is the V shown in amino acid sequence SEQ ID NO:128. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0139] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:110. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0140] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:114. H The V L This is the V shown in amino acid sequence SEQ ID NO:128. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0141] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:115. H The V L This is the V shown in amino acid sequence SEQ ID NO:129. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0142] In some examples, the anti-CD40 antibody was V H and VL Including the above V H This is the V shown in amino acid sequence SEQ ID NO:117. H The V L This is the V shown in amino acid sequence SEQ ID NO:129. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0143] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes an amino acid sequence shown in any of SEQ ID NOs:87-119 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with an amino acid sequence shown in any of SEQ ID NOs:87-119, and the V L This includes the amino acid sequence shown in any of SEQ ID NOs:126-138 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence shown in any of SEQ ID NOs:126-138. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence shown in any of SEQ ID NOs:87-119, and the V L It contains the amino acid sequence shown in one of the SEQ ID NOs:126-138.

[0144] In some examples, the anti-CD40 antibody was V H and V L Including the above V HThis includes amino acid sequence SEQ ID NO:87 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:87, and the V L This includes amino acid sequence SEQ ID NO:126 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:126. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:87, and the V L This includes amino acid sequence SEQ ID NO:126.

[0145] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:88 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:88, and the V L This includes amino acid sequence SEQ ID NO:127 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:127. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:88, and the V L This contains amino acid sequence SEQ ID NO:127.

[0146] In some examples, the anti-CD40 antibody was V H and V L Including the above VH This includes amino acid sequence SEQ ID NO:89 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:89, and the V L This includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:89, and the V L This includes amino acid sequence SEQ ID NO:128.

[0147] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:94 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:94, and the V L This includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:94, and the V L This includes amino acid sequence SEQ ID NO:128.

[0148] In some examples, the anti-CD40 antibody was V H and V LIncluding the above V H This includes amino acid sequence SEQ ID NO:96 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:96, and the V L This includes amino acid sequence SEQ ID NO:132 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:132. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:96, and the V L This contains amino acid sequence SEQ ID NO:132.

[0149] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:101 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:101, and the V L This includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:101, and the V L This includes amino acid sequence SEQ ID NO:128.

[0150] In some examples, the anti-CD40 antibody was V Hand V L Including the above V H This includes amino acid sequence SEQ ID NO:110 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:110, and the V L This includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:110, and the V L This includes amino acid sequence SEQ ID NO:128.

[0151] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:114 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:114, and the V L This includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:114, and the V L This includes amino acid sequence SEQ ID NO:128.

[0152] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:115 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:115, and the V L This includes amino acid sequence SEQ ID NO:129 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:129. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:115, and the V L This includes amino acid sequence SEQ ID NO:129.

[0153] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:117 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:117, and the V L This includes amino acid sequence SEQ ID NO:129 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:129. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:117, and the V L This includes amino acid sequence SEQ ID NO:129.

[0154] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:4, HC-CDR2 comprises amino acid sequence SEQ ID NO:27, and HC-CDR3 comprises amino acid sequence SEQ ID NO:53. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:67, and LC-CDR3 comprises amino acid sequence SEQ ID NO:78. L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0155] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:4, HC-CDR2 comprises amino acid sequence SEQ ID NO:27, and HC-CDR3 comprises amino acid sequence SEQ ID NO:53. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO:62, LC-CDR2 includes amino acid sequence SEQ ID NO:67, and LC-CDR3 includes amino acid sequence SEQ ID NO:78.

[0156] In some examples, the anti-CD40 antibody was V H or VH mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 4, 27, and 53, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 62, 67, and 78, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 4, 27, and 53, and the V L This includes amino acid sequences SEQ ID NOs: 62, 67, and 78.

[0157] In some examples, the anti-CD40 antibody was V H Including the above V H This contains one, two, or three HC-CDRs in the amino acid sequence SEQ ID NO:120.

[0158] In some examples, the anti-CD40 antibody was V L Including the above V L This includes one, two, or three LC-CDRs in amino acid sequence SEQ ID NO:139.

[0159] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:120. H The V L This is the V shown in amino acid sequence SEQ ID NO:139. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0160] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:120 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:120, and the V L This includes amino acid sequence SEQ ID NO:139 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:139. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:120, and the V L This includes amino acid sequence SEQ ID NO:139.

[0161] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO: 5, HC-CDR2 comprises amino acid sequence SEQ ID NO: 27, and HC-CDR3 comprises amino acid sequence SEQ ID NO: 54. H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:67, and LC-CDR3 comprises amino acid sequence SEQ ID NO:78. LThe LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0162] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO: 5, HC-CDR2 comprises amino acid sequence SEQ ID NO: 27, and HC-CDR3 comprises amino acid sequence SEQ ID NO: 54. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO:62, LC-CDR2 includes amino acid sequence SEQ ID NO:67, and LC-CDR3 includes amino acid sequence SEQ ID NO:78.

[0163] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 5, 27, and 54, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 62, 67, and 78, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 5, 27, and 54, and the V L This includes amino acid sequences SEQ ID NOs: 62, 67, and 78.

[0164] In some examples, the anti-CD40 antibody was V H Including the above VH This contains one, two, or three HC-CDRs in the amino acid sequence SEQ ID NO:121.

[0165] In some examples, the anti-CD40 antibody was V L Including the above V L This includes one, two, or three LC-CDRs in amino acid sequence SEQ ID NO:140.

[0166] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:121. H The V L This is the V shown in amino acid sequence SEQ ID NO:140. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0167] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:121 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:121, and the V L This includes amino acid sequence SEQ ID NO:140 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:140. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:121, and the V L This contains amino acid sequence SEQ ID NO:140.

[0168] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:6, HC-CDR2 comprises amino acid sequence SEQ ID NO:28, and HC-CDR3 comprises amino acid sequence SEQ ID NO:55, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:63, LC-CDR2 comprises amino acid sequence SEQ ID NO:68, and LC-CDR3 comprises amino acid sequence SEQ ID NO:79, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0169] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:6, HC-CDR2 comprises amino acid sequence SEQ ID NO:28, and HC-CDR3 comprises amino acid sequence SEQ ID NO:55, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 63, LC-CDR2 includes amino acid sequence SEQ ID NO: 68, and LC-CDR3 includes amino acid sequence SEQ ID NO: 79.

[0170] In some examples, the anti-CD40 antibody was V Hor V H mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 6, 28, and 55, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 63, 68, and 79, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 6, 28, and 55, and the V L This includes amino acid sequences SEQ ID NOs: 63, 68, and 79.

[0171] In some examples, the anti-CD40 antibody was V H Including the above V H This includes one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:122.

[0172] In some examples, the anti-CD40 antibody was V L Including the above V L This includes one, two, or three LC-CDRs in the amino acid sequence SEQ ID NO:141.

[0173] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:122. H The V L This is the V shown in amino acid sequence SEQ ID NO:141. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0174] In some embodiments, the anti-CD40 antibody is V H and V L and the V H comprises the amino acid sequence SEQ ID NO:122 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence SEQ ID NO:122, and the V L comprises the amino acid sequence SEQ ID NO:141 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence SEQ ID NO:141. In some embodiments, the anti-CD40 antibody is V H and V L and the V H comprises the amino acid sequence SEQ ID NO:122, and the V L comprises the amino acid sequence SEQ ID NO:141.

[0175] In some embodiments, the anti-CD40 antibody comprises a variant of V H or the V H and a variant of V L or the V L and the V H comprises HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprises the amino acid sequence SEQ ID NO:7, the HC-CDR2 comprises the amino acid sequence SEQ ID NO:29, the HC-CDR3 comprises the amino acid sequence SEQ ID NO:56, the HC-CDRs of the variant of the V H comprise at most about 5 amino acid substitutions, and the V L comprises LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprises the amino acid sequence SEQ ID NO:64, the LC-CDR2 comprises the amino acid sequence SEQ ID NO:69, the LC-CDR3 comprises the amino acid sequence SEQ ID NO:80, and the V LThe LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0176] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:7, HC-CDR2 comprises amino acid sequence SEQ ID NO:29, and HC-CDR3 comprises amino acid sequence SEQ ID NO:56. L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 64, LC-CDR2 includes amino acid sequence SEQ ID NO: 69, and LC-CDR3 includes amino acid sequence SEQ ID NO: 80.

[0177] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 7, 29, and 56, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 64, 69, and 80, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 7, 29, and 56, and the V L This includes amino acid sequences SEQ ID NOs: 64, 69, and 80.

[0178] In some examples, the anti-CD40 antibody was V H Including the above VH This contains one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:123.

[0179] In some examples, the anti-CD40 antibody was V L Including the above V L This includes one, two, or three LC-CDRs in amino acid sequence SEQ ID NO:142.

[0180] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:123. H The V L This is the V shown in amino acid sequence SEQ ID NO:142. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0181] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:123 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:123, and the V L This includes amino acid sequence SEQ ID NO:142 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:142. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:123, and the V L This includes amino acid sequence SEQ ID NO:142.

[0182] In some examples, the anti-CD40 antibody was V H or V H mutants of V L or V L Includes variants of the V H This includes HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 includes amino acid sequence SEQ ID NO:8, HC-CDR2 includes amino acid sequence SEQ ID NO:30, and HC-CDR3 includes amino acid sequence SEQ ID NO:57, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:67, and LC-CDR3 comprises amino acid sequence SEQ ID NO:80, and V L The LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0183] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 includes amino acid sequence SEQ ID NO:8, HC-CDR2 includes amino acid sequence SEQ ID NO:30, and HC-CDR3 includes amino acid sequence SEQ ID NO:57, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO:62, LC-CDR2 includes amino acid sequence SEQ ID NO:67, and LC-CDR3 includes amino acid sequence SEQ ID NO:80.

[0184] In some examples, the anti-CD40 antibody was V Hand V L comprises, and said V H comprises the amino acid sequences SEQ ID NOs: 8, 30, and 57, or said V H comprises variants thereof, said variants comprising at most about 5 amino acid substitutions, and said V L comprises the amino acid sequences SEQ ID NOs: 62, 67, and 80, or said V L comprises variants thereof, said variants comprising at most about 5 amino acid substitutions. In some embodiments, said anti-CD40 antibody comprises V H and V L comprises, and said V H comprises the amino acid sequences SEQ ID NOs: 8, 30, and 57, and said V L comprises the amino acid sequences SEQ ID NOs: 62, 67, and 80.

[0185] In some embodiments, said anti-CD40 antibody comprises V H comprises, and said V H comprises one, two, or three HC-CDRs in amino acid sequence SEQ ID NO: 124.

[0186] In some embodiments, said anti-CD40 antibody comprises V L comprises, and said V L comprises one, two, or three LC-CDRs in amino acid sequence SEQ ID NO: 143.

