WO2024148208A1

WO2024148208A1 - Antibodies specifically recognizing tnfr2 and compositions and uses thereof

Info

Publication number: WO2024148208A1
Application number: PCT/US2024/010380
Authority: WO
Inventors: Zuoan YI; Wenwu Zhai; Chong HE
Original assignee: Staidson Biopharma Inc.
Priority date: 2023-01-06
Filing date: 2024-01-05
Publication date: 2024-07-11

Abstract

The present application relates to a composition of anti-TNFR2 antibody, wherein the composition comprises an isolated anti-TNFR2 antibody in an amount of from about Img/ml to about 300mg/ml, a stabilizer, a surfactant and a buffering agent. The compositions of the application have good stability under various conditions, e.g., freeze-thaw, high temperature and long-term conditions (2- 8 °C and 25 °C), which can ensure the compositions maintain good stability during preparation, transportation and storage process, and ensure the safety and quality control of clinical drugs.

Description

ANTIBODIES SPECIFICALLY RECOGNIZING TNFR2 AND COMPOSITIONS AND

USES THEREOF

REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0001] This application claims the benefit of U.S. Provisional Application No. 63/478,907, filed on January 6, 2023, the content of which is incorporated herein by reference in its entirety. The contents of the electronic sequence listing (ANTI-TNFR2 ANTIBODY AND COMPOSITIONS SEQLIST.xml; Size: 11 bytes; and Date of Creation: Dec 13, 2023) is herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0002] This application pertains to antibodies that specifically recognize tumor necrosis factor receptor 2 (TNFR2), and compositions, and methods of treating diseases or conditions mediated through TNFR2, such as cancer or infectious diseases.

BACKGROUND OF THE INVENTION

[0003] Tumor necrosis factor receptor 2 (TNFR2), also known as tumor necrosis factor receptor superfamily member IB (TNFRSF1B) and CD 120b, is a membrane receptor that binds with cognate ligand TNFa and also with lymphotoxin-a (LTα). In contrast to TNFR1 which has a death domain (DD) in its cytoplasmic part and activates caspase-dependent pathway and NFKB pathway, TNFR2 lacks DD but can recruit the adapter protein TNF receptor associated factor 2 (TRAF2) and TRAF3 and activates the nonconical NFKB pathway and MAP kinase pathway (Brenner et al., 2015). TNFR2 is expressed on the immune cells and some non-immune cells including endothelial cells, cardiomyocytes, astrocytes, etc. (Ward-Kavanagh et al., 2016). Although early studies showed that TNFR2 co-stimulates naive T cell function, it was later demonstrated that TNFR2 also limits CD8+ T-cell-mediated viral clearance and anti-tumor immunity by inducing rapid contraction of CD8+ T cells (Bertrand et al., 2015; DeBerge et al., 2015; Kim et al., 2009; Wortzman et al., 2013b). Several studies showed that TNFR2 expression is higher in regulatory T cells (Treg cells) than in naive T cells and TNFR2 signaling is important for the development, proliferation, and survival of Treg cells (Chen et al., 2013; Horwitz et al., 2013; Mahmud et al., 2014). Therefore, TNFR2 signaling plays critical roles in regulating immune response. In addition, TNFR2 is highly expressed in Treg cells and myeloid-derived suppressive cells (MDSC) in tumor microenvironment, indicating the potential function of TNFR2 in tumor immunity (Chen et al., 2013; Hu et al., 2014). In fact, several publications reported the antitumor efficacy of anti-mouse TNFR2 antibody, although the mechanisms remain ill-defined (Case et al., 2020; Nie et al., 2016; Tam et al., 2019; Williams et al., 2018). Furthermore, an identified mutation in TNFR2 was linked to T cell lymphoma, including mycosis fungoides and Sezary syndrome, suggesting it may serve as an oncogene (Ungewickell et al., 2015). There are also reports showing that TNFR2 is upregulated in certain types of cancer and related with poor prognosis (Yang et al., 2017; Zhang et al., 2018). [0004] Thus, there remains a need in the art for therapeutic antibodies that effectively inhibit or otherwise antagonize TNFR2, and related composition thereof, and methods of treating diseases or conditions mediated through TNFR2, such as cancer or infectious diseases.

[0005] The disclosures of all publications, patents, patent applications and published patent applications referred to herein are hereby incorporated herein by reference in their entirety.

DESCRIPTION OF THE INVENTION

[0006] One aspect of the present application provides a composition, The composition comprises an isolated anti-TNFR2 antibody in an amount of from about Img/ml to about 300mg/ml, a stabilizer, a surfactant and a buffering agent.

[0007] In some embodiments according to any one of the compositions described above, the antibody is in an amount of from about 15 mg/ml to about 180 mg/ml, preferably, from about 40 mg/ml to about 60 mg/ml. hi some embodiments, the antibody is in an amount of lmg/ml, lOmg/ml, 15 mg/ml ,20 mg/ml, 40 mg/ml, 45mg/ml, 50mg/ml, 55mg/ml, 60mg/ml, 80mg/ml, 100mg/ml, 120 mg/ml, 140 mg/ml, 160 mg/ml, 180 mg/ml, 200 mg/ml, 220 mg/ml, 250 mg/ml or 300 mg/ml.

[0008] In some embodiments according to any one of the compositions described above, the stabilizer is any one of sucrose, trehalose, maltose, sorbitol, mannitol, sodium chloride, arginine, glycine, proline or lysine, or a combination thereof.

[0009] In some embodiments according to any one of the compositions described above, the amount of sodium chloride, arginine, glycine, proline and/or lysine is from about 50 mM to about 300mM, preferably from about 90 mM to about 150mM, more preferably from about 90 mM to about 130mM; and/or the amount of sucrose, trehalose, maltose, sorbitol, and/or mannitol is from about 5 mg/ml to about 80 mg/ml, preferably from about 10 mg/ml to about 30 mg/ml. In some embodiments, the amount of sodium chloride, arginine, glycine, proline or lysine is 50mM, 60mM,70mM, 80mM, 90mM, 95mM, 100mM, 105mM, 110 mM, 115mM, 120mM, 125mM, 130mM, 140mM, 150 mM, 160 mM , 170 mM,180 mM , 200 mM, 220 mM , 250 mM or 300mM; the amount of sucrose, trehalose, maltose, sorbitol, mannitol is 5mg/ml, 8 mg/ml, lOmg/ml, 15 mg/ml, 20 mg/ml, 25mg/ml, 30mg/ml, 40 mg/ml, 50 mg/ml, 60 mg/ml, 70 mg/ml or 80 mg/ml. [0010] In some embodiments according to any one of the compositions described above, (1) the stabilizer are sodium chloride and sucrose, trehalose, maltose, sorbitol and/or mannitol; or (2) the stabilizer is arginine, glycine, proline and/or lysine.

