Methods and compositions for treatment of cancer with Anti-CD27 antibodies

Abstract

The present disclosure provides methods, compositions, uses, and kits for treatment of a cancer, e.g., a cancer having a PD-L1 CPS <10 (e.g., a TNBC having a PD-L1 CPS <10) using an anti-CD27 antibody or antigen-binding fragment thereof, alone or in combination with one or more additional therapeutic agents (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof and / or one or more chemotherapeutic agents).

Description

METHODS AND COMPOSITIONS FOR TREATMENT OF CANCER WITH ANT1-CD27ANTIBODIESCROSS-REERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 561,471, filed March 5, 2024, and U.S. Provisional Application No. 63 / 574,605, filed April 4, 2024, the disclosure of each of which is incorporated herein by reference in its entirety.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The sequence listing of the present application is submitted electronically as an XML formatted sequence listing created on July 31, 2024, with the file name “25916-WO-PCT_SL.xml” and having a size of 24,921 bytes. This sequence listing submitted electronically is part of the specification and is herein incorporated by reference in its entirety.FIELD

[0003] This disclosure relates generally to methods, compositions, uses, and kits for treatment of cancer (e.g., triple-negative breast cancer (TNBC)) using anti-CD27 antibodies.BACKGROUND

[0004] Cluster of differentiation (CD) 27, a tumor necrosis factor (TNF) receptor family super member, was identified as a membrane molecule on human T cells. According to current evidence, CD27 has a single ligand, CD70, which is a TNF family member. CD27 is currently thought to be exclusively expressed by hematopoietic cells, in particular those of the lymphocyte lineage, i.e., T lymphocyte (T cell), B-lymphocyte (B cell) and natural killer (NK) cells. CD27 was originally defined as a human T-cell co-stimulatory molecule that increments the proliferative response to T-Cell Receptor (TCR). The presence of CD70, the ligand of CD27, dictates the timing and persistence of CD27-mediated co-stimulation.

[0005] Transgenic expression of CD70 in immature dendritic cells was sufficient to convert immunological tolerance to virus or tumors into CD8+ T cell responsiveness. Likewise, agonistic soluble CD70 promoted the CD8+ T cell response upon such peptide immunization, and in CD70 transgenic mice, CD4+ and CD8+ effector cell formation in response to TCR stimulation was greatly facilitated. In mouse lymphoma models, tumor rejection was improved upon CD70 transgenesis or injection of an anti-mouse CD27 antibody.

[0006] Currently used chemotherapies for the treatment of metastatic TNBC lead to short-term responses in a minority of patients and have considerable toxicities.

[0007] Therefore, there is a need in the art for improved methods and compositions for treatment of cancer.SUMMARY

[0008] The present disclosure provides, inter alia, methods, compositions, uses, and kits for treatment of a cancer, e.g., a cancer having a PD-L1 CPS <10 (e.g., a triple-negative breast cancer having a PD-L1 CPS <10).

[0009] In one aspect, provided herein is a method for treating a cancer in a patient, the method comprising administering to the patient a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof, wherein a tumor sample obtained from the patient has been determined to have a programmed death-ligand 1 (PD-L1) combined positive score (CPS) of less than (<) 10. In some embodiments, the tumor sample is obtained from the patient prior to administration of the anti-CD27 antibody or the antigen-binding fragment thereof.

[0010] In another aspect, provided herein is a method for treating a cancer in a patient, the method comprising: a) determining that a tumor sample obtained from the patient has a PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof to the patient.

[0011] In another aspect, provided herein is an anti-CD27 antibody or an antigen-binding fragment thereof for use in treating a cancer in a patient, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of <10. In some embodiments, the tumor sample is obtained from the patient prior to administration of the anti-CD27 antibody or the antigen-binding fragment thereof.

[0012] In another aspect, provided herein is an anti-CD27 antibody or an antigen-binding fragment thereof for use in treating a cancer in a patient, the method comprising: a) determining that a tumor sample obtained from the patient has a PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof to the patient.

[0013] In another aspect, provided herein is a method for treating a previously untreated locally recurrent unresectable triple-negative breast cancer (TNBC) or metastatic TNBC in a patient, the method comprising administering to the patient a therapeutically effective amount of boserolimab, pembrolizumab, and nab-paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0014] In another aspect, provided herein is boserolimab for use in treating a previously untreated locally recurrent unresectable TNBC or metastatic TNBC in a patient, wherein the boserolimab is administered in combination with pembrolizumab and nab-paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0015] In another aspect, provided herein is a pharmaceutical composition for treating a previously untreated locally recunent unresectable TNBC or metastatic TNBC in a patient, the pharmaceutical composition comprising boserolimab, wherein the boserolimab is for use in combination with pembrolizumab and nab-paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0016] In another aspect, provided herein is a pharmaceutical composition for treating a previously untreated locally recurrent unresectable TNBC or metastatic TNBC in a patient, the pharmaceutical composition comprising pembrolizumab, wherein the pembrolizumab is for use in combination with boserolimab and nab-paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0017] In another aspect, provided herein is a pharmaceutical composition for treating a previously untreated locally recurrent unresectable TNBC or metastatic TNBC in a patient, the pharmaceutical composition comprising nab-paclitaxel, wherein the nab-paclitaxel is for use in combination with boserolimab and pembrolizumab. wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0018] In some embodiments of any of the preceding aspects, the cancer is triple-negative breast cancer (TNBC).

[0019] In some embodiments of any of the preceding aspects, the TNBC is (i) locally recurrent unresectable TNBC or (ii) metastatic TNBC. In some embodiments, the TNBC is locally recurrent unresectable TNBC. In other embodiments, the TNBC is metastatic TNBC.

[0020] In some embodiments of any of the preceding aspects, the patient is previously untreated for the cancer.

[0021] In some embodiments of any of the preceding aspects, the patient has not been previously treated with a chemotherapy.

[0022] In some embodiments of any of the preceding aspects, the patient has not been previously treated with an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or an anti- PD-L1 antibody).

[0023] In some embodiments of any of the preceding aspects, the PD-L1 CPS is determined using an immunohistochemistry (IHC) assay comprising the diagnostic anti-PD-Ll antibody

[0024] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or the antigen-binding fragment thereof comprises: a) a heavy chain variable region complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:2; c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:3; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:4; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:5; and I a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:6.

[0025] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or the antigen-binding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising SEQ ID NO:7 and the light chain comprises a light chain variable region comprising SEQ ID NO:9.

[0026] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or the antigen binding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 8 and the light chain comprises the amino acid sequence of SEQ ID NO: 10.

[0027] In some embodiments of any of the preceding aspects, the anti-CD27 antibody is boserolimab.

[0028] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion.

[0029] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 30 mg.

[0030] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient every three weeks (Q3W).

[0031] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient every six weeks (Q6W).

[0032] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 30 mg every three weeks (Q3W).

[0033] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 30 mg every six weeks (Q6W).

[0034] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is co-administered with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti-PD-Ll antibody or an antigen binding fragment thereof.

[0035] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is co-formulated with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti-PD-Ll antibody or an antigen binding fragment thereof.

[0036] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody, or the antigen binding fragment thereof comprises: a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 16; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 17; c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 18; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 11 ; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 12; and f) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 13.

[0037] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 14.

[0038] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody comprises a heavy chain and a light chain, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:20 and the light chain comprises the amino acid sequence of SEQ ID NO: 15.

[0039] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody is pembrolizumab.

[0040] In other embodiments of any of the preceding aspects, the anti-PD-1 antibody is a pembrolizumab variant.

[0041] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion.

[0042] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 400 mg.

[0043] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient every six weeks (Q6W).

[0044] In some embodiments of any of the preceding aspects, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 400 mg every six weeks (Q6W).

[0045] In some embodiments of any of the preceding aspects, the anti-CD27 antibody or antigen binding fragment thereof is co-administered with one or more chemotherapeutic agents.

[0046] In some embodiments of any of the preceding aspects, the one or more chemotherapeutic agents comprise nab-paclitaxel.

[0047] In some embodiments of any of the preceding aspects, the nab-paclitaxel is administered to the patient via intravenous infusion.

[0048] In some embodiments of any of the preceding aspects, the nab-paclitaxel is administered to the patient at a dose of about 100 mg / m2

[0049] In some embodiments of any of the preceding aspects, the nab-paclitaxel is administered to the patient at a dose of about 80 mg / m2.

[0050] In some embodiments of any of the preceding aspects, the nab-paclitaxel is administered to the patient at a dose of about 64 mg / m2.

[0051] In some embodiments of any of the preceding aspects, the nab-paclitaxel is administered to the patient on a 28-day cycle, and the nab-paclitaxel is administered to the patient on Day I. 8, and 15 of the 28-day cycle.

[0052] In some embodiments of any of the preceding aspects, the nab-paclitaxel is administered to the patient at a dose of about 100 mg / m2on a 28-day cycle, and the nab-paclitaxel is administered to the patient on Day 1, 8, and 15 of the 28-day cycle.

[0053] In some embodiments of any of the preceding aspects, the patient is greater than or equal to 18 years of age.

[0054] In another aspect, provided herein is a kit comprising an anti-CD27 antibody or an antigen-binding fragment thereof, and instructions to administer the anti-CD27 antibody or the antigen-binding fragment thereof to a patient having a cancer, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of <10. In some embodiments, the kit further comprises a diagnostic anti-PD-Ll antibody. In some embodiments, the diagnostic anti- PD-L1 antibody is 22C3. In some embodiments, the kit further comprises one or more reagents for an IHC assay. In some embodiments, the anti-CD27 antibody is boserolimab.

[0055] The summary of the technology described above is non-limiting and other features and advantages of the technology will be apparent from the following detailed description, and from the claims.BRIEF DESCRIPTION OF THE DRAWINGS

[0056] FIG. 1 is a schematic diagram showing the MK-5890-001 study design. ECOG PS,Eastern Cooperative Oncology Group performance status; IV, intravenous; Q6W, every76 weeks.aPatients received study treatment until disease progression, unacceptable adverse events (AEs). intercurrent illness that prevents further treatment, investigator’s decision to withdraw treatment, withdrawal of consent, pregnancy, or noncompliance with study requirements.bThe starting dose of nab-paclitaxel was 100 mg / m2. Doses could be reduced to 80 mg / m2and then 64 mg / m2based on the occurrence of DLTs using the modified toxicity probability interval design.cFollowing safety evaluation, an additional ~30 patients were treated with a tolerable dose of nab-paclitaxel (100 mg / m2(3 weeks on / 1 week off)) for further evaluation of safety and antitumor activity.

[0057] FIG. 2 is a graph showing association between baseline PD-L1 CPS status and change from baseline in tumor size.

[0058] FIGS. 3A and 3B are graphs showing association between baseline TcelliniGEP (FIG. 3A) and confirmed response and baseline TMB and confirmed response (FIG. 3B).DETAILED DESCRIPTION

[0059] The present disclosure is directed to. inter alia, methods, compositions, uses, and kits for treatment of cancer. Pembrolizumab plus chemotherapy is an approved therapy for patients with locally recurrent unresectable or metastatic TNBC whose tumors express PD-L1 (CPS >10). However, there is unmet need for effective treatment options in patients with PD-L1 CPS <10.

[0060] In particular, the present disclosure provides methods, compositions, uses, and kits for treatment of cancer having a PD-L1 CPS <10, e.g., TNBC having a PD-L1 CPS <10. The present disclosure is based, at least in part, on the discovery described herein that treatment that includes an anti-CD27 antibody (e.g., boserolimab) showed evidence of antitumor activity in patients whose tumors express PD-L1 at the cutoff of CPS <10. See, e.g., Example 2.Definitions

[0061] Listed below are definitions of various terms used herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

[0062] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry are those well-known and commonly employed in the art.

[0063] As used herein, the articles '‘a” and “an” refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means oneelement or more than one element. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0064] Reference to “or” indicates either or both possibilities unless the context clearly dictates one of the indicated possibilities. In some cases, “and / or” was employed to highlight either or both possibilities.

[0065] The term “about,” when modifying the quantity (e g., mg) of a substance or composition, or the value of a parameter characterizing a step in a method, or the like, refers to variation in the numerical quantity that can occur, for example, through ty pical measuring, handling and sampling procedures involved in the preparation, characterization and / or use of the substance or composition; through inadvertent error in these procedures; through differences in the manufacture, source, or purity of the ingredients employed to make or use the compositions or carry' out the procedures; and the like. In certain embodiments, “about” can mean a variation of ± 0.1%, ± 0.5%. ± 1%, ± 2%, ± 3%, ± 4%, ± 5%. ± 6%, ± 7%, ± 8%, ± 9% or ± 10%.

[0066] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of “from 50 mg to 500 mg” is inclusive of the endpoints, 50 mg and 500 mg, and all the intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value; they are sufficiently imprecise to include values approximating these ranges and / or values.

[0067] As used herein, the term “comprising” may include the embodiments “consisting of’ and “consisting essentially7of.” The terms “comprise(s),” “include(s),” “having,” “has,” “may,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that require the presence of the named ingredients / steps and permit the presence of other ingredients / steps. However, such description should be construed as also describing compositions or processes as “consisting of’ and “consisting essentially of’ the enumerated components.

[0068] “Consists essentially of,” and variations such as “consist essentially of or “consisting essentially of,” as used herein, indicate the inclusion of any recited elements or group of elements, and the optional inclusion of other elements, of similar or different nature than the recited elements, that do not materially change the basic or novel properties of the specified dosage regimen, method, or composition. As a non-limiting example, an anti-CD27 antibody that consists essentially of a recited amino acid sequence may also include one or more amino acids, including substitutions of one or more amino acid residues, which do not materially affect the properties of the binding compound.

[0069] “Administration” and “treatment.” as it applies to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, refers to contact of an exogenous pharmaceutical, therapeutic, diagnostic agent, or composition to the animal, human, subject, cell, tissue, organ, or biological fluid. “Treat” or “treating” a cancer (e.g., a cancer having a PD-L1 CPS < 10, e.g., a TNBC having a PD-L1 CPS < 10), as used herein, means to administer an anti-CD27 antibody (e g., boserolimab), or antigen-binding fragment, alone or in combination with one or more additional therapeutic agents (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof and / or one or more chemotherapeutic agents) to a subject having a cancer, or diagnosed with cancer, to achieve at least one positive therapeutic effect, such as for example, reduced number of cancer cells, reduced tumor size, reduced rate of cancer cell infiltration into peripheral organs, or reduced rate of tumor metastasis or tumor growth. “Treatment” may include one or more of the following: inducing / increasing an antitumor immune response, decreasing the number of one or more tumor markers, halting or delaying the growth of a tumor or blood cancer or progression of disease such as cancer, stabilization of disease, inhibiting the growth or survival of tumor cells, eliminating or reducing the size of one or more cancerous lesions or tumors, decreasing the level of one or more tumor markers, ameliorating or abrogating the clinical manifestations of disease, reducing the severity or duration of the clinical symptoms of disease such as cancer, prolonging the survival of a patient relative to the expected survival in a similar untreated patient, and inducing complete or partial remission of a cancerous condition or other disease.

[0070] Positive therapeutic effects in cancer can be measured in a number of w ays (See, e.g., W. A. Weber, J. Nucl. Med. 50: 1 S-10S (2009)). For example, with respect to tumor growth inhibition, according to NCI standards, a T / C ^42% is the minimum level of anti-tumor activity. A T / C < 10% is considered a high anti-tumor activity level, with T / C (%) = Median tumor volume of the treated / Median tumor volume of the control x 100. In some embodiments, the treatment achieved by a therapeutically effective amount is any of progression free survival (PFS), disease free survival (DFS) or overall survival (OS). PFS, also referred to as “Time to Tumor Progression” indicates the length of time during and after treatment that the cancer does not grow, and includes the amount of time patients have experienced a complete response or a partial response, as w ell as the amount of time patients have experienced stable disease. DFS refers to the length of time during and after treatment that the patient remains free of disease. OS refers to a prolongation in life expectancy as compared to naive or untreated individuals or patients. While an embodiment of the treatment methods, compositions and uses of the present invention may not be effective in achieving a positive therapeutic effect in every patient, it should do so in a statistically significant number of subjects as determined by any statistical testknown in the art such as the Student's t-test. the chi2-test. the U-test according to Mann and Whitney, the Kruskal -Wallis test (H-test), Jonckheere-Terpstra-test and the Wilcoxon-test.

[0071] The term ‘‘patient” (alternatively referred to as “subject” or “individual” interchangeably herein) refers to a mammal (e.g., human, non-human primate (e.g., cynomolgus monkey), rat, mouse, dog, cat, rabbit) capable of being treated with the methods and compositions of the invention, most preferably a human. In some embodiments, the patient is an adult patient. In other embodiments, the patient is a pediatric patient.

[0072] A “biomarker” is an objectively measured indicator, compound, or molecule that reflects the presence, or likely progression, or successful treatment of a particular condition. Biomarkers have long been used in drug development, and the discovery and validation of new efficacy biomarkers is expected to improve predictive disease models, reduce the time and cost associated with drug development, and increase the success rate of translating experimental drugs into clinical therapeutics. In addition, biomarkers are valuable in early detection of disease development, changes in disease status, and effectiveness of behavioral modifications and therapeutics in disease control. An exemplary biomarker as disclosed herein includes PD-L1 (e.g., PD-L1 CPS).

[0073] As used herein, the term “antibody” refers to any form of antibody that exhibits the desired biological activity. Thus, it is used in the broadest sense and specifically covers, but is not limited to, monoclonal antibodies (including full length monoclonal antibodies comprising two light chains and two heavy chains), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), humanized antibodies, fully human antibodies, and chimeric antibodies.

[0074] In general, the basic antibody structural unit comprises a tetramer. Each tetramer includes two identical pairs of polypeptide chains, each pair having one “light” (e.g.. about 25 kDa) and one “heavy” chain (e.g., about 50-70 kDa). The amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy -terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Typically, human light chains are classified as kappa and lambda light chains. Furthermore, human heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a “J” region of about 12 or more amino acids, with the heavy chain also including a “D” region of about 10 more amino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, N.Y. (1989). Antibodies and antigen-binding fragments of any of these classes or subclasses of antibodies may be utilized and administered in the present disclosure.

[0075] "Variable regions7’ or “V region” as used herein means the segment of IgG chains which is variable in sequence between different antibodies. The V region typically extends to Kabat residue 109 in the light chain and 113 in the heavy chain. The variable regions of each light / heavy chain pair form the antibody binding site. The “heavy chain variable region” or “heavy chain variable domain” is also referred to herein as “VH,” and the “light chain variable region” or “light chain variable domain” is also referred to herein as “VL.” Thus, in general, an intact antibody has two binding sites. Except in bifunctional or bispecific antibodies, the two binding sites are, in general, the same.

[0076] Typically, the variable domains of both the heavy and light chains comprise three hypervariable regions, also called complementarity determining regions (referred to here as “CDRs”), which are located within relatively conserved framework regions (FR). The CDRs are usually aligned by the framework regions, enabling binding to a specific epitope. In general, from N-terminal to C- terminal, both light and heavy chains variable domains comprise FR1, CDR1. FR2, CDR2, FR3, CDR3, and FR4. The assignment of amino acids to each domain is, generally, in accordance with the definitions of Sequences of Proteins of Immunological Interest, Kabat, et al.; National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ. No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32: 1-75; Kabat, et al., (1977) J. Biol. Chem. 252:6609- 6616; Chothia. et al.. (1987) J Mol. Biol. 196:901-917 or Chothia, et al, (1989) Nature 342:878- 883.

[0077] “Kabat,” as used herein, means an immunoglobulin alignment and numbering system pioneered by Elvin A. Kabat ((1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health. Bethesda, Md.).

[0078] As used herein, the term “framework” or “FR” residues refers to those variable domain residues other than the hypervariable region residues defined herein as CDR residues.

[0079] As used herein, the term “hypervariable region” refers to the amino acid residues of an antibody or antigen-binding fragment thereof that are responsible for antigen-binding. The hypervariable region comprises amino acid residues from a CDR (i.e., CDRL1, CDRL2 and CDRL3 in the light chain variable domain and CDRH1, CDRH2 and CDRH3 in the heavy chain variable domain). See Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (defining the CDR regions of an antibody by sequence); see also Chothia and Lesk (1987) J. Mol. Biol. 196: 901- 917 (defining the CDR regions of an antibody by structure).

[0080] “Monoclonal antibody,” “mAb,” or “Mab,” as used herein, refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population areidentical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, polyclonal antibody preparations typically include a multitude of different antibodies having different amino acid sequences in their variable domains, particularly their CDRs, which are often specific for different epitopes. The modifier “monoclonal7’ indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al. (1975) Nature 256: 495, or may be made by recombinant DNA methods (see. e.g., U.S. Pat. No. 4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597, for example. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0081] As used herein, unless otherwise indicated, “antibody fragment.” “antigen binding fragment thereof,” or “antigen-binding fragment" refer to antigen-binding fragments of antibodies, i.e., antibody fragments that retain the ability to bind specifically to the antigen (e.g., CD27) bound by the full-length antibody, e.g., fragments that retain one or more CDR (e.g., CDR1, CDR2, and CDR3) regions. Examples of antigen-binding fragments include, but are not limited to. Fab, Fab’, F(ab’)2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules, e.g., sc-Fv; and multispecific antibodies formed from antibody fragments.

[0082] A “Fab fragment” is comprised of one light chain and the CHI and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule. A Fab fragment can be the product of papain cleavage of an antibody. In other examples, a Fab fragment may be produced using recombinant antibody production.

[0083] An “Fc” region contains two heavy’ chain fragments comprising the CH2 and CH3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domains.

[0084] A “Fab’ fragment” contains one light chain and a portion or fragment of one heavy chain that contains the VH domain and the CHI domain and also the region between the CHI and CH2 domains, such that an interchain disulfide bond can be formed between the tw o heavy chains of two Fab’ fragments to form a F(ab’)2 fragment.

[0085] A “F(ab’)2 fragment” contains two light chains and two heavy chains containing a portion of the constant region between the CHI and CH2 domains, such that an interchain disulfide bond is formed between the two heavy chains. A F(ab’)2 fragment thus is composed oftwo Fab’ fragments that are held together by a disulfide bond between the two heavy chains. A F(ab’)2 fragment can be the product of pepsin cleavage of an antibody.

[0086] The “Fv region’’ comprises the variable regions from both the heavy and light chains but lacks the constant regions.

[0087] The term “single-chain Fv” or “scFv” antibody refers to an antibody fragment comprising the VH and VL domains of an antibody, in which these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen-binding. For a review of scFv. see Pluckthun (1994) The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315. See also International Patent Application Publication No. WO 88 / 01649 and U.S. Pat. Nos. 4,946, 778 and 5,260,203. The scFv may have an orientation in the N-terminal to C-terminal direction ofVH-linker-VL or VL-linker-VH.

[0088] A “bivalent antibody” comprises two antigen-binding sites. In some instances, the two binding sites have the same antigen specificities. However, bivalent antibodies may be bispecific (see below).

[0089] As used herein, the term “diabodies” refers to small antibody fragments with two antigen-binding sites, which fragments comprise a VH connected to a VL in the same polypeptide chain (VH-VL or VL-VH). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary' domains of another chain and create two antigen-binding sites. Diabodies are described more fully in, e.g.. EP 404,097; WO 93 / 11161; and Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448. For a review of engineered antibody variants generally see Holliger and Hudson (2005) Nat. Biotechnol. 23: 1126-1136.

[0090] An antibody that “specifically binds to” a specified target protein (e.g., CD27) is an antibody that exhibits preferential binding to that target as compared to other proteins, but this specificity does not require absolute binding specificity. An antibody is considered “specific” for its intended target if its binding is determinative of the presence of the target protein in a sample, e.g., without producing undesired results such as false positives. Antibodies, or binding fragments thereof, useful in the present invention will bind to the target protein with an affinity' that is at least two-fold greater, preferably at least ten times greater, more preferably at least 20- times greater, and most preferably at least 100-times greater than the affinity with non-target proteins. As used herein, an antibody is said to bind specifically to a polypeptide comprising a given amino acid sequence, e.g., the amino acid sequence of a mature human CD27 or humanPD-1 molecule, if it binds to polypeptides comprising that sequence but does not bind to proteins lacking that sequence.