[0187] In some embodiments, said anti-CD40 antibody comprises V H and V L comprises, and said V H comprises HC-CDR1, HC-CDR2, and HC-CDR3 contained in V shown in amino acid sequence SEQ ID NO: 124, and said V H comprises LC-CDR1, LC-CDR2, and LC-CDR3 contained in V shown in amino acid sequence SEQ ID NO: 143. L comprises, and said V L

[0188] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:124 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:124, and the V L This includes amino acid sequence SEQ ID NO:143 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:143. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:124, and the V L This contains amino acid sequence SEQ ID NO:143.

[0189] In some examples, the anti-CD40 antibody was V H or the aforementioned V H mutants of V L or the aforementioned V L Includes variants of the V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:4, HC-CDR2 comprises amino acid sequence SEQ ID NO:31, and HC-CDR3 comprises amino acid sequence SEQ ID NO:53, and V H The HC-CDRs of the mutants contain at most about 5 amino acid substitutions, and the V L It comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:68, and LC-CDR3 comprises amino acid sequence SEQ ID NO:79, and V LThe LC-CDRs of the mutants contain at most about five amino acid substitutions.

[0190] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This comprises HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:4, HC-CDR2 comprises amino acid sequence SEQ ID NO:31, and HC-CDR3 comprises amino acid sequence SEQ ID NO:53, and V L This comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 includes amino acid sequence SEQ ID NO: 62, LC-CDR2 includes amino acid sequence SEQ ID NO: 68, and LC-CDR3 includes amino acid sequence SEQ ID NO: 79.

[0191] In some examples, the anti-CD40 antibody was V H or the aforementioned V H mutants of V L or the aforementioned V L Includes variants of the V H This includes amino acid sequences SEQ ID NOs: 4, 31, and 53, and the V H The variants contain at most about five amino acid substitutions, and the V L This includes amino acid sequences SEQ ID NOs: 62, 68, and 79, and the V L The variants contain at most about five amino acid substitutions. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequences SEQ ID NOs: 4, 31, and 53, and the V L This includes amino acid sequences SEQ ID NOs: 62, 68, and 79.

[0192] In some examples, the anti-CD40 antibody was V H Including the above VH This includes one, two, or three HC-CDRs in amino acid sequence SEQ ID NO:125.

[0193] In some examples, the anti-CD40 antibody was V L Including the above V L This includes one, two, or three LC-CDRs in amino acid sequence SEQ ID NO:144.

[0194] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This is the V shown in amino acid sequence SEQ ID NO:125. H The V L This is the V shown in amino acid sequence SEQ ID NO:144. L This includes LC-CDR1, LC-CDR2, and LC-CDR3.

[0195] In some examples, the anti-CD40 antibody was V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:125 or a mutant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:125, and the V L This includes amino acid sequence SEQ ID NO:144 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with amino acid sequence SEQ ID NO:144. In some examples, the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:125, and the V L This includes amino acid sequence SEQ ID NO:144.

[0196] In some examples, the amino acid substitutions are limited to the “exemplary substitutions” shown in Table 4 of this specification. In some examples, the amino acid substitutions are limited to the “preferred substitutions” shown in Table 4 of this specification.

[0197] In some examples, functional epitopes can be analyzed by complex alanine scanning. In this process, complex alanine scanning technology is used to identify the amino acids in the CD40 protein that are necessary for interaction with anti-CD40 antibodies. In some examples, the epitope is a three-dimensional structure, and simultaneously, the epitope can be identified using the crystal structure of the anti-CD40 antibody bound to the CD40 protein.

[0198] In some examples, the present invention provides antibodies that bind to CD40 competitively with any of the anti-CD40 antibodies described herein. In some examples, the present invention provides antibodies that can bind to epitopes on CD40 competitively with any of the anti-CD40 antibodies described herein. In some examples, V H and V L The present invention provides an anti-CD40 antibody that binds to the same epitope as the anti-CD40 antibody molecule containing the V H This includes the amino acid sequence shown in any of SEQ ID NOs:87-125, and the V L This includes the amino acid sequence shown in any of SEQ ID NOs:126-144. In some examples, V H and V L The present invention provides an anti-CD40 antibody that competitively binds to CD40 with anti-CD40 antibodies containing the above V H This includes the amino acid sequence shown in any of SEQ ID NOs:87-125, and the V L It contains the amino acid sequence shown in one of the SEQ ID NOs:126-144.

[0199] In some examples, competitive testing can identify monoclonal antibodies that competitively bind to CD40 with the anti-CD40 antibodies described herein. Competitive testing can determine whether two antibodies bind to the same epitope by recognizing the same or spatially overlapping epitopes, or by one antibody competitively inhibiting the binding of another antibody to the antigen. In some examples, this competitive antibody binds to the same epitope as the antibody described herein. Some exemplary competitive testing includes, but is not limited to, the conventional testing described in Harlow and Lane (1988) Antibodies: A Laboratory Manual ch.14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY). Detailed exemplary methods for analyzing the epitopes to which antibodies bind are described in Morris (1996) "Epitope Mapping Protocols," in Methods in Molecular Biology vol.66 (Humana Press, Totowa, NJ). In some examples, if each antibody blocks more than 50% of the binding of the other antibody, it is said to bind to the same epitope. In some examples, antibodies that compete with the anti-CD40 antibodies described herein are chimeric antibodies, humanized antibodies, or fully human antibodies.

[0200] Exemplary anti-CD40 antibody sequences are shown in Tables 2 and 3, and assigned CDR numbers according to the Kabat definition scheme. Those skilled in the art will recognize that there are several known algorithms (Kabat definition schemes) for predicting the location of CDRs and determining the light and heavy chain variable regions of the antibody. The CDRs of the antibodies described herein are V H , and / or V L While including sequences, antibodies other than those exemplified in the table below are also within the scope of this invention, based on the prediction algorithm.

[0201] [Table 2] JPEG0007833531000003.jpg253170JPEG0007833531000004.jpg255168JPEG0007833531000005.jpg101170

[0202] Table 3 JPEG0007833531000007.jpg242170JPEG0007833531000008.jpg250170JPEG0007833531000009.jpg255170JPEG0007833531000010.jpg67170

[0203] CD40 CD40 is a 45-50 kD type I membrane protein and a member of the TNFR superfamily. CD40 is expressed not only in antigen-presenting cells (e.g., B cells, dendritic cells (DCs), macrophages, and monocytes) but also in non-immune cells such as epithelial, endothelial, and stromal cells (fibroblasts, myofibroblasts, synovial cells, astrocytes, etc.), platelets, and tumor cells (Schonbeck, U, and P Libby. Cellular and molecular life sciences: CMLS vol. 58, 1(2001): 4-43). As a transmembrane signaling molecule, the CD40 molecule triggers the activation of intracellular kinases and transcription factors within cells, and is involved in regulating a wide range of molecular and cellular processes, including the initiation and development of cellular and humoral adaptive immunity. Due to the lack of inherent kinase activity in the intracellular domain of CD40, its signaling is primarily transmitted by ligand-dependent recruitment of adapter proteins of the TNF receptor-related factor (TRAF) family. TRAF not only couples CD40 to intracellular signaling components, but also detaches this signaling complex from CD40, releases it into the cytoplasm, and activates proximal protein kinases as E3 ubiquitin ligases. TRAF can activate extracellular signal-regulated protein kinases (ERKs), c-Jun amino-terminal kinases (JNKs), and p38 MAPK, the phosphatidylinositol 3-kinase (PI3K) cascade, the transcription factor nuclear factor-κB (NF-κB), and mitogen-activated protein kinase (MAPK) pathways, including signaling and transcriptional activators (STATs) (Loskog, Angelica SI, and Aristides G Eliopoulos. Seminars in immunology vol.21,5(2009):301-7; and Matsuzawa, Atsushi et al. Science (New York, NY) vol.321,5889(2008):663-8). These pathways act synergistically to regulate CD40 activity in cell type and microenvironment-dependent forms.

[0204] The interaction between CD40 and CD40L triggers multiple immune responses, including the expression of cellular stress-related genes and B cell activation (immunoglobulin class switching, germinal center formation, and cytokine production) (Belkhir, R et al. Scandinavian journal of immunology vol.79,1(2014):37-42; and Bensinger, William et al. British journal of hematology vol.159,1(2012):58-66). The CD40-CD40L interaction also plays a crucial role in T cell activation and the upregulation of the expression of inflammation-related surface molecules and cytokines in chronic inflammatory diseases. These cytokines include interleukin (IL)-2, IL-12, TNF-α, interferon (IFN)-α, as well as IL-1 and IL-6 (Sidiropoulos, PI, and DT Boumpas. Lupus vol.13,5(2004):391-7).

[0205] Given the crucial role of CD40-CD40L as a co-stimulatory molecule between B cells and T cells, there is interest in the role of CD40-CD40L antagonists in inflammatory or autoimmune diseases (e.g., psoriasis, lupus nephritis, systemic lupus erythematosus (SLE), as well as rheumatoid arthritis (RA), diabetes, and immune rejection).

[0206] CD40L CD40L is a 32-39 kDa type II transmembrane protein and is also a member of the TNF ligand superfamily (TNFSF, which includes TNF, lymphotoxins, CD27L, CD30L, and FasL). CD40L is activated by CD4 +CD40L is expressed by T cells, as well as activated B cells and platelets, but under inflammatory conditions, variable expression can also be induced in monocytes, natural killer cells, mast cells, and basophils (Elgueta, Raul et al. Immunological reviews vol.229, 1(2009):152-72). Soluble CD40L (sCD40L) exhibits similar biological activity to its transmembrane form and has been reported to trigger the CD40L / CD40 signaling cascade.

[0207] Therefore, neutralizing the binding of CD40 to CD40L may be a potential therapeutic approach for diseases and symptoms mediated by the CD40-CD40L pathway.

[0208] Full-length anti-CD40 antibody In some embodiments, the anti-CD40 antibody is a full-length anti-CD40 antibody. In some embodiments, the full-length anti-CD40 antibody is IgA, IgD, IgE, IgG, or IgM. In some embodiments, the full-length anti-CD40 antibody includes an IgG constant region, which is, for example, IgG1, IgG2, IgG3, IgG4, or a variant thereof. In some embodiments, the full-length anti-CD40 antibody includes a λ light chain constant region. In some embodiments, the full-length anti-CD40 antibody includes a κ light chain constant region. In some embodiments, the full-length anti-CD40 antibody is a full-length human anti-CD40 antibody. In some embodiments, the full-length anti-CD40 antibody includes a mouse immunoglobulin Fc sequence. In some embodiments, the full-length anti-CD40 antibody has enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) effector function by including a modified or otherwise modified Fc sequence.