[0011] In some embodiments according to any one of the compositions described above, (1) the stabilizer are sodium chloride and trehalose; or (2) the stabilizer is arginine.

[0012] In some embodiments according to any one of the compositions described above, (1) the stabilizer are sodium chloride in an amount of from about 90 mM to about 130mM, and sucrose, trehalose, maltose, sorbitol and/or mannitol in an amount of from about lOmg/ml to about 30mg/ml; or (2) the stabilizer is arginine in an amount of from about 110 mM to about 150mM.

[0013] In some embodiments according to any one of the compositions described above, the buffering agent is any one of histidine-histidine hydrochloride buffer, phosphate buffer and aceto- sodium acetate buffer, or a combination thereof; preferably histidine-histidine hydrochloride buffer. [0014] In some embodiments according to any one of the compositions described above, the buffering agent is in an amount of from about 3 mM to about 100 mM; preferably from about 5 mM to about 50 mM; more preferably from about 10 mM to about 30 mM. hi some embodiments, the buffering agent is in an amount of 3mM, 8mM, lOmM, 15mM, 20mM, 25mM, 30mM, 40mM, 50mM, 60mM, 70mM, 80mM, 90mM, 100mM.

[0015] In some embodiments according to any one of the compositions described above, the surfactant is in an amount of from about 0.01mg/ml to about 1.0mg/ml; preferably, from about 0.05mg/ml to about 0.2mg/ml. In some embodiments, the surfactant is in an amount of 0.01mg/ml, 0.05mg/ml, 0.08, mg/ml, 0.1mg/ml, 0.15mg/ml, 0.2mg/ml, 0.5 mg/ml, 0.8 mg/ml or 1.0 mg/ml.

[0016] In some embodiments according to any one of the compositions described above, the surfactant is polysorbitol and/or poloxam; preferably, the polysorbate is Tween-20 or Tween-80. [0017] In some embodiments according to any one of the compositions described above, the composition has a pH of 4.5-6.5, preferably 4.8-6.2. hi some embodiments, the composition has a pH of 4.5, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2 or 6.5.

[0018] In some embodiments according to any one of the compositions described above, the antibody is in an amount of from about Img/ml to about 300mg/ml, preferably from about 15 mg/ml to about 180 mg/ml, more preferably, from about 40 mg/ml to about 60 mg/ml; the stabilizer (1) are sodium chloride in an amount of from about 50 mM to about 300mM, preferably from about 90 mM to about 150mM , more preferably from about 90 mM to about 130mM, and trehalose in an amount of from about 5 mg/ml to about 80 mg/ml, preferably from about lOmg/ml to about 30mg/ml, or (2) the stabilizer is arginine in an amount of from about 50 mM to about 300mM, preferably from about 110 mM to about 150mM; the buffering agent is histidine-histidine hydrochloride buffer in an amount of from about 3 mM to about 100 mM; preferably from about 5 mM to about 50 mM; more preferably from about 10 mM to about 30 mM; the surfactant is Tween-20 or Tween-80 in an amount of from about O.Olmg/ml to about LOmg/ml; preferably, from about 0.05mg/ml to about 0.2mg/ml; the composition has a pH of from 4.5-6.5, preferably 4.8-6.2.

[0019] In some embodiments according to any one of the compositions described above, wherein:

(1) the antibody is in amount of about 40mg/ml, 45mg/ml, 50mg/ml, 55mg/ml or 60 mg/ml; the stabilizer is sodium chloride in an amount of from about 90 mM to about 130mM, and sucrose, trehalose and/or mannitol in an amount of from about lOmg/ml to about 30mg/ml, or the stabilizer is arginine in an amount of from about 110 mM to about 150mM; the buffering agent is histidine- histidine hydrochloride buffer, phosphate buffer and/or aceto-sodium acetate buffer in an amount of from about 10 mM to about 30 mM; the composition has a pH of from about 4.8 to about 6.2; the surfactant is Tween-20 and/or Tween-80 in an amount of from about 0.05mg/ml to about 0.2mg/ml;

(2) the antibody is in amount of from about 40 mg/ml to about 60 mg/ml; the stabilizer is sodium chloride in an amount of about 90mM, 95mM, 100mM, 105mM, 110 mM, 115mM, 120mM, 125mM or 130mM, and sucrose, trehalose and/or mannitol in an amount of from about lOmg/ml, 15 mg/ml, 20 mg/ml, 25mg/ml or 30 mg/ml, or the stabilizer is arginine in an amount of about

1 lOmM, 120mM, 125mM, 130mM, 135 mM, 140mM, 145mM or 150mM; the buffering agent is histidine-histidine hydrochloride buffer, phosphate buffer and/or aceto-sodium acetate buffer in an amount of from about 10 mM to about 30 mM; the composition has a pH of from about 4.8 to about 6.2; the surfactant is Tween-20 and/or Tween-80 in an amount of from about 0.05mg/ml to about 0.2mg/ml;

(3) the antibody is in amount of from about 40 mg/ml to about 60 mg/ml; the stabilizer is sodium chloride in an amount of from about 90 mM to about 130mM, and sucrose, trehalose and/or mannitol in an amount of from about lOmg/ml to about 30mg/ml, or the stabilizer is arginine in an amount of from about 110 mM to about 150mM; the buffering agent is histidine-histidine hydrochloride buffer, phosphate buffer and/or aceto-sodium acetate buffer in an amount of about 10 mM, 20mM or 30 mM; the composition has a pH of from about 4.8 to about 6.2; the surfactant is Tween-20 and/or Tween-80 in an amount of from about 0.05mg/ml to about 0.2mg/ml;

(4) the antibody is in amount of from about 40 mg/ml to about 60 mg/ml; the stabilizer is sodium chloride in an amount of from about 90 mM to about 130mM, and sucrose, trehalose and/or mannitol in an amount of from about lOmg/ml to about 30mg/ml, or the stabilizer is arginine in an amount of from about 110 mM to about 150mM; the buffering agent is histidine-histidine hydrochloride buffer, phosphate buffer and/or aceto-sodium acetate buffer in an amount of from about 10 mM to about 30 mM; the composition has a pH of about 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.06.1, or 6.2; the surfactant is Tween-20 and/or Tween-80 in an amount of from about 0.05mg/ml to about 0.2mg/ml; or,

(5) the antibody is in amount of from about 40 mg/ml to about 60 mg/ml; the stabilizer is sodium chloride in an amount of from about 90 mM to about 130mM, and sucrose, trehalose and/or mannitol in an amount of from about lOmg/ml to about 30mg/ml, or the stabilizer is arginine in an amount of from about 110 mM to about 150mM; the buffering agent is histidine-histidine hydrochloride buffer, phosphate buffer and/or aceto-sodium acetate buffer in an amount of from about 10 mM to about 30 mM; the composition has a pH of from about 4.8 to about 6.2; the surfactant is Tween-20 and/or Tween-80 in an amount of about 0.05mg/ml, 0.08 mg/ml, 0. Img/ml, 0.15mg/ml or 0.2mg/ml.

