Molecules that bind to CD94 / NKG2A heterodimer polypeptides

Abstract

This document provides methods and materials involving binding of binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) to CD94 / NKG2A polypeptides. For example, provided are binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) that bind to CD94 / NKG2A polypeptides, as well as methods and materials for using one or more such binding molecules to treat a mammal (e.g., a human) with cancer and / or a viral infection.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 289,495, filed December 14, 2021. The disclosure of the prior application is considered part of (and is incorporated by reference into) the disclosure of this application.

[0002] 1.Technical Field This document relates to methods and materials involving binding of molecules (e.g., antibodies, antibody fragments, antibody domains, chimeric antigen receptors (CARs), cell engagers, or antibody drug conjugates (ADCs)) to CD94 / NKG2A heterodimeric polypeptides (referred to herein as CD94 / NKG2A polypeptides). For example, this document provides binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, or ADCs) that bind to CD94 / NKG2A polypeptides, and methods and materials of using such binding agents to treat cancer and / or viral infections (e.g., HIV infection). This document also provides cells (e.g., host cells) engineered to express one or more binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, or cell engagers) capable of binding to CD94 / NKG2A polypeptides, and methods and materials of using such cells to treat cancer and / or viral infections (e.g., HIV infection). [Background technology]

[0003] 2. Background information CD94 / NKG2A is an inhibitory C-type lectin heterodimeric receptor expressed on 20-60% of natural killer (NK) cells and specifically interacts with human leukocyte antigen (HLA) class I histocompatibility complex alpha chain E (HLA-E), also known as MHC class I antigen E. Ligation of CD94 / NKG2A to HLA-E mediates the phosphorylation of immunoreceptor tyrosine-based inhibitory motifs (ITIMs) on CD94 / NKG2A, resulting in the expression of NK cells and CD8 + As reported, upregulation of HLA-E expression in tumor cells and HIV-infected lymphocytes has been shown to enable tumor cells and HIV-infected lymphocytes to acquire the ability to escape elimination by immune cells through the induction of CD94 / NKG2A-mediated inhibitory signals. Summary of the Invention

[0004] overview This document provides methods and materials involving binding of molecules (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, or ADCs) to CD94 / NKG2A polypeptides. For example, this document provides binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, or ADCs) that bind to CD94 / NKG2A polypeptides, as well as methods and materials using one or more such binding agents to treat a mammal (e.g., a human) with cancer and / or a viral infection (e.g., HIV infection).

[0005] This document also provides cells (e.g., host cells) engineered to express one or more binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, or cell engagers) capable of binding to a CD94 / NKG2A polypeptide, as well as methods and materials for using such cells to treat cancer and / or viral infection (e.g., HIV infection).

[0006] As described herein, a binding agent (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more CARs, one or more cell engagers, and / or one or more ADCs) can be designed to have the ability to bind to a CD94 / NKG2A polypeptide. For example, a binding agent (e.g., an antibody, an antigen-binding fragment, an antibody domain, a CAR, a cell engager, or an ADC) provided herein can be capable of binding to a polypeptide comprising, consisting essentially of, or consisting of the amino acid sequence of a human CD94 / NKG2A polypeptide set forth in SEQ ID NO:74 and SEQ ID NO:75, SEQ ID NO:256 and SEQ ID NO:257, and / or SEQ ID NO:259 (see, e.g., FIG. 1).

[0007] In some cases, the binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) may be used to: (a) inhibit or inhibit NK cells or T cells (e.g., CD8 + (b) may, through said binding, block the ability of HLA-E present on cancer and / or virus-infected cells to trigger an inhibitory response in the NK cell or the T cell via the CD94 / NKG2A polypeptide, and / or (c) may, through said binding, cause internalization of the CD94 / NKG2A polypeptide, resulting in a decrease in surface expression of the CD94 / NKG2A polypeptide in the NK cell or the T cell. For example, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, cell engagers, or ADCs) can bind to CD94 / NKG2A polypeptides present on the surface of NK cells or T cells (e.g., CD8 +The NK cell or T cell may be capable of binding to a polypeptide comprising, consisting essentially of, or consisting of the amino acid sequence of a human CD94 / NKG2A polypeptide set forth in SEQ ID NO:74 and SEQ ID NO:75, SEQ ID NO:256 and SEQ ID NO:257, and / or SEQ ID NO:259 (see, e.g., FIG. 1 ) present on the surface of the NK cell or T cell, in a manner that does not elicit an inhibitory response in the NK cell or T cell, and in a manner that blocks the ability of HLA-E present on cancer and / or virus-infected cells to elicit an inhibitory response in the NK cell or T cell.

[0008] In some cases, two sets of three CDRs of the antigen-binding fragments provided herein (e.g., SEQ ID NOs: 1-3 and 9-11, or SEQ ID NOs: 17-19 and 25-27) are engineered into a CAR to confer a CD94 / NKG2A + CARs with cell targeting capabilities + Cells (e.g., CAR + T cells, CAR + CARs such as induced pluripotent stem cells + Stem Cells, or CARs + They can produce NK cells.

[0009] In some cases, two sets of three CDRs of the antigen-binding fragments provided herein (e.g., SEQ ID NOs: 1-3 and 9-11, or SEQ ID NOs: 17-19 and 25-27) are engineered into an antibody, antigen-binding fragment, or antibody domain structure to form a CD94 / NKG2A + NK cells and / or CD94 / NKG2A + CD8 + Binding agents (eg, fully human binding agents) can be made that have the ability to bind to CD94 / NKG2A polypeptides and inhibit their suppressive activity on T cells.

[0010] In some cases, two sets of three CDRs of the antigen-binding fragments provided herein (e.g., SEQ ID NOs: 1-3 and 9-11, or SEQ ID NOs: 17-19 and 25-27) can be engineered into a cell engager that targets an antigen of interest, such as a bispecific T cell engager (e.g., BiTE), trispecific T cell engager (e.g., TriTE), bispecific killer engager (e.g., BiKE), and / or trispecific killer engager (e.g., TriKE), to (a) bind to a CD94 / NKG2A polypeptide via a CD94 / NKG2A polypeptide. + Ability to bind to NK cells and / or CD94 / NKG2A via a CD94 / NKG2A polypeptide + T cells (e.g., CD94 / NKG2A + CD8 + In some cases, cell engagers provided herein (e.g., BiKE) may be generated that have the ability to bind to NK cells (e.g., NK and / or T cells) and (b) target such NK and / or T cells to cells expressing a target antigen of interest, thereby inducing one or more immune responses (e.g., NK and / or T cell immune responses and / or ADCC using a cell engager in the absence of an Fc-containing antibody) against the cells expressing the target antigen of interest. In some cases, cell engagers provided herein (e.g., BiKE) may bind to NK cells even though they do not contain an antigen-binding domain that binds CD16. In some cases, cell engagers provided herein (e.g., BiTE) may bind to T cells even though they do not contain an antigen-binding domain that binds CD3. Note that BiTE- and TriKE-mediated killing may be referred to as ADCC, even though it is not initiated by an Fc domain.

[0011] In addition, as described herein, a binding agent provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, and / or one or more antibody domains) may be used to generate a conjugate comprising the binding agent and a drug. For example, ADCs, such as whole antibody-drug conjugates, Fab-drug conjugates, and / or antibody domain drug conjugates, may be designed to include an appropriate binding agent provided herein to generate the conjugate. Such conjugates may be used to generate a conjugate comprising the binding agent and a drug. + NK cells and / or CD94 / NKG2A + T cells (e.g., CD94 / NKG2A + CD8 + The NK cell- and T cell-specific immunoglobulins may be used to deliver a payload to target cells, such as NK cells and / or T cells. In some cases, the payload may be a drug that promotes NK cell and / or T cell activity against cancer cells and / or virus-infected cells.

[0012] Also as described herein, the binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) may be used to treat a mammal (e.g., a human) with cancer. For example, a mammal (e.g., a human) with cancer may be treated with (a) an HLA-E polypeptide that inhibits the immune response of NK and / or T cells via the interaction of HLA-E with CD94 / NKG2A polypeptide present on the surface of the NK and / or T cells. + Compositions comprising one or more binding agents described herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) may be administered to block the ability of cancer cells, (b) to induce ADCC against cancer cells by targeting an antigen of interest expressed by cancer cells in the mammal, and / or (c) to increase the survival time of the mammal from cancer.

[0013] Also as described herein, the binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) may be used to treat a mammal (e.g., a human) having a viral infection (e.g., an HIV infection). For example, a mammal (e.g., a human) having a viral infection (e.g., an HIV infection) may be administered (a) an HLA-E polypeptide that inhibits the immune response of NK cells and / or T cells via the interaction of HLA-E with a CD94 / NKG2A polypeptide present on the surface of the NK cells and / or T cells. + Compositions comprising one or more binding agents described herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) may be administered to block the ability of virally infected cells, (b) to induce ADCC against virally infected cells by targeting an antigen of interest expressed by the virally infected cells in the mammal, and / or (c) to increase the survival time of the mammal from viral infection.

[0014] Also as described herein, a cell (e.g., a host cell) may be engineered to express one or more binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, or cell engagers) capable of binding to a CD94 / NKG2A polypeptide. For example, a cell such as a T cell (e.g., a CTL), a stem cell (e.g., an induced pluripotent stem cell), or an NK cell may be engineered to express one or more CARs capable of binding to a CD94 / NKG2A polypeptide. In some cases, such cells (e.g., CD94 / NKG2A-specific CARs) may be engineered to express one or more binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, or cell engagers) capable of binding to a CD94 / NKG2A polypeptide. + T cells or NK cells) may be used to modulate the tumor microenvironment (TME) by killing subsets of NK and / or T cells that secrete inhibitory cytokines and / or promote the inhibition of other immune cells.

[0015] In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) may be used to detect the presence or absence of a CD94 / NKG2A polypeptide. For example, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) may be used to detect the presence or absence of a CD94 / NKG2A polypeptide when a sample (e.g., a biological sample) obtained from a mammal (e.g., a human) detects CD94 / NKG2A + Cells (e.g., CD94 / NKG2A + NK cells and / or CD94 / NKG2A + T cells). + NK and / or CD94 / NKG2A + Having the ability to detect the presence or absence of CD94 / NKG2A T cells within a mammalian tumor site can help clinicians, medical professionals, and patients make better decisions regarding potential treatment options. + NK cells and / or CD94 / NKG2A + Detection of T cell abundance allows clinicians, healthcare professionals, and patients to better understand the CD94 / NKG2A + NK cells and / or CD94 / NKG2A + This allows the selection of appropriate anti-cancer treatments that promote T cell activity. CD94 / NKG2A + NK cells and / or CD94 / NKG2A + Such treatments to promote T cell activity can include, but are not limited to, administration of BiKE, TriTE, TriKE, and / or BiTE provided herein, which have the ability to bind to CD94 / NKG2A without inhibiting the NK cells or T cells and the ability to bind to a target antigen of interest expressed by cancer cells.

[0016] As described herein, the use of the binding agents described herein to block CD94 / NKG2A polypeptides from binding to HLA-E and / or from causing an inhibitory effect on NK cells and / or T cells can, for example, inhibit NK cell- and / or CD8 + Restoring T cell-mediated cytotoxic activity and / or enhancing the immunity of surrounding immune cells may provide an effective method for treating cancer and / or viral infections (e.g., HIV infection).

[0017] In general, one aspect of this document is a method for producing a light chain variable domain or region comprising: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with the addition, deletion or substitution of one, two or three amino acids), SEQ ID NO:2 (or SEQ ID NO:2 with the addition, deletion or substitution of one, two or three amino acids), and SEQ ID NO:3 (or SEQ ID NO:3 with the addition, deletion or substitution of one, two or three amino acids); and (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with the addition, deletion or substitution of one, two or three amino acids), SEQ ID NO:10 (or SEQ ID NO:10 with the addition, deletion or substitution of one, two or three amino acids), and SEQ ID NO:11 (or SEQ ID NO:11 with the addition, deletion or substitution of one, two or three amino acids); or In one embodiment, the present invention features an antibody comprising (or consisting essentially of, or consisting of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antibody may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16.The antibody may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 24. The antibody may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.

[0018] In another aspect, this document provides a method for detecting a mutated nucleotide sequence comprising: (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:17 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions). In one embodiment, the present invention features an antigen-binding fragment comprising (or consisting essentially of, or consisting of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antigen-binding fragment may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise the heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antigen-binding fragment may comprise the light chain variable domain or region of (i).The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16. The antigen-binding fragment may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 24. The antigen-binding fragment may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab.

[0019] In another aspect, this document features a chimeric antigen receptor comprising (or consisting essentially of, or consisting of) an antigen binding domain, a hinge, a transmembrane domain, and one or more signaling domains, wherein the antigen binding domain comprises an antibody or antigen binding fragment. The antibody comprises: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions). The antibody may comprise (or consist essentially of, or consist of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8. The antibody may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16. The antibody may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antibody may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody. The antigen-binding fragment comprises: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:17 with one, two or three amino acid additions, deletions or substitutions). , or may consist essentially of, or may consist of, a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17, or SEQ ID NO: 18 (or SEQ ID NO: 18 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 19 (or SEQ ID NO: 19 with 1, 2 or 3 amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with 1, 2 or 3 amino acid additions, deletions or substitutions), and SEQ ID NO: 27 (or SEQ ID NO: 27 with 1, 2 or 3 amino acid additions, deletions or substitutions). The antigen-binding fragment may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise the heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antigen-binding fragment may comprise the light chain variable domain or region of (i).The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16. The antigen-binding fragment may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 24. The antigen-binding fragment may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. The antigen-binding domain may comprise an scFv having the ability to bind to a CD94 / NKG2A polypeptide. The hinge may comprise a hinge as set forth in FIG. 13. The transmembrane domain may comprise a transmembrane domain as set forth in FIG. 14. The chimeric antigen receptor may comprise one or more signaling domains as set forth in FIG. 15.

[0020] In another aspect, this document features a cell that includes the chimeric antigen receptor of the preceding paragraph. The cell can be a T cell, a stem cell, or a NK cell.

[0021] In another aspect, this document features a cell engager comprising (or consisting essentially of, or consisting of) a first antigen-binding domain, a linker and a second antigen-binding domain, wherein the first antigen-binding domain comprises an antibody or antigen-binding fragment. The antibody comprises: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions). The antibody may comprise (or consist essentially of, or consist of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8. The antibody may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16. The antibody may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antibody may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.The antigen-binding fragment may comprise: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:17 with one, two or three amino acid additions, deletions or substitutions). , or may consist essentially of, or may consist of, a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17, or SEQ ID NO: 18 (or SEQ ID NO: 18 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 19 (or SEQ ID NO: 19 with 1, 2 or 3 amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with 1, 2 or 3 amino acid additions, deletions or substitutions), and SEQ ID NO: 27 (or SEQ ID NO: 27 with 1, 2 or 3 amino acid additions, deletions or substitutions). The antigen-binding fragment may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8. The antigen-binding fragment may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16. The antigen-binding fragment may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antigen-binding fragment may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. The first antigen-binding domain may comprise an scFv capable of binding to a CD94 / NKG2A polypeptide. The first antigen-binding domain may be an IgG having the ability to bind to a CD94 / NKG2A polypeptide. The linker may comprise a linker according to FIG. 10 or FIG. 13. The second antigen-binding domain may bind to an antigen of interest. The antigen of interest may be expressed on the surface of a cancer cell or a virus-infected cell. The antigen of interest may be an antigen of interest selected from the group consisting of an EGFR polypeptide, a HER2 polypeptide, a CEACAM5 polypeptide, a CEACAM7 polypeptide, a CD19 polypeptide, a CD22 polypeptide, a CD274 polypeptide, a CD276 polypeptide, a PSMA polypeptide, a PSCA polypeptide, an ADAM10 polypeptide, a methotelin polypeptide, a GPC2 polypeptide, a FGFR polypeptide, a VEGFR polypeptide, an IGFR polypeptide, an HIV gp120 polypeptide, an HIV gp160 polypeptide, and a SARS-CoV-2 RBD polypeptide. The second antigen-binding domain may be an antigen-binding domain according to FIG. 20. The second antigen-binding domain may comprise SEQ ID NOs: 263 and 264. The second antigen-binding domain may comprise SEQ ID NOs: 265 and 266.The cell engager may comprise a third antigen-binding domain. The third antigen-binding domain may bind to a polypeptide expressed on the surface of a NK cell. The polypeptide expressed on the surface of a NK cell may be a CD16a, NKG2A, NKG2D, NKp30, NKp44 or NKp46 polypeptide. The third antigen-binding domain is an antigen-binding domain depicted in FIG. 19. The third antigen-binding domain is an antigen-binding domain depicted in FIG. 18.

[0022] In another aspect, this document features a nucleic acid comprising (or consisting essentially of, or consisting of) a nucleic acid sequence encoding at least a portion of an antibody or antigen-binding fragment, the antibody comprising: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); The antibody may comprise (or consist essentially of, or consist of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8. The antibody may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16. The antibody may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antibody may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.The antigen-binding fragment may comprise: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:17 with one, two or three amino acid additions, deletions or substitutions). , or may consist essentially of, or may consist of, a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17, or SEQ ID NO: 18 (or SEQ ID NO: 18 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 19 (or SEQ ID NO: 19 with 1, 2 or 3 amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with 1, 2 or 3 amino acid additions, deletions or substitutions), and SEQ ID NO: 27 (or SEQ ID NO: 27 with 1, 2 or 3 amino acid additions, deletions or substitutions). The antigen-binding fragment may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antigen-binding fragment may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16.The antigen-binding fragment may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 24. The antigen-binding fragment may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. The nucleic acid sequence may encode any one of the heavy chain variable domains or regions of (i) to (ii). The nucleic acid sequence may encode any one of the light chain variable domains or regions of (i) to (ii). The nucleic acid may be a viral vector. The nucleic acid may be a phagemid.

[0023] In another aspect, this document features a nucleic acid (e.g., an isolated nucleic acid) comprising (or consisting essentially of, or consisting of) a nucleic acid sequence encoding a chimeric antigen receptor as described above or a cell engager as described above. The nucleic acid can be a viral vector. The nucleic acid can be a phagemid.

[0024] In another aspect, this document features a host cell that includes the nucleic acid of either of the two preceding paragraphs.

[0025] In another aspect, this document features a host cell expressing the chimeric antigen receptor described above or the cell engager described above. The host cell can be a T cell, a stem cell, or a NK cell.

[0026] In another aspect, this document features an antibody drug conjugate (ADC) comprising (or consisting essentially of, or consisting of) an antigen-binding domain covalently linked to a drug, where the antigen-binding domain comprises an antibody or antigen-binding fragment. The antibody comprises: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions). The antibody may comprise (or may consist essentially of, or may consist of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region of (i).The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8. The antibody may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16. The antibody may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antibody may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.The antigen-binding fragment may comprise: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:17 with one, two or three amino acid additions, deletions or substitutions). , or may consist essentially of, or may consist of, a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17, or SEQ ID NO: 18 (or SEQ ID NO: 18 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 19 (or SEQ ID NO: 19 with 1, 2 or 3 amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with 1, 2 or 3 amino acid additions, deletions or substitutions), and SEQ ID NO: 27 (or SEQ ID NO: 27 with 1, 2 or 3 amino acid additions, deletions or substitutions). The antigen-binding fragment may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antigen-binding fragment may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16.The antigen-binding fragment may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antigen-binding fragment may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. The antigen-binding domain may comprise an scFv having the ability to bind to a CD94 / NKG2A polypeptide. The antigen-binding domain may be an IgG having the ability to bind to a CD94 / NKG2A polypeptide. The drug may be selected from the group consisting of BMS1166, BMS202, IL-2, and IL-12.

[0027] In another aspect, this document features a composition comprising (or consisting essentially of, or consisting of) an antibody or antigen-binding fragment, the antibody comprising: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); The antibody may comprise (or may consist essentially of, or may consist of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antibody may comprise a light chain variable domain or region of (i).The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16. The antibody may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 24. The antibody may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.The antigen-binding fragment may comprise: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:17 with one, two or three amino acid additions, deletions or substitutions). , or may consist essentially of, or may consist of, a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17, or SEQ ID NO: 18 (or SEQ ID NO: 18 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 19 (or SEQ ID NO: 19 with 1, 2 or 3 amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with 1, 2 or 3 amino acid additions, deletions or substitutions), and SEQ ID NO: 27 (or SEQ ID NO: 27 with 1, 2 or 3 amino acid additions, deletions or substitutions). The antigen-binding fragment may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antigen-binding fragment may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16.The antigen-binding fragment may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antigen-binding fragment may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. The composition may comprise an antibody. The composition may comprise an antigen-binding fragment. The composition may comprise a checkpoint inhibitor. The checkpoint inhibitor may be selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189, and ipilimumab.

[0028] In another aspect, this document features a composition comprising (or consisting essentially of, or consisting of) the cell engager described above. The composition can include a checkpoint inhibitor. The checkpoint inhibitor can be selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189, and ipilimumab.

[0029] In another aspect, this document features a composition comprising (or consisting essentially of, or consisting of) the cells described above. The composition can include a checkpoint inhibitor. The checkpoint inhibitor can be selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189, and ipilimumab.

[0030] In another aspect, this document features a composition comprising (or consisting essentially of, or consisting of) the ADC described above. The composition may include a checkpoint inhibitor. The checkpoint inhibitor may be selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189, and ipilimumab.

[0031] In another aspect, this document features a method of treating a mammal having cancer or a viral infection. The method includes (or consists essentially of, or consists of) administering to the mammal a composition of any of the preceding four paragraphs. The mammal can be a human. The cancer can be selected from the group consisting of lung cancer, prostate cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, vaginal cancer, and cervical cancer. The number of cancer cells or virally infected cells in the mammal can be reduced after the administering step.

[0032] In another aspect, this document features a method of treating a mammal having cancer or a viral infection. The method includes (or consists essentially of, or consists of) (a) administering to the mammal a composition of any of the same four preceding paragraphs referenced in the preceding paragraphs, and (b) administering to the mammal a composition comprising a checkpoint inhibitor. The mammal can be a human. The cancer can be selected from the group consisting of lung cancer, prostate cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, vaginal cancer, and cervical cancer. The checkpoint inhibitor may be selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189 and ipilimumab. The number of cancer cells or virus-infected cells in the mammal may be reduced following the administering steps (a) and (b).

[0033] In another aspect, this document features a method for conjugating a binding molecule to a CD94 / NKG2A polypeptide, the method comprising (or consisting essentially of, or consisting of) contacting a CD94 / NKG2A polypeptide with an antibody or antigen-binding fragment. The antibody comprises: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions). The antibody may comprise (or may consist essentially of, or may consist of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region of (i).The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8. The antibody may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16. The antibody may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antibody may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.The antigen-binding fragment may comprise: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two or three amino acid additions, deletions or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:17 (or SEQ ID NO:17 with one, two or three amino acid additions, deletions or substitutions). , or may consist essentially of, or may consist of, a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17, or SEQ ID NO: 18 (or SEQ ID NO: 18 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 19 (or SEQ ID NO: 19 with 1, 2 or 3 amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with 1, 2 or 3 amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with 1, 2 or 3 amino acid additions, deletions or substitutions), and SEQ ID NO: 27 (or SEQ ID NO: 27 with 1, 2 or 3 amino acid additions, deletions or substitutions). The antigen-binding fragment may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise a heavy chain variable domain or region of (i). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 8. The antigen-binding fragment may comprise a light chain variable domain or region of (i). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 16.The antigen-binding fragment may comprise a heavy chain variable domain or region of (ii). The heavy chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24. The antigen-binding fragment may comprise a light chain variable domain or region of (ii). The light chain variable domain or region may comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. The contacting step may be performed in vitro. The contacting step may be performed in vivo. The contacting step may be performed in a mammal by administering the antibody or antigen-binding fragment to the mammal. The mammal may be a human.