[0091] As used herein, a “chimeric antibody” is an antibody having the variable domain from a first antibody and the constant domain from a second antibody, where the first and second antibodies are from different species. See. e.g., U.S. Pat. No. 4.816.567; and Morrison et al.. (1984) Proc. Natl. Acad. Sci. USA 81 : 6851-6855. Typically, the variable domains are obtained from an antibody from an experimental animal (the “parental antibody”), such as a rodent, and the constant domain sequences are obtained from human antibodies, so that the resulting chimeric antibody will be less likely to elicit an adverse immune response in a human subject than the parental (e.g., mouse) antibody.

[0092] “Humanized antibody” refers to forms of antibodies that contain sequences from nonhuman (e.g., murine) antibodies as well as human antibodies. Such antibodies contain minimal sequence derived from non-human immunoglobulin. In general, the humanized antibody will comprise substantially all of at least one, and typically two. variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc). typically that of a human immunoglobulin. The prefix “hum”, “hu” or “h” is added to antibody clone designations when necessary to distinguish humanized antibodies from parental rodent antibodies. The humanized forms of rodent antibodies will generally comprise the same CDR sequences of the parental rodent antibodies, although certain amino acid substitutions may be included to increase affinity, increase stability of the humanized antibody, or for other reasons.

[0093] As used herein, “germline sequence” refers to a sequence of unrearranged immunoglobulin DNA sequences. Any suitable source of unrearranged immunoglobulin sequences may be used. Human germline sequences may be obtained, for example, from JOINSOLVER germline databases on the website for the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the United States National Institutes of Health. Mouse germline sequences may be obtained, for example, as described in Giudicelli et al. (2005) Nucleic Acids Res. 33:D256-D261.

[0094] An “anti-CD27 antibody” useful in the any of the treatment methods, compositions, uses, and kits of the present disclosure include monoclonal antibodies, or antigen binding fragments thereof, which specifically bind to human CD27. Alternative names or synonyms for CD27 include S152, S152. LPFS2, T14, TNFRSF7, and Tp55. In any of the treatment methods,compositions and uses of the present disclosure in which a human individual is being treated, the anti-CD27 antibody or antigen binding fragment thereof is a CD27 agonist. Human CD27 amino acid sequences can be found in NCBI Locus No.: NP_001233. Human CD70 amino acid sequences can be found in NCBI Locus No.: NP_001243.1 (isoform 1) and NP_001317261.1 (isoform 2), respectively. An anti-CD27 antibody may be a human antibody, a humanized antibody or a chimeric antibody, and may include a human constant region. In some embodiments the human constant region is selected from the group consisting of IgGl, IgG2, IgG3 and IgG4 constant regions, and in preferred embodiments, the human constant region is an IgGl or IgG4 constant region. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab-SH, F(ab’)2, scFv and Fv fragments. Examples of anti- CD27 antibodies as used herein include, but are not limited to, boserolimab (Merck and Co., Inc.), varlilumab (also known as CDX-1127, Celldex Therapeutics, Inc ), BMS-986215 (Bristol- Myers Squibb Company), BNT313 / GEN1053 (BioNTech SE / Genmab A / S), and hCD27. 15. BNT313 / GEN1053 is an anti-CD27 antibody based on HexaBody technology that is engineered to form an antibody hexamer upon binding to CD27 on the cell membrane of T cells. Anti-CD27 antibodies are further described, e.g., in WO 2018 / 058022, WO 2021 / 087016, W02012 / 004367, W02015 / 016718, WO 2008 / 051424, WO 2011 / 130434, WO 2019 / 195452, WO 2023 / 031473, and CN 116262788.

[0095] An ‘'anti-PD-1 antibody” useful in the any of the treatment methods, compositions, uses, and kits of the present disclosure include monoclonal antibodies, or antigen binding fragments thereof, which specifically bind to human PD-1. Alternative names or synonyms for PD-1 (also known as programmed cell death protein 1) and its ligands include: PDCD1, PD1, CD279 and SLEB2 for PD-1; PDCD1L1, PDL1, B7H1. B7-4. CD274 and B7-H for PD-L1; and PDCD1L2. PDL2, B7-DC, Btdc and CD273 for PD-L2. In any of the treatment methods, compositions, uses, and kits of the present disclosure in which a human individual is being treated, the PD-1 antibody or antigen binding fragment thereof is a PD-1 antagonist that blocks binding of human PD-L1 to human PD-1, or blocks binding of both human PD-L1 and PD-L2 to human PD-1. Human PD-1 amino acid sequences can be found in NCBI Locus No.: NP_005009. Human PD-L1 and PD-L2 amino acid sequences can be found in NCBI Locus No.: NP_054862 and NP_079515, respectively. An anti-PD-1 antibody may be a human antibody, a humanized antibody or a chimeric antibody, and may include a human constant region. In some embodiments the human constant region is selected from the group consisting of IgGl, IgG2, IgG3 and IgG4 constant regions, and in preferred embodiments, the human constant region is an IgGl or IgG4 constantregion. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab’-SH, F(ab’)2, scFv and Fv fragments.

[0096] “PD-L1” or “PD-L2” expression means any detectable level of expression of the designated PD-L protein on the cell surface or of the designated PD-L mRNA within a cell or tissue, unless otherwise defined. PD-L protein expression may be detected with a diagnostic PD-L antibody in an immunohistochemistry (IHC) assay of a tumor tissue section or by flow cytometry. Alternatively, PD-L protein expression by tumor cells may be detected by position emission tomography (PET) imaging, using a binding agent (e.g.. antibody fragment, affibody and the like) that specifically binds to the desired PD-L target, e.g., PD-L1 or PD-L2. Techniques for detecting and measuring PD-L mRNA expression include reverse transcription polymerase chain reaction (RT-PCR), real-time quantitative RT-PCR, and RNA sequencing (RNAseq).

[0097] Several approaches have been described for quantifying PD-L1 protein expression in IHC assays of tumor tissue sections. See, e.g., Thompson et al., PNAS 101 (49): 17174-17179 (2004); Thompson et al.. Cancer Res. 66:3381-3385 (2006); Gadiot et al., Cancer 117:2192-2201 (2011); Taube et al., Sci. Transl. Med 4, 127ra37 (2012); and Topalian et al., New Eng. J. Med. 366 (26): 2443-2454 (2012).

[0098] One approach employs a simple binary endpoint of positive or negative for PD-L1 expression, with a positive result defined in terms of the percentage of tumor cells that exhibit histologic evidence of cell-surface membrane staining. A tumor tissue section is counted as positive for PD-L1 expression is at least 1%, and preferably 5% of total tumor cells.

[0099] In another approach, PD-L1 expression in the tumor tissue section is quantified in the tumor cells as well as in infiltrating immune cells, which predominantly comprise lymphocytes. The percentage of tumor cells and infiltrating immune cells that exhibit membrane staining are separately quantified as < 5%, 5 to 9%, and then in 10% increments up to 100%. For tumor cells, PD-L1 expression is counted as negative if the score is < 5% score and positive if the score is > 5%. PD-L1 expression in the immune infiltrate is reported as a semi-quantitative measurement called the adjusted inflammation score (AIS), which is determined by multiplying the percent of membrane staining cells by the intensify of the infiltrate, which is graded as none (0), mild (score of 1 , rare lymphocytes), moderate (score of 2, focal infiltration of tumor by lymphohistiocytic aggregates), or severe (score of 3, diffuse infiltration). A tumor tissue section is counted as positive for PD-L1 expression by immune infiltrates if the AIS is > 5.

[0100] A tissue section from a tumor that has been stained by IHC with a diagnostic PD- LI antibody may also be scored for PD-L1 protein expression by assessing PD-L1 expression in both the tumor cells and infiltrating immune cells in the tissue section using a scoring process.See WO 2014 / 165422. One PD-L1 scoring process comprises examining each tumor nest in the tissue section for staining and assigning to the tissue section one or both of a modified H score (MHS) and a modified proportion score (MPS). To assign the MHS, four separate percentages are estimated across all of the viable tumor cells and stained mononuclear inflammatory cells in all of the examined tumor nests: (a) cells that have no staining (intensity = 0), (b) weak staining (intensity =1+), (c) moderate staining (intensity =2+) and (d) strong staining (intensity =3+). A cell must have at least partial membrane staining to be included in the weak, moderate or strong staining percentages. The estimated percentages, the sum of which is 100%, are then input into the formula of 1 x (percent of weak staining cells) + 2 x (percent of moderate staining cells) + 3 x (percent of strong staining cells), and the result is assigned to the tissue section as the MHS. The MPS is assigned by estimating, across all of the viable tumor cells and stained mononuclear inflammatory cells in all of the examined tumor nests, the percentage of cells that have at least partial membrane staining of any intensity, and the resulting percentage is assigned to the tissue section as the MPS. In some embodiments, the tumor is designated as positive for PD-L1 expression if the MHS or the MPS is positive.

[0101] Another method for scoring / quantifying PD-L1 expression in a tumor is the “combined positive score’' or “CPS,” which refers to an algorithm for determining a PD-L1 expression score from a tumor sample of a patient. The CPS is useful in selecting patients for treatment with particular treatment regimens including methods of treatment comprising administration of an anti-PD-1 antibody in which expression of PD-L1 is associated with a higher response rate in a particular patient population relative to same patient population that does not express PD-L1. The CPS is determined by determining the number of viable PD-L1 positive tumor cells, the number of viable PD-L1 negative tumor cells, and the number of viable PD-L1 positive mononuclear inflammatory cells (MIC) in a tumor tissue from a patient having a tumor and calculating the CPS using the following formula:(# PD-L1 positive tumor cells) + (# PD-L1 positive MIC) x 100%(# PD-L1 positive tumor cells) + (PD-L1 negative tumor cells).

[0102] Yet another scoring method for PD-L1 expression is the “TPS” or “tumor proportion score,” which is the percentage of tumor cells expressing PD-L1 on the cell membrane. TPS ty pically includes the percentage of neoplastic cells expressing PD-L1 at any intensity (weak, moderate, or strong), which can be determining using an immunohistochemical assay using a diagnostic anti-human PD-L1 mAb, e.g., antibody 20C3 and antibody 22C3. Cells are considered to express PD-L1 if membrane staining is present, including cells with partial membrane staining.

[0103] The level of PD-L1 mRNA expression may be compared to the mRNA expression levels of one or more reference genes that are frequently used in quantitative RT-PCR, such as ubiquitin C.

[0104] In some embodiments, a level of PD-L1 expression (protein and / or mRNA) by malignant cells and / or by infiltrating immune cells within a tumor is determined to be “overexpressed” or “elevated” based on comparison with the level of PD-L1 expression (protein and / or mRNA) by an appropriate control. For example, a control PD-L1 protein or mRNA expression level may be the level quantified in nonmalignant cells of the same type or in a section from a matched normal tissue. In some preferred embodiments. PD-L1 expression in a tumor sample is determined to be elevated if PD-L1 protein (and / or PD-L1 mRNA) in the sample is at least 10%, 20%, or 30% greater than in the control.

[0105] In some embodiments, a level of PD-L1 expression (protein and / or mRNA) by malignant cells and / or by infiltrating immune cells within a tumor is determined to be “underexpressed” or “decreased” based on comparison with the level of PD-L1 expression (protein and / or mRNA) by an appropriate control. For example, a control PD-L1 protein or mRNA expression level may be the level quantified in nonmalignant cells of the same type or in a section from a matched normal tissue. In some preferred embodiments, PD-L1 expression in a tumor sample is determined to be decreased if PD-L1 protein (and / or PD-L1 mRNA) in the sample is at least 10%, 20%, or 30% lower than in the control.

[0106] “Tissue section” refers to a single part or piece of a tissue sample, e.g., a thin slice of tissue cut from a sample of a normal tissue or of a tumor.

[0107] “Tumor” as it applies to a subject diagnosed with, or suspected of having, a cancer refers to a malignant or potentially malignant neoplasm or tissue mass of any size and includes primary tumors and secondary neoplasms. A solid tumor is an abnormal growth or mass of tissue that usually does not contain cysts or liquid areas. Different types of solid tumors are named for the type of cells that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancers of the blood) generally do not form solid tumors (National Cancer Institute, Dictionary of Cancer Terms).

[0108] “Conservatively modified variants” or “conservative substitution” refers to substitutions of amino acids in a protein with other amino acids having similar characteristics (e.g., charge, side-chain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity, and the like), such that the changes can frequently be made without altering the biological activity of the protein. Those of skill in this art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g.,Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th Ed.)). In addition, substitutions of structurally or functionally similar amino acids are less likely to disrupt biological activity. Exemplary' conservative substitutions are set forth in Table 1.Table 1. Exemplary Conservative Amino Acid Substitutions

[0109] "‘Function-conservative variants,” as used herein, refers to antibodies or fragments thereof in which one or more amino acid residues have been changed without altering a desired property, such as an antigen affinity and / or specificity. Such variants include, but are not limited to, replacement of an amino acid with one having similar properties, such as the conservative amino acid substitutions of Table 1.

[0110] Typically, an antibody or antigen-binding fragment thereof as used herein is modified in some way retains at least 10% of its binding activity (when compared to the parental antibody) when that activity is expressed on a molar basis. Preferably, an antibody or antigen-binding fragment thereof of the invention retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the CD27 binding affinity as the parental antibody. It is also intended that an antibody or antigen-binding fragment thereof of the invention can include conservative or non-conservative amino acid substitutions (e.g., conservative variants or function-conserved variants of the antibody) that do not substantially alter its biologic activity.

[0111] The term “effector function” as used herein refers to one or more of antibody dependent dell mediated cytotoxic activity (ADCC), complement-dependent cytotoxic activity (CDC) mediated responses, Fc-mediated phagocytosis or antibody dependent cellular phagocytosis (ADCP) and antibody recycling via the FcRn receptor.

[0112] As used herein, the terms “N-glycan” and “glycoform” are used interchangeably and refer to an N-linked oligosaccharide, for example, one that is attached by an asparagine-N- acetylglucosamine linkage to an asparagine residue of a polypeptide. N-linked glycoproteins contain an N-acetylglucosamine residue linked to the amide nitrogen of an asparagine residue in the protein. The predominant sugars found on glycoproteins are glucose, galactose, mannose, fucose, N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc) and sialic acid (e.g., N-acetyl-neuraminic acid (NANA)). The processing of the sugar groups occurs co-translationally in the lumen of the ER and continues post-translationally in the Golgi apparatus for N-linked glycoproteins.

[0113] N-glycans have a common pentasaccharide core of MansGlcN Ac2 (“Man” refers to mannose; “Glc” refers to glucose; and "NAc" refers to N-acetyl; GlcNAc refers to N- acetylglucosamine). Usually, N-glycan structures are presented with the non-reducing end to the left and the reducing end to the right. The reducing end of the N-glycan is the end that is attached to the Asn residue comprising the glycosylation site on the protein. N-glycans differ with respect to the number of branches (antennae) comprising peripheral sugars (e.g., GlcNAc, galactose, fucose and sialic acid) that are added to the Man3GlcNAc2 (“Mans”) core structure which is also referred to as the “trimannose core,” the “pentasaccharide core,” or the “paucimannose core.” N- glycans are classified according to their branched constituents (e.g., high mannose, complex or hybrid). A “high mannose” type N-glycan has five or more mannose residues. A “complex” type N-glycan typically has at least one GlcNAc attached to the 1,3 mannose arm and at least one GlcNAc attached to the 1,6 mannose arm of a “trimannose” core. Complex N-glycans may also have galactose (“Gal”) or N- acetylgalactosamine (“GalNAc”) residues that are optionally modified with sialic acid or derivatives (e.g., “NANA” or “NeuAc.” where “Neu” refers to neuraminic acid and “Ac” refers to acetyl). Complex N-glycans may also have intrachain substitutions comprising “bisecting” GlcNAc and core fucose (“Fuc”). Complex N-glycans may also have multiple antennae on the “trimannose core,” often referred to as “multiple antennary glycans.” A “hybrid” N-glycan has at least one GlcNAc on the terminal of the 1,3 mannose arm of the trimannose core and zero or more mannoses on the 1 ,6 mannose arm of the trimannose core. The various N-glycans are also referred to as “glycoforms.”

[0114] "Isolated” antibodies or antigen-binding fragments thereof are at least partially free of other biological molecules from the cells or cell cultures in which they are produced. Such biological molecules include nucleic acids, proteins, lipids, carbohydrates, or other material such as cellular debris and growth medium. An isolated antibody or antigen-binding fragment thereof may further be at least partially free of expression system components such as biological molecules from a host cell or of the growth medium thereof. Generally, the term '‘isolated’’ is not intended to refer to a complete absence of such biological molecules or to an absence of water, buffers, or salts or to components of a pharmaceutical formulation that includes the antibodies or fragments.

[0115] “Isolated nucleic acid molecule” or “isolated polynucleotide” means a DNA or RNA of genomic, mRNA, cDNA, or synthetic origin or some combination thereof which is not associated with all or a portion of a polynucleotide in w hich the isolated polynucleotide is found in nature, or is linked to a polynucleotide to which it is not linked in nature. For purposes of this disclosure, it should be understood that “a nucleic acid molecule comprising” a particular nucleotide sequence does not encompass intact chromosomes. Isolated nucleic acid molecules comprising specified nucleic acid sequences may include, in addition to the specified sequences, coding sequences for up to ten or even up to twenty or more other proteins or portions or fragments thereof, or may include operably linked regulator} • sequences that control expression of the coding region of the recited nucleic acid sequences, and / or may include vector sequences.

[0116] The phrase “control sequences” refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism. The control sequences that are suitable for prokaryotes, for example, include a promoter, optionally an operator sequence, and a ribosome binding site. Eukaryotic cells are known to use promoters, polyadenylation signals, and enhancers.

[0117] “Percentage of sequence identity,” “percent sequence identity,” “percent identity,” and “percent identical” are used herein to refer to comparisons between polynucleotide sequences or polypeptide sequences, and are determined by comparing two optimally aligned sequences over a comparison window, in which the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence for optimal alignment of the tw o sequences. The percentage is calculated by determining the number of positions at which either the identical nucleic acid base or amino acid residue occurs in both sequences or a nucleic acid base or amino acid residue is aligned with a gap to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yieldthe percentage of sequence identity. Determination of optimal alignment and percent sequence identity can be performed using the BLAST and BLAST 2.0 algorithms (see e.g., Altschul et al., 1990, J. Mol. Biol. 215: 403-410; and Altschul et al., 1977, Nucleic Acids Res. 3389-3402). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information website.

[0118] Briefly, the BLAST analyses involve first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) of 10, M = 5, N = -4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word length (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff. 1989, PROC. NATL. ACAD. SCI. USA 89: 10915).

[0119] Numerous other algorithms are available that function similarly to BLAST in providing percent identity for two sequences. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith and Waterman, 1981. Adv. Appl. Math. 2:482, by the homology alignment algorithm of Needleman and Wunsch, 1970, J. Mol. Biol. 48:443, by the search for similarity method of Pearson and Lipman, 1988, Proc. Natl. Acad. Sci. USA 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the GCG Wisconsin Software Package), or by visual inspection (see generally, Current Protocols in Molecular Biolog}', F. M. Ausubel et al., eds., Current Protocols, ajoint venture between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (1995 Supplement)). Additionally, determination of sequence alignment and percent sequence identitycan employ the BESTF1T or GAP programs in the GCG Wisconsin Software package (Accelrys, Madison WI), using default parameters provided.

[0120] A nucleic acid or polynucleotide is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence. For example, DNA for a pre-sequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, but not always, “operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.

[0121] As used herein, the expressions “cell,” “cell line.” and “cell culture” are used interchangeably and all such designations include progeny. Thus, the words “transformants” and “transformed cells” include the primary subject cell and cultures derived therefrom without regard for the number of transfers. It is also understood that not all progeny will have precisely identical DNA content, due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where distinct designations are intended, it will be clear from the context.

[0122] The term “solid tumor cancer” is defined as a cancer disease wherein the malignant tumor is present at a distinct location and does not contain a cyst and / or liquid area. Solid tumors may be benign (not cancer), or malignant (cancer). Bladder cancer, breast cancer (e.g.. TNBC), colon and rectal cancer (colorectal cancer), endometrial cancer, kidney (renal cell) cancer, lung cancer, pancreatic cancer, prostate cancer, and thyroid cancer are non-limiting examples of different types of solid tumor cancers. Leukemia is an example of a cancer that is not a solid tumor cancer. See U.S. Patent No. 10,451.626.

[0123] The term “triple-negative breast cancer” as used herein refers to a breast cancer that tests negative for estrogen receptors, progesterone receptors, and HER2.

[0124] The term “monotherapy” refers to the use of a single agent or one type of treatment (e.g., radiation therapy) to treat a certain disease or conditions, e.g.. a cancer.

[0125] The term “in association with” indicates that the components administered to a subject, e.g., an anti-CD27 antibody or antigen-binding fragment thereof along w ith one or more additional therapeutic agents can be formulated into a single composition for simultaneousdelivery or formulated separately into two or more compositions (e.g., a kit). Each component can be administered to a subject at a different time than when the other component is administered; for example, each administration may be given non-simultaneously (e.g., separately or sequentially) at several intervals over a given period of time. Moreover, the separate components may be administered to a subject by the same or by a different route. The term “in association with’’ encompasses co-administration and co-formulation.

[0126] “Co-administration" as used herein for therapeutic agents (e.g., an anti-CD27 antibody or antigen binding fragment thereof, and one or more additional therapeutic agents, such as an anti-PD-1 antibody or antigen binding fragment thereof and / or one or more chemotherapeutic agents) means that the agents are administered so as to have overlapping therapeutic activities, and not necessarily that the agents are administered simultaneously to the subject. The agents may or may not be in physical combination prior to administration. In an embodiment, the agents are administered to a subject simultaneously or at about the same time. For example, an anti-PD- 1 antibody and an anti-CD27 antibody contained in separate vials, when in liquid solution, may be mixed into the same intravenous infusion bag or injection device, and administered simultaneously to the patient. In other embodiments, the agents are administered sequentially. For example, an anti-CD27 antibody or antigen-binding fragment thereof may be administered prior to or following one or more additional therapeutic agents, e.g., an anti-PD-1 antibody or antigen binding fragment thereof and / or one or more chemotherapeutic agents.

[0127] “Co-formulated” or “co-formulation” as used herein refers to at least two agents (e.g., different antibodies or antigen binding fragments) thereof which are formulated together and stored as a combined product in a single vial or vessel (e.g.. an injection device) rather than being formulated and stored individually and then mixed before administration or separately administered. In one embodiment, the co-formulation contains two different antibodies or antigen binding fragments thereof described herein, for example, an anti-CD27 antibody (e.g., boserolimab) or antigen binding fragment thereof described herein and an anti-PD-1 antibody (e.g.. pembrolizumab) or antigen binding fragment described herein.

[0128] “Chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Classes of chemotherapeutic agents include, but are not limited to: platinum-containing chemotherapeutic agents (e.g., cisplatin, carboplatin, and oxaliplatin), alkylating agents, antimetabolites, kinase inhibitors, spindle poison plant alkaloids, cytotoxic / cytostatic agents, topoisomerase inhibitors, photosensitizers, anti-estrogens and selective estrogen receptor modulators (SERMs), anti -progesterones, estrogen receptor down-regulators (ERDs), estrogen receptor antagonists, luteinizing hormone-releasing hormone agonists, anti-androgens, aromataseinhibitors, EGFR inhibitors. VEGF inhibitors, anti-sense oligonucleotides that that inhibit expression of genes implicated in abnormal cell proliferation or tumor growth.

