Anti-HIV vaccine antibody with reduced polyreactivity

Abstract

The present disclosure provides novel anti-HIV broadly neutralizing antibodies and antigen-binding fragments thereof. The disclosed anti-HIV antibodies exhibit improved biophysical properties, such as reduced polyreactivity and extended half-life, while retaining broad and potent neutralizing activity. The disclosed anti-HIV bNAb variants constitute a novel therapeutic strategy for treating and / or preventing HIV infection.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 62 / 871,393, filed on Jul. 8, 2019. The foregoing application is hereby incorporated by reference in its entirety.

[0002] (Statement Regarding Federally Sponsored Research or Development) This invention was made with government support under grants P01AI100148 and R01AI29784 awarded by the National Institutes of Health (NIH). The government has certain rights in this invention.

[0003] (Technical Field) The present invention relates to broad - spectrum and potent antibodies against human immunodeficiency virus (“HIV”).

Background Art

[0004] HIV causes acquired immunodeficiency syndrome (AIDS), a human condition characterized by clinical features such as wasting syndrome, central nervous system degeneration, and profound immunosuppression that leads to life - threatening opportunistic infections and malignancies. Since its discovery in 1981, human immunodeficiency virus type 1 (HIV - 1) has killed at least 25 million people worldwide. Even if the rate of HIV infection were to decrease by 2.5% per year, it is predicted that between 20 million and 60 million people will become infected over the next 20 years. There is a need for therapeutic agents and treatment methods for the treatment or inhibition of HIV infection.

[0005] Broadly neutralizing antibodies (bNAbs) against HIV are being developed as potential therapies for the treatment and / or prevention of HIV infection. Many bNAbs have been isolated and characterized from human donors. Some of these bNAbs have been modified to improve their breadth and / or potency of neutralization. One such engineered bNAb is NIH45-46, which has a heavy chain mutation G54W (NIH45-46 G54W). NIH45-46 G54W has a desirable neutralization profile, but it has biophysical properties such as polyreactivity / non-specific binding, short in vivo half-life, and a tendency to aggregate, which hinder its development as a human therapeutic. There are various known approaches to improving the biophysical properties of antibodies, but it can be difficult to improve these properties without sacrificing much of the antibody's neutralizing activity.

Summary of the Invention

[0006] In one aspect, the present disclosure provides a novel isolated anti-HIV broadly neutralizing antibody or an antigen-binding portion thereof, with improved biophysical properties such as reduced polyreactivity. The isolated anti-HIV antibody, or an antigen-binding portion thereof, comprises a heavy chain having one or more heavy chain substitutions at one or more residues selected from the group consisting of S5, S21, G54, G55, V57, T68, T70, V73, S75, F79, S82, D85, V89, Y97, and P112, numbered according to Kabat numbering, and having a heavy chain amino acid sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 1; or a light chain having one or more light chain substitutions at one or more residues selected from the group consisting of T5, S7, T10, S12, S14, T18, I20, S22, R24, Q27, S28, S30, R59, S61, S63, W65, D68, N70, S72, S74, and S78, and having a light chain amino acid sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 2.

[0007] In some embodiments, the isolated anti-HIV antibody or antigen-binding portion thereof has a heavy chain polypeptide sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 1, and the heavy chain amino acid sequence contains one or more heavy chain substitutions at one or more residues selected from the group consisting of S5, S21, G54, G55, V57, T68, T70, V73, S75, F79, S82, D85, V89, Y97, and P112; and a light chain amino acid sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 2, and the light chain amino acid sequence contains one or more light chain substitutions at one or more residues selected from the group consisting of T5, S7, T10, S12, S14, T18, I20, S22, R24, Q27, S28, S30, R59, S61, S63, W65, D68, N70, S72, S74, and S78.

[0008] In some embodiments, the one or more heavy chain substitutions are substitutions of S5D or S5E, or conservative substitutions of Asp or Glu at residue S5; substitutions of S21D or S21E, or conservative substitutions of Asp or Glu at residue S21; substitution of G54W or conservative substitution of Trp at residue G54; substitutions of G55D or G55E, or conservative substitutions of Asp or Glu at residue G55; substitutions of V57D or V57E, or conservative substitutions of Asp or Glu at residue V57; substitutions of T68D or T68E, or conservative substitutions of Asp or Glu at residue T68; substitutions of T70D or T70E, or conservative substitutions of Asp or Glu at residue T70; substitutions of V73D or V73E, or conservative substitutions of Asp or Glu at residue V73; substitutions of S75D or S75E, or conservative substitutions of Asp or Glu at residue S75; substitutions of F79D, F79E, F79Y or F79H, or conservative substitutions of Asp, Glu, Tyr or His at residue F79; substitutions of S82D or S82E, or conservative substitutions of Asp or Glu at residue S82; substitution of D85E, or conservative substitution of Asp at residue D85; substitutions of V89D or V89E, or conservative substitutions of Asp or Glu at residue V89; substitutions of Y97D or Y97E, or conservative substitutions of Asp or Glu at residue Y97; substitutions of P112D or P112E, or conservative substitutions of Asp or Glu at residue P112; or combinations thereof.

[0009] In some embodiments, the one or more light chain substitutions are a T5D or T5E substitution, or a conservative substitution of Asp or Glu at residue T5; an S7D or S7E substitution, or a conservative substitution of Asp or Glu at residue S7; a T10D or T10E substitution, or a conservative substitution of Asp or Glu at residue T10; an S12D or S12E substitution, or a conservative substitution of Asp or Glu at residue S12; an S14D or S14E substitution, or a conservative substitution of Asp or Glu at residue S14; a T18D or T18E substitution, or a conservative substitution of Asp or Glu at residue T18; an I20D or I20E substitution, or a conservative substitution of Asp or Glu at residue I20; an S22D or S22E substitution, or a conservative substitution of Asp or Glu at residue S22; an R24D or R24E substitution, or a conservative substitution of Asp or Glu at residue R24; a Q27D or Q27E substitution, or a conservative substitution of Asp or Glu at residue Q27; an S28Y or S28H substitution, or a conservative substitution of Tyr or His at residue S28; an S30D or S30E substitution, or a conservative substitution of Asp or Glu at residue S30; an R59D or R59E substitution, or a conservative substitution of Asp or Glu at residue R59; an S61D or S61E substitution, or a conservative substitution of Asp or Glu at residue S61; an S63D or S63E substitution, or a conservative substitution of Asp or Glu at residue S63; a W65D, W65E, W65Y or W65H substitution, or a conservative substitution of Asp, Glu, Tyr or His at residue W65; a D68E substitution, or a conservative substitution of Asp at residue D68; an N70D or N70E substitution, or a conservative substitution of Asp or Glu at residue N70; an S72D or S72E substitution, or a conservative substitution of Asp or Glu at residue S72; an S74D or S74E substitution, or a conservative substitution of Asp or Glu at residue S74; an S78D or S78E substitution, or a conservative substitution of Asp or Glu at residue S78; or a combination thereof.

[0010] In some embodiments, the one or more heavy chain substitutions further include the substitutions M428L and N434S.

[0011] In some embodiments, the light chain amino acid sequence comprises one or more light chain substitutions of (i) Q27E, S28H, and S30D; (ii) Q27E, S28H, S30D, and S74T; (iii) Q27E, S28H, S30D, S74T, M428L, and N434S; (iv) Q27E, S28Y, and S30D; (v) Q27D and S28H; (vi) S28Y; (vii) T5D, T10D, S12D, S14D, I20D, and S22D; or (viii) S61D, S63D, W65D, N70D, S72D, and S74D.

[0012] In some embodiments, the heavy chain amino acid sequence comprises one or more heavy chain substitutions including G54W and T68D; and the light chain amino acid sequence comprises one or more light chain substitutions of (a) Q27E, S28H, S30D, and S74T; (b) Q27E, S28H, S30D, S74T, M428L, and N434S; (c) Q27E, S28Y, and S30D; (d) Q27D and S28H; or (e) S28Y.

[0013] In some embodiments, the heavy chain amino acid sequence comprises one or more heavy chain substitutions including G54W; and the light chain amino acid sequence comprises one or more light chain substitutions of (a) Q27E, S28H, and S30D; (b) Q27E, S28Y, and S30D; or (c) Q27D and S28H.

[0014] In some embodiments, the heavy chain amino acid sequence comprises the amino acid sequences of SEQ ID NOs: 3 to 35. In some embodiments, the light chain amino acid sequence comprises the amino acid sequences of SEQ ID NO: 2 and SEQ ID NOs: 36 to 80.

[0015] In some embodiments, the isolated anti-HIV antibody comprises CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 comprising the respective sequences set forth in the set of CDR sequences of SEQ ID NO: 81, 84, 87, 90, 93, and 96; SEQ ID NO: 81, 84, 87, 99, 93, and 96; SEQ ID NO: 81, 84, 87, 100, 93, and 96; SEQ ID NO: 81, 84, 87, 101, 93, and 96; or SEQ ID NO: 81, 84, 87, 102, 93, and 96; (as indicated by Kabat numbering); SEQ ID NO: 82, 85, 88, 91, 94, and 97 (as indicated by IMGT numbering); or SEQ ID NO: 83, 86, 89, 92, 95, and 98 (as indicated by Chothia numbering).

[0016] In some embodiments, the heavy chain amino acid sequence and the light chain amino acid sequence comprise the respective amino acid sequences set forth in a sequence set selected from the group consisting of SEQ ID NO: 3 and 36; SEQ ID NO: 3 and 38; SEQ ID NO: 3 and 39; SEQ ID NO: 4 and 2; SEQ ID NO: 4 and 38; SEQ ID NO: 4 and 39; SEQ ID NO: 4 and 41; and SEQ ID NO: 34 and 37.

[0017] In some embodiments, the isolated anti-HIV antibody is a bispecific antibody comprising a first antigen-binding arm that binds to a first antigen and a second antigen-binding arm that binds to a second antigen, wherein the first antigen and the second antigen are different, and the first antigen-binding arm comprises the heavy chain amino acid sequence and the light chain amino acid sequence described above. In some embodiments, the second antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-l), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), CD160 (NK1), immune checkpoint proteins (e.g., PD-1, PD-L1, CTLA-4), and the second epitope of gp120.

[0018] The present disclosure also provides: (i) an isolated nucleic acid comprising a sequence encoding a CDR, a heavy chain variable region, or a light chain variable region of the aforementioned anti-HIV antibody, or an antigen-binding portion thereof; (ii) a vector comprising the aforementioned nucleic acid; and (iii) a cultured cell comprising the aforementioned vector.

[0019] Also within the scope of the present disclosure is a method for producing an anti-HIV antibody or a fragment thereof. The method includes: (a) obtaining the aforementioned cultured cell; (b) culturing the cell in a medium under conditions that allow for the expression of the polypeptide encoded by the vector and the assembly of the antibody or a fragment thereof; and (c) purifying the antibody or fragment from the cultured cell or the medium of the cell.

[0020] In another aspect, the present disclosure also provides a pharmaceutical composition comprising: (i) the aforementioned at least one anti-HIV antibody or an antigen-binding portion thereof, nucleic acid, or vector; and (ii) a pharmaceutically acceptable carrier.

[0021] In some embodiments, the pharmaceutical composition further comprises a second therapeutic agent. In some embodiments, the second therapeutic agent comprises an antiviral agent or one or more additional antibodies. In some embodiments, the one or more additional antibodies comprise a second anti-HIV antibody (e.g., a disclosed isolated anti-HIV bNAb) or an antigen-binding portion thereof, or a third antibody that binds to a third antigen.In some embodiments, the third antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-l), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), CD160 (NK1), immune checkpoint proteins (e.g., PD-1, PD-L1, CTLA-4), and the second epitope of gp120.

[0022] In some embodiments, the antiviral agent is selected from the group consisting of non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry inhibitors or fusion inhibitors, and integrase inhibitors.

[0023] In yet other aspects, the present disclosure provides a method for preventing or treating HIV infection or an HIV-related disease. The method includes (a) identifying a patient in need of such prevention or treatment, and (b) administering to the patient a first therapeutic agent comprising a therapeutically effective amount of at least one anti-HIV antibody or an antigen-binding portion thereof, as described above.

[0024] In some embodiments, the method further comprises administering a second therapeutic agent. In some embodiments, the second therapeutic agent comprises an antiviral agent or one or more additional antibodies. In some embodiments, the one or more additional antibodies comprise a second anti-HIV antibody (e.g., an isolated anti-HIV bNAb as disclosed) or an antigen-binding portion thereof, or a third antibody that binds to a third antigen.In some embodiments, the third antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-l), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), CD160 (NK1), immune checkpoint proteins (e.g., PD-1, PD-L1, CTLA-4), and the second epitope of gp120.

[0025] In some embodiments, the first therapeutic agent or the second therapeutic agent is administered to the patient intratumorally, intravenously, subcutaneously, intraosseously, orally, transdermally, or sublingually. In some embodiments, the first therapeutic agent is administered to the patient before, after, or simultaneously with the second therapeutic agent.

[0026] In other aspects, the disclosure further provides a kit comprising a pharmaceutically acceptable dosage unit of a pharmaceutically effective amount of at least one of the aforementioned isolated anti-HIV antibodies, or antigen-binding portion thereof.

[0027] In some embodiments, the kit further comprises a pharmaceutically acceptable dosage unit of a pharmaceutically effective amount of an anti-HIV agent. The two pharmaceutically acceptable dosage units may be in the form of a single pharmaceutically acceptable dosage unit. In some embodiments, the anti-HIV agent is selected from the group consisting of non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry inhibitors or fusion inhibitors, and integrase inhibitors.

[0028] The foregoing summary is not intended to define all aspects of the disclosure, and additional aspects are described in other sections such as the following detailed description. The entire document is intended to be related as a unified disclosure, and it should be understood that all combinations of features described herein are intended, even if the combinations are not found together in the same sentence, paragraph, or section of the document. Other features and advantages of the invention will become apparent from the following detailed description. However, it should be understood that various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description, and thus the detailed description and specific examples are provided to illustrate specific embodiments of the disclosure, but are given by way of example only. BRIEF DESCRIPTION OF THE DRAWINGS

[0029]

Figure 1

Figure 2

Figure 3

DETAILED DESCRIPTION OF THE INVENTION

[0030] Detailed Description of the Invention The present disclosure is based, at least in part, on the unexpected discovery that the disclosed variants of the anti-HIV bNAb NIH45-46 exhibited improved biophysical properties while retaining broad and potent neutralizing activity. Anti-HIV bNAb variants and antigen-binding fragments as disclosed herein constitute novel therapeutic strategies for treating and / or preventing HIV infection.

[0031] A. Novel Anti-HIV Broadly Neutralizing Antibodies a. Antibodies The present invention disclosed herein includes novel isolated anti-HIV broadly neutralizing antibodies or antigen-binding portions thereof with improved biophysical properties such as reduced polyreactivity and extended half-life.