[0034] In another aspect, this document features a method for conjugating a binding molecule to a CD94 / NKG2A polypeptide. The method includes (or consists essentially of, or consists of) contacting a CD94 / NKG2A polypeptide with a chimeric antigen receptor described above, a cell engager described above, or an ADC described above. The contacting step can be performed in vitro. The contacting step can be performed in vivo. The contacting step can be performed in a mammal by administering the chimeric antigen receptor, cell engager, or ADC to the mammal. The mammal can be a human.

[0035] In another aspect, this document features a composition comprising (or consisting essentially of) an antibody or antibody-binding fragment, the antibody or antibody-binding fragment being capable of binding to a CD94 / NKG2A polypeptide present on an NK cell or a T cell without inhibiting the cytotoxic activity of the NK cell or the T cell. The CD94 / NKG2A polypeptide can be a human CD94 / NKG2A polypeptide. The antibody or antibody-binding fragment can bind to an NK cell. The antibody or antibody-binding fragment can bind to a T cell.In some cases, the antibody or antigen-binding fragment of the composition comprises: (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); and (ii) a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two, or three amino acid additions, deletions, or substitutions); or (ii) an antibody comprising (or consisting essentially of, or consisting of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:17 (or SEQ ID NO:17 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO:18 (or SEQ ID NO:18 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO:19 (or SEQ ID NO:19 with the addition, deletion or substitution of one, two or three amino acid), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:25 (or SEQ ID NO:25 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO:26 (or SEQ ID NO:26 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO:27 (or SEQ ID NO:27 with the addition, deletion or substitution of one, two or three amino acid). The antibody may be capable of binding to SEQ ID NO:74 and SEQ ID NO:75, SEQ ID NO:256 and SEQ ID NO:257, and / or SEQ ID NO:259. The antibody may comprise a heavy chain variable domain or region as defined in (i) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:8.The antibody may comprise the light chain variable domain or region of (i) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 16. The antibody may comprise the heavy chain variable domain or region of (ii) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 24. The antibody may comprise the light chain variable domain or region of (ii) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.In some cases, the antibody or antigen-binding fragment of the composition comprises: (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two, or three amino acid additions, deletions, or substitutions); or (ii) an antigen-binding fragment comprising (or consisting essentially of, or consisting of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 (or SEQ ID NO: 17 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO: 18 (or SEQ ID NO: 18 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO: 19 (or SEQ ID NO: 19 with the addition, deletion or substitution of one, two or three amino acid), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO: 26 (or SEQ ID NO: 26 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO: 27 (or SEQ ID NO: 27 with the addition, deletion or substitution of one, two or three amino acid). The antigen-binding fragment may have the ability to bind to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise the heavy chain variable domain or region of (i) above.The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:8. The antigen-binding fragment may comprise a light chain variable domain or region as set forth in (i) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:16. The antigen-binding fragment may comprise a heavy chain variable domain or region as set forth in (ii) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:24. The antigen-binding fragment may comprise a light chain variable domain or region as set forth in (ii) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. This paragraph may be referred to as paragraph A.

[0036] In another aspect, this document features a composition comprising an antibody or antibody-binding fragment, the antibody or antibody-binding fragment having the ability to bind to a CD94 / NKG2A polypeptide present on an NK cell or a T cell and to reduce the expression of the CD94 / NKG2A polypeptide on the surface of the NK cell or the T cell. The CD94 / NKG2A polypeptide can be a human CD94 / NKG2A polypeptide. The antibody or antibody-binding fragment can bind to an NK cell. The antibody or antibody-binding fragment can bind to a T cell.In some cases, the antibody or antigen-binding fragment of the composition comprises either (i) or (ii) the following: (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two, or three amino acid additions, deletions, or substitutions). or (ii) an antibody comprising (or consisting essentially of, or consisting of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:17 (or SEQ ID NO:17 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO:18 (or SEQ ID NO:18 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO:19 (or SEQ ID NO:19 with the addition, deletion or substitution of one, two or three amino acid), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:25 (or SEQ ID NO:25 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO:26 (or SEQ ID NO:26 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO:27 (or SEQ ID NO:27 with the addition, deletion or substitution of one, two or three amino acid). The antibody may be capable of binding to SEQ ID NO:74 and SEQ ID NO:75, SEQ ID NO:256 and SEQ ID NO:257, and / or SEQ ID NO:259. The antibody may comprise a heavy chain variable domain or region as defined in (i) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:8.The antibody may comprise the light chain variable domain or region of (i) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 16. The antibody may comprise the heavy chain variable domain or region of (ii) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 24. The antibody may comprise the light chain variable domain or region of (ii) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.In some cases, the antibody or antigen-binding fragment of the composition comprises: (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two, or three amino acid additions, deletions, or substitutions); or (ii) an antigen-binding fragment comprising (or consisting essentially of, or consisting of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 (or SEQ ID NO: 17 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO: 18 (or SEQ ID NO: 18 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO: 19 (or SEQ ID NO: 19 with the addition, deletion or substitution of one, two or three amino acid), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with the addition, deletion or substitution of one, two or three amino acid), SEQ ID NO: 26 (or SEQ ID NO: 26 with the addition, deletion or substitution of one, two or three amino acid), and SEQ ID NO: 27 (or SEQ ID NO: 27 with the addition, deletion or substitution of one, two or three amino acid). The antigen-binding fragment may have the ability to bind to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise the heavy chain variable domain or region of (i) above.The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:8. The antigen-binding fragment may comprise a light chain variable domain or region as described in (i) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:16. The antigen-binding fragment may comprise a heavy chain variable domain or region as described in (ii) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:24. The antigen-binding fragment may comprise a light chain variable domain or region as described in (ii) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. This paragraph may be referred to as paragraph B.

[0037] In another aspect, this document features a cell engager comprising (or consisting essentially of) a first antigen-binding domain, a linker, and a second antigen-binding domain, wherein the first antigen-binding domain is capable of binding to a CD94 / NKG2A polypeptide present on a NK cell or a T cell without inhibiting the cytotoxic activity of the NK cell or the T cell. The first antigen-binding domain may comprise (or consist essentially of) an scFv capable of binding to the CD94 / NKG2A polypeptide. The first antigen-binding domain may comprise (or consist essentially of) an IgG capable of binding to the CD94 / NKG2A polypeptide.In some cases, the first antigen-binding domain is selected from the group consisting of: (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); and (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two, or three amino acid additions, deletions, or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:25 (or SEQ ID NO:25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO:26 (or SEQ ID NO:26 with one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO:27 (or SEQ ID NO:27 with one, two or three amino acid additions, deletions or substitutions). The antibody may be capable of binding to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antibody may comprise a heavy chain variable domain or region as defined in (i) above.The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:8. The antibody may comprise a light chain variable domain or region as defined in (i) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:16. The antibody may comprise a heavy chain variable domain or region as defined in (ii) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:24. The antibody may comprise a light chain variable domain or region as defined in (ii) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:32. The antibody may be a monoclonal antibody. The antibody may be an scFv antibody.In some cases, the first antigen-binding domain is selected from the group consisting of: (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); and (ii) a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two, or three amino acid additions, deletions, or substitutions); or (ii) a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:17 ( or an antigen-binding fragment comprising (or consisting essentially of, or consisting of) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 with the addition, deletion or substitution of one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with the addition, deletion or substitution of one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO: 19 (or SEQ ID NO: 19 with the addition, deletion or substitution of one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with the addition, deletion or substitution of one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with the addition, deletion or substitution of one, two or three amino acid additions, deletions or substitutions), and SEQ ID NO: 27 (or SEQ ID NO: 27 with the addition, deletion or substitution of one, two or three amino acid additions, deletions or substitutions). The antigen-binding fragment may have the ability to bind to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, and / or SEQ ID NO: 259. The antigen-binding fragment may comprise the heavy chain variable domain or region of (i) above.The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:8. The antigen-binding fragment may comprise a light chain variable domain or region as described in (i) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:16. The antigen-binding fragment may comprise a heavy chain variable domain or region as described in (ii) above. The heavy chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:24. The antigen-binding fragment may comprise a light chain variable domain or region as described in (ii) above. The light chain variable domain or region may comprise an amino acid sequence having at least 90% identity to the amino acid sequence set forth in SEQ ID NO:32. The antigen-binding fragment may be monoclonal. The antigen-binding fragment may be a Fab. The linker may comprise (or essentially consist of) a linker as described in FIG. 10 or FIG. 13. The second antigen-binding domain can bind to an antigen of interest. The antigen of interest can be expressed on the surface of a cancer cell or a virus-infected cell. The antigen of interest can be an antigen of interest selected from the group consisting of an EGFR polypeptide, a HER2 polypeptide, a CEACAM5 polypeptide, a CEACAM7 polypeptide, a CD19 polypeptide, a CD22 polypeptide, a CD274 polypeptide, a CD276 polypeptide, a PSMA polypeptide, a PSCA polypeptide, an ADAM10 polypeptide, a mesothelin polypeptide, a GPC2 polypeptide, a FGFR polypeptide, a VEGFR polypeptide, an IGFR polypeptide, an HIV gp120 polypeptide, an HIV gp160 polypeptide, and a SARS-CoV-2 RBD polypeptide. The second antigen-binding domain can be an antigen-binding domain as set forth in FIG. 20, FIG. 17B, FIG. 17C, FIG. 17D, or FIG. 17E. The second antigen-binding domain can comprise (or essentially consist of) SEQ ID NOs: 263 and 264. The second antigen-binding domain may comprise (or consist essentially of) SEQ ID NOs: 265 and 266. The cell engager may comprise a third antigen-binding domain.The third antigen-binding domain may bind to a polypeptide expressed on the surface of a NK cell. The polypeptide expressed on the surface of a NK cell may be a CD16a, NKG2A, NKG2D, NKp30, NKp44, or NKp46 polypeptide. The third antigen-binding domain may be an antigen-binding domain as set forth in FIG. 19. This paragraph may be referred to as paragraph C.

[0038] In another aspect, this document features a method for binding a molecule to a CD94 / NKG2A polypeptide on the surface of an NK cell or a T cell without inhibiting cytotoxic activity of the NK cell or the T cell. The method includes (or consists essentially of) contacting an NK cell or a T cell with any of the compositions of paragraph A or any of the cell engagers of paragraph C. The method can be in vitro. The method can be in vivo. The method can be in vivo in a mammal. The mammal can be a human.

[0039] In another aspect, this document features a method for reducing expression of a CD94 / NKG2A polypeptide on the surface of an NK cell or a T cell. The method includes (or consists essentially of) contacting an NK cell or a T cell with any of the compositions of paragraph B or any of the cell engagers of paragraph C. The method can be in vitro. The method can be in vivo. The method can be in vivo in a mammal. The mammal can be a human.

[0040] In another aspect, this document provides a method for treating HER2 +The present invention features a method for killing cancer cells. The method comprises (or consists essentially of) contacting immune cells positive for a CD94 / NKG2A polypeptide with (a) any of the compositions of paragraph A, any of the compositions of paragraph B, or any of the cell engagers of paragraph C, and (b) an anti-HER2 antibody. The method can be in vitro. The method can be in vivo. The method can be in vivo in a mammal. The mammal can be a human. The anti-HER2 antibody can be pertuzumab.

[0041] In another aspect, this document features a method of treating a mammal having cancer. The method includes (or consists essentially of) administering to the mammal any of the compositions of paragraph A, any of the compositions of paragraph B, or any of the cell engagers of paragraph C. The mammal can be a human. The cancer is HER2 + The cancer may be. The method may further comprise administering an anti-HER2 antibody to the mammal. The anti-HER2 antibody may be pertuzumab.

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Methods and materials are described herein for use in this disclosure; other suitable methods and materials known in the art may also be used. Materials, methods and examples are merely illustrative and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0043] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0044] [Brief description of the drawings]