[0129] “Cytotoxic / cytostatic agents’’ refer to compounds which cause cell death or inhibit cell proliferation primarily by interfering directly with the cell’s functioning or inhibit or interfere with cell myosis. including alkylating agents, tumor necrosis factors, intercal ators, hypoxia activatable compounds, microtubule inhibitors / microtubule-stabilizing agents, inhibitors of mitotic kinesins, histone deacetylase inhibitors, inhibitors of kinases involved in mitotic progression, inhibitors of kinases involved in growth factor and cytokine signal transduction pathways, antimetabolites, biological response modifiers, hormonal / anti-hormonal therapeutic agents, hematopoietic growth factors, monoclonal antibody targeted therapeutic agents, topoisomerase inhibitors, proteosome inhibitors, ubiquitin ligase inhibitors, and aurora kinase inhibitors.

[0130] As used herein, “therapeutically effective amount" refers to an amount of a therapeutic agent (e.g.. an anti-CD27 antibody (e.g.. boserolimab) or antigen binding fragment thereof), which is effective as a monotherapy or in combination with one or more active ingredients (e.g., an anti-PD-1 antibody or antigen binding fragment thereof, one or more chemotherapeutic agents, or a combination thereof), upon single or multiple dose administration (e.g., an infusion treatment over a period of time, such as 30 minutes, or a bolus and / or maintenance doses) to a subject, in inhibiting, delaying or reducing the growth of a cancer (e.g., a cancer characterized by presence of a solid tumor (e.g., a cancer having a PD-L1 CPS <10 (e.g., a TNBC having a PD-L1 CPS <10))). “Therapeutically effective” is also intended to refer to an amount of the therapeutic agent that is suitable for therapeutic uses. In various embodiments, a therapeutically effective amount of an anti-CD27 antibody or antigen binding fragment thereof is the amount of the anti- CD27 antibody or antigen binding fragment thereof that is needed to treat a solid tumor cancer in combination with one or more additional therapeutic agents, for example an anti-PD-1 antibody such as pembrolizumab, one or more chemotherapeutic agents, or a combination thereof.

[0131] A therapeutically effective dose of any of the antibodies or antigen binding fragments thereof described herein (e.g., in Table 3 or Table 4), administered alone or in combination with one or more additional therapeutic agents to a cell, tissue, or subject, is effective to cause a measurable improvement in one or more symptoms of disease (e.g., cancer) or the progression of the disease. A therapeutically effective dose may be an amount of the antibody or fragment thereof sufficient to result in at least partial amelioration of symptoms, e.g., tumor shrinkage or elimination, lack of tumor growth, or increased survival time. When applied to an individual active ingredient administered alone, an effective dose refers to that ingredient alone. Whenapplied to a combination, a therapeutically effective dose may be combined with amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. An effective amount of a therapeutic may result in an improvement of a biomarker, measure, or parameter by at least 10%; usually by at least 20%; preferably at least about 30%; more preferably at least 40%, and most preferably by at least 50%. An effective amount can also result in an improvement in a subjective measure in cases where subjective measures are used to assess disease severity.

[0132] ‘‘Diagnostic anti-PD-L monoclonal antibody” means a monoclonal antibody (mAb) which specifically binds to the mature form of the designated PD-L (PD-L1 or PD-L2) that is expressed on the surface of certain mammalian cells. A mature PD-L lacks the presecretory leader sequence, also referred to as leader peptide The terms “PD-L” and “mature PD-L” are used interchangeably herein, and shall be understood to mean the same molecule unless otherwise indicated or readily apparent from the context.

[0133] As used herein, a diagnostic anti-human PD-L1 mAb or an anti-hPD-L l mAb refers to a monoclonal antibody that specifically binds to mature human PD-L1. A mature human PD-L1 molecule consists of amino acids 19-290 of the following sequence:MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWE MEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMIS YGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLS GKTTTTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNER THLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET (SEQ ID NO: 21).

[0134] Specific examples of diagnostic anti-human PD-L1 mAbs useful as diagnostic mAbs for immunohistochemistry (IHC) detection of PD-L 1 expression in formalin-fixed, paraffin- embedded (FFPE) tumor tissue sections are antibody 20C3 and antibody 22C3, which are described in WO 2014 / 100079. These antibodies comprise the light chain and heavy chain variable region amino acid sequences shown in Table 2 below:Table 2. Monoclonal Antibodies 20C3 and 22C3

[0135] Another anti -human PD-L1 mAh that has been reported to be useful for IHC detection of PD-L1 expression in FFPE tissue sections (Chen, B.J. et al., Clin Cancer Res 19:3462-3473 (2013)) is a rabbit anti-human PD-L1 mAb publicly available from Sino Biological, Inc. (Beijing, P.R. China; Catalog number 10084-R015).Therapeutic Methods, Uses, and Compositions for Use

[0136] Provided herein are methods, compositions, and uses for treating patients, including human patients, in need of treatment with the antibodies or antigen-binding fragments thereof disclosed herein (e.g., an anti-CD27 antibody (e.g., boserolimab) or antigen-binding fragments thereof).

[0137] In one aspect, provided herein is a method for treating a cancer (e.g., TNBC) in a patient, the method comprising administering to the patient a therapeutically effective amount of an anti-CD27 antibody (e.g.. boserolimab) or an antigen-binding fragment thereof, wherein a tumor sample obtained from the patient has been determined to have a programmed death-ligand 1 (PD-L1) combined positive score (CPS) of less than (<) 10.

[0138] In another aspect, provided herein is a method for treating a cancer (e.g., TNBC) in a patient, the method comprising: a) determining that a tumor sample obtained from the patient hasa PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof to the patient.

[0139] In another aspect, provided herein is an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof for use in treatment of a cancer (e.g., TNBC) in a patient, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0140] In another aspect, provided herein is an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof for use in a method of treating a cancer (e.g., TNBC) in a patient, the method comprising: a) determining that a tumor sample obtained from the patient has a PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof to the patient.

[0141] In another aspect, provided herein is the use of an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof for treatment of a cancer (e.g., TNBC) in a patient, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0142] In another aspect, provided herein is the use of an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof in the manufacture of a medicament for treatment of a cancer (e.g.. TNBC) in a patient, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0143] In another aspect, provided herein is the use of an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof in a method of treating a cancer (e.g., TNBC) in a patient, the method comprising: a) determining that a tumor sample obtained from the patient has a PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof to the patient.

[0144] In another aspect, provided herein is the use of an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof in the manufacture of a medicament for treatment of a cancer (e.g.. TNBC) in a patient, the treatment comprising: a) determining that a tumor sample obtained from the patient has a PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof to the patient.

[0145] The method may be used in any suitable line of treatment. In some examples, the patient is previously untreated for the cancer (e.g., TNBC). In other words, the treatment may be a first- line (IL) therapy. In some examples, the patient has not been previously treated with achemotherapy. In some examples, the patient has not been previously treated with an immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-Ll antibody).

[0146] Any suitable approach may be used to determine the PD-L1 CPS. In some examples, the PD-L1 CPS is determined using an immunohistochemistry (IHC) assay, an immunofluorescence assay, or flow cytometry. Any suitable diagnostic anti-PD-Ll antibody (e.g., a diagnostic anti- PD-Ll monoclonal antibody) may be used, e g., 22C3 or 20C3. In some examples, the PD-L1 CPS is determined using an immunohistochemistry (IHC) assay comprising the diagnostic anti- PD-Ll antibody 22C3.

[0147] In some embodiments, the patient’s tumor has a decreased expression level of PD-L1 compared to a reference tumor.

[0148] In some embodiments, the patient’s tumor does not express PD-L1.

[0149] In some embodiments, the patient’s tumor has an expression level of PD-L1 that is equivalent to a CPS < 10, e.g., as assessed using a different diagnostic scoring approach (e.g., AIS, a modified H score (MHS) a modified proportion score (MPS), or a tumor proportion score (TPS).

[0150] Any suitable anti-CD27 antibody may be used, e.g., any anti-CD27 antibody or antigen binding fragment thereof described herein. See, e.g., the sub-section “Anti-CD27 Antibodies and Antigen-binding Fragments” below.

[0151] In some examples, the anti-CD27 antibody or the antigen-binding fragment thereof comprises one or more (e.g., 1, 2, 3, 4, 5, or all 6) of the following: a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:2; c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:3; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:4; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:5; and / or f) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:6.

[0152] In particular examples, the anti-CD27 antibody or the antigen-binding fragment thereof comprises: a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:1; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:2; c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:4; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:5; and f) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:6.

[0153] In some examples, the anti-CD27 antibody or the antigen-binding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising SEQ ID NO:7 and the light chain comprises a light chain variable region comprising SEQ ID NO:9.

[0154] In some examples, the anti-CD27 antibody or the antigen binding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 8 and the light chain comprises the amino acid sequence of SEQ ID NO: 10.

[0155] In some examples, the anti-CD27 antibody is boserolimab.

[0156] In some examples, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion.

[0157] In various embodiments, the antibody or antigen binding fragment thereof (e.g., an anti- CD27 antibody or antigen-binding fragment thereof, an anti-PD- 1 antibody or antigen-binding fragment thereof, or both) is administered at one dose. In various embodiments, the antibody or antigen binding fragment thereof is administered at multiple doses.

[0158] Any suitable dose or dosing regimen of the anti-CD27 antibody (e.g., boserolimab) or antigen binding fragment thereof may be used. For example, any dose or dosing regimen described in WO 2021 / 087016, which is incorporated by reference herein in its entirety, may be used.

[0159] In various embodiments, the anti-CD27 antibody (e.g., boserolimab) or antigen binding fragment thereof is administered a dose of about 2 mg to about 700 mg. For example, the subject or patient is administered about 2 mg, about 7 mg, about 20 mg, about 30 mg, about 70 mg, about 200 mg, or about 700 mg. In various embodiments of the method, the patient is administered 2 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered 7 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered 20 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered 30 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered 70 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered 200 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered 700 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered a dose of about 200 mg to about 700 mg of the anti-CD27 antibody or antigenbinding fragment thereof. In various embodiments of the method, the patient is administered a dose of about 30 mg to about 700 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered a dose of about 30 mg, about 200 mg, or about 200 mg to about 700 mg of the anti-CD27 antibody or antigen binding fragment. In various embodiments of the method, the patient is administered a dose of about 30 mg to about 200 mg of the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments of the method, the patient is administered a dose of about 70 mg to about 200 mg of the anti-CD27 antibody or antigen binding fragment thereof.

[0160] In some examples, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient even7three weeks (Q3W).

[0161] In other examples, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient every six weeks (Q6W).

[0162] In some examples, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 30 mg. In some examples, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient every six weeks (Q6W). In some examples, the anti-CD27 antibody (e.g., boserolimab) or antigen binding fragment thereof is administered to the patient at a dose of 30 mg every six weeks (Q6W). In other examples, the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient every three weeks (Q6W). In some examples, the anti-CD27 antibody (e.g., boserolimab) or antigen binding fragment thereof is administered to the patient at a dose of 30 mg every three weeks (Q3W).

[0163] In various embodiments, the anti-PD-1 antibody or antigen binding fragment thereof is administered as an infusion. In various embodiments, the anti-CD27 antibody or antigen binding fragment thereof is administered as an infusion over a period of time. For example, the period of time is at least about 30 minutes. In various embodiments, the period of time is 30 minutes.

[0164] In some examples, the anti-CD27 antibody or antigen binding fragment thereof is coadministered with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti- PD-L1 antibody or an antigen binding fragment thereof. In some examples, the anti-CD27 antibody or antigen binding fragment thereof is co-formulated with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti-PD-Ll antibody or an antigen binding fragment thereof.

[0165] In various embodiments, the anti-CD27 antibody or antigen binding fragment thereof is administered from a vial. For example, the volume of the vial is about 1 milliliter (mL or ml).

[0166] In various embodiments, the concentration of the anti-CD27 antibody or antigen binding fragment thereof in the vial is about 50 mg / ml.

[0167] In various embodiments, the intravenous infusion comprises administering an analgesic and / or an antihistamine prior to the anti-CD27 antibody or antigen binding fragment thereof. In various embodiments, the antihistamine is diphenhydramine. In various embodiments, the analgesic and / or antihistamine is administered less than 3 hours, 2.5 hours, 2 hours, 1.5 hours, or 1 hour prior to administration of the anti-CD27 antibody or antigen binding fragment thereof. For example, diphenhydramine is administered at a dose of about 50 mg. For example, diphenhydramine is administered orally. In various embodiments, the analgesic is acetaminophen. For example, acetaminophen is administered at a dose of about 500 to about 1000 mg. In various embodiments, acetaminophen is administered orally.

[0168] Any suitable anti-PD-1 antibody or antigen binding fragment thereof may be used, e.g., any anti-PD-1 antibody or antigen binding fragment thereof described herein. See, e.g., the subsection “Anti-PD-1 Antibodies and Antigen-binding Fragments” below.

[0169] In some examples, the anti-PD-1 antibody, or the antigen binding fragment thereof comprises: a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 16; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:17; c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 18; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 11: e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 12; and f) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:13.

[0170] In some examples, the anti-PD-1 antibody comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 14.

[0171] In some examples, the anti-PD-1 antibody comprises a heavy chain and a light chain, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:20 and the light chain comprises the amino acid sequence of SEQ ID NO: 15.

[0172] In particular examples, the anti-PD-1 antibody is pembrolizumab. In other examples, the anti-PD-1 antibody is a pembrolizumab variant.

[0173] Any suitable administration route may be used for the anti-PD-1 antibody or antigenbinding fragment thereof. In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient intravenously. In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion (e.g.,over 30 minutes). In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient subcutaneously.

[0174] Any suitable dose of the anti-PD-1 antibody or antigen binding fragment thereof may be used. In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 200 mg. In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 200 mg every' three weeks (Q3W). In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 400 mg. In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient every’ six weeks (Q6W). In some examples, the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 400 mg every six weeks (Q6W). In other examples, the anti-PD-1 antibody or antigen binding fragment thereof may be administered at a dose of 2 mg / kg, e.g., 2 mg / kg up to 200 mg for pediatric patients. In other examples, the anti-PD-1 antibody or antigen binding fragment thereof may be administered at a dose of 2 mg / kg every three weeks (Q3W), e.g., 2 mg / kg up to 200 mg Q3W for pediatric patients.

[0175] In various embodiments, the anti-PD-1 antibody or antigen binding fragment thereof is administered as an infusion. In various embodiments, the anti-CD27 antibody or antigen binding fragment thereof is administered as an infusion over a period of time. For example, the period of time is at least about 30 minutes. In various embodiments, the period of time is 30 minutes.

[0176] In some examples, the patient is greater than or equal to 18 years of age. In some examples, the patient is an adult. In other examples, the patient is less than 18 years of age, e.g., a pediatric patient.

[0177] In various embodiments, the subject has not been previously treated with anti-PD-1 or anti-PD-Ll therapy or is confirmed progressive while receiving prior anti-PD-1 or anti-PD-Ll therapy.

[0178] Any suitable cancer may be treated using the methods, compositions, uses, and kits disclosed herein. In various embodiments, the anti-CD27 antibody or antigen binding fragment thereof is used to treat a patient having a solid tumor and / or a cancer characterized by presence of a solid tumor. A solid tumor in various embodiments is an abnormal mass of tissue. Solid tumors may be benign, or malignant. Different types of solid tumors are named for the ty pe of cells that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancers of the blood) generally do not form solid tumors. In various embodiments the solid tumor does not contain a cyst and / or a liquid area. Exemplary’ solid tumors include but are not limited to sarcomas and carcinomas such as fibrosarcoma, myxosarcoma, liposarcoma,chondrosarcoma. osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma. lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing’s tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer (e.g., triple-negative breast cancer (TNBC), lung cancer (e.g.. non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC)), ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma. Wilm’s tumor, cervical cancer, testicular tumor, bladder carcinoma, epithelial carcinoma, glioblastoma multiforme, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, melanoma, neuroblastoma, and retinoblastoma.

[0179] In one embodiment, cancers that may be treated by the antibodies or antigen-binding fragments thereof disclosed herein include, but are not limited to: breast cancer (e.g.. TNBC), lung cancer (e.g., NSCLC (including squamous NSCLC or non-squamous NSCLC) or SCLC), pancreatic cancer, colon cancer, colorectal cancer, myeloid leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, chronic myelomonocytic leukemia, thyroid cancer, myelodysplastic syndrome, bladder carcinoma, epidermal carcinoma, melanoma, prostate cancer, head and neck cancers, ovarian cancer, brain cancers, cancers of mesenchymal origin (e.g., sarcomas), neuroblastomas, kidney carcinomas, hepatomas, non-Hodgkin’s lymphoma, multiple myeloma, and anaplastic thyroid carcinoma. In some examples, the cancer is a metastatic cancer, e.g., of the varieties described above. For example, the cancer may be a metastatic breast cancer (e.g.. metastatic TNBC).

[0180] In some examples of any of the preceding aspects, the cancer is TNBC. In some examples, the TNBC is (i) locally recurrent unresectable TNBC or (ii) metastatic TNBC. In some examples, the TNBC is locally recurrent unresectable TNBC. In other examples, the TNBC is metastatic TNBC.

[0181] In other examples, the cancer is selected from the group consisting of: TNBC, NSCLC, small cell lung cancer (SCLC), and endometrial cancer. In some examples, the cancer is NSCLC. In other examples, the cancer is SCLC. In other examples, the cancer is endometrial cancer.

[0182] In some examples, the patient is non-TcellinfGEP-high (e.g.. less than -0.318). In other examples, the patient is TcellinfGEP-high (e.g., greater than or equal to -0.318).

[0183] In some examples, the patient’s tumor is TMB-low (e.g., <10 mut / Mb, e.g., 1 mut / Mb, 2 mut / Mb, 3 mut / Mb, 4 mut / Mb, 5 mut / Mb, 6 mut / Mb, 7 mut / Mb, 8 mut / Mb, or 9 mut / Mb). Inother examples, the patient’s tumor is TMB-high (e.g., greater than or equal to 10 mut / Mb, e.g., 10 mut / Mb, 11 mut / Mb, 12 mut / Mb, 13 mut / Mb, 14 mut / Mb, 15 mut / Mb, 16 mut / Mb, 17 mut / Mb, 18 mut / Mb, 19 mut / Mb, 20 mut / Mb, or higher).

[0184] In particular embodiments, the antibodies or antigen-binding fragments thereof disclosed herein may be used alone, or in association with other, additional therapeutic agents and / or therapeutic procedures, for treating or preventing any disease such as cancer, e g., as discussed herein, in a subject in need of such treatment or prevention. Compositions, e.g., pharmaceutical compositions comprising a pharmaceutically acceptable carrier, comprising such antibodies and fragments thereof in association with further therapeutic agents are also part of the present disclosure.

[0185] Therefore, the present disclosure provides a method or use of treating cancer in a subject (e.g., a human subject), comprising administering to the subject an effective amount of an antibody or antigen-binding fragment thereof (e.g., an anti-CD27 antibody or antigen-binding fragment thereof, an anti-PD-1 antibody or antigen-binding fragment thereof, or both) disclosed herein, optionally in association with an additional therapeutic agent or therapeutic procedure. The present disclosure also provides a method of treating cancer in a human subject, comprising administering to the subject an effective amount of an antibody or antigen binding fragment thereof disclosed herein, optionally in association with an additional therapeutic agent or therapeutic procedure. The present disclosure also provides a method of increasing the activity of an immune cell, comprising administering to a subj ect in need thereof an effective amount of an antibody or antigen binding fragment thereof disclosed herein. In another embodiment, the present disclosure provides an antibody or antigen binding fragment thereof of the invention, for use in treatment of cancer as a monotherapy; increasing the activity of an immune cell; or treatment of cancer in combination with one or more additional therapeutic agents. In a further embodiment, the present disclosure provides use of the antibody or antigen binding fragment thereof of the invention for the manufacture of a medicament for increasing immune cell activation; treating cancer as a monotherapy; or treating cancer in combination with one or more additional therapeutic agents. In another embodiment, the present disclosure provides a combination of an antibody or antigen binding fragment thereof of the invention and a further therapeutic agent for the treatment of cancer; increasing the activity of an immune cell; or treatment of another disorder.

[0186] In other embodiments, the disclosure provides a method of treating cancer in a human subject, comprising administering to the subject an effective amount of an antibody or antigen binding fragment thereof disclosed herein, or an expression vector or a host cell according to thedisclosure. optionally in association with one or more additional therapeutic agents and / or therapeutic procedures.

[0187] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein (e.g., an anti-CD27 antibody or antigen-binding fragment thereof, an anti-PD-1 antibody or antigen-binding fragment thereof, or both) may be used in association with a tumor vaccine. Examples of tumor vaccines include but are not limited to vaccines for Human Papillomavirus (HPV) infection-caused cancer such as Gardasil®, Gardasil® and Cervarix®; vaccines that prevent hepatitis B virus-caused liver cancer such as Engerix-B® and Recombivax HB®; oncolytic virus therapy that triggers immune response such as Imlygic®; DNA vaccines such as Synchotrope MA2M plasmid DNA vaccine and ZYC101; mammaglobin-a DNA vaccine (see Clinical Cancer Res. 2014 20(23):5964-75); vector based vaccines such as PSA-TRICOM (prostvac), PANVAC-VF, Listeria monocytogenes -based PSA vaccine (see Therapeutic Advances in Vaccines. 2014, 2(5) 137- 148), ZAtena-mesothelin Adeno-CEA; allogeneic vaccines such as GV AX. BLP-25 (anti-Ankara-mucin 1). Belagenpumatucel-L, TG4010, CIMAvax epidermal grow th factor vaccine, NY-ESO, GM.CD40L-CCL21; autologous vaccines such as:Adeno-CD40L, BCG, INGN-225, dendritic cell vaccines such as Provenge® (Sipuleucel- T), rF-CEA-MUCl-TRICOM (panvac-DC); antigen vaccines such as MUC-1 (stimuvax), NY- ESO- 1. GP-100, MAGE-A3 (melanoma antigen encoding gene A3), or INGN-225 (see Pharmacology & Therapeutics 153 (2015) 1-9).

[0188] In particular embodiments, the anti-CD27 antibodies or antigen- binding fragments thereof disclosed herein may be used in association with radiation therapy.

[0189] In particular embodiments, the anti-CD27 antibodies or antigen-binding fragments thereof disclosed herein may be used in association with targeted therapies. Examples of targeted therapies include: hormone therapies, signal transduction inhibitors (e g., EGFR inhibitors, such as cetuximab (Erbitux®) and erlotinib (Tarceva®)); HER2 inhibitors (e.g., trastuzumab (Herceptin®) and pertuzumab (Perjeta®)); BCR-ABL inhibitors (such as imatinib (Gleevec®) and dasatinib (Sprycel®)); ALK inhibitors (such as crizotinib (Xalkori®) and ceritinib (Zykadia®)); BRAF inhibitors (such as vemurafenib (Zelboraf®) and dabrafenib (Tafinlar®)), gene expression modulators, apoptosis inducers (e.g., bortezomib (Velcade®) and carfilzomib (Kyprolis®)), angiogenesis inhibitors, e.g., bevacizumab (Avastin®) and ramucirumab (Cyramza®). monoclonal antibodies attached to toxins (e g., brentuximab vedotin (Adcetris®) and ado-trastuzumab emtansine (Kadcyla®)).

[0190] In particular embodiments, the anti-CD27 antibodies or antigen-binding fragments thereof may be used in association with an anti-cancer therapeutic agent or immunomodulatory'drug such as an immunomodulatory receptor inhibitor, e.g.. an antibody or antigen-binding fragment thereof that specifically binds to the receptor.