[0032] In some embodiments, the isolated anti-HIV antibody, or antigen-binding portion thereof, comprises a heavy chain having one or more heavy chain substitutions at one or more residues selected from the group consisting of S5, S21, G54, G55, V57, T68, T70, V73, S75, F79, S82, D85, V89, Y97, and P112, with residue numbering indicated by Kabat numbering, and having a heavy chain amino acid sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 1; or a light chain having one or more light chain substitutions at one or more residues selected from the group consisting of T5, S7, T10, S12, S14, T18, I20, S22, R24, Q27, S28, S30, R59, S61, S63, W65, D68, N70, S72, S74, and S78, and having a light chain amino acid sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 2.

[0033] In some embodiments, the isolated anti-HIV antibody or antigen-binding portion thereof comprises a heavy chain polypeptide sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 1, wherein the heavy chain amino acid sequence comprises one or more heavy chain substitutions at one or more residues selected from the group consisting of S5, S21, G54, G55, V57, T68, T70, V73, S75, F79, S82, D85, V89, Y97, and P112; and a light chain amino acid sequence that is at least 75% (e.g., 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO: 2, wherein the light chain amino acid sequence comprises one or more light chain substitutions at one or more residues selected from the group consisting of T5, S7, T10, S12, S14, T18, I20, S22, R24, Q27, S28, S30, R59, S61, S63, W65, D68, N70, S72, S74, and S78.

[0034] In some embodiments, the one or more heavy chain substitutions are a substitution of S5D or S5E, or a conservative substitution of Asp or Glu at residue S5; a substitution of S21D or S21E, or a conservative substitution of Asp or Glu at residue S21; a substitution of G54W or a conservative substitution of Trp at residue G54; a substitution of G55D or G55E, or a conservative substitution of Asp or Glu at residue G55; a substitution of V57D or V57E, or a conservative substitution of Asp or Glu at residue V57; a substitution of T68D or T68E, or a conservative substitution of Asp or Glu at residue T68; a substitution of T70D or T70E, or a conservative substitution of Asp or Glu at residue T70; a substitution of V73D or V73E, or a conservative substitution of Asp or Glu at residue V73; a substitution of S75D or S75E, or a conservative substitution of Asp or Glu at residue S75; a substitution of F79D, F79E, F79Y or F79H, or a conservative substitution of Asp, Glu, Tyr or His at residue F79; a substitution of S82D or S82E, or a conservative substitution of Asp or Glu at residue S82; a substitution of D85E, or a conservative substitution of Asp at residue D85; a substitution of V89D or V89E, or a conservative substitution of Asp or Glu at residue V89; a substitution of Y97D or Y97E, or a conservative substitution of Asp or Glu at residue Y97; a substitution of P112D or P112E, or a conservative substitution of Asp or Glu at residue P112; or a combination thereof.

[0035] In some embodiments, the one or more light chain substitutions are substitutions of T5D or T5E, or conservative substitutions of Asp or Glu at residue T5; substitutions of S7D or S7E, or conservative substitutions of Asp or Glu at residue S7; substitutions of T10D or T10E, or conservative substitutions of Asp or Glu at residue T10; substitutions of S12D or S12E, or conservative substitutions of Asp or Glu at residue S12; substitutions of S14D or S14E, or conservative substitutions of Asp or Glu at residue S14; substitutions of T18D or T18E, or conservative substitutions of Asp or Glu at residue T18; substitutions of I20D or I20E, or conservative substitutions of Asp or Glu at residue I20; substitutions of S22D or S22E, or conservative substitutions of Asp or Glu at residue S22; substitutions of R24D or R24E, or conservative substitutions of Asp or Glu at residue R24; substitutions of Q27D or Q27E, or conservative substitutions of Asp or Glu at residue Q27; substitutions of S28Y or S28H, or conservative substitutions of Tyr or His at residue S28; substitutions of S30D or S30E, or conservative substitutions of Asp or Glu at residue S30; substitutions of R59D or R59E, or conservative substitutions of Asp or Glu at residue R59; substitutions of S61D or S61E, or conservative substitutions of Asp or Glu at residue S61; substitutions of S63D or S63E, or conservative substitutions of Asp or Glu at residue S63; substitutions of W65D, W65E, W65Y or W65H, or conservative substitutions of Asp, Glu, Tyr or His at residue W65; substitutions of D68E, or conservative substitutions of Asp at residue D68; substitutions of N70D or N70E, or conservative substitutions of Asp or Glu at residue N70; substitutions of S72D or S72E, or conservative substitutions of Asp or Glu at residue S72; substitutions of S74D or S74E, or conservative substitutions of Asp or Glu at residue S74; substitutions of S78D or S78E, or conservative substitutions of Asp or Glu at residue S78; or combinations thereof.

[0036] In some embodiments, the one or more heavy chain substitutions further include the substitutions M428L and N434S.

[0037] In some embodiments, the light chain amino acid sequence includes one or more light chain substitutions of (i) Q27E, S28H, and S30D; (ii) Q27E, S28H, S30D, and S74T; (iii) Q27E, S28H, S30D, S74T, M428L, and N434S; (iv) Q27E, S28Y, and S30D; (v) Q27D and S28H; (vi) S28Y; (vii) T5D, T10D, S12D, S14D, I20D, and S22D; or (viii) S61D, S63D, W65D, N70D, S72D, and S74D.

[0038] In some embodiments, the heavy chain amino acid sequence includes one or more heavy chain substitutions including G54W and T68D; and the light chain amino acid sequence includes one or more light chain substitutions of (a) Q27E, S28H, S30D, and S74T; (b) Q27E, S28H, S30D, S74T, M428L, and N434S; (c) Q27E, S28Y, and S30D; (d) Q27D and S28H; or (e) S28Y.

[0039] In some embodiments, the heavy chain amino acid sequence includes one or more heavy chain substitutions including G54W; and the light chain amino acid sequence includes one or more light chain substitutions of (a) Q27E, S28H, and S30D; (b) Q27E, S28Y, and S30D; or (c) Q27D and S28H.

[0040] In some embodiments, the heavy chain amino acid sequence includes the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3. In some embodiments, the light chain amino acid sequence includes the amino acid sequences of SEQ ID NO: 2 and SEQ ID NOs: 4-10.

[0041] In some embodiments, the heavy chain amino acid sequence comprises the amino acid sequences of SEQ ID NOs: 3 to 35. In some embodiments, the light chain amino acid sequence comprises the amino acid sequences of SEQ ID NOs: 2 and 36 to 80.

[0042] In some embodiments, the isolated anti-HIV antibody comprises CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3, each comprising the respective sequences shown in the set of CDR sequences of SEQ ID NOs: 81, 84, 87, 90, 93, and 96; SEQ ID NOs: 81, 84, 87, 99, 93, and 96; SEQ ID NOs: 81, 84, 87, 100, 93, and 96; SEQ ID NOs: 81, 84, 87, 101, 93, and 96; or SEQ ID NOs: 81, 84, 87, 102, 93, and 96; (shown in Kabat numbering); SEQ ID NOs: 82, 85, 88, 91, 94, and 97 (shown in IMGT numbering); or SEQ ID NOs: 83, 86, 89, 92, 95, and 98 (shown in Chothia numbering).

[0043] In some embodiments, the heavy chain amino acid sequence and the light chain amino acid sequence each comprise the respective amino acid sequences shown in a set of sequences selected from the group consisting of SEQ ID NOs: 3 and 36; SEQ ID NOs: 3 and 38; SEQ ID NOs: 3 and 39; SEQ ID NOs: 4 and 2; SEQ ID NOs: 4 and 38; SEQ ID NOs: 4 and 39; SEQ ID NOs: 4 and 41; and SEQ ID NOs: 34 and 37.

[0044]

Table 1-1

[0045]

Table 1-2

[0046]

Table 1-3

[0047]

Table 1-4

[0048]

Table 1-5

[0049]

Table 1-6

[0050]

Table 1-7

[0051]

Table 1-8

[0052]

Table 1-9

[0053]

Table 1-10

[0054]

Table 1-11

[0055]

Table 1-12

[0056]

Table 1-13

[0057]

Table 1-14

[0058]

Table 1-15

[0059]

Table 1-16

[0060]

Table 1-17

[0061]

Table 1-18

[0062]

Table 1-19

[0063]

Table 1-20

[0064] In some embodiments, the antibody or antigen-binding fragment thereof further comprises a mutant Fc constant region. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a chimeric antibody, a humanized antibody, or a humanized monoclonal antibody. The antibody is a single-chain antibody, a monovalent antibody, a single-chain variable fragment (scFv), an scFv-Fc fusion, a minibody, a Fab, or an F(ab’)2 fragment.

[0065] In some embodiments, the antibody or antigen-binding fragment thereof can be detectably labeled or conjugated to a toxin, therapeutic agent, polymer (e.g., polyethylene glycol (PEG)), receptor, enzyme, or receptor ligand. For example, the antibodies of the invention can bind to a toxin (e.g., tetanus toxin).

[0066] In other examples, the antibodies of the invention can bind to a detectable tag. Such antibodies can be used in diagnostic assays to determine whether an animal, such as a human, is infected with HIV-1. Examples of detectable tags include fluorescent proteins (i.e., green fluorescent protein, red fluorescent protein, yellow fluorescent protein), fluorescent markers (i.e., fluorescein isothiocyanate, rhodamine, Texas red), radiolabels (i.e., 3H, 32P, 125I), enzymes (i.e., β-galactosidase, horseradish peroxidase, β-glucuronidase, alkaline phosphatase), or affinity tags (i.e., avidin, biotin, streptavidin, Fc). Methods for conjugating antibodies to detectable tags are known in the art. Harlow et al., Antibodies: A Laboratory Manual, page 319 (Cold Spring Harbor Pub. 1988).

[0067] b. Fragment In some embodiments, the antibodies provided herein are antibody fragments. Antibody fragments include, but are not limited to, Fab, Fab’, Fab’-SH, F(ab’)2, Fv, and single-chain Fv (scFv) fragments, as well as other fragments described below, such as diabodies, triabodies, tetra-bodies, and single-domain antibodies. For a review of certain antibody fragments, see Hudson et al., Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, for example, Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994); further see WO 93 / 16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab’)2 fragments that include salvage receptor binding epitope residues and have increased in vivo half-lives, see U.S. Patent No. 5,869,046.

[0068] A diabody is an antibody fragment that has two antigen-binding sites that are bivalent or bispecific. See, for example, EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Triabodies and tetra-bodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0069] A single-domain antibody is an antibody fragment that includes all or part of the heavy-chain variable region of an antibody, or all or part of the light-chain variable region. In some embodiments, the single-domain antibody is a human single-domain antibody (see DOMANTIS, Inc., Waltham, Mass.; e.g., U.S. Patent No. 6,248,516).

[0070] Antibody fragments can be produced by a variety of techniques including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells (e.g., E. coli or phage) as described herein.

[0071] c. Chimeric and humanized antibodies In some embodiments, the antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984). In one example, a chimeric antibody includes a non-human variable region (e.g., a variable region derived from a non-human primate such as a mouse, rat, hamster, rabbit, or monkey) and a human constant region. In a further example, a chimeric antibody is a "class-switch" antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0072] In some embodiments, the chimeric antibody is a humanized antibody. Typically, non-human antibodies are humanized to reduce their immunogenicity in humans while retaining the specificity and affinity of the parental non-human antibody. Usually, a humanized antibody comprises one or more variable regions in which the HVRs, e.g., CDRs (or portions thereof) are derived from a non-human antibody and the FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody may also comprise at least a portion of a human constant region. In some embodiments, some FR residues of the humanized antibody are replaced with corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived) to, for example, restore or improve the specificity or affinity of the antibody.

[0073] Humanized antibodies and methods of making them are reviewed, for example, in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described, for example, in Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Nat’l Acad. Sci. USA 86:10029-10033 (1989); U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specific-determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"); Dall’Acqua et al., Methods 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing "guided selection" approaches to FR shuffling).

[0074] Human framework regions that can be used for humanization include framework regions selected using an "optimal" method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from consensus sequences of human antibodies of certain subgroups of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatic mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions obtained from screening of FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)), but are not limited thereto.

[0075] d. Human antibodies In some embodiments, the antibodies provided herein are human antibodies. Human antibodies can be produced using various techniques known in the art or using the techniques described herein. Human antibodies are generally described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20:450-459 (2008).

[0076] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies having human variable regions in response to an antigen challenge. Such animals typically contain all or part of a human immunoglobulin locus that replaces the endogenous immunoglobulin locus, or is present extrachromosomally, or is randomly integrated into the chromosomes of the animal. In such transgenic mice, the endogenous immunoglobulin locus is usually inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). Further, see, for example, U.S. Patent Nos. 6,075,181 and 6,150,584, which describe the XENOMOUSE technology; U.S. Patent No. 5,770,429, which describes the HUMAB technology; U.S. Patent No. 7,041,870, which describes the K-M MOUSE technology; and U.S. Provisional Patent Application No. 2007 / 0061900, which describes the VELOCIMOUSE technology. The human variable regions from intact antibodies produced by such animals can be further modified, for example, by combining them with different human constant regions.

[0077] Human antibodies can also be produced by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described. (See, e.g., Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991).) Human antibodies produced via human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006). Additional methods include, for example, those described in U.S. Patent No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines), and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (triooma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005), and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3):185-91 (2005).

[0078] Human antibodies can also be produced by isolating Fv clone variable region sequences selected from a human-derived phage display library. Such variable region sequences can then be combined with desired human constant regions. Techniques for selecting human antibodies from antibody libraries are described below.

[0079] The antibodies of the present invention can be isolated by screening a combinatorial library for antibodies having a desired activity or activities. For example, phage display libraries can be generated and various methods for screening such libraries for antibodies having desired binding properties are known in the art. Such methods are discussed, for example, in Hoogenboom et al., in Methods in Molecular Biology 178:1-37 (O’Brien et al., ed., Human Press, Totowa, N.J., 2001), and further described, for example, in McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, N.J., 2003); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004).

[0080] In certain phage display methods, repertoires of VH and VL genes are separately cloned by polymerase chain reaction (PCR), as described in Winter et al., Ann. Rev. Immunol., 12: 433-455 (1994), randomly recombined in a phage library, and then screened for antigen-binding phage. The phage typically displays an antibody fragment, either as a scFv fragment or a Fab fragment. A library from an immunogen provides high-affinity antibodies against the immunogen without the need to construct hybridomas. Alternatively, a naive repertoire can be cloned (e.g., from humans) to provide a single source of antibodies against a wide range of non-self and self antigens without immunization, as described in Griffiths et al., EMBO J, 12: 725-734 (1993). Finally, naive libraries can also be produced synthetically by cloning unrearranged V gene segments from stem cells and using PCR primers containing random sequences to encode a highly variable CDR3 region and achieve rearrangement in vitro. Patent publications describing human antibody phage libraries include, for example, U.S. Patent No. 5,750,373, and U.S. Patent Application Publications Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360. Antibodies or antibody fragments isolated from a human antibody library are considered herein to be human antibodies or human antibody fragments.