[0045] [Figure 1] FIG. 1 shows amino acid residues 1-179 of human CD94 (SEQ ID NO:74) and 1-233 of human NKG2A polypeptide (SEQ ID NO:75). The underlined and bolded amino acid sequence of human CD94 (residues 57-179) (SEQ ID NO:256) and the underlined and bolded amino acid sequence of human NKG2A polypeptide (residues 113-233) (SEQ ID NO:257) represent the extracellular domain. FIG. 1 also shows amino acid residues 1-231 of human NKG2C polypeptide (SEQ ID NO:260). The underlined and bolded amino acid sequence of human NKG2C polypeptide (SEQ ID NO:261) represents the extracellular domain. To identify CD94 / NKG2A binders, SEQ ID NO:256 was linked to SEQ ID NO:257 using a linker between the CD94 and NKG2A sequences (SEQ ID NO:258) to create SEQ ID NO:259. To identify CD94 / NKG2A binders, SEQ ID NO:256 was linked to SEQ ID NO:261 using a linker between the CD94 and NKG2C sequences (SEQ ID NO:258) to create SEQ ID NO:262. [Figure 2A] FIG. 2A shows the amino acid sequence of the heavy chain variable domain (FIG. 2A) and the light chain variable domain (FIG. 2B) of the Fab designated clone #1 (1B2). The respective CDRs, framework sequences, and constant domains are also provided and depicted. [Figure 2B] FIG. 2A shows the amino acid sequence of the heavy chain variable domain (FIG. 2A) and the light chain variable domain (FIG. 2B) of the Fab designated clone #1 (1B2). The respective CDRs, framework sequences, and constant domains are also provided and depicted. [Figure 3A] 3A and 3B show the amino acid sequences of the heavy chain variable domain (FIG. 3A) and the light chain variable domain (FIG. 3B) of the Fab designated clone #2 (1B2-6). The respective CDRs, framework sequences, and constant domains are also provided and depicted. [Figure 3B]3A and 3B show the amino acid sequences of the heavy chain variable domain (FIG. 3A) and the light chain variable domain (FIG. 3B) of the Fab designated clone #2 (1B2-6). The respective CDRs, framework sequences, and constant domains are also provided and depicted. [Figure 4] FIG. 1 shows the nucleic acid sequences encoding the indicated strands / domains of clones #1-#2. [Figure 5-1] FIG. 1 depicts the structure of an exemplary Ig molecule and provides exemplary hinge, CH2, and CH3 regions / domains and complete Ig amino acid and nucleic acid sequences. [Figure 5-2] FIG. 1 depicts the structure of an exemplary Ig molecule and provides exemplary hinge, CH2, and CH3 regions / domains and complete Ig amino acid and nucleic acid sequences. [Figure 5-3] FIG. 1 depicts the structure of an exemplary Ig molecule and provides exemplary hinge, CH2, and CH3 regions / domains and complete Ig amino acid and nucleic acid sequences. [Figure 5-4] FIG. 1 depicts the structure of an exemplary Ig molecule and provides exemplary hinge, CH2, and CH3 regions / domains and complete Ig amino acid and nucleic acid sequences. [Figure 6A] Figure 6A shows an exemplary scFv structure. Figures 6B and 6C show the amino acid sequences of an exemplary heavy chain variable domain (Figure 6B) and an exemplary light chain variable domain (Figure 6C) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in Figure 10. Figures 6D-6G show exemplary scFv structures and provide amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 6B]Figure 6A shows an exemplary scFv structure. Figures 6B and 6C show the amino acid sequences of an exemplary heavy chain variable domain (Figure 6B) and an exemplary light chain variable domain (Figure 6C) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in Figure 10. Figures 6D-6G show exemplary scFv structures and provide amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 6C] Figure 6A shows an exemplary scFv structure. Figures 6B and 6C show the amino acid sequences of an exemplary heavy chain variable domain (Figure 6B) and an exemplary light chain variable domain (Figure 6C) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in Figure 10. Figures 6D-6G show exemplary scFv structures and provide amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 6D] Figure 6A shows an exemplary scFv structure. Figures 6B and 6C show the amino acid sequences of an exemplary heavy chain variable domain (Figure 6B) and an exemplary light chain variable domain (Figure 6C) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in Figure 10. Figures 6D-6G show exemplary scFv structures and provide amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 6E]Figure 6A shows an exemplary scFv structure. Figures 6B and 6C show the amino acid sequences of an exemplary heavy chain variable domain (Figure 6B) and an exemplary light chain variable domain (Figure 6C) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in Figure 10. Figures 6D-6G show exemplary scFv structures and provide amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 6FG] Figure 6A shows an exemplary scFv structure. Figures 6B and 6C show the amino acid sequences of an exemplary heavy chain variable domain (Figure 6B) and an exemplary light chain variable domain (Figure 6C) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in Figure 10. Figures 6D-6G show exemplary scFv structures and provide amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 7A] 7A-7B show amino acid sequences of an exemplary heavy chain variable domain (FIG. 7A) and an exemplary light chain variable domain (FIG. 7B) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in FIG. 10. FIG. 7C shows the structure of an exemplary scFv, providing amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 7B]7A-7B show amino acid sequences of an exemplary heavy chain variable domain (FIG. 7A) and an exemplary light chain variable domain (FIG. 7B) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in FIG. 10. FIG. 7C shows the structure of an exemplary scFv, providing amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 7C] 7A-7B show amino acid sequences of an exemplary heavy chain variable domain (FIG. 7A) and an exemplary light chain variable domain (FIG. 7B) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in FIG. 10. FIG. 7C shows the structure of an exemplary scFv, providing amino acid sequences with the linker, CDR, and framework sequences depicted. [Figure 8A] FIG. 8A shows the amino acid sequences of an exemplary heavy chain variable domain (FIG. 8A) and an exemplary light chain variable domain (FIG. 8B) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in FIG. 10. [Figure 8B] FIG. 8A shows the amino acid sequences of an exemplary heavy chain variable domain (FIG. 8A) and an exemplary light chain variable domain (FIG. 8B) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in FIG. 10. [Figure 9A]9A-9B show the amino acid sequences of an exemplary heavy chain variable domain (FIG. 9A) and an exemplary light chain variable domain (FIG. 9B) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in FIG. 10. [Figure 9B] 9A-9B show the amino acid sequences of an exemplary heavy chain variable domain (FIG. 9A) and an exemplary light chain variable domain (FIG. 9B) of an exemplary scFv. The respective CDR and framework sequences are also depicted. The heavy and light chain variable domains can be linked together to form an scFv using exemplary linker amino acid sequences, such as those set forth in FIG. 10. [Figure 10] 1 shows exemplary linker amino acid sequences that can be used to link heavy and light chain variable domains together to form scFvs. These linker sequences can also be used to create CARs and cell engagers. [Figure 11A] Figure 11A shows the structure of an exemplary CAR. Figure 11B is a schematic diagram of an exemplary CAR construct designed to express a CAR. A promoter sequence (e.g., a CMV immediate early promoter sequence) may be followed by a signal peptide sequence (e.g., a GM-CSF signal peptide sequence), followed by an scFv provided herein (e.g., an scFv designed to include two sets of three CDRs of an antigen-binding fragment provided herein, e.g., CDR1, CDR2, and CDR3 of the heavy chain, and CDR1, CDR2, and CDR3 of the light chain (in either order), e.g., SEQ ID NOs: 1-3 and 9-11, or SEQ ID NOs: 17-19 and 25-27), followed by an optional linker (not shown), followed by an optional hinge (e.g., a CD8 hinge sequence, not shown), followed by a transmembrane sequence (e.g., a CD8 transmembrane sequence), followed by one or more intracellular signaling domain sequences (e.g., a 4-1BB (CD137) intracellular signaling domain sequence and a CD3 zeta intracellular signaling domain sequence). [Figure 11B]Figure 11A shows the structure of an exemplary CAR. Figure 11B is a schematic diagram of an exemplary CAR construct designed to express a CAR. A promoter sequence (e.g., a CMV immediate early promoter sequence) may be followed by a signal peptide sequence (e.g., a GM-CSF signal peptide sequence), followed by an scFv provided herein (e.g., an scFv designed to include two sets of three CDRs of an antigen-binding fragment provided herein, e.g., CDR1, CDR2, and CDR3 of the heavy chain, and CDR1, CDR2, and CDR3 of the light chain (in either order), e.g., SEQ ID NOs: 1-3 and 9-11, or SEQ ID NOs: 17-19 and 25-27), followed by an optional linker (not shown), followed by an optional hinge (e.g., a CD8 hinge sequence, not shown), followed by a transmembrane sequence (e.g., a CD8 transmembrane sequence), followed by one or more intracellular signaling domain sequences (e.g., a 4-1BB (CD137) intracellular signaling domain sequence and a CD3 zeta intracellular signaling domain sequence). [Figure 12] FIG. 1 shows the amino acid sequences of exemplary signal peptides that can be used to design CARs. [Figure 13] FIG. 1 shows the amino acid sequences of exemplary hinges that can be used to design CARs. [Figure 14] FIG. 1 shows the amino acid sequences of exemplary transmembrane domains that can be used to design CARs. [Figure 15] FIG. 1 shows the amino acid sequences of exemplary intracellular signaling domains that can be used to design CARs. [Figure 16A] Figure 16A shows the amino acid sequence of a CAR (CAR #1) designed to include an scFv created using the CDRs of the Fab of clone #1. The various components of this CAR (e.g., domains and linkers) are provided and depicted. Figure 16B shows the amino acid sequence of a CAR (CAR #2) designed to include an scFv created using the CDRs of the Fab of clone #2. The various components of this CAR (e.g., domains and linkers) are provided and depicted. [Figure 16B]Figure 16A shows the amino acid sequence of a CAR (CAR #1) designed to include an scFv created using the CDRs of the Fab of clone #1. The various components of this CAR (e.g., domains and linkers) are provided and depicted. Figure 16B shows the amino acid sequence of a CAR (CAR #2) designed to include an scFv created using the CDRs of the Fab of clone #2. The various components of this CAR (e.g., domains and linkers) are provided and depicted. [Figure 17A] FIG. 17A is a schematic diagram of an exemplary BiTE or BiKE designed using the CDR1, CDR2, and CDR3 of the heavy chain provided herein in an Ig format (e.g., IgG1 format) and the CDR1, CDR2, and CDR3 of the light chain provided herein. The anti-CEACAM5 scFv (or any binding agent that targets an antigen of interest) can be linked to the C-terminus of the light chain via an optional linker (e.g., a (G4S)3 linker). FIG. 17B shows the amino acid sequence of the anti-CEACAM5 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17B also shows the nucleic acid sequence encoding the scFv. FIG. 17C shows the amino acid sequence of the anti-HER2 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17C also shows the nucleic acid sequence encoding the scFv. Figure 17D shows the amino acid sequence of an anti-EGFR scFv sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17D also shows the nucleic acid sequence encoding that scFv. Figure 17E shows the amino acid sequence of an anti-HIV binding agent sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17E also shows the nucleic acid sequence encoding that binding agent. [Figure 17B]FIG. 17A is a schematic diagram of an exemplary BiTE or BiKE designed using the CDR1, CDR2, and CDR3 of the heavy chain provided herein in an Ig format (e.g., IgG1 format) and the CDR1, CDR2, and CDR3 of the light chain provided herein. The anti-CEACAM5 scFv (or any binding agent that targets an antigen of interest) can be linked to the C-terminus of the light chain via an optional linker (e.g., a (G4S)3 linker). FIG. 17B shows the amino acid sequence of the anti-CEACAM5 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17B also shows the nucleic acid sequence encoding the scFv. FIG. 17C shows the amino acid sequence of the anti-HER2 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17C also shows the nucleic acid sequence encoding the scFv. Figure 17D shows the amino acid sequence of an anti-EGFR scFv sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17D also shows the nucleic acid sequence encoding that scFv. Figure 17E shows the amino acid sequence of an anti-HIV binding agent sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17E also shows the nucleic acid sequence encoding that binding agent. [Figure 17C]FIG. 17A is a schematic diagram of an exemplary BiTE or BiKE designed using the CDR1, CDR2, and CDR3 of the heavy chain provided herein in an Ig format (e.g., IgG1 format) and the CDR1, CDR2, and CDR3 of the light chain provided herein. The anti-CEACAM5 scFv (or any binding agent that targets an antigen of interest) can be linked to the C-terminus of the light chain via an optional linker (e.g., a (G4S)3 linker). FIG. 17B shows the amino acid sequence of the anti-CEACAM5 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17B also shows the nucleic acid sequence encoding the scFv. FIG. 17C shows the amino acid sequence of the anti-HER2 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17C also shows the nucleic acid sequence encoding the scFv. Figure 17D shows the amino acid sequence of an anti-EGFR scFv sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17D also shows the nucleic acid sequence encoding that scFv. Figure 17E shows the amino acid sequence of an anti-HIV binding agent sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17E also shows the nucleic acid sequence encoding that binding agent. [Figure 17D]FIG. 17A is a schematic diagram of an exemplary BiTE or BiKE designed using the CDR1, CDR2, and CDR3 of the heavy chain provided herein in an Ig format (e.g., IgG1 format) and the CDR1, CDR2, and CDR3 of the light chain provided herein. The anti-CEACAM5 scFv (or any binding agent that targets an antigen of interest) can be linked to the C-terminus of the light chain via an optional linker (e.g., a (G4S)3 linker). FIG. 17B shows the amino acid sequence of the anti-CEACAM5 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17B also shows the nucleic acid sequence encoding the scFv. FIG. 17C shows the amino acid sequence of the anti-HER2 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17C also shows the nucleic acid sequence encoding the scFv. Figure 17D shows the amino acid sequence of an anti-EGFR scFv sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17D also shows the nucleic acid sequence encoding that scFv. Figure 17E shows the amino acid sequence of an anti-HIV binding agent sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17E also shows the nucleic acid sequence encoding that binding agent. [Figure 17E]FIG. 17A is a schematic diagram of an exemplary BiTE or BiKE designed using the CDR1, CDR2, and CDR3 of the heavy chain provided herein in an Ig format (e.g., IgG1 format) and the CDR1, CDR2, and CDR3 of the light chain provided herein. The anti-CEACAM5 scFv (or any binding agent that targets an antigen of interest) can be linked to the C-terminus of the light chain via an optional linker (e.g., a (G4S)3 linker). FIG. 17B shows the amino acid sequence of the anti-CEACAM5 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17B also shows the nucleic acid sequence encoding the scFv. FIG. 17C shows the amino acid sequence of the anti-HER2 scFv sequence, which can be linked to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in FIG. 17A. FIG. 17C also shows the nucleic acid sequence encoding the scFv. Figure 17D shows the amino acid sequence of an anti-EGFR scFv sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17D also shows the nucleic acid sequence encoding that scFv. Figure 17E shows the amino acid sequence of an anti-HIV binding agent sequence, which can be attached to the light chain via an optional linker (e.g., a (G4S)3 linker) as shown in Figure 17A. Figure 17E also shows the nucleic acid sequence encoding that binding agent. [Figure 18] FIG. 1 shows the amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., TriTEs) that bind to T cells. [Figure 19-1] FIG. 1 shows the amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., TriKE) that bind to NK cells. [Figure 19-2] FIG. 1 shows the amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., TriKE) that bind to NK cells. [Figure 20-1]FIG. 1 shows exemplary amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., BiTEs, BiKEs, TriTEs, and / or TriKEs) that bind to the indicated target antigens of interest. [Figure 20-2] FIG. 1 shows exemplary amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., BiTEs, BiKEs, TriTEs, and / or TriKEs) that bind to the indicated target antigens of interest. [Figure 20-3] FIG. 1 shows exemplary amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., BiTEs, BiKEs, TriTEs, and / or TriKEs) that bind to the indicated target antigens of interest. [Figure 20-4] FIG. 1 shows exemplary amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., BiTEs, BiKEs, TriTEs, and / or TriKEs) that bind to the indicated target antigens of interest. [Figure 20-5] FIG. 1 shows exemplary amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., BiTEs, BiKEs, TriTEs, and / or TriKEs) that bind to the indicated target antigens of interest. [Figure 20-6] FIG. 1 shows exemplary amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., BiTEs, BiKEs, TriTEs, and / or TriKEs) that bind to the indicated target antigens of interest. [Figure 20-7] FIG. 1 shows exemplary amino acid sequences of exemplary antigen-binding domains that can be used to design cell engagers (e.g., BiTEs, BiKEs, TriTEs, and / or TriKEs) that bind to the indicated target antigens of interest. [Figure 21]FIG. 1 shows the amino acid sequence of an exemplary BiTE and / or BiKE cell engager that comprises the CDRs of clone #1 or clone #2 and targets an EGFR polypeptide in addition to binding to a CD94 / NKG2A polypeptide. [Figure 22] Schematic diagrams of different formats of exemplary cell engagers (e.g., BiTEs or BiKEs) designed to contain CDR1, CDR2, and CDR3 of a heavy chain provided herein, and / or CDR1, CDR2, and CDR3 of a light chain provided herein. [Figure 23] FIG. 1 includes graphs plotting the binding of 1B2 (Fab clone #1) and 2A8 (negative control Fab) to recombinant CD94 / NKG2A or CD94 / NKG2C polypeptides. [Figure 24-1] Figure 1 includes flow cytometry analysis of 1B2 (Fab clone #1) and 2A8 (negative control Fab) binding to primary NK cells. AAG refers to human IgG1 Fc with L234A, L235A, and P329G mutations. [Figure 24-2] Figure 1 includes flow cytometry analysis of 1B2 (Fab clone #1) and 2A8 (negative control Fab) binding to primary NK cells. AAG refers to human IgG1 Fc with L234A, L235A, and P329G mutations. [Figure 24-3] Figure 1 includes flow cytometry analysis of 1B2 (Fab clone #1) and 2A8 (negative control Fab) binding to primary NK cells. AAG refers to human IgG1 Fc with L234A, L235A, and P329G mutations. [Diagram 25] Figure 1 includes a graph plotting the equilibrium dissociation constants (KD) from a BLItz Biolayer Interferometry (BLI) system for 1B2 (Fab clone #1) and a monalizumab analog (Mona analog). LALA-PG also refers to human IgG1 Fc with L234A, L235A, and P329G mutations. [Figure 26]FIG. 1 includes graphs plotting competitive ELISA binding assessment of (1) IgG of clone #1, (2) monalizumab analogs, and (3) isotype controls as competitors for binding of HLA-E tetramer to CD94 / NKG2A or CD94 / NKG2C polypeptides. [Figure 27] FIG. 1 includes graphs plotting the binding of 1B2 (IgG clone #1) and 1B2-6 (IgG clone #2) to CD94 / NKG2A or CD94 / NKG2C polypeptides. [Figure 28] FIG. 1 includes graphs plotting competitive ELISA binding assessment of (1) IgG of clone #1, (2) IgG of clone #2, (3) monalizumab analogs, and (4) isotype control as competitors for HLA-E tetramer binding to CD94 / NKG2A polypeptide. [Figure 29] Figure 1 includes flow cytometry binding analysis of IgG clone #1, IgG clone #2, or monalizumab analog (Mona analog) to primary NK cells. Control is secondary antibody only (Secondary only). [Diagram 30] FIG. 1 includes graphs plotting binding of IgG clone #1, IgG clone #2, or Monalizumab Analog (Mona Analog) antibody (10 nM) to NKG2A-positive primary NK cells and NKG2A-negative Farage cells. Control is secondary antibody only (Secondary Only). [Diagram 31] FIG. 1 includes graphs plotting NKG2A expression levels on IL-2-activated NK cells alone or in association with HLA-E+ cancer cells following 24 hour treatment with the indicated antibodies (100 nM). [Diagram 32] FIG. 1 includes graphs plotting IFNγ and granzyme B (GrzB) secretion from IL-2-activated NKG2A+ NK cells alone or with HLA-E+ cancer cells after 24 hours of treatment with the indicated antibodies (100 nM), using intracellular staining. [Diagram 33]FIG. 1 includes graphs plotting binding of CD94 / NKG2A polypeptides and bispecific NK cell engagers (BINKs) designed to bind to the CEACAM5 target antigen of interest. Binding and specificity were tested using recombinant polypeptides: (1) CD94 / NKG2A polypeptide, (2) CD94 / NKG2C polypeptide, (3) CEACAM5-A3B3 domain, and (4) CEACAM6-AB domain. [Diagram 34] FIG. 34A is a schematic diagram of an exemplary bispecific NK cell engager (BiNK) (referred to as "BiNK (anti-HER2×anti-NKG2A)") designed to simultaneously bind a CD94 / NKG2A polypeptide and a HER2 target antigen of interest. As shown, BiNK (anti-HER2×anti-NKG2A) simultaneously binds to (a) a biotinylated CD94 / NKG2A-Fc fusion polypeptide and (b) a HER2 polypeptide immobilized on a streptavidin-coated sensor. FIG. 34B is a graph plotting the results of a BLItz assay showing loading of a biotinylated CD94 / NKG2A-Fc fusion polypeptide followed by binding of BiNK (anti-HER2×anti-NKG2A) followed by binding of a HER2 polypeptide. [Diagram 35]FIG. 35A is a graph plotting the binding of (a) an exemplary bispecific NK cell engager (BiNK) designed to simultaneously bind a CD94 / NKG2A polypeptide and a HER2 target antigen of interest (termed "BiNK (anti-HER2 x anti-NKG2A)", 100 nM), (b) an exemplary bispecific NK cell engager (BiNK) designed to simultaneously bind a CD94 / NKG2A polypeptide and an EGFR target antigen of interest (termed "BiNK (anti-EGFR x anti-NKG2A)", 100 nM), and (c) anti-CD94 / NKG2A clone #2 as an IgG molecule to primary NK cells that are NKG2A+ (termed "clone #2 IgG (anti-NKG2A)", 100 nM) compared to the binding of an hIgG1 isotype control (100 nM). BiNK (anti-HER2 x anti-NKG2A), BiNK (anti-EGFR x anti-NKG2A), and clone #2 IgG (anti-NKG2A) each showed binding to primary NK cells that exceeded that observed for the isotype control. Figure 35B is a graph plotting the binding of (a) 100 nM BiNK (anti-HER2 x anti-NKG2A), (b) 100 nM BiNK (anti-EGFR x anti-NKG2A), and (c) 100 nM cetuximab (a chimeric monoclonal antibody that binds EGFR) to A549 cells (which are non-small cell lung cancer (NSCLC) cells that are EGFR+ and HER2+) or Farage cells (which are EGFR-, HER2-, and NKG2A-) compared to the binding of hIgG1 isotype control (100 nM). BiNK (anti-HER2 x anti-NKG2A), BiNK (anti-EGFR x anti-NKG2A), and cetuximab each showed binding to A549 cells that exceeded that observed for the isotype control, while BiNK (anti-HER2 x anti-NKG2A), BiNK (anti-EGFR x anti-NKG2A), and cetuximab showed binding to Farage cells similar to that observed for the isotype control. The x-axis of the graph is Alexa647 intensity and the y-axis of the graph is cell number. [Diagram 36]FIG. 1 includes graphs plotting the percent cytotoxicity observed using an LDH release assay for primary NK cells (effector cells) incubated with A549 or 293T target cells (E:T ratio=5:1) in the presence of the indicated amounts of (a) BiNK (anti-EGFR×anti-NKG2A), (b) BiNK (anti-HER2×anti-NKG2A), (c) cetuximab, (d) pertuzumab (antibody that binds HER2), or (e) hIgG1 isotype control. The use of BiNK (anti-EGFR×anti-NKG2A), BiNK (anti-HER2×anti-NKG2A), cetuximab, and pertuzumab resulted in increased killing of A549 target cells by NK cells compared to the killing observed with the isotype control, while the use of cetuximab resulted in increased killing of 293T target cells by NK cells compared to the killing observed with the isotype control. [Figure 37] FIG. 37 includes graphs plotting CD16A-positive, NKG2A-positive, and NKG2C-positive populations (percentages after normalization to vehicle treatment) of CD56-positive NK cells in 24-hour co-cultures with A549 cells following treatment with 50 IU / mL IL-2 and 100 nM (a) BiNK (anti-EGFR x anti-NKG2A), referred to as "BiNK" in the graph in FIG. 37, (b) Clone #2 IgG (anti-NKG2A), referred to as "Clone #2" in the graph in FIG. 37, and (c) Cetuximab. Each symbol represents values ​​obtained from individual healthy donors. * p<0.05; ** p<0.01; and *** p<0.001. When NK cells were treated with BiNK (anti-EGFR x anti-NKG2A) or clone #2 IgG (anti-NKG2A), the percentage of NKG2A positive cells that were CD56 positive NK cells was reduced compared to vehicle. [Figure 38]Figure 38A is a schematic diagram of a treatment schedule for performing a cell killing assay using two doses. Figure 38B is a graph plotting the percent cytotoxicity observed using an LDH release assay for primary NK cells (effector cells) incubated with A549 target cells in the presence of (a) vehicle, (b) BiNK (anti-HER2 x anti-NKG2A), (c) clone #2 IgG (anti-NKG2A), referred to as "Clone #2" in the graph in Figure 38B, or (d) Pertuzumab (Ptz) at the indicated concentrations starting on day 0, and subsequently on day 1 in the presence of vehicle or Pertuzumab (Ptz) at the indicated concentrations. [Figure 39] Co-cultured with A549 cells and treated with (a) vehicle, (b) 100 nM cetuximab, (c) bivalent, 100 nM Clone #2 IgG (anti-NKG2A), graphed as "Clone #2" in FIG. 39, (d) 100 nM BiNK (anti-EGFR x anti-NKG2A), graphed as "BiNK" in FIG. 39, (e) monovalent, 100 nM Clone #2 Fab (anti-NKG2A), graphed as "Clone #2 Fab" in FIG. 39, (f) octavalent, 100 nM Clone #2 IgG (anti-NKG2A) mixed with SA (streptavidin), graphed as "Clone #2-SA mix" in FIG. 39, or (g) multivalent, 100 nM Clone #2 IgG, graphed as "Clone #2-SA bead mix" in FIG. NKG2A expression levels (fold change in mean fluorescence intensity (MFI)) of IL-2 (50 IU / mL) activated NK cells treated for 24 hours with a mixture of (anti-NKG2A) and SA (streptavidin) beads. Each symbol represents the value obtained from an individual healthy donor. **p<0.01. [Diagram 40]IL-2 activation (50 IU / mL), co-culture with A549 cells, and (a) 100 nM cetuximab, (b) 100 nM Clone #2 IgG (anti-NKG2A), bivalent, referred to as "Clone #2" in the graph in FIG. 40, (c) 100 nM BiNK (anti-EGFR x anti-NKG2A), referred to as "BiNK" in the graph in FIG. 40, (d) 100 nM Clone #2 Fab (anti-NKG2A), monovalent, referred to as "Clone #2 Fab" in the graph in FIG. 40, (e) 100 nM Clone #2 IgG, referred to as "Clone #2-SA mix" in the graph in FIG. 40. FIG. 40 includes graphs plotting the percentage of IFNγ+, TNFα+, or Granzyme B+ (GrzB+) cells (normalized to vehicle) of NK cells 24 hours after treatment with (f) a mixture of 100 nM Clone #2 IgG (anti-NKG2A) with SA (streptavidin) beads, or (g) a mixture of 100 nM Clone #2 IgG (anti-NKG2A) with SA (streptavidin) beads, which is multivalent and referred to as "Clone #2-SA Bead Mix" in the graph of FIG. 40. Each symbol represents the value obtained from an individual healthy donor. **p<0.01; ***p<0.001; and ****p<0.0001. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0046] Detailed Description This document provides binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and ADCs) that bind (e.g., specifically bind) to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). For example, this document provides binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and ADCs) that bind (e.g., specifically bind) to a polypeptide comprising, consisting essentially of, or consisting of the amino acids set forth in SEQ ID NO:74 and SEQ ID NO:75, SEQ ID NO:256 and SEQ ID NO:257, and / or SEQ ID NO:259 (see, e.g., FIG. 1). In some cases, a binding agent provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, or ADCs) can have the ability to bind to a CD94 / NKG2A polypeptide and can lack the ability to bind to a CD94 / NKG2C heterodimeric polypeptide (e.g., SEQ ID NO:262). For example, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, or ADC) can have the ability to bind to a human CD94 / NKG2A polypeptide and can lack the ability to bind to a human CD94 / NKG2C polypeptide (e.g., SEQ ID NO: 262).

[0047] The term "antibody" as used herein includes polyclonal antibodies, monoclonal antibodies, recombinant antibodies, humanized antibodies, human antibodies, chimeric antibodies, multispecific antibodies (e.g., bispecific antibodies) formed from at least two antibodies, diabodies, single chain variable fragment antibodies (e.g., scFv antibodies), and tandem single chain variable fragment antibodies (e.g., taFv). Diabodies can include two chains each having a heavy chain variable domain and a light chain variable domain derived from the same or different antibodies (see, e.g., Hornig and Farber-Schwarz, Methods Mol. Biol., 907:713-27 (2012); and Brinkmann and Kontermann, MAbs., 9(2):182-212 (2017)). The two variable regions can be connected by a polypeptide linker (e.g., a polypeptide linker having a length of 5 to 10 residues or a polypeptide linker described in FIG. 10). In some cases, there may be interdomain disulfide bonds in one or both of the pairs of heavy and light chain variable domains of the diabody. scFvs are single chain polypeptide antibodies in which the heavy and light chain variable domains are connected directly or via a polypeptide linker (e.g., a polypeptide linker having a length of 8-18 residues or a polypeptide linker as described in FIG. 10). See also Chen et al., Adv. Drug Deliv. Rev., 65(10):1357-1369 (2013). scFvs can be designed with an orientation in which the heavy chain variable domain is followed by the light chain variable domain, or the light chain variable domain is followed by the heavy chain variable domain. In both cases, an optional linker can be located between the two domains. Examples of scFv structures of scFvs provided herein include, but are not limited to, the structures described in FIGS. 6A-6G, 7A-7C, 8A-8B, and 9A-9B.

[0048] The antibodies provided herein may comprise the CDRs described herein (e.g., described in Table 13) and may be configured to be human, humanized, or chimeric antibodies. In some cases, the antibodies provided herein may comprise the CDRs described herein (e.g., described in Table 13) and may be monoclonal antibodies. In some cases, the antibodies provided herein may comprise the CDRs described herein (e.g., described in Table 13) and may be configured as scFv antibodies.

[0049] The term "antibody-binding fragment" as used herein refers to a fragment of an antibody (e.g., a humanized antibody fragment, a human antibody fragment, or a chimeric antibody fragment) that has the ability to bind to an antigen. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', or F(ab')2 antigen-binding fragments. The antigen-binding fragments provided herein may include CDRs described herein (e.g., as described in Table 13) and may be configured to be human antigen-binding fragments, humanized antigen-binding fragments, or chimeric antigen-binding fragments. In some cases, the antigen-binding fragments provided herein may include CDRs described herein (e.g., as described in Table 13) and may be monoclonal antigen-binding fragments. In some cases, the antigen-binding fragments provided herein may include CDRs described herein (e.g., as described in Table 13) and may be configured as a Fab antibody. In some cases, the Fab antibody may include a partial hinge sequence (e.g., SEQ ID NO: 73) for disulfide bonds between the heavy and light chains of the Fab.

[0050] The term "antibody domain" as used herein refers to a domain of an antibody, e.g., a heavy chain variable domain (VH domain) or a light chain variable domain (VL domain), in the absence of one or more other domains of the antibody. In some cases, an antibody domain may be a single antibody domain (e.g., a VH domain or a VL domain) capable of binding to an antigen. The antibody domains provided herein may comprise the CDRs described herein (e.g., as described in Table 13) and may be a human antibody domain (e.g., a human VH domain), a humanized antibody domain (e.g., a humanized VH domain), or a chimeric antibody domain (e.g., a chimeric VH domain). In some cases, the antibody domains provided herein may comprise the CDRs described herein (e.g., as described in Table 13) and may be a monoclonal antibody domain. In some cases, the antibody domains provided herein may comprise the CDRs described herein (e.g., as described in Table 13) and may be engineered as a single VH domain or a single VL domain.

[0051] The anti-CD94 / NKG2A antibodies, anti-CD94 / NKG2A antigen-binding fragments, or anti-CD94 / NKG2A antibody domains provided herein may be IgA-, IgD-, IgE-, IgG-, or IgM-type antibody domains, including IgG- or IgM-type, including but not limited to IgG1-, IgG2-, IgG3-, IgG4-, IgM1-, and IgM2-types. In some cases, the antibodies provided herein (e.g., anti-CD94 / NKG2A antibodies) may be scFv antibodies. In some cases, the antigen-binding fragments provided herein (e.g., anti-CD94 / NKG2A antibody fragments) may be Fab. In some cases, the antibodies provided herein (e.g., anti-CD94 / NKG2A antibodies) may be fully intact antibodies having the structure depicted in FIG. 5. In some cases, the antibody domains provided herein (e.g., anti-CD94 / NKG2A antibody domains) may be VH domains.

[0052] The term "chimeric antigen receptor" as used herein refers to a chimeric polypeptide designed to include an optional signal peptide, an antigen-binding domain, an optional hinge, a transmembrane domain, and one or more intracellular signaling domains. As described herein, the antigen-binding domain of the CAR provided herein can be designed to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). For example, the CAR provided herein can be designed to include components (e.g., a combination of CDRs) of an antibody, antigen-binding fragment, and / or antibody domain described herein as the antigen-binding domain, provided that the antigen-binding domain has the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). In some examples, a CAR provided herein can be designed to include an antigen-binding domain (e.g., SEQ ID NOs: 1-3 and 9-11, or SEQ ID NOs: 17-19 and 25-27) that includes two sets of three CDRs of an antigen-binding fragment provided herein (e.g., CDR1, CDR2, and CDR3 of the heavy chain, and CDR1, CDR2, and CDR3 of the light chain). In some cases, the antigen-binding domain of a CAR that targets a CD94 / NKG2A polypeptide can be designed to include a VH domain described herein or a scFv antibody described herein.

[0053] Examples of CAR structures that can be used to generate the CARs provided herein include, but are not limited to, those depicted in Figures 11A and 11B.

[0054] In some cases, the CARs provided herein may be designed to include a signal peptide. Any suitable signal peptide may be used to design the CARs described herein. Examples of signal peptides that may be used to generate the CARs described herein include, but are not limited to, signal peptides from human IGKV1-39, IGKV1-16, IGKV1-33, IGKV3-11, IGKV4-1, or IGKV6-21. In some cases, the CARs provided herein may be designed to include a signal peptide that includes, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 12. In some cases, the CARs provided herein may be designed to include a signal peptide that includes, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 12, with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof. In some cases, a CAR provided herein can be designed to include a signal peptide that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 12, with no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, or no more than 10 amino acid deletions, additions, substitutions, or combinations thereof.

[0055] In some cases, the CARs provided herein can be designed to include a hinge. Any suitable hinge can be used to design the CARs described herein. Examples of hinges that can be used to create the CARs described herein include, but are not limited to, an Ig-derived hinge (e.g., an IgG1-derived hinge, an IgG2-derived hinge, or an IgG4-derived hinge), an Ig-derived hinge containing a CD2 domain and a CD3 domain, an Ig-derived hinge containing a CD2 domain and lacking a CD3 domain, an Ig-derived hinge containing a CD3 domain and lacking a CD2 domain, an Ig-derived hinge lacking a CD2 domain and lacking a CD3 domain, a CD8α-derived hinge, a CD28-derived hinge, and a CD3ζ-derived hinge. The CARs provided herein can be designed to include any suitable length of hinge. For example, the CARs provided herein can be designed to include a hinge that is about 3 to about 75 (e.g., about 3 to about 65, about 3 to about 50, about 5 to about 75, about 10 to about 75, about 5 to about 50, about 10 to about 50, about 10 to about 40, or about 10 to about 30) amino acid residues in length. In some cases, a linker sequence may be used as a hinge to generate the CARs described herein. For example, any one of the linker sequences set forth in FIG. 10 may be used as a hinge for the CARs described herein.

