Anti-naprt antibodies and methods of use

Abstract

The present disclosure provides antibodies and polypeptides that specifically bind to nicotinate phosphoribosyltransferase (NAPRT). Also provided are nucleic acids encoding these antibodies, expression vectors and host cells for making these antibodies, methods of detecting NAPRT in a sample from a subject in need thereof using these antibodies, methods of diagnosing a subject with cancer or sepsis using these antibodies. Also provided are compositions comprising these antibodies. Also provided are compositions comprising a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi) and methods of treating cancer using an NAMPTi, after diagnosis using the NAPRT antibodies.

Description

ANTI-NAPRT ANTIBODIES AND METHODS OF USECROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to United States Provisional Patent Application serial number 63 / 598,869, filed November 14, 2023, the contents of which are hereby incorporated by reference in their entirety.FIELD

[0002] The present disclosure relates to antibodies that are specific for human nicotinate phosphoribosyltransferase (NAPRT), and methods of use thereof.BACKGROUND

[0003] The nicotinate phosphoribosyltransferase (NAPRT) gene has gained relevance in the research of cancer therapeutic strategies due to its main role as a nicotinamide adenine dinucleotide (NAD) biosynthetic enzyme. NAD metabolism is an attractive target for the development of anticancer therapies, given the high energy requirements of proliferating cancer cells and NAD- dependent signaling. Lack of NAPRT expression was observed in several cancer types and associated with NAPRT epigenetic silencing in some cases, such as gastric and lung cancer. On the other hand, NAPRT amplifications and overexpression were reported in ovarian, breast and pancreatic cancer. Differences in NAPRT expression between subtypes of cancer, namely in breast, pancreatic, lung and gastric carcinomas, suggest that individual variability should be considered in therapeutic approaches.

[0004] Thus, there is a need for antibodies specifically binding to NAPRT.SUMMARY

[0005] The present disclosure provides antibodies and polypeptides that specifically bind to NAPRT (e.g., human NAPRT). Also provided are pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for making these antibodies, and methods of detecting NAPRT in a sample from a subject using these antibodies. The anti-NAPRT antibodies provided herein are also useful in indicating the likelihood of a subject with cancer to be effectively treated with a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi). The anti-NAPRT antibodies provided herein are also useful in indicating the likelihood of a subject to have sepsis.

[0006] In one aspect, provided herein is an antibody that specifically binds human NAPRT, the antibody comprising: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NO: 1; and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NO: 2

[0007] In certain embodiments, the antibody comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13; 15, 16, and 5; or 17, 18, and 19, respectively.

[0008] In certain embodiments, the antibody comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21, and 8, respectively.

[0009] In certain embodiments, the antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 3, 4, 5, 6, 7, and 8; 9, 10, 5, 6, 7, and 8; 11, 12, 13, 14, LM, and 8; 11, 12, 13, 14, LMS, and 8; 15, 16, 5, 6, 7, and 8; 17, 18, 19, 20, 21, and 8, respectively.

[0010] In certain embodiments, the VH comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 1 and wherein the VL comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 2.

[0011] In certain embodiments, the antibody comprises the VH amino sequence and the VL amino acid sequence set forth in SEQ ID NOs: 1 and 2, respectively.

[0012] In certain embodiments, the antibody comprises a heavy chain constant region, optionally selected from the group consisting of human IgGi, IgG2, IgGs, IgG4, IgAi, and IgA2.

[0013] In certain embodiments, the antibody comprises a heavy chain constant region that is a variant of a wild-type heavy chain constant region, wherein the variant heavy chain constant region binds to an FcyR with lower affinity than the wild-type heavy chain constant region binds to the FcyR.

[0014] In another aspect, provided herein is an antibody that cross-competes for binding to NAPRT with any of the anti-NAPRT antibodies described herein.

[0015] In another aspect, provided herein is an antibody that specifically binds to the amino acid sequence of SEQ ID NO: 29 or 30.

[0016] In another aspect, provided herein is polypeptide comprising a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13; 15, 16, and 5; or 17, 18, and 19, respectively.

[0017] In another aspect is provided a polypeptide comprising a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NO: 2. In certain embodiments, the VL comprises the CDRL1, CDRL2, and CDRL3 set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21, and 8, respectively.

[0018] In another aspect, provided herein is a polypeptide comprising the amino acid sequence LM, LMS, or the amino acid sequences set forth in any one of SEQ ID NOs: 1-21.

[0019] In another aspect, provided herein is a polynucleotide encoding: a VH, a VL, a heavy chain, and / or a light chain of any of the antibodies described herein; or any of the polypeptides described herein.

[0020] In another aspect, provided herein is a vector comprising any of the polynucleotides described herein.

[0021] In another aspect, provided herein is a recombinant host cell comprising: a polynucleotide described herein, a vector described herein, a first polynucleotide encoding a heavy chain variable region or a heavy chain of an antibody described herein and a second polynucleotide encoding a light chain variable region or a light chain of an antibody described herein; and / or a first vector comprising a first polynucleotide encoding a heavy chain variable region or a heavy chain of an antibody described herein and a second vector comprising a second polynucleotide encoding a light chain variable region or a light chain of an antibody described herein.

[0022] In another aspect, provided herein is a composition comprising an antibody described herein, a polypeptide described herein, a polynucleotide described herein, a vector described herein, or a host cell described herein, and a pharmaceutically acceptable carrier or excipient.

[0023] In another aspect, provided herein is a method of producing an antibody, the method comprising culturing a host cell described herein under suitable conditions such that the polynucleotide is expressed, and the antibody is produced.

[0024] In another aspect, provided herein is a method of determining whether a sample comprising cells from a subject is nicotinic acid phosphoribosyltransferase (NAPRT) negativecomprising administering an antibody that specifically binds to NAPRT in the sample and detecting specific binding of the antibody to NAPRT in less than 1% of the cells in the sample, thereby determining that the sample is NAPRT negative.

[0025] In certain embodiments, the sample comprises cancer cells. In certain embodiments, the cancer cells are solid tumor cells. In certain embodiments, the cancer cells are thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer cells.

[0026] In another aspect, provided herein is a method of treating cancer comprising administering a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi) to a subject in need thereof, wherein a sample from the cancer has lower NAPRT levels than a control sample.

[0027] In certain embodiments, the method further comprises administering niacin to the subject.

[0028] In certain embodiments, the cancer is NAPRT negative.

[0029] In certain embodiments, the cancer is thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer.

[0030] In another aspect is provided a method of diagnosing a subject with a cancer with increased likelihood to be effectively treated with a NAMPTi comprising administering an antibody that specifically binds to NAPRT to a sample from the subject and detecting specific binding of the antibody to NAPRT, wherein if there is less binding to NAPRT in the sample from the subject than in a control sample, the cancer in the subject has an increased likelihood to be effectively treated with the NAMPTi.

[0031] In certain embodiments, the cancer in the subject has an increased likelihood to be effectively treated with niacin and the NAMPTi.

[0032] In certain embodiments, the cancer is thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma,mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer cells.

[0033] In another aspect, provided herein is a method of diagnosing a subject with sepsis comprising administering an antibody that specifically binds to NAPRT to a sample from the subject and detecting specific binding of the antibody to NAPRT, wherein if there is more binding to NAPRT in the sample from the subject than in a control sample, the subject has an increased likelihood to have sepsis.

[0034] In certain embodiments, the sample from the subject is NAPRT negative.

[0035] In certain embodiments, the NAPRT comprises the amino acid sequence of SEQ IDNO: 29 or 30.

[0036] In certain embodiments, the antibody comprises: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NO: 1; and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NO: 2.

[0037] In certain embodiments, the antibody comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13; 15, 16, and 5; or 17, 18, and 19, respectively.

[0038] In certain embodiments, the antibody comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21, and 8, respectively.

[0039] In certain embodiments, the antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 3, 4, 5, 6, 7, and 8; 9, 10, 5, 6, 7, and 8; 11, 12, 13, 14, LM, and 8; 11, 12, 13, 14, LMS, and 8; 15, 16, 5, 6, 7, and 8; 17, 18, 19, 20, 21, and 8, respectively.

[0040] In certain embodiments, the VH comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 1 and wherein the VL rises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NOs: 2.

[0041] In certain embodiments, the antibody comprises a heavy chain constant region, selected from the group consisting of human IgGi, IgG?, IgGs, IgG4, IgAi, and IgA2.

[0042] In certain embodiments, the antibody comprises a heavy chain constant region that is a variant of a wild-type heavy chain constant region, wherein the variant heavy chain constant region binds to an FcyR with lower affinity than the wild-type heavy chain constant region binds to the FcyR.BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG. 1 presents Western blot experiments showing the binding of two hybridoma clones to human NAPRT.

[0044] FIGS. 2A-2D present exemplary data showing the effectiveness of different NAMPTi of the present application in two patient-derived colorectal adenocarcinoma ex vivo models, one of which is NAPRT -positive and the other of which is NAPRT -negative. Following the excision of implanted tumors, tissue was either cryopreserved for later use or used immediately. Tumor samples were dissociated into single cells and smaller fragments by enzymatic digestion, filtered to remove large fragments, and then treated with increasing concentrations of NAMPTi in the presence of nicotinic acid. FIGS. 2A-2C are graphs showing the in vitro effects of three different NAMPTi, ATN-249 (FIG. 2A), ATN-940 (FIG. 2B) and ATN-1083 (FIG. 2C) in the NAPRT- Negative and NAPRT-positive patient-derived colorectal cancer ex vivo models, and their respective EC50 values (96h exposure, 0.1% final DMSO v / w; endpoint read using Cell Titer Glow 2.0 for viability via ATP detection). FIG. 2D shows NAPRT immunohistochemistry staining and negative background (Ms mAb) staining of two patient-derived ex vivo colorectal cancer models, (NAPRT -negative) and (NAPRT-positive).

[0045] FIGS. 3A-3D present exemplary data showing the in vitro effectiveness of the NAMPTi ATN-249 across various patient-derived tumor models (NAPRT-positive in FIGS. 3A and 3B, and NAPRT negative in FIGS. 3C and 3D). The NAPRT status was identified by Immunohistochemistry (IHC) and western blot (WB) using anti-NAPRT clone 4A5D7. Each graph includes a dotted line indicating 55% cytotoxicity, which was used to define whether a model responded to ATN-249.

[0046] FIG. 4 is a graph presenting exemplary in vivo data showing that NAMPTi ATN-940 led to strong inhibition of a patient-derived NAPRT-negative tumor, while having minimal effect on tumor growth of a patient-derived NAPRT-positive tumor.

[0047] FIG. 5 is a table providing details on the different ex vivo models used in the experiments shown in FIGS. 2A-2D.DETAILED DESCRIPTION

[0048] The instant disclosure provides anti-NAPRT antibodies and polypeptides that specifically binds NAPRT. Also provided are pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for making these antibodies, and methods of detecting NAPRT in sample from a subject using these antibodies. In some embodiments, the anti-NAPRT antibodies provided herein indicate the likelihood of a subject with cancer to be effectively treated with a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi). In some embodiments, when the cells in a sample are NAPRT negative this indicates an increased likelihood of a subject with cancer to be effectively treated with a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi). In some embodiments, the anti-NAPRT antibodies indicate the likelihood of a subject to have sepsis.Definitions

[0049] As used herein, the term “NAPRT,” refers to nicotinic acid phosphoribosyltransferase, an enzyme that catalyzes the first step in the biosynthesis of NAD from nicotinic acid, the ATP- dependent synthesis of beta-nicotinate D -ribonucleotide from nicotinate and 5-phospho-D-ribose 1 -pyrophosphate. As used herein, the term "human NAPRT" refers to a protein encoded by a wildtype human NAPRT gene (e.g., the gene set forth in Genbank accession number NM_145201 .6, or NCBI Gene ID: 93100). The amino acid sequence of an exemplary human NAPRT protein is set forth in NCBI Reference Sequence: NP 660202.3. In some embodiments, human NAPRT protein comprises the amino acid sequence of SEQ ID NO: 31. In some embodiments, the human NAPRT comprises the sequence set forth in UniProtKB accession number Q6XQN6-1. In some embodiments, the human NAPRT comprises the sequence set forth in NCBI Reference Sequence: NP 660202.3. In some embodiments, the canonical amino acid sequence human NAPRT comprises the amino acid sequence of SEQ ID NO: 36. In some embodiments, the human NAPRT comprises the sequence set forth in UniProtKB accession number Q6XQN6-2. In some embodiments, the functional human NAPRT comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the human NAPRT comprises the sequence set forth in UniProtKB accession number Q6XQN6-3. In some embodiments, the human NAPRT comprises the sequence set forth in NCBI Reference Sequence: NP_001273758.1. In some embodiments, the functionalhuman NAPRT comprises the amino acid sequence of SEQ ID NO: 38. In some embodiments, the functional human NAPRT comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, NAPRT, comprises the amino acid sequence of any one of SEQ ID NOs: 29-40.

[0050] In some embodiments, the NAPRT is selected from Table 1 below.Table 1: Exemplary NAPRT variants

[0051] As used herein, the term “detectable NAPRT” refers to NAPRT that can be detected in a sample using one of the anti-NAPRT antibodies or polypeptides disclosed herein.

[0052] As used herein, the term “NAPRT negative” regarding a sample comprising cells means that less than 1% of the cells in the sample have detectable NAPRT. As used herein, the term “NAPRT negative” regarding a cancer means that less than 1% of the cells in a sample from the cancer have detectable NAPRT. In some embodiments, NAPRT that can be detected in a sample is by using one of the anti-NAPRT antibodies or polypeptides disclosed herein.

[0053] As used herein, the terms “antibody” and “antibodies” include full-length antibodies, antigen-binding fragments of full-length antibodies, and molecules comprising antibody CDRs, VH regions, and / or VL regions. Examples of antibodies include, without limitation, monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, an antibody light chain monomer, an antibody heavy chain monomer, an antibody light chain dimer, an antibody heavy chain dimer, an antibody light chain-antibody heavy chain pair, intrabodies, heteroconjugate antibodies, antibody-drug conjugates, single domainantibodies, monovalent antibodies, single-chain antibodies or single-chain Fvs (scFv), camelized antibodies, affibodies, Fab fragments, F(ab’)2 fragments, disulfide-linked Fvs (sdFv), anti- idiotypic (anti-Id) antibodies (including, e.g., anti-anti-Id antibodies), and antigen -binding fragments of any of the above. In certain embodiments, antibodies described herein refer to polyclonal antibody populations. Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgGi, IgG2, IgG3, I G4, IgAi. or IgA2), or any subclass (e.g., IgG2a or IgG2b) of immunoglobulin molecule. In certain embodiments, antibodies described herein are IgG antibodies, or a class (e.g., human IgGi or IgG4) or subclass thereof. In a specific embodiment, the antibody is a humanized monoclonal antibody. In another specific embodiment, the antibody is a human monoclonal antibody.

[0054] “Multispecific antibodies” are antibodies (e.g., bispecific antibodies) that specifically bind to two or more different antigens or two or more different regions of the same antigen. Multispecific antibodies include bispecific antibodies that contain two different antigen-binding sites (exclusive of the Fc region). Multispecific antibodies can include, for example, recombinantly produced antibodies, human antibodies, humanized antibodies, resurfaced antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, an antibody light chain monomer, heteroconjugate antibodies, linked single-chain antibodies or linked-single-chain Fvs (scFv), camelized antibodies, affybodies, linked Fab fragments, F(ab’)2 fragments, chemically-linked Fvs, and disulfide-linked Fvs (sdFv). Multi specific antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgGi, IgG2, IgG3, IgG4, IgAi, or IgA2), or any subclass (e.g., IgG2a or IgG2b) of immunoglobulin molecule. In certain embodiments, multispecific antibodies described herein are IgG antibodies, or a class (e.g., human IgGi, IgG2, or IgG4) or subclass thereof.

[0055] As used herein, the term “CDR” or “complementarity determining region” means the noncontiguous antigen combining sites found within the variable regions of heavy and light chain polypeptides. These particular regions have been described by, for example, Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991), by Chothia et al., J. Mol. Biol. 196: 901-917 (1987), and by MacCallum et al., J. Mol. Biol. 262: 732-745 (1996), all of which are herein incorporated by reference in their entireties, where the definitions include overlapping or subsets of amino acid residues when compared against each other. In certain embodiments, the term “CDR” is a CDR as defined by MacCallumet al., J. Mol. Biol. 262:732-745 (1996) and Martin A. “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Diibel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001). In certain embodiments, the term “CDR” is a CDR as defined by Kabat etal., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat etal., Sequences of protein of immunological interest. (1991). In certain embodiments, heavy chain CDRs and light chain CDRs of an antibody are defined using different conventions. In certain embodiments, heavy chain CDRs and / or light chain CDRs are defined by performing structural analysis of an antibody and identifying residues in the variable region(s) predicted to make contact with an epitope region of a target molecule (e.g. , human NAPRT). CDRH1 , CDRH2, and CDRH3 denote the heavy chain CDRs, and CDRL1, CDRL2, and CDRL3 denote the light chain CDRs.

[0056] As used herein, the terms “variable region” and “variable domain” are used interchangeably and are common in the art. The variable region typically refers to a portion of an antibody, generally, a portion of a light or heavy chain, typically about the amino-terminal 110 to 120 amino acids or 110 to 125 amino acids in the mature heavy chain and about 90 to 115 amino acids in the mature light chain, which differ extensively in sequence among antibodies and are used in the binding and specificity of a particular antibody for its particular antigen. The variability in sequence is concentrated in those regions called complementarity determining regions (CDRs) while the more highly conserved regions in the variable region are called framework regions (FRs). Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of the antibody with antigen. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and human framework regions (FRs). In certain embodiments, the variable region is a primate (e.g., non-human primate) variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0057] As used herein, the terms “VH” and “VL” refer to antibody heavy and light chain variable regions, respectively, as described in Kabat et al., (1991) Sequences of Proteins of Immunological Interest (NTH Publication No. 91-3242, Bethesda), which is herein incorporated by reference in its entirety.

[0058] As used herein, the term “constant region” is common in the art. The constant region is an antibody portion, e.g., a carboxyl terminal portion of a light and / or heavy chain, which is notdirectly involved in binding of an antibody to antigen but which can exhibit various effector functions, such as interaction with an Fc receptor (e.g., Fc gamma receptor).

[0059] As used herein, the term “heavy chain” when used in reference to an antibody can refer to any distinct type, e.g., alpha (a), delta (6), epsilon (s), gamma (y), and mu (p), based on the amino acid sequence of the constant region, which give rise to IgA, IgD, IgE, IgG, and IgM classes of antibodies, respectively, including subclasses of IgG, e.g., IgGi, IgG2, IgGs, and IgG- .