[0081] e. Variant In some embodiments, amino acid sequence variants of the antibodies provided herein are contemplated. For example, it may be desirable to improve the binding affinity of the antibody and / or other biological properties (e.g., reduction of polyreactivity, increase in half-life, etc.). Amino acid sequence variants of the antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions from, and / or insertions into, and / or substitutions of residues within the amino acid sequence of the antibody. Combinations of any of deletions, insertions, and substitutions can be made to arrive at the final construct, provided that the final construct has the desired properties, such as antigen binding.

[0082] Substitution, insertion, and deletion variants In some embodiments, antibody variants having one or more amino acid substitutions are provided. Sites for substitution mutagenesis include HVRs and FRs. Conservative substitutions are defined herein. Amino acid substitutions can be introduced into the antibody of interest and the product is screened for the desired activity, such as retention / improvement of antigen binding, reduction of immunogenicity, or improvement of antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC).

[0083] Accordingly, the antibodies of the present invention may include one or more conservative modifications of the CDRs, heavy chain variable regions, or light variable regions described herein. Conservative modifications or functional equivalents of the peptides, polypeptides, or proteins disclosed in the present invention refer to polypeptide derivatives of peptides, polypeptides, or proteins, such as proteins having one or more point mutations, insertions, deletions, truncations, fusion proteins, or combinations thereof. It substantially retains the activity of the parent peptide, polypeptide, or protein (such as those disclosed in the present invention). Usually, the conservative modification or equivalent function is at least 60% identical to the parent (for example, any numerical value from 60% to 100%, such as 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, and 99%). Accordingly, heavy chain variable regions or light variable regions having one or more point mutations, insertions, deletions, truncations, fusion proteins, or combinations thereof, and antibodies having mutant regions are within the scope of the present invention.

[0084] As used herein, percent homology between two amino acid sequences is equivalent to percent identity between the two sequences. The percentage of identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap (i.e., % homology = number of identical positions / total number of positions × 100). As illustrated in the following non-limiting examples, comparison of sequences and determination of the percent identity between two sequences can be accomplished using mathematical algorithms.

[0085] The percent identity between two amino acid sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci., 4:11-17 (1988)), incorporated into the ALIGN program (version 2.0), using the PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Further, the percent identity between two amino acid sequences can be determined using the algorithm of Needleman and Wunsch (J. Mol. Biol. 48:444-453 (1970)), incorporated into the GAP program of the GCG software package (available at www.gcg.com), using either the Blossum 62 matrix or the PAM250 matrix, and gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6.

[0086] Additionally or alternatively, the protein sequences of the invention can further be used, for example, as a "query sequence" to perform a search against a public database in order to identify related sequences. Such studies can be performed using the XBLAST program (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. A BLAST protein search can be performed using the XBLAST program, score = 50, wordlength = 3, to obtain amino acid sequences homologous to the antibody molecules of the invention. To obtain gapped alignments for comparison purposes, gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402. When utilizing the BLAST and gapped BLAST programs, the default parameters of each program (e.g., XBLAST and NBLAST) can be used. (See www.ncbi.nlm.nih.gov).

[0087] As used herein, the terms "conservative modification" or "conservative substitution" refer to amino acid modifications that do not significantly affect or alter the binding properties of an antibody comprising an amino acid sequence. Such conservative modifications include amino acid substitutions, additions, and deletions. The modifications can be introduced into the antibodies of the invention by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are those in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are defined in the art. These families include (i) amino acids having basic side chains (e.g., lysine, arginine, histidine), (ii) acidic side chains (e.g., aspartic acid, glutamic acid), (iii) uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), (iv) nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), (v) beta-branched side chains (e.g., threonine, valine, isoleucine), and (vi) aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0088] Alternatively, as described in Lehninger [Biochemistry, Second Edition; Worth Publishers, Inc. NY, N.Y. (1975), pp. 71-77], conservative amino acids can be grouped as follows:

[0089] [Table 2]

[0090] As yet another alternative, exemplary conservative substitutions are shown below:

[0091] [Table 3]

[0092] Conservative substitutions of existing substitutions refer to conservative substitutions of the substituted residues. For example, a conservative substitution of Trp at residue G54 (or position 54) refers to a conservative substitution of Trp at position 54, i.e., Tyr (Y). Similarly, a conservative substitution of Tyr at residue S30 (or position 30) refers to a conservative substitution of Tyr at position 30, i.e., Trp, Phe, Thr, Ser.

[0093] Non-conservative substitutions necessarily involve an exchange from one member of these classes to another.

[0094] Exemplary substitution variants are affinity matured antibodies, which can be conveniently produced using, for example, phage display-based affinity maturation techniques as described in Hoogenboom et al., in Methods in Molecular Biology 178:1-37 (O’Brien et al., ed., Human Press, Totowa, N.J., (2001)). Amino acid sequence insertions include amino-terminal and / or carboxyl-terminal fusions of lengths up to polypeptides containing from 1 residue to over 100 residues, as well as in-sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies having an N-terminal methionyl residue. Other insertion variants of antibody molecules include fusions of antibodies to enzymes (e.g., in the case of ADEPT) or polypeptides that increase the serum half-life of the antibody, at the N-terminal or C-terminal of the antibody.

[0095] Glycosylation variants In some embodiments, the antibodies provided herein are modified to increase or decrease the degree to which the antibody is glycosylated. Addition or deletion of glycosylation sites to the antibody can be conveniently achieved by altering the amino acid sequence such that one or more glycosylation sites are created or removed.

[0096] For example, non-glycosylated antibodies can be produced (i.e., the antibodies lack glycosylation). Glycosylation can be modified, for example, to enhance the affinity of the antibody for the antigen. Such carbohydrate modifications can be achieved, for example, by altering one or more sites of glycosylation within the antibody sequence. For example, one or more amino acid substitutions can be made that result in the elimination of glycosylation sites in one or more variable region frameworks, thereby eliminating glycosylation at that site. Such glycosylation can enhance the affinity of the antibody for the antigen. Such an approach is described in more detail in U.S. Patents Nos. 5,714,350 and 6,350,861 by Co et al. Glycosylation of the constant region at N297 can be inhibited by mutating the N297 residue to another residue, for example, N297A, and / or by mutating an adjacent amino acid, such as 298, thereby reducing glycosylation at N297.

[0097] Additionally or alternatively, antibodies with altered glycosylation types can be produced, such as hypofucosylated antibodies with a reduced amount of fucosyl residues or antibodies with an increased bisecting GlcNac structure. Such altered glycosylation patterns have been demonstrated to enhance the ADCC ability of antibodies. Such carbohydrate modifications can be achieved, for example, by expressing the antibody in a host cell having a modified glycosylation mechanism. Cells with altered glycosylation mechanisms have been described in the art and can be used as host cells for expressing the recombinant antibodies described herein and thereby producing antibodies with altered glycosylation. For example, EP 1,176,195 by Hanai et al. describes that cell lines having a functionally disrupted FUT8 gene encode fucosyltransferase and that antibodies expressed in such cell lines exhibit hypofucosylation. PCT application WO 03 / 035835 by Presta describes that Led3 cells, which are mutants of Chinese hamster ovary cell lines, also cause hypofucosylation of antibodies expressed in their host cells due to a reduced ability to bind fucose to the Asn(297)-linked carbohydrate (see also Shields, R.L. et al. (2002) J. Biol. Chem. 277:26733-26740). PCT application WO 99 / 54342 by Umana et al. describes that cell lines (e.g., beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII)) are designed to express a glycoprotein-modifying glycosyltransferase, and that antibodies expressed in the designed cell lines exhibit an increase in the bisecting GlcNac structure, resulting in an increase in the ADCC activity of the antibodies (see also Umana et al. (1999) Nat. Biotech. 17: 176-180).

[0098] Variant of the Fc region The variable regions of the antibodies described herein can be linked (e.g., covalently or by fusion) to an Fc, such as an IgG1 Fc, IgG2 Fc, IgG3 Fc, or IgG4 Fc, which can be of any allotype or isoallotype, e.g., for IgG1: Glm, Glml(a), Glm2(x), Glm3(f), Glml7(z); for IgG2: G2m, G2m23(n); for IgG3: G3m, G3m21(gl), G3m28(g5), G3ml l(b0), G3m5(bl), G3ml3(b3), G3ml4(b4), G3ml0(b5), G3ml5(s), G3ml6(t), G3m6(c3), G3m24(c5), G3m26(u), G3m27(v); for K: Km, Kml, Km2, Km3 (see, e.g., Jefferies et al. (2009) mAbs 1: 1). In some embodiments, the antibody variable regions described herein are linked to an Fc that binds to one or more activating Fc receptors (FcγI, Fcγlla, or FcγIIIa), thereby stimulating ADCC and causing T cell depletion. In some embodiments, the antibody variable regions described herein are linked to an Fc that causes depletion.

[0099] In some embodiments, the antibody variable regions described herein can be linked to an Fc containing one or more modifications to alter one or more functional properties of the antibody, such as typically serum half-life, complement binding, Fc receptor binding, and / or antibody-dependent cell-mediated cytotoxicity. Further, the antibodies described herein can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or modified to change one or more functional properties of the antibody, such as by changing its glycosylation. The numbering of the residues in the Fc region is according to the Kabat EU index numbering.

[0100] The Fc region includes domains derived from the constant region of an immunoglobulin, preferably a human immunoglobulin, and includes fragments, analogs, variants, mutants or derivatives of the constant region. Suitable immunoglobulins include IgG1, IgG2, IgG3, IgG4, and other classes such as IgA, IgD, IgE and IgM. The constant region of the immunoglobulin is defined as a natural or synthetically produced polypeptide homologous to the C-terminal region of the immunoglobulin, and may include the CH1 region, hinge, CH2 region, CH3 region, or CH4 region individually or in combination. In some embodiments, the antibodies of the invention have an Fc region other than wild-type IgA1. The antibody may have an Fc region from IgG (e.g., IgG1, IgG2, IgG3, and IgG4) or other classes such as IgA2, IgD, IgE, and IgM. The Fc may be a variant of IgA1.

[0101] The constant region of the immunoglobulin is involved in many important antibody functions such as Fc receptor (FcR) binding and complement binding. There are five major classes of heavy chain constant regions, classified as IgA, IgG, IgD, IgE, IgM, each with characteristic effector functions specified by the isotype. For example, IgG is divided into four subclasses known as IgG1, IgG2, IgG3, and IgG4.

[0102] Ig molecules interact with multiple classes of cell receptors. For example, IgG molecules interact with three classes of Fcγ receptors (FcγR) specific for antibodies of the IgG class, namely FcγRI, FcγRII, and FcγRIIL. It has been reported that important sequences for IgG to bind to the FcγR receptor are located in the CH2 region and CH3 region. The serum half-life of an antibody is affected by its ability to bind to FcR.

[0103] In some embodiments, the Fc region is a variant Fc region, such as an Fc sequence modified (e.g., by amino acid substitution, deletion, and / or insertion) with respect to a parental Fc sequence (e.g., an unmodified Fc polypeptide that is subsequently modified to generate a variant) to provide desired structural features and / or biological activities. For example, the Fc region can be modified to generate Fc variants that (a) have increased or decreased ADCC, (b) have increased or decreased CDC, (c) have increased or decreased affinity for Clq, and / or (d) have increased or decreased affinity for Fc receptors compared to the parental Fc. Such Fc region variants will typically include at least one amino acid modification in the Fc region. It is particularly desirable to combine amino acid modifications. For example, the variant Fc region can include therein, for example, 2, 3, 4, 5, etc. substitutions at specific Fc region positions identified herein.

[0104] The variant Fc region may also include sequence modifications in which the amino acids involved in disulfide bond formation are removed or replaced with other amino acids. Such removal may avoid reaction with other cysteine-containing proteins present in the host cell used to produce the antibodies described herein. Even when cysteine residues are removed, the single-chain Fc region can form a non-covalently bound dimeric Fc region. In some embodiments, the Fc region may be modified to enhance compatibility with the selected host cell. For example, the PA sequence near the N-terminus of a typical native Fc region, which can be recognized by digestive enzymes of E. coli such as prolidase, may be removed. In other embodiments, one or more glycosylation sites within the Fc region may be removed. Residues that are normally glycosylated (e.g., asparagine) can cause cell lysis reactions. Such residues may be removed or replaced with non-glycosylated residues (e.g., alanine). In other embodiments, sites involved in interaction with complement, such as the Clq binding site, may be removed from the Fc region. For example, the EKK sequence of human IgG1 may be removed or replaced. In some embodiments, sites that affect binding to Fc receptors, preferably sites other than the salvage receptor binding site, may be removed. In other embodiments, the Fc region may be modified to remove the ADCC site. The ADCC site is known in the art; see, for example, Molec. Immunol. 29 (5): 633-9 (1992) for the ADCC site of IgG1. Specific examples of Fc regions are disclosed, for example, in WO 97 / 34631 and WO 96 / 32478.

[0105] In one embodiment, the hinge region of the Fc is modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased. This approach is further described in U.S. Patent No. 5,677,425 by Bodmer et al. The number of cysteine residues in the hinge region of the Fc is altered, e.g., to facilitate the assembly of the light and heavy chains, or to increase or decrease the stability of the antibody. In one embodiment, the Fc hinge region of the antibody is mutated to shorten the biological half-life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 region interface region of the Fc hinge fragment, weakening the binding of the antibody to staphylococcal protein A (SpA) compared to the binding of the native Fc hinge region to SpA. This approach is further described in detail in U.S. Patent No. 6,165,745 by Ward et al.

[0106] In still other embodiments, the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to change the effector function of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320, and 322 may be replaced with different amino acid residues such that the antibody has an altered affinity for an effector ligand while retaining the antigen-binding ability of the parental antibody. The effector ligand whose affinity is changed can be, for example, an Fc receptor or the CI component of complement. This approach is further described in detail in U.S. Patents Nos. 5,624,821 and 5,648,260 by Winter et al.

[0107] In other examples, one or more amino acids selected from amino acid residues 329, 331, and 322 may be replaced with different amino acid residues such that the antibody alters Clq binding or reduces or abrogates CDC. This approach is further described in detail in U.S. Patent No. 6,194,551 by Idusogie et al.

[0108] In other examples, one or more amino acid residues within positions 231 and 239 of the amino acid are altered, thereby altering the ability of the antibody to fix complement. This approach is described in detail in PCT application WO 94 / 29351 by Bodmer et al.