[0056] In some cases, the CARs provided herein can be designed to include a hinge that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 10 or Figure 13. In some cases, the CARs provided herein can be designed to include a hinge that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 10 or Figure 13, with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof. In some cases, the CARs provided herein can be designed to include a hinge that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 10 or Figure 13, with two or less, three or less, four or less, five or less, six or less, seven or less, eight or less, nine or less, or ten or fewer amino acid deletions, additions, substitutions, or combinations thereof.

[0057] The CARs provided herein can be designed to include any suitable transmembrane domain. For example, the transmembrane domain of the CARs provided herein can be, but is not limited to, a CD3ζ transmembrane domain, a CD4 transmembrane domain, a CD8α transmembrane domain, a CD28 transmembrane domain, and a 4-1BB transmembrane domain. In some cases, the CARs provided herein can be designed to include a transmembrane domain that includes, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 14. In some cases, the CARs provided herein can be designed to include a transmembrane domain that includes, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 14, with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof. In some cases, a CAR provided herein can be designed to include a transmembrane domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 14, with no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, or no more than 10 amino acid deletions, additions, substitutions, or combinations thereof.

[0058] The CARs provided herein may be designed to include one or more intracellular signaling domains. For example, the CARs provided herein may be designed to include one, two, three, or four intracellular signaling domains. Any suitable intracellular signaling domain or combination of intracellular signaling domains may be used to generate the CARs described herein. Examples of intracellular signaling domains that can be used to generate the CARs described herein include, but are not limited to, CD3ζ intracellular signaling domain, CD27 intracellular signaling domain, CD28 intracellular signaling domain, OX40 (CD134) intracellular signaling domain, 4-1BB (CD137) intracellular signaling domain, CD278 intracellular signaling domain, DAP10 intracellular signaling domain, and DAP12 intracellular signaling domain. In some cases, the CARs described herein may be designed to be first generation CARs with CD3ζ intracellular signaling domain. In some cases, the CARs described herein may be designed to be second generation CARs with CD28 intracellular signaling domain followed by CD3ζ intracellular signaling domain. In some cases, the CARs described herein can be designed to be third generation CARs having (a) a CD28 intracellular signaling domain followed by (b) a CD27 intracellular signaling domain, an OX40 intracellular signaling domain, or a 4-1BB intracellular signaling domain followed by (c) a CD3ζ intracellular signaling domain. In some cases, the CARs provided herein can be designed to include at least one intracellular signaling domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 15.In some cases, the CARs provided herein can be designed to include at least one intracellular signaling domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 15, having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid deletions, additions, substitutions, or combinations thereof, provided that the intracellular signaling domain has at least some activity to activate intracellular signaling. In some cases, the CARs provided herein can be designed to include at least one intracellular signaling domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 15, having 2 or less, 3 or less, 4 or less, 5 or less, 6 or less, 7 or less, 8 or less, 9 or less, or 10 or less amino acid deletions, additions, substitutions, or combinations thereof, provided that the intracellular signaling domain has at least some activity to activate intracellular signaling.

[0059] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a heavy chain variable domain comprising SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, followed by a linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g. an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g. a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g. a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a heavy chain variable domain comprising SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, followed by SEQ ID NO:100, followed by a light chain variable domain comprising SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, followed by SEQ ID NO:102, followed by SEQ ID NO:113, followed by SEQ ID NO:124, followed by SEQ ID NO:129, followed by SEQ ID NO:128, followed by SEQ ID NO:97, followed by SEQ ID NO:126.

[0060] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a heavy chain variable domain comprising SEQ ID NO:8, followed by a linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO:16, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g., a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g., a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a heavy chain variable domain comprising SEQ ID NO:8, followed by SEQ ID NO:100, followed by a light chain variable domain comprising SEQ ID NO:16, followed by SEQ ID NO:102, followed by SEQ ID NO:113, followed by SEQ ID NO:124, followed by SEQ ID NO:129, followed by SEQ ID NO:128, followed by SEQ ID NO:97, followed by SEQ ID NO:126.

[0061] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a light chain variable domain comprising SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, followed by a linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g. an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g. a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g. a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a light chain variable domain comprising SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, followed by SEQ ID NO:100, followed by a heavy chain variable domain comprising SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, followed by SEQ ID NO:102, followed by SEQ ID NO:113, followed by SEQ ID NO:124, followed by SEQ ID NO:129, followed by SEQ ID NO:128, followed by SEQ ID NO:97, followed by SEQ ID NO:126 (see, e.g., Figure 16).

[0062] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a light chain variable domain comprising SEQ ID NO: 16, followed by a linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO: 8, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g. an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g. a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g. a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a light chain variable domain comprising SEQ ID NO: 16, followed by SEQ ID NO: 100, followed by a heavy chain variable domain comprising SEQ ID NO: 8, followed by SEQ ID NO: 102, followed by SEQ ID NO: 113, followed by SEQ ID NO: 124, followed by SEQ ID NO: 129, followed by SEQ ID NO: 128, followed by SEQ ID NO: 97, followed by SEQ ID NO: 126 (see, e.g., Figure 16).

[0063] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a heavy chain variable domain comprising SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, followed by a linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO: 25, SEQ ID NO: 26, and SEQ ID NO: 27, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g., a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g., a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a heavy chain variable domain comprising SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19, followed by SEQ ID NO:100, followed by a light chain variable domain comprising SEQ ID NO:25, SEQ ID NO:26, and SEQ ID NO:27, followed by SEQ ID NO:102, followed by SEQ ID NO:113, followed by SEQ ID NO:124, followed by SEQ ID NO:129, followed by SEQ ID NO:128, followed by SEQ ID NO:97, followed by SEQ ID NO:126.

[0064] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a heavy chain variable domain comprising SEQ ID NO: 24, followed by a linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO: 32, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g. an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g. a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g. a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a heavy chain variable domain comprising SEQ ID NO:24, followed by SEQ ID NO:100, followed by a light chain variable domain comprising SEQ ID NO:32, followed by SEQ ID NO:102, followed by SEQ ID NO:113, followed by SEQ ID NO:124, followed by SEQ ID NO:129, followed by SEQ ID NO:128, followed by SEQ ID NO:97, followed by SEQ ID NO:126.

[0065] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a light chain variable domain comprising SEQ ID NO:25, SEQ ID NO:26, and SEQ ID NO:27, followed by a linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g. an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g. a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g. a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a light chain variable domain comprising SEQ ID NO:25, SEQ ID NO:26, and SEQ ID NO:27, followed by SEQ ID NO:100, followed by a heavy chain variable domain comprising SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19, followed by SEQ ID NO:102, followed by SEQ ID NO:113, followed by SEQ ID NO:124, followed by SEQ ID NO:129, followed by SEQ ID NO:128, followed by SEQ ID NO:97, followed by SEQ ID NO:126 (see, e.g., Figure 16B).

[0066] In some cases, a CAR targeting a CD94 / NKG2A polypeptide can be designed to include an scFv having a light chain variable domain comprising SEQ ID NO: 32, followed by a linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO: 24, followed by a hinge, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g. an IgG4-derived hinge, a CD8α hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain, such as a transmembrane domain described in FIG. 14 (e.g. a human CD28 transmembrane domain or a CD8α transmembrane domain), followed by one or more intracellular signaling domains, such as one or more intracellular signaling domains described in FIG. 15 (e.g. a human 4-1BB intracellular signaling domain followed by a human CD3ζ intracellular signaling domain). For example, a CAR targeting a CD94 / NKG2A polypeptide can be designed to comprise an scFv having a light chain variable domain comprising SEQ ID NO:32, followed by SEQ ID NO:100, followed by a heavy chain variable domain comprising SEQ ID NO:24, followed by SEQ ID NO:102, followed by SEQ ID NO:113, followed by SEQ ID NO:124, followed by SEQ ID NO:129, followed by SEQ ID NO:128, followed by SEQ ID NO:97, followed by SEQ ID NO:126 (see, e.g., Figure 16B).

[0067] As used herein, the term "cell engager" refers to a polypeptide that includes two or more antigen-binding domains (e.g., two, three, or four antigen-binding domains) and has the ability to link two cells together. Specific examples of cell engagers include, but are not limited to, BiTE, TriTE, BiKE, and TriKE. Generally, the cell engagers provided herein are capable of binding to cells (e.g., CD94 / NKG2A + CD8 + T cells and / or CD94 / NKG2A +In some cases, the cell engagers described herein may be designed to include at least one antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) expressed on the surface of a human NK cell, as well as at least one antigen-binding domain capable of binding to a target antigen of interest. + Cells (e.g., CD94 / NKG2A + CD8 + T cells and / or CD94 / NKG2A +A cell engager may link a cell (e.g., a NK cell) to another cell (e.g., a target cell expressing a target antigen of interest) via the two or more antigen binding domains of the cell engager. Specific examples of cell engager structures of cell engagers provided herein include, but are not limited to, those depicted in FIG. 17A and / or FIG. 22. In some cases, the anti-CEACAM5 scFv depicted in FIG. 17B may be replaced with a different antigen binding domain capable of binding to a target antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell), such as those depicted in FIG. 17C or FIG. 17D. Illustrative examples of antigen-binding domains capable of binding to a target antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell) include, but are not limited to, an EGFR polypeptide (see, e.g., FIG. 17D), an IGF-1R polypeptide, a HER2 polypeptide (see, e.g., FIG. 17C), a HER3 polypeptide, an EphA4 polypeptide, a PD-1 polypeptide, a PD-L1 polypeptide, a CTLA4 polypeptide, a DLL4 polypeptide, an ADAM10 polypeptide, a C-Met polypeptide, a GPC1 polypeptide, a GPC2 polypeptide, a MUC1 polypeptide, a CEACAM1 polypeptide, a CEACAM5 polypeptide (see, e.g., FIG. 17B), a CEACAM6 polypeptide, a CEACAM7 polypeptide, an EpCAM polypeptide, a CD19 polypeptide, a CD20 polypeptide, a CD22 polypeptide, a CD47 polypeptide, a VEGFR1 polypeptide, a VEGFR2 polypeptide, a CD30 polypeptide, a CD33 polypeptide, a KIT polypeptide, a FGFR polypeptide, a PDGFR polypeptide, a gp120 polypeptide (e.g., an HIV gp120 polypeptide), gp160 polypeptide (e.g., HIV gp120 polypeptide), or RBD polypeptide (e.g., SARS-CoV-2 RBD polypeptide), include antibodies, antigen-binding fragments, and antibody domains capable of binding to a gp160 polypeptide (e.g., HIV gp120 polypeptide), ... or an RBD polypeptide (e.g., a SARS-CoV-2 RBD polypeptide).Specific examples of antigen-binding domains capable of binding to a target antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell) include, but are not limited to, those depicted in Figure 20. In some cases, a binding agent, such as the HIV binding agent depicted in Figure 17E, may be used to design a cell engager capable of binding to an HIV coat protein.

[0068] As described herein, at least one antigen-binding domain of a cell engager provided herein may be designed to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and at least another antigen-binding domain of a cell engager provided herein may be designed to bind to a target antigen of interest (e.g., a CEACAM5 polypeptide). For example, a cell engager provided herein may be designed to include components of an antibody, antigen-binding fragment, and / or antibody domain (e.g., a combination of CDRs) described herein as an antigen-binding domain, provided that the antigen-binding domain has the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). In some specific examples, the cell engagers provided herein may be designed to include an antigen-binding domain that includes two sets of three CDRs (e.g., CDR1, CDR2, and CDR3 of the heavy chain and CDR1, CDR2, and CDR3 of the light chain) of an antigen-binding fragment provided herein (e.g., SEQ ID NOs: 1-3 and 9-11, or SEQ ID NOs: 17-19 and 25-27). In some cases, the antigen-binding domain of a cell engager that targets a CD94 / NKG2A polypeptide may be designed to include a VH domain described herein or a scFv / Fab antibody described herein. In some cases, the antigen-binding domain of a CAR described herein that has the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may be designed to include a VH domain described herein or a scFv / Fab antibody described herein. + Cells (e.g., CD94 / NKG2A + CD8+ T cells and / or CD94 / NKG2A + The antigen-binding domain of a cell engager that binds to a NK cell may be used.

[0069] As described herein, a cell engager may be designed to include at least one antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and at least one other antigen-binding domain that binds to a target antigen of interest. In some cases, a cell engager may include an antigen-binding domain capable of binding to a target antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virally infected cell), a cell (e.g., a CD94 / NKG2A polypeptide), and at least one other antigen-binding domain that binds to a target antigen of interest. + CD8 + T cells and / or CD94 / NKG2A + The TriTE and / or TriKE can comprise a first antigen-binding domain capable of binding to a target antigen of interest (e.g., a CEACAM5 polypeptide) expressed on the surface of a cell (e.g., a T cell or an NK cell), and one or more other antigen-binding domains (e.g., one, two, three, or four other antigen-binding domains), each of which may bind to a different antigen expressed on the surface of a different cell type or may bind to a different antigen expressed on the surface of the same cell type. For example, TriTE and / or TriKE can comprise a first antigen-binding domain capable of binding to a target antigen of interest (e.g., a CEACAM5 polypeptide) expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell), a T cell or an NK cell (e.g., a CD94 / NKG2A polypeptide), and one or more other antigen-binding domains (e.g., one, two, three, or four other antigen-binding domains), each of which may bind to a different antigen expressed on the surface of a different cell type or may bind to a different antigen expressed on the surface of the same cell type. + CD8 + T cells and / or CD94 / NKG2A +The antibody may be designed to have a second antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) expressed on the surface of a T cell (e.g., a CD3 polypeptide) or an antigen expressed on the surface of a NK cell (e.g., a CD16 polypeptide, such as a CD16A polypeptide).

[0070] In some cases, when designing a cell engager, such as a TriTE, for linking a CD94 / NKG2A+ T cell with a target cell expressing a target antigen of interest, the cell engager may include a first antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) expressed on the surface of a T cell, a second antigen-binding domain capable of binding to a polypeptide expressed on the surface of a T cell, and a third antigen-binding domain capable of binding to a target antigen of interest expressed on the surface of a target cell (e.g., a cancer cell and / or a virus-infected cell). Examples of polypeptides expressed on the surface of a T cell that can be targeted by the antigen-binding domain of a cell engager provided herein include, but are not limited to, a CD3 polypeptide. Examples of antigen-binding domains capable of binding to a polypeptide expressed on the surface of a T cell that can be used to generate a cell engager (e.g., a TriTE) provided herein include, but are not limited to, an anti-CD3 scFv and an anti-CD3 VH domain. Further examples of amino acid sequences that can be used as antigen-binding domains capable of binding to a polypeptide expressed on the surface of a T cell (e.g., CD3) are described in U.S. Pat. No. 6,750,325 (see, e.g., the sequence listing in U.S. Pat. No. 6,750,325).

[0071] In some cases, cell engagers provided herein (e.g., TriTEs) may be designed to include an antigen binding domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 18. In some cases, cell engagers provided herein may be designed to include an antigen binding domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 18 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen binding domain has the ability to bind to a polypeptide expressed on the surface of a T cell. In some cases, cell engagers provided herein can be designed to comprise an antigen-binding domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 18, with no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, or no more than 10 amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen-binding domain has the ability to bind to a polypeptide expressed on the surface of a T cell.

[0072] In some cases, cell engagers, such as CD94 / NKG2A +When designing TriKE to link NK cells to target cells expressing a target antigen of interest, the cell engager may comprise a first antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) expressed on the surface of an NK cell, a second antigen-binding domain capable of binding to a polypeptide expressed on the surface of an NK cell, and an antigen-binding domain capable of binding to a target antigen of interest expressed on the surface of a target cell (e.g., a cancer cell and / or a virus-infected cell). Examples of polypeptides expressed on the surface of NK cells that can be targeted by the antigen-binding domains of the cell engagers provided herein include, but are not limited to, CD16 polypeptides (e.g., CD16a polypeptides), NKG2A polypeptides, NKG2D polypeptides, NKp30 polypeptides, NKp44 polypeptides, and NKp46 polypeptides. Examples of antigen-binding domains capable of binding to a polypeptide expressed on the surface of a NK cell that can be used to generate a cell engager (e.g., TriKE) provided herein include, but are not limited to, an anti-CD16a scFv, an anti-NKG2A scFv, an anti-NKG2D scFv, an anti-NKp30 scFv (see, e.g., BioLegend Catalog #325207), an anti-NKp44 scFv, an anti-NKp46 scFv, an anti-CD16a VH domain, an anti-NKG2A VH domain, an anti-NKG2D VH domain, an anti-NKp30 VH domain, an anti-NKp44 VH domain, and an anti-NKp46 VH domain.Further examples of amino acid sequences that can be used as antigen-binding domains capable of binding to a polypeptide expressed on the surface of a NK cell (e.g., CD16, NKG2A, NKG2D, or NKp46) are described in McCall et al., (Mol. Immunol., 36(7):433-445 (1999); e.g., anti-CD16 scFv sequences); International Patent Application Publication No. PCT / US2017 / 048721 (see, e.g., CDRs and sequence listing for anti-CD16a binding domains); U.S. Patent Application Publication No. 2011 / 0052606 (see, e.g., CDRs and sequence listing for anti-NKG2A antibodies, e.g., Z199); U.S. Patent Application Publication No. 2011 / 0150870 (see, e.g., CDRs and sequence listing for anti-NKG2D antibodies); U.S. Patent Application Publication No. 2018 / 0369373 (see, e.g., CDRs and sequence listing for anti-NKp46 antibodies); and U.S. Patent Application Publication No. 2017 / 0368169 (see, e.g., CDRs and sequence listing for anti-NKp46 antibodies).

[0073] In some cases, cell engagers provided herein (e.g., TriTEs) can be designed to include an antigen binding domain (e.g., scFv or VH) that comprises, consists essentially of, or consists of one or more of the amino acid sequences set forth in Figure 19. In some cases, cell engagers provided herein can be designed to include an antigen binding domain (e.g., scFv or VH) that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 19 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen binding domain has the ability to bind to a polypeptide expressed on the surface of a NK cell. In some cases, the cell engagers provided herein can be designed to comprise an antigen binding domain (e.g., scFv or VH) that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in FIG. 19, with no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, or no more than 10 amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen binding domain has the ability to bind to a polypeptide expressed on the surface of a NK cell.

[0074] In some cases, the cell engagers provided herein may be designed to include a linker located between each antigen-binding domain. Any suitable linker may be used to design the cell engagers provided herein. Examples of linkers that can be used to create the cell engagers described herein include, but are not limited to, the linker sequences described in Figure 10. The cell engagers provided herein may be designed to include a linker of any suitable length. For example, cell engagers provided herein can be designed to include a linker of about 3 to about 100 amino acid residues in length (e.g., about 3 to about 90, about 3 to about 80, about 3 to about 70, about 3 to about 60, about 3 to about 50, about 3 to about 40, about 3 to about 30, about 3 to about 20, about 3 to about 15, about 5 to about 100, about 10 to about 100, about 20 to about 100, about 30 to about 100, about 40 to about 100, about 50 to about 100, about 60 to about 100, about 70 to about 100, about 10 to about 50, about 10 to about 40, about 10 to about 30, about 10 to about 20, or about 12 to about 17). In some cases, cell engagers provided herein (e.g., BiTEs) can be designed to include a GGGGSGGGSGGGGS (SEQ ID NO: 78) linker. In some cases, the hinge of a CAR described herein can be used as a linker to generate a cell engager described herein. For example, any one of the sequences described in Figure 13 can be used as a linker of a cell engager described herein.

[0075] In some cases, cell engagers provided herein may be designed to include a linker that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 10 or Figure 13. In some cases, cell engagers provided herein may be designed to include a linker that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 10 or Figure 13, with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid deletions, additions, substitutions, or combinations thereof. In some cases, cell engagers provided herein may be designed to include a linker that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Figure 10 or Figure 13, with 2 or less, 3 or less, 4 or less, 5 or less, 6 or less, 7 or less, 8 or less, 9 or less, or 10 or less amino acid deletions, additions, substitutions, or combinations thereof.

[0076] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include a heavy chain variable domain comprising SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, followed by an optional linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO:78), followed by an scFv having an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0077] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include a light chain variable domain comprising SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, followed by an optional linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO:78), followed by an scFv having an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0078] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include a heavy chain variable domain comprising SEQ ID NO:8, followed by an optional linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO:16, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO:78), followed by an scFv having an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0079] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include an scFv having a light chain variable domain comprising SEQ ID NO: 16, followed by a linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO: 8, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO: 78), followed by an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0080] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include a heavy chain variable domain comprising SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19, followed by an optional linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO:25, SEQ ID NO:26, and SEQ ID NO:27, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO:78), followed by an scFv having an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0081] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include a light chain variable domain comprising SEQ ID NO:25, SEQ ID NO:26, and SEQ ID NO:27, followed by an optional linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO:78), followed by an scFv having an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0082] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include an scFv having a heavy chain variable domain comprising SEQ ID NO: 24, followed by an optional linker, such as a linker described in FIG. 10, followed by a light chain variable domain comprising SEQ ID NO: 32, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO: 78), followed by an antigen binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0083] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to include an scFv having a light chain variable domain comprising SEQ ID NO:32, followed by an optional linker, such as a linker described in FIG. 10, followed by a heavy chain variable domain comprising SEQ ID NO:24, followed by an optional linker, such as a hinge / linker described in FIG. 10 or FIG. 13 (e.g., SEQ ID NO:78), followed by an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0084] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to comprise an IgG (e.g., IgG1) configuration having (a) a heavy chain comprising, consisting essentially of, or consisting of a heavy chain variable domain comprising SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, an Ig hinge, and a constant domain (e.g., CH1, CH2, and CH3 domains), and (b) a light chain comprising, consisting essentially of, or consisting of a light chain variable domain comprising SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, a constant domain (e.g., a kappa or lambda constant domain), and an antigen binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0085] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to comprise an IgG (e.g., IgG1) configuration having (a) a heavy chain comprising, consisting essentially of, or consisting of a heavy chain variable domain comprising SEQ ID NO:8, an Ig hinge, and a constant domain (e.g., CH1, CH2, and CH3 domains), and (b) a light chain comprising, consisting essentially of, or consisting of a light chain variable domain comprising SEQ ID NO:16, a constant domain (e.g., a kappa or lambda constant domain), and an antigen binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0086] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to comprise an IgG (e.g., IgG1) configuration having (a) a heavy chain comprising, consisting essentially of, or consisting of a heavy chain variable domain comprising SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19, an Ig hinge, and a constant domain (e.g., CH1, CH2, and CH3 domains), and (b) a light chain comprising, consisting essentially of, or consisting of a light chain variable domain comprising SEQ ID NO:25, SEQ ID NO:26, and SEQ ID NO:27, a constant domain (e.g., a kappa or lambda constant domain), and an antigen binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virus-infected cell).