[0209] Therefore, for example, in some examples, a full-length anti-CD40 antibody containing an IgG1 constant region is provided, and the anti-CD40 antibody specifically binds to CD40. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0210] In some examples, a full-length anti-CD40 antibody containing an IgG2 constant region is provided, and the anti-CD40 antibody specifically binds to CD40. In some examples, the IgG2 is human IgG2. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0211] In some examples, a full-length anti-CD40 antibody containing an IgG3 constant region is provided, and the anti-CD40 antibody specifically binds to CD40. In some examples, the IgG3 is human IgG3. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0212] In some examples, a full-length anti-CD40 antibody containing an IgG4 constant region is provided, and the anti-CD40 antibody specifically binds to CD40. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region contains or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region contains or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region contains or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region contains or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region contains or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0213] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 1-8 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 1-8 comprising at most about three (e.g., one, two, or three) amino acid substitutions, the HC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 9-31 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 9-31 comprising at most about three (e.g., one, two, or three) amino acid substitutions, and the HC-CDR3 comprising SEQ ID The light chain variable domain comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises the amino acid sequence shown in any of SEQ ID NOs: 58-64 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-64, which includes at most about three (e.g., one, two, or three) amino acid substitutions, LC-CDR2 comprises the amino acid sequence shown in any of SEQ ID NOs: 65-69 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 65-69, which includes at most about three (e.g., one, two, or three) amino acid substitutions, and LC-CDR3 comprises SEQ ID The amino acid sequences include those represented by NOs:70-80 or variants of those amino acid sequences, including at most about three (e.g., one, two, or three) amino acid substitutions. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145.In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0214] In several examples, a full-length anti-CD40 antibody containing an IgG2 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 1-8 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 1-8 comprising at most about three (e.g., one, two, or three) amino acid substitutions, the HC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 9-31 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 9-31 comprising at most about three (e.g., one, two, or three) amino acid substitutions, and the HC-CDR3 comprising SEQ ID The light chain variable domain comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises the amino acid sequence shown in any of SEQ ID NOs: 58-64 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-64, which includes at most about three (e.g., one, two, or three) amino acid substitutions, LC-CDR2 comprises the amino acid sequence shown in any of SEQ ID NOs: 65-69 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 65-69, which includes at most about three (e.g., one, two, or three) amino acid substitutions, and LC-CDR3 comprises SEQ ID The amino acid sequence includes one of NOs:70-80 or a variant of the amino acid sequence shown in one of SEQ ID NOs:70-80, which includes at most about three (e.g., one, two, or three) amino acid substitutions. In some examples, the IgG2 is human IgG2. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147.In some embodiments, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0215] In several examples, a full-length anti-CD40 antibody containing an IgG3 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 1-8 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 1-8 comprising at most about three (e.g., one, two, or three) amino acid substitutions, the HC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 9-31 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 9-31 comprising at most about three (e.g., one, two, or three) amino acid substitutions, and the HC-CDR3 comprising SEQ ID The light chain variable domain comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises the amino acid sequence shown in any of SEQ ID NOs: 58-64 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-64, which includes at most about three (e.g., one, two, or three) amino acid substitutions, LC-CDR2 comprises the amino acid sequence shown in any of SEQ ID NOs: 65-69 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 65-69, which includes at most about three (e.g., one, two, or three) amino acid substitutions, and LC-CDR3 comprises SEQ ID The amino acid sequence includes the amino acid sequence shown in any of NOs:70-80 or a variant of the amino acid sequence shown in any of SEQ ID NOs:70-80, which includes at most about three (e.g., one, two, or three) amino acid substitutions. In some examples, the IgG3 is human IgG3. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147.In some embodiments, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0216] In several examples, a full-length anti-CD40 antibody containing an IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 1-8 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 1-8 comprising at most about three (e.g., one, two, or three) amino acid substitutions, the HC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 9-31 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 9-31 comprising at most about three (e.g., one, two, or three) amino acid substitutions, and the HC-CDR3 comprising SEQ ID The light chain variable domain comprises LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises the amino acid sequence shown in any of SEQ ID NOs: 58-64 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 58-64, which includes at most about three (e.g., one, two, or three) amino acid substitutions, LC-CDR2 comprises the amino acid sequence shown in any of SEQ ID NOs: 65-69 or a variant of the amino acid sequence shown in any of SEQ ID NOs: 65-69, which includes at most about three (e.g., one, two, or three) amino acid substitutions, and LC-CDR3 comprises SEQ ID The amino acid sequence includes the amino acid sequence shown in any of NOs:70-80 or a variant of the amino acid sequence shown in any of SEQ ID NOs:70-80, which includes at most about three (e.g., one, two, or three) amino acid substitutions. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146.In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0217] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, or a variant of the heavy chain variable domain, the HC-CDR1 comprising the amino acid sequence shown in any of SEQ ID NOs: 1-8, the HC-CDR2 comprising the amino acid sequence shown in any of SEQ ID NOs: 9-31, and the HC-CDR3 comprising SEQ ID The HC-CDR sequence of the variant of the heavy chain variable domain includes an amino acid sequence shown in any of NOs:32-57, and the HC-CDR sequence of the variant of the heavy chain variable domain includes at most about five (e.g., one, two, three, four, or five) amino acid substitutions, and the light chain variable domain includes LC-CDR1, LC-CDR2, and LC-CDR3, or includes a variant of the light chain variable domain, where LC-CDR1 includes an amino acid sequence shown in any of SEQ ID NOs:58-64, LC-CDR2 includes an amino acid sequence shown in any of SEQ ID NOs:65-69, and LC-CDR3 includes an amino acid sequence shown in any of SEQ ID NOs:70-80, and the LC-CDR sequence of the variant of the light chain variable domain includes at most about five (e.g., one, two, three, four, or five) amino acid substitutions. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147.In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0218] In several examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, or a variant of the heavy chain variable domain, the HC-CDR1 comprising the amino acid sequence shown in any of SEQ ID NOs: 1-8, the HC-CDR2 comprising the amino acid sequence shown in any of SEQ ID NOs: 9-31, and the HC-CDR3 comprising SEQ ID The HC-CDR sequence of the variant of the heavy chain variable domain includes an amino acid sequence shown in any of NOs:32-57, and the HC-CDR sequence of the variant of the heavy chain variable domain includes at most about five (e.g., one, two, three, four, or five) amino acid substitutions, and the light chain variable domain includes LC-CDR1, LC-CDR2, and LC-CDR3, or includes a variant of the light chain variable domain, where LC-CDR1 includes an amino acid sequence shown in any of SEQ ID NOs:58-64, LC-CDR2 includes an amino acid sequence shown in any of SEQ ID NOs:65-69, and LC-CDR3 includes an amino acid sequence shown in any of SEQ ID NOs:70-80, and the LC-CDR sequence of the variant of the light chain variable domain includes at most about five (e.g., one, two, three, four, or five) amino acid substitutions. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147.In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0219] In several examples, a full-length anti-CD40 antibody containing an IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 1-8, the HC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 9-31, the HC-CDR3 comprising an amino acid sequence shown in any of SEQ ID NOs: 32-57, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 58-64, the LC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 65-69, and the LC-CDR3 comprising SEQ ID The amino acid sequence includes one of the NOs:70-80. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0220] In several examples, a full-length anti-CD40 antibody containing an IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 1-8, the HC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 9-31, the HC-CDR3 comprising an amino acid sequence shown in any of SEQ ID NOs: 32-57, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising an amino acid sequence shown in any of SEQ ID NOs: 58-64, the LC-CDR2 comprising an amino acid sequence shown in any of SEQ ID NOs: 65-69, and the LC-CDR3 comprising SEQ ID The amino acid sequence includes one of the NOs:70-80. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0221] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:9, the HC-CDR3 comprising amino acid sequence SEQ ID NO:32, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:58, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0222] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:10, the HC-CDR3 comprising amino acid sequence SEQ ID NO:32, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0223] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:13, the HC-CDR3 comprising amino acid sequence SEQ ID NO:34, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0224] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:15, the HC-CDR3 comprising amino acid sequence SEQ ID NO:37, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:73. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0225] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:20, the HC-CDR3 comprising amino acid sequence SEQ ID NO:42, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0226] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:14, the HC-CDR3 comprising amino acid sequence SEQ ID NO:43, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0227] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:51, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:59, LC-CDR2 comprises amino acid sequence SEQ ID NO:65, and LC-CDR3 comprises amino acid sequence SEQ ID NO:70. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0228] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:11, the HC-CDR3 comprising amino acid sequence SEQ ID NO:48, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:71. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0229] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:18, the HC-CDR3 comprising amino acid sequence SEQ ID NO:47, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:71. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0230] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:4, the HC-CDR2 comprising amino acid sequence SEQ ID NO:27, the HC-CDR3 comprising amino acid sequence SEQ ID NO:53, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:62, the LC-CDR2 comprising amino acid sequence SEQ ID NO:67, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:78. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0231] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO: 5, the HC-CDR2 comprising amino acid sequence SEQ ID NO: 27, the HC-CDR3 comprising amino acid sequence SEQ ID NO: 54, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO: 62, the LC-CDR2 comprising amino acid sequence SEQ ID NO: 67, and the LC-CDR3 comprising amino acid sequence SEQ ID NO: 78. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0232] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO: 6, the HC-CDR2 comprising amino acid sequence SEQ ID NO: 28, the HC-CDR3 comprising amino acid sequence SEQ ID NO: 55, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO: 63, the LC-CDR2 comprising amino acid sequence SEQ ID NO: 68, and the LC-CDR3 comprising amino acid sequence SEQ ID NO: 79. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0233] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:7, the HC-CDR2 comprising amino acid sequence SEQ ID NO:29, the HC-CDR3 comprising amino acid sequence SEQ ID NO:56, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:64, the LC-CDR2 comprising amino acid sequence SEQ ID NO:69, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:80. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0234] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO: 8, HC-CDR2 comprises amino acid sequence SEQ ID NO: 30, and HC-CDR3 comprises amino acid sequence SEQ ID NO: 57, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 62, LC-CDR2 comprises amino acid sequence SEQ ID NO: 67, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 80. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0235] In some examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:4, the HC-CDR2 comprising amino acid sequence SEQ ID NO:31, the HC-CDR3 comprising amino acid sequence SEQ ID NO:53, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:62, the LC-CDR2 comprising amino acid sequence SEQ ID NO:68, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:79. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0236] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:9, the HC-CDR3 comprising amino acid sequence SEQ ID NO:32, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:58, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0237] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:10, the HC-CDR3 comprising amino acid sequence SEQ ID NO:32, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0238] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:13, the HC-CDR3 comprising amino acid sequence SEQ ID NO:34, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:70. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0239] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:37, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:59, LC-CDR2 comprises amino acid sequence SEQ ID NO:65, and LC-CDR3 comprises amino acid sequence SEQ ID NO:73. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0240] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:20, and HC-CDR3 comprises amino acid sequence SEQ ID NO:42, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:59, LC-CDR2 comprises amino acid sequence SEQ ID NO:65, and LC-CDR3 comprises amino acid sequence SEQ ID NO:70. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0241] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:14, and HC-CDR3 comprises amino acid sequence SEQ ID NO:43, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:59, LC-CDR2 comprises amino acid sequence SEQ ID NO:65, and LC-CDR3 comprises amino acid sequence SEQ ID NO:70. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0242] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:1, HC-CDR2 comprises amino acid sequence SEQ ID NO:15, and HC-CDR3 comprises amino acid sequence SEQ ID NO:51, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:59, LC-CDR2 comprises amino acid sequence SEQ ID NO:65, and LC-CDR3 comprises amino acid sequence SEQ ID NO:70. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0243] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:11, the HC-CDR3 comprising amino acid sequence SEQ ID NO:48, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:71. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0244] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:1, the HC-CDR2 comprising amino acid sequence SEQ ID NO:18, the HC-CDR3 comprising amino acid sequence SEQ ID NO:47, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:59, the LC-CDR2 comprising amino acid sequence SEQ ID NO:65, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:71. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0245] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:4, HC-CDR2 comprises amino acid sequence SEQ ID NO:27, and HC-CDR3 comprises amino acid sequence SEQ ID NO:53, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:62, LC-CDR2 comprises amino acid sequence SEQ ID NO:67, and LC-CDR3 comprises amino acid sequence SEQ ID NO:78. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0246] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO: 5, HC-CDR2 comprises amino acid sequence SEQ ID NO: 27, and HC-CDR3 comprises amino acid sequence SEQ ID NO: 54, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 62, LC-CDR2 comprises amino acid sequence SEQ ID NO: 67, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 78. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0247] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO: 6, HC-CDR2 comprises amino acid sequence SEQ ID NO: 28, and HC-CDR3 comprises amino acid sequence SEQ ID NO: 55, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 63, LC-CDR2 comprises amino acid sequence SEQ ID NO: 68, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 79. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0248] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO:7, HC-CDR2 comprises amino acid sequence SEQ ID NO:29, and HC-CDR3 comprises amino acid sequence SEQ ID NO:56, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO:64, LC-CDR2 comprises amino acid sequence SEQ ID NO:69, and LC-CDR3 comprises amino acid sequence SEQ ID NO:80. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0249] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, wherein HC-CDR1 comprises amino acid sequence SEQ ID NO: 8, HC-CDR2 comprises amino acid sequence SEQ ID NO: 30, and HC-CDR3 comprises amino acid sequence SEQ ID NO: 57, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1 comprises amino acid sequence SEQ ID NO: 62, LC-CDR2 comprises amino acid sequence SEQ ID NO: 67, and LC-CDR3 comprises amino acid sequence SEQ ID NO: 80. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0250] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, the anti-CD40 antibody comprising a) a heavy chain variable domain and b) a light chain variable domain, the heavy chain variable domain comprising HC-CDR1, HC-CDR2, and HC-CDR3, the HC-CDR1 comprising amino acid sequence SEQ ID NO:4, the HC-CDR2 comprising amino acid sequence SEQ ID NO:31, the HC-CDR3 comprising amino acid sequence SEQ ID NO:53, the light chain variable domain comprising LC-CDR1, LC-CDR2, and LC-CDR3, the LC-CDR1 comprising amino acid sequence SEQ ID NO:62, the LC-CDR2 comprising amino acid sequence SEQ ID NO:68, and the LC-CDR3 comprising amino acid sequence SEQ ID NO:79. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0251] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (VL ) including the V H This includes an amino acid sequence shown in any of SEQ ID NOs:87-125 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with an amino acid sequence shown in any of SEQ ID NOs:87-125, and the V L This includes the amino acid sequence shown in any of SEQ ID NOs:126-144 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence shown in any of SEQ ID NOs:126-144. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148. In some embodiments, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0252] In several examples, a full-length anti-CD40 antibody containing the IgG2 constant region is provided, and the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the VH This includes an amino acid sequence shown in any of SEQ ID NOs:87-125 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with an amino acid sequence shown in any of SEQ ID NOs:87-125, and the V L This includes the amino acid sequence shown in any of SEQ ID NOs:126-144 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence shown in any of SEQ ID NOs:126-144. In some examples, the IgG2 is human IgG2. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0253] In several examples, a full-length anti-CD40 antibody containing the IgG3 constant region is provided, and the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes an amino acid sequence shown in any of SEQ ID NOs:87-125 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with an amino acid sequence shown in any of SEQ ID NOs:87-125, and the V LThis includes the amino acid sequence shown in any of SEQ ID NOs:126-144 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence shown in any of SEQ ID NOs:126-144. In some examples, the IgG3 is human IgG3. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0254] In several examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes an amino acid sequence shown in any of SEQ ID NOs:87-125 or a mutant having at least about 90% (for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with an amino acid sequence shown in any of SEQ ID NOs:87-125, and the V LThis includes the amino acid sequence shown in any of SEQ ID NOs:126-144 or a variant having at least about 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence shown in any of SEQ ID NOs:126-144. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148. In some embodiments, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148.