[0109] In still other examples, the Fc region can be modified to increase ADCC and / or increase affinity for the Fcγ receptor by modifying one or more amino acids at the following positions: 234, 235, 236, 238, 239, 240, 241, 243, 244, 245, 247, 248, 249, 252, 254, 255, 256, 258, 262, 263, 264, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 299, 301, 303, 305, 307, 309, 312, 313, 315, 320, 322, 324, 325, 326, 327, 329, 330, 331, 332, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, 388, 389, 398, 414, 416, 419, 430, 433, 434, 435, 436, 437, 438 or 439. Exemplary substitutions include 236A, 239D, 239E, 268D, 267E, 268E, 268F, 324T, 332D, and 332E. Exemplary mutations include 239D / 332E, 236A / 332E, 236A / 239D / 332E, 268F / 324T, 267E / 268F, 267E / 324T, and 267E / 268F / 324T. Other modifications for enhancing FcγR and complement interactions include, but are not limited to, substitutions of 298A, 333A, 334A, 326A, 247I, 339D, 339Q, 280H, 290S, 298D, 298V, 243L, 292P, 300L, 396L, 305I, and 396L. These and other modifications are reviewed in Strohl, 2009, Current Opinion in Biotechnology 20:685-691.

[0110] Fc modifications that increase binding to the Fcγ receptor include one or more amino acid modifications at positions 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 279, 280, 283, 285, 298, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 312, 315, 324, 327, 329, 330, 335, 337, 3338, 340, 360, 373, 376, 379, 382, 388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438 or 439 of the Fc region, where the numbering of the residues of the Fc region is the EU index numbering as in the case of Kabat (WO00 / 42072).

[0111] Other Fc modifications that can be made to Fc reduce or eliminate binding to FcγR and / or complement proteins, thereby reducing or eliminating Fc-mediated effector functions such as ADCC, antibody-dependent cell-mediated phagocytosis (ADCP), and CDC. Exemplary modifications include, but are not limited to, substitutions, insertions, and deletions at positions 234, 235, 236, 237, 267, 269, 325, and 328, numbered according to the EU index. Exemplary substitutions include, but are not limited to, 234G, 235G, 236R, 237K, 267R, 269R, 325L, and 328R. The Fc variant can include 236R / 328R. Other modifications for reducing FcγR and complement interactions include substitutions of 297A, 234A, 235A, 237A, 318A, 228P, 236E, 268Q, 309L, 330S, 331S, 220S, 226S, 229S, 238S, 233P, and 234V, and removal of glycosylation at position 297 by mutagenic or enzymatic means or by production in organisms such as bacteria that do not glycosylate proteins. These and other modifications are reviewed in Strohl, 2009, Current Opinion in Biotechnology 20:685-691.

[0112] Optionally, the Fc region may contain amino acid residues that do not naturally occur at additional and / or alternative positions known to those of skill in the art (see, for example, U.S. Patent Applications Nos. 5,624,821; 6,277,375; 6,737,056; 6,194,551; 7,317,091; 8,101,720; WO00 / 42072; WO01 / 58957; WO02 / 06919; WO04 / 016750; WO04 / 029207; WO04 / 035752; WO04 / 074455; WO04 / 099249; WO04 / 063351; WO05 / 070963; WO05 / 040217, WO05 / 092925, and WO06 / 020114).

[0113] Fc variants that enhance the affinity for the inhibitory receptor FcγRIIb can also be used. Such variants can provide, for example, Fc fusion proteins having immunomodulatory activity associated with FcγRIIb cells, including B cells and monocytes. In some embodiments, the Fc variant provides selectively enhanced affinity for FcγRIIb compared to one or more activating receptors. Modifications for altering binding to FcγRIIb include one or more modifications at positions selected from the group consisting of 234, 235, 236, 237, 239, 266, 267, 268, 325, 326, 327, 328, and 332 according to the EU index. Exemplary substitutions for enhancing FcγRIIb affinity include, but are not limited to, 234D, 234E, 234F, 234W, 235D, 235F, 235R, 235Y, 236D, 236N, 237D, 237N, 239D, 239E, 266M, 267D, 267E, 268D, 268E, 327D, 327E, 328F, 328W, 328Y, and 332E. Exemplary substitutions include 235Y, 236D, 239D, 266M, 267E, 268D, 268E, 328F, 328W, and 328Y. Other Fc variants for enhancing binding to FcγRllb include 235Y / 267E, 236D / 267E, 239D / 268D, 239D / 267E, 267E / 268D, 267E / 268E, and 267E / 328F.

[0114] The affinity and binding characteristics of the Fc region for the ligand can be determined by various in vitro assay methods (biochemical or immunology-based assays) known in the art, including but not limited to equilibrium methods (e.g., ELISA, or radioimmunoassay), or kinetics (e.g., BIACORE analysis), as well as other methods such as indirect binding assays, competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis, and chromatography (e.g., gel filtration). In these and other methods, one or more labels of the component being tested can be utilized, and / or various detection methods can be employed, including but not limited to chromogenic, fluorescent, luminescent, or isotopic labels. A detailed description of binding affinity and kinetics can be found in Paul, W. E., ed., Fundamental Immunology, 4th Ed., Lippincott-Raven, Philadelphia (1999), which focuses on the interaction of antibodies and immunogens.

[0115] In some embodiments, the antibody may be modified to extend its biological half-life. Various approaches are possible. For example, this can be accomplished by increasing the binding affinity of the Fc region for FcRn. For example, as described in U.S. Patent No. 6,277,375, one or more of the following residues can be mutated: 252, 254, 256, 433, 435, 436. Specific exemplary substitutions include one or more of the following: T252L, T254S, and / or T256F. Alternatively, to extend the biological half-life, as described in U.S. Patents Nos. 5,869,046 and 6,121,022 by Presta et al., the antibody can be modified within the CH1 or CL region to include a salvage receptor binding epitope derived from two loops of the CH2 region of the IgG Fc region. Other exemplary mutations that increase binding to FcRn and / or improve pharmacokinetic properties include substitutions at positions 259, 308, 428, and 434, such as 259I, 308F, 428L, 428M, 434S, 434H, 434F, 434Y, and 434M.Other variants that increase Fc binding to FcRn include 250E, 250Q, 428L, 428F, 250Q / 428L (Hinton et al., 2004, J. Biol. Chem. 279(8): 6213-6216, Hinton et al. 2006 Journal of Immunology 176:346-356), 256A, 272A, 286A, 305A, 307A, 307Q, 311A, 312A, 376A, 378Q, 380A, 382A, 434A (Shields et al, Journal of Biological Chemistry, 2001, 276(9):6591-6604), 252F, 252T, 252Y, 252W, 254T, 256S, 256R, 256Q, 256E, 256D, 256T, 309P, 311S, 433R, 433S, 433I, 433P, 433Q, 434H, 434F, 434Y, 252Y / 254T / 256E, 433K / 434F / 436H, 308T / 309P / 311S (Dall Acqua et al. Journal of Immunology, 2002, 169:5171-5180, Dall’Acqua et al., 2006, Journal of Biological Chemistry 281:23514-23524). Other modifications for modulating FcRn binding are described in Yeung et al., 2010, J Immunol, 182:7663-7671. In some embodiments, hybrid IgG isotypes having specific biological properties may be used. For example, an IgG1 / IgG3 hybrid variant can be constructed by substituting the IgG1 positions in the CH2 and / or CH3 regions with amino acids from IgG3 at positions where the two isotypes differ. Thus, hybrid variant IgG antibodies can be constructed that include one or more substitutions, such as 274Q, 276K, 300F, 339T, 356E, 358M, 384S, 392N, 397M, 422I, 435R, and 436F.In other embodiments described herein, the IgG1 / IgG2 hybrid variant can be constructed by substituting the IgG2 positions in the CH2 region and / or CH3 region with amino acids from IgG1 at positions where the two isotypes differ. Thus, the hybrid variant IgG antibody can be constructed with one or more substitutions, for example, one or more of the following amino acid substitutions: 233E, 234L, 235L, 236G (which refers to the insertion of glycine at position 236), and 321h.

[0116] Furthermore, the binding sites of human IgG1 for FcγR1, FcγRII, FcγRIII, and FcRn have been mapped, and variants with improved binding have been described (see Shields, R.L. et al. (2001) J. Biol. Chem. 276:6591-6604). Specific mutations at positions 256, 290, 298, 333, 334, and 339 have been shown to improve binding to FcγRIII. Furthermore, the following combination mutations have been shown to improve FcγRIII binding: T256A / S298A, S298A / E333A, S298A / K224A, and S298A / E333A / K334A, which have been shown to exhibit enhanced FcγRIIIa binding and ADCC activity (Shields et al., 2001). Other IgG1 variants with strongly enhanced binding to FcγRIIIa have been identified, including variants with the S239D / I332E and S239D / I332E / A330L mutations, which exhibit the greatest increase in affinity for FcγRIIIa, a decrease in FcγRIIb binding, and potent cytotoxic activity in cynomolgus monkeys (Lazar et al., 2006). Introduction of triple mutations such as the S239D / I332E mutation into antibodies such as alemtuzumab (CD52-specific), trastuzumab (HER2 / neu-specific), rituximab (CD20-specific), and cetuximab (EGFR-specific), which are converted to significantly enhanced ADCC activity in vitro, has been shown to enhance the ability to deplete monkey B cells (Lazar et al., 2006). Furthermore, IgG1 variants containing the L235V, F243L, R292P, Y300L, and P396L mutations, which exhibited enhanced binding to FcγRIIIa and concomitantly enhanced ADCC activity, have been identified in transgenic mice expressing human FcγRIIIa in models of B cell malignancies and breast cancer (Stavenhagen et al., 2007; Nordstrom et al., 2011).Other Fc mutations that can be used include the following: S298A / E333A / L334A, S239D / I332E, S239D / I332E / A330L, L235V / F243L / R292P / Y300L / P396L, and M428L / N434S.

[0117] In some embodiments, an Fc with reduced binding to FcγR is selected. Exemplary Fcs, such as an IgG1 Fc with reduced FcγR binding, contain the following three amino acid substitutions: L234A, L235E, and G237A.

[0118] In some embodiments, an Fc with reduced complement binding is selected. An exemplary Fc with reduced complement binding, such as an IgG1 Fc, has the following two amino acid substitutions: A330S and P331S.

[0119] In some embodiments, an Fc having substantially no effector function is selected, i.e., having reduced binding to FcγR and reduced complement binding. An exemplary effectorless Fc, such as an IgG1 Fc, contains the following five mutations: L234A, L235E, G237A, A330S, and P331S.

[0120] When using the IgG4 constant region, it is generally preferred to include the substitution S228P that mimics the hinge sequence of IgG1 and thereby stabilizes the IgG4 molecule.

[0121] Fc mutations that extend serum half-life In some embodiments, the Fc region or Fc domain of the anti-HIV gp120-directed antibody comprises amino acid modifications that promote an extension of the serum half-life of the anti-binding molecule. Mutations that extend the half-life of antibodies have been reported. In one embodiment, the Fc region or Fc domain of one or both of the heavy chain targeting CD3 and the heavy chain targeting the HIV antigen comprises a substitution of methionine to tyrosine at position 252 (EU numbering), a substitution of serine to threonine at position 254 (EU numbering), and a substitution of threonine to glutamic acid at position 256 (EU numbering). See, for example, U.S. Patent No. 7,658,921. This type of variant, called the "YTE variant," exhibits a four-fold longer half-life compared to the wild-type version of the same antibody (Dall’Acqua, et al., J Biol Chem, 281: 23514-24 (2006); Robbie, et al., Antimicrob Agents Chemotherap., 57(12):6147-6153 (2013)). In certain embodiments, the Fc region or Fc domain of one or both of the heavy chain targeting CD3 and the heavy chain targeting the HIV antigen 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 (EU numbering). Alternatively, the M428L and N434S ("LS") substitutions may extend the pharmacokinetic half-life of the multispecific antigen-binding molecule. In other embodiments, the Fc region or Fc domain of one or both of the heavy chain targeting CD3 and the heavy chain targeting the HIV antigen comprises the M428L and N434S (EU numbering) substitutions. In other embodiments, the Fc region or Fc domain of one or both of the heavy chain targeting CD3 and the heavy chain targeting the HIV antigen comprises the T250Q and M428L (EU numbering) mutations. In other embodiments, the Fc region or Fc domain of one or both of the heavy chain targeting CD3 and the heavy chain targeting the HIV antigen comprises the H433K and N434F (EU numbering) mutations.

[0122] Fc mutations that enhance effector activity In some embodiments, the Fc region or Fc domain of the anti-HIV gp120-directed antibody comprises amino acid modifications that increase post-translational and / or effector activity, such as improved FcγIIIa binding and increased antibody-dependent cellular cytotoxicity (ADCC). In some embodiments, the Fc region or Fc domain of the anti-HIV gp120-directed antibody comprises DE modifications (i.e., S239D and I332E according to EU numbering) in the Fc region. In some embodiments, the Fc region or Fc domain of the anti-HIV gp120-directed antibody comprises DEL modifications (i.e., S239D, I332E, and A330L according to EU numbering) in the Fc region. In some embodiments, the Fc region or Fc domain of the anti-HIV gp120-directed antibody comprises DEA modifications (i.e., S239D, I332E, and G236A according to EU numbering) in the Fc region. In some embodiments, the Fc region or Fc domain of the anti-HIV gp120-directed antibody comprises DEAL modifications (i.e., S239D, I332E, G236A, and A330L according to EU numbering) in the Fc region. See, for example, U.S. Patent Nos. 7,317,091; 7,662,925; 8,039,592; 8,093,357; 8,093,359; 8,383,109; 8,388,955; 8,735,545; 8,858,937; 8,937,158; 9,040,041; 9,353,187; 10,184,000; and 10,584,176. Additional amino acid modifications that enhance effector activity, such as improved FcγIIIa binding and increased ADCC, include, but are not limited to, F243L / R292P / Y300L / V305I / P396L; S298A / E333A / K334A; or L234Y / L235Q / G236W / S239M / H268D / D270E / S298A on the first Fc region, and D270E / K326D / A330M / K334E on the second Fc region (EU numbering).Amino acid mutations that increase C1q binding and complement-dependent cytotoxicity (CDC) include, but are not limited to, S267E / H268F / S324T or K326W / E333S (EU numbering). Mutations in the Fc region that enhance effector activity are reviewed, for example, in Wang, et al., Protein Cell (2018) 9(1): 63-73; and Saunders, Front Immunol. (2019) 10:1296.