[0020] In some embodiments according to any one of the compositions described above, wherein:

(1) the antibody is in amount of about 40 mg/ml, the stabilizer is sodium chloride in an amount of about 130mM and trehalose in an amount of about lOmg/ml, the buffering agent is histidine- histidine hydrochloride buffer in an amount of about 10 mM, the composition has a pH of about 6.2; the surfactant is Tween-80 in an amount of about 0.2mg/ml;

(2) the antibody is in amount of about 50 mg/ml, the stabilizer is sodium chloride in an amount of about 1 lOmM and trehalose in an amount of about 20mg/ml, the buffering agent is histidine- histidine hydrochloride buffer in an amount of about 20 mM, the composition has a pH of about 5.7; the surfactant is Tween-20 in an amount of about 0. Img/ml;

(3) the antibody is in amount of about 60 mg/ml, the stabilizer is sodium chloride in an amount of about 90mM and trehalose in an amount of about 30mg/ml, the buffering agent is histidine-histidine hydrochloride buffer in an amount of about 30 mM, the composition has a pH of about 4.8; the surfactant is Tween-20 in an amount of about 0.05mg/ml;

(4) the antibody is in amount of about 50 mg/ml, the stabilizer is sodium chloride in an amount of about 100mM and trehalose in an amount of about 20mg/ml, the buffering agent is aceto-sodium acetate buffer in an amount of about 20 mM, the composition has a pH of about 4.8; the surfactant is Tween-20 in an amount of about 0. Img/ml;

(5) the antibody is in amount of about 55 mg/ml, the stabilizer is sodium chloride in an amount of about 115mM and sucrose in an amount of about 20mg/ml, the buffering agent is phosphate buffer in an amount of about 15 mM, the composition has a pH of about 6.2; the surfactant is Tween-80 in an amount of about 0.2mg/ml; (6) the antibody is in amount of about 45 mg/ml, the stabilizer is sodium chloride in an amount of about 120mM and mannitol in an amount of about 20mg/ml, the buffering agent is phosphate buffer in an amount of about 20 mM, the composition has a pH of about 6.0; the surfactant is Tween-20 in an amount of about 0. Img/ml;

(7) the antibody is in amount of about 40 mg/ml, the stabilizer is arginine in an amount of about 150mM, the buffering agent is histidine-histidine hydrochloride buffer in an amount of about 10 mM, the composition has a pH of about 4.8; the surfactant is Tween-20 in an amount of about 0.2mg/ml;

(8) the antibody is in amount of about 50 mg/ml, the stabilizer is arginine in an amount of about 135mM, the buffering agent is histidine-histidine hydrochloride buffer in an amount of about 20 mM, the composition has a pH of about 5.7; the surfactant is Tween-80 in an amount of about 0. Img/ml;

(9) the antibody is in amount of about 60 mg/ml, the stabilizer is arginine in an amount of about 1 lOmM, the buffering agent is histidine-histidine hydrochloride buffer in an amount of about 30 mM, the composition has a pH of about 6.2; the surfactant is Tween-80 in an amount of about 0.05mg/ml;

(10) the antibody is in amount of about 50 mg/ml, the stabilizer is arginine in an amount of about 140mM, the buffering agent is aceto-sodium acetate buffer in an amount of about 20 mM, the composition has a pH of about 5.0; the surfactant is Tween-80 in an amount of about 0. Img/ml; or

(11) the antibody is in amount of about 45 mg/ml, the stabilizer is arginine in an amount of about 145mM, the buffering agent is phosphate buffer in an amount of about 20 mM, the composition has a pH of about 6.0; the surfactant is Tween-20 in an amount of about 0. Img/ml.

[0021] In some embodiments according to any one of the compositions described above, the composition further comprises a preservative and/or an antioxidant, preferably, the preservative is ethylenediamine tetraacetic acid and/or diethylenetriamine pentaacetic acid; the antioxidant is methionine.

[0022] In some embodiments according to any one of the compositions described above, the preservative is in an amount of from about 0 mg/ml to about 0.5 mg/ml; the antioxidant is in an amount of from about 0 mg/ml to about 2.5 mg/ml.

[0023] In some embodiments according to any one of the compositions described above, the anti- TNFR2 antibody comprises: a heavy chain variable domain (VH) comprising: a heavy chain complementarity determining region (HC-CDR)! comprising the amino acid of SEQ ID NO: 1, and an HC-CDR2 comprising the amino acid of SEQ ID NO: 2; and an HC-CDR3 comprising the amino acid of SEQ ID NO: 3; and a ligjit chain variable domain (VL) comprising: a light chain complementarity determining region (LC-CDR)! comprising the amino acid of SEQ ID NO: 4; a LC-CDR2 comprising the amino acid of SEQ ID NO: 5; and a LC-CDR3 comprising the amino acid of SEQ ID NO: 6.

[0024] In some embodiments according to any one of the compositions described above, the anti- TNFR2 antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 7, 8, 9, 10, 11 or 12; and a VL comprising the amino acid sequence of SEQ ID NO: 13.

[0025] In some embodiments according to any one of the compositions described above, the anti- TNFR2 antibody further comprises: a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 14 or 15 and a ligfit chain constant region comprising the amino acid sequence of SEQ ID NO: 16.

[0026] In some embodiments according to any one of the compositions described above, the anti- TNFR2 antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 7, 9 or 11; and a VL comprising the amino acid sequence of SEQ ID NO: 13, the antibody further comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 14 and a light chain constant region comprising the amino acid sequence of SEQ ID NO: 16; or the anti-TNFR2 antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 8, 10 or 12; and a VL comprising the amino acid sequence of SEQ ID NO: 13, the antibody further comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 15 and a light chain constant region comprising the amino acid sequence of SEQ ID NO: 16. [0027] In some embodiments according to any one of the compositions described above, the composition is liquid or lyophilized, preferably, the composition is sterile.