[0191] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with one or more of: an anti-PD-1 antibody (e.g., pembrolizumab (KEYTRUDA®). nivolumab. pidilizumab (CT-011)), an anti-PD-Ll antibody (e g., BMS-936559, Durvalumab, MSB0010718C or MPDL3280A), an anti-PD-L2 antibody, anti-TIGIT antibody, an anti-CTLA4 antibody, an anti-CSl antibody (e.g., elotuzumab), an anti- KIR2DL1 / 2 / 3 antibody (e.g., lirilumab), an anti-CD137 antibody (e.g., urelumab), an anti-GITR antibody (e.g., TRX518), an anti-ILTl antibody, an anti-ILT2 antibody, an anti-ILT3 antibody, an anti-ILT4 antibody, an anti-ILT5 antibody, an anti-ILT6 antibody, an anti-ILT7 antibody, an anti-ILT8 antibody, an anti-CD40 antibody, an anti-OX40 antibody, an anti-ICOS, an anti- SIRPa, an anti-KIR2DLl antibody, an anti-KIR2DL2 / 3 antibody, an anti-KIR2DL4 antibody, an anti-KIR2DL5A antibody, an anti-KIR2DL5B antibody, an anti-KIR3DLl antibody, an anti- KIR3DL2 antibody, an anti-KIR3DL3 antibody, an anti-NKG2A antibody, an anti-NKG2C antibody, an anti-NKG2E antibody, an anti-4- IBB antibody (e.g., PF-05082566), an anti-TSLP antibody, an anti-IL-10 antibody, IL-10 or PEGylated IL-10, or any small organic molecule inhibitor of such targets.

[0192] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-CTLA4 antibody.

[0193] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-CSl antibody.

[0194] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR2DLl / 2 / 3 antibody.

[0195] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-CD137 (e.g., urelumab) antibody.

[0196] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-GITR (e.g., TRX518) antibody.

[0197] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-PD-L2 antibody.

[0198] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILTl antibody.

[0199] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILT2 antibody.

[0200] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILT3 antibody.

[0201] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILT4 antibody.

[0202] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILT5 antibody.

[0203] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILT6 antibody.

[0204] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILT7 antibody.

[0205] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ILT8 antibody.

[0206] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-CD40 antibody.

[0207] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-OX40 antibody.

[0208] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR2DLl antibody.

[0209] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR2DL2 / 3 antibody.

[0210] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR2DL4 antibody.

[0211] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR2DL5A antibody.

[0212] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR2DL5B antibody.

[0213] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR3DLl antibody.

[0214] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR3DL2 antibody.

[0215] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-KIR3DL3 antibody.

[0216] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-NKG2A antibody.

[0217] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-NKG2C antibody.

[0218] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-ICOS antibody.

[0219] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-SIRPa antibody.

[0220] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-4-lBB antibody.

[0221] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-IL-10 antibody.

[0222] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with an anti-TSLP antibody.

[0223] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with IL- 10 or PEGylated IL- 10.

[0224] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with one or more of an inhibitor (e.g., a small organic molecule or an antibody or antigen-binding fragment thereof) such as: an MTOR (mammalian target of rapamycin) inhibitor, a cytotoxic agent, a platinum agent, an EGFR inhibitor, a VEGF inhibitor, a microtubule stabilizer, a taxane, a CD20 inhibitor, a CD52 inhibitor, a CD30 inhibitor, a RANK (Receptor activator of nuclear factor kappa-B) inhibitor, a STING agonist, a CXCR2 antagonist, a RANKL (Receptor activator of nuclear factor kappa-B ligand) inhibitor, an ERK inhibitor, a MAP Kinase inhibitor, an AKT inhibitor, a MEK inhibitor, a PARP inhibitor, a PI3K inhibitor, a HER1 inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, a Bcl2 inhibitor, a CD22 inhibitor, a CD79b inhibitor, an ErbB2 inhibitor, or a famesyl protein transferase inhibitor.

[0225] In some examples, the anti-CD27 antibody or antigen binding fragment thereof is coadministered with one or more chemotherapeutic agents. Any suitable chemotherapeutic agent, including any chemotherapeutic agent or combination thereof disclosed herein, may be used. In some examples, the one or more chemotherapeutic agents comprise nab-paclitaxel. In some examples, the nab-paclitaxel is administered to the patient via intravenous infusion. In some examples, the nab-paclitaxel is administered to the patient at a dose of about 100 mg / m2In some examples, the nab-paclitaxel is administered to the patient on a 28-day cycle, and the nab- paclitaxel is administered to the patient on Day 1, 8, and 15 of the 28-day cycle. In some examples, the nab-paclitaxel is administered to the patient on a 28-day cycle, and the nab-pachtaxel is administered to the patient at a dose of about 100 mg / m2on Day 1, 8, and 15 of the 28-day cycle.

[0226] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with any one or more of: 13-cis-retinoic acid, 3-[5- (methylsulfonylpiperadinemethyl)-indolyl]-quinolone, 4-hydroxytamoxifen, 5- deooxyuridine, 5'-deoxy-5-fluorouridine, 5 -fluorouracil, 6-mecaptopurine, 7-hydroxystaurosporine, A-443654, abirateroneacetate, ABT-578, acolbifene, ADS- 100380, ALT-110, altretamine, amifostine, aminoglutethimide, amrubicin, Amsacrine, anagrelide, anastrozole, angiostatin, AP-23573, ARQ- 197, arzoxifene, AS-252424, AS- 605240. asparaginase, AT-9263, atrasentan, axitinib, AZDI 152, Bacillus Calmette-Guerin (BCG) vaccine, batabulin, BC-210, besodutox, bevacizumab, bicalutamide, Biol 11, BIO140, bleomycin, BMS-214662, BMS-247550, BMS- 275291, BMS-310705, bortezimib, buserelin, busulfan, calcitriol, camptothecin, canertinib, capecitabine, carboplatin, carmustine, CC8490, cediranib, CG-1521, CG-781, chlamydocin, chlorambucil, chlorotoxin, cilengitide. cimitidine. cisplatin, cladribine, clodronate. COL-3, CP- 724714, cyclophosphamide, cyproterone, cyproteroneacetate, cytarabine, cytosinearabinoside, dacarbazine, dacinostat, dactinomycin, dalotuzumab, danusertib, dasatanib, daunorubicin, decatanib, deguelin, denileukin, deoxycoformycin, depsipeptide, diarylpropionitrile, diethylstilbestrol, diftitox, docetaxel, dovitinib, doxorubicin, droloxifene. edotecarin. yttrium-90 labeled-edotreotide, edotreotide, EKB-569, EMD121974, endostatin, enzalutamide, enzastaurin, epirubicin, epithilone B, ERA-923, cetuximab (Erbitux®), erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, ficlatuzumab, finasteride, flavopiridol, floxuridine, fludarabine, fludrocortisone, fluoxymesterone, flutamide, FOLFOX regimen, fulvestrant, galeterone. gefitinib. gemcitabine, gimatecan. goserelin, goserehn acetate, gossypol.GSK461364, GSK690693, HMR-3339, hydroxyprogesteronecaproate, hydroxyurea, IC87114, idarubicin, idoxyfene, ifosfamide, IM862, imatinib, IMC-1C11, INCB24360, INO1001, interferon, interleukin- 12, ipilimumab, irinotecan, JNJ-16241199, ketoconazole, KRX-0402, lapatinib. lasofoxifene. letrozole, leucovorin, leuprolide, leuprolide acetate, levamisole, liposome entrapped paclitaxel, lomustine, lonafamib, lucanthone, LY292223, LY292696, LY293646, LY293684, LY294002, LY317615, marimastat, mechlorethamine, medroxyprogesteroneacetate, megestrolacetate, melphalan, mercaptopurine, mesna, methotrexate, mithramycin, mitomycin, mitotane. mitoxantrone, tozasertib, MLN8054, neovastat, Neratinib, neuradiab, nilotinib, nilutimide, nolatrexed, NVP-BEZ235, oblimersen, octreotide, ofatumumab, olaparib, oregovomab, orteronel, oxaliplatin, paclitaxel, palbociclib, pamidronate, panitumumab, pazopanib, PD0325901, PD184352, PEG-interferon, pemetrexed, pentostatin, perifosine,phenylalanine mustard, Pl-103, pictilisib, P1K.-75. pipendoxifene. PKJ-166, plicamycin, porfimer, prednisone, procarbazine, a progestin, PX-866, R-763, raloxifene, raltitrexed, razoxin, ridaforolimus, rituximab, romidepsin, RTA744, rubitecan, scriptaid, Sdxl02, seliciclib, selumetinib, semaxanib, SF1126, sirolimus, SN36093, sorafenib, spironolactone, squalamine, SRI 3668, streptozocin, SU6668, suberoylanalide hydroxamic acid, sunitinib, synthetic estrogen, talampanel, talimogene laherparepvec, tamoxifen, temozolomide, temsirolimus, teniposide, tesmilifene, testosterone, tetrandrine, TGX-221, thalidomide, thioguanine, thiotepa, ticilimumab, tipifamib, tivozanib, TKI-258, TLK286, topotecan, toremifene citrate, trabectedin, trastuzumab, tretinoin, trichostatin A, triciribinephosphate monohydrate, triptorelin pamoate, TSE-424, uracil mustard, valproic acid, valrubicin, vandetanib, vatalanib, VEGF trap, vinblastine, vincristine, vindesine, vinorelbine, vitaxin, vitespan, vorinostat, VX-745, wortmannin, Xr311, zanolimumab, ZK186619, ZK-304709, ZM336372, or ZSTK474.

[0227] Additional non-limiting examples of suitable anti-cancer agents that may be used in combination with an anti-CD27 antibody or antigen-binding fragment thereof include cytostatic agents, cytotoxic agents, targeted therapeutic agents (e.g., small molecules, biologies, siRNA and microRNA) against cancer and neoplastic diseases, anti -metabolites (such as methotrexate, 5- fluorouracil, gemcitabine, fludarabine, capecitabine); alkylating agents, such as temozolomide, cyclophosphamide; DNA interactive and DNA damaging agents, such as cisplatin, oxaliplatin, and doxorubicin; Ionizing irradiation, such as radiation therapy: topoisomerase II inhibitors, such as etoposide and doxorubicin; topoisomerase I inhibitors, such as irinotecan and topotecan; tubulin interacting agents, such as paclitaxel, docetaxel, and nab-paclitaxel (Abraxane®), epothilones; kinesin spindle protein inhibitors; spindle checkpoint inhibitors; poly(ADP-ribose) polymerase (PARP) inhibitors, such as olaparib, niraparib and veliparib; matrix metalloprotease (MMP) inhibitors; protease inhibitors, such as cathepsin D and cathepsin K inhibitors; proteosome or ubiquitination inhibitors, such as bortezomib; adenoviral-p53; Bcl-2 inhibitors, such as ABT-263; heat shock protein (HSP) modulators, such as geldanamycin and 17-AAG; histone deacetylase (HD AC) inhibitors, such as vorinostat (SAHA); sex hormone modulating agents, such as anti-estrogens (e.g., tamoxifen and fulvestrant), selective estrogen receptor modulators (SERM) (e.g., raloxifene), anti-androgens (e.g., bicalutamide, flutamide), and luteinizing hormone-releasing hormone (LHRH) agonists (e.g., leuprolide); 5a-reductase inhibitors, such as finasteride; cytochrome P450 C17 lyase (CYP450cl7. also called 17aC); aromatase inhibitors, such as letrozole, anastrozole, and exemestane; EGFR kinase inhibitors, such as gefitinib, erlotinib, and lapatinib; multi-targeted kinases (serine / threonine and / or tyrosine kinase) inhibitors; ABL kinase inhibitors, such as imatinib, nilotinib, and dasatinib; VEGFR-1,VEGFR-2. PDGFR, K.DR. FLT. c-Kit, Tie2, Raf. MEK or ERK inhibitors, such as sunitinib. sorafenib, vandetanib, pazopanib, PLX-4032, axitinib, PTK787, or GSK-1120212; Polo-like kinase inhibitors; Aurora kinase inhibitors; JAK inhibitors; c-MET kinase inhibitors; PI3K and mTOR inhibitors, such as GDC-0941, BEZ-235, BKM-120 and AZD-8055; rapamycin and its analogs, such as temsirolimus. everolimus, and deforolimus; a STING (Stimulator of Interferon Genes) agonist; a CXCR (CXC Chemokine Receptor) inhibitor; a CXCR2 antagonist, other anticancer (also known as anti-neoplastic) agents including but are not limited to ara-C, adriamycin, cytoxan, carboplatin. uracil mustard, clormethine, ifosfsmide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatine, vinblastine, vincristine, vindesine, vinorelbine, navelbine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, teniposide, cytarabine, pemetrexed, idarubicin, mithramycin, deoxy coformycin, mitomycin-C, L-asparaginase, teniposide. ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, flutamide medroxyprogesteroneacetate, toremifene, goserelin, hydroxyurea, amsacrine. procarbazine, mitotane, mitoxantrone. levamisole, drolloxafme, hexamethylmelamine, tositumomab (Bexxar®), ibritumomab tiuxetan, arsenic trioxide, porfimer sodium, thiotepa, altretamine, liposomal doxorubicin, denileukin diftitox (Ontak®), cytarabine liposome injection (Depocyt®), darbepoetin alfa (Aranesp®), filgrastim (Neupogen®), pegfilgrastim (Neulasta®), or palifermin (Kepivance); famesyl protein transferase inhibitors, such as SARAS AR™, tipifamib; interferons, such as Intron A. Peg-Intron; anti-ErbBl antibodies, such as cetuximab, panitumumab; anti-ErbB2 antibodies, such as trastuzumab; anti- CD52 antibodies, such as alemtuzumab, anti-CD20 antibodies, such as rituximab; anti-CD33 antibodies, such as gemtuzumab ozogamicin; anti-VEGF antibodies, such as bevacizumab (Avastin®), TRAIL ligands, such as lexatumumab, mapatumumab. and AMG-655; anti-CTLA-4 antibodies, such as ipilimumab; antibodies against CTA1, CEA, CD5, CD19, CD22, CD30, CD44, CD44V6, CD55, CD56, EpCAM, FAP, MHCII, HGF, IL-6, MUC1, PSMA, TAL6, TAG- 72, TRAILR, VEGFR, IGF-2, FGF; or anti-IGF-lR antibodies, such as dalotuzumab and robatumumab (SCH 717454).

[0228] Examples of cytotoxic / cytostatic agents that may be used in association with an anti- CD27 antibody or an antigen-binding fragment thereof include, but are not limited to, platinum coordinator compounds, sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin,altretamine, prednimustine, dibromodulcitol, ranimustine. fotemustine. nedaplatin. oxaliplatin, temozolomide, heptaplatin, estramustine, improsulfan tosilate, trofosfamide, nimustine, dibrospidium chloride, pumitepa, lobaplatin, satraplatin, profiromycin, cisplatin, irofulven, dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum, benzylguanine, glufbsfamide, GPX100. (trans, trans, trans)-bis-mu-(hexane-1.6-diamine)-mu-[diamine- platinum(II)]bis[diamine(chloro)platinum (II)]tetrachloride, diarizidinylspermine, arsenic trioxide, 1-(1 l-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin, pinafide, valrubicin, amrubicin, antineoplaston. 3'-deamino-3'-morpholino-l 3-deoxo-I O-hydroxycarminomycin. annamycin, galarubicin, elinafide, MEN10755, 4-demethoxy-3-deamino-3-aziridinyl-4-methylsulphonyl- daunorubicin (see WO 00 / 50032).

[0229] An example of a hypoxia activatable compound that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof include, but are not limited to, is tirapazamine.

[0230] Examples of proteosome inhibitors that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof include, but are not limited to, include but are not limited to lactacystin and bortezomib (Velcade®).

[0231] Examples of microtubule inhibitors / microtubule-stabilizing agents that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof include taxanes in general. Specific compounds include paclitaxel (Taxol®), nab-paclitaxel (Abraxane®), vindesine sulfate, 3’,4'-didehydro-4’-deoxy-8’-norvincaleukoblastine, docetaxel (Taxotere®), rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin, RPR109881, BMS184476. vinflunine. cryptophycin, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl) benzene sulfonamide, anhydrovinblastine, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L- proline-t-butylamide, TDX258, the epothilones (see for example U.S. Pat. Nos. 6,284,781 and 6,288,237) and BMS188797.

[0232] Some examples of topoisomerase inhibitors that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof include, but are not limited to, are topotecan, hycaptamine, irinotecan, rubitecan, 6-ethoxypropionyl-3’,4’-O-exo-benzy lidene- chartreusin, 9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H) propanamine, 1- amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-114,12H-benzo[de]pyrano[3’,4’:b,7]- indolizinof 1,2b] quinoline- 10,13(9H, 15H)di one, lurtotecan, 7-[2-(N-isopropylamino)ethyl]- (20S)camptothecin, BNP1350, BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane, 2’-dimethylamino-2’-deoxy-etoposide, GL331, N-[2-(dimethylamino)ethyl]-9-hydroxy-5.6-dimethyl-6H-pyrido[4.3-b]carbazole-l -carboxamide, asulacnne, (5a, 5aB, 8aa.9b)- 9- [2- [N- [2-(dimethy lamin o)ethy 1] -N-methylamino] ethyl] -5- [4- hy droOxy-3 ,5 -dimethoxypheny 1] - 5,5a,6,8,8a,9-hexohydrofuro(3’,4’:6,7)naphtho(2,3-d)-l,3- dioxol-6-one, 2,3-(methylenedioxy)- 5-methyl-7-hydroxy-8-methoxybenzo[c]- phenanthridinium, 6,9-bis[(2- aminoethyl)amino]benzo|g]isoguinoline-5, 10-dione, 5-(3-aminopropylamino)-7.10-dihydroxy-2- (2 -hydroxy ethylaminomethyl)-6H-pyrazolo[4,5, 1 - de]acridin-6-one, N-[l - [2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4- ylmethyl] formamide, N-(2- (dimethy lamino)ethyl)acridine-4-carboxamide, 6-[[2-(dimethylamino)ethy 1] amino] -3-hy droxy- 7H-indeno[2,l-c] quinolin-7-one, and dimesna.

[0233] Examples of inhibitors of mitotic kinesins, and in particular the human mitotic kinesin KSP, that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof, are described in W003 / 039460, W003 / 050064, W003 / 050122, WO03 / 049527, WO03 / 049679, WO03 / 049678, WO04 / 039774, WO03 / 079973, W003 / 099211, W003 / 105855, W003 / 106417, W004 / 037171, W004 / 058148. W004 / 058700, WO04 / 126699, W005 / 018638. W005 / 019206, W005 / 019205, W005 / 018547, W005 / 017190, US2005 / 0176776.

[0234] Examples of histone deacetylase inhibitors that may be used in association with an anti- CD27 antibody or an antigen-binding fragment thereof include, but are not limited to, SAHA, TSA. oxamflatin, PXD101, MG98 and scriptaid. Further reference to other histone deacetylase inhibitors may be found in Miller, T.A. et al. J. Med. Chem. 46(24):5097-5116 (2003).

[0235] Examples of inhibitors of kinases involved in mitotic progression that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof include, but are not limited to, inhibitors of aurora kinase, inhibitors of Polo-like kinases (PLK; in particular inhibitors of PLK- 1), inhibitors of Bub- 1 and inhibitors of Bub-Rl. An example of an aurora kinase inhibitor is VX-680.

[0236] Antiproliferative agents that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof includes antisense RNA and DNA oligonucleotides such as G3139. ODN698, RVASKRAS. GEM231, and INX3001, and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin, doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine, cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed, paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed, nelzarabine, 2’-deoxy-2’-methylidenecytidine, 2‘- fluoromethylene-2’ -deoxy cytidine, N-[5-(2,3-dihydro-benzofuryl)sulfonyl]-N’-(3,4- dichlorophenyl)urea, N6-[4-deoxy-4-[N2-[2(E),4(E)- tetradecadienoyl]glycylamino]-L-glycero- B-L-manno-heptopyranosyl]adenine, aplidine, ecteinascidin, troxacitabine, 4-[2-amino-4-oxo- 4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][l,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamicacid, aminopterin, 5-flurouraciL alanosine, l l-acetyl-8-(carbamoyloxymethyl)-4-formyl-6- meth oxy-14-oxa-l,l 1- diazatetracyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-yl acetic acid ester, swainsonine, lometrexol, dexrazoxane, methioninase, 2’-cyano-2’-deoxy-N4-palmitoyl-l-B-D- arabino furanosyl cytosine, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone and trastuzumab.

[0237] Examples of angiogenesis inhibitors that may be used in association with an anti-CD27 antibody or an antigen-binding fragment thereof include, but are not limited to, tyrosine kinase inhibitors, such as inhibitors of the tyrosine kinase receptors Fit- 1 (VEGFR1) and Flk-1 / KDR (VEGFR2), inhibitors of epidermal-derived, fibroblast-derived, or platelet derived growth factors, MMP (matrix metalloprotease) inhibitors, integrin blockers, interferon-a, interleukin- 12, pentosan polysulfate, cyclooxygenase inhibitors, including nonsteroidal anti-inflammatories (NSAIDs) like aspirin and ibuprofen as well as selective cyclooxy -genase-2 inhibitors like celecoxib and rofecoxib (PNAS. Vol. 89, p. 7384 (1992); JNCI, Vol. 69, p. 475 (1982); Arch. Opthalmol., Vol. 108, p.573 (1990); Anat. Rec.. Vol. 238, p. 68 (1994); FEBS Letters. Vol. 372. p. 83 (1995); Clin, Orthop. Vol. 313, p. 76(1995); J. Mol. Endocrinol., Vol. 16, p.107 (1996); Jpn. J. Pharmacol., Vol. 75, p. 105 (1997); Cancer Res., Vol. 57, p. 1625 (1997); Cell, Vol. 93, p. 705 (1998); Inti. J. Mol. Med., Vol. 2, p. 715 (1998); J. Biol. Chem, Vol. 274, p. 9116 (1999)), steroidal anti-inflammatories (such as corticosteroids, mineralocorticoids, dexamethasone, prednisone, prednisolone, methylpred, betamethasone), carboxyamidotriazole, combretastatin A- 4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin, troponin-1, angiotensin II antagonists (see Fernandez et al., J. Lab. Clin. Med. 105:141-145 (1985)), and antibodies to VEGF (see, Nature Biotechnology, Vol. 17, pp.963-968 (October 1999); Kim et al., Nature. 362. 841-844 (1993); WO 00 / 44777; and WO 00 / 61186).

[0238] Other examples of angiogenesis inhibitors include, but are not limited to, endostatin, ukrain, ranpimase, IM862, 5-methoxy-4-[2-methyl-3-(3-methyl-2-butenyl)oxiranyl]-l- oxaspiro[2.5]oct-6-yl(chloroacetyl)carbamate. acetyl dinanaline, 5-amino-l-[[3,5-dichloro-4-(4- chlorobenzoyl)phenyl]methyl]- 1 H- 1 ,2,3-triazole-4-carboxamide,CMl 01. squalamine, combretastatin, RPI4610, NX31838, sulfated mannopentaose phosphate, 7,7-(carbonyl- bis[imino-N-methyl-4,2-pyrrolocarbonylimino[N-methyl-4,2-pyrrole]-carbonylimino]-bis- (1,3- naphthalene disulfonate), and 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone (SU5416).

[0239] Some specific examples of tyrosine kinase inhibitors include N-(trifluoromethylphenyl)- 5-methylisoxazol-4-carboxamide, 3-[(2,4-dimethylpyrrol-5- yl)methylidenyl)indolin-2-one, 17- (allylamino)-17-demethoxygeldanamycin, 4-(3-chloro-4- fluorophenylamino)-7-methoxy-6-[3- (4-morpholinyl)propoxyl]quinazoline, N-(3- ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, B1BX1382, 2.3,9,10,1 1.12- hexahydro-10-(hydroxymethyl)-10-hydroxy-9- methyl-9,12-epoxy-lH-diindolo[l,2,3-fg:3’,2’,r-kl]pyrrolo[3,4-i][l,6]benzodiazocin-l-one, SH268, genistein, STI571, CEP2563, 4-(3-chlorophenylamino)-5,6-dimethyl-7H-pyrrolo[2,3- d] pyrimidinemethane sulfonate, 4-(3- bromo-4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, 4-(4’-hydroxyphenyl)amino-6,7- dimethoxyquinazoline, SU6668, STI571A, N-4-chlorophenyl- 4-(4-pyridylmethyl)-l- phthal azinamine, and EMD121974.