[0087] In some cases, a cell engager (e.g., BiTE, BiKE, TriTE, and / or TriKE) targeted to an antigen of interest can be designed to comprise an IgG (e.g., IgG1) configuration having (a) a heavy chain comprising, consisting essentially of, or consisting of a heavy chain variable domain comprising SEQ ID NO:24, an Ig hinge, and a constant domain (e.g., CH1, CH2, and CH3 domains), and (b) a light chain comprising, consisting essentially of, or consisting of a light chain variable domain comprising SEQ ID NO:32, a constant domain (e.g., a kappa or lambda constant domain), and an antigen-binding domain capable of binding to an antigen of interest expressed on the surface of a cell (e.g., a cancer cell and / or a virally infected cell).

[0088] In one embodiment, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise: (i) a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO: 1 (or a variant of SEQ ID NO: 1 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO: 2 (or a variant of SEQ ID NO: 2 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO: 3 (or a variant of SEQ ID NO: 3 with one or two amino acid modifications); and / or (ii) a light chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO: 9 (or a variant of SEQ ID NO: 9 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO: 10 (or a variant of SEQ ID NO: 10 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO: 11 (or a variant of SEQ ID NO: 11 with one or two amino acid modifications). Examples of such antigen-binding fragments having these CDRs and the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) include, but are not limited to, the Fab depicted in Figures 2A and 2B.

[0089] In some cases, the antibody has the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and has (a) a CDR1 having the amino acid sequence set forth in SEQ ID NO: 1 (or a variant of SEQ ID NO: 1 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO: 2 (or a variant of SEQ ID NO: 2 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO: 3 (or a variant of SEQ ID NO: 3 with one or two amino acid modifications), and / or (b) a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 9. The binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers and / or ADCs) provided herein having a light chain variable domain having a CDR1 having the amino acid sequence of SEQ ID NO: 9 (or a variant of SEQ ID NO: 9 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO: 10 (or a variant of SEQ ID NO: 10 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO: 11 (or a variant of SEQ ID NO: 11 with one or two amino acid modifications), can comprise any suitable framework region.For example, such binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers and / or ADCs) may comprise: (a) a framework region 1 having the amino acid sequence set forth in SEQ ID NO:4 (or a variant of SEQ ID NO:4 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications); framework region 2 having the amino acid sequence set forth in SEQ ID NO: 6 (or a variant of SEQ ID NO: 6 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications); framework region 3 having the amino acid sequence set forth in SEQ ID NO: 7 (or a variant of SEQ ID NO: 7 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications); and / or (b) a heavy chain variable domain comprising framework region 4 having the amino acid sequence set forth in SEQ ID NO: 12 (or a variant of SEQ ID NO: 12 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), framework region 1 having the amino acid sequence set forth in SEQ ID NO: 13 (or a variant of SEQ ID NO: 13 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications). and a light chain variable domain comprising framework region 2 having the amino acid sequence set forth in SEQ ID NO: 14 (or a variant of SEQ ID NO: 14 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), and framework region 4 having the amino acid sequence set forth in SEQ ID NO: 15 (or a variant of SEQ ID NO: 15 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications).

[0090] In some cases, a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) having any of the CDRs set forth in Figures 2A or 2B may be designed to include the framework regions set forth in Figures 2A and 2B, or may be designed to include one or more framework regions from another antibody, antibody fragment or antibody domain. For example, a Fab may be designed to include the six CDRs set forth in Figures 2A and 2B and the framework regions set forth in Figures 2A and 2B, except that framework region 1 having the amino acids set forth in SEQ ID NO:4 is replaced with framework region 1 having the amino acids set forth in SEQ ID NO:20, or framework region 1 having the amino acids set forth in SEQ ID NO:54, or framework region 1 having the amino acids set forth in SEQ ID NO:64. In some cases, a scFv may be designed to include the six CDRs set forth in Figures 2A and 2B and the framework regions set forth in Figures 2A and 2B. In some cases, an scFv can be designed to include the six CDRs set forth in Figures 2A and 2B and the framework regions set forth in Figures 2A and 2B, except that framework region 1 having the amino acids set forth in SEQ ID NO: 4 is replaced with framework region 1 having the amino acids set forth in SEQ ID NO: 20, framework region 1 having the amino acids set forth in SEQ ID NO: 54, or framework region 1 having the amino acids set forth in SEQ ID NO: 64. In another example, an scFv can be designed to include the six CDRs set forth in Figures 2A and 2B and the framework regions set forth in Figures 6B and 6C, Figures 7A and 7B, Figures 8A and 8B, or Figures 9A and 9B.

[0091] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise (a) a heavy chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8, and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16. For example, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) can comprise (a) a heavy chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:8, and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:16. In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) can comprise (a) a heavy chain variable domain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:8, and / or (b) a light chain variable domain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:16.

[0092] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise: (a) a heavy chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8, with the proviso that it comprises the amino acid sequence set forth in SEQ ID NOs:1, 2, and 3; and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16, with the proviso that it comprises the amino acid sequence set forth in SEQ ID NOs:9, 10, and 11. For example, a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) provided herein may comprise: (a) a heavy chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:8, with the proviso that the heavy chain variable domain comprises an amino acid sequence set forth in SEQ ID NO:1, 2, and 3; and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:16, with the proviso that the light chain variable domain comprises an amino acid sequence set forth in SEQ ID NO:9, 10, and 11.

[0093] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) having the ability of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise (a) a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 8, or the amino acids set forth in SEQ ID NO: 8 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), and / or (b) a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 16, or the amino acids set forth in SEQ ID NO: 16 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). For example, the antibodies or antigen-binding fragments provided herein may have the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and may comprise a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 8 with one, two, three, four, five, six, seven, eight, nine or ten amino acid modifications (e.g., amino acid substitutions, amino acid deletions and / or amino acid additions), provided that they comprise the amino acid sequences set forth in SEQ ID NOs: 1, 2 and 3, and may comprise a light chain variable domain having the amino acid sequence set forth in SEQ ID NO: 16 with one, two, three, four, five, six, seven, eight, nine or ten amino acid modifications (e.g., amino acid substitutions, amino acid deletions and / or amino acid additions), provided that they comprise the amino acid sequences set forth in SEQ ID NOs: 9, 10 and 11.

[0094] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise: (a) a heavy chain variable domain comprising: (i) a CDR1 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO:1; (ii) a CDR2 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO:2; and (iii) a CDR3 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO:3; and / or (b) a light chain variable domain comprising: (i) a CDR1 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO:9; (ii) a CDR2 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO:10; and (iii) a CDR3 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO:11. As used herein, a "CDR1 consisting essentially of the amino acid sequence set forth in SEQ ID NO:1" is a CDR1 having 0, 1, or 2 amino acid substitutions within SEQ ID NO:1, having 0, 1, 2, 3, 4, or 5 amino acid residues immediately preceding SEQ ID NO:1, and / or having 0, 1, 2, 3, 4, or 5 amino acid residues immediately following SEQ ID NO:1, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR1s consisting essentially of the amino acid sequence set forth in SEQ ID NO:1 include, but are not limited to, those set forth in Table 1.

[0095] [Table 1]

[0096] As used herein, a "CDR2 consisting essentially of the amino acid sequence set forth in SEQ ID NO:2" is a CDR2 that has 0, 1, or 2 amino acid substitutions within SEQ ID NO:2, has 0, 1, 2, 3, 4, or 5 amino acid residues immediately preceding SEQ ID NO:2, and / or has 0, 1, 2, 3, 4, or 5 amino acid residues immediately following SEQ ID NO:2, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR2s consisting essentially of the amino acid sequence set forth in SEQ ID NO:2 include, but are not limited to, those set forth in Table 2.

[0097] [Table 2]

[0098] As used herein, a "CDR3 consisting essentially of the amino acid sequence set forth in SEQ ID NO:3" is a CDR3 that has 0, 1, or 2 amino acid substitutions within SEQ ID NO:3, has 0, 1, 2, 3, 4, or 5 amino acid residues immediately preceding SEQ ID NO:3, and / or has 0, 1, 2, 3, 4, or 5 amino acid residues immediately following SEQ ID NO:3, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR3s consisting essentially of the amino acid sequence set forth in SEQ ID NO:3 include, but are not limited to, those set forth in Table 3.

[0099] [Table 3]

[0100] As used herein, a "CDR1 consisting essentially of the amino acid sequence set forth in SEQ ID NO:9" is a CDR1 having zero, one or two amino acid substitutions within SEQ ID NO:9, having zero, one, two, three, four or five amino acid residues immediately preceding SEQ ID NO:9, and / or having zero, one, two, three, four or five amino acid residues immediately following SEQ ID NO:9, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR1s consisting essentially of the amino acid sequence set forth in SEQ ID NO:9 include, but are not limited to, those set forth in Table 4.

[0101] [Table 4]

[0102] As used herein, a "CDR2 consisting essentially of the amino acid sequence set forth in SEQ ID NO: 10" is a CDR2 that has zero, one or two amino acid substitutions within SEQ ID NO: 10, has zero, one, two, three, four or five amino acid residues immediately preceding SEQ ID NO: 10, and / or has zero, one, two, three, four or five amino acid residues immediately following SEQ ID NO: 10, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR2s consisting essentially of the amino acid sequence set forth in SEQ ID NO: 10 include, but are not limited to, those set forth in Table 5.

[0103] [Table 5]

[0104] As used herein, a "CDR3 consisting essentially of the amino acid sequence set forth in SEQ ID NO:11" is a CDR3 that has 0, 1, or 2 amino acid substitutions within SEQ ID NO:11, has 0, 1, 2, 3, 4, or 5 amino acid residues immediately preceding SEQ ID NO:11, and / or has 0, 1, 2, 3, 4, or 5 amino acid residues immediately following SEQ ID NO:11, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR3s consisting essentially of the amino acid sequence set forth in SEQ ID NO:11 include, but are not limited to, those set forth in Table 6.

[0105] [Table 6]

[0106] In another embodiment, a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) provided herein having the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) comprises (i) a CDR1 having the amino acid sequence set forth in SEQ ID NO: 17 (or a variant of SEQ ID NO: 17 having one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO: 18 (or a variant of SEQ ID NO: 18 having one or two amino acid modifications), and an amino acid sequence set forth in SEQ ID NO: 19 (or and / or (ii) a light chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:25 (or a variant of SEQ ID NO:25 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:26 (or a variant of SEQ ID NO:26 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:27 (or a variant of SEQ ID NO:27 with one or two amino acid modifications). Examples of such antigen-binding fragments having these CDRs and the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) include, but are not limited to, the Fabs set forth in Figures 3A and 3B.

[0107] In some cases, the antibody has the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and has (a) a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO: 17 (or a variant of SEQ ID NO: 17 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO: 18 (or a variant of SEQ ID NO: 18 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO: 19 (or a variant of SEQ ID NO: 19 with one or two amino acid modifications), and / or (b) a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO: 25 (or a variant of SEQ ID NO: 25 with one or two amino acid modifications). The binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers and / or ADCs) provided herein having a light chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO: 25 (or a variant of SEQ ID NO: 25 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO: 26 (or a variant of SEQ ID NO: 26 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO: 27 (or a variant of SEQ ID NO: 27 with one or two amino acid modifications), can comprise any suitable framework region.For example, such a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) may comprise: (a) a framework region 1 having the amino acid sequence set forth in SEQ ID NO:20 (or a variant of SEQ ID NO:20 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), an amino acid sequence set forth in SEQ ID NO:21 (or a variant of SEQ ID NO:21 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), and / or a heavy chain variable domain comprising framework region 2 having the amino acid sequence set forth in SEQ ID NO: 22 (or a variant of SEQ ID NO: 22 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), framework region 3 having the amino acid sequence set forth in SEQ ID NO: 22 (or a variant of SEQ ID NO: 22 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications). or (b) a light chain variable domain comprising a framework region 1 having the amino acid sequence set forth in SEQ ID NO:28 (or a variant of SEQ ID NO:28 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:29 (or a variant of SEQ ID NO:29 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:30 (or a variant of SEQ ID NO:30 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:31 (or a variant of SEQ ID NO:31 having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid modifications).

[0108] In some cases, a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) having any of the CDRs set forth in Figures 3A or 3B may be designed to include the framework regions set forth in Figures 3A and 3B, or may be designed to include one or more framework regions from another antibody or antibody fragment. For example, a Fab may be designed to include the six CDRs set forth in Figures 3A and 3B and the framework regions set forth in Figures 3A and 3B, except that framework region 1 having the amino acids set forth in SEQ ID NO:20 is replaced with framework region 1 having the amino acids set forth in SEQ ID NO:4, or framework region 1 having the amino acids set forth in SEQ ID NO:54, or framework region 1 having the amino acids set forth in SEQ ID NO:64. In some cases, a scFv may be designed to include the six CDRs set forth in Figures 3A and 3B and the framework regions set forth in Figures 3A and 3B. In some cases, an scFv can be designed to include the six CDRs set forth in Figures 3A and 3B and the framework regions set forth in Figures 3A and 3B, except that framework region 1 having the amino acids set forth in SEQ ID NO: 20 is replaced with framework region 1 having the amino acids set forth in SEQ ID NO: 4, framework region 1 having the amino acids set forth in SEQ ID NO: 54, or framework region 1 having the amino acids set forth in SEQ ID NO: 64. In another example, an scFv can be designed to include the six CDRs set forth in Figures 3A and 3B and the framework regions set forth in Figures 6B and 6C, Figures 7A and 7B, Figures 8A and 8B, or Figures 9A and 9B.

[0109] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise (a) a heavy chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24, and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32. For example, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) can comprise (a) a heavy chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 24, and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 32. In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) can comprise (a) a heavy chain variable domain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 24, and / or (b) a light chain variable domain comprising an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO: 32.

[0110] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise: (a) a heavy chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 24, with the proviso that it comprises the amino acid sequence set forth in SEQ ID NO: 17, 18, and 19; and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO: 32, with the proviso that it comprises the amino acid sequence set forth in SEQ ID NO: 25, 26, and 27. For example, a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) provided herein may comprise: (a) a heavy chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 24, with the proviso that the heavy chain variable domain comprises an amino acid sequence set forth in SEQ ID NO: 17, 18, and 19; and / or (b) a light chain variable domain comprising an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO: 32, with the proviso that the light chain variable domain comprises an amino acid sequence set forth in SEQ ID NO: 25, 26, and 27.

[0111] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) having the ability of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise (a) a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 24, or the amino acids set forth in SEQ ID NO: 24 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), and / or (b) a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 32, or the amino acids set forth in SEQ ID NO: 32 with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions). For example, the antibodies or antigen-binding fragments provided herein may have the ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and may comprise a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO: 24 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that they comprise the amino acid sequences set forth in SEQ ID NOs: 17, 18, and 19; and may comprise a light chain variable domain having the amino acid sequence set forth in SEQ ID NO: 32 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and / or amino acid additions), provided that they comprise the amino acid sequences set forth in SEQ ID NOs: 25, 26, and 27.

[0112] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) may comprise: (a) a heavy chain variable domain comprising: (i) a CDR1 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO: 17; (ii) a CDR2 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO: 18; and (iii) a CDR3 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO: 19; and / or (b) a light chain variable domain comprising: (i) a CDR1 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO: 25; (ii) a CDR2 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO: 26; and (iii) a CDR3 comprising, consisting essentially of, or consisting of the amino acid sequence set forth in SEQ ID NO: 27. As used herein, a "CDR1 consisting essentially of the amino acid sequence set forth in SEQ ID NO: 17" is a CDR1 having 0, 1, or 2 amino acid substitutions within SEQ ID NO: 17, having 0, 1, 2, 3, 4, or 5 amino acid residues immediately preceding SEQ ID NO: 17, and / or having 0, 1, 2, 3, 4, or 5 amino acid residues immediately following SEQ ID NO: 17, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR1s consisting essentially of the amino acid sequence set forth in SEQ ID NO: 17 include, but are not limited to, those set forth in Table 7.

[0113] [Table 7]

[0114] As used herein, a "CDR2 consisting essentially of the amino acid sequence set forth in SEQ ID NO: 18" is a CDR2 that has zero, one or two amino acid substitutions within SEQ ID NO: 18, has zero, one, two, three, four or five amino acid residues immediately preceding SEQ ID NO: 18, and / or has zero, one, two, three, four or five amino acid residues immediately following SEQ ID NO: 18, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR2s consisting essentially of the amino acid sequence set forth in SEQ ID NO: 18 include, but are not limited to, those set forth in Table 8.

[0115] [Table 8]

[0116] As used herein, a "CDR3 consisting essentially of the amino acid sequence set forth in SEQ ID NO: 19" is a CDR3 that has zero, one or two amino acid substitutions within SEQ ID NO: 19, has zero, one, two, three, four or five amino acid residues immediately preceding SEQ ID NO: 19, and / or has zero, one, two, three, four or five amino acid residues immediately following SEQ ID NO: 19, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR3s consisting essentially of the amino acid sequence set forth in SEQ ID NO: 19 include, but are not limited to, those set forth in Table 9.

[0117] [Table 9]

[0118] As used herein, a "CDR1 consisting essentially of the amino acid sequence set forth in SEQ ID NO:25" is a CDR1 having 0, 1 or 2 amino acid substitutions within SEQ ID NO:25, having 0, 1, 2, 3, 4 or 5 amino acid residues immediately preceding SEQ ID NO:25, and / or having 0, 1, 2, 3, 4 or 5 amino acid residues immediately following SEQ ID NO:25, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR1s consisting essentially of the amino acid sequence set forth in SEQ ID NO:25 include, but are not limited to, those listed in Table 10.

[0119] [Table 10]

[0120] As used herein, a "CDR2 consisting essentially of the amino acid sequence set forth in SEQ ID NO:26" is a CDR2 that has zero, one or two amino acid substitutions within SEQ ID NO:26, has zero, one, two, three, four or five amino acid residues immediately preceding SEQ ID NO:26, and / or has zero, one, two, three, four or five amino acid residues immediately following SEQ ID NO:26, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR2s consisting essentially of the amino acid sequence set forth in SEQ ID NO:26 include, but are not limited to, those set forth in Table 11.

[0121] [Table 11]

[0122] As used herein, a "CDR3 consisting essentially of the amino acid sequence set forth in SEQ ID NO:27" is a CDR3 that has zero, one or two amino acid substitutions within SEQ ID NO:27, has zero, one, two, three, four or five amino acid residues immediately preceding SEQ ID NO:27, and / or has zero, one, two, three, four or five amino acid residues immediately following SEQ ID NO:27, provided that the binding agent (e.g., antibody, antigen-binding fragment, antibody domain, CAR, cell engager and / or ADC) maintains the basic ability to bind to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of CDR3s consisting essentially of the amino acid sequence set forth in SEQ ID NO:27 include, but are not limited to, those set forth in Table 12.

[0123] [Table 12]

[0124] When designing a single chain antibody (e.g., scFv) having a heavy chain variable domain and a light chain variable domain, the two regions may be directly linked or may be linked using any suitable linker sequence. For example, a heavy chain variable domain having CDRs of SEQ ID NOs: 1 to 3 or 17 to 19 may be directly linked via a linker sequence to a light chain variable domain having CDRs of SEQ ID NOs: 9 to 11 or 25 to 27, respectively. Examples of linker sequences that can be used to link the heavy chain variable domain and the light chain variable domain to create an scFv include, but are not limited to, the linkers shown in FIG. 10.

[0125] As indicated herein, the amino acid sequences described herein may include amino acid modifications (e.g., a concatenated number of amino acid modifications). Such amino acid modifications may include, but are not limited to, amino acid substitutions, amino acid deletions, amino acid additions, and combinations. In some cases, amino acid modifications may be made to improve binding and / or contact with an antigen and / or to improve the functional activity of a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) provided herein. In some cases, the amino acid substitutions in the concatenated sequence identifiers may be conservative amino acid substitutions. For example, a conservative amino acid substitution may be made by replacing one amino acid residue with another amino acid residue having a similar side chain. A family of amino acid residues having similar side chains can include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0126] In some cases, the amino acid substitutions in the concatenated sequence identifiers may be non-conservative amino acid substitutions. Non-conservative amino acid substitutions can be made by replacing one amino acid residue with another amino acid residue having a dissimilar side chain. Examples of non-conservative substitutions include, but are not limited to, replacing (a) a hydrophilic residue (e.g., serine or threonine) with a hydrophobic residue (e.g., leucine, isoleucine, phenylalanine, valine, or alanine), (b) cysteine ​​or proline with any other residue, (c) a residue having a basic side chain (e.g., lysine, arginine, or histidine) with a residue having an acidic side chain (e.g., aspartic acid or glutamic acid), and (d) a residue having a bulky side chain (e.g., phenylalanine) with glycine or other residues having small side chains.

[0127] Methods for generating amino acid sequence variants (e.g., amino acid sequences containing one or more modifications relative to the concatenated sequence identifiers) can include site-directed or random mutagenesis (e.g., by PCR) of a nucleic acid encoding an antibody or fragment thereof. See, e.g., Zoller, Curr. Opin. Biotechnol. 3: 348-354 (1992). Both naturally occurring and non-naturally occurring amino acids (e.g., artificially derivatized amino acids) can be used to generate the amino acid sequence variants provided herein.

[0128] Further set forth in Table 13 are representative numbers of binding agents (eg, antibodies, antigen-binding fragments, and / or antibody domains) capable of binding to a CD94 / NKG2A polypeptide (eg, a human CD94 / NKG2A polypeptide).

[0129] [Table 13]

[0130] [Table 14]

[0131] The binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) can be produced using any suitable method. For example, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers) can be produced in recombinant host cells. For example, nucleic acids encoding the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers) can be constructed, introduced into an expression vector, and expressed in a suitable host cell. Figure 4 is a sequence listing of nucleic acid sequences encoding exemplary binding agents (e.g., antibodies, antigen-binding fragments, and / or antibody domains) described herein. In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, and / or cell engager) can be recombinantly produced in a prokaryotic host, such as E. coli, Bacillus brevis, Bacillus subtilis, Bacillus megaterium, Lactobacillus zeae / casei, or Lactobacillus paracasei.The binding agents (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers) provided herein can be expressed in eukaryotic hosts, such as yeast (e.g., Pichia pastoris, Saccharomyces cerevisiae, Hansenula polymorpha, Schizosaccharomyces pombe, Schwanniomyces occidentalis, Kluyveromyces lactis, or Yarrowia lipolytica), filamentous fungi of the genus Trichoderma (e.g., T. reesei), and Aspergillus (e.g., A. niger). The antibodies can also be recombinantly produced in A. niger and A. oryzae, protozoa such as Leishmania tarentolae, insect cells, or mammalian cells (e.g., mammalian cell lines such as Chinese hamster ovary (CHO) cells, Per.C6 cells, mouse myeloma NS0 cells, baby hamster kidney (BHK) cells, or the human embryonic kidney cell line HEK293). See, e.g., Frenzel et al., Front Immunol., 4:217 (2013).

[0132] In some cases, the antigen-binding fragments or antibody domains provided herein can be produced by proteolytic digestion of intact antibodies. For example, antigen-binding fragments can be obtained by treating antibodies with enzymes such as papain or pepsin. Papain digestion of whole antibodies can be used to produce F(ab)2 or Fab fragments, while pepsin digestion of whole antibodies can be used to produce F(ab')2 or Fab' fragments.

[0133] In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) may be substantially pure. The term "substantially pure" as used herein with respect to a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) refers to a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) that is substantially free of other polypeptides, lipids, carbohydrates, and nucleic acids with which it is naturally associated. That is, a substantially pure binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) is any binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) that is removed from its natural environment and is at least 60% pure. The substantially pure binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) may be at least about 65, 70, 75, 80, 85, 90, 95, or 99 percent pure.