[0028] Another aspect of the present application provides a method of treating a disease or condition in an individual in need thereof, comprising administering to the individual an effective amount of any one of the compositions described above; preferably, the disease or condition is a cancer or infectious disease, more preferably the disease or condition is associated with TNFR2 signaling or aberrant TNFR2 expression.

[0029] In some embodiments according to the method described above, the disease or condition is selected from non-small cell lung cancer, adrenal gland cancer, bladder cancer, brain cancer, pancreatic adenocarcinoma, breast cancer, colorectal cancer, melanoma, gastroesophageal junction adenocarcinoma, esophageal cancer, esophageal adenocarcinoma, gall bladder cancer, gastric cancer, cervical cancer, gastric adenocarcinoma, head and neck cancer, heart cancer, hepatocellular carcinoma, kidney cancer, liver cancer, mesothelioma, ovarian cancer, pancreatic cancer, prostate cancer, prostate adenocarcinoma, spleen cancer, small cell lung cancer, testicular cancer, thyroid cancer, uterine cancer, and infectious diseases, including, but not limited to Human Papilloma Virus (HPV), Human Immunodeficiency Virus (HIV), Herpes Simplex Virus (HS V), Varicella Zoster Virus (VS V), Cytomegalovirus (CMV), Epstein Barr Virus (EBV), E. coli, Salmonella, Shigella, Staphylococcus aureus, Coliform Bacteria, Chlamydia, Mycobacterium Tuberculosis, Streptococcus, Pneumococcus, Pseudomonas, Campylobacter, Salmonella, Aspergillus Fumigatus, Aspergillus flavus, Cryptococcus Neoformans, and Histoplasma Capsulatum.

[0030] The anti-TNFR2 antibody compositions can be administered to an individual (such as human) via various routes, including, for example, intravenous, intra-arterial, intraperitoneal, intrapulmonary, oral, inhalation, intravesicular, intramuscular, intra-tracheal, subcutaneous, intraocular, intrathecal, transmucosal or transdermal. hi some embodiments, sustained continuous release formulation of the composition may be used. In some embodiments, the composition is administered inhaled. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered intraportally. In some embodiments, the composition is administered intraarterially. In some embodiments, the composition is administered intraperitoneally. In some embodiments, the composition is administered intrahepatically. hi some embodiments, the composition is administered by hepatic arterial infusion, hi some embodiments, the administration is to an injection site distal to a first disease site.

[0031] The term “antibody” herein is used in the broadest sense and encompasses a variety of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, monospecific, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies and antigen-binding fragments thereof, so long as they exhibit the desired antigen binding activity. 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 both chains generally contain three highly variable loops called the complementarity determining regions (CDRs) (light chain (LC) CDRs including LC-CDR1, LC-CDR2, and LC-CDR3, heavy chain (HC) CDRs including HC-CDR1, HC-CDR2, and HC-CDR3). CDR boundaries for the antibodies and antigen-binding fragments disclosed herein may be defined or identified by the conventions of Rabat, Chothia, or Al-Lazikani (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Rabat 1987; Rabat 1991). The three CDRs of the heavy or ligjit chains are interposed between flanking stretches known as framework regions (FRs), which are more highly conserved than the CDRs and form a scaffold to support the hypervariable loops. The constant regions of the heavy and light chains are not involved in antigen binding, but exhibit various effector functions. Antibodies are assigned to classes based on the amino acid sequence of the constant region of their heavy chain. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, which are characterized by the presence of α, δ, ε, γ, and p heavy chains, respectively. Several of the major antibody classes are divided into subclasses such as IgGl (yl heavy chain), IgG2 (y2 heavy chain), IgG3 (y3 heavy chain), IgG4 (y4 heavy chain), IgAl (al heavy chain), or IgA2 (a2 heavy chain). [0032] The term “antigen-binding fragment” as used herein includes an antibody fragment including, for example, a diabody, a Fab, a Fab’, a F(ab’)2, an Fv fragment, a disulfide stabilized Fv fragment (dsFv), a (dsFv)2, a bispecific dsFv (dsFv-dsFv’), a disulfide stabilized diabody (ds diabody), a single-chain Fv (scFv), an scFv dimer (bivalent diabody), a multispecific antibody formed from a portion of an antibody comprising one or more CDRs, a camelized single domain antibody, a nanobody, a domain antibody, a bivalent domain antibody, or any other antibody fragment that binds to an antigen but does not comprise a complete antibody structure. An antigen- binding fragment also includes a fusion protein comprising the antibody fragment described above. An antigen-binding fragment is capable of binding to the same antigen to which the parent antibody or a parent antibody fragment (e.g., a parent scFv) binds. In some embodiments, an antigen-binding fragment may comprise one of more CDRs from a particular human antibody grafted to a framework region from one or more different human antibodies.

[0033] As used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

BRIEF DESCRIPTION OF THE DRAWINGS

[0034] FIG. 1 A shows the binding assay results of the exemplary humanized anti-TNFR2 antibodies as analyzed by FACS. FIG. IB shows the ligand blocking assay results of the exemplary humanized anti-TNFR2 antibodies as analyzed by FACS.

[0035] FIG. 2A shows the tumor volume of individual mouse in different treatment groups. FIG. 2B shows the Inhibition Rate (%) of the average tumor volume in different treatment groups.

EXAMPLES

[0036] The examples below are intended to be purely exemplary of the invention and should therefore not be considered to limit the invention in any way. The following examples and detailed description are offered by way of illustration and not by way of limitation. Example 1: Generation and Characterization of Anti-TNFR2 Antibodies

[0037] This example illustrates the methods of generating anti-TNFR2 antibodies, and methods to screen and select antibodies for further characterization.

[0038] Immunizations and fusions: Balb/c and NZB mice were immunized with recombinant

ECD of human TNFR2 fused with His- or mouse IgG2a Fc produced in Expi293 or CHO cells adjuvanted with RIBI (Sigma Aldrich, cat# S6322-1VL), Titermax (Sigma Aldrich, cat# T2684- 1ML), or/and Freund’s (Freund’s adjuvant, incomplete) (Sigma Aldrich, cat# F5506-10x-10mL). Three days after the last immunization, spleens and lymph nodes were harvested and processed according to standard protocols. Mouse B cells were isolated by EasySep Mouse B cell isolation Kit (StemCell, cat# 19854A) and fused with myeloma cells SP2/0-Agl4 cells (ATCC, CRL 1581) using PEG. Following standard protocols, the fused cells were plated into six-well plates in semi- solid ClonalCell-HY Cloning-Medium D (StemCell, cat# 03804). Monoclonal hybridoma clones were picked into 96 well/plate using Clone Pix 2 Machine (Molecular Devices) and cultured in HT medium.