[0060] As used herein, the term “light chain” when used in reference to an antibody can refer to any distinct type, e.g. , kappa (K) or lambda (A), based on the amino acid sequence of the constant region. Light chain amino acid sequences are well known in the art. In specific embodiments, the light chain is a human light chain.

[0061] As used herein, the term “specifically binds” refers to the specificity of a binding molecule (e.g., an antibody) for an antigen, as is understood by one skilled in the art. Binding molecules that specifically bind to an antigen typically bind to the antigen with an equilibrium dissociation constant (KD) of less than 1 x 106M, as measured by, e.g., ELISA assay, surface plasmon resonance, or other suitable assays known in the art. The skilled worker will appreciate that, in certain embodiments, a binding molecule can specifically bind to different antigens, e.g., different antigens that share a common epitope that is recognized by the binding molecule.

[0062] As used herein, the term “affinity” refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity which reflects a 1 : 1 interaction between members of a binding pair (e.g. , antibody and antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (KD). Affinity can be measured by common methods known in the art, including those described herein.

[0063] As used herein, the term “EU numbering system” refers to the EU numbering convention for the constant regions of an antibody, as described in Edelman, G.M. et al., Proc. Natl. Acad. USA, 63, 78-85 (1969) and Kabat et al, Sequences of Proteins of Immunological Interest, U.S. Dept. Health and Human Services, 5th edition, 1991, each of which is herein incorporated by reference in its entirety.

[0064] As used herein, the term “treat,” “treating,” and “treatment” refer to therapeutic or preventative measures described herein. The methods of “treatment” employ administration of anantibody to a subject having a disease or disorder, or predisposed to having such a disease or disorder, in order to prevent, cure, delay, reduce the severity of, or ameliorate one or more symptoms of the disease or disorder or recurring disease or disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.

[0065] As used herein, an “inhibitor” refers to a compound (e.g., compounds described herein) that reduces activity when compared to a control, such as absence of the compound or a compound with known inactivity.

[0066] As defined herein, the term “inhibition”, “inhibit”, “inhibiting” and the like in reference to a protein- inhibitor interaction means negatively affecting (e.g., decreasing) the activity or function of the protein relative to the activity or function of the protein in the absence of the inhibitor. In embodiments inhibition means negatively affecting (e.g., decreasing) the concentration or levels of the protein relative to the concentration or level of the protein in the absence of the inhibitor. In embodiments inhibition refers to reduction of a disease or symptoms of disease. In embodiments, inhibition refers to a reduction in the activity of a particular protein target. Thus, inhibition includes, at least in part, partially or totally blocking stimulation, decreasing, preventing, or delaying activation, or inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity or the amount of a protein. In embodiments, inhibition refers to a reduction of activity of a target protein resulting from a direct interaction (e.g., an inhibitor binds to the target protein). In embodiments, inhibition refers to a reduction of activity of a target protein from an indirect interaction (e.g., an inhibitor binds to a protein that activates the target protein, thereby preventing target protein activation).

[0067] The terms “inhibitor,” “repressor” or “antagonist” or “downregulator” interchangeably refer to a substance capable of detectably decreasing the expression or activity of a given gene or protein. The antagonist can decrease expression or activity 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in comparison to a control in the absence of the antagonist. In certain instances, expression or activity is 1.5 -fold, 2-fold, 3 -fold, 4-fold, 5 -fold, 10-fold or lower than the expression or activity in the absence of the antagonist.

[0068] The terms “niacin” or “vitamin B3” refer to one of the water-soluble B vitamins. Niacin is the generic name for nicotinic acid (pyridine-3 -carboxylic acid), nicotinamide (niacinamide or pyridine-3-carboxamide), and related derivatives, such as nicotinamide riboside. Niacin isnaturally present in many foods, added to some food products, and available as a dietary supplement.

[0069] The terms “NAMPT” or “nicotinamide phosphoribosyltransferase” refer to an enzyme encoded by a wild-type human NAMPT gene (e.g., the gene set forth in Genbank accession number NM_005746, or NCBI Gene ID; 10135). This gene encodes a protein that catalyzes the condensation of nicotinamide with 5 -phosphoribosyl- 1 -pyrophosphate to yield nicotinamide mononucleotide, one step in the biosynthesis of nicotinamide adenine dinucleotide. The protein belongs to the nicotinic acid phosphoribosyltransferase (NAPRTase) family and is thought to be involved in many important biological processes, including metabolism, stress response and aging. The amino acid sequence of an exemplary human NAPRT protein is set forth in NCBI Reference Sequence: NP_660202.3.

[0070] The term “NAMPTi” or “nicotinamide phosphoribosyltransferase inhibitor” refers to a compound capable of inhibiting or decreasing the activity of NAMPT. In some embodiments, the NAMPTi is FK866 (PMID; 14612543). In some embodiments, the NAMPTi is GNE617 (PMID: 24403854). In some embodiments, the NAMPTi is OT82 (PMID; 31896781). In some embodiments, the NAMPTi is LSN3154567 (PMID; 29054982).

[0071] The terms “disease” or “condition” refer to a state of being or health status of a patient or subject capable of being treated with the compounds or methods provided herein. The disease may be a cancer. The disease may be an autoimmune disease. The disease may be an inflammatory disease. The disease may be an infectious disease.

[0072] As used herein, the term "cancer" refers to all types of cancer, neoplasm or malignant tumors found in mammals (e.g., humans), including thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer.

[0073] In some embodiments, NAPRT is not detected in samples obtained from certain cancers or tumors, using one of the antibodies or polypeptides disclosed herein. In some embodiments, no NAPRT is detected in samples from thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer. In some embodiments, no NAPRT is detect in 50% to 100% of the samples obtained from thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lungcancer (SCLC), hepatocellular carcinoma, mesothelioma. In some embodiments, no NAPRT is detect in 13% to 100% of the samples obtained from thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer.

[0074] As used herein, the terms “sepsis”, “septicemia”, “septicaemia”, or “blood poisoning”, refers to a potentially life-threatening condition resulting from the presence of harmful microorganisms in the blood or other tissues and the body’s response to their presence, potentially leading to the malfunctioning of various organs, shock, and death. Sepsis occurs when chemicals released in the bloodstream to fight an infection trigger inflammation throughout the body. This can cause a cascade of changes that damage multiple organ systems, leading them to fail, sometimes even resulting in death.

[0075] As used herein, the term “effective amount” in the context of the administration of a therapy to a subject refers to the amount of a therapy that achieves a desired prophylactic or therapeutic effect.

[0076] As used herein, the term “subject” includes any human or non-human animal. In certain embodiments, the subject is a human or non-human mammal. In certain embodiments, the subject is a human.

[0077] As used herein, the term “control sample” is used in accordance with its plain ordinary meaning and refers to a sample obtained from a healthy subject. In some embodiments, the control sample is obtained from a subject that does not have cancer. In some embodiments, the control sample is obtained from a subject that does not have sepsis. In some embodiments, NAPRT is detected in a control sample obtained from a healthy subject. In some embodiments, NAPRT is detected in a control sample obtained from a subject that does not have cancer. In some embodiments, NAPRT is detected in a control sample obtained from a subject that does not have sepsis.

[0078] As used herein with respect to an antibody or polynucleotide, the term “isolated” refers to an antibody or polynucleotide that is separated from one or more contaminants (e.g., polypeptides, polynucleotides, lipids, or carbohydrates, etc.) which are present in a natural source of the antibody or polynucleotide. All instances of “isolated antibodies” described herein areadditionally contemplated as antibodies that may be, but need not be, isolated. All instances of “isolated polynucleotides” described herein are additionally contemplated as polynucleotides that may be, but need not be, isolated. All instances of “antibodies” described herein are additionally contemplated as antibodies that may be, but need not be, isolated. All instances of “polynucleotides” described herein are additionally contemplated as polynucleotides that may be, but need not be, isolated.

[0079] The determination of “percent identity” between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be accomplished using a mathematical algorithm. A specific, non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87: 2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90: 5873-5877, each of which is herein incorporated by reference in its entirety. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul SF et al., (1990) J Mol Biol 215: 403, which is herein incorporated by reference in its entirety. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecule described herein. BLAST protein searches can be performed with the XBLAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, which is herein incorporated by reference in its entirety. Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id ). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4: 11-17, which is herein incorporated by reference in its entirety. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM 120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0080] The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.Anti-NAPRT Antibodies

[0081] In one aspect, the instant disclosure provides antibodies that specifically bind to NAPRT (e.g., human NAPRT). The VH, VL, CDRH and CDRL amino acid sequences of exemplary antibodies are set forth in Tables 2, 3, and 4 below.Table 2. VH and VL amino acid sequences of an exemplary anti-NAPRT antibody.Table 3. CDRH amino acid sequences of exemplary anti-NAPRT antibody of Table 2, according to different numbering schemes.Table 4. CDRL amino acid sequences of exemplary anti-NAPRT antibody of Table 2, according to different numbering schemes.

[0082] The individual CDRs of an antibody disclosed herein can be determined according to any CDR numbering scheme known in the art.

[0083] In certain embodiments, one or more of the CDRs of an antibody disclosed herein can be determined according to Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest (1991), each of which is herein incorporated by reference in its entirety.

[0084] In certain embodiments, an antibody provided herein comprises the CDRH1, CDRH2, and / or CDRH3 of a VH amino acid sequence set forth in SEQ ID NO: 1 as determined by the Kabat numbering scheme. In certain embodiments, an antibody provided herein comprises the CDRL1, CDRL2, and / or CDRL3 of a VL amino acid sequence set forth in SEQ ID NO: 2 as determined by the Kabat numbering scheme.

[0085] In certain embodiments, one or more of the CDRs of an antibody disclosed herein can be determined according to the Chothia numbering scheme, which refers to the location of immunoglobulin structural loops (see, e.g., Chothia C & Lesk AM, (1987), J Mol Biol 196: 901 - 917; Al-Lazikani B et al., (1997) J Mol Biol 273: 927-948; Chothia C et al., (1992) J Mol Biol 227: 799-817; Tramontane A et al., (1990) J Mol Biol 215(1): 175-82; and U.S. Patent No. 7,709,226, all of which are herein incorporated by reference in their entireties).

[0086] In certain embodiments, an antibody provided herein comprises the CDRH1, CDRH2, and / or CDRH3 of a VH amino acid sequence set forth in SEQ ID NO: 1 as determined by the Chothia numbering scheme. In certain embodiments, an antibody provided herein comprises the CDRL1, CDRL2, and / or CDRL3 of a VL amino acid sequence set forth in SEQ ID NO; 2 as determined by the Chothia numbering system.

[0087] In certain embodiments, one or more of the CDRs of an antibody disclosed herein can be determined according to MacCallum RM et al., (1996) J Mol Biol 262: 732-745, herein incorporated by reference in its entirety. See also, e.g., Martin A. “Protein Sequence and StructureAnalysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Diibel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001), herein incorporated by reference in its entirety.

[0088] In certain embodiments, an antibody provided herein comprises the CDRH1, CDRH2, and / or CDRH3 of a VH amino acid sequence set forth in SEQ ID NO; 1 as determined by the MacCallum numbering scheme. In certain embodiments, an antibody provided herein comprises the CDRL1, CDRL2, and / or CDRL3 of a VL amino acid sequence set forth in SEQ ID NO; 2 as determined by the MacCallum numbering system.

[0089] In certain embodiments, the CDRs of an antibody disclosed herein can be determined according to the IM GT numbering system as described in: Lefranc M-P, (1999) The Immunologist 7: 132-136; Lefranc M-P et al., (1999) Nucleic Acids Res 27: 209-212, each of which is herein incorporated by reference in its entirety; and Lefranc M-P et al., (2009) Nucleic Acids Res 37: D1006-D1012.

[0090] In certain embodiments, an antibody provided herein comprises the CDRH1, CDRH2, and / or CDRH3 of a VH amino acid sequence set forth in SEQ ID NO: 1 as determined by the IMGT numbering scheme. In certain embodiments, an antibody provided herein comprises the CDRL1, CDRL2, and / or CDRL3 of a VL amino acid sequence set forth in SEQ ID NO: 2 as determined by the IMGT numbering system.

[0091] In certain embodiments, the CDRs of an antibody disclosed herein can be determined according to the AbM numbering scheme, which refers to AbM hypervariable regions, which represent a compromise between the Kabat CDRs and Chothia structural loops and are used by Oxford Molecular’s AbM antibody modeling software (Oxford Molecular Group, Inc.), herein incorporated by reference in its entirety.

[0092] In certain embodiments, an antibody provided herein comprises the CDRH1, CDRH2, and / or CDRH3 of a VH amino acid sequence set forth in SEQ ID NO: 1 as determined by the AbM numbering scheme. In certain embodiments, an antibody provided herein comprises the CDRL1, CDRL2, and / or CDRL3 of a VL amino acid sequence set forth in SEQ ID NO; 2 as determined by the AbM numbering scheme.

[0093] In certain embodiments, the CDRs of an antibody disclosed herein can be determined according to the AHo numbering system, as described in Honegger and Pliickthun, A., J. Mol. Biol. 309:657-670 (2001), herein incorporated by reference in its entirety.

[0094] In certain embodiments, an antibody provided herein comprises the CDRH 1 , CDRH2, and / or CDRH3 of a VH amino acid sequence set forth in SEQ ID NO: 1 as detemiined by the AHo numbering scheme. In certain embodiments, an antibody provided herein comprises the CDRL1, CDRL2, and / or CDRL3 of a VL amino acid sequence set forth in SEQ ID NO: 2 as determined by the AHo numbering system.

[0095] In certain embodiments, the individual CDRs of an antibody disclosed herein are each independently determined according to one of the Kabat, Chothia, MacCallum, IMGT, AHo, or AbM numbering schemes, or by structural analysis of the multispecific molecule, wherein the structural analysis identifies residues in the variable region(s) predicted to make contact with an epitope region of NAPRT.

[0096] In certain embodiments, the instant disclosure provides an antibody that specifically binds NAPRT (e.g, human NAPRT) comprising a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of a VH amino acid sequence set forth in SEQ ID NO: 1, and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of a VL amino acid sequence set forth in SEQ ID NO: 2, wherein each CDR is independently determined according to one of the Kabat, Chothia, MacCallum, IMGT, AHo, or AbM numbering schemes, or by structural analysis of the multispecific molecule, wherein the structural analysis identifies residues in the variable region(s) predicted to make contact with an epitope region of NAPRT (e.g., human NAPRT).

[0097] In certain embodiments, the instant disclosure provides an antibody that specifically binds NAPRT (e.g., human NAPRT) comprising a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of a VH amino acid sequence set forth in SEQ ID NO: 1, and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of a VL amino acid sequence set forth in SEQ ID NOs: 2, wherein the VH amino acid sequence and the VL amino acid sequence are the same (i.e., both VH’s comprise the CDRH1, CDRH2, and CDRH3 amino acid sequences of a VH amino acid sequence set forth in SEQ ID NO: 1 , and both VH’s comprise a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of a VL amino acid sequence set forth in SEQ ID NOs: 2), wherein each CDR is independently determined according to one of the Kabat, Chothia, MacCallum, IMGT, AHo, or AbM numbering schemes, or by structural analysis of the multispecific molecule, wherein the structural analysis identifies residues in the variable region(s) predicted to make contact with an epitope region of NAPRT (e.g., human NAPRT).

[0098] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), wherein the antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences of the VH and VL amino acid sequences set forth in SEQ ID NOs: 1 and 2, respectively.

[0099] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), wherein the antibody comprises a VH comprising a CDRH1, CDRH2, and / or CDRH3 amino acid sequence set forth in Table 3. In certain embodiments, the antibody comprises a VH comprising a CDRH1 amino acid sequence set forth in any one of SEQ ID NOs: 3, 9, 11, 15, or 17. In certain embodiments, the antibody comprises a VH comprising a CDRH2 amino acid sequence set forth in any one of SEQ ID NOs: 4, 10, 12, 16, or 18. In certain embodiments, the antibody comprises a VH comprising a CDRH3 amino acid sequence set forth in any one of SEQ ID NOs: 5, 13, or 19.

[0100] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g. , human NAPRT), wherein the antibody comprises a VH comprising the CDRH1 , CDRH2, and CDRH3 amino acid sequences of any of the antibodies in Table 3. In certain embodiments, the antibody comprises a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13; 15, 16, and 5; or 17, 18, and 19, respectively.

[0101] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), wherein the antibody comprises a VL comprising a CDRL1, CDRL2, and / or CDRL3 amino acid sequence set forth in Table 4. In certain embodiments, the antibody comprises a VL comprising a CDRL1 amino acid sequence set forth in any one of SEQ ID NOs: 6, 14, or 20. In certain embodiments, the antibody comprises a VL comprising a CDRL2 amino acid sequence set forth in any one of SEQ ID NOs: 7, 21, LM, or LMS. In certain embodiments, the antibody comprises a VL comprising a CDRL3 amino acid sequence set forth in SEQ ID NO: 8.

[0102] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), wherein the antibody comprises a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of any of the antibodies in Table 4. In certain embodiments, the antibody comprises a VL comprising the CDRL1, CDRL2, and CDRL3 aminoacid sequences set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21 , and 8, respectively.

[0103] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), wherein the antibody comprises a VH comprising a CDRH1, CDRH2, and / or CDRH3 amino acid sequence set forth in Table 3 and a VL comprising a CDRL1, CDRL2, and / or CDRL3 amino acid sequence set forth in Table 4, wherein 2, 3, 4, 5, or 6 of the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences are from the same antibody (i.e., as shown in Tables 3 and 4). In certain embodiments, the antibody comprises a VH comprising CDRH1, CDRH2, and CDRH3 regions, and a VL comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4, 5, 6, 7, and 8; 9, 10, 5, 6, 7, and 8; 11, 12, 1314, LM, and 8; 11, 12, 13, 14, LMS, and 8; 15, 16, 5, 6, 7, and 8; or 17, 18, 19, 20, 21, and 8, respectively.

[0104] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT) comprising a VH comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), comprising a VH comprising the amino acid sequence set forth in SEQ ID NO:1. In certain embodiments, the amino acid sequence of the VH consists of the amino acid sequence set forth in SEQ ID NOs: 1.

[0105] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), comprising a VL comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g, at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to the amino acid sequence set forth in SEQ ID NO: 2. In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:2. In certain embodiments, the amino acid sequence of the VL consists of the amino acid sequence set forth in SEQ ID NO: 2.

[0106] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g. , human NAPRT), comprising a VH comprising an amino acid sequence thatis at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98 or 99%) identical to the amino acid sequence set forth in SEQ ID NO: 1, and a VL comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to the amino acid sequence set forth in SEQ ID NO: 2. In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), comprising a VH comprising the amino acid sequence of SEQ ID NO: 1, and a VL comprising the amino acid sequence of SEQ ID NO: 2. In certain embodiments, the amino acid sequence of the VH consists of the amino acid sequence set forth in SEQ ID NO: 1, and the amino acid sequence of the VL consists of the amino acid sequence set forth in SEQ ID NO: 2.