[0134] This document also provides bispecific binding agents (e.g., bispecific antibodies, bispecific antigen-binding fragments, and / or bispecific antibody domains) that bind to two different epitopes, at least one of which is an epitope of a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). In some cases, the bispecific binding agents provided herein can be designed to bind to two different epitopes of the same CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). In some cases, the bispecific binding agents provided herein can bind to an epitope on a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and a different polypeptide (e.g., a CD3 polypeptide). Bispecific binding agents can be produced by chemically conjugating two different binding agents (e.g., antibodies, antigen-binding fragments, and / or antibody domains) together. Bispecific binding agents can also be produced by fusing two antibody producing cells, e.g., hybridomas, to create a hybrid cell line producing two different heavy chains and two different light chains in the same cell, thereby resulting in, e.g., a bispecific IgG molecule. See Brinkmann and Kontermann, MAbs., 9(2):182-212 (2017).

[0135] In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers) can be fused or conjugated (e.g., covalently or non-covalently linked) to another polypeptide or other moiety to obtain a fusion protein or conjugate. For example, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers) can be conjugated (e.g., covalently or non-covalently linked) to a polymer (e.g., polyethylene glycol (PEG), PEG-modified polyethyleneimine (PEI) (PEI-PEG), and / or polyglutamic acid (PGA) (N-(2-hydroxypropyl) methacrylamide (HPMA) copolymers), hyaluronic acid, a fluorescent agent, a luminescent agent, a hapten, an enzyme, a metal chelate, a drug, a radioisotope, and / or a cytotoxic agent. Any suitable method may be used to conjugate (e.g., covalently or non-covalently link) another polypeptide or other moiety to the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers). For example, the methods described in U.S. Pat. No. 8,021,661 can be used to conjugate another polypeptide or other moiety to the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers).

[0136] In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) can be modified with a moiety that improves stabilization and / or retention in circulation, e.g., in blood, serum, or other tissues, e.g., at least 1.5-fold, 2-fold, 5-fold, 10-fold, or 50-fold. For example, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) can be conjugated (e.g., covalently or non-covalently conjugated) to a polymer, e.g., a substantially non-antigenic polymer. Examples of substantially non-antigenic polymers that can be used as described herein include, but are not limited to, polyalkylene oxides and polyethylene oxides. In some cases, the polymers used herein can have any suitable molecular weight. For example, polymers having an average molecular weight of about 200 daltons to about 35,000 daltons (e.g., about 1,000 to about 15,000 daltons or about 2,000 to about 12,500 daltons) can be used. In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, cell engagers, and / or ADCs) can be conjugated (e.g., covalently or non-covalently conjugated) to a water-soluble polymer. Examples of water-soluble polymers that can be used as described herein include, but are not limited to, hydrophilic polyvinyl polymers, polyvinyl alcohol, polyvinylpyrrolidone, polyalkylene oxide homopolymers, polyethylene glycol (PEG), polypropylene glycol, polyoxyethylenated polyols, and copolymers and / or block copolymers thereof, provided that the water solubility of the copolymer or block copolymer is maintained.

[0137] In some cases, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) can be conjugated (e.g., covalently or non-covalently bound) to one or more polyoxyalkylenes (e.g., polyoxyethylene, polyoxypropylene, or block copolymers of polyoxyethylene and polyoxypropylene), polymethacrylates, carbomers, branched or unbranched polysaccharides, or combinations thereof. For example, a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) can be covalently bound to polyoxyethylene.

[0138] This document also provides ADCs. As used herein, the term "ADC" refers to a conjugate comprising (a) an antigen-binding domain and (b) at least one drug directly or indirectly covalently linked to the antigen-binding domain. In some cases, the ADCs described herein may comprise (a) an antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) and (b) at least one drug directly or indirectly covalently linked to the antigen-binding domain. Any suitable binding agent (e.g., an antibody, an antigen-binding fragment, and / or an antibody domain) provided herein and capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) can be used as an antigen-binding domain to generate the ADCs described herein. For example, any of the binding agents listed in Table 13 can be used to generate ADCs capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide). Examples of drugs that can be used to make the ADCs described herein include, but are not limited to, small molecule inhibitors of IL-2, IL-12, and PD-L1 (e.g., BMS1166 or BMS202). In some cases, examples of drugs that can be used to make the ADCs described herein include, but are not limited to, auristatins (e.g., monomethylauristatin E (MMAE)), mertansine (DM-1), and pyrrolobenzodiazepine (PBD) dimers. Any suitable ADC linker can be used to covalently link one or more drugs to an antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) to form the ADCs provided herein. For example, a cleavable or non-cleavable ADC linker can be used to covalently link one or more drugs to an antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) to form the ADCs provided herein.Examples of ADC linkers that can be used to covalently attach one or more drugs to an antigen-binding domain capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) to form an ADC provided herein include, but are not limited to, an ADC disulfide linker, an ADC hydrazone linker, an ADC peptide linker, an ADC thioether linker, and an ADC PEG-containing linker.

[0139] This document also provides a nucleic acid molecule (e.g., an isolated nucleic acid molecule) having a nucleic acid sequence encoding at least a portion of a binding agent (e.g., an antibody, an antigen-binding fragment, an antibody domain, a CAR, and / or a cell engager) provided herein. For example, an isolated nucleic acid molecule provided herein can include a nucleic acid sequence encoding a heavy chain variable domain, e.g., a heavy chain variable domain as set forth in FIG. 2A or 3A. In another example, an isolated nucleic acid molecule provided herein can include a nucleic acid sequence encoding a light chain variable domain, e.g., a light chain variable domain as set forth in FIG. 2B or 3B. In some cases, an isolated nucleic acid molecule provided herein can include a nucleic acid sequence encoding both (a) a heavy chain light chain variable domain and (b) a light chain variable domain, with or without encoding a linker polypeptide as set forth in FIG. 10. A nucleic acid (e.g., an isolated nucleic acid molecule) provided herein can be a single-stranded or double-stranded nucleic acid of any suitable type (e.g., DNA, RNA, or DNA / RNA hybrid).

[0140] This document also provides vectors (e.g., plasmid vectors or viral vectors) that contain one or more nucleic acids provided herein. Examples of plasmid vectors that can be designed to contain one or more nucleic acids having a nucleic acid sequence encoding at least a portion of the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers) include, but are not limited to, phagemids. Examples of viral vectors that can be designed to contain one or more nucleic acids having a nucleic acid sequence encoding at least a portion of the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers) include, but are not limited to, retroviral vectors, parvoviral vectors (e.g., adenoviral vectors and adeno-associated virus (AAV) vectors), lentiviral vectors (e.g., herpes simplex (HSV) vectors), poxviral vectors (e.g., vaccinia virus vectors and fowlpox virus vectors), and hybrid or chimeric viral vectors. For example, viral vectors having lentiviral components, such as adenoviral backbones including those described elsewhere (Zheng et al., Nat. Biotech., 18(2): 176-80 (2000); WO 98 / 22143; WO 98 / 46778; and WO 00 / 17376), or AAV components, such as adenoviral backbones including those described elsewhere (Fisher et al., Hum. Gene Ther., 7:2079-2087 (1996)), can be engineered to include one or more nucleic acids having a nucleic acid sequence encoding at least a portion of a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, and / or cell engager).

[0141] In some cases, a vector provided herein (e.g., a plasmid vector or a viral vector) can include a nucleic acid sequence encoding an scFv or an antibody domain (e.g., a VH domain) provided herein. In some cases, a vector provided herein (e.g., a plasmid vector or a viral vector) can include a nucleic acid sequence encoding a CAR provided herein. In some cases, a vector provided herein (e.g., a plasmid vector or a viral vector) can include a nucleic acid sequence encoding a cell engager provided herein.

[0142] The vectors provided herein (e.g., plasmid vectors or viral vectors provided herein) can include any suitable promoter and other control sequences (e.g., transcription and translation start and stop codons) operably linked to a nucleic acid sequence encoding at least a portion of a binding agent provided herein (e.g., an antibody, an antigen-binding fragment, an antibody domain, a CAR, and / or a cell engager). In some cases, the promoter used to drive expression can be a constitutive promoter or a controllable promoter. Examples of controllable promoters that can be used as described herein include, but are not limited to, inducible promoters, repressible promoters, and tissue-specific promoters. Examples of viral promoters that can be used as described herein include, but are not limited to, adenovirus promoters, vaccinia virus promoters, CMV promoters (e.g., immediate early CMV promoters), and AAV promoters.

[0143] Any suitable method can be used to generate a nucleic acid molecule (or vector, e.g., a plasmid vector or a viral vector) having a nucleic acid sequence encoding at least a portion of a binding agent (e.g., an antibody, an antigen-binding fragment, an antibody domain, a CAR, and / or a cell engager) provided herein. For example, molecular cloning techniques can be used to generate a nucleic acid molecule (or vector, e.g., a plasmid vector or a viral vector) having a nucleic acid sequence encoding at least a portion of a binding agent (e.g., an antibody, an antigen-binding fragment, an antibody domain, a CAR, and / or a cell engager) provided herein, as described elsewhere (see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, NY (1989); and Ausubel et al., Current Protocols in Molecular Biology, Green Publishing Associates and John Wiley & Sons, New York, NY (1994)).

[0144] This document also provides host cells that include a nucleic acid provided herein (e.g., a nucleic acid having a nucleic acid sequence encoding at least a portion of a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, and / or cell engager). Host cells that can be engineered to include one or more nucleic acids provided herein can be prokaryotic or eukaryotic cells. Examples of prokaryotic cells that can be engineered to include a nucleic acid provided herein include, but are not limited to, E. coli (e.g., Tb-1, TG-1, DH5α, XL-Blue MRF (Stratagene), SA2821, or Y1090 cells), Bacillus subtilis, Salmonella typhimurium, Serratia marcescens, or Pseudomonas (e.g., P. aerugenosa) cells. Examples of eukaryotic cells that can be engineered to contain a nucleic acid provided herein include, but are not limited to, insect cells (e.g., Sf9 or Ea4 cells), yeast cells (e.g., S. cerevisiae cells), and mammalian cells (e.g., mouse, rat, hamster, monkey, or human cells). For example, VERO cells, HeLa cells, 3T3 cells, Chinese hamster ovary (CHO) cells, W138 BHK cells, COS-7 cells, and MDCK cells can be engineered to contain a nucleic acid provided herein. Any suitable method may be used to introduce one or more nucleic acids provided herein (e.g., a vector, such as a plasmid vector or a viral vector, having a nucleic acid sequence encoding at least a portion of a binding agent provided herein) into a host cell. For example, calcium chloride-mediated transformation, transduction, conjugation, triparental mating, DEAE, dextran-mediated transfection, infection, membrane fusion with liposomes, high velocity bombardment with DNA-coated microprojectiles, direct microinjection into single cells, electroporation, or a combination thereof can be used to introduce the nucleic acids provided herein into a host cell (see, e.g., Sambrook et al., Molecular Biology: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1999). Harbor Laboratory, NY (1989); Davis et al., Basic Methods in Molecular Biology (1986); and Neumann et al., EMBO J., 1:841 (1982).

[0145] In some cases, cells, such as T cells, stem cells (e.g., induced pluripotent stem cells or mesenchymal stem cells), or NK cells, can be engineered to express one or more nucleic acids encoding a CAR described herein. For example, a population of T cells can be infected with a viral vector designed to express a nucleic acid encoding a CAR described herein (e.g., a CAR having the ability to bind to a CD94 / NKG2A polypeptide).

[0146] In some cases, cells, such as T cells, stem cells (e.g., induced pluripotent stem cells or mesenchymal stem cells), or NK cells, can be engineered to express one or more nucleic acids encoding a cell engager described herein. For example, a population of T cells can be infected with a viral vector designed to express a nucleic acid encoding a cell engager described herein (e.g., a cell engager capable of binding to a CD94 / NKG2A polypeptide).

[0147] In some cases, a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, and / or cell engager) provided herein can be produced using a method that includes the steps of (a) introducing a nucleic acid encoding a polypeptide into a host cell, (b) culturing the host cell in a culture medium under conditions sufficient to express the polypeptide, (c) recovering the polypeptide from the cell or the culture medium, and (d) purifying the polypeptide (e.g., to reach at least 50, 60, 70, 80, 90, 95, 97, 98, or 99 percent purity).

[0148] In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, cell engagers, and / or ADCs), nucleic acids provided herein (e.g., nucleic acids encoding antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers provided herein), vectors provided herein (e.g., viral vectors designed to express antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers provided herein), and / or host cells provided herein (e.g., host cells designed to express antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers provided herein) can be formulated as pharmaceutical compositions for administration to a mammal (e.g., a human) with cancer to treat the mammal. In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, cell engagers, and / or ADCs), nucleic acids provided herein (e.g., nucleic acids encoding antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers provided herein), vectors provided herein (e.g., viral vectors designed to express antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers provided herein), and / or host cells provided herein (e.g., host cells designed to express antibodies, antigen-binding fragments, antibody domains, CARs, and / or cell engagers provided herein) can be formulated as pharmaceutical compositions for administration to a mammal (e.g., a human) to reduce the number of cancer cells in the mammal and / or increase the survival of the mammal afflicted with cancer. For example, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, antibody domains, cell engagers, and / or ADCs) capable of binding to a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) can be formulated as pharmaceutical compositions for administration to a mammal (e.g., a human).In some cases, the pharmaceutical compositions provided herein may include a pharma- ceutically acceptable carrier, such as a buffer, salt, surfactant, sugar, tonicity modifier, or combinations thereof, as described elsewhere (Gervasi et al., Eur. J. Pharmaceutics and Biopharmaceutics, 131:8-24 (2018)). Examples of pharma-ceutically acceptable carriers that can be used to make the pharmaceutical compositions provided herein include, but are not limited to, water, lactic acid, citric acid, sodium chloride, sodium citrate, sodium succinate, sodium phosphate, surfactants (e.g., polysorbate 20, polysorbate 80, or poloxamer 188), dextran 40, or sugars (e.g., sorbitol, mannitol, sucrose, dextrose, or trehalose), or combinations thereof. For example, a pharmaceutical composition designed to include a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, cell engager, and / or ADC) (or a nucleic acid, vector, or host cell provided herein) can be formulated to include a buffer (e.g., acetate, citrate, histidine, succinate, phosphate, or hydroxymethylaminomethane (Tris) buffer), a surfactant (e.g., polysorbate 20, polysorbate 80, or poloxamer 188), and a sugar, such as sucrose. Other components that can be included in the pharmaceutical compositions provided herein include, but are not limited to, amino acids such as glycine or arginine, antioxidants such as ascorbic acid, methionine, or ethylenediaminetetraacetic acid (EDTA), anticancer drugs such as enzalutamide, imatinib, gefitinib, erlotinib, sunitinib, lapatinib, nilotinib, sorafenib, temsirolimus, everolimus, pazopanib, crizotinib, ruxolitinib, axitinib, bosutinib, cabozantinib, ponatinib, regorafenib, ibrutinib, trametinib, perifosine, bortezomib, carfilzomib, batimastat, ganetespib, obatoclax, navitoclax, taxol, paclitaxel, or bevacizumab, or combinations thereof.For example, the pharmaceutical compositions provided herein can be used in combination with one or more checkpoint inhibitors, such as an anti-PD-1 antibody or PD-1 inhibitor (e.g., cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, or AMP-514), an anti-PD-L1 antibody or PD-L1 inhibitor (e.g., avelumab, durvalumab, atezolizumab, KN033, KN034, KN036, KN038, KN039 ... The present invention can be formulated to include one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cells engineered to express a CAR capable of binding to a CD94 / NKG2A polypeptide, one or more cell engagers, and / or one or more ADCs) in combination with an anti-CTLA-4 antibody (e.g., ipilimumab), an anti-CTLA-4 antibody (e.g., ipilimumab), or an anti-IL-1 antibody (e.g., ipilimumab).

[0149] In some cases, when formulating a pharmaceutical composition to include one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cells engineered to express a CAR capable of binding to a CD94 / NKG2A polypeptide, one or more cell engagers, and / or one or more ADCs), any suitable concentration of the binding agent can be used. For example, the pharmaceutical compositions provided herein contain about 1 mg to about 500 mg (e.g., about 1 mg to about 500 mg, about 10 mg to about 500 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 0.5 mg to about 250 mg, about 0.5 mg to about 150 mg, about 0.5 mg to about 100 mg, about 0.5 mg to about 50 mg, about 1 mg to about 300 mg, about 2 mg to about 200 mg, about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 150 mg, or about 150 mg to about 300 mg) of a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, CAR +In another example, the pharmaceutical compositions provided herein may be formulated to be a solid or semi-solid containing about 0.5 mg to about 500 mg (e.g., about 1 mg to about 500 mg, about 10 mg to about 500 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 0.5 mg to about 250 mg, about 0.5 mg to about 150 mg, about 0.5 mg to about 100 mg, about 0.5 mg to about 50 mg, about 1 mg to about 300 mg, about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 150 mg, or about 150 mg to about 300 mg) of a binding agent provided herein (e.g., an antibody, antigen-binding fragment, antibody domain, cell engager, and / or ADC). In some cases, pharmaceutical compositions containing a binding agent (e.g., an antibody, antigen-binding fragment, and / or antibody domain) provided herein may be used to treat a patient with a binding agent titer of about 1×10 5 ~Approx. 1×10 12 (For example, about 1×10 5 ~Approx. 1×10 10 , about 1×10 5 ~Approx. 1×10 8 , about 1×10 6 ~Approx. 1×10 12 , about 1×10 6 ~Approx. 1×10 12 , about 1×10 8 ~Approx. 1×10 12 , about 1×10 9 ~Approx. 1×10 12 , about 1×10 6 ~Approx. 1×10 11 , or about 1 × 10 7 ~Approx. 1×10 10 ) may be formulated as a dosage form.

[0150] In some cases, when a pharmaceutical composition is formulated to include one or more nucleic acids (e.g., a vector, e.g., a viral vector) encoding at least a portion of a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, CAR, and / or cell engager) provided herein, any suitable concentration of nucleic acid can be used. For example, the pharmaceutical compositions provided herein may be formulated to be liquids containing about 0.5 mg to about 500 mg (e.g., about 1 mg to about 500 mg, about 10 mg to about 500 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 0.5 mg to about 250 mg, about 0.5 mg to about 150 mg, about 0.5 mg to about 100 mg, about 0.5 mg to about 50 mg, about 1 mg to about 300 mg, about 2 mg to about 200 mg, about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 150 mg, or about 150 mg to about 300 mg) of the nucleic acid provided herein per mL. In another example, the pharmaceutical compositions provided herein may be formulated to be solid or semi-solid containing about 0.5 mg to about 500 mg (e.g., about 1 mg to about 500 mg, about 10 mg to about 500 mg, about 50 mg to about 500 mg, about 100 mg to about 500 mg, about 0.5 mg to about 250 mg, about 0.5 mg to about 150 mg, about 0.5 mg to about 100 mg, about 0.5 mg to about 50 mg, about 1 mg to about 300 mg, about 10 mg to about 300 mg, about 25 mg to about 300 mg, about 50 mg to about 150 mg, or about 150 mg to about 300 mg).

[0151] In some cases, pharmaceutical compositions designed to include a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, cell engager, and / or ADC) provided herein can be formulated to include one or more agents that can reduce aggregation of the binding agent when formulated. Examples of such agents that can be used as described herein include, but are not limited to, methionine, arginine, lysine, aspartic acid, glycine, glutamic acid, and combinations thereof. In some cases, one or more of these amino acids can be included in the formulation at a concentration of about 0.5 mM to about 145 mM (e.g., about 1 mM to about 145 mM, about 10 mM to about 145 mM, about 100 mM to about 145 mM, about 0.5 mM to about 125 mM, about 0.5 mM to about 100 mM, about 0.5 mM to about 75 mM, or about 10 mM to about 100 mM).

[0152] The pharmaceutical compositions provided herein may be in any suitable form. For example, the pharmaceutical compositions provided herein may be designed to be liquid, semi-solid, or solid. In some cases, the pharmaceutical compositions provided herein may be a liquid solution (e.g., a solution for injection and / or infusion), a dispersion, a suspension, a tablet, a pill, a powder, a microemulsion, a liposome, or a suppository. In some cases, the pharmaceutical compositions provided herein may be lyophilized. In some cases, the pharmaceutical compositions provided herein (e.g., the pharmaceutical compositions provided herein comprising one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs)) may be formulated with a carrier or coating designed to protect against rapid release. For example, the pharmaceutical compositions provided herein may be formulated as controlled or regulated release formulations as described elsewhere (U.S. Patent Application Publication Nos. 2019 / 0241667, 2019 / 0233522, and 2019 / 0233498).

[0153] This document provides a method for the preparation of one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) (or a nucleic acid, vector, or host cell provided herein (e.g., CAR + Also provided are methods of administering a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, and / or host cell (e.g., CAR) provided herein) to a mammal (e.g., a human). + A composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, and / or host cell (e.g., a CAR)) can be administered to a mammal (e.g., a human) with cancer to treat the mammal. In some cases, one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, and / or host cell (e.g., a CAR)) can be administered to a mammal (e.g., a human) with cancer to treat the mammal. + A composition (e.g., a pharmaceutical composition provided herein) containing the cancer cells can be administered to a mammal (e.g., a human) to reduce the number of cancer cells in the mammal and / or to increase survival of the mammal afflicted with cancer.

[0154] One or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) (or a nucleic acid, vector, or host cell provided herein (e.g., a CAR +Any suitable cancer can be treated using a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein. For example, a composition (e.g., a pharmaceutical composition) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein can be administered to a mammal (e.g., a human) having cancer to treat the mammal. Examples of cancers that can be treated as described herein include, but are not limited to, lung cancer, prostate cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, vaginal cancer, cervical cancer, pancreatic cancer, head and neck cancer, blood cancer, skin cancer, brain cancer, bone cancer, and breast cancer. In some cases, a mammal (e.g., a human) having cancer (e.g., lung cancer, prostate cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, vaginal cancer, or cervical cancer) can be administered a composition (e.g., a pharmaceutical composition) containing one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) to treat the mammal (e.g., reduce the number of cancer cells in the mammal).

[0155] Any suitable viral infection can be ameliorated by the administration of one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) (or a nucleic acid, vector, or host cell provided herein (e.g., a CAR +For example, a mammal (e.g., a human) having an HIV viral infection, a hepatitis viral infection, a herpes viral infection, an influenza viral infection, a coronavirus infection, a cytomegalovirus infection, or a Hendra virus infection may be treated by administering to the mammal a composition (e.g., a pharmaceutical composition) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein.

[0156] Any suitable method can be used to administer a composition (e.g., a pharmaceutical composition) provided herein to a mammal (e.g., a human). For example, a composition provided herein (e.g., a pharmaceutical composition containing one or more binding agents provided herein, such as one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs provided herein) can be administered to a mammal (e.g., a human) intravenously (e.g., via intravenous injection or infusion), subcutaneously (e.g., via subcutaneous injection), intraperitoneally (e.g., via intraperitoneal injection), orally, via inhalation, or intramuscularly (e.g., via intramuscular injection). In some cases, the route and / or mode of administration of a composition (e.g., a pharmaceutical composition provided herein) can be tailored for the mammal being treated.