[0240] In some examples, anti-CD27 antibodies or antigen binding fragments may be used in association with PPAR-y (i.e., PPAR-gamma) agonists or PPAR-5 (i.e., PPAR-delta) agonists. PPAR-y and PPAR-5 are the nuclear peroxisome proliferator-activated receptors y and 5. The expression of PPAR-y on endothelial cells and its involvement in angiogenesis has been reported in the literature (see Agarwal J. Cardiovasc. Pharmacol. 1998; 31:909-913; J. Biol. Chem. 1999;274:9116-9121; Murata et ak, Invest. Ophthalmol Vis. Sci. 2000; 41:2309-2317). PPAR-y agonists have been shown to inhibit the angiogenic response to VEGF in vitro; both troglitazone and rosiglitazone maleate inhibit the development of retinal neovascularization in mice. (Arch. Ophthamol. 2001; 119:709-717). Examples of PPAR-y agonists and PPAR-y / a agonists include, but are not limited to, Lynparza®, Rucaparib®, Talazoparib®, niraparib, Veliparib®, thiazolidinediones (such as DRF2725, CS-011, troglitazone, rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate, GW2570, SB219994. AR-H039242, JTT-501, MCC-555, GW2331, GW409544, NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, DRF552926, 2-[(5,7-dipropyl-3-trifluoromethyl-l,2-benzisoxazol-6-yl)oxy]-2-methylpropionic acid, and 2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy) phenoxy)propoxy)-2-ethylchromane-2- carboxylic acid.

[0241] Anti-CD27 antibodies or antigen-binding fragments thereof may also be used for treating or preventing breast cancer in association with aromatase inhibitors. Examples of aromatase inhibitors include but are not limited to: anastrozole, letrozole and exemestane.

[0242] Anti-CD27 antibodies or antigen-binding fragments thereof may also be used in association with one or more of the following chemotherapeutic agents: abarelix (Plenaxis depot®); aldesleukin (Prokine®); Aldesleukin (Proleukin®); alitretinoin (Panretin®); allopurinol (Zyloprim®); altretamine (Hexalen®); amifostine (Ethyol®); anastrozole (Arimidex®); arsenic trioxide (Trisenox®); asparaginase (Elspar®); azacitidine (Vidaza®); bendamustine hydrochloride (Treanda®); bexarotene capsules (Targretin®); bexarotene gel (Targretin®); bleomycin (Blenoxane®); bortezomib (Velcade®); brefeldin A; busulfan intravenous (Busulfex®); busulfan oral (Myleran®); calusterone (Methosarb®); capecitabine (Xeloda®); carboplatin (Paraplatin®); carmustine (BCNU®, BiCNU®); carmustine (Gliadel®); carmustinewi th Polifeprosan 20 Implant (Ghadel Wafer®); celecoxib (Celebrex®); chlorambucil (Leukeran®); cisplatin (Platinol®); cladribine (Leustatin®, 2-CdA®); clofarabine (Clolar®); cyclophosphamide (Cytoxan®, Neosar®); cyclophosphamide (Cytoxan Injection®); cyclophosphamide (Cytoxan Tablet®); cytarabine (Cytosar-U®); cytarabine liposomal (DepoCyt®); dacarbazine (DTIC-Dome®); dactinomycin, actinomycin D (Cosmegen®); dalteparin sodium injection (Fragmin®); darbepoetin alfa (Aranesp®); dasatinib (Spry cel®); daunorubicin liposomal (DanuoXome®); daunomycin (Daunorubicin®); daunorubicin, daunomycin (Cerubidine®); degarelix (Firmagon®); Denileukin diftitox (Ontak®); dexrazoxane (Zinecard®); dexrazoxane hydrochloride (Totect®); didemnin B; 17-DMAG; docetaxel (Taxotere®); doxorubicin (Adriamycin PFS®); doxorubicin (Adriamycin®, Rubex®); doxorubicin (Adriamycin PFS Injection®); doxorubicin liposomal (Doxil®); dromostanolone propionate (Dromostanolone ®); dromostanolone propionate (Masterone Injection®); eculizumab injection (Soliris®); Elliott's B Solution (Elliott's B Solution®); eltrombopag (Promacta®); epirubicin (Ellence®); Epoetin alfa (epogen®); erlotinib (Tarceva®); estramustine (Emcyt®); ethinyl estradiol; etoposide phosphate (Etopophos®); etoposide, VP- 16 (Vepesid®); everolimus tablets (Afinitor®); exemestane (Aromasin®); ferumoxytol (Feraheme Injection®); floxuridine (intraarterial) (FUDR®): fludarabine (Fludara®); fluorouracil, 5-FU (Adrucil®); fulvestrant (Faslodex®); gefitinib (Iressa®); geldanamycin; gemcitabine (Gemzar®); gemtuzumab ozogamicin (Mylotarg®); goserelin acetate (Zoladex Implant®); goserelin acetate (Zoladex®); histrelin acetate (Histrelin implant®); hydroxyurea (Hydreaidarubicin (Idamycin®); ifosfamide (IFEX®); imatinib mesylate (Gleevec®); interferon alfa 2a (Roferon A®); Interferon alfa-2b (Intron A®); iobenguane I123injection (AdreView®); irinotecan (Camptosar®); ixabepilone (Ixempra®); lapatinib tablets (Tykerb®); lenalidomide (Revhmid®); letrozole (Femara®); leucovorin (Wellcovorin®, Leucovorin®); Leuprolide Acetate (Eligard®); levamisole (Ergamisol®); lomustine, CCNU (CeeBU®); meclorethamine, nitrogen mustard (Mustargen®); megestrol acetate (Megace®); melphalan, L-PAM (Alkeran®); mercaptopurine, 6-MP (Purinethol®); mesna (Mesnex®); mesna (Mesnex tabs®); methotrexate (Methotrexate®); methoxsalen (Uvadex®); 8-methoxypsoralen; mitomycin C (Mutamycin®); mitotane (Lysodren®); mitoxantrone (Novantrone®); mitramycin; nandrolone phenpropionate (Durabolin- 50®); nelarabine (Arranon®); nilotinib (Tasigna®); Nofetumomab (Verluma®); ofatumumab (Arzerra®); Oprelvekin (Neumega®); oxaliplatin (Eloxatin®); paclitaxel (Paxene®); paclitaxel (Taxol®); paclitaxel protein-bound particles (Abraxane®); palifermin (Kepivance®); pamidronate (Aredia®); panitumumab (Vectibix®); pazopanib tablets (Votrienttm®); pegademase (Adagen (Pegademase Bovine)®); pegaspargase (Oncaspar®); pemetrexeddisodium (Alimta®); pentostatin (Nipent®); pipobroman (Vercyte®); plerixafor (Mozobil®); plicamycin, mithramycin (Mithracin®); porfimer sodium (Photofrin®); pralatrexate injection (Folotyn®); procarbazine (Matulane®); quinacrine (Atabrine®); rapamycin; rasburicase (Elitek®); raloxifene hydrochloride (Evista®); romidepsin (Istodax®); romiplostim (Nplate®); sargramostim (Leukine®); sargramostim (Prokine®); sorafenib (Nexavar®); streptozocin (Zanosar®); sunitinib maleate (Sutent®); talc (Sclerosol®); tamoxifen (Nolvadex®); temozolomide (Temodar®); temsirolimus (Torisel®); teniposide, VM-26 (Vumon®); testolactone (Teslac®); thioguanine, 6-TG (Thioguanine®); thiopurine; thiotepa (Thioplex®); topotecan (Hycamtin®); toremifene (Fareston®); Tositumomab (Bexxar®); Tositumomab / I-131 tositumomab (Bexxar®); trans-retinoic acid; tretinoin, ATRA (Vesanoid®); triethylenemelamine; uracil mustard (Uracil Mustard Capsules®); valrubicin (Valstar®); vinblastine (Velban®); vincristine (Oncovin®); vinorelbine (Navel bine®); vorinostat (Zolinza®); wortmannin; and zoledronate (Zometa®).

[0243] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with anti-cancer radiation therapy. For example, in an embodiment, the radiation therapy is external beam therapy (EBT): a method for delivering a beam of high-energy X-rays to the location of the tumor. The beam is generated outside the patient (e.g., by a linear accelerator) and is targeted at the tumor site. These X-rays can destroy the cancer cells and careful treatment planning allows the surrounding normal tissues to be spared. No radioactive sources are placed inside the patient's body. In an embodiment, the radiation therapy is proton beam therapy: a type of conformal therapy that bombards the diseased tissue with protons instead of X-rays. In an embodiment, the radiation therapy is conformal external beam radiation therapy: a procedure that uses advanced technology to tailor the radiation therapy to an individual's body structures. In an embodiment, the radiation therapy is brachytherapy: the temporary placement of radioactive materials within the body, usually employed to give an extra dose or boost of radiation to an area.

[0244] In an embodiment, an anti-CD27 antibody or antigen-binding fragment thereof of the disclosure is used in association with a surgical procedure such as surgical tumorectomy.

[0245] In some examples, the patient is infused with autologous T cells expanded ex vivo with anti-CD27 specific antibodies or antigen-binding fragments thereof. In another embodiment, the patient is administered autologous T cells in combination with the anti-CD27 specific antibodies or antigen-binding fragments thereof. In yet another embodiment, the patient is vaccinated with a cancer vaccine, and infused with autologous T cells expanded ex vivo with anti-CD27 specific antibodies or antigen-binding fragments thereof. The autologous T-cells can be autologousinfiltrating lymphocytes. T-cells transduced with high affinity T-cell receptors against tumor antigens or T cells transduced with chimeric antigen receptors composed of hybrid immunoglobulin lights chains with endo-domains of T-cell signaling molecules. See Kalos M. and June C. H., Immunity, 39, 2013. p49-60; Wu R. et al., Cancer J. 2012; 18(2): 160-175; and June. C. H„ J. Clin. Invest. 117: 1466-1476 (2007).Anti-CD27 Antibodies and Antigen-binding Fragments

[0246] The present disclosure provides anti-CD27 antibodies and antigen-binding fragments thereof having specified structural and / or functional features, methods of use of the antibodies or antigen-binding fragments thereof in the treatment or prevention of disease (e.g., cancer (e.g., a cancer having a PD-L1 CPS <10)), and compositions comprising the antibodies or antigenbinding fragments thereof for use in the treatment or prevention of disease (e.g., cancer having a PD-L1 CPS <10).

[0247] Any suitable anti-CD27 antibody (e.g.. boserolimab) may be used in the methods, compositions, uses, and kits disclosed herein. Examples of anti-CD27 antibodies as used herein include, but are not limited to, boserolimab, varlilumab (also known as CDX-1127), BMS- 986215, BNT313 / GEN1053, and hCD27.15. Anti-CD27 antibodies are further described, e.g., in WO 2018 / 058022, WO 2021 / 087016, WO2012 / 004367, W02015 / 016718. WO 2008 / 051424. WO 2011 / 130434, WO 2019 / 195452, WO 2023 / 031473, and CN 1 16262788.

[0248] In some examples, the anti-CD27 antibody or antigen-binding fragment thereof that is discussed herein (e.g., hCD27.131A or humanized versions thereof disclosed in PCT publication number WO2018 / 058022) or a variant thereof (e.g.. a sequence variant or functional variant); any antibody or antigen-binding fragment thereof comprising any one or more of the CDRs set forth below in Table 3.

[0249] In some examples, the anti-CD27 antibody or the antigen-binding fragment thereof may include amino acid sequences of boserolimab. In some examples, the anti-CD27 antibody or the antigen-binding fragment thereof may include amino acid sequences set forth in Table 3.Table 3. Exemplary amino acid sequences of boserolimab

[0250] As stated above, antibodies and fragments thereof that bind to the same epitope as any of the anti-CD27 antibodies or antigen-binding fragments thereof described herein may be used as part of the present disclosure. In one embodiment, the disclosure provides an antibody or antigen binding fragment thereof that binds to the same epitope of human CD27 as an antibody or antigen binding fragment thereof described herein, for example, an antibody or antigen binding fragment thereof comprising the variable heavy chain comprising the amino acid sequence of SEQ ID NO:7 and the variable light chain comprising the amino acid sequence of SEQ ID NO:9. In another embodiment, the disclosure provides an antibody or antigen binding fragment thereof that binds to the same epitope of human CD27 as an antibody comprising the variable heavy chain comprising the amino acid sequence of SEQ ID NO: 7 and the variable light chain comprising the amino acid sequence of SEQ ID NO:9.

[0251] There are several methods available for mapping antibody epitopes on target antigens, including: H / D-Ex Mass spec, X-ray crystallography, pepscan analysis, alanine scanning, hydroxyl radical footprinting and site directed mutagenesis. For example, HDX (Hydrogen Deuterium Exchange) coupled with proteolysis and mass spectrometry can be used to determine the epitope of an antibody on a specific antigen Y. HDX-MS relies on the accurate measurement and comparison of the degree of deuterium incorporation by an antigen when incubated in D2O on its own and in presence of its antibody at various time intervals. Deuterium is exchanged with hydrogen on the amide backbone of the proteins in exposed areas whereas regions of the antigen bound to the antibody will be protected and will show less or no exchange after analysis by LC- MS / MS of proteolytic fragments. In one embodiment, the epitope is determined by solving the X-ray crystal structure of a complex between CD27 or fragment thereof and an anti-CD27 antibody or fragment thereof and identifying one or more CD27 residues within 4 A of the anti- CD27 antibody residues. In another embodiment, the epitope includes for example, CD27 residues that have van der Waals, polar interaction, salt bridge or hydrogen bond contact with the anti-CD27 antibody residues. In another embodiment, the epitope is determined by mutagenesis (for example Alanine scanning) of CD27 residues and analyzing the loss of binding to the anti- CD27 antibody as a result of the mutagenesis.

[0252] Also provided are isolated polypeptides comprising the VL domains of the anti-CD27 antibodies of the disclosure (e.g., SEQ ID NO: 9), and isolated polypeptides comprising the VH domains of the anti-CD27 antibodies of the disclosure (e.g., SEQ ID NO:7) having up to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid substitutions. In some examples, the substitutions are in the framework regions. In other examples, the substitutions are in the CDR regions. In other examples, the substations are in the CDR regions and the framework regions.

[0253] In another embodiment, an antibody or antigen-binding fragment thereof that binds CD27 and has VL domains and VH domains with at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, 80% or 75% sequence identity to one or more of the VL domains or VH domains described herein and exhibits specific binding to CD27 may also be utilized in the methods and uses described herein. In another embodiment, the binding antibody or antigen-binding fragment thereof of the present disclosure comprises VL and VH domains (with and without signal sequence) having up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more amino acid substitutions, and exhibits specific binding to CD27.

[0254] Also included in the present disclosure are polypeptides, e.g., immunoglobulin polypeptides, comprising amino acid sequences that are at least about 75% identical, 80% identical, more preferably at least about 90% identical and most preferably at least about 95%identical (e.g., 95%, 96%, 97%, 98%, 99%, 100%) to the amino acid sequences of the antibodies provided herein when the comparison is performed by a BLAST algorithm wherein the parameters of the algorithm are selected to give the largest match between the respective sequences over the entire length of the respective reference sequences (e.g., expect threshold: 10; word size: 3; max matches in a query range: 0; BLOSUM 62 matrix; gap costs: existence 11, extension 1; conditional compositional score matrix adjustment).

[0255] In some embodiments, the methods, compositions, uses, and kits for treating cancer comprise administering to the subject an antibody or antigen binding fragment thereof that increases the activity of an immune cell. The increase of the activity of an immune cell can be detected using any method know n in the art. In one embodiment, the increase in activity of an immune cell can be detected by measuring the proliferation of the immune cell.

[0256] By way of example, and not limitation, methods, compositions, uses, and kits for treating cancer as disclosed herein may comprise administering the antibodies and antigenbinding fragments thereof disclosed herein (e.g., comprising an amino acid sequence set forth in Table 1) that may bind human CD27 or CD27A59T) with a bivalent KD value of 10 x 10‘9M or lower as determined by surface plasmon resonance (e.g., BIACORE®) or a similar technique (e.g., KINEXA® kinetic exclusion assay or OCTET® bio-layer interferometry) as measured with a human CD27-Fc fusion protein or human CD27A59T-Fc fusion protein. In one embodiment, methods and uses for treating cancer disclosed herein may comprise administering the antibodies and antigen-binding fragments thereof disclosed herein, wherein the antibodies or antigen binding fragments may bind human CD27 or CD27A59T with a bivalent KD value of about 5 x 10’9to about 10 x 10'9M as determined by surface plasmon resonance (e.g., BIACORE®) or a similar technique (e.g.. KINEXA® kinetic exclusion assay or OCTET® bio-layer interferometry) as measured using a CD27 protein or peptide, for example with a human CD27-Fc fusion protein or human CD27A59T-Fc fusion protein.

[0257] Bispecific and bifunctional antibodies and antigen-binding fragments thereof having a binding specificity for CD27 and another antigen such as. for example. PD-1 or PD-L1, and methods of use thereof are described herein. In an embodiment of the disclosure, the bispecific antibody may comprise an anti-CD27 antibody or antigen-binding fragment thereof comprising any of the sequences described in Table 3. and an anti-PD-1 antibody or antigen-binding fragment thereof comprising any of the sequences described in Table 4.

[0258] The present disclosure further includes methods, compositions, uses, and kits comprising administering an anti-CD27 antigen-binding fragment of an anti-CD27 antibody disclosed herein. Examples of antigen-binding fragments that may be used include, but are notlimited to, Fab. Fab’. F(ab’)2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules, e g., sc-Fv; and multispecific antibodies formed from antibody fragments.

[0259] In some embodiments, the anti-CD27 antibody or antigen-binding fragment thereof comprises a heavy chain constant region of the IgGl subtype.Anti-PD-1 Antibodies and Antigen-binding Fragments

[0260] Examples of mAbs that bind to human PD-1, useful in the treatment methods and uses of the disclosure, are described in US 7,521,051, US 8,008,449, and US 8,354,509. Specific antihuman PD-1 mAbs useful as a PD-1 antagonist in the treatment methods, compositions, uses, and kits of the present disclosure include: pembrohzumab (formerly known as MK-3475, SCH 900475 and lambrolizumab), a humanized IgG4 mAb with the structure described in WHO Drug Information, Vol. 27, No. 2, pages 161-162 (2013) and which comprises the heavy and light chain amino acid sequences shown in Table 4, nivolumab, cemiplimab. dostarlimab, retifanlimab. toripalimab, and the humanized antibodies h409Al 1, h409A16 and h409A17, which are described in WO 2008 / 156712.Table 4. Exemplary Antibody Sequences for Pembrolizumab

[0261] In some embodiments of the methods, compositions, uses, and kits of the present disclosure, the anti-PD-1 antibody, or antigen binding fragment thereof, comprises one or more (e g., 1, 2, 3, 4, 5, or all six) of the following: (a) light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences as set forth in SEQ ID NOs: 11, 12 and 13, respectively, and heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences as set forth in SEQ ID NOs: 16, 17 and 18. respectively.

[0262] In some embodiments of the methods, compositions, uses, and kits of the present disclosure, the anti-PD-1 antibody, or antigen binding fragment thereof, comprises: (a) light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences as set forth in SEQ ID NOs: 11, 12 and 13, respectively , and heavy chain CDR1, CDR2. and CDR3 comprising the amino acid sequences as set forth in SEQ ID NOs: 16, 17 and 18. respectively.

[0263] In some embodiments, the anti-PD-1 antibody or antigen binding fragment thereof is a human antibody. In other embodiments, the anti-PD-1 antibody or antigen binding fragment thereof is a humanized antibody. In other embodiments, the anti-PD-1 antibody or antigen binding fragment thereof is a chimeric antibody. In specific embodiments, the anti-PD-1 antibody or antigen binding fragment thereof is a monoclonal antibody.

[0264] In other embodiments of the treatment methods, compositions, kits and uses of the present invention, the anti-PD-1 antibody, or antigen binding fragment thereof, specifically binds to human PD-1 and comprises (a) a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 19. or a variant thereof, and (b) a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 14 or a variant thereof.

[0265] In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is a monoclonal antibody which specifically binds to human PD-1 and comprises (a) a heavy chain comprising or consisting of a sequence of amino acids as set forth in SEQ ID NO:20, or a variant thereof; and (b) a light chain comprising or consisting of a sequence of amino acids as set forth in SEQ ID NO: 15, or a variant thereof.

[0266] In some examples, a variant of a heavy chain variable region sequence or full-length heavy chain sequence is identical to the reference sequence except having one or more and up to 17 conservative amino acid substitutions in the framework region (i.e., outside of the CDRs), and in specific embodiments has less than ten, nine, eight, seven, six or five conservative amino acid substitutions in the framework region. A variant of a light chain variable region sequence or full- length light chain sequence is identical to the reference sequence except having one or more and up to five conservative amino acid substitutions in the framework region (i.e., outside of the CDRs), and in specific embodiments has less than four, three or two conservative amino acid substitution in the framework region.

[0267] In yet another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof is a monoclonal antibody which specifically binds to human PD-1 and comprises (a) a heavy chain comprising or consisting of a sequence of amino acids as set forth in SEQ ID NO:20 and (b) a light chain comprising or consisting of a sequence of amino acids as set forth in SEQ ID NO:15.

[0268] The present disclosure further includes methods, compositions, uses, and kits comprising administering an anti-PD-1 antigen-binding fragment of an anti-PD-1 antibody disclosed herein. Examples of antigen-binding fragments that may be used include, but are not limited to, Fab. Fab’. F(ab’)2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules, e g., sc-Fv; and multispecific antibodies formed from antibody fragments.Polynucleotides

[0269] The present disclosure further provides polynucleotides encoding any of the polypeptides or immunoglobulin chains of antibodies (e.g., anti-CD27 antibodies or anti-PD-1 antibodies) and antigen-binding fragments thereof of the disclosure. For example, the present disclosure includes polynucleotides encoding an anti-CD27 antibody or antigen-binding fragment thereof having the amino acids described in Table 3. In another example, the present disclosure includes polynucleotides encoding an anti-PD-1 antibody or antigen-binding fragment thereof having the amino acids described in Table 4.

[0270] In one embodiment, an isolated polynucleotide, for example DNA, encoding the polypeptide chains of the isolated antibodies or antigen-binding fragments thereof set forth herein is provided. In one embodiment, the isolated polynucleotide encodes an antibody or antigenbinding fragment thereof comprising at least one mature immunoglobulin light chain variable (VL) domain according to the disclosure and / or at least one mature immunoglobulin heavy chain variable (VH) domain according to the disclosure. In some embodiments the isolated polynucleotide encodes both a light chain and a heavy chain on a single polynucleotide molecule, and in other embodiments the light and heavy chains are encoded on separate polynucleotide molecules. In another embodiment the polynucleotide further encodes a signal sequence.

[0271] In one embodiment, the disclosure provides an isolated polynucleotide encoding a VH domain or an antigen-binding fragment thereof comprising CDR-H1 (SEQ ID NO:1), CDR-H2 (SEQ ID NO:2) and CDR-H3 (SEQ ID NO:3). In one embodiment, the disclosure provides an isolated polynucleotide encoding a VL domain or an antigen-binding fragment thereof comprising CDR-L1 (SEQ ID NO:4), CDR-L2 (SEQ ID NO:5) and CDR-L3 (SEQ ID NO:6). In one embodiment, the disclosure provides an isolated polynucleotide encoding the VH domain comprising the amino acid sequence of SEQ ID NO: 7. In one embodiment, the disclosure provides an isolated polynucleotide encoding the VL domain comprising the amino acid sequence of SEQ ID NOV. In one embodiment, the disclosure provides an isolated polynucleotide encoding the heavy chain comprising the amino acid sequence of SEQ ID NO: 8. In one embodiment, the disclosure provides an isolated polynucleotide encoding the light chain comprising the amino acid sequence of SEQ ID NO: 10.