[0107] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g. , human NAPRT), comprising a VH comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%) identical to the amino acid sequence set forth in SEQ ID NO: 1, and a VL comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to the amino acid sequence set forth in SEQ ID NO: 2, wherein the amino acid sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2 are from the same antibody (i.e., as shown in Table 2). In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), comprising a VH comprising the amino acid sequence of SEQ ID NO: 1, and a VL comprising the amino acid sequence of SEQ ID NO: 2, wherein the amino acid sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2 are from the same antibody (i.e., as shown in Table 2). In certain embodiments, the amino acid sequence of the VH consists of the amino acid sequence set forth in SEQ ID NO: 1, and the amino acid sequence of the VL consists of the amino acid sequence set forth in SEQ ID NO: 2, wherein the amino acid sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2 are from the same antibody (i.e., as shown in Table 2).

[0108] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), wherein the antibody comprises a VH and a VL, wherein the VH comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 1 and wherein the VL rises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NOs: 2.

[0109] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e. ., human NAPRT), wherein the antibody comprises a VH and a VL comprising the amino acid sequences set forth in SEQ ID NOs: 1 and 2, respectively.

[0110] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g. , human NAPRT), wherein the antibody comprises a VH and a VL consisting of the amino acid sequences set forth in SEQ ID NOs: 1 and 2, respectively.

[0111] In certain embodiments, the instant disclosure provides an antibody that crosscompetes for binding to NAPRT (e.g., human NAPRT) with any of the antibodies described above. In certain embodiments, the instant disclosure provides an antibody that binds to the same or an overlapping epitope of NAPRT (e.g., an epitope of human NAPRT) as an antibody described above.

[0112] In certain embodiments, the instant disclosure provides an antibody that specifically binds to the amino acid sequences of SEQ ID NO: 29 or SEQ ID NO: 30. In certain embodiments, the instant disclosure provides an antibody that specifically binds to the amino acid sequences of SEQ ID NO: 29 and SEQ ID NO: 30. In certain embodiments, the instant disclosure provides an antibody that does not specifically bind to the amino acid sequence of SEQ ID NO: 28.

[0113] In certain embodiments, the epitope of an antibody can be determined by, e.g., NMR spectroscopy, surface plasmon resonance (BIAcore®), X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange coupled with mass spectrometry (e.g., liquid chromatography electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). For X-ray crystallography, crystallization may be accomplished using any of the known methods in the art (e.g., Giege R et al., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350; McPherson A (1990) Eur J Biochem 189: 1-23; ChayenNE (1997) Structure 5: 1269-1274; McPherson A (1976) J Biol Chem 251 : 6300-6303, all of which are herein incorporated by reference in their entireties). Antibody:antigen crystals may be studied using well known X-ray diffraction techniques and may be refined using computer software such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc.; see, e.g., Meth Enzymol (1985) volumes 114 & 115, eds. Wyckoff HW et al , U.S. Patent Application No. 2004 / 0014194), and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, ed Carter CW; Roversi P etal., (2000) Acta Crystallogr D Biol Crystallogr 56(Pt 10): 1316-1323,all of which are herein incorporated by reference in their entireties). Mutagenesis mapping studies may be accomplished using any method known to one of skill in the art. See, e.g., Champe M et al., (1995) supra and Cunningham BC & Wells JA (1989) supra for a description of mutagenesis techniques, including alanine scanning mutagenesis techniques. In a specific embodiment, the epitope of an antibody is determined using alanine scanning mutagenesis studies. In addition, or antibodies that recognize and bind to the same or overlapping epitopes of NAPRT (e.g., human NAPRT) can be identified using routine techniques such as an immunoassay, for example, by showing the ability of one antibody to block the binding of another antibody to a target antigen, i.e., a competitive binding assay. Competition binding assays also can be used to determine whether two antibodies have similar binding specificity for an epitope. Competitive binding can be determined in an assay in which the immunoglobulin under test inhibits specific binding of a reference antibody to a common antigen, such as NAPRT (e.g., human NAPRT). Numerous types of competitive binding assays are known, for example: solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see Stahli C et al., (1983) Methods Enzymol 9: 242-253); solid phase direct biotin-avidin EIA (see Kirkland TN et al., (1986) J Immunol 137: 3614-9); solid phase direct labeled assay, solid phase direct labeled sandwich assay (see Harlow E & Lane D, (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Press); solid phase direct label RIA using 1-125 label (see Morel GA et al., (1988) Mol Immunol 25(1): 7-15); solid phase direct biotinavidin EIA (see Cheung RC et al., (1990) Virology 176: 546-52); and direct labeled RIA (see Moldenhauer G et al., (1990) Scand J Immunol 32: 77-82), all of which are herein incorporated by reference in their entireties. Typically, such an assay involves the use of purified antigen (e.g., NAPRT, such as human NAPRT) bound to a solid surface or cells bearing either of these, an unlabeled test immunoglobulin and a labeled reference immunoglobulin. Competitive inhibition can be measured by determining the amount of label bound to the solid surface or cells in the presence of the test immunoglobulin. Usually, the test immunoglobulin is present in excess. Usually, when a competing antibody is present in excess, it will inhibit specific binding of a reference or antibody to a common antigen by at least 50-55%, 55-60%, 60-65%, 65-70%, 70- 75%, or more. A competition binding assay can be configured in a large number of different formats using either labeled antigen or labeled antibody. In a common version of this assay, the antigen is immobilized on a 96-well plate. The ability of unlabeled antibodies to block the bindingof labeled antibodies to the antigen is then measured using radioactive or enzyme labels. For further details see, e.g., Wagener C et al., (1983) J Immunol 130: 2308-2315; Wagener C et al., (1984) J Immunol Methods 68: 269-274; Kuroki M et al., (1990) Cancer Res 50: 4872-4879; Kuroki M et al., (1992) Immunol Invest 21 : 523-538; Kuroki M etal., (1992) Hybridoma 11 : 391- 407 and Antibodies: A Laboratory Manual, Ed Harlow E & Lane D editors supra, pp. 386-389, all of which are herein incorporated by reference in their entireties.

[0114] The anti-NAPRT antigen-binding molecules of the present disclosure can be linked to or co-expressed with another functional molecule, e.g., another peptide or protein. For example, an antibody or fragment thereof can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association, or otherwise) to one or more other molecular entities, such as another antibody or antibody fragment to produce a bispecific or a multispecific antibody with a second or additional binding specificity.

[0115] In certain embodiments, the antibody disclosed herein is conjugated to a radionuclide, or detectable label. In certain embodiments, the radionuclide is selected from the group consisting of the isotopes3H,14C,32P,35S,36C1,51Cr,57Co,58Co,59Fe,67Cu,90Y, "Tc,i nIn,117Lu,121I,124I,125I,131I,198Au,211At,213Bi,225AC, and186Re. In certain embodiments, the detectable label comprises a fluorescent moiety or a click chemistry handle.

[0116] Any immunoglobulin (Ig) constant region can be used in the antibodies disclosed herein. In certain embodiments, the Ig region is a human IgG, IgE, IgM, IgD, IgA, or IgY immunoglobulin molecule, any class (e.g., IgGi, IgG2, IgGs, IgG4, IgAi, and IgA2), or any subclass (e.g., IgG2a and IgG2b) of immunoglobulin molecule.

[0117] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), the antibody comprising a heavy chain constant region, optionally selected from the group consisting of human IgGi, IgG2, IgGs, IgG4, IgAi, and IgA2.

[0118] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), the antibody comprising a heavy chain constant region that is a variant of a wild-type heavy chain constant region, wherein the variant heavy chain constant region binds to an FcyR with lower affinity than the wild-type heavy chain constant region binds to the FcyR.

[0119] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), the antibody comprising a heavy chain constant regioncomprising an amino acid sequence shown in Table 5. In certain embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 24 or 25. In certain embodiments, the heavy chain constant region consists of the amino acid sequence of SEQ ID NO: 24 or 25.Table 5. Heavy chain constant region amino acid sequences of exemplary anti-NAPRT antibodies.

[0120] In certain embodiments, the instant disclosure provides an antibody that specifically binds to NAPRT (e.g., human NAPRT), the antibody comprising a light chain constant region comprising an amino acid sequence shown in Table 6. In certain embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 26 or 27. In certain embodiments, the heavy chain constant region consists of the amino acid sequence of SEQ ID NO: 26 or 27.Table 6. Light chain constant region amino acid sequences of exemplary anti-NAPRT antibodies.

[0121] In certain embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into an Fc region (e.g., a CH2 domain (residues 231-340 of human IgGi)) and / or a CH3 domain (residues 341-447 of human IgGi, numbered according to the EU numbering system) and / or a hinge region (residues 216-230, numbered according to the EU numbering system) of an antibody described herein, to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cellular cytotoxicity.

[0122] In certain embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of an antibody described herein, such that the number of cysteine residues in the hinge region is altered (e.g., increased or decreased) as described in, e.g., U.S. Patent No. 5,677,425, herein incorporated by reference in its entirety. The number of cysteine residues in the hinge region may be altered to, e.g., facilitate assembly of the light and heavy chains, or to alter (e.g., increase or decrease) the stability of the antibody.

[0123] In a specific embodiment, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into an IgG constant region, or FcRn-binding fragment thereof (preferably an Fc or hinge-Fc fragment) to alter (e.g., decrease or increase) half-life of the antibody in vivo. See, e.g., International Publication Nos. WO 02 / 060919; WO 98 / 23289; and WO 97 / 34631; and U.S. Patent Nos. 5,869,046, 6,121,022, 6,277,375, and 6,165,745, all of which are herein incorporated by reference in their entireties, for examples of mutations that will alter (e.g., decrease or increase) the half-life of an antibody in vivo. In certain embodiments, one, two or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into an IgG constant region, or FcRn-binding fragment thereof (preferably an Fc or hinge-Fc fragment) to decrease the half-life of the antibody in vivo. In other embodiments, one, two or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into an IgG constant region, or FcRn-binding fragment thereof (preferably an Fc or hinge-Fc fragment) to increase the half-life of the antibody in vivo. In a specific embodiment, the antibodies may have one or more amino acid mutations (e.g., substitutions) in the second constant (CH2) domain (residues 231-340 of human IgGi) and / or the third constant (CH3) domain (residues 341-447 of human IgGi), numberedaccording to the EU numbering system. In a specific embodiment, the constant region of the IgGi of antibody described herein comprises a methionine (M) to tyrosine (Y) substitution in position 252, a serine (S) to threonine (T) substitution in position 254, and a threonine (T) to glutamic acid (E) substitution in position 256, numbered according to the EU numbering system. See U.S. Patent No. 7,658,921, which is herein incorporated by reference in its entirety. This type of mutant IgG, referred to as “YTE mutant” has been shown to display fourfold increased half-life as compared to wild-type versions of the same antibody (see Dall’Acqua WF et al., (2006) J Biol Chem 281: 23514-24, which is herein incorporated by reference in its entirety). In certain embodiments, an antibody comprises an IgG constant region comprising one, two, three, or more amino acid substitutions of amino acid residues at positions 251-257, 285-290, 308-314, 385-389, and 428- 436, numbered according to the EU numbering system.

[0124] In certain embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into an Fc region (e.g., a CH2 domain (residues 231-340 of human IgGi)) and / or a CH3 domain (residues 341-447 of human IgGi, numbered according to the EU numbering system) and / or a hinge region (residues 216-230, numbered according to the EU numbering system) of an antibody described herein, to increase or decrease the affinity of the antibody for an Fc receptor (e.g., an activated Fc receptor) on the surface of an effector cell. Mutations in the Fc region of an antibody that decrease or increase the affinity of an antibody for an Fc receptor and techniques for introducing such mutations into the Fc receptor or fragment thereof are known to one of skill in the art. Examples of mutations in the Fc receptor of an antibody that can be made to alter the affinity of the antibody for an Fc receptor are described in, e.g., Smith P et al., (2012) PNAS 109: 6181-6186, U.S. Patent No. 6,737,056, and International Publication Nos. WO 02 / 060919; WO 98 / 23289; and WO 97 / 34631, all of which are herein incorporated by reference in their entireties.

[0125] In certain embodiments, the antibody comprises a heavy chain constant region that is a variant of a wild-type heavy chain constant region, wherein the variant heavy chain constant region binds to FcyRIIB with higher affinity than the wild-type heavy chain constant region binds to FcyRIIB. In certain embodiments, the variant heavy chain constant region is a variant human heavy chain constant region, e.g., a variant human IgGi, a variant human IgG2, or a variant human IgG4 heavy chain constant region. In certain embodiments, the variant human IgG heavy chain constant region comprises one or more of the following amino acid mutations, according to the EU numbering system: G236D, P238D, S239D, S267E, L328F, and L328E. In certain embodiments,the variant human IgG heavy chain constant region comprises a set of amino acid mutations selected from the group consisting of: S267E and L328F; P238D and L328E; P238D and one or more substitutions selected from the group consisting of E233D, G237D, H268D, P271G, and A330R; P238D, E233D, G237D, H268D, P271G, and A330R; G236D and S267E; S239D and S267E; V262E, S267E, and L328F; and V264E, S267E, and L328F, according to the EU numbering system. In certain embodiments, the FcyRIIB is expressed on a cell selected from the group consisting of macrophages, monocytes, B cells, dendritic cells, endothelial cells, and activated T cells.

[0126] In a further embodiment, one, two, or more amino acid substitutions are introduced into an IgG constant region Fc region to alter the effector function(s) of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 239, 243, 267, 292, 297, 300, 318, 320, 322, 328, 330, 332, and 396, numbered according to the EU numbering system, can be replaced with a different amino acid residue such that the antibody has an altered affinity for an effector ligand but retains the antigen-binding ability of the parent antibody. The effector ligand to which affinity is altered can be, for example, an Fc receptor or the Cl component of complement. This approach is described in further detail in U.S. Patent Nos. 5,624,821 and 5,648,260, each of which is herein incorporated by reference in its entirety. In certain embodiments, the deletion or inactivation (through point mutations or other means) of a constant region domain may reduce Fc receptor binding of the circulating antibody thereby increasing tumor localization. See, e.g., U.S. Patent Nos. 5,585,097 and 8,591,886, each of which is herein incorporated by reference in its entirety, for a description of mutations that delete or inactivate the constant region and thereby increase tumor localization. In certain embodiments, one or more amino acid substitutions may be introduced into the Fc region of an antibody described herein to remove potential glycosylation sites on the Fc region, which may reduce Fc receptor binding (see, e.g., Shields RL et al., (2001) J Biol Chem 276: 6591-604, which is herein incorporated by reference in its entirety). In various embodiments, one or more of the following mutations in the constant region of an antibody described herein may be made: an N297A substitution; an N297Q substitution; an L234A substitution; an L234F substitution; an L235A substitution; an L235F substitution; an L235V substitution; an L237A substitution; an S239D substitution; an E233P substitution; an L234V substitution; a C236 deletion; aP238A substitution; an F243L substitution; a D265A substitution; an S267E substitution; an L328F substitution; an R292P substitution; aY300L substitution; an A327Q substitution; a P329A substitution; an A330L substitution; an I332E substitution; or a P396L substitution, numbered according to the EU numbering system.

[0127] In certain embodiments, a mutation selected from the group consisting of D265A, P329A, and a combination thereof, numbered according to the EU numbering system, may be made in the constant region of an antibody described herein. In certain embodiments, a mutation selected from the group consisting of L235A, L237A, and a combination thereof, numbered according to the EU numbering system, may be made in the constant region of an antibody described herein. In certain embodiments, a mutation selected from the group consisting of S267E, L328F, and a combination thereof, numbered according to the EU numbering system, may be made in the constant region of an antibody described herein. In certain embodiments, a mutation selected from the group consisting of S239D, I332E, optionally A330L, and a combination thereof, numbered according to the EU numbering system, may be made in the constant region of an antibody described herein. In certain embodiments, a mutation selected from the group consisting of L235V, F243L, R292P, Y300L, P396L, and a combination thereof, numbered according to the EU numbering system, may be made in the constant region of an antibody described herein. In certain embodiments, a mutation selected from the group consisting of S267E, L328F, and a combination thereof, numbered according to the EU numbering system, may be made in the constant region of an antibody described herein.

[0128] In a specific embodiment, an antibody described herein comprises the constant region of an IgGi with an N297Q or N297A amino acid substitution, numbered according to the EU numbering system. In certain embodiments, an antibody described herein comprises the constant region of an IgGi with a mutation selected from the group consisting of D265A, P329A, and a combination thereof, numbered according to the EU numbering system. In another embodiment, an antibody described herein comprises the constant region of an IgGi with a mutation selected from the group consisting of L234A, L235A, and a combination thereof, numbered according to the EU numbering system. In another embodiment, an antibody described herein comprises the constant region of an IgGi with a mutation selected from the group consisting of L234F, L235F, N297A, and a combination thereof, numbered according to the EU numbering system. In certain embodiments, amino acid residues in the constant region of an antibody described herein in the positions corresponding to positions L234, L235, and D265 in a human IgGi heavy chain, numbered according to the EU numbering system, are not L, L, and D, respectively. This approachis described in detail in International Publication No. WO 14 / 108483, which is herein incorporated by reference in its entirety. In certain embodiments, the amino acids corresponding to positions L234, L235, and D265 in a human IgGi heavy chain are F, E, and A; or A, A, and A, respectively, numbered according to the EU numbering system.

[0129] In certain embodiments, one or more amino acids selected from amino acid residues 329, 331, and 322 in the constant region of an antibody described herein, numbered according to the EU numbering system, can be replaced with a different amino acid residue such that the antibody has altered Clq binding and / or reduced or abolished complement dependent cytotoxicity (CDC). This approach is described in further detail in U.S. Patent No. 6,194,551 (Idusogie et al.), which is herein incorporated by reference in its entirety. In certain embodiments, one or more amino acid residues within amino acid positions 231 to 238 in the N-terminal region of the CH2 domain of an antibody described herein are altered to thereby alter the ability of the antibody to fix complement, numbered according to the EU numbering system. This approach is described further in International Publication No. WO 94 / 29351, which is herein incorporated by reference in its entirety. In certain embodiments, the Fc region of an antibody described herein is modified to increase the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC) and / or to increase the affinity of the antibody for an Fey receptor by mutating one or more amino acids (e.g., introducing amino acid substitutions) at the following positions: 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290,292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322, 324, 326, 327, 328,329, 330, 331, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, 388, 389, 398, 414, 416,419, 430, 434, 435, 437, 438, or 439, numbered according to the EU numbering system. This approach is described further in International Publication No. WO 00 / 42072, which is herein incorporated by reference in its entirety.