[0255] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes the amino acid sequence shown in any of SEQ ID NOs:87-125, and the V LThe amino acid sequence includes one of the SEQ ID NOs: 126-144. In some examples, the IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO: 145. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO: 147. In some examples, the light chain constant region includes or consists of the amino acid sequence SEQ ID NO: 148. In some examples, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO: 145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO: 147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0256] In several examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody has a heavy chain variable domain (V H ) and light chain variable domain (V L ) including the V H This includes the amino acid sequence shown in any of SEQ ID NOs:87-125, and the V LThe amino acid sequence includes one of the SEQ ID NOs: 126-144. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO: 146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO: 147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO: 148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO: 146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO: 147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0257] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:87 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:87, and the V LThis includes amino acid sequence SEQ ID NO:126 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:126. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0258] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:88 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:88, and the V LThis includes amino acid sequence SEQ ID NO:127 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:127. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0259] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:89 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:89, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0260] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:94 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:94, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0261] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:96 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:96, and the V LThis includes amino acid sequence SEQ ID NO:132 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:132. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0262] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:101 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:101, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0263] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:110 or a variant having at least approximately 90% sequence identity with the amino acid sequence SEQ ID NO:110, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0264] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:114 or a mutant having at least approximately 90% sequence identity with the amino acid sequence SEQ ID NO:114, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0265] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:115 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:115, and the V LThis includes amino acid sequence SEQ ID NO:129 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:129. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0266] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:117 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:117, and the V LThis includes amino acid sequence SEQ ID NO:129 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:129. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0267] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:120 or a mutant having at least approximately 90% sequence identity with the amino acid sequence SEQ ID NO:120, and the V LThis includes amino acid sequence SEQ ID NO:139 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:139. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0268] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:121 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:121, and the V LThis includes amino acid sequence SEQ ID NO:140 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:140. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0269] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:122 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:122, and the V LThis includes amino acid sequence SEQ ID NO:141 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:141. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0270] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:123 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:123, and the V LThis includes amino acid sequence SEQ ID NO:142 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:142. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0271] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:124 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:124, and the V LThis includes amino acid sequence SEQ ID NO:143 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:143. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0272] In several examples, a full-length anti-CD40 antibody containing the IgG1 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:125 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:125, and the V LThis includes amino acid sequence SEQ ID NO:144 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:144. In some examples, IgG1 is human IgG1. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:145, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0273] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:87 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:87, and the V LThis includes amino acid sequence SEQ ID NO:126 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:126. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0274] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:88 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:88, and the V LThis includes amino acid sequence SEQ ID NO:127 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:127. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0275] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:89 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:89, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0276] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:94 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:94, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0277] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:96 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:96, and the V LThis includes amino acid sequence SEQ ID NO:132 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:132. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0278] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:101 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:101, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0279] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:110 or a variant having at least approximately 90% sequence identity with the amino acid sequence SEQ ID NO:110, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0280] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:114 or a mutant having at least approximately 90% sequence identity with the amino acid sequence SEQ ID NO:114, and the V LThis includes amino acid sequence SEQ ID NO:128 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:128. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0281] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:115 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:115, and the V LThis includes amino acid sequence SEQ ID NO:129 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:129. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0282] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:117 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:117, and the V LThis includes amino acid sequence SEQ ID NO:129 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:129. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0283] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes the amino acid sequence SEQ ID NO:120 or a mutant having at least approximately 90% sequence identity with the amino acid sequence SEQ ID NO:120, and the V LThis includes amino acid sequence SEQ ID NO:139 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:139. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0284] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:121 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:121, and the V LThis includes amino acid sequence SEQ ID NO:140 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:140. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0285] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:122 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:122, and the V LThis includes amino acid sequence SEQ ID NO:141 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:141. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0286] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:123 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:123, and the V LThis includes amino acid sequence SEQ ID NO:142 or a variant having at least about 90% sequence identity with amino acid sequence SEQ ID NO:142. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0287] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:124 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:124, and the V LThis includes amino acid sequence SEQ ID NO:143 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:143. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0288] In some examples, a full-length anti-CD40 antibody containing the IgG4 constant region is provided, and the anti-CD40 antibody is V H and V L Including the above V H This includes amino acid sequence SEQ ID NO:125 or a mutant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:125, and the V LThis includes amino acid sequence SEQ ID NO:144 or a variant having at least approximately 90% sequence identity with amino acid sequence SEQ ID NO:144. In some examples, the IgG4 is human IgG4. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some examples, the light chain constant region includes or consists of amino acid sequence SEQ ID NO:148. In some examples, the heavy chain constant region includes or consists of amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of amino acid sequence SEQ ID NO:147. In some embodiments, the heavy chain constant region includes or consists of the amino acid sequence SEQ ID NO:146, and the light chain constant region includes or consists of the amino acid sequence SEQ ID NO:148.

[0289] binding affinity Binding affinity is expressed as Kd, Koff, Kon, or Ka. As used herein, the term Koff refers to the rate constant at which an antibody dissociates from an antigen / antibody complex, and is measured by a kinetic sorting device. The term Kon refers to the binding rate constant at which an antibody binds to an antigen to form an antigen / antibody complex. As used herein, the equilibrium dissociation constant Kd refers to the dissociation constant at which a particular antibody interacts with an antigen, and is equal to Koff / Kon, the antigen concentration required to reach equilibrium in an antibody molecule solution where the antigen occupies half of all antibody binding sites. In measuring Kd, it is assumed that all binding molecules are present in solution. When an antibody binds to a cell wall, for example in the yeast phenotype, the corresponding equilibrium dissociation rate constant is a good approximation of Kd, EC 50It is expressed as follows. The affinity coupling constant Ka is the reciprocal of the dissociation constant Kd.

[0290] The dissociation constant (Kd) is used as an indicator of the affinity between the antibody moiety and the antigen. For example, antibodies labeled with various labels using the Scatchard method, and simple analyses can be performed using the Biacore instrument (manufactured by Amershm Biosciences), and the interactions between biomolecules can be analyzed by surface plasmon resonance according to the user manual or the included reagent kit. The Kd values ​​obtained using these methods are expressed in units of M. Antibodies that specifically bind to a target have a Kd value of, for example, ≤10 -7 M, ≤10 -8 M, ≤10 -9 M, ≤10 -10 M, ≤10 -11 M, ≤10 -12 M, or ≤10 -13 It may have a Kd value of M.

[0291] The binding specificity of an antibody can be measured experimentally using methods known in the art. These methods include, but are not limited to, Western blots, ELISA, RIA, ECL, IRMA, EIA, BIAcore tests, and peptide scanning.