[0272] The present disclosure also provides vectors, e.g., expression vectors, such as plasmids, comprising the isolated polynucleotides of the disclosure, wherein the polynucleotide is operably linked to control sequences that are recognized by a host cell when the host cell is transfected with the vector. Also provided are host cells comprising a vector of the present disclosure and methods for producing an antibody or antigen-binding fragment thereof or polypeptide disclosed herein (e.g., in Table 3 or Table 4) comprising culturing a host cell harboring an expression vector or a nucleic acid encoding the immunoglobulin chains of the antibody or antigen-binding fragment thereof in culture medium, and isolating the antigen or antigen-binding fragment thereof from the host cell or culture medium.Methods of Making Antibodies and Antigen-binding Fragments Thereof

[0273] Methods for making antibodies (e.g., anti-CD27 antibodies or anti-PD-1 antibodies) and antigen-binding fragments thereof are described herein. For example, the disclosure providesmethods comprising culturing a hybndoma cell that expresses the antibody or fragment thereof under conditions favorable to such expression and, optionally, isolating the antibody or fragment thereof from the hybridoma and / or the growth medium (e.g., cell culture medium).

[0274] The antibodies disclosed herein may also be produced recombinantly (e g., in an E. coli / Tl expression system, a mammalian cell expression system, or a lower eukaryote expression system). Nucleic acids encoding the antibody immunoglobulin molecules of the disclosure (e g., VH and / or VL) may be inserted into a pET-based plasmid and expressed in the E. coliEVl system. For example, methods for expressing an antibody or antigen-binding fragment thereof or immunoglobulin chain thereof in a host cell (e.g., bacterial host cell such as E.coli such as BL21 or BL21DE3) are described herein. In various embodiments, the methods comprise expressing T7 RNA polymerase in the cell which also includes a polynucleotide encoding an immunoglobulin chain that is operably linked to a T7 promoter. For example, a bacterial host cell, such as an E. coll cell, includes a polynucleotide encoding the T7 RNA polymerase gene operably linked to a lac promoter and expression of the polymerase and the chain is induced by incubation of the host cell with isopropyl-beta-D-thiogalactopyranoside (IPTG).

[0275] There are several methods by which to produce recombinant antibodies which are known in the art. One example of a method for recombinant production of antibodies is disclosed in U.S. Patent No. 4.816,567.

[0276] Transformation can be by any known method for introducing polynucleotides into a host cell. Methods for introduction of heterologous polynucleotides into mammalian cells are w ell known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, biolistic injection and direct microinjection of the DNA into nuclei. In addition, nucleic acid molecules may be introduced into mammalian cells by viral vectors. Methods of transforming cells are well known in the art. See, for example, U.S. Patent Nos. 4,399,216; 4,912,040; 4,740,461 and 4,959,455.

[0277] Described herein are recombinant methods for making an antibody or antigen-binding fragment thereof of the present disclosure, or an immunoglobulin chain thereof, comprising introducing a polynucleotide encoding one or more immunoglobulin chains of the antibody or fragment thereof (e.g., heavy and / or light immunoglobulin chain); culturing the host cell (e.g., CHO or Pichia (e.g.. Pichia pastoris)) under condition favorable to such expression and. optionally, isolating the antibody or fragment thereof or chain from the host cell and / or medium in which the host cell is grown. Antibodies can also be synthesized by any of the methods set forth in U.S. Patent No. 6,331,415.

[0278] Eukaryotic and prokaryotic host cells, including mammalian cells as hosts for expression of the antibodies or fragments thereof or immunoglobulin chains disclosed herein are well known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC). These include, inter alia, Chinese hamster ovary (CHO) cells. NSO, SP2 cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, 3T3 cells, HEK-293 cells and a number of other cell lines. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, bovine, horse and hamster cells. Cell lines may be selected by determining which cell lines have high expression levels. Other cell lines that may be used are insect cell lines, such as Sf9 cells, amphibian cells, bacterial cells, plant cells and fungal cells. Fungal cells include yeast and filamentous fungus cells including, for example, Pichia sp. (e.g., Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens , Pichia minuta (Ogataea minuta, Pichia lindneri), Pichia opuntiae. Pichia thermotolerans , Pichia salictaria, Pichia guercuum, Pichia pijperi, Pichia stiptis, or Pichia methanolica). Saccharomyces sp. (e.g., Saccharomyces cerevisiae), Hansenula polymorpha, Kluyveromyces sp. (e.g., Kluyveromyces lactis), Candida sp. (e.g., Candida albicans)' , Aspergillus sp. (e.g., Aspergillus nidulans, Aspergillus niger, or Aspergillus oryzae), Trichoderma reesei, Chrysosporium lucknowense, Fusarium sp. (e.g.. Fusarium gramineum or Fusarium venenatum). Physcomitrella patens and Neurospora crassa.

[0279] When recombinant expression vector(s) encoding the heavy chain or antigen-binding portion or fragment thereof, the light chain or antigen-binding fragment thereof, or both, are introduced into host cells, the antibodies are produced by culturing the host cells for a period of time sufficient to allow for expression of the antibody or fragment thereof or chain in the host cells or secretion of the into the culture medium in which the host cells are grown.

[0280] Antibodies and antigen-binding fragments thereof and immunoglobulin chains can be recovered from the culture medium using standard protein purification methods. Further, expression of antibodies and antigen-binding fragments thereof and immunoglobulin chains (or other moieties therefrom) are described herein. The expression for examples is achieved from production cell lines that can be enhanced using a number of known techniques. For example, the glutamine synthetase gene expression system (the GS system) is a common approach for enhancing expression under certain conditions. The GS system is discussed in European Patent Nos. 0 216 846, 0 256 055, and 0 323 997 and European Patent Application No. 89303964.4. The mammalian host cells (e.g., CHO) may lack a glutamine synthetase gene and are grown in the absence of glutamine in the medium w herein, how ever, the polynucleotide encoding theimmunoglobulin chain comprises a glutamine synthetase gene which complements the lack of the gene in the host cell.

[0281] In general, glycoproteins produced in a particular cell line or transgenic animal will have a glycosylation pattern that is characteristic for glycoproteins produced in the cell line (e.g., CHO cells) or transgenic animal. Therefore, the particular glycosylation pattern of an antibody will depend on the particular cell line or transgenic animal used to produce the antibody. However, all antibodies encoded by the nucleic acid molecules provided herein, or comprising the amino acid sequences provided herein, are included in the present disclosure, independent of the glycosylation pattern the antibodies may have. Similarly, in particular embodiments, antibodies with a glycosylation pattern comprising only non-fucosylated N-glycans may be advantageous, because these antibodies have been shown to typically exhibit more potent efficacy than their fucosylated counterparts both in vitro and in vivo (See for example, Shinkawa et al.. J. Biol. Chem. 278: 3466-3473 (2003); U.S. Patent Nos. 6,946,292 and 7,214,775). These antibodies with non-fucosylated N-glycans are not likely to be immunogenic because their carbohydrate structures are a normal component of the population that exists in human serum IgG.

[0282] Bispecific antibodies can be produced by a variety of methods including fusion of hybridomas or linking of Fab’ fragments. See. e.g., Songsivilai. et al., (1990) Clin. Exp. Immunol. 79: 315-321, Kostelny, et al., (1992) J Immunol. 148: 1547- 1553. In addition, bispecific antibodies may be formed as “diabodies” (Holliger et al., (1993) PNAS USA 90:6444- 6448) or as “Janusins” (Traunecker et al., (1991) EMBO J. 10:3655-3659 and Traunecker, et al., (1992) Int. J. Cancer Suppl. 7:51-52).Antibody Engineering

[0283] Further included are embodiments in which the antibodies (e.g., anti-CD27 antibodies or anti-PD-1 antibodies) and antigen-binding fragments thereof are engineered antibodies.

[0284] In certain embodiments, the antibodies and antigen-binding fragments thereof are engineered to include modifications in the framework and / or CDRs to improve their properties. Such engineered changes can be based on molecular modeling. A molecular model for the variable region for the parental (non-human) antibody sequence can be constructed to understand the structural features of the antibody and used to identify potential regions on the antibody that can interact with the antigen. Conventional CDRs are based on alignment of immunoglobulin sequences and identifying variable regions. Kabat et al., (1991) Sequences of Proteins of Immunological Interest, Kabat, et al; National Institutes of Health, Bethesda, Md.; 5th ed.; NIHPubl. No. 91-3242: Kabat (1978) Adv. Prot. Chem. 32: 1-75; Kabat. el al., (1977) J. Biol. Chem. 252:6609-6616. Chothia and coworkers carefully examined conformations of the loops in crystal structures of antibodies and proposed hypervariable loops. Chothia, et al., (1987) J Mol. Biol. 196:901-917 or Chothia, et al., (1989) Nature 342:878-883. There are variations between regions classified as CDRs and hypervariable loops. Later studies (Raghunathan et al., (2012) J. Mol Recog. 25, 3, 103-113) analyzed several antibody -antigen crystal complexes and observed that the antigen binding regions in antibodies do not necessarily conform strictly to the CDR residues or hypervariable loops. The molecular model for the variable region of the non-human antibodycan be used to guide the selection of regions that can potentially bind to the antigen. In practice, the potential antigen binding regions based on model differ from the conventional CDRs or hypervariable loops. Commercial scientific software such as MOE (Chemical Computing Group) can be used for molecular modeling. Human frameworks can be selected based on best matches with the non-human sequence both in the frameworks and in the CDRs. For FR4 (framework 4) in VH, VJ regions for the human germlines are compared with the corresponding non-human region. In the case of FR4 (framework 4) in VL, J-kappa and J-Lambda regions of human germline sequences are compared with the corresponding non-human region. Once suitable human frameworks are identified, the CDRs are grafted into the selected human frameworks. In some cases, the certain residues in the VL-VH interface can be retained as in the non-human (parental) sequence. Molecular models can also be used for identifying residues that can potentially alter the CDR conformations and hence binding to antigen. In some cases, these residues are retained as in the non-human (parental) sequence. Molecular models can also be used to identify- solvent exposed amino acids that can result in unwanted effects such as glycosylation, deamidation and oxidation. Developability filters can be introduced early on in the design stage to eliminate / minimize these potential problems.

[0285] In some examples, the engineered antibodies include modifications to framework residues within the variable domains of the antibody, e.g., to improve the properties of the antibody or fragment thereof. Typically, such framework modifications are made to decrease the immunogenicity of the antibody or fragment thereof. This is usually accomplished by replacing non-CDR residues in the variable domains (i.e., framework residues) in a parental (e.g., rodent) antibody or fragment thereof with analogous residues from the immune repertoire of the species in which the antibody is to be used, e.g.. human residues in the case of human therapeutics. In some embodiments, it is desirable to increase the affinity, or alter the specificity of an engineered (e.g., humanized) antibody. One approach is to “backmutate” one or more framework residues to the corresponding germline sequence. More specifically, an antibody or fragment thereof that hasundergone somatic mutation can contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody or fragment framework sequences to the germline sequences from which the antibody or fragment thereof is derived. Another approach is to revert to the original parental (e.g., rodent) residue at one or more positions of the engineered (e.g.. humanized) antibody, e.g.. to restore binding affinity that may have been lost in the process of replacing the framework residues. (See, e.g., U.S. Patent No. 5,693,762, U.S. Patent No. 5,585,089 and U.S. Patent No. 5,530,101.)

[0286] Another type of modification involves mutating one or more residues within the framework region, or even within one or more CDR regions, to remove T cell epitopes to thereby reduce the potential immunogenicity’ of the antibody. This approach is also referred to as “deimmunization” and is described in further detail in U.S. Patent No. 7,125,689.

[0287] In some embodiments, engineering may include changing certain amino acids containing exposed side-chains to another amino acid residue in order to provide for greater chemical stability of the final antibody, so as to avoid deamidation or isomerization. The deamidation of asparagine may occur on NG, DG, NG, NS, NA, NT, QG or QS sequences and result in the creation of an isoaspartic acid residue that introduces a kink into the polypeptide chain and decreases its stability (isoaspartic acid effect). Isomerization can occur at DG, DS, DA or DT sequences. In certain embodiments, the antibodies do not contain deamidation or asparagine isomerism sites.

[0288] For example, an asparagine (Asn) residue may be changed to Gin or Ala to reduce the potential for formation of isoaspartate at any Asn-Gly sequences, particularly within a CDR.

[0289] A similar problem may occur at an Asp-Gly sequence. See, e.g.. Reissner and Aswad (2003) Cell. Mol. Life Sci. 60: 1281. Isoaspartate formation may debilitate or completely abrogate binding of an antibody to its target antigen. See, e g., Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0290] In one embodiment, the asparagine is changed to glutamine (Gin). It may also be desirable to alter an amino acid adjacent to an asparagine (Asn) or glutamine (Gin) residue to reduce the likelihood of deamidation, which occurs at greater rates when small amino acids occur adjacent to asparagine or glutamine. See, Bischoff & Kolbe (1994) J. Chromatog. 662:261. In addition, Met residues (typically solvent exposed Met) in CDRs may be changed to Lys, Leu, Ala, or Phe or other amino acids in order to reduce the possibility that the methionine sulfur would oxidize, which could reduce antigen-binding affinity and also contribute to molecular heterogeneity in the final antibody preparation. Additionally, in order to prevent or minimize potential scissile Asn-Pro peptide bonds, it may be desirable to alter any Asn-Pro combinationsfound in a CDR to Gin-Pro, Ala-Pro, or Asn-Ala. Antibodies with such substitutions are subsequently screened to ensure that the substitutions do not decrease the affinity or specificity of the antibody for CD27, or other desired biological activity to unacceptable levels. Exemplary variants for stabilizing CDRs are shown in Table 5.Table 5. Exemplary stabilizing CDR variantsAntibody Engineering of the Fc Region

[0291] The antibodies (e.g., anti-CD27 antibodies or anti-PD-1 antibodies) and antigen-binding fragments thereof disclosed herein can also be engineered to include modifications within the Fc region, typically to alter one or more properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or effector function (e.g., antigen-dependent cellular cytotoxicity). Furthermore, the antibodies and antigen-binding fragments thereof disclosed herein can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or be modified to alter its glycosylation, again to alter one or more properties of the antibody or fragment thereof. Each of these embodiments is described in further detail below. The numbering of residues in the Fc region is that of the EU index of Kabat. The antibodies and antigen-binding fragments thereof disclosed herein also include antibodies and fragments with modified (or blocked) Fc regions to provide altered effector functions. See. e.g., U.S. Pat. No. 5,624,821; and PCT publications numbers W02003 / 086310; W02005 / 120571; and W02006 / 0057702. Such modifications can be used to enhance or suppress various reactions of the immune system, with possible beneficial effects in diagnosis and therapy. Alterations of the Fc region include amino acid changes (substitutions, deletions, and insertions), glycosylation or deglycosylation, and adding multiple Fc regions. Changes to the Fc can also alter the half-life of antibodies in therapeutic antibodies, enabling less frequent dosing and thus increasedconvenience and decreased use of material. See Presta (2005) J. Allergy Clin. Immunol. 116:731 at 734-35.

[0292] In one embodiment, the antibody or antigen-binding fragment thereof described herein is an IgG4 isotype antibody or fragment thereof comprising a Serine to Proline mutation at a position corresponding to position 228 (S228P; EU index) in the hinge region of the heavy chain constant region. This mutation has been reported to abolish the heterogeneity of inter-heavy chain disulfide bridges in the hinge region (Angal et al. supra; position 241 is based on the Kabat numbering system).

[0293] In one embodiment of the invention, the hinge region of CHI is modified such that the number of cysteine residues in the hinge region is increased or decreased. This approach is described further in U.S. Patent No. 5,677,425. The number of cysteine residues in the hinge region of CHI is altered, for example, to facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.

[0294] In another embodiment, the Fc hinge region of an antibody or antigen-binding fragment thereof is mutated to decrease the biological half-life of the antibody or fragment thereof. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc- hinge fragment such that the antibody or fragment thereof has impaired Staphylococcal protein A (SpA) binding relative to native Fc-hinge domain SpA binding. This approach is described in further detail in U.S. Patent No. 6,165,745.

[0295] In another embodiment, the antibody or antigen-binding fragment thereof is modified to increase its biological half-life. Various approaches are possible. For example, one or more of the following mutations can be introduced: T252L, T254S, T256F, as described in U.S. Patent No. 6,277,375. Alternatively, to increase the biological half-life, the antibody can be altered within the CHI or CL region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG, as described in U.S. Patent Nos. 5,869,046 and 6,121,022.

[0296] In yet other embodiments, the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector function(s) of the antibody or antigen-binding fragment thereof. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320 and 322 can be replaced with a different amino acid residue such that the antibody has an altered affinity for an effector ligand and retains the antigen-binding ability of the parent antibody. The effector ligand to which affinity is altered can be, for example, an Fc receptor or the Cl component of complement. This approach is described in further detail in U.S. Patent Nos. 5,624,821 and 5,648,260.

[0297] In another example, one or more amino acids selected from amino acid residues 329. 331 and 322 can be replaced with a different amino acid residue such that the antibody has altered Clq binding and / or reduced or abolished complement dependent cytotoxicity (CDC). This approach is described in further detail in U.S. Patent No. 6,194,551.

[0298] In another example, one or more amino acid residues within amino acid positions 231 and 239 are altered to thereby alter the ability of the antibody to fix complement. This approach is described further in PCT Publication WO 94 / 29351.

[0299] In some embodiments, the Fc region of an antibody is modified to increase or reduce the ability of the antibody or antigen-binding fragment thereof to mediate effector function and / or to increase / decrease their binding to the Fc gamma receptors (FcyRs).

[0300] For example, in some embodiments, the Fc region is modified to decrease the ability of the antibody or antigen-binding fragment thereof to mediate antibody dependent cellular cytotoxicity’ (ADCC) and / or to decrease the affinity of the antibody or fragment thereof for an Fey receptor by modifying one or more amino acids at the following positions: 238, 239. 243, 248, 249, 252, 254, 255, 256, 258, 264, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285,286, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322, 324,326, 327. 329, 330, 331, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, 388, 389, 398,414, 416. 419, 430, 434, 435, 437. 438 or 439. This approach is described further in PCTPublication WO 00 / 42072. Moreover, the binding sites on human IgGl for FcyRl, FcyRII, FcyRIII and FcRn have been mapped and variants with improved binding have been described (see Shields et al. (2001) J. Biol. Chem. 276:6591-6604).

[0301] In one embodiment, the Fc region is modified to decrease the ability of the antibody to mediate effector function and / or to increase anti-inflammatory properties by modifying residues 243 and 264. In one embodiment, the Fc region of the antibody or fragment thereof is modified by changing the residues at positions 243 and 264 to alanine. In one embodiment, the Fc region is modified to decrease the ability of the antibody or fragment thereof to mediate effector function and / or to increase anti-inflammatory properties by modifying residues 243. 264, 267 and 328.

[0302] The interaction between the constant region of an antigen binding protein and various Fc receptors (FcR) including Fc gammaRI (CD64), Fc gammaRIl (CD32) and Fc gammaRIII (CD 16) is believed to mediate the effector functions, such as ADCC and CDC, of the antigen binding protein. The Fc receptor is also important for antibody cross-linking, which can be important for anti -tumor immunity’.

[0303] Effector function can be measured in a number of ways including for example via binding of the Fc gammaRIII to Natural Killer cells or via Fc gammaRI tomonocytes / macrophages to measure for ADCC effector function. For example, an antigen binding protein of the present invention can be assessed for ADCC effector function in a Natural Killer cell assay. Examples of such assays can be found in Shields et al., 2001 J. Biol. Chem., Vol. 276. p 6591-6604; Chappel et al.. 1993 J. Biol. Chem.. Vol 268, p 25124-25131; Lazar et al., 2006 PNAS, 103; 4005-4010.

[0304] Human IgGl constant regions containing specific mutations or altered glycosylation on residue Asn297 have been shown to reduce binding to Fc receptors. In other cases, mutations have also been shown to enhance ADCC and CDC (Lazar et al., PNAS 2006, 103; 4005-4010; Shields et al. J Biol Chem 2001, 276; 6591-6604; Nechansky et al.. Mol Immunol, 2007. 44; 1815-1817).

[0305] In one embodiment of the present disclosure, such mutations are in one or more of positions selected from 239, 332 and 330 (IgGl), or the equivalent positions in other IgG isotypes. Examples of suitable mutations are S239D and I332E and A330L. In one embodiment, the antigen binding protein of the invention herein described is mutated at positions 239 and 332, for example S239D and I332E or in a further embodiment it is mutated at three or more positions selected from 239 and 332 and 330, for example S239D and I332E and A330L (EU index numbering).

[0306] In an alternative embodiment, provided herein is an antibody or antigen binding fragment thereof comprising a heavy chain constant region with an altered glycosylation profile such that the antigen binding protein has enhanced effector function. For example, wherein the antibody has enhanced ADCC or enhanced CDC or wherein it has both enhanced ADCC and CDC effector function. Examples of suitable methodologies to produce antigen binding proteins with an altered glycosylation profile are described in PCT Publication Nos. W02003011878 and W02006014679 and European Patent Application No. EP1229125.

[0307] In a further aspect, the present disclosure provides “non-fucosylated” or “afucosylated’' antibodies. Non-fucosylated antibodies harbor a tri-mannosyl core structure of complex-type N- glycans of Fc without fucose residue. These glycoengineered antibodies that lack core fucose residue from the Fc N-glycans may exhibit stronger ADCC than fucosy lated equivalents due to enhancement of Fc gammaRIIIa binding capacity.

[0308] Methods for producing an antibody or antigen binding fragment thereof may include the steps of a) culturing a recombinant host cell comprising an expression vector comprising an isolated nucleic acid as described herein, wherein the recombinant host cell does not comprise an alpha-1, 6-fucosyltransferase; and b) recovering the antigen binding protein. The recombinant host cell may not normally contain a gene encoding an alpha-1, -fucosyltransferase (for exampleyeast host cells such as Pichia sp.) or may have been genetically modified to inactivate an alpha- 1,6-fucosyltransferase. Recombinant host cells which have been genetically modified to inactivate the FUT8 gene encoding an alpha- 1,6-fucosyltransferase are available. See, e.g., the POTELLIGENT™ technology system available from BioWa, Inc. (Princeton, N.J.) in which CHOK1SV cells lacking a functional copy of the FUT8 gene produce monoclonal antibodies having enhanced ADCC activity that is increased relative to an identical monoclonal antibody produced in a cell with a functional FUT8 gene. Aspects of the POTELLIGENT™ technology system are described in US7214775. US6946292, W00061739 and W00231240. Those of ordinary skill in the art will also recognize other appropriate systems.

[0309] It will be apparent to those skilled in the art that such modifications may not only be used alone but may be used in combination with each other in order to further enhance or decrease effector function.Production of Antibodies with Modified Glycosylation

[0310] In still another embodiment, the antibodies (e g., anti-CD27 antibodies or anti-PD-1 antibodies) or antigen-binding fragments comprise a particular glycosylation pattern. For example, an afucosylated or an aglycosylated antibody or fragment thereof can be made (i.e., the antibody lacks fucose or glycosylation, respectively). The glycosylation pattern of an antibody or fragment thereof may be altered to, for example, increase the affinity or avidity’ of the antibody or fragment thereof for an antigen. Such modifications can be accomplished by, for example, altering one or more of the glycosylation sites within the antibody or fragment thereof sequence. For example, one or more amino acid substitutions can be made that result in removal of one or more of the variable region framework glycosylation sites to thereby eliminate glycosylation at that site. Such aglycosylation may increase the affinity or avidity of the antibody or fragment thereof for antigen. See, e.g., U.S. Patent Nos. 5,714,350 and 6,350,861.