[0157] In some cases, one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) (or a nucleic acid, vector, or host cell provided herein (e.g., a CAR +An effective amount of a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR) provided herein) can be an amount that reduces the number of cancer cells and / or virus-infected cells in a mammal having cancer without causing significant toxicity to the mammal. In some cases, one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR) provided herein) can be an amount that reduces the number of cancer cells and / or virus-infected cells in a mammal having cancer without causing significant toxicity to the mammal. +An effective amount of a composition (e.g., a pharmaceutical composition provided herein) containing a cellular cellular antigen (cell) may be an amount that extends the survival of a mammal having cancer (and / or viral infection) compared to a control mammal having a similar cancer (and / or viral infection) and not treated with the composition. For example, an effective amount of a binding agent (e.g., an antibody, antigen-binding fragment, antibody domain, cell engager, and / or ADC) provided herein may be from about 0.001 mg / kg to about 100 mg / kg (e.g., from about 0.001 mg / kg to about 90 mg / kg, from about 0.001 mg / kg to about 80 mg / kg, from about 0.001 mg / kg to about 70 mg / kg, from about 0.001 mg / kg to about 60 mg / kg, from about 0.001 mg / kg to about 100 mg / kg (e.g., from about 0.001 mg / kg to about 150 mg / kg, from about 0.001 mg / kg to about 20 mg / kg, from about 0.001 mg / kg to about 30 mg / kg, from about 0.001 mg / kg to about 40 mg / kg, from about 0.001 mg / kg to about 50 mg / kg, from about 0.001 mg / kg to about 60 mg / kg, from about 0.001 mg / kg to about 70 mg / kg, from about 0.001 mg / kg to about 80 mg / kg, from about 0.001 mg / kg to about 90 mg / kg, from about 0.001 mg / kg to about 100 mg / kg). 001mg / kg ~ approx. 50mg / kg, approx. 0.001mg / kg ~ approx. 40mg / kg, approx. 0.001mg / kg ~ approx. 30mg / kg, approx. 0.005mg / kg ~ approx. 100mg / kg, approx. 0.01m g / kg~about 100mg / kg, about 0.05mg / kg~about 100mg / kg, about 0.1mg / kg~about 100mg / kg, about 0.5mg / kg~about 100mg / kg, about 1mg / kg~about 100m g / kg, about 5 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 25 mg / kg, about 0.1 mg / kg to about 30 mg / kg, about 0.15 mg / kg to about 25 mg / kg, about 0.2 mg / kg~about 20mg / kg, about 0.5mg / kg~about 20mg / kg, about 1mg / kg~about 30mg / kg, about 1mg / kg~about 25mg / kg, about 1mg / kg~about 20mg / kg, about 2mg / kg The effective amount may be about 20 mg / kg to about 20 mg / kg, about 5 mg / kg to about 30 mg / kg, about 10 mg / kg to about 30 mg / kg, about 15 mg / kg to about 30 mg / kg, about 20 mg / kg to about 30 mg / kg, about 3 mg / kg to about 30 mg / kg, about 0.5 mg / kg to about 10 mg / kg, about 1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 5 mg / kg, or about 1 mg / kg to about 3 mg / kg. The effective amount may remain constant or may be adjusted as a sliding scale or variable dose depending on the mammal's response to the treatment. The actual effective amount used for a particular application may be affected by a variety of factors.For example, the severity of the cancer when treating a mammal with cancer, the route of administration, the severity of the viral infection when treating a mammal with a viral infection, the route of administration, the age and general health of the mammal, the use of excipients, the possibility of co-administration with other therapeutic or prophylactic treatments, such as the use of other agents (e.g., checkpoint inhibitors), and the judgment of the treating physician may require an increase or decrease in the actual effective amount of the compositions provided herein to be administered (e.g., pharmaceutical compositions containing one or more binding agents provided herein).

[0158] In some cases, one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) (or a nucleic acid, vector, or host cell provided herein (e.g., a CAR + An effective frequency of administration of a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR) provided herein) can be a frequency that reduces the number of cancer cells (and / or virally infected cells) in a mammal having cancer (and / or viral infection) without causing significant toxicity to the mammal. In some cases, an effective frequency of administration of a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR) provided herein) can be a frequency that reduces the number of cancer cells (and / or virally infected cells) in a mammal having cancer (and / or viral infection) without causing significant toxicity to the mammal. +The effective frequency of administration of a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (cells) may be a frequency that extends the survival of a mammal having cancer (and / or viral infection) compared to a control mammal having a similar cancer (and / or viral infection) and not treated with the composition. For example, the effective frequency of administration of a pharmaceutical composition provided herein, e.g., a pharmaceutical composition containing one or more binding agents provided herein, may be from about twice daily to about once per year (e.g., from about twice daily to about once per month, from about twice daily to about once per week, from about once daily to about once per month, or from once daily to about once per week). In some cases, the frequency of administration of a pharmaceutical composition provided herein, e.g., a pharmaceutical composition containing one or more binding agents provided herein, may be daily. The frequency of administration of a pharmaceutical composition provided herein, e.g., a pharmaceutical composition containing one or more binding agents provided herein, may remain constant or may vary over the duration of the treatment. The actual effective frequency used for a particular application may be influenced by a variety of factors. For example, the severity of the cancer, the severity of the viral infection, the route of administration, the age and general health of the mammal, the use of excipients, the possibility of co-administration with other therapeutic or prophylactic treatments, such as the use of other agents (e.g., checkpoint inhibitors), and the judgment of the treating physician may require an increase or decrease in the actual effective frequency of administration of the compositions provided herein (e.g., pharmaceutical compositions containing one or more binding agents provided herein).

[0159] In some cases, one or more binding agents provided herein (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) (or a nucleic acid, vector, or host cell provided herein (e.g., a CAR +The effective duration of administration of a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR)) can be a duration that reduces the number of cancer cells (and / or virus-infected cells) in the mammal without causing significant toxicity to the mammal. In some cases, the effective duration of administration of a composition (e.g., a pharmaceutical composition provided herein) containing one or more binding agents (e.g., one or more antibodies, one or more antigen-binding fragments, one or more antibody domains, one or more cell engagers, and / or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR)) can be a duration that reduces the number of cancer cells (and / or virus-infected cells) in the mammal without causing significant toxicity to the mammal. + The effective duration of administration of a composition (e.g., a pharmaceutical composition provided herein) containing a binding agent (cell) may be a duration that extends the survival of a mammal having cancer (and / or viral infection) compared to a control mammal having a similar degree of cancer (and / or viral infection) and not treated with the composition. For example, the effective duration of administration of a pharmaceutical composition provided herein, e.g., a pharmaceutical composition containing one or more binding agents provided herein, may vary from a single time point administration to several weeks to several months (e.g., 4-12 weeks). The actual effective duration used for a particular application may be influenced by a number of factors. For example, the severity of the cancer, the severity of the viral infection, the route of administration, the age and general health of the mammal, the use of excipients, the possibility of co-administration with other therapeutic or prophylactic treatments, e.g., the use of other agents (e.g., checkpoint inhibitors), and the judgment of the treating physician may require an extension or shortening of the actual effective duration of administration of a composition provided herein (e.g., a pharmaceutical composition containing one or more binding agents provided herein).

[0160] In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) can be used to detect the presence or absence of a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) in vitro, in situ, or in vivo (e.g., for in vivo imaging in a mammal, e.g., a human). For example, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) can be designed to include a label (e.g., a covalently attached radioactive, enzymatic, chromogenic, or fluorescent label). The labeled binding agent can be used to detect the presence or absence of a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) in a biological sample in vitro. Examples of biological samples that can be evaluated using the binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) include, but are not limited to, serum samples, plasma samples, tissue samples, biopsy samples, cell line samples, and tissue culture samples. In some cases, the biological sample that can be evaluated as described herein can include mammalian body tissues and / or cells that can express a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide), such as leukocytes, T cells, NK cells, ovarian tissue or cells, prostate tissue or cells, cardiac tissue or cells, placental tissue or cells, pancreatic tissue or cells, liver tissue or cells, splenic tissue or cells, lung tissue or cells, breast tissue or cells, head and neck tissue or cells, endometrial tissue or cells, colonic tissue or cells, colorectal tissue or cells, cervical tissue or cells, gastric tissue or cells, or umbilical cord tissue or cells. In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) can be immobilized, for example, on a support, and retention of a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) from the biological sample on the support can be detected, and / or vice versa.In some cases, the binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) can be used in applications such as fluorescence polarization, microscopy, ELISA, centrifugation, chromatography, and / or cell sorting (e.g., fluorescence activated cell sorting).

[0161] In some cases, binding agents provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) containing a label (e.g., a covalently attached radioactive label) can be used to detect the presence or absence of a CD94 / NKG2A polypeptide (e.g., a human CD94 / NKG2A polypeptide) in a mammal (e.g., a human). For example, a labeled (e.g., covalently labeled) binding agent provided herein (e.g., antibodies, antigen-binding fragments, and / or antibody domains) having a radiolabel or an MRI-detectable label can be administered to a mammal (e.g., a human), and the mammal can be evaluated using a means for detecting the detectable label. In some cases, the mammal can be scanned to evaluate the location of the labeled binding agent provided herein in the mammal. For example, the mammal can be imaged using NMR or other tomography techniques.

[0162] Examples of labels that can be attached (e.g., covalently or non-covalently attached) to the binding agents (e.g., antibodies, antigen-binding fragments, and / or antibody domains) provided herein include, but are not limited to, radiolabels, e.g. 131 I, 111 In, 123 I, 99m Tc, 32 P, 33 P, 125 I, 3 H, 14 C, and 188These include Rh, fluorescent labels such as fluorescein and rhodamine, nuclear magnetic resonance active labels, positron emitting isotopes detectable by a positron emission tomography ("PET") scanner, chemiluminescent markers such as luciferin, and enzymatic markers such as peroxidase or phosphatase. In some cases, short-range luminescent markers, such as isotopes detectable by short-range detector probes, can be used.

[0163] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0164] [Example] [Example 1] Obtaining a binding agent capable of binding to human CD94 / NKG2A polypeptide A vector was generated to express a CD94 / NKG2A polypeptide with amino acid residues 57-179 of CD94 and 113-233 of NKG2A fused to human IgG1 Fc. A GSGGSGG (SEQ ID NO: 258) linker was used between CD94 and NKG2A (Figure 1). Using this CD94 / NKG2A-IgG1 fusion polypeptide, one Fab antibody fragment (clone #1, Figures 2A-B) was identified. To identify this Fab, the CD94 / NKG2A-IgG1 fusion polypeptide was subjected to a three-round panning process.

[0165] The phage library was preblocked with 3% (w / v) bovine serum albumin (BSA) in PBS for 1 h at 25° C. Blocked phages were incubated with 10 nM biotinylated single-chain CD94 / NKG2A-Fc in the presence of 50 nM single-chain CD94 / NKG2C-Fc for 1 h at 25° C. Bound phages were separated by streptavidin-coated magnetic beads and washed 10 times with 1 mL PBS pH 7.4 containing 0.1% Tween-20 (w / v). Elution of bound phages was performed by adding 1 μM anti-NKG2A antibody or 5 nM HLA-E (HLA-E*01:01 HLA-A leader 3-11 monomer-VMAPRTLVL) tetramer. For the second and third rounds of panning, reduced concentrations of biotinylated single-chain CD94 / NKG2A (5 nM and 1 nM, respectively) and increased competitor ratios of single-chain CD94 / NKG2C were applied. After three rounds of panning, individual clones were analyzed by ELISA and selected clones were then sequenced after plasmid rescue.

[0166] Fab clone #1 bound to the CD94 / NKG2A polypeptide with high affinity and specificity. Fab clone #1 had an EC of 2.7 nM in ELISA against CD94 / NKG2A. 50 IgG clone #1 showed a K value of 1.1 nM in a BLItz Biolayer Interferometer, but clone #1 did not cross-react with CD94 / NKG2C polypeptide in an ELISA experiment (Figure 23). DThe equilibrium dissociation constants (equilibrium dissociation constants) were shown (Figure 25). Furthermore, IgG clone #1 competed with both tetrameric HLA-E for binding to the CD94 / NKG2A polypeptide, but did not affect binding to CD94 / NKG2C (Figure 26). These results demonstrate that clone #1 is specific for NKG2A and not NKG2C, and that it binds to the HLA-E binding interface of NKG2A, resulting in blocking by clone #1 of the inhibitory interaction between NKG2A and HLA-E (or possibly other HLA molecules or receptors that bind to a similar interface of NKG2A instead of HLA-E).

[0167] To improve the binding affinity of clone #1, yeast display technology and a light chain shuffling strategy were applied to generate clone #2. IgG clone #2 showed improved binding compared to clone #1 by ELISA and had an EC 50 was measured to be 0.6 nM (Figure 27). Clone #2 also showed no cross-reactivity to NKG2C. These results demonstrate that clone #2 retains specificity for NKG2A. IgG clone #2 also showed improved competition for binding to CD94 / NKG2A polypeptide compared to the original clone #1, which was comparable to that of monalizumab (Figure 28).

[0168] To confirm the binding of the antibodies (monalizumab analogues, clone #1 and clone #2) to cell surface NKG2A, primary NK cells were enriched from normal human peripheral blood mononuclear cells (PBMCs) using an NK cell isolation kit. Primary NK cells were treated with the antibodies (10 nM) for 1 hour at 4° C. and then stained with Alexa647-conjugated goat anti-human IgG for 0.5 hours at 4° C. Staining was detected for monalizumab analogues, clone #1 and clone #2, which indicated that all three antibodies bound to the intact NKG2A receptor on the NK cell surface as well as recombinant NKG2A (FIG. 24, FIG. 29 and FIG. 30).

[0169] [Example 2] Functional assays with binding agents capable of binding to human CD94 / NKG2A polypeptides To perform functional assays for clones #1 and #2, purified human primary NK cells from PBMC were used as effector cells. NK cells were activated with 50 IU / mL hIL-2 and treated with antibodies (monalizumab analogs, clone #1, and clone #2) in the absence or presence of HLA-E positive H2030 cells for 24 hours at 37°C (E:T ratio 5:1). First, changes in the NKG2A expression level of NK cells were examined. Antibody-treated NK cells were washed with cold PBS, and then NKG2A on the cell surface was detected using FITC- or PE-conjugated anti-human CD159a (NKG2A) antibody. For gating of NK cells in the co-culture system with H2030 cells, APC-conjugated anti-human CD45 antibody was used. Antibody treatment induced downregulation of NKG2A on the cell surface, which was probably caused by receptor internalization (Figure 31). Next, to examine the activation of NK cells, IFNγ and granzyme B were analyzed by intracellular staining. GolgiPlug containing Brefeldin A was added during the last 4 hours of culture to inhibit protein transport from the endoplasmic reticulum to the Golgi apparatus. All intracellular staining was performed using the BD cytofix / cytoperm kit. After antibody treatment, cells were fixed and stained intracellularly with PE-conjugated anti-human IFNγ and PE-conjugated anti-human granzyme B antibodies. Throughout all flow cytometry experiments, labeled cells were washed twice and analyzed on a BD LSR II flow cytometer. The observed flow cytometry results demonstrated that treatment with clone #2 or monalizumab analogs activated NK cells and increased the secretion of IFNγ and granzyme B with or without co-culture with H2030 cell line, while treatment with clone #1 and durvalumab (anti-PD-L1 antibody) showed no activation of NK cells (Figure 32). Clone #1 did not show statistical significance, but a trend towards activation was observed compared to durvalumab.These results also demonstrate that targeting NKG2A activates NK cells by blocking the interaction with HLA-E and / or decreasing the surface expression level of NKG2A, and that binding affinity is involved in NK cell activation.

[0170] [Example 3] Design of cell engagers from binders capable of binding to human CD94 / NKG2A polypeptide To demonstrate the design of a bispecific NK cell / T cell engager (BiKE / BiTE, also referred to herein as BINK) capable of binding to CD94 / NKG2A polypeptide on NK cells or T cells and tumor-associated antigens (TAA) on cancer cells, the following was performed. CEACAM5 (CD66e) polypeptide was selected as an example of TAA. Anti-CEACAM5 scFv was fused to the N-terminus of the light chain of clone #1 or clone #2 via a GSGGGS (SEQ ID NO: 318) linker (see, for example, Figures 17A and 17B). Plasmid DNA encoding cloned BINK was transfected into HEK239F cells and expressed for 5-7 days after transfection. Expressed BINK polypeptide was purified by affinity chromatography with protein A resin. Elution of bound BINK polypeptide was performed by adding 50 mM glycine buffer pH 3.0, followed by changing the storage buffer to phosphobuffered saline pH 7.4 (PBS) by a PD-10 desalting column. Protein purity was estimated by SDS-PAGE or size-exclusion chromatography loaded with Superdex 200 increase 10 / 300 GL. The concentration of each BINK polypeptide was determined by Nano Drop Spectrophotometer 2000C. The binding and specificity of each BINK polypeptide to single-chain CD94 / NKG2A polypeptide, single-chain CD94 / NKG2C polypeptide, CEACAM5 A3B3 domain, or CEACAM6 AB domain was analyzed by indirect ELISA. Briefly, Fc fusion proteins were coated at 200 ng / well (volume 50 μL) in PBS onto 96-well plates at 25°C for 2 hours. Blocking was performed with 3% skim milk in PBS at 4°C overnight. The next day, various concentrations of BINK polypeptide were added to the antigen-coated plates and incubated at 25°C for 1 hour. After washing three times with PBS-T (PBS pH 7.4 containing 0.1% Tween-20), HRP-conjugated goat anti-human kappa antibody (1:3000 dilution) was added to detect binding of BINK polypeptide and incubated for 1 h at 25°C.After washing three times with PBS-T, 50 μL of TMB (Thermo, PI34028) was added as a substrate, and the enzyme reaction was stopped by adding 2 N sulfonic acid. The BINK molecule showed binding specificity to NKG2A and CECAM5-A3B3 as designed (Figure 33).

[0171] [Example 4] Design of further cell engagers from binders capable of binding to human CD94 / NKG2A polypeptides To demonstrate the design of further bispecific NK cell / T cell engagers (BiKE / BiTE, also referred to herein as BiNK) capable of binding to CD94 / NKG2A polypeptide on NK cells or T cells and tumor-associated antigens (TAA) on cancer cells, the following was performed. In one example, HER2 polypeptide was selected as an example of TAA. In another example, EGFR polypeptide was selected as an example of TAA.

[0172] Anti-HER2 scFv was fused to the N-terminus of the light chain of clone #1 via a GSGGGS (SEQ ID NO: 318) linker to create BiNK designated "BiNK (anti-HER2 x anti-NKG2A)" (see, e.g., Figures 17A and 17C). Anti-EGFR scFv was fused to the N-terminus of the light chain of clone #1 via a GSGGGS (SEQ ID NO: 318) linker to create BiNK designated "BiNK (anti-EGFR x anti-NKG2A)" (see, e.g., Figures 17A and 17D). Briefly, plasmid DNA encoding the cloned BiNK was transfected into HEK239F cells and expressed for 5 to 7 days after transfection. The expressed BiNK polypeptides were purified and the binding of each BiNK polypeptide to single-chain CD94 / NKG2A polypeptide, single-chain CD94 / NKG2C polypeptide, HER2 polypeptide, or EGFR polypeptide was analyzed by indirect ELISA.

[0173] BiNK (anti-HER2 x anti-NKG2A) bound to single-chain CD94 / NKG2A polypeptides and HER2 (Figures 34A and 34B), but not to single-chain CD94 / NKG2C polypeptides. Flow cytometry also confirmed that BiNK (anti-EGFR x anti-NKG2A) bound to single-chain CD94 / NKG2A polypeptides and EGFR, but not to single-chain CD94 / NKG2C polypeptides.

[0174] Primary NK cells express CD94 / NKG2A polypeptides, and both BiNK (anti-HER2 x anti-NKG2A) and BiNK (anti-EGFR x anti-NKG2A) as well as clone #2 in IgG format showed binding to primary NK cells (Figure 35A). A549 cells express both HER2 and EGFR polypeptides, and both BiNK (anti-HER2 x anti-NKG2A) and BiNK (anti-EGFR x anti-NKG2A) as well as cetuximab showed binding to A549 cells (Figure 35B). Farage cells do not express HER2, EGFR, or NKG2A polypeptides, and neither BiNK (anti-HER2 x anti-NKG2A), BiNK (anti-EGFR x anti-NKG2A), nor cetuximab showed any appreciable binding to Farage cells (Figure 35B).

[0175] When cell killing by NK cells in the presence of various binders was assessed, BiNK (anti-HER2 x anti-NKG2A), BiNK (anti-EGFR x anti-NKG2A), cetuximab, and pertuzumab promoted killing of A549 cells (Figure 36).

[0176] In co-incubation cultures of primary NK cells and A549 cells, BiNK (anti-EGFR × anti-NKG2A) treatment significantly increased CD56 + CD16A on NK cells + Increase NK cell population (%) and CD56 + NKG2A in NK cells + BiNK reduced the CD16 cell population (%) but did not alter the NKG2C population (Figure 37). +When cell killing was assessed in different timed treatment schedules by population expansion (Figure 38A), BiNK (anti-HER2 x anti-NKG2A) enhanced the ADCC activity of anti-HER2 IgG1 pertuzumab (Figure 38B).

[0177] To evaluate how much NKG2A binding capacity is required for NK cell activation, NKG2A binders with various NKG2A binding capacities were constructed. Both Clone#2 IgG (anti-NKG2A) and BiNK (anti-EGFR x anti-NKG2A), which have bivalent binding to NKG2A, downregulated the expression of NKG2A (Figure 39) and increased the production of IFNγ, TNFα, and granzyme B (GrzB) by primary NK cells (Figure 40). Clone#2-SA bead mix, which has multivalent binding mimicking HLA-E / NKG2A interaction, also reduced the expression level of NKG2A (Figure 39), yet reduced TNFα and GrzB (Figure 40). These results demonstrate that bivalent binding to NKG2A contributes to NK cell activation.

[0178] [Example 5] Methods and Materials Purification of Fab and IgG1-LALA-PG (L234A, L235A, and P329G mutations) The plasmids were transformed into HB2151 E. coli competent cells, and colonies were then selected on ampicillin-containing LB plates (final concentration 100 μg / mL) overnight in a 37° C. incubator. The next day, colonies were inoculated into liquid LB+ampicillin medium and cultured in a shaking incubator at 37° C. Approximately 4×10 8 An OD of approximately 0.4-0.6 corresponds to cells / mL 600Isopropyl β-D-1-thiogalactopyranoside (IPTG) was added to the culture at a final concentration of 0.1 mM at 200 °C. The culture was transferred to a shaking incubator set at 30 °C and 200 rpm and incubated overnight. The next day, E. coli cells were harvested and resuspended in 1 / 10 volume of periplasmic extraction buffer containing polymyxin B (0.5 mg / mL in PBS, pH 7.4) and then incubated on ice for 1 h. The supernatant was collected by centrifugation at 12,000 × g for 10 min and then loaded onto pre-packed Ni-NTA resin. Bound Fab was eluted by adding 300 mM imidazole in PBS (pH 7.4), and then imidazole was removed using a PD-10 desalting column (GE, 45-000-148).

[0179] For preparation of IgG, IgG cloned plasmid DNA was transfected into HEK239F cells and expressed for 5–7 days after transfection. Expressed IgG was purified by affinity chromatography with Protein A resin (Captiva, NC0997253). Elution of bound IgG was performed by adding 50 mM glycine buffer pH 3.0, and then the storage buffer was changed to phosphobuffered saline pH 7.4 (PBS) by PD-10 desalting column. Purity of protein was estimated by SDS-PAGE or size-exclusion chromatography packed with Superdex 200 increase 10 / 300 GL (GE healthcare, 28990944). Concentration of each protein was determined by Nano Drop spectrophotometer 2000C (Thermo, ND2000C).

[0180] Binding of Fab or IgG in Enzyme-Linked Immunosorbent Assay (ELISA) The binding and specificity of Fab or IgG to single-chain CD94 / NKG2A-Fc or single-chain CD94 / NKG2C-Fc was analyzed by indirect ELISA. Briefly, Fc fusion proteins were coated at 200 ng / well (volume 50 μL) in PBS onto 96-well plates (Corning, 3690) for 2 h at room temperature. Blocking was performed with 3% BSA in PBS overnight at 4°C. The next day, various concentrations of Fab or IgG were applied to the antigen-coated plates and incubated for 1 h at 25°C. After three washes, anti-FLAG mouse antibody (M2 clone)-HRP conjugate (Sigma, A8592, 1:3000 dilution) or anti-human kappa goat antibody-HRP conjugate (Invitrogen, A18853, 1:3000 dilution) was applied to detect the binding of Fab or IgG. The same volume of TMB (Thermo, PI34028) was added as substrate, and then the enzymatic reaction was stopped by adding 2N sulfonic acid.