[0130] In certain embodiments, an antibody described herein comprises a modified constant region of an IgGi, wherein the modification increases the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC). In certain embodiments, 0.1, 1, or 10 pg / ml of the antibody is capable of inducing cell death of at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% of NAPRT-expressing cells within 1, 2, or 3 hours, as assessed by methods described herein and / or known to a person of skill in the art. In certain embodiments, the modified constant region of an IgGi comprises S239D and I332E substitutions, numbered according to the EUnumbering system. In certain embodiments, the modified constant region of an IgGi comprises S239D, A330L, and I332E substitutions, numbered according to the EU numbering system. In certain embodiments, the modified constant region of an IgGi comprises L235V, F243L, R292P, Y300L, and P396L substitutions, numbered according to the EU numbering system. In certain embodiments, the antibody is capable of inducing cell death in effector T cells and Tregs, wherein the percentage of Tregs that undergo cell death is higher than the percentage of effector T cells that undergo cell death by at least 1.2 fold, 1.3 fold, 1.4 fold, 1.5 fold, 1.6 fold, 1.7 fold, 1.8 fold, 1.9 fold, 2 fold, 2.5 fold, 3 fold, 3.5 fold, 4 fold, 4.5 fold, or 5 fold.

[0131] In certain embodiments, an antibody described herein comprises the constant region of an IgG4antibody and the serine at amino acid residue 228 of the heavy chain, numbered according to the EU numbering system, is substituted for proline.

[0132] In certain embodiments, any of the constant region mutations or modifications described herein can be introduced into one or both heavy chain constant regions of an antibody described herein having two heavy chain constant regions.Polypeptides

[0133] In another aspect, provided herein are polypeptides comprising one or more sequences set forth in Tables 2-4, above. In certain embodiments, the polypeptide comprises the CDRH1, CDRH2, and / or CDRH3 of a VH amino acid sequence set forth in SEQ ID NO: 1, as determined by any of the methods discussed above. In certain embodiments, the polypeptide comprises a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NOs: 1. In certain embodiments, the polypeptide comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13; 15, 16, and 5; or 17, 18, and 19, respectively.

[0134] In certain embodiments, the polypeptide comprises the CDRL1, CDRL2, and / or CDRL3 of a VL amino acid sequence set forth in SEQ ID NO: 2, as determined by any of the methods discussed above. In certain embodiments, the polypeptide comprises a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NOs: 2. In certain embodiments, the polypeptide comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21, and 8, respectively.

[0135] In certain embodiments, the polypeptide comprises a VH comprising the CDRH1 , CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NO: 1; and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NO: 2. In certain embodiments, the polypeptide comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences of the VH and VL amino acid sequences set forth in SEQ ID NOs: 1 and 2, respectively.

[0136] In certain embodiments, a polypeptide provided herein comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 3, 4, 5, 6, 7, and 8; 9, 10, 5, 6, 7, and 8; 11, 12, 13, 14, LM, and 8; 11, 12, 13, 14, LMS, and 8; 15, 16, 5, 6, 7, and 8; or 17, 18, 19, 20, 21, and 8, respectively.Pharmaceutical Compositions

[0137] Provided herein are compositions comprising a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi), and optionally niacin, disclosed herein having the desired degree of purity in a physiologically acceptable carrier, excipient, or stabilizer (see, e.g., Remington’s Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA). Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, di saccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

[0138] In a specific embodiment, pharmaceutical compositions comprise a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi) disclosed herein, and optionally one or more additional prophylactic or therapeutic agents (e.g., niacin), in a pharmaceutically acceptablecarrier. In a specific embodiment, pharmaceutical compositions comprise a NAMPTi disclosed herein, and optionally one or more additional prophylactic or therapeutic agents (e.g., niacin), in a pharmaceutically acceptable carrier. In certain embodiments, the NAMPTi is the only active ingredient included in the pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises niacin and the NAMPTi. Pharmaceutical compositions described herein can be useful in decreasing or blocking NAMPTi (e.g., human NAMPTi) activity and treating a condition, such as cancer. In certain embodiments, the present disclosure relates to a pharmaceutical composition of the present disclosure comprising a NAMPTi for use as a medicament. In certain embodiments, the present disclosure relates to a pharmaceutical composition of the present disclosure comprising a NAMPTi, and optionally niacin, for use as a medicament. In another embodiment, the present disclosure relates to a pharmaceutical composition of the present disclosure for use in a method for the treatment of cancer.

[0139] Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharmaceutically acceptable substances. Examples of aqueous vehicles include sodium chloride injection, ringer’s injection, isotonic dextrose injection, sterile water injection, dextrose and lactated Ringer’s injection. Nonaqueous parenteral vehicles include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil, and peanut oil. Antimicrobial agents in bacteriostatic or fungistatic concentrations can be added to parenteral preparations packaged in multiple-dose containers which include phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxy benzoic acid esters, thimerosal, benzalkonium chloride, and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Emulsifying agents include Polysorbate 80 (TWEEN® 80). A sequestering or chelating agent of metal ions includes EDTA. Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol, and propylene glycol for water miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0140] A pharmaceutical composition can be formulated for any route of administration to a subject. Specific examples of routes of administration include intranasal, oral, pulmonary, transdermal, intradermal, and parenteral. Parenteral administration, characterized by either subcutaneous, intramuscular, or intravenous injection, is also contemplated herein. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. The injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. In addition, if desired, the pharmaceutical compositions to be administered can also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrins.

[0141] Preparations for parenteral administration of antibody include sterile solutions ready for injection, sterile dry soluble products, such as lyophilized powders, ready to be combined with a solvent just prior to use, including hypodermic tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with a vehicle just prior to use, and sterile emulsions. The solutions may be either aqueous or nonaqueous.

[0142] If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0143] Topical mixtures comprising an antibody are prepared as described for the local and systemic administration. The resulting mixture can be a solution, suspension, emulsions, or the like and can be formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, dermal patches, or any other formulations suitable for topical administration.

[0144] The NAMPTi disclosed herein can be formulated as an aerosol for topical application, such as by inhalation (see, e.g., U.S. Patent Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivery of a steroid useful for treatment of inflammatory diseases, particularly asthma and are herein incorporated by reference in their entireties). These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a microfine powder for insufflations, alone or in combination with an inert carriersuch as lactose. In such a case, the particles of the formulation will, in certain embodiments, have diameters of less than 50 microns, In certain embodiments less than 10 microns.

[0145] The NAMPTi disclosed herein can be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions and for application to the eye or for intraci sternal or intraspinal application. Topical administration is contemplated for transdermal delivery and also for administration to the eyes or mucosa, or for inhalation therapies. Nasal solutions of the antibody alone or in combination with other pharmaceutically acceptable excipients can also be administered.

[0146] Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those of skill in the art, and can be used to administer an antibody. For example, such patches are disclosed in U.S. Patent Nos. 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010715, 5,985,317, 5,983,134, 5,948,433, and 5,860,957, all of which are herein incorporated by reference in their entireties.

[0147] The NAMPTi disclosed herein and other compositions provided herein can also be formulated to be targeted to a particular tissue, receptor, or other area of the body of the subject to be treated. Many such targeting methods are well known to those of skill in the art. All such targeting methods are contemplated herein for use in the instant compositions. For non-limiting examples of targeting methods, see, e.g., U.S. Patent Nos. 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874, all of which are herein incorporated by reference in their entireties. In a specific embodiment, an antibody described herein is targeted to a tumor.

[0148] The compositions to be used for in vivo administration can be sterile. This is readily accomplished by filtration through, e.g., sterile filtration membranes.Methods of Use

[0149] The anti-NAPRT antibodies disclosed herein are particularly useful for the detection of NAPRT in a sample. In certain embodiments, NAPRT comprises the amino acid sequence of any one of SEQ ID NOs: 28-40.

[0150] In certain embodiments, the anti-NAPRT antibodies are useful in indicating levels of NAPRT in a sample. In certain embodiments, the anti-NAPRT antibodies are useful in indicatingthe likelihood of a subject with cancer to be effectively treated with a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi). In certain embodiments, the sample is obtained from a cancer or tumor from the subject. In certain embodiments, the subject is a human subject. In certain embodiments, the cancer has an increased likelihood to be effectively treated with an NAMPTi if the NAPRT levels from the sample are lower than in a control sample. In certain embodiments, the cancer may be effectively treated with an NAMPTi if no NAPRT is detected in the sample.

[0151] In certain embodiments, the sample comprises cancer cells. In certain embodiments the cancer cells are solid tumor cells, such as thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer cells.

[0152] In certain embodiments, the cancer that has increased likelihood to be effectively treated with an NAMPTi is thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer. In certain embodiments, the cancer is treated with the administration of niacin in addition to the NAMPTi.

[0153] In another aspect, provided herein is a treating cancer comprising administering a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi) to a subject in need thereof, wherein a sample from the cancer has lower NAPRT levels than a control sample. In certain embodiments, the method further comprises administering niacin to the subject.

[0154] In another aspect, provided herein is a method of diagnosing a subject with a cancer with increased likelihood to be effectively treated with a NAMPTi comprising administering an antibody that specifically binds to NAPRT to a sample from the subject and detecting specific binding of the antibody to NAPRT, wherein if there is less binding to NAPRT in the sample from the subject than in a control sample, the cancer in the subject has an increased likelihood to be effectively treated with the NAMPTi.

[0155] In another aspect, the anti-NAPRT antibodies disclosed herein are useful for the diagnosis of sepsis. In certain embodiments, the anti-NAPRT antibodies are useful in indicating the likelihood of subject to have sepsis. In certain embodiments, a subject has an increase likelihood to have sepsis if the detected level of NAPRT in a sample from the subject is higher than the detected level in a control sample.Polynucleotides, Vectors, Recombinant Host Cells, and Methods of Producing Antibodies

[0156] In another aspect, provided herein are polynucleotides comprising a nucleotide sequence encoding an antibody, or a portion thereof, described herein or a fragment thereof (e.g, a VL and / or VH; and a light chain and / or heavy chain) that specifically binds to an NAPRT (e.g., human NAPRT) antigen, and vectors, e.g., vectors comprising such polynucleotides for recombinant expression in host cells (e.g., E. coli and mammalian cells). Provided herein are polynucleotides comprising nucleotide sequences encoding a heavy and / or light chain of any of the antibodies provided herein, as well as vectors comprising such polynucleotide sequences, e.g., expression vectors for their efficient expression in host cells, e.g., mammalian cells.

[0157] As used herein, an “isolated” polynucleotide or nucleic acid molecule is one which is separated from other nucleic acid molecules which are present in the natural source (e.g., in a mouse or a human) of the nucleic acid molecule. Moreover, an “isolated” nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. For example, the language “substantially free” includes preparations of polynucleotide or nucleic acid molecules having less than about 15%, 10%, 5%, 2%, 1%, 0.5%, or 0.1% (in particular less than about 10%) of other material, e.g., cellular material, culture medium, other nucleic acid molecules, chemical precursors, and / or other chemicals. In a specific embodiment, a nucleic acid molecule(s) encoding an antibody described herein is isolated or purified.

[0158] In particular aspects, provided herein are polynucleotides comprising nucleotide sequences encoding antibodies, which specifically bind to an NAPRT (e.g., human NAPRT) polypeptide and comprises an amino acid sequence as described herein, as well as antibodies which compete with such antibodies for binding to an NAPRT (e.g., human NAPRT) polypeptide (e.g., in a dose-dependent manner), or which binds to the same epitope as that of such antibodies.

[0159] In certain aspects, provided herein are polynucleotides comprising a nucleotide sequence encoding the light chain or heavy chain of antibody described herein. The polynucleotides can comprise nucleotide sequences encoding a light chain comprising the VL FRs and CDRs of antibodies described herein (see, e.g., Tables 2 and 4) or nucleotide sequences encoding a heavy chain comprising the VH FRs and CDRs of antibodies described herein (see, e.g., Tables 2 and 3). In certain embodiments, a polynucleotide encodes a VH, VL, heavy chain, and / or light chain of an antibody described herein. In another embodiment, a polynucleotide encodes the first VH and the first VL of an antibody described herein. In another embodiment, a polynucleotide encodes the second VH and the second VL of an antibody described herein. In another embodiment, a polynucleotide encodes the first heavy chain and the first light chain of an antibody described herein. In another embodiment, a polynucleotide encodes the second heavy chain and the second light chain of an antibody described herein. In another embodiment, a polynucleotide encodes the VH and / or the VL, or the heavy chain and / or the light chain, of an antibody described herein.

[0160] Also provided herein are polynucleotides encoding an anti-NAPRT antibody that are optimized, e.g., by codon / RNA optimization, replacement with heterologous signal sequences, and elimination of mRNA instability elements. Methods to generate optimized nucleic acids encoding an anti-NAPRT antibody or a fragment thereof (e.g., light chain, heavy chain, VH domain, or VL domain) for recombinant expression by introducing codon changes and / or eliminating inhibitory regions in the mRNA can be carried out by adapting the optimization methods described in, e.g., U.S. Patent Nos. 5,965,726; 6,174,666; 6,291,664; 6,414,132; and 6,794,498, accordingly, all of which are herein incorporated by reference in their entireties. For example, potential splice sites and instability elements (e.g., A / T or A / U rich elements) within the RNA can be mutated without altering the amino acids encoded by the nucleic acid sequences to increase stability of the RNA for recombinant expression. The alterations utilize the degeneracy of the genetic code, e.g., using an alternative codon for an identical amino acid. In certain embodiments, it can be desirable to alter one or more codons to encode a conservative mutation, e.g., a similar amino acid with similar chemical structure and properties and / or function as the original amino acid. Such methods can increase expression of an anti-NAPRT antibody or fragment thereof by at least 1 fold, 2 fold, 3 fold, 4 fold, 5 fold, 10 fold, 20 fold, 30 fold, 40 fold, 50 fold, 60 fold, 70 fold, 80 fold, 90 fold, or100 fold or more relative to the expression of an anti-NAPRT antibody encoded by polynucleotides that have not been optimized.

[0161] In certain embodiments, an optimized polynucleotide sequence encoding an anti- NAPRT antibody described herein or a fragment thereof (e.g., VL domain and / or VH domain) can hybridize to an antisense (e.g., complementary) polynucleotide of an unoptimized polynucleotide sequence encoding an anti-NAPRT antibody described herein or a fragment thereof e.g., VL domain and / or VH domain). In specific embodiments, an optimized nucleotide sequence encoding an anti-NAPRT antibody described herein or a fragment hybridizes under high stringency conditions to antisense polynucleotide of an unoptimized polynucleotide sequence encoding an anti-NAPRT antibody described herein or a fragment thereof. In a specific embodiment, an optimized nucleotide sequence encoding an anti-NAPRT antibody described herein or a fragment thereof hybridizes under high stringency, intermediate or lower stringency hybridization conditions to an antisense polynucleotide of an unoptimized nucleotide sequence encoding an anti- NAPRT antibody described herein or a fragment thereof. Information regarding hybridization conditions has been described, see, e.g., U.S. Patent Application Publication No. US 2005 / 0048549 (e.g., paragraphs 72-73), which is herein incorporated by reference in its entirety.

[0162] The polynucleotides can be obtained, and the nucleotide sequence of the polynucleotides determined, by any method known in the art. Nucleotide sequences encoding antibodies described herein, e.g., antibodies described in Tables 2-4, and modified versions of these antibodies can be determined using methods well known in the art, i.e., nucleotide codons known to encode particular amino acids are assembled in such a way to generate a nucleic acid that encodes the antibody. Such a polynucleotide encoding the antibody can be assembled from chemically synthesized oligonucleotides (e.g., as described in Kutmeier G et al., (1994), BioTechniques 17: 242-6, herein incorporated by reference in its entirety), which, briefly, involves the synthesis of overlapping oligonucleotides containing portions of the sequence encoding the antibody, annealing and ligating of those oligonucleotides, and then amplification of the ligated oligonucleotides by PCR.

[0163] Alternatively, a polynucleotide encoding an antigen-binding region of an antibody described herein can be generated from nucleic acid from a suitable source (e.g., a hybridoma) using methods well known in the art (e.g., PCR and other molecular cloning methods). For example, PCR amplification using synthetic primers hybridizable to the 3’ and 5’ ends of a knownsequence can be performed using genomic DNA obtained from hybridoma cells producing the antibody of interest. Such PCR amplification methods can be used to obtain nucleic acids comprising the sequence encoding the light chain and / or heavy chain of an antibody. Such PCR amplification methods can be used to obtain nucleic acids comprising the sequence encoding the variable light chain region and / or the variable heavy chain region of an antibody. The amplified nucleic acids can be cloned into vectors for expression in host cells and for further cloning.

[0164] If a clone containing a nucleic acid encoding a particular antigen-binding region or antibody is not available, but the sequence of the antigen-binding region or antibody molecule is known, a nucleic acid encoding the immunoglobulin can be chemically synthesized or obtained from a suitable source (e.g., an antibody cDNA library or a cDNA library generated from, or nucleic acid, preferably poly A+ RNA, isolated from, any tissue or cells expressing the antibody, such as hybridoma cells selected to express an antibody described herein) by PCR amplification using synthetic primers hybridizable to the 3’ and 5’ ends of the sequence, or by cloning using an oligonucleotide probe specific for the particular gene sequence to identify, e.g., a cDNA clone from a cDNA library that encodes the antibody. Amplified nucleic acids generated by PCR can then be cloned into replicable cloning vectors using any method well known in the art.

[0165] DNA encoding anti-NAPRT (e.g., human NAPRT) antibodies described herein can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the anti-NAPRT (e.g., human NAPRT antibodies). Hybridoma cells can serve as a source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells (e.g., CHO cells from the CHO GS System™ (Lonza)), or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of anti-NAPRT antibodies in the recombinant host cells.

[0166] To generate whole antibodies or antigen-binding regions, PCR primers including VH or VL nucleotide sequences, a restriction site, and a flanking sequence to protect the restriction site can be used to amplify the VH or VL sequences in scFv clones. Utilizing cloning techniques known to those of skill in the art, the PCR amplified VH domains can be cloned into vectors expressing a heavy chain constant region, e.g., the human gamma 1 or human gamma 4 constant region, and the PCR amplified VL domains can be cloned into vectors expressing a light chainconstant region, e.g., human kappa or lambda constant regions. In certain embodiments, the vectors for expressing the VH or VL domains comprise an EF-la promoter, a secretion signal, a cloning site for the variable region, constant regions, and a selection marker such as neomycin. The VH and VL domains can also be cloned into one vector expressing the necessary constant regions. The heavy chain conversion vectors and light chain conversion vectors are then co-transfected into cell lines to generate stable or transient cell lines that express full-length antibodies, e.g., IgG, using techniques known to those of skill in the art.