[0123] In other embodiments, the anti-HIV gp120-directed antibody or antigen-binding fragment thereof has modified glycosylation, which can be introduced, for example, post-translationally or by genetic engineering. In some embodiments, the anti-HIV gp120-directed antibody or antigen-binding fragment thereof is afucosylated, for example, at glycosylation sites present in the antibody or antigen-binding fragment thereof. The most approved monoclonal antibodies are of the IgG1 isotype and have two N-linked branched complex oligosaccharides attached to the Fc region. The Fc region exerts the effector function of ADCC through interaction with leukocyte receptors of the FcγR family. Defucosylated monoclonal antibodies are monoclonal antibodies designed such that the oligosaccharides in the Fc region of the antibody do not have fucose sugar units.

[0124] f. Multivalent antibodies In one embodiment, the antibodies of the present invention can be monovalent or multivalent (e.g., divalent, trivalent, etc.). As used herein, the term "valence" refers to the number of potential target binding sites associated with an antibody. Each target binding site specifically binds to one target molecule, or to a specific position or locus on a target molecule. When the antibody is monovalent, each binding site of the molecule specifically binds to a single antigenic position or epitope. When the antibody contains multiple target binding sites (multivalent), each target binding site can specifically bind to the same or different molecules (e.g., can bind to different ligands or different antigens, or to different epitopes or positions on the same antigen). See, for example, U.S.P.N. 2009 / 0130105. In each case, at least one of the binding sites contains an epitope, motif, or domain associated with a DLL3 isoform.

[0125] In one embodiment, the antibody is a bispecific antibody in which the two chains have different specificities, as described in Millstein et al., 1983, Nature, 305:537-539. Other embodiments include antibodies having additional specificities, such as trispecific antibodies. Other more sophisticated and compatible multispecific constructs and methods for their production are described in U.S.P.N. 2009 / 0155255, and WO 94 / 04690; Suresh et al., 1986, Methods in Enzymology, 121:210; and WO96 / 27011.

[0126] As noted above, multivalent antibodies can immunospecifically bind to different epitopes of a target molecule of interest, or can immunospecifically bind to both a target molecule and a heterologous epitope, such as a heterologous polypeptide or a solid support material. In some embodiments, the multivalent antibody can include a bispecific antibody or a trispecific antibody.

[0127] A bispecific antibody is an antibody that has binding specificities for at least two different epitopes. Exemplary bispecific antibodies can bind to two different epitopes of a single antigen. Other such antibodies can combine a first antigen-binding site with a binding site for a second antigen. Alternatively, to direct a cell defense mechanism to concentrate and localize on infected cells, an anti-HIV arm can be combined with an arm that binds to a trigger molecule on a leukocyte such as a T-cell receptor molecule (e.g., CD3), or an Fc receptor of IgG (FcγR) such as FcγRI (CD64), FcγRII (CD32), and FcγRIII (CD16). Bispecific antibodies can be used to localize a cytotoxic agent to infected cells. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments (e.g., F(ab’)2 bispecific antibodies). For example, WO96 / 16673 describes a bispecific anti-ErbB2 / anti-FcγRIII antibody, and U.S. Patent No. 5,837,234 discloses a bispecific anti-ErbB2 / anti-FcγRI antibody. For example, a bispecific anti-ErbB2 / Fc alpha antibody has been reported in WO98 / 02463; and a bispecific anti-ErbB2 / anti-CD3 antibody is taught in U.S. Patent No. 5,821,337. Further, see, for example, Mouquet et al., Polyreactivity Increases The Apparent Affinity Of Anti-HIV Antibodies By Heteroligation. NATURE. 467, 591-5 (2010).

[0128] Bispecific antibodies also include cross-linked antibodies or "heteroconjugate" antibodies. For example, one of the heteroconjugate antibodies can bind to avidin and the other can bind to biotin. Such antibodies have been proposed, for example, for targeting immune system cells to unwanted cells (U.S. Patent No. 4,676,980) and for the treatment of HIV infection (WO 91 / 00360, WO 92 / 200373, and EP 03089). Heteroconjugate antibodies can be produced using any convenient cross-linking method. Suitable cross-linking agents are known in the art and are disclosed in U.S. Patent No. 4,676,980 along with many cross-linking techniques.

[0129] Methods for producing bispecific antibodies are known in the art. Conventional production of full-length bispecific antibodies is based on the co-expression of two immunoglobulin heavy chain-light chain pairs, where the two chains have different specificities (see, for example, Millstein et al., Nature, 305:537-539 (1983)). Similar procedures are disclosed, for example, in WO 93 / 08829, Traunecker et al., EMBO J., 10:3655-3659 (1991), and further, see Mouquet et al., Polyreactivity Increases The Apparent Affinity Of Anti-HIV Antibodies By Heteroligation. NATURE. 467, 591-5 (2010).

[0130] Alternatively, an antibody variable region having a desired binding specificity (antibody-antigen binding site) is fused to an immunoglobulin constant region sequence. The fusion is with an Ig heavy chain constant region comprising at least a part of the hinge region, CH2 region, and CH3 region. According to some embodiments, a first heavy chain constant region (CH1) containing a site necessary for light chain binding is present in at least one of the fusions. DNA encoding the immunoglobulin heavy chain fusion and, optionally, the immunoglobulin light chain is inserted into separate expression vectors and co-transfected into a suitable host cell. If the unequal ratio of the three polypeptide chains used in the construction provides an optimal yield of the desired bispecific antibody, this provides greater flexibility in adjusting the mutual ratio of the three polypeptide fragments in the embodiments. However, if expression of at least two polypeptide chains at equal ratios results in a high yield, or if the ratio does not significantly affect the yield of the desired chain combination, the coding sequences of two or all three polypeptide chains can be inserted into a single expression vector.

[0131] Techniques for producing bispecific antibodies from antibody fragments are also described in the above literature. For example, bispecific antibodies can be prepared using chemical linkages. For example, Brennan et al., Science, 229: 81 (1985) describes a procedure in which intact antibodies are proteolytically cleaved to produce F(ab’)2 fragments. These fragments are reduced in the presence of sodium arsenite, a dithiol complexing agent, which stabilizes adjacent dithiols and inhibits intermolecular disulfide formation. The resulting Fab’ fragments are then converted to thionitrobenzoic acid (TNB) derivatives. One of the Fab’-TNB derivatives is then reconverted to Fab’-thiol by reduction with mercaptoethylamine and mixed with an equimolar amount of another Fab’-TNB derivative to form a bispecific antibody. The produced bispecific antibody can be used as an agent for the selective immobilization of enzymes.

[0132] Other modifications of the antibody are contemplated herein. For example, the antibody may be linked to one of various non-proteinaceous polymers such as polyethylene glycol, polypropylene glycol, polyoxyalkylene, or a copolymer of polyethylene glycol and polypropylene glycol. The antibody may also be encapsulated, for example, in microcapsules prepared by coacervation techniques or interfacial polymerization (e.g., hydroxy methylcellulose or gelatin-microcapsules, respectively, and poly-(methylmethacrylate) microcapsules), colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or macroemulsions. Such techniques are disclosed, for example, in Remington’s Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980).

[0133] In some embodiments, the bispecific antibody of the invention comprises a first antigen-binding arm that binds to a first antigen (e.g., gp120) and a second antigen-binding arm that binds to a second antigen, wherein the first antigen and the second antigen are different.

[0134] In some embodiments, the first antigen-binding arm comprises: (a) VH CDR1-3 and VL CDR1-3 as set forth in SEQ ID NOs: 11-16, respectively; or (b) HC and LC as set forth in SEQ ID NO: 3 and SEQ ID NO: 2; SEQ ID NO: 3 and SEQ ID NO: 6; SEQ ID NO: 3 and SEQ ID NO: 7; SEQ ID NO: 3 and SEQ ID NO: 8; SEQ ID NO: 3 and SEQ ID NO: 9; SEQ ID NO: 3 and SEQ ID NO: 10; SEQ ID NO: 1 and SEQ ID NO: 5; SEQ ID NO: 1 and SEQ ID NO: 8; or SEQ ID NO: 1 and SEQ ID NO: 9, respectively.

[0135] In some embodiments, the second antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-l), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), CD160 (NK1), immune checkpoint proteins (e.g., PD-1, PD-L1, CTLA-4), and the second epitope of gp120.

[0136] g. Antibody derivatives The antibodies provided herein are known in the art and can be further modified to include additional non-proteinaceous moieties that are readily available. Suitable moieties for derivatizing the antibodies include, but are not limited to, water-soluble polymers.

[0137] Non-limiting examples of water-soluble polymers include PEG, copolymers of ethylene glycol / propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acids (either homopolymers or random copolymers), dextran or poly(n-vinyl pyrrolidone) polyethylene glycol, propylene glycol homopolymer, and polypropylene oxide / ethylene oxide copolymer, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof, but are not limited thereto. Polyethylene glycol propionaldehyde has advantages in production because it is stable in water. The polymers can be of any molecular weight and can be branched or unbranched. The number of polymers attached to the antibody can vary, and when multiple polymers are attached, they can be the same molecule or different molecules. Usually, the number and / or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the specific properties or functions of the antibody to be improved, whether the antibody derivative is to be used for treatment under defined conditions, etc.

[0138] In other embodiments, conjugates of antibodies and non-proteinaceous moieties are provided that can be selectively heated by exposure to radiation. In one embodiment, the non-proteinaceous moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation can be of any wavelength, and examples include, but are not limited to, wavelengths that do not harm normal cells but heat the non-proteinaceous moiety to a temperature at which proximal cells of the antibody-non-proteinaceous moiety conjugate die.

[0139] Another modification of the antibodies described herein is pegylation. Antibodies can be pegylated, for example, to increase the biological (e.g., serum) half-life of the antibody. To pegylate an antibody, the antibody, or a fragment thereof, is typically reacted with a PEG, such as a reactive ester or aldehyde derivative of PEG, under conditions such that one or more PEG groups bind to the antibody or antibody fragment. Preferably, the pegylation is carried out via an acylation or alkylation reaction using a reactive PEG molecule (or similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any form of PEG that has been used to derivatize other proteins, such as mono(CI-CIO)alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In some embodiments, the antibody to be pegylated is a non-glycosylated antibody. Methods for pegylating proteins are known in the art and can be applied to the antibodies described herein. See, for example, EP 0 154 316 by Nishimura et al. and EP0401384 by Ishikawa et al.

[0140] The invention also encompasses the human monoclonal antibodies described herein conjugated to a therapeutic agent, a polymer, a detectable label, or an enzyme. In one embodiment, the therapeutic agent is a cytotoxic drug. In one embodiment, the polymer is PEG.

[0141] h. Nucleic Acids, Expression Cassettes, and Vectors The present invention provides isolated nucleic acid segments encoding the polypeptides, peptide fragments, and binding proteins of the present invention. The nucleic acid segments of the present invention also include segments that encode the same amino acids due to the degeneracy of the genetic code. For example, the amino acid threonine is encoded by ACU, ACC, ACA, and ACG and is thus degenerate. The present invention is intended to include all variations of polynucleotide segments that encode the same amino acid. Such mutations are known in the art (Watson et al., Molecular Biology of the Gene, Benjamin Cummings 1987). Mutations also include changes in nucleic acid segments to encode conservative amino acid changes, for example, substitution of leucine for isoleucine. Such mutations are also known in the art. Thus, the genes and nucleotide sequences of the present invention include both naturally occurring sequences and variants.

[0142] The nucleic acid segments of the present invention can be contained within vectors. Vectors can include, but are not limited to, double-stranded or single-stranded linear or circular plasmids, phagemids, F factors, viruses, cosmids, or phages, which may or may not be self-transmissible or mobilizable. The vectors can also transform prokaryotic or eukaryotic hosts by integration into the cell genome or by existing episomally (e.g., autonomously replicating plasmids having an origin of replication).

[0143] Preferably, the nucleic acid segment within the vector is under the control of and operably linked to an appropriate promoter or other regulatory element for transcription in vitro or in a host cell such as a eukaryotic cell or a microorganism, e.g., a bacterium. The vector can be a shuttle vector that functions in multiple hosts. The vector can also be a cloning vector typically containing one or a few restriction endonuclease recognition sites into which a foreign DNA sequence can be inserted in a determinable manner. Such insertions can occur without loss of the essential biological function of the cloning vector. The cloning vector can also contain a marker gene suitable for use in the identification and selection of cells transformed with the cloning vector. Examples of marker genes are tetracycline resistance or ampicillin resistance. Many cloning vectors are commercially available (Stratagene, New England Biolabs, Clonetech).

[0144] The nucleic acid segment of the present invention can also be inserted into an expression vector. Usually, an expression vector contains a prokaryotic DNA element encoding a bacterial origin of replication and an antibiotic resistance gene to provide for amplification and selection of the expression vector in a bacterial host; a regulatory element controlling the initiation of transcription such as a promoter; and a DNA element controlling the processing of the transcript, e.g., an intron, or a transcription termination / polyadenylation sequence.

[0145] Methods for introducing nucleic acid segments into vectors are available in the art (Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (2001)). Briefly, the vector into which the nucleic acid segment is inserted is treated with one or more restriction enzymes (restriction endonucleases) to produce a linearized vector having blunt ends, "sticky ends" with 5' or 3' overhangs, or a combination of any of the above. The vector may also be treated with a restriction enzyme and then with other modifying enzymes such as polymerase, exonuclease, phosphatase or kinase to produce a linearized vector having properties useful for ligation of the nucleic acid segment into the vector. The nucleic acid segment to be inserted into the vector is treated with one or more restriction enzymes to produce a linearized segment having blunt ends, "sticky ends" with 5' or 3' overhangs, or a combination of any of the above. The nucleic acid segment may also be treated with a restriction enzyme and subsequently with other DNA modifying enzymes. Such DNA modifying enzymes include, but are not limited to, polymerase, exonuclease, phosphatase, or kinase for producing a nucleic acid segment having properties useful for ligation of the nucleic acid segment into the vector.

[0146] Next, the processed vector and nucleic acid segment are ligated together according to methods available in the art to form a construct comprising the nucleic acid segment (Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (2001)). Briefly, the processed nucleic acid fragment, and the processed vector are combined in the presence of an appropriate buffer and ligase. The mixture is then incubated under appropriate conditions for the ligase to ligate the nucleic acid fragment to the vector.

[0147] The present invention also provides an expression cassette comprising a nucleic acid sequence capable of directing the expression of a specific nucleic acid segment of the present invention, either in vitro or in a host cell. Further, the nucleic acid segment of the present invention can be inserted into the expression cassette such that an antisense message is generated. The expression cassette is a separable unit such that the expression cassette is in linear form and can be functional for in vitro transcription and translation assays. Materials and procedures for performing these assays are commercially available from Promega (Madison, Wis.). For example, an in vitro transcript can be generated by placing the nucleic acid sequence under the control of a T7 promoter and then using T7 RNA polymerase to generate an in vitro transcript. This transcript can then be translated in vitro by using rabbit reticulocyte lysate. Alternatively, the expression cassette can be incorporated into a vector that also allows for replication and amplification of the expression cassette in a host cell, or in vitro transcription and translation of the nucleic acid segment.