[0039] Hybridoma Screening: After 10-14 days of culture, supernatants were collected and subjected to primary screening by ELISA with 96 well ELISA plates coated with human or cynomolgus monkey TNFR2 extracellular domain proteins with His or human Fc tag. The parental hybridoma hits identified from the primary screen were expanded. The supernatants of the hybridoma hits identified in the primary screen were further tested for their ability to block the biochemical binding between human TNFR2 and human TNFa. The hybridomas screened were prioritized for subcloning and further characterization.

[0040] Purification of hybridoma antibodies: Positive clone 51B5 was scaled up and the antibody was purified using protein A resin. Then the positive clone 51B5 was sequenced and amplified.

[0041] Generation of recombinant IgGl version of 51B5 chimeric antibody: the recombinant 51B5 chimeric antibody construct with heavy and ligjit chain variable domain of mouse antibody and human constant regions were made by using methods well known in the field. The exemplary human heavy chain constant region and light chain constant region were shown in Table x. The binding affinity and the activity of TNFa binding blocking of 51B5 chimeric antibody was determined by the experiments described below.

Preparation of humanized versions of the hybridoma clone 51B5

[0042] The light chain variable region (VL) and heavy chain variable region (VH) sequences of murine antibody from hybridoma 51B5 were aligned to human germline antibody sequences respectively. The human germline kappa light chain and heavy chain were used as the human frameworks. [0043] The complementarity-determining regions (CDRs) of murine TNFR2 antibody 51B5 light chain and heavy chain were grafted into the identified closest human frameworks respectively to generate humanized antibody clone. In this process, antibody 51B5 was humanized by grafting the CDRs from the murine antibody V-regions onto human germline antibody V-region frameworks, the CDRs grafted from the donor to the acceptor sequence are as defined by Kabat (Kabat et al., 1987). In order to recover the activity of the antibody, a number of framework residues from the murine V-regions that were found to be parts of VH-VL interacting interface or the framework residues acting as “Vernier” zone, which may adjust CDR structure and fine-tune to fit to antigen (Foote et al., 1992) were also retained in the humanized sequence.

[0044] The sequences of the humanized antibodies (e.g., SB1901-72, SB 1901-74, SB1901-76,

SB1901-78, SB 1901-80, or SB 1901-82) were summarized in Table 1-2.

Table 1.

Table 2. Exemplary sequences.

Example 2: In vitro Assays of Humanized Anti-TNFR2 Antibodies

[0045] The full-length IgG versions of humanized TNFR2 antibodies were generated.

Binding affinities

[0046] Binding affinities (monovalent Kd) of anti-TNFR2 antibodies were determined using Biolayer interferometry on the Octet RED96 instrument (ForteBio) at 30 °C and with 1200 rpm agitation. The following kinetic assay was performed using anti-human IgG Fc capture (AHC) biosensors (ForteBio) in kinetics buffer (PBS, 0.1% Tween-20 and 1% bovine serum albumin): (a) antibody (2 μg/mL) for 300 sec, (b) baseline for 120 sec, (c) association with His-tag-huTNFR2 (2.5, 0.5 and 0 μg/mL) for 420 sec and (d) dissociation for 1200 sec. Data fitting and analysis was performed with Octet data analysis software 8.0 (ForteBio) using a 1:1 binding model after Savitzky-Golay filtering. The dissociation constant (Kd) was calculated as the ratio of Koff/Kon. Examples of binding affinity of humanized antibodies (eg. SB1901-72 and SB1901-80) were shown in Table 3.

TABLE 3: Binding affinity of humanized antibodies to TNFR2 antigens

In vitro binding and blocking assays of humanized antibodies

[0047] Binding to huTNFR2 expressed cells

[0048] To examine the binding of anti-TNFR2 antibodies to huTNFR2 expressed cells, Expi293 cells stably expressing huTNFR2 were used to perform FACS analysis.

[0049] The coding sequence of huTNFR2 (Uniprot, P20333) was cloned into a lentiviral vector and the virus was packaged according to the instruction of the virus packaging kit (Lenti-X™ Packaging Single Shots, Cat# 631275, Takada). The Expi293 cells were transduced with the recombinant virus and selected by puromycin. The cell line stably expressing huTNFR2 was incubated with anti-TNFR2 antibodies in PBS with 0.5% BSA, 1 mM EDTA, and 0.1% sodium azide (FACS buffer) for 30 minutes at 4oC. The cells were washed, and then incubated with lOnM phycoerythrin (PE) conjugated anti-Human Fc Ab (Biolegend, cat# 409304) for 20 minutes at 4oC. Cells were washed and then isolated by flow cytometry with Attune (ThermoFisher Scientific).

Data were analyzed with FlowJo software. Antibody binding is represented as median fluorescence intensity (MFI).

[0050] Binding to huTNFR2-Expi293 cells and blocking of TNFa binding

[0051] Expi293 cells stably expressing huTNFR2 were incubated with anti-TNFR2 antibodies for 30 minutes at 4 °C. The cells were washed, and then incubated with 10 nM Alexa Fluor 647 conjugated (ThermoFisher Scientific, cat# A20186) human TNFa (SinoBiological, cat# 10602- HNAE) for 20 minutes at 4 °C . Cells were washed and acquired by flow cytometry with Attune. Data were analyzed with FlowJo software. TNFa binding is represented as MFI.

[0052] . As shown in Figure 1 A and IB, the TNFR2 antibody bound to Expi293-TNFR2 cells and inhibited soluble TNFa binding to TNFR2-expressing Expi293 cells dose-dependently and potently. Example 3: In vivo Anti-tumor Efficacy Study to Evaluate Activity of Humanized Antl- TNFR2 Antibody

[0053] This example illustrated in vivo tumor model study of the functional activity of the humanized anti-TNFR2 antibody.

[0054] A subcutaneous tumor model of MC38 was used here, and the study summary was shown in Table 4 (below).