[0292] In some examples, the anti-CD40 antibody specifically binds to the CD40 target, and its Kd value is 10 -7 M~10 -13 M (for example, 10 -7 M~10 -13 M, 10 -8 M~10 -13 M, 10 -9 M~10 -13 M, or 10 -10 M~10 -12 M) Therefore, in some examples, the Kd value of binding between the anti-CD40 antibody and CD40 is 10 -7 M~10 -13 M, 1×10 -7 M~5×10 -13 M, 10 -7M~10 -12 M、10 -7 M~10 -11 M、10 -7 M~10 -10 M、10 -7 M~10 -9 M、10 -8 M~10 -13 M、1×10 -8 M~5×10 -13 M、10 -8 M~10 -12 M、10 -8 M~10 -11 M、10 -8 M~10 -10 M、10 -8 M~10 -9 M、5×10 -9 M~1×10 -13 M、5×10 -9 M~1×10 -12 M、5×10 -9 M~1×10 -11 M、5×10 -9 M~1×10 -10 M、10 -9 M~10 -13 M、10 -9 M~10 -12 M、10 -9 M~10 -11 M、10 -9 M~10 -10 M、5×10 -10 M~1×10 -13 M、5×10 -10 M~1×10 -12 M、5×10 -10 M~1×10 -11 M、10 -10 M~10 -13 M、1×10 -10 M~5×10 -13 M、1×10 -10 M~1×10 -12 M、1×10 -10 M~5×10 -12 M、1×10 -10 M~1×10 -11 M、10 -11 M~10 -13 M、1×10 -11 M~5×10 -13 M、10-11 M~10 -12 M, or 10 -12 M~10 -13 M is the value. In some examples, the Kd value of binding between the anti-CD40 antibody and CD40 is 10 -7 M~10 -13 It is M.

[0293] In some examples, the Kd value of binding between the anti-CD40 antibody and the non-target is higher than the Kd value of binding between the anti-CD40 antibody and the target, and in some examples cited herein, the binding affinity between the anti-CD40 antibody and the target (e.g., CD40) is higher than the binding affinity between the anti-CD40 antibody and the non-target. In some examples, the non-target refers to a non-CD40 antigen. In some examples, there is at least a 10-fold difference between the Kd values ​​of binding between the anti-CD40 antibody (against CD40) and the non-CD40 target, for example, 10 to 100 times, 100 to 1000 times, and 10 3 ~10 4 double, 10 4 ~10 5 double, 10 5 ~10 6 double, 10 6 ~10 7 double, 10 7 ~10 8 double, 10 8 ~10 9 double, 10 9 ~10 10 double, 10 10 ~10 11 double, or 10 11 ~10 12 There is a twofold difference.

[0294] In some examples, the Kd value of the binding of the anti-CD40 antibody to the non-target was 10 -1 M~10 -6 M (for example, 10 -1 M~10 -6 M, 10 -1 M~10 -5 M, or 10 -2 M~10 -4M) In some examples, the non-target refers to a non-CD40 antigen. Therefore, in some examples, the Kd value of binding between the anti-CD40 antibody and the non-CD40 target is 10 -1 M~10 -6 M, 1×10 -1 M~5×10 -6 M, 10 -1 M~10 -5 M, 1×10 -1 M~5×10 -5 M, 10 -1 M~10 -4 M, 1×10 -1 M~5×10 -4 M, 10 -1 M~10 -3 M, 1×10 -1 M~5×10 -3 M, 10 -1 M~10 -2 M, 10 -2 M~10 -6 M, 1×10 -2 M~5×10 -6 M, 10 -2 M~10 -5 M, 1×10 -2 M~5×10 -5 M, 10 -2 M~10 -4 M, 1×10 -2 M~5×10 -4 M, 10 -2 M~10 -3 M, 10 -3 M~10 -6 M, 1×10 -3 M~5×10 -6 M, 10 -3 M~10 -5 M, 1×10 -3 M~5×10 -5 M, 10 -3 M~10 -4 M, 10 -4 M~10 -6 M, 1×10 -4 M~5×10 -6 M, 10 -4 M~10 -5 M, or 10 -5 M~10 -6 It is M.

[0295] In some examples, the anti-CD40 antibody specifically recognizes the CD40 target with high binding affinity and binds to non-targets with low binding affinity. In such cases, the Kd value of the binding between the anti-CD40 antibody and the CD40 target is 10. -7 M~10 -13 M (for example, 10 -7 M~10 -13 M, 10 -8 M~10 -13 M, 10 -9 M~10 -13 M, or 10 -10 M~10 -12 M) and the Kd value for binding to the non-target is 10 -1 M~10 -6 M (for example, 10 -1 M~10 -6 M, 10 -1 M~10 -5 M, or 10 -2 M~10 -4 M) is the answer.

[0296] In some examples, when referring to the specific recognition of CD40 by an anti-CD40 antibody, the binding affinity of the anti-CD40 antibody is compared to the binding affinity of a control anti-CD40 antibody (e.g., ASKP1240). In some examples, the Kd value of binding between the control anti-CD40 antibody and CD40 is at least twice the Kd value of binding between the anti-CD40 antibody and CD40 described herein, for example, 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, 10x, 10-100x, 10-100x, 10 3 ~10 4 It could be double.

[0297] nucleic acid Nucleic acid molecules encoding anti-CD40 antibodies are also subject to consideration in this invention. In some examples, a nucleic acid (or set of nucleic acids) encoding a full-length anti-CD40 antibody is provided, comprising one of the full-length anti-CD40 antibodies described herein. In some examples, the nucleic acid (or set of nucleic acids) of the anti-CD40 antibody described herein may also include a nucleic acid sequence encoding a polypeptide tag (e.g., a protein purification tag, a His tag, or an HA tag).

[0298] Simultaneously, this specification also considers isolated host cells containing an anti-CD40 antibody, isolated nucleic acids encoding an anti-CD40 antibody polypeptide component, or vectors containing nucleic acids encoding an anti-CD40 antibody polypeptide component as described herein.

[0299] The present invention also includes variants of these nucleic acid sequences. For example, the variants include nucleotide sequences that hybridize with the nucleic acid sequence encoding the anti-CD40 antibody of the present invention under at least moderate stringent hybridization conditions.

[0300] The present invention also provides a vector into which the nucleic acid sequence described in the present invention can be inserted.

[0301] Simply put, by inserting a native or synthetic nucleic acid encoding an anti-CD40 antibody into a suitable expression vector, the nucleic acid can be operably linked to regulatory elements at the 5' and 3' ends, including, for example, a promoter (e.g., a lymphocyte-specific promoter) and a 3' untranslated region (UTR), thereby expressing an anti-CD40 antibody (e.g., a full-length anti-CD40 antibody). The vector can be applied to replication and integration in eukaryotic host cells. Typical cloning and expression vectors include transcriptional and translational terminators, start sequences, and promoters that regulate the expression of a target nucleic acid sequence.

[0302] The nucleic acids described herein are used in nucleic acid immunotherapy and gene therapy by using standard gene delivery methods. Nucleic acid delivery methods are known in the art. See, for example, USPat. Nos. 5,399,346, 5,580,859, and 5,589,466, which are incorporated herein by reference in their entirety. In some embodiments, the present invention also provides gene therapy vectors.

[0303] Nucleic acids can be cloned into many types of vectors. For example, nucleic acids can be cloned into vectors, which include, but are not limited to, plasmids, phages, phage derivatives, animal viruses, and cosmids. Of particular interest are expression vectors, replication vectors, probe-generating vectors, and sequencing vectors.

[0304] Furthermore, expression vectors can be delivered to cells in the form of viral vectors. Viral vector technology is well known in the art and is described, for example, in Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York) and other virology or molecular biology handbooks. Viruses used as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses. Generally, a suitable vector includes an origin of replication that acts in at least one organism, a promoter sequence, a convenient restriction endonuclease site, and one or more selection markers (see, e.g., WO 01 / 96584; WO 01 / 29058; and USPat. No. 6,326,193).

[0305] Many virus-based systems have been developed to introduce genes into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. By applying techniques known in the art, selected genes can be inserted into vectors and packaged into retroviral particles. The recombinant viruses are then isolated and delivered to the subject's cells in vivo or in vitro. Many retroviral systems are known in the art. Adenovirus vectors are used in some examples. Many adenovirus vectors are known in the art. Lentiviral vectors are used in some examples. For example, vectors derived from retroviruses such as lentiviruses are suitable tools for achieving long-term gene transfer because they allow for the long-term and stable integration of the introduced genes and their proliferation in the next generation of cells. Lentiviral vectors have further advantages over retroviruses derived from tumors, such as mouse leukemia virus, because they can transduce non-dividing cells, such as hepatocytes. At the same time, they also have the further advantage of low immunogenicity.

[0306] For example, other promoter elements, such as enhancers, regulate the transcription initiation frequency. Generally, these are located 30–110 bp upstream of the start site, but recently, many promoters have been found to also include functional elements downstream of the start site. The spacing between promoter elements is usually flexible, so the promoter's function is maintained even if the positions of the elements are swapped or shifted. In the thymidine kinase (TK) promoter, activity begins to decrease only when the spacing between promoter elements increases to 50 bp.

[0307] One example of a suitable promoter is the pre-early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a very strong constitutive promoter sequence and can drive high levels of expression of any operablely linked polynucleotide sequence. Another example of a suitable promoter is the elongation factor 1α (EF-1α) promoter. However, other constitutive promoters can be used, and these other constitutive promoters include, but are not limited to, the Simian virus 40 (SV40) early promoter, mouse mammary cancer virus (MMTV), human immunodeficiency virus long-terminal repeat (HIV-LTR) promoter, MoMLV promoter, avian leukemia virus promoter, Epstein-Barr virus pre-early promoter, Roussarcoma virus promoter, and human gene promoters, and these human gene promoters include, but are not limited to, the actin promoter, myosin promoter, hemoglobin promoter, and creatine kinase promoter. Furthermore, the present invention should not be limited to the use of constitutive promoters only, as inducible promoters are also considered in the present invention. The use of inducible promoters provides a kind of molecular switch that can initiate the expression of a activatably linked polynucleotide sequence when necessary and stop its expression when not needed. Inducible promoters include, but are not limited to, metallothionein promoters, glucocorticoid promoters, progesterone promoters, and tetracycline promoters.

[0308] In some examples, the expression of anti-CD40 antibodies is induceable. In some examples, the nucleic acid sequence encoding the anti-CD40 antibody is operably linked to an inducible promoter, including one of the inducible promoters described herein.