[0148] Such an expression cassette may contain one or more restriction sites that allow the placement of nucleic acid segments under the control of regulatory sequences. The expression cassette may also contain a termination signal operably linked to the nucleic acid segment and regulatory sequences necessary for proper translation of the nucleic acid segment. The expression cassette containing the nucleic acid segment may be chimeric, which means that at least one of its components is heterologous with respect to at least one of the other components. The expression cassette may also be naturally occurring but obtained in a recombinant form useful for heterologous expression. Expression of the nucleic acid segment in the expression cassette may be under the control of a constitutive promoter or an inducible promoter that initiates transcription only when the host cell is exposed to a specific external stimulus. The expression cassette may contain, in the 5' to 3' direction of transcription, a transcription and translation initiation region, a nucleic acid segment, and a transcription and translation termination region that functions in vivo and / or in vitro. The termination region may be specific to the transcription initiation region, may be specific to the nucleic acid segment, or may be derived from another source.

[0149] The regulatory sequences can be polynucleotide sequences located upstream (5' non-coding sequence), within, or downstream (3' non-coding sequence) of the coding sequence, which affect the transcription, RNA processing or stability, or translation of the associated coding sequence. Examples of regulatory sequences include, but are not limited to, enhancers, promoters, repressor binding sites, translation leader sequences, introns, and polyadenylation signal sequences. They can include sequences that can be natural and synthetic sequences, as well as combinations of synthetic and natural sequences. The regulatory sequences are not limited to promoters, but some useful regulatory sequences include constitutive promoters, inducible promoters, regulated promoters, tissue-specific promoters, viral promoters, and synthetic promoters.

[0150] A promoter is a nucleotide sequence that controls the expression of a coding sequence by providing recognition for RNA polymerase and other factors necessary for proper transcription. Promoters include minimal promoters composed only of all the basic elements necessary for transcription initiation, such as the TATA box and / or the initiator, which is a short DNA sequence composed of the TATA box, and other sequences useful for specifying the site of transcription initiation, to which regulatory elements are added for the control of expression. A promoter can be derived entirely from a native gene, or can be composed of different elements derived from different promoters found in nature, or can be composed of synthetic DNA segments. A promoter can include DNA sequences involved in the binding of protein factors that control the effectiveness of transcription initiation in response to physiological or developmental conditions.

[0151] The present invention also provides constructs comprising vectors and expression cassettes. The vector can be selected from any of the above-mentioned vectors, but is not limited thereto. An expression cassette can be inserted into this vector, which is known in the art, via the above-mentioned methods (Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (2001)). In one embodiment, the regulatory sequences of the expression cassette can be derived from a source other than the vector into which the expression cassette is inserted. In other embodiments, a construct comprising a vector and an expression cassette is formed by inserting the nucleic acid segment of the present invention into a vector that itself contains regulatory sequences. Thus, an expression cassette is formed by inserting a nucleic acid segment into a vector. Vectors containing regulatory sequences are commercially available, and methods for using them are known in the art (Clonetech, Promega, Stratagene).

[0152] In other aspects, the disclosure also provides (i) a nucleic acid molecule encoding a polypeptide chain of the antibody or antigen-binding fragment thereof; (ii) a vector comprising the nucleic acid molecule as described; and (iii) a cultured host cell comprising the vector as described. Also provided is a method for producing a polypeptide, comprising (a) obtaining a cultured host cell as described; (b) culturing the cultured host cell in a medium under conditions that allow expression of the polypeptide encoded by the vector and assembly of the antibody or fragment thereof; and (c) purifying the antibody or fragment from the cultured cells or the cell culture medium.

[0153] i. Manufacturing method Antibodies can be produced, for example, using recombinant methods and compositions as described in U.S. Patent No. 4,816,567. In one embodiment, an isolated nucleic acid encoding an antibody described herein is provided. Such a nucleic acid can encode an amino acid sequence comprising the VL of the antibody and / or an amino acid sequence comprising the VH (e.g., the light chain and / or heavy chain of the antibody). In further embodiments, one or more vectors (e.g., expression vectors) comprising such nucleic acids are provided. In further embodiments, host cells comprising such nucleic acids are provided. In such an embodiment, the host cell comprises (e.g., is transformed with): (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antibody, or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and a second vector comprising a nucleic acid encoding an amino acid sequence comprising the VH of the antibody. In one embodiment, the host cell is a eukaryote, such as Chinese hamster ovary (CHO) cells or lymphoid cells (e.g., Y0 cells, NS0 cells, Sp20 cells). In one embodiment, a method for making an antibody is provided, the method comprising culturing a host cell (or host cell culture medium) comprising a nucleic acid encoding the antibody under conditions suitable for expression of the antibody, as described above, and optionally recovering the antibody from the host cell.

[0154] For the recombinant production of antibodies, for example as described above, nucleic acids encoding the antibodies are isolated and inserted into one or more vectors for further cloning and / or expression in host cells. Such nucleic acids can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that can specifically bind to the genes encoding the heavy and light chains of the antibody).

[0155] Host cells suitable for the cloning or expression of vectors encoding antibodies include the prokaryotic or eukaryotic cells described herein. For example, antibodies can be produced in bacteria, particularly when glycosylation and Fc effector functions are not required. For the expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523 (see also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J., 2003), pp. 245-254, which describes the expression of antibody fragments in E. coli). After expression, the antibodies can be isolated from the bacterial cell paste in the soluble fraction and further purified.

[0156] In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeast are suitable cloning or expression hosts for vectors encoding antibodies, such as fungal and yeast strains in which the glycosylation pathway is "humanized" and antibodies with a partial or fully human glycosylation pattern are produced. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006).

[0157] Host cells suitable for the expression of glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. In particular, a number of baculovirus strains that can be used in combination with insect cells have been identified for the transfection of Spodoptera frugiperda cells.

[0158] Plant cell cultures can also be utilized as hosts. See, for example, U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (which describe the PLANTIBODIES technology for producing antibodies in transgenic plants). 。

[0159] Vertebrate cells can also be used as hosts. For example, mammalian cell lines adapted to grow in suspension can be useful. Other examples of useful mammalian host cell lines include the monkey kidney CV1 cell line transformed by SV40 (COS-7); human fetal kidney cell lines (e.g., 293 or 293 cells described in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse Sertoli cells (e.g., TM4 cells as described in Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical cancer cells (HELA); dog kidney cells (MDCK); buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells as described, for example, in Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC5 cells; and FS4 cells. Other useful mammalian host cell lines include CHO cells such as DHFR-CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NS0, and Sp2 / 0. For a review of specific mammalian host cell lines suitable for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J.), pp. 255-268 (2003).

[0160] B. Compositions and Formulations In other aspects, the disclosure also provides a pharmaceutical composition comprising (i) at least one anti-HIV antibody or antigen-binding portion thereof, nucleic acid, or vector as described above; and (ii) a pharmaceutically acceptable carrier.

[0161] In some embodiments, the pharmaceutical composition further comprises a second therapeutic agent. In some embodiments, the second therapeutic agent comprises an antiviral agent or one or more additional antibodies. In some embodiments, the one or more additional antibodies comprise a second anti-HIV antibody (e.g., an isolated anti-HIV bNAb as disclosed) or an antigen-binding portion thereof, or a third antibody that binds to a third antigen.In some embodiments, the third antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-l), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), CD160 (NK1), immune checkpoint proteins (e.g., PD-1, PD-L1, CTLA-4), and the second epitope of gp120.

[0162] In some embodiments, the antiviral agent is selected from the group consisting of non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry inhibitors or fusion inhibitors, and integrase inhibitors.

[0163] The pharmaceutical composition of the present invention can also be administered as a vaccine (e.g., as an AAV-based vaccine), or in combination therapy with, for example, other immunostimulants, antiviral agents, or vaccines. In some embodiments, the composition comprises the antibody of the present invention at a concentration of at least 1 mg / ml, 5 mg / ml, 10 mg / ml, 50 mg / ml, 100 mg / ml, 150 mg / ml, 200 mg / ml, 1 - 300 mg / ml, or 100 - 300 mg / ml.

[0164] In some embodiments, the second therapeutic agent comprises an anti-inflammatory drug or an antiviral compound. In some embodiments, the antiviral compound comprises a nucleoside analog, a peptoid, an oligopeptide, a polypeptide, a protease inhibitor, a 3C-like protease inhibitor, a papain-like protease inhibitor, or an inhibitor of RNA-dependent RNA polymerase. In some embodiments, examples of the antiviral compound may include acyclovir, ganciclovir, vidarabine, foscarnet, cidofovir, amantadine, ribavirin, trifluorothymidine, zidovudine, didanosine, zalcitabine, or interferon. In some embodiments, the interferon is interferon-α or interferon-β.

[0165] The use of the pharmaceutical composition in the preparation of a medicament for the diagnosis, prevention, treatment, or combination thereof of conditions resulting from HIV-1 infection is also within the scope of the present disclosure.

[0166] The pharmaceutical composition may contain any number of excipients. Excipients that may be used include carriers, surfactants, thickeners or emulsifiers, solid binders, dispersion or suspension aids, solubilizers, colorants, flavors, coatings, disintegrants, lubricants, sweeteners, preservatives, isotonic agents, and combinations thereof. The selection and use of appropriate additives are taught in Gennaro, ed., Remington: The Science and Practice of Pharmacy, 20th Ed. (Lippincott Williams & Wilkins 2003), the disclosure of which is incorporated herein by reference.

[0167] Preferably, the pharmaceutical composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound can be coated with a material to protect it from the action of acids and other natural conditions that can inactivate it. As used herein, the term "parenteral administration" generally means a mode of administration other than enteral and topical administration by injection and includes, but is not limited to, intravenous, intramuscular, arterial, intracavitary, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injections and infusions. Alternatively, the antibodies of the invention described herein can be administered via local, epidermal or mucosal routes of administration, e.g., parenteral routes such as intranasal, oral, vaginal, rectal, sublingual or topical.

[0168] The pharmaceutical compositions of the invention can be prepared in many forms, including tablets, hard or soft gelatin capsules, aqueous solutions, suspensions, liposomes, and other sustained release formulations such as shaped polymeric gels. Oral dosage forms can be formulated such that the antibody is released into the intestine after passing through the stomach. Such formulations are described in U.S. Patent No. 6,306,434 and the references contained therein.

[0169] Oral liquid pharmaceutical compositions can be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or can be presented as dry products for constitution with water or other suitable vehicles before use. Such liquid pharmaceutical compositions can contain conventional additives such as suspending agents, emulsifying agents, non-aqueous vehicles (which can include edible oils), or preservatives.

[0170] Antibodies can be formulated for parenteral administration (e.g., by injection, e.g., by bolus injection or continuous infusion) and can be presented in unit dosage form in ampoules, prefilled syringes, small volume infusion containers, or multi-dose containers with added preservatives. The pharmaceutical composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle and can contain formulation agents such as suspending agents, stabilizers and / or dispersing agents. Pharmaceutical compositions suitable for rectal administration can be prepared as unit dose suppositories. Suitable carriers include physiological saline and other materials commonly used in the art.

[0171] For administration by inhalation, the antibody can be conveniently delivered from a nebulizer, atomizer, or pressurized pack, or other convenient means for delivering an aerosol spray. The pressurized pack can contain a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the unit of dosage can be determined by providing a valve to supply a metered amount.

[0172] Alternatively, for administration by inhalation or insufflation, the antibody can be in the form of a dry powder composition, for example, a powder mixture with a suitable powder base such as a modulator and lactose or starch. The powder composition can be presented in unit dosage form in, for example, capsules or cartridges, or in gelatin or blister packs from which the powder can be administered, for example, with the aid of an inhaler or insufflator. For nasal administration, the antibody can be administered via a liquid spray such as a plastic bottle atomizer.

[0173] The pharmaceutical composition of the present invention may also contain other components such as flavoring agents, coloring agents, antibacterial agents, or preservatives. It will be understood that the amount of antibody required for treatment will vary not only with the particular carrier selected, but also with the route of administration, the nature of the condition being treated, and the age and condition of the patient. Ultimately, the attending healthcare provider may determine the appropriate dosage. Further, the pharmaceutical composition may be formulated as a single unit dosage form.

[0174] The pharmaceutical composition of the present invention can be in the form of a sterile aqueous solution or dispersion. It can also be formulated in microemulsions, liposomes, or other ordered structures suitable for high drug concentrations.

[0175] The antibodies of the present invention described herein can be administered as sustained release formulations, in which case less frequent dosing is required. The dosage and dosing frequency vary depending on the half-life of the antibody in the patient. Generally, human antibodies exhibit the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. The dosage and dosing frequency can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic use, relatively low doses are administered at relatively infrequent intervals over a long period of time. Some patients may receive treatment throughout their lives. In therapeutic use, relatively high doses may be required at relatively short intervals until the progression of the disease is reduced or terminated, preferably until the patient shows partial or complete improvement of the symptoms of the disease. Thereafter, the patient may be administered a prophylactic regimen.

[0176] The amount of active ingredient that can be combined with the carrier material to produce a single dosage form varies depending on the subject being treated and the particular mode of administration, and is usually that amount of the composition that produces a therapeutic effect. Generally, out of 100%, this amount ranges from about 0.01% to about 99% of the active ingredient, preferably from about 0.1% to about 70% of the active ingredient, and most preferably from about 1% to about 30% of the active ingredient, in combination with a pharmaceutically acceptable carrier.

[0177] The pharmaceutical composition can be a controlled release formulation, such as an implant, a transdermal patch, and a microencapsulation delivery system. Biodegradable and biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. See, for example, Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0178] The therapeutic composition can be administered via medical devices such as (1) needleless subcutaneous injection devices (e.g., US 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; and 4,596,556); (2) microinfusion pumps (US 4,487,603); (3) transdermal devices (US 4,486,194); (4) injection devices (US 4,447,233 and 4,447,224); and (5) osmotic devices (US 4,439,196 and 4,475,196), etc., the disclosures of which are hereby incorporated by reference herein.

[0179] In some embodiments, the human monoclonal antibodies of the invention described herein can be formulated to ensure proper distribution in vivo. For example, to ensure that the therapeutic compounds of the invention cross the blood-brain barrier, they can be formulated into liposomes, which may further include targeting moieties for enhancing selective delivery to specific cells or organs. See, for example, US 4,522,811; 5,374,548; 5,416,016; and 5,399,331; V.V. Ranade (1989) Clin. Pharmacol. 29:685; Umezawa et al., (1988) Biochem. Biophys. Res. Commun. 153:1038; Bloeman et al. (1995) FEBS Lett. 357:140; M. Owais et al. (1995) Antimicrob. Agents Chemother. 39:180; Briscoe et al. (1995) Am. Physiol. 1233:134; Schreier et al. (1994). Biol. Chem. 269:9090; Keinanen and Laukkanen (1994) FEBS Lett. 346:123; and Killion and Fidler (1994) Immunomethods 4:273.