TABLE 4: Summary of in vivo subcutaneous tumor study protocol

[0055] Animals and husbandry: Forty female C57BL/6-Tnfrsflbtml (TNF-RSF1B)/Bcgen mice (6-9 weeks of age) were used in the studies. The animals were fed breeding diet for “SPF rat and mouse growth” and water ad libitum. Animals were ear tagged for identification purposes and shaved on the left dorsal flank area in preparation of cell implantation. Animals were housed in polycarbonate cages (cage size of 320 x 200 x 135 mm). The environment was controlled to a temperature range of 20°C~26°C and a humidity range of 40%-70%. Animal care and use were compliant with the SOPs of JOINN LABORATORIES (Suzhou) Inc., the Guide for the Care and Use of Laboratory Animals (Sth Edition, Institute of Laboratory Animal Resources, Commission on Life Sciences, National Research Council; National Academy Press; Washington, D.C., 2010), and the U.S. Department of Agriculture through the Animal Welfare Act (Public Law 99-198).

[0056] Cell preparation and implantation: Mouse colon cancer cell line MC38, purchased from Institute of Basic Medical Sciences were cultured and expanded in RPMI medium with 2mM L- glutamine, 10% fetal bovine serum (FBS), and 1% 100x Penicillin/Streptomycin (PS). The growth environment was maintained in an incubator with a 5% CO2 atmosphere at 37°C. When expansion was complete, the cells (passage 3) were trypsinized using a 0.25% trypsin-EDTA solution. The cells were then washed and counted. Pre-implantation cell viability was 92%-94%. The cells were suspended in Dulbecco’s Phosphate Buffered Saline (DPBS) at a concentration of 1x10⁷/ml. Test animals were sterilized at the implantation site with an alcohol prep pad and were implanted subcutaneously in 0.2 mL using a 25-gauge needle and 1 mL syringe.

[0057] Measurements and antibody treatment: Tumors were allowed to grow and mice were then randomized into study groups. Mice were distributed to ensure that the mean body weights for all groups were within 10% of the overall mean tumor burden for the study population. Human MOPC21 IgGl isotype antibody (see Hamlyn PH, Gait MJ, Milstein C. (1981) Complete sequence of an immunoglobulin mRNA using specific priming and the dideoxynucleotide method of RNA sequencing. Nucleic Acids Res. 9(18):4485-4494) and SB1901-72 were made in-house. Mice received twice weekly i.p. injections of each antibody treatment for 3 weeks and tumor volumes were monitored (n=10 mice/ group). The major axis and minor axis of tumors were measured with a vernier caliper and recorded to calculate the tumor volume, and the tumor growth curve was drawn according to the tumor volume to compare the differences between the groups. Tumor volume was calculated according to the following formula: V=1/2xmajor axisxminor axis2. The tumor volume inhibition rate was calculated according to relative tumor volume (RTV) and relative tumor proliferation rate T/C (%).

RTV = Vt/VO

Vt: Tumor volume obtained from each tumor measurement.

V0: Initial tumor volume (before the first dose).

T/C (%) = Mean RTV of Test article group/Mean RTV of control group x 100%.

Tumor volume inhibition rate IRTV (%) = 100% -T/C (%).

[0058] Assessment of side effects: All animals were observed for clinical signs of distress or toxicity at least once daily. Animals were weighed once per week. Animals were euthanized if body weight loss was in excess of 20% or other clinical signs that warranted euthanasia. Individual animals were euthanized when their tumor volume reached or exceeded 2500 mm .

[0059] Results: The tumor volume changes and the mean tumor volume inhibition rate (IRTV, %) in each group were shown in figures 2A and 2B. It is showed that the tumor volume in group 1 was significantly lower than that in isotype antibody treated group (P < 0.001) on Day 22, indicating that TNFR2 blockade by SB1901-72 efficiently suppressed tumor growth. There were no significant differences in body weights between the groups at the same time point. The animals did not show abnormality in general clinical observation.

Example 4: Preparation of anti-TNFR2 antibody formulations

[0060] The formulations of anti-TNFR2 antibody are shown in Table 5 (below). TABLE 5: Formulations of anti-TNFR2 antibody

[0061] Formulations of SB1901-72, SB1901-74(IgGl Fc mutant), SB190 -76, SB1901-78(IgGl Fc mutant), SB1901-80, SB1901-82(IgGl Fc mutant) 1-11 were prepared accordingto Table 5, wherein, SB1901-72, SB 1901-76, SB1901-80 comprise a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 14 and a lighit chain constant region comprising the amino acid sequence of SEQ ID NO: 16; SB1901-74(IgGl Fc mutant), SB1901-78(IgGl Fc mutant), SB1901-82(IgGl Fc mutant) comprise a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 15 and a light chain constant region comprising the amino acid sequence ofSEQ ID NO: 16. Example 5: Stability study of anti-TNFR2 antibody formulations under various conditions

[0062] This study was conducted to test the formulations prepared in Example 4 under a period of exposure to freeze-thaw, high temperature, and long-term conditions. Testing panels included appearance, protein concentration, pH, main peak% (by SEC, Size Exclusion Chromatography), main peak%, acidic peak% and basic peak%cIEF(by Capillary Isoelectric Focusing), intact%(by NR-CE-SDS).

1. Stability study under the condition of Freeze-thaw cycle

[0063] The formulations were shaken at 300 RPM in ambient conditions for up to 36 hours and subjected to freeze-thaw cycling for up to 5 cycles and the stability of the formulation were tested. [0064] (1) Results of the formulations of SB1901-72 are shown in Table 6. Based on the results, all formulations were clear and free of visible particles after 5 freeze-thaw cycles (5FT). There were no significant differences among these formulations after 5FT compared with OFT as analyzed by turbidity, protein concentration, SEC and non-reduced cGE. All of the formulations showed a minor decrease (<0.3 %) in % main peak by SEC and a minor decrease (about 0.1-0.5%) in Intact% by NR-cGE.

[0065] (2) Results of the formulations of SB1901-74(IgGl Fc mutant), SB1901-76, SB1901- 78(IgGl Fc mutant), SB1901-80, SB1901-82(IgGl Fc mutant) also exhibited good stability: all formulations were clear and free of visible particles after 5 freeze-thaw cycles (5FT). There were no significant differences among these formulations after 5FT compared with OFT as analyzed by turbidity, protein concentration, SEC and non-reduced cGE. All of the formulations showed a minor decrease (<0.4 %) in % main peak by SEC and a minor decrease (about 0.1-0.6%) in Intact% by NR-cGE (results are not shown).

[0066] The results indicated that the anti-TNFR2 antibody formulations exhibited good stability under freeze-thaw conditions, for at least 5 freeze-thaw cycles.