[0309] Induced promoter The use of an inducible promoter provides a molecular switch that can initiate the expression of a activatably linked polynucleotide sequence when expression is needed and stop it when it is not needed. Exemplary inducible promoters suitable for eukaryotic cells include, but are not limited to, hormone-regulated elements (e.g., see Mader, S. and White, JH (1993) Proc. Natl. Acad. Sci. USA 90:5603-5607), synthetic ligand-regulated elements (see Spencer, D. et al. (1993) Science 262:1019-1024), and ionizing radiation-regulated elements (see Manome, Y. et al. (1993) Biochemistry 32:10607-10613; see Datta, R. et al. (1992) Proc. Natl. Acad. Sci. USA 89:1014-10153). For exemplary inducible promoters suitable for other in vivo or in vitro mammalian systems, see Gingrich et al. (1998) Annual Rev. Neurosci 21:377-405. In some examples, the inducible promoter system for expressing anti-CD40 antibody is the Tet system. In some examples, the inducible promoter system for expressing anti-CD40 antibody is the E. coli lac repressor system.

[0310] An exemplary inducible promoter system used in the present invention is the Tet system. This system is based on the Tet system described by Gossen et al. (1993). In exemplary examples, a target polynucleotide is controlled by a promoter containing one or more Tet operator (TetO) sites. In the inactive state, a Tet repressor (TetR) binds to the TetO site and inhibits the transcription of the promoter. In the activated state, in the presence of an inducer such as tetracycline (Tc), anhydrous tetracycline, doxycycline (Dox), or an active analog thereof, the inducer causes the release of TetR from TetO, thereby inducing transcription. Doxycycline is a member of the tetracycline antibiotic family, and its chemical name is 1-dimethylamino-2,4a,5,7-pentahydroxy-11-methyl-4,6-dioxy-1,4a,11,11a,12,12a-hexahydrotetraene-3-formamide.

[0311] In one example, TetR is codon-optimized and suitable for expression in mammalian cells, such as mouse or human cells. Due to the degenerate nature of the genetic code, most amino acids are coded by multiple codons, and while there are many variants of a particular nucleic acid sequence, the coded amino acid sequence remains unchanged. However, many organisms differ in codon usage, which is also called "codon bias" (i.e., a preference for certain amino acids to use certain codons). Codon bias is usually associated with the presence of a dominant tRNA type for a particular codon, which conversely increases the efficiency of mRNA translation. Therefore, coding sequences derived from a specific species (e.g., prokaryotes) can be customized by codon optimization to enhance expression in different species (e.g., eukaryotes).

[0312] Other specific variants of the Tet system include the "Tet-Off" and "Tet-On" systems described below. In the Tet-off system, transcription is inactivated in the presence of Tc or Dox. In this system, a tetracycline-regulated transcription-activating protein (tTA), consisting of a fusion of TetR and the strongly transcription-activating domain of herpes simplex virus VP16, regulates the expression of target nucleic acids under the transcriptional control of a tetracycline-responsive promoter element (TRE). The TRE element consists of a fusion of a TetO sequence series and a promoter (usually a minimal promoter sequence derived from the human cytomegalovirus pre-early promoter). In the absence of Tc or Dox, tTA binds to the TRE and activates the transcription of the target gene. In the presence of Tc or Dox, tTA cannot bind to the TRE, and the target gene cannot be expressed.

[0313] In contrast, in the Tet-On system, transcription is activated in the presence of Tc or Dox. The Tet-On system is based on the reverse tetracycline-regulated transcription activator rtTA. Like tTA, rtTA is a fusion protein consisting of a TetR repressor and a VP16 transcriptional activation domain. However, four amino acid changes in the DNA-binding region of TetR alter the binding properties of rtTA, allowing it to recognize only the tetO sequence of the TRE of the target transgene in the presence of Dox. Therefore, in the Tet-On system, the transcription of target genes regulated by TRE can only be activated in the presence of Dox.

[0314] Another inducible promoter system is the lac repressor system in E. coli (see Brown et al., Cell 49:603-612 (1987)). The lac repressor system functions by regulating the transcription of a target polynucleotide operably linked to a promoter containing a lac operator (lacO). The lac repressor (lacR) inhibits the transcription of the target polynucleotide by binding to lacO. For example, the expression of the target polynucleotide is induced by a suitable inducer such as isopropyl-β-D-thiogalactopyranoside (IPTG).

[0315] To evaluate the expression of a polypeptide or a portion thereof, the expression vector introduced into cells may include a selection marker gene, a reporter gene, or both, for recognizing and selecting cells of interest from a transfected or infected cell population using a viral vector. In other embodiments, the selection marker is supported on a single DNA fragment and used in cotransfection experiments. Both the selection marker gene and the reporter gene can be flanked by appropriate regulatory sequences to enable expression in host cells. Useful selection markers include, for example, antibiotic resistance genes such as neo and similar genes.

[0316] Reporter genes are used to identify potential transfection cells and to evaluate the function of regulatory sequences. Typically, a reporter gene is a gene that is not present in or expressed by the receptor organism or tissue, and encodes a polypeptide to be discovered as a detectable characteristic, such as enzymatic activity. Once the DNA is introduced into the receptor cell, the expression of the reporter gene is detected at the appropriate time. Suitable reporter genes include genes that secrete coded luciferases, β-galactosidases, chloramphenicol acetyltransferases, basic phosphatases, or green fluorescent protein (see Ui-Tel et al., 2000 FEBS Letters 479:79-82). Suitable expression systems are publicly known and can be manufactured by known techniques or commercially available. Generally, a construct of the smallest 5' flanking region capable of exhibiting the highest expression level of the reporter gene is recognized as a promoter. Such promoter regions can bind to the reporter gene and are used to evaluate the ability of a substance to regulate promoter-driven transcription.

[0317] In some examples, nucleic acids encoding any of the full-length anti-CD40 antibodies described herein are provided. In some examples, the nucleic acid comprises one or more nucleic acid sequences encoding the heavy and light chains of the full-length anti-CD40 antibody. In some examples, each of the one or more nucleic acid sequences is contained in a single vector. In some examples, at least several nucleic acid sequences are contained in the same vector. In some examples, all nucleic acid sequences are contained in the same vector. The vector can be selected from, for example, mammalian expression vectors and viral vectors (e.g., vectors derived from retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses).

[0318] Methods for introducing genes into cells and expressing them are known in the art. In the case of expression vectors, the vector can be easily introduced into host cells such as mammalian cells, bacteria, yeast, or insect cells by any method in the art. For example, expression vectors can be introduced into host cells by physical, chemical, or biological methods.

[0319] Physical methods for introducing polynucleotides into host cells include calcium phosphate precipitation, lipofection, particle gun, microinjection, and electroporation. Methods for preparing cells containing vectors and / or exogenous nucleic acids are well known in the art. See, for example, Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). In some examples, polynucleotides are introduced into host cells by calcium phosphate transfection.

[0320] Biological methods for introducing targeted polynucleotides into host cells include the use of DNA and RNA vectors. Viral vectors, particularly retroviral vectors, are the most widely used method for inserting genes into mammalian cells, such as human cells. Other viral vectors can be derived from lentiviruses, poxviruses, herpes simplex virus type 1, adenoviruses, and adeno-associated viruses, among others. See USPat. Nos. 5,350,674 and 5,585,362.

[0321] Chemical methods for introducing polynucleotides into host cells include colloidal dispersion systems such as polymer complexes, nanocapsules, microspheres, and magnetic beads, as well as lipid-based systems including oil-in-water emulsions, colloids, mixed colloids, and liposomes. An exemplary colloidal system used as a delivery vector in vivo and in vitro is liposomes (e.g., artificial membrane sacs).

[0322] When using nonviral delivery systems, an exemplary delivery vector is a liposome. Consider the delivery of nucleic acids to host cells (in vitro, ex vivo, or in vivo) using lipid formulations. In another embodiment, the nucleic acid can be bound to a lipid. The lipid-bound nucleic acid can be encapsulated within the aqueous interior of a liposome, dispersed within the lipid bilayer of a liposome, bound to a liposome via a linking molecule conjugated to the liposome and oligonucleotide, embedded in a liposome, form a complex with a liposome, dispersed in a lipid-containing solution, mixed with a lipid, bound to a lipid, suspended in a lipid, contained in or mixed with a micelle, or otherwise bound to a lipid. Compositions related to lipids, lipid / DNA, or lipid / expression vectors are not limited to a specific structure in solution. For example, they can exist in a bilayer structure, a micelle, or a "depressed" structure. They can also be easily dispersed in solution and may form aggregates of heterogeneous size and shape. Lipids are fatty substances and may be naturally occurring or synthetically produced lipids. For example, lipids include naturally occurring lipid droplets in the cytoplasm, as well as compounds containing long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes.

[0323] Regardless of the method by which exogenous nucleic acids are introduced into host cells, or by other means of exposing cells to the inhibitors of the present invention, various experiments can be performed to confirm the presence of recombinant DNA sequences in the host cells. Such experiments include, for example, “molecular biology” experiments well known to those skilled in the art, such as Southern and Northern blotting, RT-PCR, and PCR; “biochemical” experiments, such as detecting the presence of specific polypeptides, and identification by immunological methods (ELISAs and Western blots) or the experiments described herein, all of which fall within the scope of the present invention.

[0324] Preparation of anti-CD40 antibody In some examples, the anti-CD40 antibody is a monoclonal antibody or derived from a monoclonal antibody. In some examples, the anti-CD40 antibody is derived from a monoclonal antibody. H and V L or variants thereof. In some examples, the anti-CD40 antibody further comprises the CH1 and CL regions derived from a monoclonal antibody, or variants thereof. Monoclonal antibodies can be prepared using methods known in the art, including, for example, the application of hybridoma cell methods, phage display methods, or recombinant DNA methods. Exemplary phage display methods are described herein and in the following examples.

[0325] In the hybridoma cell method, hamsters, mice, or other suitable host animals are generally immunized with an immunotherapy agent to induce lymphocytes that produce or are capable of producing antibodies that specifically bind to the immunotherapy agent. Alternatively, lymphocytes can be immunized in vitro. The immunotherapy agent may include polypeptides of target proteins or fusion proteins. Typically, when human cells are required, peripheral blood lymphocytes (PBLs) are used, while when non-human mammalian cells are required, splenocytes or lymph node cells are used. To form hybridoma cells, lymphocytes are fused with an immortalized cell line using a suitable fusion agent, such as polyethylene glycol. The immortalized cell line is typically transformed mammalian cells, particularly rodent, bovine, and human myeloma cells. Rat or mouse myeloma cell lines are commonly used. Hybridoma cells can be cultured in a suitable medium, which preferably contains one or more substances that inhibit the growth or survival of unfused immortalized cells. For example, if parent cells are deficient in hypoxanthine-guanine phosphoribosyltransferase (HGPRT or HPRT), hybridoma cell culture media typically contain hypoxanthine, aminopterin, and thymidine (HAT medium), which can inhibit the growth of HGPRT-deficient cells.

[0326] In several examples, immortalized cell lines effectively fuse, ensuring stable expression of antibodies at high levels via selected antibody-producing cells, and are sensitive to specific media such as HAT medium. In some examples, the immortalized cell lines are mouse myeloma cell lines, which can be obtained, for example, from the Salk Cell Preservation Center in San Diego, California, and the American Type Culture Collection in Manassas, Virginia. Simultaneously, the production of human-source monoclonal antibodies using human myeloma and mouse-human hybrid myeloma cell lines is also described.