[0311] Antibodies and antigen-binding fragments may further include those produced in lower eukaryote host cells, in particular fungal host cells such as yeast and filamentous fungi that have been genetically engineered to produce glycoproteins that have mammalian- or human-like glycosylation patterns (See for example, Choi et al., (2003) Proc. Natl. Acad. Sci. 100: 5022- 5027; Hamilton et al., (2003) Science 301 : 1244-1246; Hamilton et al., (2006) Science 313: 1441-1443; Nett et al., Yeast 28(3):237-52 (2011); Hamilton et al., Curr. Opin. Biotechnol. Oct;18(5):387-92 (2007)). A particular advantage of these genetically modified host cells over currently used mammalian cell lines is the ability to control the glycosylation profile of glycoproteins that are produced in the cells such that compositions of glycoproteins can beproduced wherein a particular N-glycan structure predominates (see, e.g., U.S. Patent No. 7,029,872 and U.S. Patent No. 7,449,308). These genetically modified host cells have been used to produce antibodies that have predominantly particular N-glycan structures (See for example, Li et al., (2006) Nat. Biotechnol. 24: 210-215).

[0312] In particular embodiments, the antibodies and antigen-binding fragments thereof further include those produced in lower eukaryotic host cells and which comprise fucosylated and non- fucosylated hybrid and complex N-glycans, including bisected and multiantennary species, including but not limited to N-glycans such as GlcNAc(i-4)Man3GlcNAc2;Gal(i-4)GlcNAc(i- 4>Man3GlcN Ac2i or NANA(i-4)Gal(i-4)GlcNAc(i-4)Man3GlcNAc2.

[0313] In particular embodiments, the antibodies and antigen-binding fragments thereof may comprise antibodies or fragments having at least one hybrid N-glycan selected from the group consisting of GlcNAcMansGlcNAc2; GalGlcN AcMansGlcN Ac2; and NANAGalGlcNAcMansGlcNAc2. In particular aspects, the hybrid N-glycan is the predominant N-glycan species in the composition.

[0314] In particular embodiments, the antibodies and antigen-binding fragments thereof comprise antibodies and fragments having at least one complex N-glycan selected from the group consisting of GlcNAcMan3GlcNAc2; GalGlcNAcMan3GlcNAc2;N ANAGalGlcN AcMamGlcN Ac2i GlcNAc2Man3GlcNAc2; GalGlcNAc2Man3GlcNAc2; Gal2GlcNAc2MansGlcNAc2; NANAGal2GlcNAc2Man3GlcNAc2; and NANA2Gal2GlcNAc2Man3GlcNAc2. In particular aspects, the complex N-glycan are the predominant N-glycan species in the composition. In further aspects, the complex N-glycan is a particular N-glycan species that comprises about 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, or 100% of the complex N- glycans in the composition. In one embodiment, the antibody and antigen binding fragments thereof provided herein comprise complex N-glycans, wherein at least 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, or 100% of the complex N- glycans comprise the structure NANA2Gal2GlcNAc2Man3GlcNAc2, wherein such structure is afucosylated. Such structures can be produced, e.g., in engineered Pichia pastoris host cells.

[0315] In some embodiments, the N-glycan is fucosylated. In general, the fucose is in an al,3-linkage with the GlcNAc at the reducing end of the N-glycan, an al,6-linkage with the GlcNAc at the reducing end of the N-glycan, an al,2-linkage with the Gal at the non- reducing end of the N-glycan. an al,3-linkage with the GlcNac at the non-reducing end of the N-glycan, or an al ,4-linkage with a GlcNAc at the non-reducing end of the N-glycan.

[0316] Therefore, in some aspects of the above the glycoprotein compositions, the glycoform is in an al,3-linkage or al ,6-linkage fucose to produce a glycoform selected from the groupconsisting of MansGlcNAc2(Fuc). GlcNAcMan5GlcNAc2(Fuc). Man3GlcNAc2(Fuc). GlcNAcMansGlcNAc2(Fuc), GlcN Ac2MansGlcNAc2(Fuc), GalGlcNAc2Man3GlcNAc2(Fuc), GahGlcNAc2Man3GlcNAc2(Fuc), NANAGal2GlcNAc2MansGlcNAc2(Fuc), and NANA2Gal2GlcNAc2Man3GlcNAc2(Fuc); in an al,3-linkage or al,4-linkage fucose to produce a glycoform selected from thegroup consisting of GlcNAc(Fuc)Man5GlcNAc2, GlcNAc(Fuc)Man3GlcNAc2, G1CNAC2(FUCI- 2)Man3GlcNAc2, GalGlcNAc2(Fuci-2)Man3GlcNAc2, Gal2GlcNAc2(Fuci-2)Man3GlcNAc2, NANAGal2GlcNAc2(Fuci-2)Man3GlcNAc2, and NANA2Gal2GlcNAc2(Fuci-2)Man3GlcNAc2; or in an al,2-linkage fucose to produce a glycoform selected from the group consisting of Gal(Fuc)GlcNAc2Man3GlcNAc2, Gal2(Fuci-2)GlcNAc2Man3GlcNAc2, NANAGal2(Fuci-2)GlcNAc2Man3GlcNAc2, and NANA2Gal2(Fuci-2)GlcNAc2Man GlcNAc2.

[0317] In further examples, the antibodies (e.g., humanized antibodies) or antigen-binding fragments thereof may comprise high mannose N-glycans, including but not limited to, MansGlcNAc2, Man?GlcNAc2, ManeGlcNAc2, MansGlcNAc2, Man4GlcNAc2, or N-glycans that consist of the Man3GlcNAc2N-glycan structure.

[0318] In further aspects of the above, the complex N-glycans may further include fucosylated and non-fucosylated bisected and multiantennary species.Antibody Physical Properties

[0319] The antibodies and antigen-binding fragments thereof disclosed herein may further contain one or more glycosylation sites in either the light or heavy chain immunoglobulin variable region. Such glycosylation sites may result in increased immunogenicity of the antibody or fragment thereof or an alteration of the pK of the antibody due to altered antigen-binding (Marshall et al. (1972) Annu Rev Biochem 41:673-702; Gala and Morrison (2004) J Immunol 172:5489-94; Wallick et al. (1988) J Exp Med 168: 1099-109; Spiro (2002) Glycobiology 12:43R-56R; Parekh et al. (1985) Nature 316:452-7; Mimura et al. (2000) Mol Immunol 37:697- 706). Glycosylation has been known to occur at motifs containing an N-X-S / T sequence.

[0320] Each antibody or antigen-binding fragment thereof has a unique isoelectric point (pl), which generally falls in the pH range between 6 and 9.5. The pl for an IgGl antibody typically falls within the pH range of 7-9.5 and the pl for an IgG4 antibody typically falls within the pH range of 6-8.

[0321] Each antibody or antigen-binding fragment thereof has a characteristic melting temperature, with a higher melting temperature indicating greater overall stability' in vivo(Krishnamurthy R and Manning MC 2002 Curr Pharm Biotechnol 3:361-71). In general, the TMi(the temperature of initial unfolding) may be greater than 60 °C, greater than 65 °C, or greater than 70 °C. The melting point of an antibody or fragment thereof can be measured using differential scanning calorimetry' (Chen et al., 2003 Pharm Res 20: 1952-60; Ghirlando et al., 1999 Immunol Lett 68:47-52) or circular dichroism (Murray et al., 2002 J. Chromatogr Sci 40:343-9).

[0322] In a further example, the antibodies and antigen-binding fragments thereof do not degrade rapidly. Degradation of an antibody or fragment thereof can be measured using capillary electrophoresis (CE) and MALDI-MS (Alexander AJ and Hughes DE (1995) Anal Chem 67:3626-32).

[0323] In a further example, the antibodies and antigen-binding fragments thereof have minimal aggregation effects, which can lead to the triggering of an unwanted immune response and / or altered or unfavorable pharmacokinetic properties. Generally, antibodies and fragments are acceptable with aggregation of 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less. Aggregation can be measured by several techniques, including size-exclusion column (SEC), high performance liquid chromatography (HPLC), and light scattering.Antibody Conjugates

[0324] The antibodies (e.g., anti-CD27 antibodies or anti-PD-1 antibodies) and antigen-binding fragments thereof disclosed herein may be conjugated to a chemical moiety'. The chemical moietys may be, inter alia, a polymer, a radionucleotide or a cytotoxic factor. In some embodiments, the chemical moiety' is a polymer which increases the half-life of the antibody or fragment thereof in the body of a subject. Suitable polymers include, but are not limited to, hydrophilic polymers which include but are not limited to polyethylene glycol (PEG) (e.g., PEG with a molecular weight of 2kDa, 5 kDa, 10 kDa, 12kDa, 20 kDa, 30kDa or 40kDa), dextran and monomethoxypolyethylene glycol (mPEG). Lee, et al., (1999) (Bioconj. Chem. 10:973-981) discloses PEG conjugated single-chain antibodies. Wen, et al., (2001) (Bioconj. Chem. 12:545- 553) disclose conjugating antibodies with PEG which is attached to a radiometal chelator (diethylenetriaminpentaacetic acid (DTP A)).

[0325] The antibodies and antigen-binding fragments disclosed herein may be PEGylated, for example to increase its biological (e.g., serum) half-life. To PEGylate an antibody or fragment thereof, the antibody or fragment thereof, typically is reacted with a reactive form of PEG, such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody' fragment thereof. In some embodiments, the PEGylation is carried out via an acylation reaction or an alkylation reaction with a reactive PEGmolecule (or an analogous reactive water-soluble polymer). In certain embodiments, the antibody or fragment thereof to be PEGylated is an aglycosylated antibody or fragment thereof. Methods for PEGylating proteins are known in the art and can be applied to the antibodies of the invention. See, e g., EP 0 154 316 and EP 0 401 384.

[0326] The antibodies and antigen-binding fragments thereof disclosed herein may be conjugated with labels such as "Tc,90Y,inIn,32P,14C,1251,3H,131I,nC,15O,13N,18F,35S,51Cr,57TO,226Ra,60Co,59Fe,57Se,152Eu,67CU,217Ci,21 1At.212Pb,47Sc,109Pd,234Th,40K,157Gd,55Mn,52Tr, and / or56Fe.

[0327] The antibodies and antigen-binding fragments disclosed herein may be conjugated with fluorescent or chemilluminescent labels, including fluorophores such as rare earth chelates, fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanate, phycoerythrin, phycocyanin, allophycocyanin, o-phthaladehyde, fluorescamine,152Eu, dansyl, umbelliferone, luciferin, luminal label, isoluminal label, an aromatic acridinium ester label, an imidazole label, an acndimium salt label, an oxalate ester label, an aequorin label. 2,3-dihydrophthalazinediones. biotin / avidin, spin labels, and stable free radicals.

[0328] The antibodies and antigen-binding fragments thereof also be conjugated to a cytotoxic factor such as diptheria toxin, Pseudomonas aeruginosa exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites for dii proteins and compounds (e.g., fatty acids), dianthin proteins, Phytoiacca americana proteins PAPI, PAPII, and PAP-S, Momordica charantia inhibitor, curcin, crotin, Saponaria officinalis inhibitor, mitogellin, restrictocin, phenomycin, and enomycin.

[0329] Any method known in the art for conjugating the antibodies and antigen-binding fragments thereof of the invention to the various moieties may be employed, including those methods described by Hunter, et al., (1962) Nature 144:945; David, et al., (1974) Biochemistry 13:1014; Pain, et al., (1981) J. Immunol. Meth. 40:219; and Nygren, J., (1982) Histochem. and Cytochem. 30:407.Pharmaceutical Compositions and Administration

[0330] To prepare pharmaceutical or sterile compositions of the compositions disclosed herein (e.g., anti-CD27 antibodies and antigen-binding fragments thereof, anti-PD-1 antibodies and antigen-binding fragments thereof, or chemotherapeutic agents), the composition may be admixed with a pharmaceutically acceptable carrier or excipient. See, e.g., Remington's Pharmaceutical Sciences and U.S. Pharmacopeia: National Formulary, Mack Publishing Company, Easton, PA (1984).

[0331] Formulations of therapeutic and diagnostic agents may be prepared by mixing with acceptable carriers, excipients, or stabilizers in the form of, e.g., lyophilized powders, slurries, aqueous solutions or suspensions (see, e.g., Hardman, et al. (2001) Goodman and Gilman’s The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy. Lippincott. Williams, and Wilkins, New York, NY; Avis, et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safely. Marcel Dekker, Inc., New York, NY).

[0332] Toxicity and therapeutic efficacy of the therapeutic agents disclosed herein, administered alone or in combination with another therapeutic agent, can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index (LD50 / ED50). The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration.

[0333] Any suitable route of administration may be used. Routes of administration (e.g., for an anti-CD27 antibody or antigen binding fragment thereof and / or an anti-PD-1 antibody or antigen binding fragment thereof) include oral, rectal, transmucosal, intestinal, parenteral; intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intratumor, intraocular, inhalation, insufflation, topical, cutaneous, transdermal, or intra-arterial. For example, the antibody or antigen binding fragment thereof may be administered intravenously or subcutaneously. In one example, the anti-CD27 antibody or antigen binding fragment thereof is administered intravenously by infusion. The administration may be systemic or local.

[0334] A vessel (e.g., a plastic or glass vial, a hollow bore needle, or a syringe cylinder) may be used that contains any of the therapeutic agents disclosed herein (e.g., an anti-CD27 antibody or antigen binding fragment thereof, an anti-PD- 1 antibody or antigen binding fragment thereof, and / or one or more chemotherapeutic agents) or a pharmaceutical composition thereof.

[0335] The present disclosure also provides an injection device comprising any of the therapeutic agents disclosed herein (e.g., an anti-CD27 antibody or antigen binding fragment thereof, an anti-PD-1 antibody or antigen binding fragment thereof, and / or one or more chemotherapeutic agents) or a pharmaceutical composition thereof. An injection device is a device that introduces a substance into the body of a patient via a parenteral route, e.g., intramuscular, subcutaneous or intravenous. For example, an injection device may be a syringe (e.g., pre-filled with the pharmaceutical composition, such as an auto-injector) which, for example, includes a cylinder or barrel for holding fluid to be injected (e.g., antibody or fragment thereof or a pharmaceutical composition thereof), a needle for piecing skin and / or blood vessels for injection of the fluid; and a plunger for pushing the fluid out of the cylinder and through the needle bore. In an embodiment, an injection device that comprises an antibody or antigenbinding fragment thereof of the present disclosure or a pharmaceutical composition thereof is an intravenous (IV) injection device. Such a device includes the antibody or fragment thereof or a pharmaceutical composition thereof in a cannula or trocar / needle which may be attached to a tube which may be attached to a bag or reservoir for holding fluid (e.g., saline; or lactated ringer solution comprising NaCl, sodium lactate, KC1, CaCh and optionally including glucose) introduced into the body of the patient through the cannula or trocar / needle. The antibody or fragment thereof or a pharmaceutical composition thereof may be introduced into the device once the trocar and cannula are inserted into the vein of a subject and the trocar is removed from the inserted cannula. The IV device may, for example, be inserted into a peripheral vein (e.g., in the hand or arm); the superior vena cava or inferior vena cava, or within the right atrium of the heart (e.g., a central IV); or into a subclavian, internal jugular, or a femoral vein and. for example, advanced toward the heart until it reaches the superior vena cava or right atrium (e.g., a central venous line). In an embodiment, an injection device is an autoinjector; ajet injector or an external infusion pump. Ajet injector uses a high-pressure narrow jet of liquid which penetrate the epidermis to introduce the antibody or fragment thereof or a pharmaceutical composition thereof to a patient's body. External infusion pumps are medical devices that deliver the antibody or fragment thereof or a pharmaceutical composition thereof into a patient’s body in controlled amounts. External infusion pumps may be powered electrically or mechanically. Different pumps operate in different ways, for example, a syringe pump holds fluid in the reservoir of a syringe, and a moveable piston controls fluid delivery, an elastomeric pump holds fluid in a stretchable balloon reservoir, and pressure from the elastic walls of the balloon drives fluid delivery. In a peristaltic pump, a set of rollers pinches down on a length of flexible tubing, pushing fluidforward. In a multi-channel pump, fluids can be delivered from multiple reservoirs at multiple rates.

[0336] The pharmaceutical compositions disclosed herein may also be administered with a needleless hypodermic injection device; such as the devices disclosed in U.S. Patent Nos. 6,620,135; 6.096,002; 5.399,163; 5,383.851; 5,312.335; 5,064,413; 4,941,880; 4,790,824 or 4,596,556. Such needleless devices comprising the pharmaceutical composition are also part of the present disclosure. The pharmaceutical compositions disclosed herein may also be administered by infusion. Examples of well-known implants and modules for administering the pharmaceutical compositions include those disclosed in: U.S. Patent No. 4.487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Patent No. 4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow implantable infusion apparatus for continuous drug delivery; U.S. Patent. No. 4,439,196. which discloses an osmotic drug delivery system having multi-chamber compartments. Many other such implants, delivery systems, and modules are well known to those skilled in the art and those comprising the pharmaceutical compositions of the present invention are within the scope of the present invention.

[0337] Alternately, one may administer the therapeutic agent (e.g., an anti-CD27 antibody or antigen binding fragment thereof, an anti-PD- 1 antibody or antigen binding fragment thereof, and / or one or more chemotherapeutic agents) in a local rather than systemic manner, for example, via injection of the antibody or fragment thereof directly into a tumor, e.g., a CD27+ tumor. Furthermore, one may administer the antibody or fragment thereof in a targeted drug delivery system, for example, in a hposome coated with a tissue-specific antibody, targeting, for example, a tumor e.g., a CD27+ tumor, e.g., characterized by immunopathology. The liposomes will be targeted to and taken up selectively by the afflicted tissue. Such methods and liposomes are part of the present disclosure.

[0338] With respect to antibodies or antigen-binding fragments thereof, the administration regimen may depend on several factors, including the serum or tissue turnover rate of the therapeutic antibody or antigen-binding fragment thereof, the level of symptoms, the immunogenicity of the therapeutic antibody, and the accessibility of the target cells in the biological matrix. Preferably, the administration regimen delivers sufficient therapeutic antibody or antigen-binding fragment thereof to effect improvement in the target disease state, while simultaneously minimizing undesired side effects. Accordingly, the amount of biologic delivered depends in part on the particular therapeutic antibody and the severity of the condition beingtreated. Guidance in selecting appropriate doses of therapeutic antibodies or fragments is available (see, e.g., Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert, et al. (2003) New Engl. J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med. 341: 1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343: 1594-1602). In various embodiments, the patient is provided a short, repeated exposure of the antibody or antigen binding fragment thereof rather than continuous exposure. Alternatively, the patient is provided a continuous exposure.

[0339] Antibodies or antigen-binding fragments thereof disclosed herein may be provided by continuous infusion, or by doses administered, e.g., daily. 1-7 times per week, weekly, bi-weekly, every three weeks, monthly, every five weeks, every six weeks, every seven weeks, bimonthly, quarterly, semiannually, or annually. Doses may be provided, e g., intravenously, subcutaneously, topically, orally, nasally, rectally, intramuscular, intracerebrally, intraspinally, or by inhalation, (see, e.g., Yang, et al., 2003 New Engl. J. Med. 349:427-434; Herold, et al., 2002 New Engl. J. Med. 346: 1692-1698; Liu. et al.. 1999 J. Neurol. Neurosurg. Psych. 67:451-456; Portielji, et al., 2003 Cancer Immunol. Immunother. 52: 151-144). Doses may also be provided to achieve a pre-determined target concentration of anti-CD27 antibody in the subject’s serum.

[0340] Also provided herein is a pharmaceutical composition for treating a cancer in a patient, the pharmaceutical composition comprising an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof, wherein the boserolimab is for use in combination with an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof and / or one or more chemotherapeutic agents (e.g., nab-paclitaxel), wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0341] Also provided herein is a pharmaceutical composition for treating a cancer in a patient, the pharmaceutical composition comprising an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof, wherein the anti-PD- 1 antibody or the antigen-binding fragment thereof is for use in combination with an anti-CD27 antibody (e.g., boserolimab) or an antigen-binding fragment thereof and / or one or more chemotherapeutic agents (e.g., nab- paclitaxel), wherein a tumor sample obtained from the patient has been determined to have a PD- L1 CPS of < 10.

[0342] Also provided herein is a pharmaceutical composition for treating a cancer in a patient, the pharmaceutical composition comprising one or more chemotherapeutic agents (e.g., nab- paclitaxel), wherein one or more chemotherapeutic agents is for use in combination with an anti- CD27 antibody (e.g.. boserolimab) or an antigen-binding fragment thereof and / or an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

[0343] Any of the therapeutic agents described herein, including anti-CD27 antibodies, anti- PD-1 antibodies, and / or chemotherapeutic agents disclosed herein, may be included in the pharmaceutical compositions described above.Kits

[0344] Further provided herein are kits comprising one or more components that include, but are not limited to, an anti-CD27 antibody or antigen-binding fragment thereof, as discussed herein (e.g., as described in Table 3) alone or in association w ith one or more additional components including, but not limited to a pharmaceutically acceptable carrier and / or one or more additional therapeutic agents, e.g., an anti-PD-1 antibody or antigen-binding fragment thereof or one or more chemotherapeutic agents. The anti-CD27 antibody or fragment thereof and / or the one or more additional therapeutic agents can be formulated as a pure composition or in combination with a pharmaceutically acceptable carrier, in a pharmaceutical composition.

[0345] In one embodiment, the kit includes an anti-CD27 antibody or antigen-binding fragment thereof or a pharmaceutical composition thereof in one container (e.g., in a sterile glass or plastic vial) and / or one or more additional therapeutic agents (e.g., an anti-PD-1 antibody or antigenbinding fragment thereof or one or more chemotherapeutic agents) or a pharmaceutical composition thereof in another container (e.g., in a sterile glass or plastic vial).

[0346] In another embodiment, the kit comprises an anti-CD27 antibody or antigen-binding fragment thereof of the invention along with a pharmaceutically acceptable carrier, optionally in combination with one or more therapeutic agents (e.g., an anti-PD-1 antibody or antigen- binding fragment thereof or one or more chemotherapeutic agents) formulated together, optionally, in a pharmaceutical composition, in a single, common container.

[0347] If the kit includes a pharmaceutical composition for parenteral administration to a subject, the kit can include a device for performing such administration. For example, the kit can include one or more hypodermic needles or other injection devices as discussed herein.

[0348] The kit can include a package insert including information concerning the pharmaceutical compositions and dosage forms in the kit. Generally, such information aidspatients and physicians in using the enclosed pharmaceutical compositions and dosage forms effectively and safely. For example, the following information regarding a therapeutic agent of the disclosure may be supplied in the insert: pharmacokinetics, pharmacodynamics, clinical studies, efficacy parameters, indications and usage, contraindications, warnings, precautions, adverse reactions, overdosage, proper dosage and administration, how supplied, proper storage conditions, references, manufacturer / distributor information and patent information.

[0349] Containers for use in such kits may typically comprise at least one vial, test tube, flask, bottle, syringe or other suitable container, into which one or more of the detection and / or therapeutic composition(s) may be placed, and preferably suitably aliquoted. Where one or more additional therapeutic agents is also provided, the kit may also contain one or more additional distinct containers into which the one or more additional therapeutic agents may be placed. Alternatively, a plurality of compounds may be prepared in a single pharmaceutical composition, and may be packaged in a single container means, such as a vial, flask, syringe, bottle, or other suitable single container. The kits disclosed herein will also typically include a means for containing the vial(s) in close confinement for commercial sale, such as, e g., injection or blow- molded plastic containers into which the desired vial(s) are retained.

[0350] Any of the kits provided herein may further include a diagnostic anti-PD-Ll antibody. Any suitable diagnostic anti-PD-Ll antibody may be included, such as 22C3 or 20C3.

[0351] Any of the kits disclosed herein may further include one or more reagents for a diagnostic assay, e.g., an IHC assay for detection of PD-L1. The reagent may be a detection reagent.

[0352] Any of the kits disclosed herein may further include a radiolabel, chromogenic, fluorogenic, or other A pe of detectable label or detecting means, which optionally may be conjugated to the diagnostic anti-PD-Ll antibody.