Table 6. Results of formulations under freeze-thaw conditions

2. Stability study under the condition of 40°C stressed

[0067] Formulations were filtered using a 0.22 μm membrane, filled in microtubes, sealed with parafilm and incubated at 40°C for up to 4 weeks and the stability of the formulation were tested. [0068] (1) Results of the formulations of SB1901-72 are shown in Table 7. Based on the results, all formulations were clear and free of visible particles over the period of 4-weeks at 40°C. There were also no significant differences among the formulations after 4-weeks at 40°C compared with 0 week. All of the formulations showed a minor decrease (about 3.0-5.8%) in % main peak by SEC and a minor decrease (about 1.5-2.4%) in Intact% by NR-cGE.

[0069] (2) Results of the formulations of SB1901-74(IgGl Fc mutant), SB1901-76, SB1901- 78(IgGl Fc mutant), SB1901-80, SB1901-82(IgGl Fc mutant) also exhibited good stability: all formulations were clear and free of visible particles over the period of 4-weeks at 40°C. There were also no significant differences among the formulations after 4-weeks at 40°C compared with 0 week. All of the formulations showed a minor decrease (about 2.9-6.0%) in % main peak by SEC and a minor decrease (about 1.6-2.8%) in Intact% by NR-cGE (results are not shown).

[0070] The results indicated that the anti-TNFR2 antibody formulations also exhibited good stability under 40°C in at least 4 weeks. Moreover, comparatively speaking, formulation 1-3 and 7- 9 exhibited better stability than any other formulations did.

Table 7. Results of formulations under 40°C stressed

3. Long-term stability test

[0071] 3ml of the formulation 1 -3 or 7-9 prepared in example 5 was filled using aseptic techniques in a 5 mL PETG bottles respectively and stored below 25 "C or at 2-8 "C for 6 months. Then, the stability of the formulations was tested.

Long-term stability test at 2-8 °C

[0072] (1) Results of the formulations of SB1901-72 are shown in Table 8. For that being stored at 2-8°C for 6 months, all formulations were clear and free of visible particles. There were also no significant differences among the formulations after 6 months at 2-8°C compared with 0 month. All of the formulations showed a minor decrease (<0.1%) in % main peak by SEC, a minor decrease (about 1.0- 1.6%) in % main peak, an increase (about 0.6-1.2%) in % acidic peak and an increase (about 0.3-0.5%) in % basic peak by Cief, and a minor decrease (about 0.5-0.9%) in Intact% by NR-cGE.

[0073] (2) Results of the formulations of SB1901-74(igGl Fc mutant), SB1901-76, SB1901- 78(IgGl Fc mutant), SB1901-80, SB1901-82(IgGl Fc mutant) also exhibited good stability: forthat being stored at 2-8°C for 6 months, all formulations were clear and free of visible particles. There were also no significant differences among the formulations after 6 months at 2-8°C compared with 0 month. All of the formulations showed a minor decrease (<0.2%) in % main peak by SEC, a minor decrease (about -0.9- 1.5%) in % main peak, an increase (about 0.6-1.4%) in % acidic peak and an increase (about 0.2-0.7%) in % basic peak by Cief, and a minor decrease (about 0.5-1.1%) in Intact% by NR-cGE (results are not shown).

[0074] The results indicated that the anti-TNFR2 antibody formulations also exhibited good stability after 6 months at 2-8°C. Moreover, comparatively speaking, formulation 1-3 and 7-9 exhibited better stability than any other formulations did.

[0075] Lone-term stability test at 25 °C

[0076] (1) Results of the formulations of SB 1901-72 are shown in Table 9. For that being stored at 25°C for 6 months, all formulations were clear and free of visible particles. There were also no significant differences among the formulations after 6 months at 25°C compared with 0 month. All of the formulations showed a minor decrease (about 1.8-2.1%) in % main peak by SEC, a minor decrease (about 3.7-4.5%) in % main peak , an increase (about 4.8-5.5%) in % acidic peak and a decrease (about 1.0- 1.2% ) in % basic peak by cIEF, and a minor decrease (about 0.2-0.5%) in Intact% by NR-cGE.

[0077] (2) Results of the formulations of SB1901-74(IgGl Fc mutant), SB1901-76, SB1901- 78(IgGl Fc mutant), SB1901-80, SB1901-82(IgGl Fc mutant) also exhibited good stability: forthat being stored at 25°C for 6 months, all formulations were clear and free of visible particles. There were also no significant differences among the formulations after 6 months at 25°C compared with 0 month. All of the formulations showed a minor decrease (about 1.7-2.0%) in % main peak by SEC, a minor decrease (about 3.8-4.8%) in % main peak , an increase (about 4.5-5.7%) in % acidic peak and a decrease (about 1.1-1.4% ) in % basic peak by cIEF, and a minor decrease (about 0.2- 0.8%) in Intact% by NR-cGE (results are not shown).

[0078] The results indicated that the anti-TNFR2 antibody formulations also exhibited good stability after 6 months at 25°C. Moreover, comparatively speaking, formulation 1-3 and 7-9 exhibited better stability than any other formulations did.

Table 8 Results of formulations at 2-8 °C in 6 months

Table 9 Results of formulations at 25*C in 6 months

[0079] The above studies show that the anti-TNFR2 antibody compositions of the application have good stability under various conditions, e.g., freeze-thaw, high temperature and long-term conditions (2-8 "C and 25 *C), which can ensure the compositions maintain good stability during preparation, transportation and storage process, and ensure the safety and quality control of clinical drugs.

[0080] Although the invention has been described in detail with general description, specific implementation method, and examples above, based on the present invention, those skilled in the art can make modifications and improvements within the scope and not deviating from the main purpose of the invention, and these modifications or improvements also belong to the protection scope of the invention.

Claims

1. A composition of anti-TNFR2 antibody, wherein the composition comprises an isolated anti- TNFR2 antibody in an amount of from about Img/ml to about 300mg/ml, a stabilizer, a surfactant and a buffering agent

2. The composition of claim 1, wherein the antibody is in an amount of from about 15 mg/ml to about 180 mg/ml, preferably, from about 40 mg/ml to about 60 mg/ml.

3. The composition of claim 1 or 2, wherein the stabilizer is any one of sucrose, trehalose, maltose, sorbitol, mannitol, sodium chloride, arginine, glycine, proline or lysine, or a combination thereof.

4. The composition of claim 3, wherein the amount of sodium chloride, arginine, glycine, proline and/or lysine is from about 50 mM to about 300mM, preferably from about 90 mM to about 150mM, more preferably from about 90 mM to about 130mM; and/or the amount of sucrose, trehalose, maltose, sorbitol and/or mannitol is from about 5 mg/ml to about 80 mg/ml, preferably from about 10 mg/ml to about 30mg/ml.