[0167] The DNA also can be modified, for example, by substituting the coding sequence for human heavy and light chain constant regions in place of the murine sequences, or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a nonimmunoglobulin polypeptide.

[0168] Also provided are polynucleotides that hybridize under high stringency, intermediate or lower stringency hybridization conditions to polynucleotides that encode an antibody described herein. In specific embodiments, polynucleotides described herein hybridize under high stringency, intermediate or lower stringency hybridization conditions to polynucleotides encoding a VH domain and / or VL domain provided herein.

[0169] Hybridization conditions have been described in the art and are known to one of skill in the art. For example, hybridization under stringent conditions can involve hybridization to filterbound DNA in 6x sodium chloride / sodium citrate (SSC) at about 45°C followed by one or more washes in 0.2xSSC / 0.1% SDS at about 50-65°C; hybridization under highly stringent conditions can involve hybridization to filter-bound nucleic acid in 6xSSC at about 45°C followed by one or more washes in 0.1xSSC / 0.2% SDS at about 68°C. Hybridization under other stringent hybridization conditions are known to those of skill in the art and have been described, see, e.g., Ausubel FM et al., eds., (1989) Current Protocols in Molecular Biology, Vol. I, Green Publishing Associates, Inc. and John Wiley & Sons, Inc., New York at pages 6.3.1-6.3.6 and 2.10.3, which is herein incorporated by reference in its entirety.

[0170] In certain aspects, provided herein are cells (e.g., host cells) expressing (e.g., recombinantly) antibodies described herein which specifically bind to NAPRT (e.g., human NAPRT), and related polynucleotides and expression vectors. Provided herein are vectors (e.g., expression vectors) comprising polynucleotides comprising nucleotide sequences encoding anti- NAPRT antibodies or a fragment for recombinant expression in host cells, preferably inmammalian cells (e.g., CHO cells). Also provided herein are host cells comprising such vectors for recombinantly expressing anti-NAPRT antibodies described herein (e.g., human or humanized antibody). In a particular aspect, provided herein are methods for producing an antibody described herein, comprising expressing the antibody from a host cell.

[0171] Recombinant expression of an antibody described herein (e.g., a full-length antigenbinding region or antibody or heavy and / or light chain of an antibody described herein) that specifically binds to NAPRT (e.g., human NAPRT) generally involves construction of an expression vector containing a polynucleotide that encodes the antibody. Once a polynucleotide encoding an antibody molecule, heavy and / or light chain of an antibody, or a fragment thereof (e.g., heavy and / or light chain variable regions) described herein has been obtained, the vector for the production of the antibody molecule can be produced by recombinant DNA technology using techniques well known in the art. Thus, methods for preparing a protein by expressing a polynucleotide containing an antibody or antibody fragment (e.g., light chain or heavy chain) encoding nucleotide sequence are described herein. Methods which are well known to those skilled in the art can be used to construct expression vectors containing an antibody or antibody fragment (e.g., light chain or heavy chain) coding sequences and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Also provided are replicable vectors comprising a nucleotide sequence encoding containing an antibody molecule described herein, a heavy or light chain of an antibody, a heavy or light chain variable region of an antibody or a fragment thereof, or a heavy or light chain CDR, operably linked to a promoter. Such vectors can, for example, include the nucleotide sequence encoding the constant region of the antibody molecule (see, e.g., International Publication Nos. WO 86 / 05807 and WO 89 / 01036; and U.S. Patent No. 5,122,464, which are herein incorporated by reference in their entireties) and variable regions of the antibody can be cloned into such a vector for expression of the entire heavy, the entire light chain, or both the entire heavy and light chains.

[0172] In certain embodiments, a vector comprises a polynucleotide encoding a VH, VL, heavy chain, and / or light chain of an antibody described herein. In another embodiment, a vector comprises a polynucleotide encoding the VH and the VL of an antibody described herein. In another embodiment, a vector comprises a polynucleotide encoding the heavy chain and the light chain of an antibody described herein.

[0173] An expression vector can be transferred to a cell (e.g., host cell) by conventional techniques and the resulting cells can then be cultured by conventional techniques to produce cells containing an antibody described herein or a fragment thereof. Thus, provided herein are host cells containing a polynucleotide encoding containing an antibody described herein or fragments thereof, or a heavy or light chain thereof, or fragment thereof, or a single-chain antibody described herein, operably linked to a promoter for expression of such sequences in the host cell.

[0174] In certain embodiments, a host cell comprises a polynucleotide encoding the VH and VL of an antibody described herein. In another embodiment, a host cell comprises a vector comprising a polynucleotide encoding the VH and VL of an antibody described herein. In another embodiment, a host cell comprises a first polynucleotide encoding the VH of an antibody described herein, and a second polynucleotide encoding the VL of an antibody described herein. In another embodiment, a host cell comprises a first vector comprising a first polynucleotide encoding the VH of an antibody described herein, and a second vector comprising a second polynucleotide encoding the VL of an antibody described herein.

[0175] In specific embodiments, a heavy chain / heavy chain variable region expressed by a first cell is associated with a light chain / light chain variable region of a second cell to form an anti- NAPRT (e.g., human NAPRT) antibody described herein. In certain embodiments, provided herein is a population of host cells comprising such first host cell and such second host cell.

[0176] In certain embodiments, provided herein is a population of vectors comprising a first vector comprising a polynucleotide encoding a light chain / light chain variable region of an anti- NAPRT (e.g., human NAPRT) antibody described herein, and a second vector comprising a polynucleotide encoding a heavy chain / heavy chain variable region of an anti -NAPRT (e.g., human NAPRT) antibody described herein.

[0177] A variety of host-expression vector systems can be utilized to express antibody molecules described herein (see, e.g., U.S. Patent No. 5,807,715, which is herein incorporated by reference in its entirety). Such host-expression systems represent vehicles by which the coding sequences of interest can be produced and subsequently purified, but also represent cells which can, when transformed or transfected with the appropriate nucleotide coding sequences, express an antibody molecule described herein in situ. These include but are not limited to microorganisms such as bacteria (e.g., E. coli and B. subtilis') transformed with, e.g., recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing antibody coding sequences;yeast (e.g., Saccharomyces and Pichia) transformed with, e.g., recombinant yeast expression vectors containing antibody coding sequences; insect cell systems infected with, e.g., recombinant virus expression vectors (e.g., baculovirus) containing antibody coding sequences; plant cell systems (e.g., green algae such as Chlamydomonas reinhardtii) infected with, e.g., recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with, e.g., recombinant plasmid expression vectors (e.g., Ti plasmid) containing antibody coding sequences; or mammalian cell systems (e.g., COS (e.g., COS1 or COS), CHO, BHK, MDCK, HEK 293, NSO, PER.C6, VERO, CRL7O3O, HsS78Bst, HeLa, and NIH 3T3, HEK-293T, HepG2, SP210, Rl. l, B-W, L-M, BSC1, BSC40, YB / 20 and BMTIO cells) harboring, e.g., recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter). In a specific embodiment, cells for expressing antibodies described herein are Chinese hamster ovary (CHO) cells, for example CHO cells from the CHO GS System™ (Lonza). In certain embodiments, the heavy chain and / or light chain of an antibody produced by a CHO cell may have an N-terminal glutamine or glutamate residue replaced by pyroglutamate. In certain embodiments, cells for expressing antibodies described herein are human cells, e.g., human cell lines. In a specific embodiment, a mammalian expression vector is pOptiVEC™ or pcDNA3.3. In certain embodiments, bacterial cells such as Escherichia coli, or eukaryotic cells (e.g., mammalian cells), especially for the expression of whole recombinant antibody molecule, are used for the expression of a recombinant antibody molecule. For example, mammalian cells such as CHO cells, in conjunction with a vector such as the major intermediate early gene promoter element from human cytomegalovirus, are an effective expression system for antibodies (Foecking MK & Hofstetter H (1986) Gene 45: 101-5; and Cockett MI et al., (1990) Biotechnology 8(7): 662-7, each of which is herein incorporated by reference in its entirety). In certain embodiments, antibodies described herein are produced by CHO cells or NSO cells. In a specific embodiment, the expression of nucleotide sequences encoding antibodies described herein which specifically bind to NAPRT (e.g., human NAPRT) is regulated by a constitutive promoter, inducible promoter, or tissue specific promoter.

[0178] In bacterial systems, a number of expression vectors can be advantageously selected depending upon the use intended for the antibody molecule being expressed. For example, when a large quantity of such an antibody is to be produced, for the generation of pharmaceuticalcompositions of an antibody molecule, vectors which direct the expression of high levels of fusion protein products that are readily purified can be desirable. Such vectors include, but are not limited to, the E. colt expression vector pUR278 (Ruether U & Mueller-Hill B (1983) EMBO J 2: 1791- 1794), in which the coding sequence can be ligated individually into the vector in frame with the lac Z coding region so that a fusion protein is produced; pIN vectors (Inouye S & Inouye M (1985) Nuc Acids Res 13: 3101-3109; Van Heeke G & Schuster SM (1989) J Biol Chem 24: 5503-5509); and the like, all of which are herein incorporated by reference in their entireties. For example, pGEX vectors can also be used to express foreign polypeptides as fusion proteins with glutathione 5-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption and binding to matrix glutathione agarose beads followed by elution in the presence of free glutathione. The pGEX vectors are designed to include thrombin or factor Xa protease cleavage sites so that the cloned target gene product can be released from the GST moiety.

[0179] In an insect system, Autographa californica nuclear polyhedrosis virus (AcNPV), for example, can be used as a vector to express foreign genes. The virus grows in Spodoptera frugiperda cells. The coding sequence can be cloned individually into non-essential regions (for example the polyhedrin gene) of the virus and placed under control of an AcNPV promoter (for example the polyhedrin promoter).

[0180] In mammalian host cells, a number of viral-based expression systems can be utilized. In cases where an adenovirus is used as an expression vector, the coding sequence of interest can be ligated to an adenovirus transcription / translation control complex, e.g., the late promoter and tripartite leader sequence. This chimeric gene can then be inserted in the adenovirus genome by in vitro or in vivo recombination. Insertion in a non-essential region of the viral genome (e.g., region El or E3) will result in a recombinant virus that is viable and capable of expressing the molecule in infected hosts (see, e.g., Logan J & Shenk T (1984) PNAS 81(12): 3655-9, which is herein incorporated by reference in its entirety). Specific initiation signals can also be required for efficient translation of inserted coding sequences. These signals include the ATG initiation codon and adjacent sequences. Furthermore, the initiation codon must be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert. These exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic. The efficiency of expression can be enhanced by the inclusion of appropriatetranscription enhancer elements, transcription terminators, etc. (see, e.g., Bitter G et al., (1987) Methods Enzymol. 153: 516-544, which is herein incorporated by reference in its entirety).

[0181] In addition, a host cell strain can be chosen which modulates the expression of the inserted sequences or modifies and processes the gene product in the specific fashion desired. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products can be important for the function of the protein. Different host cells have characteristic and specific mechanisms for the post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the foreign protein expressed. To this end, eukaryotic host cells which possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product can be used. Such mammalian host cells include but are not limited to CHO, VERO, BHK, Hela, MDCK, HEK 293, NIH 3T3, W138, BT483, Hs578T, HTB2, BT2O and T47D, NSO (a murine myeloma cell line that does not endogenously produce any immunoglobulin chains), CRL7O3O, COS (e.g., COS1 or COS), PER.C6, VERO, HsS78Bst, HEK-293T, HepG2, SP210, Rl.l, B-W, L-M, BSC1, BSC40, YB / 20, BMT10, and HsS78Bst cells. In certain embodiments, anti-NAPRT (e.g., human NAPRT) antibodies described herein are produced in mammalian cells, such as CHO cells.

[0182] In a specific embodiment, the antibodies described herein have reduced fucose content or no fucose content. Such antibodies can be produced using techniques known one skilled in the art. For example, the antibodies can be expressed in cells deficient or lacking the ability to fucosylate. In a specific example, cell lines with a knockout of both alleles of al, 6- fucosyltransferase can be used to produce antibodies with reduced fucose content. The Potelligent® system (Lonza) is an example of such a system that can be used to produce antibodies with reduced fucose content.

[0183] For long-term, high-yield production of recombinant proteins, stable expression cells can be generated. For example, cell lines which stably express an anti-NAPRT (e.g., human NAPRT) antibody described herein can be engineered. In specific embodiments, a cell provided herein stably expresses a light chain / light chain variable region and a heavy chain / heavy chain variable region which associate to form an antigen-binding region or an antibody described herein.

[0184] In certain aspects, rather than using expression vectors which contain viral origins of replication, host cells can be transformed with DNA controlled by appropriate expression controlelements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of the foreign DNA / polynucleotide, engineered cells can be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci which in turn can be cloned and expanded into cell lines. This method can advantageously be used to engineer cell lines which express an anti-NAPRT (e.g., human NAPRT) described herein or a fragment thereof. Such engineered cell lines can be particularly useful in screening and evaluation of compositions that interact directly or indirectly with the antibody molecule.

[0185] A number of selection systems can be used, including but not limited to the herpes simplex virus thymidine kinase (Wigler Me / al., (1977) Cell 11(1): 223-32), hypoxanthineguanine phosphoribosyltransferase (Szybalska EH & Szybalski W (1962) PNAS 48(12): 2026-2034), and adenine phosphoribosyltransferase (Lowy I et aL, (1980) Cell 22(3): 817-23) genes in tk-, hgprt- or aprt-cells, respectively, all of which are herein incorporated by reference in their entireties. Also, antimetabolite resistance can be used as the basis of selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler M et al., (1980) PNAS 77(6): 3567-70; O’Hare K et a , (1981) PNAS 78: 1527-31); gpt, which confers resistance to mycophenolic acid (Mulligan RC & Berg P (1981) PNAS 78(4): 2072-6); neo, which confers resistance to the aminoglycoside G- 418 (Wu GY & Wu CH (1991) Biotherapy 3: 87-95; Tolstoshev P (1993) Ann Rev Pharmacol Toxicol 32: 573-596; Mulligan RC (1993) Science 260: 926-932; and Morgan RA & Anderson WF (1993) Ann Rev Biochem 62: 191-217; Nabel GJ & Feigner PL (1993) Trends Biotechnol 11(5): 211-5); and hygro, which confers resistance to hygromycin (Santerre RF etal., (1984) Gene 30(1-3): 147-56), all of which are herein incorporated by reference in their entireties. Methods commonly known in the art of recombinant DNA technology can be routinely applied to select the desired recombinant clone and such methods are described, for example, in Ausubel FM et al., (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993); Kriegler M, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990); and in Chapters 12 and 13, Dracopoli NC et aL, (eds.), Current Protocols in Human Genetics, John Wiley & Sons, NY (1994); Colbere-Garapin F et aL, (1981) J Mol Biol 150: 1-14, all of which are herein incorporated by reference in their entireties.

[0186] The expression levels of an antibody molecule can be increased by vector amplification (for a review, see Bebbington CR & Hentschel CCG, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol. 3 (Academic Press, New York, 1987), which is herein incorporated by reference in its entirety). When a marker in the vector system is amplifiable, an increase in the level of inhibitor present in culture of the host cell will increase the number of copies of the marker gene. Since the amplified region is associated with the gene of interest, production of the protein will also increase (Crouse GF el al., (1983) Mol Cell Biol 3 : 257-66, which is herein incorporated by reference in its entirety).

[0187] The host cell can be co-transfected with two or more expression vectors described herein, the first vector encoding a heavy chain derived polypeptide and the second vector encoding a light chain derived polypeptide. The two vectors can contain identical selectable markers which enable equal expression of heavy and light chain polypeptides. The host cells can be co-transfected with different amounts of the two or more expression vectors. For example, host cells can be transfected with any one of the following ratios of a first expression vector and a second expression vector: about 1: 1, 1 :2, 1 :3, 1 :4, 1 :5, 1 :6, 1 :7, 1 :8, 1 :9, 1 : 10, 1 :12, 1 :15, 1 :20, 1:25, 1 :30, 1 :35, 1 :40, 1 :45, or 1:50.

[0188] Alternatively, a single vector can be used which encodes, and is capable of expressing, both heavy and light chain polypeptides. In such situations, the light chain should be placed before the heavy chain to avoid an excess of toxic free heavy chain (Proudfoot NJ (1986) Nature 322: 562-565; and Kohler G (1980) PNAS 77: 2197-2199, each of which is herein incorporated by reference in its entirety). The coding sequences for the heavy and light chains can comprise cDNA or genomic DNA. The expression vector can be monocistronic or multicistronic. A multi ci stronic nucleic acid construct can encode 2, 3, 4, 5, 6, 7, 8, 9, 10, or more genes / nucleotide sequences, or in the range of 2-5, 5-10, or 10-20 genes / nucleotide sequences. For example, a bicistronic nucleic acid construct can comprise, in the following order, a promoter, a first gene (e.g., heavy chain of an antibody described herein), and a second gene and (e.g., light chain of an antibody described herein). In such an expression vector, the transcription of both genes can be driven by the promoter, whereas the translation of the mRNA from the first gene can be by a cap-dependent scanning mechanism and the translation of the mRNA from the second gene can be by a cap-independent mechanism, e.g., by an IRES.

[0189] Once an antibody molecule described herein has been produced by recombinant expression, it can be purified by any method known in the art for purification of an immunoglobulin molecule, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for the purification of proteins. Further, the antibodies described herein can be fused to heterologous polypeptide sequences described herein or otherwise known in the art to facilitate purification.

[0190] In specific embodiments, an antibody described herein is isolated or purified. In certain embodiments, an isolated antibody is one that is substantially free of other antibodies with different antigenic specificities than the isolated antibody. For example, in certain embodiments, a preparation of an antibody described herein is substantially free of cellular material and / or chemical precursors. The language “substantially free of cellular material” includes preparations of an antibody in which the antibody is separated from cellular components of the cells from which it is isolated or recombinantly produced. Thus, an antibody that is substantially free of cellular material includes preparations of antibody having less than about 30%, 20%, 10%, 5%, 2%, 1%, 0.5%, or 0.1% (by dry weight) of heterologous protein (also referred to herein as a “contaminating protein”) and / or variants of an antibody, for example, different post-translational modified forms of an antibody or other different versions of an antibody (e.g., antibody fragments). When the antibody is recombinantly produced, it is also generally substantially free of culture medium, i.e., culture medium represents less than about 20%, 10%, 2%, 1%, 0.5%, or 0.1% of the volume of the protein preparation. When the antibody is produced by chemical synthesis, it is generally substantially free of chemical precursors or other chemicals, i.e., it is separated from chemical precursors or other chemicals which are involved in the synthesis of the protein. Accordingly, such preparations of the antibody have less than about 30%, 20%, 10%, or 5% (by dry weight) of chemical precursors or compounds other than the antibody of interest. In a specific embodiment, antibodies described herein are isolated or purified.