[0180] In some embodiments, after the first dose, subsequent doses of the antibody or antigen-binding fragment thereof, which may be approximately the same as or less than the first dose, can be administered at regular intervals of at least 1 to 3 days; at least 1 week, at least 2 weeks; at least 3 weeks; at least 4 weeks; at least 5 weeks; at least 6 weeks; at least 7 weeks; at least 8 weeks; at least 9 weeks; at least 10 weeks; at least 12 weeks; or at least 14 weeks.

[0181] A variety of delivery systems are known and can be used to administer the pharmaceutical compositions of the present invention, for example, encapsulation into liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see, e.g., Wu et al. (1987) J. Biol. Chem. 262:4429-4432). Methods of introduction include, but are not limited to, intradermal, transdermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, for example, by infusion or bolus injection, absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be administered together with other biologically active agents. Administration can be systemic or local. The pharmaceutical compositions can also be carried in vesicles, particularly liposomes (see, e.g., Langer (1990) Science 249: 1527-1533).

[0182] The use of nanoparticles for delivering the antibodies of the present invention is also contemplated herein. Antibody-conjugated nanoparticles can be used for both therapeutic and diagnostic applications. Antibody-conjugated nanoparticles and methods of preparation and use are described in detail by Arruebo, M., et al. 2009 (“Antibody-conjugated nanoparticles for biomedical applications” in J. Nanomat. Volume 2009, Article ID 439389), which is incorporated herein by reference. Nanoparticles can be developed and conjugated to antibodies included in pharmaceutical compositions to target cells. Nanoparticles for drug delivery are also described, for example, in US 8257740, or US 8246995, each of which is incorporated herein by reference in its entirety.

[0183] Under specific conditions, the pharmaceutical composition can be delivered by a controlled release system. In one embodiment, a pump can be used. In other embodiments, polymeric materials can be used. In still other embodiments, the controlled release system can be placed near the target of the composition, and thus only a small portion of the systemic dose is required.

[0184] The injectable preparation can include dosage forms for intravenous, subcutaneous, intradermal, intracranial, intraperitoneal, and intramuscular injections, infusion, etc. These injectable preparations can be prepared by known methods. For example, the injectable preparation can be prepared by dissolving, suspending, or emulsifying the above-mentioned antibody or its salt in a sterile aqueous medium or an oily medium conventionally used for injection. Examples of the aqueous medium for injection include, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, which can be used in combination with appropriate solubilizing agents such as alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)]. As the oily medium, for example, sesame oil, soybean oil, etc. are used and can be used in combination with solubilizing agents such as benzyl benzoate, benzyl alcohol. The injection thus prepared is preferably filled into appropriate ampoules.

[0185] The pharmaceutical composition of the present invention can be delivered subcutaneously or intravenously using standard needles and syringes. Further, with respect to subcutaneous delivery, pen delivery devices can readily have use in delivering the pharmaceutical composition of the present invention. Such pen delivery devices can be reusable or disposable. Reusable pen delivery devices typically utilize replaceable cartridges containing the pharmaceutical composition. Once all of the pharmaceutical composition within the cartridge has been administered and the cartridge is empty, the empty cartridge can be readily discarded and replaced with a new cartridge containing the pharmaceutical composition. Thereafter, the pen delivery device can be reused. Disposable pen delivery devices do not have replaceable cartridges. Rather, the disposable pen delivery device is pre-filled with the pharmaceutical composition held in a reservoir within the device. When the pharmaceutical composition runs out from the reservoir, the entire device is discarded.

[0186] A number of reusable pens and auto-injector delivery devices have use in the subcutaneous delivery of the pharmaceutical compositions of the present invention. By way of example only, and not by way of limitation, these include AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pens (Disetronic Medical Systems, Burghdorf, Switzerland), HUMALOG MIX 75 / 25™ pen, HUMALOG™ pen, HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II, and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (Sanofi-Aventis, Frankfurt, Germany). Examples of disposable pen delivery devices having use in the subcutaneous delivery of the pharmaceutical compositions of the present invention include, by way of example only, and not by way of limitation, SOLOSTAR™ pen (Sanofi-Aventis), FLEXPEN™ (Novo Nordisk), and KWIKPEN™ (Eli Lilly), SURECLICK™ auto-injector (Amgen, Thousand Oaks, CA), PENLET™ (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, L.P.), and HUMIRA™ pen (Abbott Labs, Abbott Park, IL).

[0187] Advantageously, the pharmaceutical composition for oral or parenteral use described above is prepared in a unit dosage form suitable for accommodating the dosage of the active ingredient. Such dosage forms in unit dosage include, for example, tablets, pills, capsules, injections (ampoules), suppositories and the like. The amount of the antibody contained is usually about 5 mg to about 500 mg per dosage form in unit dosage; particularly in the form of injection, the antibody preferably contains about 5 to about 300 mg, and in other dosage forms, it preferably contains about 10 mg to about 300 mg.

[0188] C. Methods and Uses a. Treatment Methods According to other embodiments, the present invention provides a method for treating a mammal infected with a virus (e.g., HIV) comprising administering to the mammal a pharmaceutical composition comprising an anti-HIV antibody disclosed herein. According to one embodiment, a method for treating a mammal infected with HIV comprises administering to the mammal a pharmaceutical composition comprising an antibody or a fragment thereof of the present invention. The compositions of the present invention may comprise two or more antibodies (e.g., a plurality of antibodies or a pool of antibodies) having the disclosed characteristics. It may also include other HIV neutralizing antibodies known in the art, such as GS-9722 (elipovimab), PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT-136, PGT-137, PGT-138, PGT-139, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 354BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1, and VRC29.03 (all of which bind to the V3 glycan region), 2F5, 4E10, M66.6, CAP206-CH12, 10E8, 10E8v4, 10E8-5R-100cF, DH511.11P, 7b2, and LN01 (all of which bind to the MPER of gp41); PG9, PG16, CH01-04 (all of which bind to V1V2-glycan), 2G12 (binds to the external domain glycan), VRC01, PG9, and b12, but is not limited thereto.

[0189] In some embodiments, the present disclosure provides a method for preventing or treating HIV infection or an HIV-related disease. The method comprises (a) identifying a patient in need of said prevention or treatment, and (b) administering to said patient a first therapeutic agent comprising a therapeutically effective amount of at least one anti-HIV antibody or an antigen-binding portion thereof as described above.

[0190] In some embodiments, the method further comprises administering a second therapeutic agent. In some embodiments, the second therapeutic agent comprises an antiviral agent or one or more additional antibodies. In some embodiments, the one or more additional antibodies comprise a second anti-HIV antibody (e.g., an isolated anti-HIV bNAb as disclosed) or an antigen-binding portion thereof, or a third antibody that binds to a third antigen.In some embodiments, the third antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-l), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), CD160 (NK1), immune checkpoint proteins (e.g., PD-1, PD-L1, CTLA-4), and the second epitope of gp120.

[0191] In some embodiments, the antiviral agent is selected from the group consisting of non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry inhibitors or fusion inhibitors, and integrase inhibitors.

[0192] In some embodiments, the first therapeutic agent or the second therapeutic agent is administered to the patient intratumorally, intravenously, subcutaneously, intraosseously, orally, transdermally, or sublingually. In some embodiments, the first therapeutic agent is administered to the patient before, after, or simultaneously with the second therapeutic agent. In some embodiments, the antibody or antigen-binding fragment thereof is administered prophylactically or therapeutically.

[0193] Passive immunization has proven to be an effective and safe strategy for the prevention and treatment of viral diseases. (See, e.g., Keller et al., Clin. Microbiol. Rev. 13:602-14 (2000); Casadevall, Nat. Biotechnol. 20:114 (2002); Shibata et al., Nat. Med. 5:204-10 (1999); and Igarashi et al., Nat. Med. 5:211-16 (1999), each incorporated herein by reference). Passive immunization using human monoclonal antibodies provides an immediate treatment strategy for the emergency prevention and treatment of HIV. Subjects at risk of HIV-related diseases or disorders include patients who have been in contact with an infected individual or who have otherwise been exposed to HIV. The prophylactic agent can be administered before the symptoms characteristic of an HIV-related disease or disorder appear, such that the disease or disorder is prevented or its progression is delayed.

[0194] For in vivo treatment of human and non-human patients, the patient is administered or provided with a formulation comprising the HIV antibody of the present invention. When used for in vivo treatment, the antibody of the present invention is administered to the patient in a therapeutically effective amount (i.e., an amount that eliminates or reduces the amount of virus in the patient). The antibody is administered to human patients according to known methods such as intravenous administration, for example, as a bolus or by continuous infusion over a period of time, intramuscularly, intraperitoneally, intraspinally, subcutaneously, intra-articularly, intrasynovially, intrathecally, orally, topically, or by the inhalation route. The antibody can be administered parenterally or intravenously, if possible, to the target cell site. In some embodiments, the antibody is administered by intravenous or subcutaneous administration. The therapeutic composition of the present invention can be administered systemically, parenterally, or topically to a patient or subject. The above parameters for evaluating the success and improvement of the treatment of the disease can be easily measured by routine procedures well known to physicians.

[0195] For parenteral administration, the antibody can be formulated in unit dose injectable form (solution, suspension, emulsion) in combination with a pharmaceutically acceptable parenteral vehicle. Examples of such vehicles include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Examples of non-aqueous vehicles include, but are not limited to, fixed oils and ethyl oleate. Liposomes can be used as carriers. The vehicle can contain small amounts of additives such as substances that enhance isotonicity and chemical stability, such as buffers and preservatives. The antibody can be formulated in such a vehicle at a concentration of about 1 mg / ml to 10 mg / ml.

[0196] The dosage and dosing schedule depend on various factors that can be readily determined by a physician, such as the nature of the infection, e.g., its therapeutic index, the patient, and the patient's medical history. Usually, a therapeutically effective amount of the antibody is administered to the patient. In some embodiments, the amount of the antibody administered ranges from about 0.1 mg / kg to about 50 mg / kg of the patient's body weight. Depending on the type and severity of the infection, about 0.1 mg / kg to about 50 mg / kg body weight of the antibody (e.g., about 0.1 - 15 mg / kg / dose) is a first candidate dosage for administration to the patient, whether by, for example, one or more separate administrations or by continuous infusion. The progress of this treatment is readily monitored by a physician or a person of ordinary skill in the art based on criteria known by conventional methods and assays. The above parameters for assessing the success and improvement of the treatment of the disease are readily measurable by routine procedures well known to physicians.

[0197] Other treatment regimens can be combined with the administration of the HIV antibodies of the present invention. Such combination administrations include co - administrations using separate formulations or a single formulation, and sequential administrations in either order, preferably with a period during which both (or all) of the active agents exert their biological activities simultaneously. Such combination therapies can result in a synergistic therapeutic effect. The above parameters for assessing the success and improvement of the treatment of the disease are readily measurable by routine procedures well known to physicians.

[0198] b. Method of reducing virus replication A method is further provided for reducing an increase in the HIV virus titer, virus replication, virus growth, or the amount of HIV viral protein in a subject. According to another aspect, the method comprises administering to the subject an effective amount of an HIV antibody to reduce an increase in the HIV titer, virus replication, or the amount of HIV protein of one or more HIV strains or isolates in the subject.

[0199] According to another embodiment, the present invention provides a method for reducing viral replication or the spread of HIV infection to additional host cells or tissues, which includes contacting an antigenic epitope, for example, an antigenic epitope on gp120, with an antibody or a portion thereof that binds to the antigenic epitope and mammalian cells.

[0200] c. Combination therapy (1) Combination therapy with two or more anti-HIV antibodies In some embodiments, the present disclosure provides a method for treating or preventing HIV infection in a human subject having or at risk of having HIV infection. The method includes administering to the human subject a therapeutically effective amount of an antibody or antigen-binding fragment, or a pharmaceutical composition thereof, as disclosed herein, in combination with a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or 1 - 3) additional therapeutic agents. In one embodiment, a method for treating HIV infection in a human subject suffering from or at risk of suffering from an infectious disease is provided, the method including administering to the human subject a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or 1 - 3) additional therapeutic agents in combination with a therapeutically effective amount of one or more antibodies, or pharmaceutically acceptable salts thereof, as disclosed herein.

[0201] Antibody combination therapy In some embodiments, the disclosed antibody or antigen-binding fragment thereof is co-administered with a second anti-HIV antibody. In some embodiments, the antibody or antigen-binding fragment thereof is co-administered with a second anti-HIV antibody that binds to an epitope or region of gp120 selected from the group consisting of: (i) the third variable loop (V3) and / or the high-mannose patch containing N332 oligomannose glycans; (ii) the second variable loop (V2) and / or the apex of the Env trimer; (iii) the CD4 binding site (CD4b); (iv) the gp120 / gp41 interface; or (v) the silent face of gp120. The aforementioned epitopes or regions of gp120 bound by broad neutralizing antibodies are described, for example, in McCoy, Retrovirology (2018) 15:70; Sok and Burton, Nat Immunol. 2018 19(11):1179-1188; Possas, et al., Expert Opin Ther Pat. 2018 Jul;28(7):551-560; and Stephenson and Barouch, Curr HIV / AIDS Rep (2016) 13:31-37, which are hereby incorporated by reference in their entirety for all purposes.

[0202] In some embodiments, the combination therapy involves co - administration of an antibody or an antigen - binding fragment thereof with another anti - HIV broadly neutralizing antibody or bNAb (i.e., a neutralizing antibody that neutralizes multiple HIV - 1 virus strains). A variety of bNAbs are known in the art and can be used as combination therapeutics. Exemplary additional bNAbs for use include those that bind to or compete with VH and VL that bind to epitopes or regions of gp120 selected from the group consisting of: (i) the third variable loop (V3) and / or the high - mannose patch containing N332 oligomannose glycan; (ii) the second variable loop (V2) and / or the apex of the Env trimer; (iii) the CD4 - binding site (CD4b); (iv) the gp120 / gp41 interface; or (v) the silent face of gp120.Exemplary bNAbs for use in anti-HIV antibody combination therapies include GS-9722 (elipovimab), PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT-136, PGT-137, PGT-138, PGT-139, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 354BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1 and VRC29.03 (all of which bind to the V3 glycan region), 2F5, 4E10, M66.6, CAP206-CH12, 10E8, 10E8v4, 10E8-5R-100cF, DH511.11P, 7b2, and LN01 (all of which bind to the MPER of gp41); PG9, PG16, CH01-04 (all of which bind to V1V2-glycan), 2G12 (binds to the glycan of the outer domain); b12, F105, VRC01, VRC07, VRC07-523, VRC03, VRC06, VRC06b01 VRC08, VRC0801, NIH45-46, GS-9723, GS-5423, 3BNC117, 3BNC60, VRC-PG04, PGV04; CH103, 44-VRC13.01, 1NC9, 12A12, N6, N6LS (VRC-HIVMAB091-00-AB), N49-P7, NC-Cow1, IOMA, CH235 and CH235.12, N49P6, N49P7, N49P11, N49P9 and N60P25 (all of which bind to the CD4 binding site), including those containing VH and VL that bind or compete with them.