5. The composition of any one of claims 1-4, wherein

(1) the stabilizers are sodium chloride and sucrose, trehalose, maltose, sorbitol and/or mannitol; or

(2) the stabilizer is/are arginine, glycine, proline and/or lysine.

6. The composition of claim 5, wherein

(1) the stabilizers are sodium chloride and trehalose; or

(2) the stabilizer is arginine.

7. The composition of claim 5 or 6, wherein

(1) the stabilizers are sodium chloride in an amount of from about 90 mM to about 130mM, and sucrose, trehalose, maltose, sorbitol and/or mannitol in an amount of from about lOmg/ml to about 30mg/ml; or

(2) the stabilizer is arginine in an amount of from about 110 mM to about 150mM.

8. The composition of any one of claims 1-7, wherein the buffering agent is any one of histidine- histidine hydrochloride buffer, phosphate buffer and aceto-sodium acetate buffer, or a combination thereof; preferably histidine-histidine hydrochloride buffer.

9. The composition of any one of claims 1-8, wherein the buffering agent is in an amount of from about 3 mM to about 100 mM; preferably from about 5 mM to about 50 mM; more preferably from about 10 mM to about 30 mM.

10. The composition of any one of claims 1-9, wherein the surfactant is in an amount of from about O.Olmg/ml to about LOmg/ml; preferably, from about 0.05mg/ml to about 0.2mg/ml.

11. The composition of any one of claims 1-10, wherein the surfactant is polysorbitol and/or poloxam; preferably, the polysorbate is Tween-20 or Tween-80.

12. The composition of any one of claims 1-11, wherein the composition has a pH of 4.5-6.S, preferably 4.8-6.2.

13. The composition of claim 1, wherein the antibody is in an amount of from about Img/ml to about 300mg/ml, preferably from about 15 mg/ml to about 180 mg/ml, more preferably, from about 40 mg/ml to about 60 mg/ml; the stabilizer (1) are sodium chloride in an amount of from about 50 mM to about 300mM, preferably from about 90 mM to about 150mM, more preferably, from about 90 mM to about 130mM, and trehalose in an amount of from about 5 mg/ml to about 80 mg/ml, preferably from about lOmg/ml to about 30mg/ml, or (2) is arginine in an amount of from about 50 mM to about 300mM, preferably from about 110 mM to about 150mM; the buffering agent is histidine-histidine hydrochloride buffer in an amount of from about 3 mM to about 100 mM; preferably from about 5 mM to about 50 mM; more preferably from about 10 mM to about 30 mM; the surfactant is Tween-20 or Tween-80 in an amount of from about O.Olmg/ml to about LOmg/ml; preferably, from about 0.05mg/ml to about 0.2mg/ml; the composition has a pH of from 4.5-6.S, preferably 4.8-6.2.

14. The composition of any one of claims 1-13, wherein the composition further comprises a preservative and/or an antioxidant, preferably, the preservative is ethylenediamine tetraacetic acid and/or diethylenetriamine pentaacetic acid; the antioxidant is methionine.

15. The composition of claim 14, wherein the preservative is in an amount of from about 0 mg/ml to about 0.5 mg/ml; the antioxidant is in an amount of from about 0 mg/ml to about 2.5 mg/ml.

16. The composition of any one of claims 1-15, wherein the anti-TNFR2 antibody comprises: a heavy chain variable domain (VH) comprising: a heavy chain complementarity determining region (HC-CDR) 1 comprising the amino acid of SEQ ID NO: 1, and an HC-CDR2 comprising the amino acid of SEQ ID NO: 2; and an HC-CDR3 comprising the amino acid of SEQ ID NO: 3; and a light chain variable domain (VL) comprising: a light chain complementarity determining region (LC-CDR) 1 comprising the amino acid of SEQ ID NO: 4; a LC-CDR2 comprising the amino acid of SEQ ID NO: 5; and a LC-CDR3 comprising the amino acid of SEQ ID NO: 6.

17. The composition of claiml6, wherein the anti-TNFR2 antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 7, 8, 9, 10, 11 or 12; and a VL comprising the amino acid sequence of SEQ ID NO: 13, preferably, the anti-TNFR2 antibody further comprises: a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 14 or 15 and a light chain constant region comprising the amino acid sequence of SEQ ID NO: 16.

18. The composition of claim 16 or 17, wherein, the anti-TNFR2 antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 7, 9 or 11; and a VL comprising the amino acid sequence of SEQ ID NO: 13, the antibody further comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 14 and a ligjit chain constant region comprising the amino acid sequence of SEQ ID NO: 16; or the anti-TNFR2 antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 8, 10 or 12; and a VL comprising the amino acid sequence of SEQ ID NO: 13, the antibody further comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 15 and a ligjit chain constant region comprising the amino acid sequence of SEQ ID NO: 16.

19. The composition of any one of claims 1-18, wherein the composition is liquid or lyophilized, preferably, the composition is sterile.

20. A method of treating a disease or condition in an individual in need thereof, comprising administering to the individual an effective amount of the composition of any one of claims 1-19; preferably, the disease or condition is a cancer or infectious disease, more preferably the disease or condition is associated with TNFR2 signaling or aberrant TNFR2 expression.

21. The method of claim 20, wherein the disease or condition is selected from non-small cell lung cancer, adrenal gland cancer, bladder cancer, brain cancer, pancreatic adenocarcinoma, breast cancer, colorectal cancer, melanoma, gastroesophageal junction adenocarcinoma, esophageal cancer, esophageal adenocarcinoma, gall bladder cancer, gastric cancer, cervical cancer, gastric adenocarcinoma, head and neck cancer, heart cancer, hepatocellular carcinoma, kidney cancer, liver cancer, mesothelioma, ovarian cancer, pancreatic cancer, prostate cancer, prostate adenocarcinoma, spleen cancer, small cell lung cancer, testicular cancer, thyroid cancer, uterine cancer, and infectious diseases, including, but not limited to Human Papilloma Virus (HPV), Human Immunodeficiency Virus (HIV), Herpes Simplex Virus (HS V), Varicella Zoster Virus (VS V), Cytomegalovirus (CMV), Epstein Barr Virus (EBV), E. coli, Salmonella, Shigella, Staphylococcus aureus, Coliform Bacteria, Chlamydia, Mycobacterium Tuberculosis, Streptococcus, Pneumococcus, Pseudomonas, Campylobacter, Salmonella, Aspergillus Fumigatus, Aspergillus flavus, Cryptococcus Neoformans, and Histoplasma Capsulatum.