[0191] Anti-NAPRT (e.g., human NAPRT) antibodies or fragments thereof can be produced by any method known in the art for the synthesis of proteins or antibodies, for example, by chemical synthesis or by recombinant expression techniques. The methods described herein employ, unless otherwise indicated, conventional techniques in molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotidesynthesis and modification, nucleic acid hybridization, and related fields within the skill of the art. These techniques are described, for example, in the references cited herein and are fully explained in the literature. See, e.g., Maniatis T etal., (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Sambrook J et al., (1989), Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press; Sambrook J et al., (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel FM et al., Current Protocols in Molecular Biology, John Wiley & Sons (1987 and annual updates); Current Protocols in Immunology, John Wiley & Sons (1987 and annual updates) Gait (ed.) (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press; Eckstein (ed.) (1991) Oligonucleotides and Analogues: A Practical Approach, IRL Press; Birren B et al., (eds.) (1999) Genome Analysis: A Laboratory Manual, Cold Spring Harbor Laboratory Press, all of which are herein incorporated by reference in their entireties.

[0192] In a specific embodiment, an antibody described herein is prepared, expressed, created, or isolated by any means that involves creation, e.g., via synthesis, genetic engineering of DNA sequences. In certain embodiments, such an antibody comprises sequences (e.g., DNA sequences or amino acid sequences) that do not naturally exist within the antibody germline repertoire of an animal or mammal (e.g., human) in vivo.

[0193] In one aspect, provided herein is a method of making an anti-NAPRT (e.g., human NAPRT) antibody comprising culturing a cell or host cell described herein. In certain embodiments, the method is performed in vitro. In a certain aspect, provided herein is a method of making an anti-NAPRT (e.g., human NAPRT) antibody comprising expressing (e.g., recombinantly expressing) the antibody using a cell or host cell described herein (e.g., a cell or a host cell comprising polynucleotides encoding an antibody described herein). In certain embodiments, the cell is an isolated cell. In certain embodiments, the exogenous polynucleotides have been introduced into the cell. In certain embodiments, the method further comprises the step of purifying the antibody obtained from the cell or host cell.

[0194] In certain embodiments, an antibody is produced by expressing in a cell a polynucleotide encoding the VH and VL of an antibody described herein under suitable conditions so that the polynucleotide is expressed and the antibody is produced. In another embodiment, an antibody is produced by expressing in a cell a polynucleotide encoding the heavy chain and light chain of an antibody described herein under suitable conditions so that the polynucleotide isexpressed and the antibody is produced. Tn certain embodiments, an antibody is produced by expressing in a cell a first polynucleotide encoding the VH of an antibody described herein, and a second polynucleotide encoding the VL of an antibody described herein, under suitable conditions so that the polynucleotides are expressed and the antibody is produced. In certain embodiments, an antibody is produced by expressing in a cell a first polynucleotide encoding the heavy chain of an antibody described herein, and a second polynucleotide encoding the light chain of an antibody described herein, under suitable conditions so that the polynucleotides are expressed and the antibody is produced.

[0195] Methods for producing polyclonal antibodies are known in the art (see, e.g., Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel FM et al., eds., John Wiley and Sons, New York, which is herein incorporated by reference in its entirety).

[0196] Monoclonal antibodies can be prepared using a wide variety of techniques known in the art, including the use of hybridoma, recombinant, and phage display technologies, or a combination thereof. For example, monoclonal antibodies can be produced using hybridoma techniques, including those known in the art and taught, for example, in Harlow E & Lane D, Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling GJ etal., in: Monoclonal Antibodies and T-Cell Hybridomas 563 681 (Elsevier, N.Y., 1981), each of which is herein incorporated by reference in its entirety. The term “monoclonal antibody” as used herein is not limited to antibodies produced through hybridoma technology. For example, monoclonal antibodies can be produced recombinantly from host cells exogenously expressing an antibody described herein or a fragment thereof, for example, light chain and / or heavy chain of such antibody.

[0197] In specific embodiments, a “monoclonal antibody,” as used herein, is an antibody produced by a single cell (e.g., hybridoma or host cell producing a recombinant antibody), wherein the antibody specifically binds to NAPRT (e.g., human NAPRT) as determined, e.g., by ELISA or other antigen-binding or competitive binding assay known in the art or in the examples provided herein. In certain embodiments, a monoclonal antibody can be a chimeric antibody or a humanized antibody. In certain embodiments, a monoclonal antibody is a monovalent antibody or multivalent (e.g., bivalent) antibody. In certain embodiments, a monoclonal antibody is a monospecific or multispecific antibody (e.g., bispecific antibody). Monoclonal antibodies described herein can, for example, be made by the hybridoma method as described in Kohler G& Milstein C (1975) Nature256: 495, which is herein incorporated by reference in its entirety, or can, e.g., be isolated from phage libraries using the techniques as described herein, for example. Other methods for the preparation of clonal cell lines and of monoclonal antibodies expressed thereby are well known in the art (see, e.g., Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel FM et al., supra).

[0198] As used herein, an antibody binds to an antigen multivalently e.g., bivalently) when the antibody comprises at least two (e.g., two or more) monovalent binding regions, each monovalent binding region capable of binding to an epitope on the antigen. Each monovalent binding region can bind to the same or different epitopes on the antigen.

[0199] Methods for producing and screening for specific antibodies using hybridoma technology are routine and well known in the art. For example, in the hybridoma method, a mouse or other appropriate host animal, such as a sheep, goat, rabbit, rat, hamster, or macaque monkey, is immunized to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein (e.g., NAPRT (e.g., human NAPRT)) used for immunization. Alternatively, lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding JW (Ed.), Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986), herein incorporated by reference in its entirety). Additionally, a RIMMS (repetitive immunization multiple sites) technique can be used to immunize an animal (Kilpatrick KE et al., (1997) Hybridoma 16:381-9, herein incorporated by reference in its entirety).

[0200] In certain embodiments, mice (or other animals, such as rats, monkeys, donkeys, pigs, sheep, hamster, or dogs) can be immunized with an antigen (e.g., NAPRT (e.g., human NAPRT)), and once an immune response is detected, e.g., antibodies specific for the antigen are detected in the mouse serum, the mouse spleen is harvested and splenocytes isolated. The splenocytes are then fused by well-known techniques to any suitable myeloma cells, for example, cells from cell line SP20 available from the American Type Culture Collection (ATCC®) (Manassas, VA), to form hybridomas. Hybridomas are selected and cloned by limited dilution. In certain embodiments, lymph nodes of the immunized mice are harvested and fused with NS0 myeloma cells.

[0201] The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells lack the enzyme hypoxanthineguanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT -deficient cells.

[0202] Specific embodiments employ myeloma cells that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these myeloma cell lines are murine myeloma lines, such as the NSO cell line or those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, CA, USA, and SP-2 or X63-Ag8.653 cells available from the American Type Culture Collection, Rockville, MD, USA. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor D (1984) J Immunol 133: 3001-5; Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987), each of which is herein incorporated by reference in its entirety).

[0203] Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against NAPRT (e.g, human NAPRT). The binding specificity of monoclonal antibodies produced by hybridoma cells is determined by methods known in the art, for example, immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).

[0204] After hybridoma cells are identified that produce antibodies of the desired specificity, affinity, and / or activity, the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding JW (Ed.), Monoclonal Antibodies: Principles and Practice, supra'). Suitable culture media for this purpose include, for example, D-MEM or RPMI 1640 medium. In addition, the hybridoma cells may be grown in vivo as ascites tumors in an animal.

[0205] The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0206] Antibodies described herein include, e.g., antibody fragments which recognize NAPRT (e.g., human NAPRT), and can be generated by any technique known to those of skill in the art. For example, Fab and F(ab’)2 fragments described herein can be produced by proteolytic cleavage of immunoglobulin molecules, using enzymes such as papain (to produce Fab fragments) or pepsin(to produce F(ab’)2 fragments). A Fab fragment corresponds to one of the two identical arms of an antibody molecule and contains the complete light chain paired with the VH and CHI domains of the heavy chain. A F(ab’)2 fragment contains the two antigen-binding arms of an antibody molecule linked by disulfide bonds in the hinge region.

[0207] Further, the antibodies described herein can also be generated using various phage display methods known in the art. In phage display methods, functional antibody domains are displayed on the surface of phage particles which carry the polynucleotide sequences encoding them. In particular, DNA sequences encoding VH and VL domains are amplified from animal cDNA libraries (e.g., human or murine cDNA libraries of affected tissues). The DNA encoding the VH and VL domains are recombined together with an scFv linker by PCR and cloned into a phagemid vector. The vector is electroporated in E. coli, and the E. coli is infected with helper phage. Phage used in these methods are typically filamentous phage, including fd and Ml 3, and the VH and VL domains are usually recombinantly fused to either the phage gene III or gene VIII. Phage expressing an antigen-binding region that binds to a particular antigen can be selected or identified with antigen, e.g., using labeled antigen or antigen bound or captured to a solid surface or bead. Examples of phage display methods that can be used to make the antibodies described herein include those disclosed in Brinkman U etal., (1995) J Immunol Methods 182: 41-50; Ames RS et al., (1995) J Immunol Methods 184: 177-186; Kettleborough CA et al., (1994) Eur J Immunol 24: 952-958; Persic L et al., (1997) Gene 187: 9-18; Burton DR & Barbas CF (1994) Advan Immunol 57: 191-280; PCT Application No. PCT / GB91 / 001134; International Publication Nos. WO 90 / 02809, WO 91 / 10737, WO 92 / 01047, WO 92 / 18619, WO 93 / 1 1236, WO 95 / 15982, WO 95 / 20401, and WO 97 / 13844; and U.S. Patent Nos. 5,698,426, 5,223,409, 5,403,484, 5,580,717, 5,427,908, 5,750,753, 5,821,047, 5,571,698, 5,427,908, 5,516,637, 5,780,225, 5,658,727, 5,733,743, and 5,969,108, all of which are herein incorporated by reference in their entireties.

[0208] As described in the above references, after phage selection, the antibody coding regions from the phage can be isolated and used to generate whole antibodies, including human antibodies, or any other desired antigen-binding fragment, and expressed in any desired host, including mammalian cells, insect cells, plant cells, yeast, and bacteria, e.g., as described below. Techniques to recombinantly produce antibody fragments such as Fab, Fab’ and F(ab’)2 fragments can also be employed using methods known in the art such as those disclosed in PCT publication No. WO92 / 22324; Mullinax RL et al., (1992) BioTechniques 12(6): 864-9; Sawai H et al., (1995) Am J Reprod Immunol 34: 26-34; and Better M et al., (1988) Science 240: 1041-1043, all of which are herein incorporated by reference in their entireties.

[0209] In certain embodiments, to generate whole antibodies, PCR primers including VH or VL nucleotide sequences, a restriction site, and a flanking sequence to protect the restriction site, can be used to amplify the VH or VL sequences from a template, e.g., scFv clones. Utilizing cloning techniques known to those of skill in the art, the PCR amplified VH domains can be cloned into vectors expressing a VH constant region, and the PCR amplified VL domains can be cloned into vectors expressing a VL constant region, e.g., human kappa or lambda constant regions. The VH and VL domains can also be cloned into one vector expressing the necessary constant regions. The heavy chain conversion vectors and light chain conversion vectors are then co-transfected into cell lines to generate stable or transient cell lines that express full-length antibodies, e.g., IgG, using techniques known to those of skill in the art.

[0210] A chimeric antibody is a molecule in which different portions of the antibody are derived from different immunoglobulin molecules. For example, a chimeric antibody can contain a variable region of a mouse or rat monoclonal antibody fused to a constant region of a human antibody. Methods for producing chimeric antibodies are known in the art. See, e.g., Morrison SL (1985) Science 229: 1202-7; Oi VT & Morrison SL (1986) BioTechniques 4: 214-221; Gillies SD et al., (1989) J Immunol Methods 125: 191-202; and U.S. Patent Nos. 5,807,715, 4,816,567, 4,816,397, and 6,331,415, all of which are herein incorporated by reference in their entireties.

[0211] A humanized antibody is capable of binding to a predetermined antigen and which comprises a framework region having substantially the amino acid sequence of a human immunoglobulin and CDRs having substantially the amino acid sequence of a non-human immunoglobulin (e.g., a murine immunoglobulin). In certain embodiments, a humanized antibody also comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. The antibody also can include the CHI, hinge, CH2, CH3, and CH4 regions of the heavy chain. A humanized antibody can be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA, and IgE, and any isotype, including IgGi, IgG2, IgG.3, and IgG4. Humanized antibodies can be produced using a variety of techniques known in the art, including but not limited to, CDR-grafting (European Patent No. EP 239400; International Publication No. WO 91 / 09967; and U.S. Patent Nos. 5,225,539, 5,530,101, and 5,585,089),veneering or resurfacing (European Patent Nos. EP 592106 and EP 519596; Padlan EA (1991) Mol Immunol 28(4 / 5): 489-498; Studnicka GM et al., (1994) Prot Engineering 7(6): 805-814; and Roguska MA et al., (1994) PNAS 91 : 969-973), chain shuffling (U.S. Patent No. 5,565,332), and techniques disclosed in, e.g., U.S. Pat. No. 6,407,213, U.S. Pat. No. 5,766,886, International Publication No. WO 93 / 17105; Tan P et al., (2002) J Immunol 169: 1119-25; Caldas C et al., (2000) Protein Eng. 13(5): 353-60; Morea V et al., (2000) Methods 20(3): 267-79; Baca M et al., (1997) I Biol Chem 272(16): 10678-84; Roguska MA et al., (1996) Protein Eng 9(10): 895 904; Couto JR et al., (1995) Cancer Res. 55 (23 Supp): 5973s-5977s; Couto JR el al., (1995) Cancer Res 55(8): 1717-22; Sandhu JS (1994) Gene 150(2): 409-10; and Pedersen JT etal., (1994) J Mol Biol 235(3): 959-73, all of which are herein incorporated by reference in their entireties. See also, U.S. Application Publication No. US 2005 / 0042664 Al (Feb. 24, 2005), which is herein incorporated by reference in its entirety.

[0212] Methods for making multispecific antibodies (e.g., bispecific antibodies) have been described, see, e.g., U.S. Patent Nos. 7,951,917; 7,183,076; 8,227,577; 5,837,242; 5,989,830; 5,869,620; 6,132,992; and 8,586,713, all of which are herein incorporated by reference in their entireties.

[0213] Bispecific, bivalent antibodies, and methods of making them, are described, for instance in U.S. Pat. Nos. 5,731,168; 5,807,706; 5,821,333; and U.S. Appl. Publ. Nos. 2003 / 020734 and 2002 / 0155537, each of which is herein incorporated by reference in its entirety. Bispecific tetravalent antibodies, and methods of making them are described, for instance, in Int. Appl. Publ. Nos. WO 02 / 096948 and WO 00 / 44788, the disclosures of both of which are herein incorporated by reference in its entirety. See generally, Int. Appl. Publ. Nos. WO 93 / 17715, WO 92 / 08802, WO 91 / 00360, and WO 92 / 05793; Tutt et al., J. Immunol. 147:60-69 (1991); U.S. Pat. Nos. 4,474,893; 4,714,681; 4,925,648; 5,573,920; and 5,601,819; and Kostelny et al., J. Immunol. 148: 1547-1553 (1992); each of which is herein incorporated by reference in its entirety.

[0214] A bispecific antibody as described herein can be generated according to the DuoBody technology platform (Genmab A / S) as described, e.g., in International Publication Nos. WO 2011 / 131746, WO 2011 / 147986, WO 2008 / 119353, and WO 2013 / 060867, and in Labrijn AF et al., (2013) PNAS 110(13): 5145-5150. The DuoBody technology can be used to combine one half of a first monospecific antibody, or first antigen-binding region, containing two heavy and two light chains with one half of a second monospecific antibody, or second antigen-binding region,containing two heavy and two light chains. The resultant heterodimer contains one heavy chain and one light chain from the first antibody, or first antigen-binding region, paired with one heavy chain and one light chain from the second antibody, or second antigen-binding region. When both of the monospecific antibodies, or antigen-binding regions, recognize different epitopes on different antigens, the resultant heterodimer is a bispecific antibody.

[0215] The DuoBody technology requires that each of the monospecific antibodies, or antigenbinding regions, includes a heavy chain constant region with a single point mutation in the CH3 domain. The point mutations allow for a stronger interaction between the CH3 domains in the resultant bispecific antibody than between the CH3 domains in either of the monospecific antibodies, or antigen-binding regions. The single point mutation in each monospecific antibody, or antigen-binding region, is at residue 366, 368, 370, 399, 405, 407, or 409, numbered according to the EU numbering system, in the CEB domain of the heavy chain constant region, as described, e.g., in International Publication No. WO 2011 / 131746. Moreover, the single point mutation is located at a different residue in one monospecific antibody, or antigen-binding region, as compared to the other monospecific antibody, or antigen-binding region. For example, one monospecific antibody, or antigen-binding region, can comprise the mutation F405L (i.e., a mutation from phenylalanine to leucine at residue 405), while the other monospecific antibody, or antigenbinding region, can comprise the mutation K409R (i.e., a mutation from lysine to arginine at residue 409), numbered according to the EU numbering system. The heavy chain constant regions of the monospecific antibodies, or antigen-binding regions, can be an IgGi, IgG2, IgGi, or IgG4 isotype (e.g., a human IgGi isotype), and a bispecific antibody produced by the DuoBody technology can retain Fc-mediated effector functions.

[0216] Another method for generating bispecific antibodies has been termed the “knobs-into- holes” strategy (see, e.g., Inti. Publ. W02006 / 028936). The mispairing of Ig heavy chains is reduced in this technology by mutating selected amino acids forming the interface of the CH3 domains in IgG. At positions within the CH3 domain at which the two heavy chains interact directly, an amino acid with a small side chain (hole) is introduced into the sequence of one heavy chain and an amino acid with a large side chain (knob) into the counterpart interacting residue location on the other heavy chain. In certain embodiments, compositions of the disclosure have immunoglobulin chains in which the CH3 domains have been modified by mutating selected amino acids that interact at the interface between two polypeptides so as to preferentially form abi specific antibody. The bispecific antibodies can be composed of immunoglobulin chains of the same subclass (e.g., IgGi or IgGa) or different subclasses (e.g., IgGi and IgGa, or IgGa and IgG4).