[0353] Where a radiolabel, chromogenic, fluorogenic, or other type of detectable label or detecting means is included within the kit. the labeling agent may be provided either in the same container as the detection or therapeutic composition itself or may alternatively be placed in a second distinct container means into which this second composition may be placed and suitably aliquoted. Alternatively, the detection reagent and the label may be prepared in a single container means, and in most cases, the kit will also typically include a means for containing the vial(s) in close confinement for commercial sale and / or convenient packaging and delivery.

[0354] A device or apparatus for carrying out the detection or monitonng methods described herein is also provided. Such an apparatus may include a chamber or tube into which sample can be input, a fluid handling system optionally including valves or pumps to direct flow of thesample through the device, optionally filters to separate plasma or serum from blood, mixing chambers for the addition of capture agents or detection reagents, and optionally a detection device for detecting the amount of detectable label bound to the capture agent immunocomplex. The flow of sample may be passive (e g., by capillary, hydrostatic, or other forces that do not require further manipulation of the device once sample is applied) or active (e.g.. by application of force generated via mechanical pumps, electroosmotic pumps, centrifugal force, or increased air pressure), or by a combination of active and passive forces.

[0355] In further embodiments, also provided is a processor, a computer readable memory, and a routine stored on the computer readable memory and adapted to be executed on the processor to perform any of the methods described herein. Examples of suitable computing systems, environments, and / or configurations include personal computers, server computers, hand-held or laptop devices, multiprocessor systems, microprocessor-based systems, set top boxes, programmable consumer electronics, network PCs, minicomputers, mainframe computers, distributed computing environments that include any of the above systems or devices, or any other systems known in the art.EXAMPLES

[0356] The following examples are meant to be illustrative and should not be construed as further limiting. The contents of the figures and all references, patents, and published patent applications cited throughout this application are expressly incorporated herein by reference.Example 1: Phase 1 evaluation of triple therapy with boserolimab (anti-CD27 agonist), pembrolizumab, and chemotherapy in patients with triple-negative breast cancer (TNBC) with PD-L1 combined positive score (CPS) <10

[0357] This Example describes a dose-expansion cohort of a first-in-human, open-label, phase 1 study (NCT03396445) for with escalating doses of the humanized anti-CD27 agonist antibody boserolimab.Obiectives / Hvpotheses and Endpoints

[0358] The study included male and female participants with advanced solid tumors who were at least 18 years of age. Table 6 summarizes objectives and endpoints of the study.Table 6. Objectives and EndpointsOverall Design

[0359] Table 7 summarizes the overall design of the study. See also FIG. 1.Table 7. Summary of Study DesignInclusion and Exclusion Criteria

[0360] Inclusion Criteria:• Arm 4: Triple-negative breast cancer (TNBC) that is locally recurrent, inoperable, not previously treated with chemotherapy, and which cannot be treated with curative intent OR metastatic disease not previously treated with chemotherapy.• Tumor PD-L1 CPS <10. To calculate PD-L1 CPS, the PD-L1 IHC 22C3 pharmDx assay was required for PD-L1 testing. Tissue samples could be tested locally to determine eligibility. If local testing was used, each site investigator ensured that PD-L1 testing was compliant with local regulations for guiding participant treatment decisions, notably that the PD-L1 IHC 22C3 pharmDx test kit w as used in accordance with instructions on the label. Tissue samples were also sent for central testing although participants could enter the study based on local PD-L1 IHC 22C3 pharmDx test results. When PD-L1 IHC 22C3 pharmDx testing could not be performed locally, central testing must have been performed to determine eligibility.• Measurable disease by RECIST 1.1, as assessed by the local site investigator / radiologist. Target lesions situated in a previously irradiated area were considered measurable if progression has been demonstrated in such lesions• Adequate organ function• Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1• Submit an evaluable baseline tumor sample for analysis (either a newly obtained or archival tumor sample)

[0361] Exclusion Criteria:• History of a second malignancy, unless potentially curative treatment has been completed with no evidence of malignancy for 2 years• Clinically active central nervous system metastases and / or carcinomatous meningitis• Has had a severe hypersensitivity reaction to treatment with a monoclonal antibody (mAb) and / or other components of the study treatment• Active infection requiring systemic treatment• History of interstitial lung disease• History of (noninfectious) pneumonitis that required steroids or current pneumonitis• Symptomatic ascites or pleural effusion• Previously had a stem cell or bone marrow transplant• Previously had a solid organ transplant• Active autoimmune disease that has required systemic treatment in the past 2 years (i.e., with use of disease-modifying agents, corticosteroids, or immunosuppressive drugs) except vitiligo or resolved childhood asthma / atopy• Known human immunodeficiency virus (HIV) and / or active and acute Hepatitis B or C infectionsNot fully recovered from any effects of major surgery without significant detectable infection• Pregnant or breastfeeding, or expecting to conceive or father children within the projected duration of the study• Had chemotherapy, definitive radiation, or biological cancer therapy within 4 weeks (2 weeks for palliative radiation) before the first dose of study treatment, or has not recovered to Grade <1 or better from any AEs that were due to cancer therapeutics administered more than 4 weeks earlier• Expected to require any other form of antineoplastic therapy while participating in this study• On chronic systemic steroid therapy in excess of replacement doses (e.g., exceeding 10 mg / day of prednisone equivalent), or on any other form of immunosuppressive medication• Regular user (including “recreational use”) of any illicit drugs at the time of signing informed consent, or has a recent history (within the last year) of substance abuse (including alcohol), as determined by the treating investigator. Participants who use cannabis for medicinal purposes or to treat specific symptoms will not be excluded unless it is being abused in the opinion of the treating investigator• Received a live-virus vaccine within 28 days before the first dose of study treatment• Currently participating and receiving study treatment in a study of an investigational agent or has participated and received study treatment in a study of an investigational agent or has used an investigational device within 28 days before the first dose of study treatment• Additional Exclusion Criteria for Participants in Arm 4:• Has a known history of hypersensitivity or allergy to nab-paclitaxel or any of its components• Has neuropathy >Grade 2• Has a history of class II-IV congestive heart failure or myocardial infarction within 6 months of randomization• Has received previous treatment with immune checkpoint inhibitor(s) (e.g., PD-l / PD- Ll)

[0362] Assessments:• PD-L1 expression in archival or newly obtained tumor tissue samples was assessed at a local or central laboratory using PD-L1 IHC 22C3 pharmDx• Adverse events (AEs) were assessed from the time of treatment allocation through 30 days after the last dose (90 days for serious AEs) and graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events version 4.03• Tumor imaging occurred at baseline and every 9 weeks after the first dose until disease progression, start of new anticancer treatment, pregnancy, withdrawal of consent, loss to followup, or death.• Baseline TcellinfGEP was assessed using the NanoString PanCancer Immune Panel• Baseline TMB was assessed using the Illumina TruSight™ Oncology 500 mutational panel assay

[0363] Analyses:• Safety analyses were performed in all patients who received >1 dose of study treatment• Antitumor activity was assessed in all treated patients who had measurable disease at baseline per RECIST version 1. 1 by investigator assessment• For ORR, point estimates and 95% Cis using the Clopper-Pearson CI method were provided• PFS was analyzed using the Kaplan-Meier method for censored data• Descriptive analyses for TcellinfGEP and TMB were conducted in treated patients who had corresponding biomarker data availableExample 2: Results from Phase 1 evaluation of triple therapy with boserolimab (anti-CD27 agonist), pembrolizumab, and chemotherapy in patients with triple-negative breast cancer (TNBC) with PD-L1 combined positive score (CPS) <10: Safety, antitumor activity, and association between biomarkers and responseBackground

[0364] Pembrolizumab plus chemotherapy is an approved therapy for patients with locally recurrent unresectable or metastatic TNBC whose tumors express PD-L1 (CPS >10) based on results from KEYNOTE-355. At the final analysis of the KEYNOTE-355 study, progression-free survival (PFS) results for pembrolizumab plus chemotherapy vs placebo plus chemotherapy were as follows: PD-L1 CPS >10: 9.7 vs 5.6 months (HR, 0.66; 95% CI, 0.50-0.88) PD-L1 CPS >1: 7.6 vs 5.6 months (HR, 0.75; 95% CI, 0.62-0.91) Intention-to-treat: 7.5 vs 5.6 months (HR, 0.82; 95% CI, 0.70-0.98). However, there is unmet need for effective treatment options in patients with PD-Ll CPS <10.

[0365] In preclinical studies, the combination of PD-1 blockade and antibody-based CD27 agonism improved antitumor activity and survival compared with CD27 agonism alone. In afirst-in-human, open -label, phase 1 study (NCT03396445), treatment with escalating doses of the humanized anti-CD27 agonist antibody boserolimab, alone and with pembrolizumab had acceptable safety and preliminary evidence of antitumor activity in patients with advanced solid tumors. Example 1 provides a summary of this study. This Example reports results for a doseexpansion cohort of this study in patients with metastatic TNBC and PD-L1 CPS <10 tumors who received first-line triple therapy with boserolimab, pembrolizumab, and chemotherapy. Exploratory analyses in this cohort assessed the association of baseline T-cell-inflamed gene expression profile (TcellintGEP) and tumor mutational burden (TMB) with ORR.Methods

[0366] This dose-expansion cohort enrolled patients aged >18 y with previously untreated locally recurrent inoperable or metastatic TNBC, measurable disease per RECIST vl . 1, Eastern Cooperative Oncology Group (ECOG) PS of 0 / 1, and tumors expressing PD-L1 at the cutoff of CPS <10 per PD-L1 IHC 22C3 pharmDx (local or central testing). Patients received boserolimab 30 mg intravenously (IV) every six weeks (Q6W) plus pembrolizumab 400 mg IV Q6W for 18 cycles plus nab-paclitaxel 100 mg / m2(3 weeks on days 1, 8, and 1 / 1 week off). Primary objectives were safety and tolerability7. A secondary objective was objective response rate (ORR) per RECIST vl. l by investigator assessment. Baseline TcellinfGEP was assessed using the NanoString PCI assay (N=31); TMB was assessed using the Illumina TSO500 mutational panel assay (N=21).Results

[0367] 41 patients were enrolled in the dose-expansion cohort. Median study follow -up at data cutoff was 11.1 (range, 2.2-20.5) months and median progression-free survival (PFS) was 10.3 (95% CI. 4.2-not reached) months. The most common treatment-related adverse events (AEs) were fatigue (51.2%), pruritus (51.2%), and alopecia (46.3%). Grade 3 / 4 treatment-related AEs occurred in 56.1% of patients (23 / 41); no grade 5 treatment-related AEs occurred. 9.8% of patients (4 / 41) discontinued >1 study treatment due to treatment-related AEs. Confirmed ORR was 46.3% (95% CI. 30.7%-62.6%), with 2 complete responses (CRs) and 17 partial responses (PRs); 14 patients had stable disease (SD), 5 had progressive disease (PD), and 3 had no postbaseline assessment at data cutoff. No clear association between TcellinfGEP or TMB and confirmed ORR was observed; responses were observed in patients with non-TcellintGEP-high (less than -0.318) and non-TMB-high (<10 mut / Mb) tumors.Conclusions

[0368] In patients with TNBC and tumors expressing PD-L1 at the cutoff of CPS <10, triple therapy with boserolimab, pembrolizumab, and chemotherapy had acceptable safety and showed evidence of antitumor activity regardless of baseline TcellinfGEP expression or TMB status.

[0369] Example 3: Additional results from Phase 1 evaluation of triple therapy with boserolimab, pembrolizumab, and chemotherapy in patients with TNBC with PD-L1 CPS <10: Safety, antitumor activity', and association between biomarkers and response This Example provides additional results from the phase 1 study described in Examples 1 and 2.Patients

[0370] 41 patients with IL metastatic TNBC expressing PD-L1 at the cutoff of CPS <10 were enrolled and treated (dose finding and dose expansion). 2 patients completed treatment, 26 discontinued treatment (clinical or radiographic progression, n = 20; AEs, n = 5; patient withdrawal, n = 1). and 13 were still on treatment at the data cutoff date. Median time from randomization to the data cutoff date was 21.4 (range, 12.9-27.9) months. Table 8 shows demographics and baseline characteristics.Table 8. Demographics and baseline characteristicsAll data are n (%) unless stated otherwise.Safety

[0371] Table 9 provides a summary of treatment related AEs. Table 10 provides a summary of immune-mediated AEs and infusion reactions.Table 9. Treatment- related AEsData are n (%) of patients.aPatients discontinued because of grade 3 infusion-related reactions (n = 2), grade 3 pneumonitis (n = 1). grade 2 peripheral sensory neuropathy (n = 1), grade 2 edema (n = 1), and grade 3 edema (n = l).Table 10. Immune-mediated AEs and infusion reactions*Data are n (%) of patients.aImmune-mediated AEs and infusion reactions were based on a list of preferred terms intended to capture known risks of pembrolizumab and were considered regardless of attribution to study treatment by the investigator.bPatients discontinued because of grade 3 infusion-related reactions (n = 2) and grade 3 pneumonitis (n = 1).Confirmed objective response

[0372] Table 11 provides a summary of antitumor activity and PFS per RECIST version 1.1 by investigator assessment.Table 11. Antitumor activity and PFS per RECIST version 1.1 by investigator assessmentCR, complete response; NR, not reached; PD, progressive disease; PR, partial response; SD, stable disease.aIncludes patients with confirmed CR or PR.bNo postbaseline assessment available for response evaluation.Biomarkers

[0373] A majority of responses were observed in tumors with negative PD-L1 (CPS <1) (FIG. 2). There was no clear association between TcellinfGEP or TMB and confirmed response (FIGS. 3A and 3B, respectively). Multiple confirmed responses were observed in patients with non- TcellinfGEP-high (less than -0.318) and TMB-low (<10 mut / Mb) tumors.Conclusions

[0374] Triple therapy with boserolimab, pembrolizumab, and chemotherapy had an acceptable safety profile in patients with TNBC and tumors of PD-L1 CPS <10. Evidence of antitumor activity was shown regardless of baseline PD-L1 status, TcellinfGEP expression, or TMB status.

[0375] The disclosed subject matter is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the disclosure in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0376] All references (e.g., publications or patents or patent applications) cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety' for all purposes. Other embodiments are within the following claims.

Claims

WHAT IS CLAIMED IS:

1. A method for treating a cancer in a patient, the method comprising administering to the patient a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof, wherein a tumor sample obtained from the patient has been determined to have a programmed death-ligand 1 (PD-L1) combined positive score (CPS) of less than (<) 10.

2. A method for treating a cancer in a patient, the method comprising: a) determining that a tumor sample obtained from the patient has a PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof to the patient.

3. The method of claim 1 or 2, wherein the cancer is triple-negative breast cancer (TNBC).

4. The method of claim 3, wherein the TNBC is (i) locally recurrent unresectable TNBC or (ii) metastatic TNBC.

5. The method of any one of claims 1-4, wherein the patient is previously untreated for the cancer.

6. The method of claim 5, wherein the patient has not been previously treated with a chemotherapy.

7. The method of any one of claims 1-6, wherein the PD-L1 CPS is determined using an immunohistochemistry (IHC) assay comprising the diagnostic anti-PD-Ll antibody 22C3.

8. The method of any one of claims 1-7, wherein the anti-CD27 antibody or the antigenbinding fragment thereof comprises: a) a heavy chain variable region complementarity determining region (CDR) 1 comprising the amino acid sequence of SEQ ID NO: 1 ; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO:2;c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ IDNO:3; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ IDNON; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO:5; and1) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

9. The method of any one of claims 1-8, wherein the anti-CD27 antibody or the antigenbinding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising SEQ ID NO:7 and the light chain comprises a light chain variable region comprising SEQ ID NO: 9.

10. The method of any one of claims 1-8, wherein the anti-CD27 antibody or the antigen binding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 8 and the light chain comprises the amino acid sequence of SEQ ID NOTO.

11. The method of any one of claims 1-10, wherein the anti-CD27 antibody is boserolimab.

12. The method of any one of claims 1-11, wherein the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion.

13. The method of any one of claims 1-12, wherein the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 30 mg.

14. The method of any one of claims 1-13, wherein the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient every three weeks (Q3W) or every six weeks (Q6W).

15. The method of any one of claims 1-14, wherein the anti-CD27 antibody or antigen binding fragment thereof is co-administered with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti-PD-Ll antibody or an antigen binding fragment thereof.

16. The method of any one of claims 1-14, wherein the anti-CD27 antibody or antigen binding fragment thereof is co-formulated with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti-PD-Ll antibody or an antigen binding fragment thereof.

17. The method of claim 16, wherein the anti-PD-1 antibody, or the antigen binding fragment thereof comprises: a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ IDNO: 16; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO: 17; c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ IDNO: 18; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ IDNO:11; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO: 12; and1) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 13.

18. The method of claim 16 or 17, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 14.

19. The method of any one of claims 15-17, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:20 and the light chain comprises the amino acid sequence of SEQ ID NO: 15.

20. The method of any one of claims 15-19, wherein the anti-PD-1 antibody is pembrolizumab.

21. The method of any one of claims 15-19, wherein the anti-PD-1 antibody is a pembrolizumab variant.

22. The method of any one of claims 15-21. wherein the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion.

23. The method of any one of claims 15-22, wherein the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 400 mg.

24. The method of any one of claims 15-23, wherein the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient every six weeks (Q6W).

25. The method of any one of claims 1-24, wherein the anti-CD27 antibody or antigen binding fragment thereof is co-administered with one or more chemotherapeutic agents.

26. The method of claim 25. wherein the one or more chemotherapeutic agents comprise nab- paclitaxel.

27. The method of claim 26, wherein the nab-paclitaxel is administered to the patient via intravenous infusion.

28. The method of claim 26 or 27, wherein the nab-paclitaxel is administered to the patient at a dose of about 100 mg / m2.

29. The method of any one of claims 26-28, wherein the nab-paclitaxel is administered to the patient on a 28-day cycle, and the nab-paclitaxel is administered to the patient on Day 1, 8, and 15 of the 28-day cycle.

30. The method of any one of claims 1-29, wherein the patient is greater than or equal to 18 years of age.

31. A method for treating a previously untreated locally recurrent unresectable TNBC or metastatic TNBC in a patient, the method comprising administering to the patient a therapeutically effective amount of boserolimab, pembrolizumab, and nab-paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

32. An anti-CD27 antibody or an antigen-binding fragment thereof for use in treating a cancer in a patient, wherein a tumor sample obtained from the patient has been determined to have a PD-Ll CPS of <10.

33. An anti-CD27 antibody or an antigen-binding fragment thereof for use in a method for treating a cancer in a patient, the method comprising: a) determining that a tumor sample obtained from the patient has a PD-L1 CPS of < 10; and b) administering a therapeutically effective amount of an anti-CD27 antibody or an antigen-binding fragment thereof to the patient.

34. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 32 or 33, wherein the cancer is triple-negative breast cancer (TNBC).

35. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 34, wherein the TNBC is (i) locally recurrent unresectable TNBC or (ii) metastatic TNBC.

36. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 32-35, wherein the patient is previously untreated for the cancer.

37. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 36, wherein the patient has not been previously treated with a chemotherapy.

38. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-37, wherein the PD-L1 CPS is determined using an IHC assay comprising the diagnostic anti-PD-Ll antibody 22C3.

39. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-38, wherein the anti-CD27 antibody or the antigen-binding fragment thereof comprises: a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ IDNO: 1; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO:2;c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ IDNO:3; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ IDNO:4; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO:5; and1) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

40. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-39, wherein the anti-CD27 antibody or the antigen-binding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising SEQ ID NO: 7 and the light chain comprises a light chain variable region comprising SEQ ID NO: 9.

41. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-39, wherein the anti-CD27 antibody or the antigen binding fragment thereof comprises a heavy chain and a light chain, and wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 8 and the light chain comprises the amino acid sequence of SEQ ID NOTO.

42. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-41, wherein the anti-CD27 antibody is boserolimab.

43. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-42, wherein the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion.

44. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-43, wherein the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 30 mg.

45. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-44, wherein the anti-CD27 antibody or antigen binding fragment thereof is administered to the patient every six weeks (Q6W).

46. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-45, wherein the anti-CD27 antibody or antigen binding fragment thereof is coadministered with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti- PD-L 1 antibody or an antigen binding fragment thereof.

47. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-45, wherein the anti-CD27 antibody or antigen binding fragment thereof is coformulated with (i) an anti-PD-1 antibody or an antigen binding fragment thereof or (ii) an anti- PD-L 1 antibody or an antigen binding fragment thereof.

48. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 47, wherein the anti-PD-1 antibody, or the antigen binding fragment thereof comprises: a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ IDNO: 16; b) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO: 17; c) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ IDNO: 18; d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ IDNO:11; e) a light chain variable region CDR2 comprising the amino acid sequence of SEQ IDNO: 12; andI) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 13.

49. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 47 or 48, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, and wherein the heavy chain comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and the light chain comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 14.

50. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 46-48, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, andwherein the heavy chain compnses the amino acid sequence of SEQ ID NO:20 and the light chain comprises the amino acid sequence of SEQ ID NO: 15.

51. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 46-50, wherein the anti-PD-1 antibody is pembrolizumab.

52. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 46-50, wherein the anti-PD-1 antibody is a pembrolizumab variant.

53. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 46-52, wherein the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient via intravenous infusion.

54. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 46-53, wherein the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient at a dose of about 400 mg.

55. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 46-54, wherein the anti-PD-1 antibody or antigen binding fragment thereof is administered to the patient every six weeks (Q6W).

56. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-55, wherein the anti-CD27 antibody or antigen binding fragment thereof is coadministered with one or more chemotherapeutic agents.

57. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 56. wherein the one or more chemotherapeutic agents comprise nab-paclitaxel.

58. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 57, wherein the nab-paclitaxel is administered to the patient via intravenous infusion.

59. The anti-CD27 antibody or the antigen-binding fragment thereof for use of claim 57 or 58, wherein the nab-paclitaxel is administered to the patient at a dose of about 100 mg / m260. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 57-59, wherein the nab-paclitaxel is administered to the patient on a 28-day cycle, and the nab-paclitaxel is administered to the patient on Day 1, 8, and 15 of the 28-day cycle.

61. The anti-CD27 antibody or the antigen-binding fragment thereof for use of any one of claims 33-60, wherein the patient is greater than or equal to 18 years of age.

62. Boserolimab for use in treating a previously untreated locally recurrent unresectable TNBC or metastatic TNBC in a patient, wherein the boserolimab is administered in combination with pembrolizumab and nab-paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

63. A pharmaceutical composition for treating a previously untreated locally recurrent unresectable TNBC or metastatic TNBC in a patient, the pharmaceutical composition comprising boserolimab, wherein the boserolimab is for use in combination with pembrolizumab and nab- paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD- L1 CPS of < 10.

64. A pharmaceutical composition for treating a previously untreated locally recunent unresectable TNBC or metastatic TNBC in a patient, the pharmaceutical composition comprising pembrolizumab, wherein the pembrolizumab is for use in combination with boserolimab and nab-paclitaxel, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

65. A pharmaceutical composition for treating a previously untreated locally recurrent unresectable TNBC or metastatic TNBC in a patient, the pharmaceutical composition comprising nab-paclitaxel. wherein the nab-paclitaxel is for use in combination with boserolimab and pembrolizumab, wherein a tumor sample obtained from the patient has been determined to have a PD-L1 CPS of < 10.

66. A kit comprising an anti-CD27 antibody or an antigen-binding fragment thereof, and instructions to administer the anti-CD27 antibody or the antigen-binding fragment thereof to a patient having a cancer, wherein a tumor sample obtained from the patient has been determined to have a PD-Ll CPS of <10.

67. The kit of claim 66, further comprising a diagnostic anti-PD-Ll antibody.

68. The kit of claim 67, wherein the diagnostic anti-PD-Ll antibody is 22C3.

69. The kit of any one of claims 66-68, further comprising one or more reagents for an IHC assay.

70. The kit of any one of claims 66-69, wherein the anti-CD27 antibody is boserolimab.

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