[0327] Next, the presence of monoclonal antibodies against polypeptides in the culture medium for hybridoma cells can be measured. The binding specificity of monoclonal antibodies produced from hybridoma cells can be determined by immunoprecipitation or in vitro binding experiments, such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques or analytical methods are known in the art. The binding affinity of monoclonal antibodies can be determined, for example, by Scatchard analysis described in Munson and Pollard, Anal. Biochem., 107:220 (1980).

[0328] After identifying the required hybridoma cells, subcloning can be performed on target clones using limiting dilution and cultured using standard methods. Suitable media for this purpose include, for example, Dulbecco's Modified Eagle Medium (DMEM) and RPMI-1640 medium. Alternatively, hybridoma cells can be grown in mammalian in vivo in the form of ascites fluid.

[0329] Monoclonal antibodies secreted by subclones can be isolated or purified from culture media or ascites fluid by conventional immunoglobulin purification methods such as protein A-Sepharose chromatography, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0330] In some examples, according to any anti-CD40 antibody described herein, the anti-CD40 antibody comprises a sequence of a clone selected from an antibody library (e.g., scFv, or a phage library displaying Fab fragments). The clone can be identified by a method of screening an antibody fragment combinatorial library having the desired activity. For example, various methods are known in the art for generating phage display libraries and screening these libraries to obtain antibodies with desired binding properties. These methods are summarized, for example, in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001), and also in, for example, McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352:624-628 (1991); Marks et al., J.Mol.Biol.222:581-597 (1992); Marks and Bradbury, Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al., J.Mol.Biol.338(2):299-310 (2004); Lee et al. Further details can be found in al., J.Mol.Biol.340(5):1073-1093(2004); Fellouse, Proc.Natl.Acad.Sci.USA 101(34):12467-12472(2004); and Lee et al., J.Immunol.Methods 284(1-2):119-132(2004).

[0331] In some phage display methods, as described in Winter et al., Ann. Rev. Immunol., 12:433-455 (1994), polymerase chain reaction (PCR) is used to display V H and V LEach component of a gene is cloned and randomly recombined in a phage library, after which phages capable of binding to the antigen are screened. The phages typically display antibody fragments in the form of scFv or Fab fragments. Immunogen library phages provide high-affinity antibodies against immunogens without requiring the construction of hybridoma cells. Alternatively, natural libraries (e.g., human-derived) can be cloned without immunization to provide single antibody sources against multiple non-self and self-antigens, as described in Griffiths et al., EMBO J, 12:725-734 (1993). Finally, natural libraries can also be prepared by cloning stem cell-derived unreconstituted V-gene fragments, encoding the CDR3 hypervariable region using PCR primers containing random sequences, and completing the reconstitution in vitro, as described in Hoogenboom and Winter, J. Mol. Biol., 227:381-388 (1992). Patent documents describing human antibody phage libraries include, for example, USPat. No. 5,750,373, and US Patent Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0332] The anti-CD40 antibody is prepared by a method of screening for anti-CD40 antibody portions in a library that can specifically bind to target CD40 using phage display. This library may be a human scFv phage display library and contains at least 1 × 10⁶ of cells. 9 (For example, at least 1 × 10 9 , 2.5×10 9 , 5×10 9 , 7.5×10 9 , 1 x 10 10 , 2.5×10 10 , 5×10 10, 7.5×10 10 , or 1 × 10 11 The library contains unique human antibody fragments exhibiting a diversity of types. In some examples, the library is a natural human library constructed from DNA extracted from PMBCs and spleens of healthy subjects and includes all human heavy and light chain subfamilies. In some examples, the library is a natural human library constructed from DNA extracted from PMBCs isolated from the bodies of patients with various diseases, such as patients with autoimmune diseases, cancer patients, and patients with infectious diseases. In some examples, the library is a semi-synthetic human library in which the heavy chain CDR3 is completely random and all amino acids (except cysteine) are present at any given position with equal probability. (See, e.g., Hoet, RMet al., Nat. Biotechnol. 23(3):344-348, 2005). In some examples, the heavy chain CDR3 of the semi-synthetic human library is an amino acid with a length of 5 to 24 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24). In some examples, the library is a totally synthetic phage display library. In some examples, the library is a non-human phage display library.

[0333] Phage clones with high affinity for target CD40 can be screened by repeated binding of the phage to the target CD40. The target CD40 is bound to a solid support (e.g., beads for solution panning or mammalian cells for cell panning), after which unbound phages are removed and the specifically binding phages are eluted. The bound phage clones are then eluted and used to infect suitable host cells, such as E. coli XL1-Blue, for expression and purification. Multiple panning cycles (e.g., 2, 3, 4, 5, 6, or more) may be performed to enrich phage clones that specifically bind to CD40, such as solution panning, cell panning, or a combination of both. Specific binding of the enriched phage clones to the target CD40 can be detected by any method known in the art, including, for example, ELISA and FACS.

[0334] Another method for screening antibody libraries is to display proteins on the surface of yeast cells. Wittrup et al. (US Patents 6,699,658 and 6,696,251) developed a method for a yeast cell display library. In this yeast display system, one component contains yeast agglutinin protein (Aga1) immobilized on the yeast cell wall, and the other component contains a second subunit of agglutinin protein Aga2, which can be bound to the Aga1 protein via a disulfide bond and displayed on the surface of the yeast cell. The Aga1 protein is expressed by incorporating the Aga1 gene into the yeast chromosome. After fusing a single-stranded variable fragment (scFv) library with the Aga2 gene in a yeast display plasmid and transforming it, the library can be retained in yeast in the presence of additional nutritional markers. Both Aga1 and Aga2 proteins are expressed under the control of a galactose-inducible promoter.

[0335] Human antibody V gene bank (V H , and V KThe fragments were obtained by PCR using a single set of degenerate primers (Sblattero, D. and Bradbury, A. Immunotechnology 3, 271-278 1998). PCR templates are obtained from commercially available RNA or cDNA, including PBMCs, spleen, lymph nodes, bone marrow, and amygdala. H and V K After integrating the PCR library, the scFv format is assembled by overlap extension PCR (Sheets, MD et al, Proc. Natl. Acad. Sci. USA 95, 6157-6162 1998). To construct a yeast scFv display library, the obtained scFv PCR products are cloned into yeast display plasmids in yeast by homologous recombination (Chao, G, et al, Nat Protoc. 2006; 1(2): 755-68. Miller KD, et al. Current Protocols in Cytometry 4.7.1-4.7.30, 2008).

[0336] Anti-CD40 antibodies can be screened using mammalian cell display systems, where the antibody moiety is displayed on the cell surface and the screening method is antigen-directed, allowing for the isolation of antibodies that specifically target CD40 (as described in USpatent No. 7,732,195B2). A Chinese hamster ovary (CHO) cell library displaying large amounts of human IgG antibody genes has been created and is used to discover clones expressing high-affinity antibody genes. Another display system has been developed that, through alternative splicing, simultaneously displays and secretes the same protein on the cell surface, maintaining a correlation between the displayed protein phenotype and genotype, thereby allowing the secreted soluble antibody to be simultaneously characterized in biophysical and cellular function-based analyses. This method overcomes many of the limitations of conventional mammalian cell displays and allows for the direct screening and maturation of full-length, glycosylated IgG antibodies (Peter M. Bowers, et al, Methods 2014, 65:44-56). Transient expression systems are best suited for single-run antigen selection performed before the antibody gene is restored, making them most useful for selecting antibodies from small libraries. Stable episomal vectors offer an attractive option. Episomal vectors can be efficiently transfected and maintained stably at low copy numbers, enabling multiple panning cycles and analysis of more complex antibody libraries.

[0337] The IgG library is constructed based on the ligation of germline sequence V gene fragments isolated from a group of human donors with the reconstructed (D)J region. RNA taken from 2000 human blood samples is reverse transcribed into cDNA, and V H and V K Using a specific primer, V H and V K The fragments are amplified and purified by gel extraction. H and V KThe fragments are subcloned into display vectors containing either the IgG1 or K constant region, respectively, and then transduced into 293T cells by electroporation or transduction, thereby preparing an IgG library. To prepare the scFv antibody display library, V H and V K The scFv is produced by ligating the fragments, which are then subcloned into a display vector and transduced into 293T cells by electroporation or transduction. As is well known, the IgG library is constructed based on germline sequence V gene fragments isolated from a donor population and a reconstituted (D)J region, the donor may be a mouse, rat, rabbit, or monkey.

[0338] Monoclonal antibodies can also be produced by recombinant DNA methods, such as those described in US Patent No. 4,816,567. The DNA encoding the monoclonal antibodies described in this invention can be readily isolated and sequenced by conventional methods (e.g., by oligonucleotide probes that can specifically bind to the genes encoding the light and heavy chains of mouse-derived antibodies). Hybridoma cells as described above, or the CD40-specific phage clones of this invention, can be used as a source of this DNA. After isolation, the DNA can be placed in an expression vector, and this vector can then be transfected into host cells, such as monkey COS cells, Chinese hamster ovarian cancer (CHO) cells, or myeloma cells that do not produce immunoglobulins, to obtain monoclonal antibodies synthesized in recombinant host cells. The DNA may be modified, for example, by substituting the human heavy and light chain constant regions with coding sequences and / or substituting homologous non-human sequences in the framework region (US Patent No. 4,816,567; Morrison et al., supra), or by covalently linking all or part of the coding sequence of the immunoglobulin with the coding sequence of a non-immunoglobulin polypeptide. Such non-immunoglobulin polypeptides can substitute the constant region of the antibody of the present invention, or substitute one of the antigen-binding sites in the antibody variable domain of the present invention to form a chimeric bivalent antibody.

[0339] The antibody may be a monovalent antibody. Methods for preparing monovalent antibodies are known in the art. For example, recombinant expression methods for immunoglobulin light chains and modified heavy chains. Typically, the heavy chains are cleaved at any position in the Fc region to prevent crosslinking. Alternatively, the relevant cysteine ​​residues are substituted for or deleted with other amino acid residues to prevent crosslinking.

[0340] The in vitro method is also suitable for the preparation of monovalent antibodies. The antibody can be digested to produce antibody fragments, particularly Fab fragments, using any method known in the art.

[0341] An antibody variable domain having the desired binding specificity (antibody-antigen binding site) can be fused to an immunoglobulin constant region. It is preferable to fuse to an immunoglobulin heavy chain constant region containing at least some hinge, CH2, and CH3 regions. In some examples, the first heavy chain constant region (CH1), which contains the sites necessary for light chain binding, is present in at least one fusion. The DNA encoding the immunoglobulin heavy chain fusion may, if necessary, include DNA encoding the immunoglobulin light chain, be inserted into an independent expression vector, and cotransfected into a suitable host organism.