[0217] Bispecific antibodies can, in certain instances contain, IgG4 and IgGi, IgG4 and IgGa, IgG4and IgGa, or IgGi and IgGa chain heterodimers. Such heterodimeric heavy chain antibodies can routinely be engineered by, for example, modifying selected amino acids forming the interface of the CH3 domains in human IgG4 and the IgGi or IgGa, so as to favor heterodimeric heavy chain formation.

[0218] In certain embodiments, an antibody described herein, which binds to the same epitope of NAPRT (e.g., human NAPRT) as an anti-NAPRT (e.g, human NAPRT) antibody described herein, is a human antibody. In certain embodiments, an antibody described herein, which competitively blocks (e.g, in a dose-dependent manner) any one of the antibodies described herein, from binding to NAPRT (e.g., human NAPRT), is a human antibody. Human antibodies can be produced using any method known in the art. For example, transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which can express human immunoglobulin genes, can be used. In particular, the human heavy and light chain immunoglobulin gene complexes can be introduced randomly or by homologous recombination into mouse embryonic stem cells. Alternatively, the human variable region, constant region, and diversity region can be introduced into mouse embryonic stem cells in addition to the human heavy and light chain genes. The mouse heavy and light chain immunoglobulin genes can be rendered non-functional separately or simultaneously with the introduction of human immunoglobulin loci by homologous recombination. In particular, homozygous deletion of the JH region prevents endogenous antibody production. The modified embryonic stem cells are expanded and microinjected into blastocysts to produce chimeric mice. The chimeric mice are then bred to produce homozygous offspring which express human antibodies. The transgenic mice are immunized in the normal fashion with a selected antigen, e.g., all or a portion of an antigen (e.g., NAPRT (e.g., human NAPRT)). Monoclonal antibodies directed against the antigen can be obtained from the immunized, transgenic mice using conventional hybridoma technology. The human immunoglobulin transgenes harbored by the transgenic mice rearrange during B cell differentiation, and subsequently undergo class switching and somatic mutation. Thus, using such a technique, it is possible to produce therapeutically useful IgG, IgA, IgM, and IgE antibodies. For an overview of this technology for producing human antibodies, see Lonberg N & Huszar D (1995)Int Rev Immunol 13: 65-93, herein incorporated by reference in its entirety. For a detailed discussion of this technology for producing human antibodies and human monoclonal antibodies and protocols for producing such antibodies, see, e.g., International Publication Nos. WO 98 / 24893, WO 96 / 34096, and WO 96 / 33735; and U.S. Patent Nos. 5,413,923, 5,625,126, 5,633,425, 5,569,825, 5,661,016, 5,545,806, 5,814,318, and 5,939,598, all of which are herein incorporated by reference in their entireties. Examples of mice capable of producing human antibodies include the XenoMouse™ (Abgenix, Inc.; U.S. Patent Nos. 6,075,181 and 6,150,184), the HuAb-Mouse™ (Medarex, Inc. / Gen Pharm; U.S. Patent Nos. 5,545,806 and 5,569, 825), the TransChromo Mouse™ (Kirin) and the KM Mouse™ (Medarex / Kirin), all of which are herein incorporated by reference in their entireties.

[0219] Human antibodies that specifically bind to NAPRT (e.g. , human NAPRT) can be made by a variety of methods known in the art, including the phage display methods described above using antibody libraries derived from human immunoglobulin sequences. See also, U.S. Patent Nos. 4,444,887, 4,716, 111, and 5,885,793; and International Publication Nos. WO 98 / 46645, WO 98 / 50433, WO 98 / 24893, WO 98 / 16654, WO 96 / 34096, WO 96 / 33735, and WO 91 / 10741, all of which are herein incorporated by reference in their entireties.

[0220] In certain embodiments, human antibodies can be produced using mouse-human hybridomas. For example, human peripheral blood lymphocytes transformed with Epstein-Barr virus (EBV) can be fused with mouse myeloma cells to produce mouse-human hybridomas secreting human monoclonal antibodies, and these mouse-human hybridomas can be screened to determine ones which secrete human monoclonal antibodies that specifically bind to a target antigen (e.g., NAPRT (e.g., human NAPRT)). Such methods are known and are described in the art, see, e.g., Shinmoto H et al., (2004) Cytotechnology 46: 19-23; Naganawa Y et al., (2005) Human Antibodies 14: 27-31, each of which is herein incorporated by reference in its entirety.Kits

[0221] Also provided are kits comprising one or more antibodies described herein, or pharmaceutical compositions or conjugates thereof. In a specific embodiment, provided herein is a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions described herein, such as one or more antibodies provided herein. In certain embodiments, the kits contain a pharmaceutical composition described herein and any prophylactic or therapeutic agent, such as those described herein. In certainembodiments, the kits may contain a T cell mitogen, such as, e.g., phytohaemagglutinin (PHA) and / or phorbol myristate acetate (PMA), or a TCR complex stimulating antibody, such as an anti- CD3 antibody and anti-CD28 antibody. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.

[0222] Also provided are kits that can be used in the above methods. In certain embodiments, a kit comprises an antibody described herein, preferably purified antibody, in one or more containers. In a specific embodiment, kits described herein contain a substantially isolated NAPRT (e.g., human NAPRT) antigen as a control. In another specific embodiment, the kits described herein further comprise a control antibody which does not react with NAPRT (e.g., human NAPRT) antigen. In another specific embodiment, kits described herein contain one or more elements for detecting the binding of an antibody to an NAPRT (e.g., human NAPRT) antigen (e.g., the antibody can be conjugated to a detectable substrate such as a fluorescent compound, an enzymatic substrate, a radioactive compound or a luminescent compound, or a second antibody which recognizes the first antibody can be conjugated to a detectable substrate). In specific embodiments, a kit provided herein can include a recombinantly produced or chemically synthesized NAPRT (e.g., human NAPRT) antigen. The NAPRT (e.g., human NAPRT) antigen provided in the kit can also be attached to a solid support. In a more specific embodiment, the detecting means of the above-described kit includes a solid support to which an NAPRT (e.g., human NAPRT) antigen is attached. Such a kit can also include a non-attached reporter-labeled anti-human antibody or anti-mouse / rat antibody. In this embodiment, binding of the antibody to the NAPRT (e.g., human NAPRT) antigen can be detected by binding of the said reporter-labeled antibody. In certain embodiments, the present disclosure relates to the use of a kit of the present disclosure for in vitro assaying and / or detecting NAPRT (e.g., human NAPRT) antigen in a biological sample.EXAMPLES

[0223] The following examples are offered by way of illustration and not by way of limitation.Example 1 ; Generation and characterization of NAPRT antibodies

[0224] Monoclonal, NAPRT-specific, antibodies were generated using human NAPRTl(e3) (SEQ ID NO: 31) as the immunogen in mouse.

[0225] The affinity of the NAPRT-specific antibodies for NAPRTl(e3)-1861 (SEQ ID NO: 40) was measured using enzyme-linked immunosorbent assay (ELISA). Several antigens were used for this assay, as summarized in the Table 7 below.Table 7. Antigens used for ELISA assay.

[0226] The antigens were coated at an amount of lOOpg / well. at 4°C overnight. Blocking was subsequently performed with 1%BSA / PBS, 200pL / well, at 4°C overnight.

[0227] For performing the assay, the primary antibodies used were obtained by centrifugation from hybridoma subclones supernatant. l OOpL of the supernatant of a single subclone was added to single well of the assay, and the assay plate was then incubated at 37°C for 1 hour. A positive control comprising NAPRT(e3)-1861 Anti-serum (dilution 1 : 1000) and a negative control comprising PBS only were used, both at a volume of lOOpL / well.

[0228] After incubation, a secondary antibody, HRP labeled anti-Mouse IgG (SIGMA, Cat#: A0168, Lot NO: # 068M4764V) was added to each well (dilution: 1 :9,000, 50pL / well) and incubated at 37°C for 1 hour. Finally, the substrate solution (TMB (3, 3', 5, 5'-Tetramethylbenzidine)) was added at an amount of, l OOpL / well, and incubated at 37°C for lOmin. Optical density (OD) was subsequently read at 450nm. Table 8 below shows the results of the ELISA experiment.Table 8. Optical density (OD) values of NAPRT-specific antibodies obtained by ELISA.

[0229] Based on this experiments, clones 2G11A3 and 4A5D7 showed the highest binding affinity for Antigens 1, 4 and 5, and thus the highest affinity to human NAPRT.

[0230] These two clones showed effective binding to NAPRT in Western blot experiments as showed in FIG. 1. In these Western blot experiments, NAPRT was indeed detected in samples from the NAPRT -positive human colorectal carcinoma cell line HCT116, and not detected in theNAPRT negative U2OS cell line. The 4A5D7 antibody appeared to show the highest binding affinity to human NAPRT.Example 2; Detection of NAPRT in different cancer cells

[0231] The 4A5D7 antibody was used to characterize samples obtain subjects suffering from different cancer, with regard to the presence of detectable NAPRT. Table 9 below presents the results of this study.Table 9. Summary of NAPRT detection in different cancersExample 3; NAMPTi are Effective in NAPRT Negative Patient-Derived Tumors

[0232] Three NAMPTis, ATN-249, ATN-940, and ATN-1083, were tested for their respective efficacy in two patient-derived colorectal adenocarcinoma ex vivo models, one of which is NAPRT -positive and the other of which is NAPRT -negative. NAPRT status of these models wasidentified by immunohistochemistry (FIG. 2D). As shown in FIGs. 2A-2C, these three NAMPTis were effective in reducing the viability ratio in this ex vivo model.

[0233] The effectiveness of ATN-249 was then tested across a variety of other NAPRT- negative and NAPRT-positive patient-derived tumors ex vivo. As can be seen in FIG. 3A, ATN-249 had no or limited effect on NAPRT-positive tumor models, but noticeable effects on NAPRT - negative models as shown in FIG. 3B.

[0234] These NAPRT-negative and NAPRT-positive models were identified as such by immunohistochemistry (IHC) or by Western blot (WB). The responsiveness of these models to ATN-249 are shown in the tables below, where a “response” to NAMPTi is defined as an IC50 inferior or equal to 125nM, with an observed reduction in viability (“killing”) of at least 55%.Table 10: Responsiveness of models identified by IHC.Table 11: Responsiveness of models identified by WB.

[0235] The values of Table 10 gave a positive predictive value (PPV) of 100%, a negative predictive value (NPV) of 48%, and an overall predictive value (OPV) of 65%. However, the values of Table 11 gave a PPV of 92%, an NPV of 87%, and thus an OPV of 88%. Upon investigation using WB, it was determined that a subset of NAPRT-negative responders identified by IHC alone had low level expression of NAPRT.

[0236] The effectiveness of ATN-940 in inhibiting growth of colorectal cancer in vivo was also ascertained, for both the NAPRT-negative model and the NAPRT-positive model. In this experiment, ATN-940 was administered at 1 mg of base / kg of body weight (mpk), once daily(QD), on days 1-3, 8-10, 15-17, and 22-24, and change in tumor volume in the subject comprising the xenograft was recorded over the span of 30 days. As shown in Fig. 4, ATN-940, ATN-940 inhibited growth of the NAPRT -negative tumor when compared to the administration of a vehicle. ATN-940 however did not prevent the growth of the NAPRT-positive tumor.* * *

[0237] The invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0238] All references (e. , publications or patents or patent applications) cited herein are incorporated herein by reference in their entireties and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes.

[0239] Other embodiments are within the following claims.INFORMAL SEQUENCE LISTING

[0240] VH amino acid sequence of antibody 4A5D7:EVQLQQSGPELVKPGASVKISCKTSGYTFTEYTMHWVKQSHGKSLEWIGGIYPNNYDTNYSQKFKGKATLTVDKSSSTAYMELRSLTSEDSAVYYCARTVYYAMDYWGQGTSVTVSS (SEQ ID NO: 1)

[0241] VL amino acid sequence of antibody 4A5D7:DIVITQDELSNPVTSGESVSISCRSSKSLLYKDGKTYLNWLLQRPGQSPQLLIYLMSTRASGVSDRFSGSGSGTDFTLEISRVKAEDVGVYYCQQLVDYPLTFGAGTKLELKR

[0242] CDRH1 amino acid sequence of sequence antibody 4A5D7 (Kabat):EYTMH (SEQ ID NO: 3)

[0243] CDRH2 amino acid sequence of sequence antibody 4A5D7 (Kabat):GIYPNNYDTNYSQKFKG (SEQ ID NO: 4)

[0244] CDRH3 amino acid sequence of sequence antibody 4A5D7 (Kabat):TVYYAMDY (SEQ ID NO: 5)

[0245] CDRH1 amino acid sequence of sequence antibody 4A5D7 (Chothia):GYTFTEY (SEQ ID NO: 9)

[0246] CDRH2 amino acid sequence of sequence antibody 4A5D7 (Chothia):YPNNYD (SEQ ID NO: 10)

[0247] CDRH3 amino acid sequence of sequence antibody 4A5D7 (Chothia):TVYYAMDY (SEQ ID NO: 5)

[0248] CDRH1 amino acid sequence of sequence antibody 4A5D7 (IMGT):GYTFTEYT (SEQ ID NO: 11)

[0249] CDRH2 amino acid sequence of sequence antibody 4A5D7 (IMGT):IYPNNYDT (SEQ ID NO: 12)

[0250] CDRH3 amino acid sequence of sequence antibody 4A5D7 (IMGT):ARTVYYAMDY (SEQ ID NO: 13)

[0251] CDRH1 amino acid sequence of sequence antibody 4A5D7 (AbM):GYTFTEYTMH (SEQ ID NO: 15)

[0252] CDRH2 amino acid sequence of sequence antibody 4A5D7 (AbM):GIYPNNYDTN (SEQ ID NO: 16)

[0253] CDRH3 amino acid sequence of sequence antibody 4A5D7 (AbM):TVYYAMDY (SEQ ID NO: 5)

[0254] CDRH1 amino acid sequence of sequence antibody 4A5D7 (MacCallum):TEYTMH (SEQ ID NO: 17)

[0255] CDRH2 amino acid sequence of sequence antibody 4A5D7 (MacCallum): WIGGIYPNNYDTN (SEQ ID NO: 18)

[0256] CDRH3 amino acid sequence of sequence antibody 4A5D7 (MacCallum):ARTVYYAMD (SEQ ID NO: 19)

[0257]

[0258] CDRL1 amino acid sequence of sequence antibody 4A5D7 (Kabat):RSSKSLLYKDGKTYLN (SEQ ID NO: 6)

[0259] CDRL2 amino acid sequence of sequence antibody 4A5D7 (Kabat):LMSTRAS (SEQ ID NO: 7)

[0260] CDRL3 amino acid sequence of sequence antibody 4A5D7 (Kabat):QQLVDYPLT (SEQ ID NO: 8)

[0261] CDRL1 amino acid sequence of sequence antibody 4A5D7 (Chothia):RSSKSLLYKDGKTYLN (SEQ ID NO: 6)

[0262] CDRL2 amino acid sequence of sequence antibody 4A5D7 (Chothia):LMSTRAS (SEQ ID NO: 7)

[0263] CDRL3 amino acid sequence of sequence antibody 4A5D7 (Chothia):QQLVDYPLT (SEQ ID NO: 8)

[0264] CDRL1 amino acid sequence of sequence antibody 4A5D7 (IMGT):KSLLYKDGKTY (SEQ ID NO: 14)

[0265] CDRL2 amino acid sequence of sequence antibody 4A5D7 (IMGT):LM or LMS

[0266] CDRL3 amino acid sequence of sequence antibody 4A5D7 (IMGT):QQLVDYPLT (SEQ ID NO: 8)

[0267] CDRL1 amino acid sequence of sequence antibody 4A5D7 (AbM):RSSKSLLYKDGKTYLN (SEQ ID NO: 6)

[0268] CDRL2 amino acid sequence of sequence antibody 4A5D7 (AbM):LMSTRAS (SEQ ID NO: 7)

[0269] CDRL3 amino acid sequence of sequence antibody 4A5D7 (AbM):QQLVDYPLT (SEQ ID NO: 8)

[0270] CDRL1 amino acid sequence of sequence antibody 4A5D7 (MacCallum): LYKDGKTYLNWL (SEQ ID NO: 20)

[0271] CDRL2 amino acid sequence of sequence antibody 4A5D7 (MacCallum): LLIYLMSTRA (SEQ ID NO: 21)

[0272] CDRL3 amino acid sequence of sequence antibody 4A5D7 (MacCallum): QQLVDYPLT (SEQ ID NO: 8)

[0273] Nucleic acid sequence of VH of antibody 4A5D7:GAGGTCCAGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAA GATATCCTGCAAGACTTCTGGATACACATTCACTGAATACACCATGCACTGGGTGAA GCAGAGCCATGGAAAGAGCCTTGAGTGGATTGGAGGTATTTATCCTAACAATTATG ATACTAACTACAGCCAGAAATTCAAGGGCAAGGCCACATTGACTGTAGACAAGTCC TCCAGCACAGCCTACATGGAACTCCGCAGCCTGACATCTGAGGATTCTGCAGTCTAT TACTGTGCAAGAACGGTTTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTC ACCGTCTCCTCA (SEQ ID NO: 22)

[0274] Nucleic acid sequence of VL of antibody 4A5D7:GATATTGTGATAACCCAGGATGAACTCTCCAATCCTGTCACTTCTGGAGAATCAGTT TCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTATATAAGGATGGGAAGACATACTTG AATTGGTTGCTGCAGAGACCAGGACAATCTCCTCAGCTCCTGATCTATTTGATGTCC ACCCGTGCATCAGGAGTCTCAGACCGGTTTAGTGGCAGTGGGTCAGGAACAGATTTC ACCCTGGAAATCAGTAGAGTGAAGGCTGAGGATGTGGGTGTGTATTACTGTCAACA ACTTGTAGATTATCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGG (SEQ ID NO: 23)

[0275] WT IgGl heavy chain constant region:ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GL YSL S S VVTVP S S SLGTQTYICNVNHKP SNTKVDKKVEPKSCDKTHTCPPCP APELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR DELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 24)

[0276] S228P IgG4 heavy chain constant region:ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GL YSL S S VVTVP S S SLGTKTYTCNVDHKP SNTKVDKRVESKYGPPCPPCP APEFLGGP S V FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNST YRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEM TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRW QEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 25)

[0277] Kappa light chain constant region:RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 26)

[0278] Lambda light chain constant region:GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPS KQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 27)

[0279] Antigen 3 :LAEEPVPQAGAEC (SEQ ID NO: 28)

[0280] Antigen 4:AGQELRVWPPGAC (SEQ ID NO: 29)

[0281] Antigen 5:LRVWPPGAQEPC (SEQ ID NO: 30)