[0181] cell line H2030, A549, Farage, and 293T cells were purchased from ATCC. H2030, A549, and Farage cells were cultured in RPMI 1640 (ATCC) supplemented with 10% v / v FBS (Gibco) and 1% penicillin-streptomycin (P / S, Gibco). 293T cells were maintained in DMEM (Gibco) supplemented with 10% v / v FBS and 1% P / S. Primary NK cells were cultured in MACS basal NK medium with supplementation (Miltenyi Biotec, 130-114-429), 10% v / v human serum (Sigma-Aldrich), and hIL-2 (50 IU / mL, Miltenyi Biotec).

[0182] Flow cytometry analysis Primary NK cells were enriched from normal human peripheral blood mononuclear cells (PBMCs, Zenbio Inc.) using an NK cell isolation kit (Miltenyi Biotec, 130-092-657). The purity of the isolated NK cells was confirmed by staining with APC-conjugated anti-human CD56 and PE-conjugated anti-human CD16 antibodies. NKG2A of primary NK cells + To determine populations, cells were stained with Alexa488 (AR488)-conjugated anti-NKG2A IgG1. To confirm cell surface binding of antibodies (e.g., monalizumab, clone #1 IgG, and clone #2 IgG), cells were treated with antibodies (10 nM) for 1 h at 4°C and then stained with Alexa647-conjugated goat anti-human IgG (Invitrogen, A21445) for 0.5 h at 4°C. To examine NKG2A expression levels on NK cells, enriched NK cells were activated with 50 IU / mL hIL-2 and treated with the indicated antibodies (100 nM) for 24 h at 37°C. After incubation with antibodies, cells were washed with cold PBS and then cell surface NKG2A was detected using FITC- or PE-conjugated anti-human CD159a (NKG2A) antibodies. For gating of NK cells in co-cultures with H2030 or A549 cells, an APC-conjugated anti-human CD45 antibody was used. Monoclonal antibodies specific for CD56 (12-0567-42), CD16 (12-0168-42), CD159a (130-113-566), and CD45 (17-0459-42) were purchased from Thermofisher. For intracellular staining, GolgiPlug containing Brefeldin A (Invitrogen, 00-4506-51) was added during the last 4 hours of culture to inhibit protein transport from the endoplasmic reticulum to the Golgi apparatus.

[0183] All intracellular staining was performed using the BD cytofix / cytoperm kit (BD Biosciences, BD554714). Monoclonal antibodies specific for IFNγ (12-7319-42) and granzyme B (12-8896-42) were purchased from Thermofisher. The anti-PD-L1 antibody durvalumab was purchased from Selleckchem (A2013). Data were acquired using a flow cytometer BD LSR II (San Jose, CA) and analyzed by FlowJo 10.7.1 (Tree Star).

[0184] Functional assays in peripheral human NK cells Purified human NK cells from PBMC were used as effector cells. NK cells were incubated with target cell lines (H2030 or A549) in the presence of 50 IU / mL hIL-2 for 24 hours at 37°C (E:T ratio 5:1 or 2:1). Cells were distributed in 96-well plates with or without 100 nM of the indicated binders (e.g., monalizumab, clone #1 IgG, clone #2 IgG, durvalumab, or BiNK) for 24 hours at 37°C. After treatment with binders, cells were fixed and stained intracellularly with PE-conjugated anti-human IFNγ, PE-conjugated anti-human TNFα, and PE-conjugated anti-human granzyme B antibodies. Cells were washed twice and analyzed by BD LSR II flow cytometry.

[0185] In vitro cytotoxicity assay The ADCC or cytotoxicity of effector cells incubated with anti-NKG2A binders carrying the LALA-PG mutation was measured by the release of cytoplasmic LDH from killed target cells using the LDH-Glo Cytotoxicity Assay Kit (Promega, J2381). Purified NK cells (Zenbio Inc.) from normal PBMCs were used as effector cells, and A549 or 293T cells (1 × 10 in 96-well plates) as target cells in the presence of the indicated binders were incubated with NK cells (1 × 10 in 96-well plates) as effector cells. 4Cells / well) at an effector to target (E:T) ratio of 5:1 or 2:1 for 4 hours at 37° C. Percent cytotoxicity was calculated by the following formula: 100×(experimental effector min−target min) / (target max−target min).

[0186] Other embodiments While the present invention has been described in conjunction with its detailed description, it should be understood that the foregoing description is for illustrative purposes only and is not intended to limit the scope of the invention as defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with the addition, deletion or substitution of one, two or three amino acids), SEQ ID NO:2 (or SEQ ID NO:2 with the addition, deletion or substitution of one, two or three amino acids) and SEQ ID NO:3 (or SEQ ID NO:3 with the addition, deletion or substitution of one, two or three amino acids), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with the addition, deletion or substitution of one, two or three amino acids), SEQ ID NO:10 (or SEQ ID NO:10 with the addition, deletion or substitution of one, two or three amino acids) and SEQ ID NO:11 (or SEQ ID NO:11 with the addition, deletion or substitution of one, two or three amino acids); or (ii) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 (or SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). An antibody comprising:

2. 2. The antibody of claim 1, having the ability to bind to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, or SEQ ID NO:

259.

3. An antibody according to any one of claims 1 to 2, comprising the heavy chain variable domain or region of (i).

4. The antibody of claim 3, wherein the heavy chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

8.

5. An antibody described in any one of claims 1 to 2, comprising the light chain variable domain or region of (i).

6. The antibody of claim 5, wherein the light chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

16.

7. An antibody described in any one of claims 1 to 2, comprising the heavy chain variable domain or region of (ii).

8. The antibody of claim 7, wherein the heavy chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

24.

9. An antibody described in any one of claims 1 to 2, comprising the light chain variable domain or region of (ii).

10. The antibody of claim 9, wherein the light chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

32.

11. (i) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:1 (or SEQ ID NO:1 with the addition, deletion or substitution of one, two or three amino acids), SEQ ID NO:2 (or SEQ ID NO:2 with the addition, deletion or substitution of one, two or three amino acids) and SEQ ID NO:3 (or SEQ ID NO:3 with the addition, deletion or substitution of one, two or three amino acids), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO:9 (or SEQ ID NO:9 with the addition, deletion or substitution of one, two or three amino acids), SEQ ID NO:10 (or SEQ ID NO:10 with the addition, deletion or substitution of one, two or three amino acids) and SEQ ID NO:11 (or SEQ ID NO:11 with the addition, deletion or substitution of one, two or three amino acids); or (ii) a heavy chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 17 (or SEQ ID NO: 17 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 18 (or SEQ ID NO: 18 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 19 (or SEQ ID NO: 19 with one, two or three amino acid additions, deletions or substitutions), and a light chain variable domain or region comprising the amino acid sequence set forth in SEQ ID NO: 25 (or SEQ ID NO: 25 with one, two or three amino acid additions, deletions or substitutions), SEQ ID NO: 26 (or SEQ ID NO: 26 with one, two or three amino acid additions, deletions or substitutions) and SEQ ID NO: 27 (or SEQ ID NO: 27 with one, two or three amino acid additions, deletions or substitutions). An antigen-binding fragment comprising:

12. 12. The antigen-binding fragment of claim 11, having the ability to bind to SEQ ID NO: 74 and SEQ ID NO: 75, SEQ ID NO: 256 and SEQ ID NO: 257, or SEQ ID NO:

259.

13. An antigen-binding fragment according to any one of claims 11 to 12, comprising the heavy chain variable domain or region of (i).

14. The antigen-binding fragment of claim 13, wherein the heavy chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

8.

15. An antigen-binding fragment according to any one of claims 11 to 12, comprising the light chain variable domain or region of (i).

16. The antigen-binding fragment of claim 15, wherein the light chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

16.

17. An antigen-binding fragment according to any one of claims 11 to 12, comprising the heavy chain variable domain or region of (ii).

18. 18. The antigen-binding fragment of claim 17, wherein the heavy chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

24.

19. An antigen-binding fragment according to any one of claims 11 to 12, comprising the light chain variable domain or region of (ii).

20. 20. The antigen-binding fragment of claim 19, wherein the light chain variable domain or region comprises an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:

32.

21. The antibody according to any one of claims 1 to 10, which is a monoclonal antibody.

22. 22. An antibody according to any one of claims 1 to 10 and 21, which is an scFv antibody.

23. 21. An antigen-binding fragment according to any one of claims 11 to 20, which is monoclonal.

24. 24. An antigen-binding fragment according to any one of claims 11 to 20 and 23, which is a Fab.

25. 25. A chimeric antigen receptor comprising an antigen-binding domain, a hinge, a transmembrane domain and one or more signaling domains, wherein the antigen-binding domain comprises an antibody or antigen-binding fragment of any one of claims 1 to 24.

26. The chimeric antigen receptor of claim 25, wherein the antigen-binding domain comprises an scFv having the ability to bind to a CD94 / NKG2A polypeptide.

27. The chimeric antigen receptor of any one of claims 25 to 26, wherein the hinge comprises the hinge depicted in Figure 13.

28. 28. The chimeric antigen receptor of any one of claims 25 to 27, wherein the transmembrane domain comprises the transmembrane domain depicted in Figure 14.

29. 29. The chimeric antigen receptor of any one of claims 25 to 28, comprising one or more signaling domains as described in Figure 15.

30. A cell comprising a chimeric antigen receptor according to any one of claims 25 to 29.

31. 31. The cell of claim 30, which is a T cell, a stem cell or a NK cell.

32. 25. A cell engager comprising a first antigen-binding domain, a linker and a second antigen-binding domain, wherein the first antigen-binding domain comprises the antibody or antigen-binding fragment of any one of claims 1 to 24.

33. The cell engager of claim 32, wherein the first antigen-binding domain comprises an scFv capable of binding to a CD94 / NKG2A polypeptide.

34. 33. The cell engager of claim 32, wherein the first antigen-binding domain is an IgG having the ability of binding to a CD94 / NKG2A polypeptide.

35. The cell engager of any one of claims 32 to 34, wherein the linker comprises the linker of Figure 10 or Figure 13.

36. The cell engager of any one of claims 32 to 35, wherein the second antigen-binding domain binds to an antigen of interest.

37. 37. The cell engager of claim 36, wherein the antigen of interest is expressed on the surface of a cancer cell or a virus-infected cell.

38. 37. The cell engager of claim 36, wherein the antigen of interest is an antigen of interest selected from the group consisting of an EGFR polypeptide, a HER2 polypeptide, a CEACAM5 polypeptide, a CEACAM7 polypeptide, a CD19 polypeptide, a CD22 polypeptide, a CD274 polypeptide, a CD276 polypeptide, a PSMA polypeptide, a PSCA polypeptide, an ADAM10 polypeptide, a methotelin polypeptide, a GPC2 polypeptide, a FGFR polypeptide, a VEGFR polypeptide, an IGFR polypeptide, an HIV gp120 polypeptide, an HIV gp160 polypeptide, and a SARS-CoV-2 RBD polypeptide.

39. The cell engager of any one of claims 36 to 38, wherein the second antigen-binding domain is as shown in Figure 20.

40. 40. The cell engager of claim 39, wherein the second antigen binding domain comprises SEQ ID NOs: 263 and 264.

41. 40. The cell engager of claim 39, wherein the second antigen binding domain comprises SEQ ID NOs: 265 and 266.

42. 42. The cell engager of any one of claims 32 to 41, comprising a third antigen-binding domain.

43. The cell engager of claim 42, wherein the third antigen-binding domain binds to a polypeptide expressed on the surface of a NK cell.

44. The cell engager of claim 43, wherein the polypeptide expressed on the surface of a NK cell is a CD16a, NKG2A, NKG2D, NKp30, NKp44 or NKp46 polypeptide.

45. 44. The cell engager of claim 43, wherein the third antigen binding domain is the antigen binding domain depicted in Figure 19.

46. A nucleic acid comprising a nucleic acid sequence encoding at least a portion of the antibody or antigen-binding fragment of any one of claims 1 to 24.

47. 47. The nucleic acid of claim 46, wherein the nucleic acid sequence encodes the heavy chain variable domain or region of any one of (i) to (ii) of claim 1.

48. 48. The nucleic acid of any one of claims 46 to 47, wherein said nucleic acid sequence encodes said light chain variable domain or region of any one of (i) to (ii) of claim 1.

49. The nucleic acid of any one of claims 46 to 48, which is a viral vector.

50. 49. The nucleic acid of any one of claims 46 to 48, which is a phagemid.

51. 46. ​​A nucleic acid comprising a nucleic acid sequence encoding the chimeric antigen receptor of any one of claims 25 to 29 or the cell engager of any one of claims 32 to 45.

52. 52. The nucleic acid of claim 51, which is a viral vector.

53. 52. The nucleic acid of claim 51, which is a phagemid.

54. A host cell comprising the nucleic acid of any one of claims 46 to 52.

55. A host cell expressing a chimeric antigen receptor of any one of claims 25 to 29 or a cell engager of any one of claims 32 to 45.

56. 56. The host cell according to any one of claims 54 to 55, which is a T cell, a stem cell or a NK cell.

57. 25. An antibody drug conjugate (ADC) comprising an antigen-binding domain covalently linked to a drug, wherein the antigen-binding domain comprises the antibody or antigen-binding fragment of any one of claims 1 to 24.

58. 58. The ADC of claim 57, wherein the antigen-binding domain comprises an scFv having the ability of binding to a CD94 / NKG2A polypeptide.

59. 58. The ADC of claim 57, wherein the antigen-binding domain is an IgG having the ability of binding to a CD94 / NKG2A polypeptide.

60. 60. The ADC of any one of claims 57 to 59, wherein the drug is selected from the group consisting of BMS1166, BMS202, IL-2, and IL-12.

61. A composition comprising an antibody or antigen-binding fragment according to any one of claims 1 to 24.

62. 62. The composition of claim 61, comprising an antibody according to any one of claims 1 to 10, 21 and 22.

63. 62. The composition of claim 61, comprising an antigen-binding fragment of any one of claims 11 to 20, 23 and 24.

64. 46. ​​A composition comprising a cell engager according to any one of claims 32 to 45.

65. A composition comprising a cell according to any one of claims 30, 31 and 53 to 56.

66. A composition comprising the ADC of any one of claims 57-60.

67. 67. The composition of any one of claims 61 to 66, comprising a checkpoint inhibitor.

68. 68. The composition of claim 67, wherein the checkpoint inhibitor is selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189, and ipilimumab.

69. 70. A method of treating a mammal having cancer or a viral infection comprising administering to said mammal a composition of any one of claims 61-68.

70. 70. The method of claim 69, wherein the mammal is a human.

71. 71. The method of any one of claims 69 to 70, wherein the method comprises treating the mammal with cancer.

72. 72. The method of claim 71, wherein the cancer is selected from the group consisting of lung cancer, prostate cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, vaginal cancer and cervical cancer.

73. 73. The method of any one of claims 69-72, wherein the number of cancer cells or virally infected cells in said mammal is reduced following said administering step.

74. 1. A method of treating a mammal having cancer or a viral infection, comprising: (a) administering to said mammal a composition according to any one of claims 98 to 104; and (b) administering to the mammal a composition comprising a checkpoint inhibitor. The method includes:

75. 75. The method of claim 74, wherein the mammal is a human.

76. 76. The method of any one of claims 74 to 75, wherein the method comprises treating the mammal with cancer.

77. 77. The method of claim 76, wherein the cancer is selected from the group consisting of lung cancer, prostate cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, vaginal cancer and cervical cancer.

78. 78. The method of any one of claims 74 to 77, wherein the checkpoint inhibitor is selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostallimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189 and ipilimumab.

79. 79. The method of any one of claims 74-78, wherein the number of cancer cells or virus-infected cells in the mammal is reduced following said administering steps (a) and (b).

80. 25. A method for binding a binding molecule to a CD94 / NKG2A polypeptide, comprising the step of contacting said CD94 / NKG2A polypeptide with an antibody or antigen-binding fragment of any one of claims 1 to 24.

81. 81. The method of claim 80, wherein said contacting step is performed in vitro.

82. 81. The method of claim 80, wherein said contacting step is performed in vivo.

83. 83. The method of claim 82, wherein the contacting step is performed in a mammal by administering the antibody or antigen-binding fragment to the mammal.

84. 85. The method of claim 84, wherein the mammal is a human.

85. 61. A method of binding a binding molecule to a CD94 / NKG2A polypeptide, comprising the step of contacting said CD94 / NKG2A polypeptide with a chimeric antigen receptor of any one of claims 25-29, a cell engager of any one of claims 32-45, or an ADC of any one of claims 57-60.

86. 86. The method of claim 85, wherein said contacting step is performed in vitro.

87. 86. The method of claim 85, wherein said contacting step is performed in vivo.

88. 88. The method of claim 87, wherein the contacting step is performed in a mammal by administering to the mammal the chimeric antigen receptor, the cell engager or the ADC.

89. 89. The method of claim 88, wherein the mammal is a human.

90. A composition comprising an antibody or antibody-binding fragment, wherein the antibody or antibody-binding fragment has the ability to bind to a CD94 / NKG2A polypeptide present on an NK cell or a T cell without inhibiting the cytotoxic activity of the NK cell or the T cell.

91. The composition of claim 90, wherein the CD94 / NKG2A polypeptide is a human CD94 / NKG2A polypeptide.

92. The composition of any one of claims 90 to 91, wherein the antibody or antibody-binding fragment binds to the NK cell.

93. The composition of any one of claims 90 to 92, wherein the antibody or antibody binding fragment binds to the T cell.

94. The composition of any one of claims 90 to 93, wherein the antibody is an antibody of any one of claims 1 to 10, 21 and 22, and the antigen-binding fragment is an antigen-binding fragment of any one of claims 11 to 20, 23 and 24.

95. A composition comprising an antibody or antibody-binding fragment, wherein the antibody or antibody-binding fragment has the ability to bind to a CD94 / NKG2A polypeptide present on an NK cell or a T cell and reduce the expression of the CD94 / NKG2A polypeptide on the surface of the NK cell or the T cell.

96. The composition of claim 95, wherein the CD94 / NKG2A polypeptide is a human CD94 / NKG2A polypeptide.

97. The composition of any one of claims 95 to 96, wherein the antibody or antibody-binding fragment binds to the NK cell.

98. The composition of any one of claims 95 to 97, wherein the antibody or antibody binding fragment binds to the T cell.

99. The composition of any one of claims 95 to 98, wherein the antibody is an antibody of any one of claims 1 to 10, 21 and 22, and the antigen-binding fragment is an antigen-binding fragment of any one of claims 11 to 20, 23 and 24.

100. A cell engager comprising a first antigen-binding domain, a linker, and a second antigen-binding domain, wherein the first antigen-binding domain is capable of binding to a CD94 / NKG2A polypeptide present on a NK cell or a T cell without inhibiting the cytotoxic activity of the NK cell or the T cell.

101. The cell engager of Claim 100, wherein said first antigen-binding domain comprises an scFv capable of binding to said CD94 / NKG2A polypeptide.

102. The cell engager of Claim 100, wherein said first antigen-binding domain comprises an IgG having the ability of binding to said CD94 / NKG2A polypeptide.

103. The cell engager of any one of claims 100-102, wherein the first antigen-binding domain comprises an antibody or antigen-binding fragment of any one of claims 1-24.

104. The cell engager of any one of claims 100-103, wherein the linker comprises a linker as depicted in Figure 10 or Figure 13.

105. The cell engager of any one of claims 100-104, wherein the second antigen-binding domain binds to an antigen of interest.

106. The cell engager of claim 105, wherein the antigen of interest is expressed on the surface of a cancer cell or a virus-infected cell.

107. The cell engager of claim 106, wherein the antigen of interest is an antigen of interest selected from the group consisting of an EGFR polypeptide, a HER2 polypeptide, a CEACAM5 polypeptide, a CEACAM7 polypeptide, a CD19 polypeptide, a CD22 polypeptide, a CD274 polypeptide, a CD276 polypeptide, a PSMA polypeptide, a PSCA polypeptide, an ADAM10 polypeptide, a mesothelin polypeptide, a GPC2 polypeptide, an FGFR polypeptide, a VEGER polypeptide, an IGFR polypeptide, an HIV gp120 polypeptide, an HIV gp160 polypeptide, and a SARS-CoV-2 RBD polypeptide.

108. The cell engager of any one of claims 100-108, wherein the second antigen-binding domain is as depicted in Figure 20, Figure 17B, Figure 17C, Figure 17D, or Figure 17E.

109. The cell engager of claim 108, wherein said second antigen binding domain comprises SEQ ID NOs: 263 and 264.

110. The cell engager of claim 108, wherein said second antigen binding domain comprises SEQ ID NOs: 265 and 266.

111. The cell engager of any one of claims 100-110, wherein the cell engager comprises a third antigen-binding domain.

112. The cell engager of claim 111, wherein said third antigen-binding domain binds to a polypeptide expressed on the surface of a NK cell.

113. The cell engager of claim 112, wherein the polypeptide expressed on the surface of the NK cell is a CD16a, NKG2A, NKG2D, NKp30, NKp44, or NKp46 polypeptide.

114. The cell engager of claim 111, wherein the third antigen-binding domain is the antigen-binding domain depicted in Figure 19.

115. 15. A method for binding a molecule to a CD94 / NKG2A polypeptide on the surface of a NK cell or a T cell without inhibiting the cytotoxic activity of the NK cell or the T cell, comprising contacting the NK cell or the T cell with a composition of any one of claims 90-94 or a cell engager of any one of claims 100-114.

116. 116. The method of claim 115, wherein the method is in vitro.

117. 116. The method of claim 115, wherein the method is in vivo.

118. 118. The method of claim 117, wherein the method is in vivo in a mammal.

119. 119. The method of claim 118, wherein the mammal is a human.

120. 130. A method for reducing expression of a CD94 / NKG2A polypeptide on the surface of a NK cell or a T cell, comprising contacting said NK cell or said T cell with a composition of any one of claims 95-99 or a cell engager of any one of claims 100-114.

121. 121. The method of claim 120, wherein the method is in vitro.

122. 121. The method of claim 120, wherein the method is in vivo.

123. 123. The method of claim 122, wherein the method is in vivo in a mammal.

124. 124. The method of claim 123, wherein the mammal is a human.

125. HER2 + 130. A method of killing cancer cells, comprising contacting an immune cell positive for a CD94 / NKG2A polypeptide with (a) the composition of any one of claims 90-99 or the cell engager of any one of claims 100-114, and (b) an anti-HER2 antibody.

126. 126. The method of claim 125, wherein the method is in vitro.

127. 126. The method of claim 125, wherein the method is in vivo.

128. 128. The method of claim 127, wherein the method is in vivo in a mammal.

129. 129. The method of claim 128, wherein the mammal is a human.

130. The method of any one of claims 125 to 129, wherein the anti-HER2 antibody is pertuzumab.

131. 130. A method of treating a mammal having cancer comprising administering to said mammal a composition of any one of claims 90-99 or a cell engager of any one of claims 100-114.

132. 132. The method of claim 131, wherein the mammal is a human.

133. The cancer is HER2 + The method of any one of claims 131 to 132, wherein the cancer is cancer.

134. The method of claim 133, wherein the method further comprises administering an anti-HER2 antibody to the mammal.

135. The method of claim 134, wherein the anti-HER2 antibody is pertuzumab.

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