[0342] Fully human antibodies and humanized antibodies The anti-CD40 antibody (e.g., full-length anti-CD40 antibody) may be a fully human antibody or a humanized antibody. The humanized form of the non-human (e.g., mouse) antibody portion is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (e.g., Fv, Fab, Fab', F(ab')2, scFv, or other antigen-binding subsequence of the antibody), and typically includes the smallest sequence derived from the non-human immunoglobulin. The humanized antibody includes a human immunoglobulin, immunoglobulin chain, or fragment thereof (receptor antibody), and the residues of the receptor CDR are replaced with non-human (donor antibody) CDR residues, such as mouse, rat, or rabbit CDRs, which have the desired specificity, affinity, and properties. In some examples, human immunoglobulin Fv framework region residues are replaced with corresponding non-human residues. The humanized antibody may also include amino acid residues that are neither receptor antibodies nor present in the introduced CDR or framework region sequence. Generally, humanized antibodies contain at least one, usually two, variable domains, with all or nearly all CDR regions corresponding to the CDR regions of non-human immunoglobulins, and all or nearly all framework regions being the consensus sequence of human immunoglobulins.

[0343] Generally, humanized antibodies contain one or more amino acid residues introduced from a non-human source. These non-human amino acid residues are generally called “implant” residues and usually originate from an “implant” variable domain. According to several examples, humanization can usually be performed by the following methods of Winter and colleagues (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by substituting the corresponding sequence of the human antibody with rodent CDRs or CDR sequences. Thus, this “humanized” antibody moiety (USPatent No. 4, 816, 567) is usually less than that of a complete human antibody, with its variable domain being substituted by a corresponding sequence of non-human origin. In reality, the humanized antibody portion is a typical human antibody portion, with some CDR residues and several possible framework region residues being substituted with residues from similar sites in rodent antibodies.

[0344] Fully human antibodies are an alternative method of humanization. For example, it is now possible to prepare transgenic animals (e.g., mice) that can produce a complete fully human antibody library without producing endogenous immunoglobulins after immunization. For instance, homozygous deletion of the antibody heavy chain binding region (JH) gene in chimeric and germline mutant mice has been reported to completely inhibit the production of endogenous antibodies. Human antibodies can be produced under antigen stimulation by transferring human germline immunoglobulin gene sequences into the bodies of this type of germline mutant mice, see, for example, Jakobovits et al., PNAS USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immunol., 7:33 (1993); US Patent Nos. 5,545,806,5,569,825,5,591,669,5,545,807; and WO 97 / 17852. Alternatively, fully human antibodies can be prepared by introducing human immunoglobulin loci into transgenic animals (e.g., mice in which the endogenous immunoglobulin gene is partially or completely inactivated). Following antigen stimulation, the production of fully human antibodies was found to be remarkably similar to that in humans in various aspects, including gene rearrangement, assembly, and antibody library management.This method is described, for example, in USPatent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016, and Marks et al., Bio / Technology, 10:779-783 (1992); Lonberg et al., Nature, 368:856-859 (1994); Morrison, Nature, 368:812-813 (1994); Fishwild et al., Nature Biotechnology, 14:845-851 (1996); Neuberger, Nature Biotechnology, 14:826 (1996); Lonberg and Huszar, Intern. Rev. Immunol., 13:65-93 (1995).

[0345] Fully human antibodies can also be produced by activating B cells in vitro (see USPatents 5,567,610 and 5,229,275) or by various techniques known in the art, including phage display libraries. The techniques of Hoogenboom and Winter, J.Mol.Biol., 227:381 (1991); Marks et al., J.Mol.Biol., 222:581 (1991); Cole et al., and Boerner et al. are also used for the preparation of fully human monoclonal antibodies. See Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p.77 (1985) and Boerner et al., J.Immunol., 147(1):86-95 (1991).

[0346] Mutants of anti-CD40 antibodies In some examples, the amino acid sequences of anti-CD40 antibody variants (e.g., full-length anti-CD40 antibodies) provided herein are also considered. For example, it may be necessary to improve the antibody's binding affinity and / or other biological activity. The amino acid sequences of antibody variants can be produced by introducing appropriate modifications to the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, the deletion and / or insertion and / or substitution of residues in the antibody amino acid sequence. The final construction can be achieved by any combination of deletion, insertion, and substitution of amino acid residues to have the desired characteristics, for example, antigen-binding properties.

[0347] In several examples, anti-CD40 antibody variants having one or more amino acid substitutions are provided. Target sites for substitution mutations include hypervariable regions (HVRs) and framework regions (FRs). By introducing amino acid substitutions into the target antibody, it is possible to screen for desired active products having, for example, improved biological activity, maintained / improved antigen-binding ability, reduced immunogenicity, or improved ADCC or CDC.

[0348] Conservative substitutions are shown in Table 4.

[0349] [Table 4]

[0350] Amino acids are classified into different types based on the properties of their side chains: a. Hydrophobic amino acids: norleucine, methionine, alanine, valine, leucine, isoleucine; b. Neutral hydrophilic amino acids: cysteine ​​(Cys), serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln); c. Acidic amino acids: Aspartic acid (Asp), Glutamic acid (Glu); d. Basic amino acids: histidine (His), lysine (Lys), arginine (Arg); e. Amino ...

Claims

1. An isolated anti-CD40 antibody or antigen-binding fragment comprising any of the following (i) to (ix): (i) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 9, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 32, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 58, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 70, (ii) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 10, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 32, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 70, (iii) V Ｈ and V Ｌ comprising The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 13, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 34, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 70, (iv) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 15, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 37, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 73, (v)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 includes the amino acid sequence SEQ ID NO: 20, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 42, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 70, (vi)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 14, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 43, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 70, (vii) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 15, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 51, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 70, (viiii)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 11, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 48, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 71, (ix)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ This includes HC-CDR1, HC-CDR2, and HC-CDR3. The aforementioned HC-CDR1 contains the amino acid sequence SEQ ID NO: 1, The aforementioned HC-CDR2 contains the amino acid sequence SEQ ID NO: 18, The aforementioned HC-CDR3 contains the amino acid sequence SEQ ID NO: 47, The aforementioned V Ｌ This includes LC-CDR1, LC-CDR2, and LC-CDR3. The aforementioned LC-CDR1 contains the amino acid sequence SEQ ID NO: 59, The aforementioned LC-CDR2 contains the amino acid sequence SEQ ID NO: 65, The aforementioned LC-CDR3 contains the amino acid sequence SEQ ID NO: 71, Isolated anti-CD40 antibody or antigen-binding fragment.

2. An isolated anti-CD40 antibody or antigen-binding fragment comprising any of the following (i) to (x): (i) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 87, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 126, (ii) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 88, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 127, (iii) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 89, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 128, (iv) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 94, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 128, (v)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 96, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 132, (vi)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 101, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 128, (vii) V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 110, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 128, (viiii)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 114, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 128, (ix)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 115, The aforementioned V Ｌ It contains amino acid sequence SEQ ID NO: 129, (x)V Ｈ and V Ｌ Includes, The aforementioned V Ｈ It contains amino acid sequence SEQ ID NO: 117, The aforementioned V Ｌ This includes amino acid sequence SEQ ID NO: 129, The isolated anti-CD40 antibody or antigen-binding fragment according to claim 1.

3. The isolated anti-CD40 antibody or antigen-binding fragment according to claim 1, wherein the anti-CD40 antibody or antigen-binding fragment comprises an Fc fragment.

4. The isolated anti-CD40 antibody or antigen-binding fragment according to claim 3, wherein the anti-CD40 antibody or antigen-binding fragment is a full-length IgG antibody.

5. The isolated anti-CD40 antibody or antigen-binding fragment according to claim 4, wherein the anti-CD40 antibody or antigen-binding fragment is a full-length IgG1 antibody, IgG2 antibody, IgG3 antibody, or IgG4 antibody.

6. The isolated anti-CD40 antibody or antigen-binding fragment according to any one of claims 1 to 5, wherein the anti-CD40 antibody or antigen-binding fragment is a chimeric antibody or a humanized antibody.

7. The antigen-binding fragments are Fab, Fab', and F(ab)'. ２ An isolated anti-CD40 antibody or antigen-binding fragment according to claim 1, selected from Fab'-SH, a single-chain antibody (scFv), an Fv fragment, a diabody, and a linear antibody.

8. A nucleic acid molecule encoding an isolated anti-CD40 antibody or antigen-binding fragment as described in claim 1.

9. A vector comprising the nucleic acid molecule described in claim 8.

10. Isolated host cells comprising an isolated anti-CD40 antibody or antigen-binding fragment according to claim 1, a nucleic acid molecule according to claim 8, or a vector according to claim 9.

11. a) Culturing the host cells described in claim 10 under conditions in which an anti-CD40 antibody or antigen-binding fragment can be effectively expressed, and b) Obtaining expressed anti-CD40 antibodies or antigen-binding fragments from host cells. A method for preparing an anti-CD40 antibody or antigen-binding fragment, including the above.

12. A pharmaceutical composition comprising an isolated anti-CD40 antibody or antigen-binding fragment according to claim 1, a nucleic acid molecule according to claim 8, or a vector according to claim 9, and a pharmaceutically acceptable carrier.

13. Use of the isolated anti-CD40 antibody or antigen-binding fragment according to claim 1, the nucleic acid molecule according to claim 8, or the vector according to claim 9 in the preparation of a drug for treating a target disease or condition.

14. The use according to claim 13, wherein the disease or condition is an autoimmune disease, an inflammatory disease, cancer, and / or a transplant-related disease.

15. The aforementioned diseases or conditions include systemic lupus erythematosus (SLE), discoid lupus erythematosus, lupus nephritis, inflammatory arthritis, organ or tissue transplant rejection, multiple sclerosis, hyper-IgE syndrome, polyarteritis nodosa, primary biliary cirrhosis, inflammatory bowel disease, Crohn's disease, celiac disease (gluten-sensitive bowel syndrome), autoimmune hepatitis, pernicious anemia, autoimmune hemolytic anemia, scleroderma, myasthenia gravis, idiopathic thrombocytopenic purpura, autoimmune thyroiditis, Graves' disease, and Hashimoto's thyroiditis. Phyroiditis, immune complex disease, chronic fatigue immune dysfunction syndrome (CFIDS), polymyositis, dermatomyositis, cryoglobulinemia, thrombolysis, cardiomyopathy, pemphigus vulgaris, interstitial pulmonary fibrosis, type 1 and type 2 diabetes, type 1, type 2, type 3 and type 4 delayed-type hypersensitivity, allergies or allergic diseases, unnecessary / unintentional immune responses to therapeutic proteins, asthma, Rich-Strauss syndrome (allergic granulomatosis), atopic dermatitis, allergic contact dermatitis, irritant contact dermatitis, urticaria, IgE-mediated allergy, atherosclerosis, vasculitis, idiopathic inflammatory muscle disease, hemolytic disease, Alzheimer's disease, chronic inflammatory demyelinating polyneuropathy, Sjögren's syndrome The use according to claim 13, selected from disease, psoriasis, hidradenitis suppurativa, urethritis suppurativa, sarcoidosis, ulcerative colitis, tumor, nephritis, respiratory distress syndrome, epididymitis, glomerulonephritis, and hyperthyroidism.

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