[0282] Amino acid sequence of human NAPRT1 (e3):HLGLGVQEPHPGERAAFVAYALAFPRAFQGLLDTYSVWRSGLPNFLAVAL LGELGYR AVGVRLDSGDLLQQAQEIRKVFRAAAAQFQVPWLESVLIVVSNNIDEEALARLAQEGSE VNVIGIGTSVVTCPQQPSLGGVYKLVAVGGQPRMKLTEDPEKQTLPGSKAAFRLLGSDG SPLMDMLQLAEEPVPQAGQELRVWPPGAQEPCTVRPAQVEPLLRLCLQQGQLCEPLPSL AESRALAQLSLSRLSPEHRRLRSPALEHHHHHH (SEQ ID NO: 31)

[0283] Nucleic acid sequence of human NAPRT1 (e3):CACCTGGGGCTGGGGGTGCAGGAGCCGCATCCAGGCGAGCGGGCAGCCTTTGTGGC CTATGCCTTGGCTTTTCCCCGGGCCTTCCAGGGCCTCCTGGACACCTACAGCGTGTG GAGGAGTGGTCTCCCCAACTTCCTAGCAGTCGCCCTGGCCCTGGGAGAGCTGGGCTA CCGGGCAGTGGGCGTGAGGCTGGACAGTGGTGACCTGCTACAGCAGGCTCAGGAGA TCCGCAAGGTCTTCCGAGCTGCTGCAGCCCAGTTCCAGGTGCCCTGGCTGGAGTCAG TCCTCATCGTAGTCAGCAACAACATTGACGAGGAGGCGCTGGCCCGACTGGCCCAGGAGGGCAGTGAGGTGAATGTCATTGGCATTGGCACCAGTGTGGTCACCTGCCCCCA ACAGCCTTCCCTGGGTGGCGTCTATAAGCTGGTGGCCGTGGGGGGCCAGCCACGAA TGAAGCTGACCGAGGACCCCGAGAAGCAGACGTTGCCTGGGAGCAAGGCTGCTTTC CGGCTCCTGGGCTCTGACGGGTCTCCACTCATGGACATGCTGCAGTTAGCAGAAGAG CCAGTGCCACAGGCTGGGCAGGAGCTGAGGGTGTGGCCTCCAGGGGCCCAGGAGCC CTGCACCGTGAGGCCAGCCCAGGTGGAGCCACTACTGCGGCTCTGCCTCCAGCAGG GACAGCTGTGTGAGCCGCTCCCATCCCTGGCAGAGTCTAGAGCCTTGGCCCAGCTGT CCCTGAGCCGACTCAGCCCTGAGCACAGGCGGCTGCGGAGCCCTGCACTCGAGCACCACCACCACCACCACTGA(SEQ ID NO: 32)

[0284] Amino acid sequence of NAPRT encoded by transcript “NAPRT-212” (UniProtKB accession number: H0YF31):ARDAAEFELFFRRCPFGGAFALAAGLRDCVRFLRAFRLRDADVQFLASVLPPDTDPAFF EHLRALDCSEVTVRALPEGSLAFPGVPLLQVSGPLLVVQLLETPLLCLVSYASLVATNAA RLRLIAGPEKRLLEMGLRRAQGPDGGLTASTYSYLGDVGASSW (SEQ ID NO: 33)

[0285] Amino acid sequence of NAPRT encoded by transcript “NAPRT-214” (UniProtKB accession number: H0YDA6):ALAAGLRDCVRFLRAFRLRDADVQFLASVLPPDTDPAFFEHLRALDCSEVPLLQVSGPL LWQLLETPLLCLVSYASLVATNAARLRLIAGPEKRLLEMGLRRAQGPDGGLTASTYSY LGDVGASSW (SEQ ID NO: 34)

[0286] Amino acid sequence of NAPRT encoded by transcript “NAPRT-205” (UniProtKB accession number: A0A087WUT5):FQVPWLESVLIVVSNNIDEEALARLAQEGSEVNVIGIGTSVVTCPQQPSLGGVYKLVAVG GQPRMKLTEDPEKQTLPGSKAAFRLLGSDGSPLMDMLQLAEEPVPQAGQELRVWPPGA QEPCTVRPAQLCEPLPSLAESRALAQLSLSRLSPEHRRLRSPAQYQVGGRPTLSFCPVRPR PHPAHRSCPLLPAGGAVREAAGPGEQSVCGAVPLRLGAGLTGNNTNHSLFPT (SEQ ID NO: 35)

[0287] Canonical Sequence of human NAPRT (UniProtKB accession number: Q6XQN6-1; NCBI Reference Sequence: NP_660202.3; 538 AA; 57,578 Daltons; NMJ45201.6):MAAEQDPEARAAARPLLTDLYQATMALGYWRAGRARDAAEFELFFRRCPFGGAFALA AGLRDCVRFLRAFRLRDADVQFLASVLPPDTDPAFFEHLRALDCSEVTVRALPEGSLAFP GVPLLQVSGPLLVVQLLETPLLCLVSYASLVATNAARLRLIAGPEKRLLEMGLRRAQGPDGGLTASTYSYLGGFDSSSNVLAGQLRGVPVAGTLAHSFVTSFSGSEVPPDPMLAPAAG EGPGVDLAAKAQVWLEQVCAHLGLGVQEPHPGERAAFVAYALAFPRAFQGLLDTYSV WRSGLPNFLAVALALGELGYRAVGVRLDSGDLLQQAQEIRKVFRAAAAQFQVPWLESV LIVVSNNIDEE ALARL AQEGSE VNVIGIGT S VVTCPQQP SLGGVYKL VA VGGQPRMKLT EDPEKQTLPGSKAAFRLLGSDGSPLMDMLQLAEEPVPQAGQELRVWPPGAQEPCTVRP AQ VEPLLRLCLQQGQLCEPLP SLAESRAL AQL SL SRL SPEHRRLRSP AQ YQ VVLSERLQ A LVNSLCAGQSP (SEQ ID NO: 36)

[0288] Isoform 1 of human NAPRT (UniProtKB accession number: Q6XQN6-2; 567 AA; 60,291 Daltons):MAAEQDPEARAAARPLLTDLYQATMALGYWRAGRARDAAEFELFFRRCPFGGAFALA AGLRDCVRFLRAFRLRDADVQFLASVLPPDTDPAFFEHLRALDCSEVTVRALPEGSLAFP GVPLLQVSGPLLVVQLLETPLLCLVSYASLVATNAARLRLIAGPEKRLLEMGLRRAQGP DGGLTASTYSYLGGFDSSSNVLAGQLRGVPVAGTLAHSFVTSFSGSEVPPDPMLAPAAG EGPGVDLAAKAQVWLEQVCAHLGLGVQEPHPGERAAFVAYALAFPRAFQGLLDTYSV WRSGLPNFLAVALALGELGYRAVGVRLDSGDLLQQAQEIRKVFRAAAAQFQVPWLESV LIVVSNNIDEE ALARL AQEGSEVNVIGIGT S VVTCPQQP SLGGVYKL VAVGGQPRMKLT EDPEKQTLPGSKAAFRLLGSDGSPLMDMLQLAEEPVPQAGQELRVWPPGAQEPCTVRP AQ VEPLLRLCLQQGQLCEPLP SLAE SRAL AQL SL SRL SPEHRRLRSP AQYQVGGGGPPCH SALCAPALTLPTAPVLCSLQVVLSERLQALVNSLCAGQSP (SEQ ID NO: 37)

[0289] Isoform 2 of human NAPRT (UniProtKB accession number: Q6XQN6-3; NCBI Reference Sequence: NP_001273758.1; 525 AA; 56,099 Daltons):MAAEQDPEARAAARPLLTDLYQATMALGYWRAGRARDAAEFELFFRRCPFGGAFALA AGLRDCVRFLRAFRLRDADVQFLASVLPPDTDPAFFEHLRALDCSEVTVRALPEGSLAFP GVPLLQVSGPLLVVQLLETPLLCLVSYASLVATNAARLRLIAGPEKRLLEMGLRRAQGP DGGLTASTYSYLGGFDSSSNVLAGQLRGVPVAGTLAHSFVTSFSGSEVPPDPMLAPAAG EGPGVDLAAKAQVWLEQVCAHLGLGVQEPHPGERAAFVAYALAFPRAFQGLLDTYSV WRSGLPNFLAVALALGELGYRAVGVRLDSGDLLQQAQEIRKVFRAAAAQFQVPWLESV LIVVSNNIDEEALARLAQEGSEVNVIGIGTSVVTCPQQPSLGGVYKL VAVGGQPRMKLT EDPEKQTLPGSKAAFRLLGSDGSPLMDMLQLAEEPVPQAGQELRVWPPGAQEPCTVRP AQLCEPLPSLAESRALAQLSLSRLSPEHRRLRSPAQYQVVLSERLQALVNSLCAGQSP (SEQ ID NO: 38)

[0290] Computationally mapped isoforms of human NAPRT:MAAEQDPEARAAARPLLTDLYQATMALGYWRAGRARDAAEFELFFRRCPFGGAFALA AGLRDCVRFLRAFRLRDADVQFLASVLPPDTDPAFFEHLRALDCSEVTVRALPEGSLAFP GVPLLQVSGPLLVVQLLETPLLCLVSYASLVATNAARLRLIAGPEKRLLEMGLRRAQGP DGGLTASTYSYLGGFDSSSNVLAGQLRGVPVAGTLAHSFVTSFSGSEVPPDPMLAPAAG EGPGVDLAAKAQVWLEQVCAHLGLGVQEPHPGERAAFVAYALAFPRAFQGLLDTYSV WRSGLPNFLAVALALGELGYRAVGVRLDSGDLLQQAQEIRKVFRAAAAQFQVPWLESV LIVVSNNIDEEALARL AQEGSEVNVIGIGT S VVTCPQQP SLGGVYKL VAVGGQPRMKLT EDPEKQTLPGSKAAFRLLGSDGSPLMDMLQLAEEPVPQAGQELRVWPPGAQEPCTVRP AQVEPLLRLCLQQGQVAAPPPSS (SEQ ID NO: 39)

[0291] NAPRTl(e3)-1861:MASMTDQQAEARAFLSEEMIAEFKAAFDMWDADGGGDISTKELGTVMRMLGQ NPTKEELDAIIEEVDEDGSGTIDFEEFLVMMVRQMKEDAKGKSEEELANCFRIFDKNAD GFIDIEELGEILRATGEHVIEEDIEDLMKDSDKNNDGRIDFDEFLKMMEGELGEAHEAEE VHEEAHHEEAHHAEAHHEEAHAHAEEVHEPEEAPEEEEKPRINGSHLGLGVQEPHPGER AAFVAYALAFPRAFQGLLDTYSVWRSGLPNFLAVALALGELGYRAVGVRLDSGDLLQQ AQEIRKVFRAAAAQFQVPWLESVLIVVSNNIDEEALARLAQEGSEVNVIGIGTSVVTCPQ QPSLGGVYKLVAVGGQPRMKLTEDPEKQTLPGSKAAFRLLGSDGSPLMDMLQLAEEPV PQAGQELRVWPPGAQEPCTVRPAQVEPLLRLCLQQGQLCEPLPSLAESRALAQLSLSRLS PEHRRLRSPALEHHHHHH (SEQ ID NO: 40)

Claims

WHAT IS CLAIMED:

1. An antibody that specifically binds human nicotinic acid phosphoribosyltransferase (NAPRT), the antibody comprising: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NO: 1; and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NO: 2.

2. The antibody of claim 1, wherein the antibody comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13;15, 16, and 5; or 17, 18, and 19, respectively.

3. The antibody of claim 2, wherein the antibody comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21, and 8, respectively.

4. The antibody of any one of claims 1-3, wherein the antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 3, 4, 5, 6, 7, and 8; 9, 10, 5, 6, 7, and 8; 11, 12, 13, 14, LM, and 8; 11, 12, 13, 14, LMS, and 8; 15,16, 5, 6, 7, and 8; 17, 18, 19, 20, 21, and 8, respectively.

5. The antibody of any one of claims 1-4, wherein the VH comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 1 and wherein the VL comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 2.

6. The antibody of any one of claims 1-5, wherein the VH comprises the amino acid sequence of SEQ ID NO: 1 and wherein the VL comprises the amino acid sequence of SEQ ID NO: 2.

7. The antibody of any one of claims 1-6, wherein the antibody comprises a heavy chain constant region, selected from the group consisting of human IgGi, IgG2, IgGs, IgG4, IgAi, and IgA2.

8. The antibody of any one of claims 1 -7, wherein the antibody comprises a heavy chain constant region that is a variant of a wild-type heavy chain constant region, wherein the variant heavy chain constant region binds to an FcyR with lower affinity than the wild-type heavy chain constant region binds to the FcyR.

9. An antibody that cross-competes for binding to NAPRT with the antibody of any one of claims 1-8.

10. An antibody that specifically binds to the amino acid sequence of SEQ ID NO: 29 or 30.

11. A polypeptide comprising a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NO: 1.

12. The polypeptide of claim 10, wherein the VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13; 15, 16, and 5; or 17, 18, and 19, respectively.

13. A polypeptide comprising a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NO: 2.

14. The polypeptide of claim 12, wherein the VL comprises the CDRL1, CDRL2, and CDRL3 set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21, and 8, respectively.

15. A polypeptide comprising the amino acid sequence LM, LMS, or the amino acid sequences set forth in any one of SEQ ID NOs: 1-21.

16. A polynucleotide encoding: a VH, a VL, a heavy chain, and / or a light chain of an antibody of any one of claims 1-10; or a polypeptide of any one of claims 11-15.

17. A vector comprising the polynucleotide of claim 16.

18. A recombinant host cell comprising:(a) the polynucleotide of claim 16;(b) the vector of claim 17;(c) a first polynucleotide encoding a heavy chain variable region or a heavy chain of the antibody of any one of claims 1-10 and a second polynucleotide encoding a light chain variable region or a light chain of the antibody of any one of claims 1-10; and / or(d) a first vector comprising a first polynucleotide encoding a heavy chain variable region or a heavy chain of the antibody of any one of claims 1-10 and a second vector comprising a second polynucleotide encoding a light chain variable region or a light chain of the antibody of any one of claims 1-10.

19. A composition comprising the antibody of any one of claims 1-10, the polypeptide of any one of claims 11-15, the polynucleotide of claim 16, the vector of claim 17, or the host cell of claim 18, and a pharmaceutically acceptable carrier or excipient.

20. A method of producing an antibody, the method comprising culturing the host cell of claim 18 under suitable conditions such that the polynucleotide is expressed, and the antibody is produced.

21. A method of determining whether a sample comprising cells from a subject is nicotinic acid phosphoribosyltransferase (NAPRT) negative comprising administering an antibody that specifically binds to NAPRT to the sample and detecting specific binding of the antibody to NAPRT in less than 1% of the cells in the sample, thereby determining that the sample is NAPRT negative.

22. The method of claim 21, wherein the sample comprises cancer cells.

23. The method of claim 22, wherein the cancer cells are solid tumor cells.

24. The method of claim 23, wherein the cancer cells are thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectalcancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer cells.

25. A method of treating cancer comprising administering a nicotinamide phosphoribosyltransferase inhibitor (NAMPTi) to a subject in need thereof, wherein a sample from the cancer has lower NAPRT levels than a control sample.

26. The method of claim 25, further comprising administering niacin to the subject.

27. The method of claim 25 or 26, wherein the cancer is NAPRT negative.

28. The method of any one of claims 25-27, wherein the cancer is thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer, cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer.

29. A method of diagnosing a subject with a cancer with increased likelihood to be effectively treated with a NAMPTi comprising administering an antibody that specifically binds to NAPRT to a sample from the subject and detecting specific binding of the antibody to NAPRT, wherein if there is less binding to NAPRT in the sample from the subject than in a control sample, the cancer in the subject has an increased likelihood to be effectively treated with the NAMPTi.

30. The method of claim 29, wherein the cancer in the subject has an increased likelihood to be effectively treated with niacin and the NAMPTi.

31. The method of claim 29 or 30, wherein the cancer is thyroid cancer, duodenal, neuroendocrine carcinoma (NEC), uterine cancer, small cell lung cancer (SCLC), hepatocellular carcinoma, mesothelioma, breast cancer, sarcoma, ovarian cancer, renal cell carcinoma, rectal cancer, head and neck cancer, prostate cancer, pancreatic cancer, melanoma, colorectal cancer,cervix cancer, non-small cell lung cancer (NSCLC), cholangiocarcinoma, or endometrial cancer cells.

32. The method of any one of claims 29-31, wherein the sample from the subject is NAPRT negative.

33. A method of diagnosing a subject with sepsis comprising administering an antibody that specifically binds to NAPRT to a sample from the subject and detecting specific binding of the antibody to NAPRT, wherein if there is more binding to NAPRT in the sample from the subject than in a control sample, the subject has an increased likelihood to have sepsis.

34. The method of any one of claims 21-33, wherein the NAPRT comprises the amino acid sequence of SEQ ID NO: 29 or 30.

35. The method of any one of claims 21-34, wherein the antibody comprises: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence set forth in SEQ ID NO: 1; and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence set forth in SEQ ID NO: 2.

36. The method of claim 35, wherein the antibody comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences set forth in SEQ ID NOs: 3, 4, and 5; 9, 10, and 5; 11, 12, and 13; 15, 16, and 5; or 17, 18, and 19, respectively.

37. The method of claim 35 or 36, wherein the antibody comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs: 6, 7, and 8; 14, LM, and 8; 14, LMS, and 8; or 20, 21, and 8, respectively.

38. The method of any one of claims 35-37, wherein the antibody comprises the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences set forth in SEQ ID NOs:3, 4, 5, 6, 7, and 8; 9, 10, 5, 6, 7, and 8; 11 , 12, 13, 14, LM, and 8; 1 1, 12, 13, 14, LMS, and 8; 15, 16, 5, 6, 7, and 8; 17, 18, 19, 20, 21, and 8, respectively.

39. The method of any one of claims 35-38, wherein the VH comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 1 and wherein the VL rises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NOs: 2.

40. The antibody of any one of claims 35-39, wherein the VH comprises the amino acid sequence of SEQ ID NO: 1 and wherein the VL comprises the amino acid sequence of SEQ ID NO: 2.

41. The method of any one of claims 35-40, wherein the antibody comprises a heavy chain constant region, selected from the group consisting of human IgGi, IgG2, IgGs, IgG4, IgAi, and IgA2.

42. The antibody of any one of claims 35-41, wherein the antibody comprises a heavy chain constant region that is a variant of a wild-type heavy chain constant region, wherein the variant heavy chain constant region binds to an FcyR with lower affinity than the wild-type heavy chain constant region binds to the FcyR.

43. An antibody of any one of claims 1-10; the polypeptide of any one of claims 11-15; the polynucleotide of claim 16; the vector of claim 17; the host cell of claim 18; or the composition of claim 19, for use in medicine.