[0203] In some embodiments, the combination therapy includes N332 oligomannose glycans and antibodies selected from the group consisting of GS-9722 (elipovimab), PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT-136, PGT-137, PGT-138, PGT-139, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 354BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1, and VRC29.03, which compete with or contain VH and VL regions from these antibodies, and antibodies that bind to epitopes or regions of gp120 that include the third variable loop (V3) and / or high-mannose patches. Additional broadly neutralizing antibodies that can be used in the second antigen-binding domain of the multispecific antigen-binding molecules described herein, which bind to gp120 of the third variable loop (V3) and / or high-mannose patches containing N332 oligomannose glycans, are described, for example, in WO 2012 / 030904; WO 2014 / 063059; WO 2016 / 149698; WO 2017 / 106346; WO 2018 / 075564, WO 2018 / 125813; WO 2018 / 237148, WO 2019 / 226829, WO 2020 / 023827, WO2020 / 056145, and Kerwin, et al., J Pharm Sci. 2020 Jan;109(1):233-246, which are hereby incorporated by reference in their entirety for all purposes.

[0204] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of gp120 at the CD4 binding site (CD4bs) and competes with or includes the VH and VL regions of antibodies selected from the group consisting of b12, F105, VRC01, VRC07, VRC07-523, VRC03, VRC06, VRC06b01 VRC08, VRC0801, NIH45-46, 3BNC117, GS-9723, GS-5423, 3BNC60, VRC-PG04, PGV04; CH103, 44-VRC13.01, 1NC9, 12A12, N6, N49-P7, NC-Cow1, IOMA, CH235, and CH235.12, N49P6, N49P7, N49P11, N49P9, and N60P25. Additional broadly neutralizing antibodies that bind to gp120 at the CD4 binding site (CD4b) and can be used in the second antigen-binding domain of the multispecific antigen-binding molecules described herein are described, for example, in WO 2011 / 038290; WO 2012 / 158948; WO 2013 / 016468; WO 2013 / 192589; WO 2013 / 086533; WO 2015 / 128846; WO 2016 / 149698; WO 2016 / 149695; WO 2018 / 075564; WO 2018 / 125813; WO 2018 / 237357, and U.S. Patent Nos. 9,493,549, and 9,879,068. Additional antibodies that bind to an epitope or region of gp120 at CD4b and can be administered in combination with or simultaneously with the present antibody are described, for example, in Schommers, et al., Cell (2020) 180: 471-489; Freund, et al., Sci Transl Med (2017) 9: eaal2144; Diskin, et al., J Exp Med (2013) 210: 1235-49; and Scheid, et al., Science (2011) 333: 1633-1637. The foregoing publications are hereby incorporated by reference in their entirety for all purposes.

[0205] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of gp120 of the second variable loop (V2) and / or the Env trimer apex and competes with or comprises the CDR and / or VH and VL regions of antibodies selected from the group consisting of PG9, PG16, PGC14, PGG14, PGT-142, PGT-143, PGT-144, PGT-145, CH01, CH59, PGDM1400, CAP256, CAP256-VRC26.08, CAP256-VRC26.09, CAP256-VRC26.25, PCT64-24E, and VRC38.01.

[0206] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of gp120 of the gp120 / gp41 interface and competes with or comprises the CDR and / or VH and VL regions of antibodies selected from the group consisting of PGT-151, CAP248-2B, 35O22, 8ANC195, ACS202, VRC34, and VRC34.01.

[0207] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of the gp120 silent face and competes with or comprises the second VH and VL regions of antibody VRC-PG05.

[0208] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of gp41 in the membrane-proximal external region (MPER) and competes with or comprises the second VH and VL regions of antibodies selected from the group consisting of 10E8, 10E8v4, 10E8-5R-100cF, 4E10, DH511.11P, 2F5, 7b2, and LN01. In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of KLIC, an invariant site of the transmembrane protein gp41 (“KLIC”), and competes with or comprises the second VH and VL regions of the Clone3 human monoclonal antibody (Cl3hmAb) (Protheragen). See, e.g., Vanini, et al., AIDS. (1993) 7(2):167-74.

[0209] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of a gp41 fusion peptide and competes with or comprises the second VH and VL regions of antibodies selected from the group consisting of VRC34 and ACS202.

[0210] In some embodiments, the combination therapy includes a multispecific, e.g., bispecific or trispecific, antibody that binds to an HIV antigen. Examples of HIV bispecific and trispecific antibodies include MGD014, B12BiTe, BiIA-SG, TMB bispecific, SAR-441236, VRC-01 / PGDM-1400 / 10E8v4, 10E8.4 / iMab, and 10E8v4 / PGT121-VRC01.

[0211] Prior to administration, bNAbs can be improved to have enhanced drug-like properties, reduced immunogenicity, enhanced ADCC, and appropriate pharmacokinetic properties. Such antibodies have been shown to bind to HIV envelope glycoproteins expressed on the surface of virions or infected cells, mediating both direct neutralization of the virus and killing of these cells by potent NK, monocytes, and PBMCs. Due to this property, the antibodies can treat HIV infection by neutralizing the virus, kill and eliminate potentially HIV-infected cells in infected individuals, and lead to sterilizing treatment of HIV.

[0212] In various embodiments, all antibodies administered in combination with anti-HIV antibody therapy can have Fc and / or post-translational modifications that extend serum half-life and / or enhance effector activity, as described above.

[0213] In various embodiments, the antibody or antigen-binding fragment, and optionally the combined bNAb, can be delivered in vivo and can be expressed in vivo, for example, from administered mRNA or engineered B cells. Examples of bNAbs delivered in vivo include AAV8-VRC07; mRNA encoding the anti-HIV antibody VRC01; and engineered B cells encoding 3BNC117 (Hartweger et al., J. Exp. Med. 2019, 1301).

[0214] (2) Combination therapy with other anti-HIV therapeutic agents In some embodiments, methods are provided for treating or preventing HIV infection in a human who has or is at risk of having an infectious disease, comprising administering to the human a therapeutically effective amount of an antibody or antigen-binding fragment in combination with a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents, as disclosed herein. In one embodiment, a method is provided for treating HIV infection in a human who has or is at risk of having an infectious disease, comprising administering to the human a therapeutically effective amount of an antibody or antigen-binding fragment in combination with a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents, as disclosed herein.

[0215] In one embodiment, a pharmaceutical composition is provided comprising an antibody or antigen-binding fragment as disclosed herein, in combination with one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents and a pharmaceutically acceptable carrier, diluent, or excipient.

[0216] In some embodiments, methods are provided for treating HIV infection, comprising administering to a patient in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof in combination with a therapeutically effective amount of one or more additional therapeutic agents suitable for the treatment of HIV infection, as described herein.

[0217] In some embodiments, the antibody or antigen-binding fragment thereof is combined with one, two, three, four, or more additional therapeutic agents. In some embodiments, the antibody or antigen-binding fragment thereof is combined with two additional therapeutic agents. In other embodiments, the antibody or antigen-binding fragment thereof is combined with three additional therapeutic agents. In further embodiments, the antibody or antigen-binding fragment thereof is combined with four additional therapeutic agents. The one, two, three, four, or more additional therapeutic agents can be different therapeutic agents selected from the same class of therapeutic agents (e.g., one or more anti-HIV broadly neutralizing antibodies), and / or they can be selected from different classes of therapeutic agents.

[0218] Administration of combination HIV therapy In some embodiments, as described herein, the antibody or antigen-binding fragment thereof is co-administered with one or more additional therapeutic agents. Co-administration of the antibody or antigen-binding fragment disclosed herein with one or more additional therapeutic agents generally refers to co- or sequential administration of the antibody or antigen-binding fragment disclosed herein and one or more additional therapeutic agents such that both a therapeutically effective amount of the antibody or antigen-binding fragment disclosed herein and one or more additional therapeutic agents are present in the body of the patient. When administered sequentially, the combination can be administered in more than one dose.

[0219] As for co - administration, as described herein, it includes co - administration of the antibody or its antigen - binding fragment before or after administration of a unit dose of one or more additional therapeutic agents, and administration of a unit dose. For example, the antibody or its antigen - binding fragment described herein can be administered within seconds, minutes, hours or days of administration of one or more additional therapeutic agents. In some embodiments, the unit dose of the antibody or antigen - binding fragment disclosed herein is administered first, and then, within seconds, minutes, hours or days, the unit dose of one or more additional therapeutic agents is administered. Alternatively, the unit dose of one or more additional therapeutic agents is administered first, followed by the unit dose of the antibody or antigen - binding fragment disclosed herein within seconds, minutes, hours or days. In other embodiments, the unit dose of the antibody or antigen - binding fragment disclosed herein is administered first, and then, several hours (e.g., 1 - 12 hours, 1 - 24 hours, 1 - 36 hours, 1 - 48 hours, 1 - 60 hours, 1 - 72 hours) later, the unit dose of one or more additional therapeutic agents is administered. In still other embodiments, the unit dose of one or more additional therapeutic agents is administered first, and then, several hours (e.g., 1 - 12 hours, 1 - 24 hours, 1 - 36 hours, 1 - 48 hours, 1 - 60 hours, 1 - 72 hours) later, the unit dose of the antibody or antigen - binding fragment disclosed herein is administered.

[0220] In some embodiments, the antibody or antigen - binding fragment disclosed herein is combined with one or more additional therapeutic agents in a single dosage form, such as a solid, liquid or suspension dosage form, for oral, intravenous, intramuscular or subcutaneous administration, for co - administration to a patient.

[0221] In some embodiments, the antibody or antigen-binding fragment is formulated as a liquid solution or suspension and may optionally contain one or more other compounds useful for the treatment of HIV. In some embodiments, the liquid solution or suspension may contain other active ingredients for the treatment of HIV such as HIV protease inhibitors, HIV non-nucleoside or non-nucleotide inhibitors of reverse transcriptase, HIV nucleoside or nucleotide inhibitors of reverse transcriptase, HIV integrase inhibitors, HIV non-catalytic site (or allosteric) integrase inhibitors, pharmacokinetic enhancers, and combinations thereof.

[0222] In some embodiments, such liquid solutions or suspensions are suitable for dosing or administration intervals of once a day, once a week (i.e., QW), once every two weeks (i.e., every other week, or once every two weeks, or Q2W), once a month (i.e., QM), or once every two months (i.e., every other month, or once every two months, or Q2M). In some embodiments, the antibody or antigen-binding fragment is administered once a day, once a week (i.e., QW), once every two weeks (i.e., every other week, or once every two weeks, or Q2W), once a month (i.e., QM), once every two months, once every three months (i.e., Q3M), once every four months (i.e., Q4M).

[0223] HIV combination therapy In the above-described embodiment, the additional therapeutic agent can be an anti-HIV agent. HIV protease inhibitor, non-nucleoside or non-nucleotide inhibitor of reverse transcriptase of HIV, nucleoside or nucleotide inhibitor of reverse transcriptase of HIV, HIV integrase inhibitor, HIV non-catalytic site (or allosteric) integrase inhibitor, HIV entry inhibitor, HIV maturation inhibitor, HIV capsid inhibitor, HIV Tat or Rev inhibitor, immunomodulatory agent (e.g., immunostimulant), immunotherapeutic agent, immunomodulatory agent, immunotherapeutic agent, antibody-drug conjugate, gene modification factor, gene editor (CRISPR / Cas9, zinc finger nuclease, homing nuclease, synthetic nuclease, TALEN, etc.), cell therapy (chimeric antigen receptor T cells, CAR-T, and engineered T cell receptor, TCR-T, autologous T cell therapy, etc.), latency reversing agent, compound targeting HIV capsid, immune-based therapy, phosphatidylinositol 3-kinase (PI3K) inhibitor, HIV antibody, bispecific antibody and "antibody-like" therapeutic protein, HIV p17 matrix protein inhibitor, IL-13 antagonist, peptidylprolyl cis-trans isomerase A modulator, protein disulfide isomerase inhibitor, complement C5a receptor antagonist, DNA methyltransferase inhibitor, fatty acid synthase inhibitor, HIV vif gene modulator, Vif dimerization antagonist, HIV-1 viral infectivity factor inhibitor, TAT protein inhibitor, HIV-1 Nef modulator (e.g., Nef inhibitor), Hck tyrosine kinase modulator, mixed lineage kinase-3 (MLK-3) inhibitor, HIV-1 splicing inhibitor, Rev protein inhibitor, integrin antagonist, nucleoprotein inhibitor, splicing factor modulator, COMM domain-containing protein 1 modulator, HIV ribonuclease H inhibitor, retrocyclin modulator, CDK-4 inhibitor, CDK-6 inhibitor, CDK-9 inhibitor, dendritic cell ICAM-3-grabbing non-integrin 1 inhibitor, HIV GAG protein inhibitor, HIVPOL protein inhibitor, complement factor H modulator, ubiquitin ligase inhibitor, deoxycytidine kinase inhibitor, cyclin-dependent kinase inhibitor, proprotein convertase PC9 stimulator, ATP-dependent RNA helicase DDX3X inhibitor, reverse transcriptase priming complex inhibitor, G6PD and NADH-oxidase inhibitor, mTOR complex 1 inhibitor, mTOR complex 2 inhibitor, P-glycoprotein modulator, TAT protein inhibitor, prolidase inhibitor, phospholipase A2 inhibitor, pharmacokinetic enhancer, HIV gene therapy, TNFα ligand inhibitor, IFN antagonist, HIV vaccine, and combinations thereof.

[0224] In some embodiments, the additional therapeutic agent is selected from the group consisting of HIV combination drugs, other drugs for treating HIV, HIV protease inhibitors, HIV reverse transcriptase inhibitors, HIV integrase inhibitors, HIV non-catalytic site (or allosteric) integrase inhibitors, HIV entry (fusion) inhibitors, HIV maturation inhibitors, latency reversing agents, HIV capsid inhibitors, HIV Tat or Rev inhibitors, immunomodulators (e.g., immunostimulants), immunotherapeutic agents, immune-based therapies, PI3K inhibitors, HIV antibodies, and bispecific antibodies, and "antibody-like" therapeutic proteins, and combinations thereof.

[0225] HIV combination drug In some embodiments, the antibodies or antigen-binding fragments described herein are combined with one, two, three, four, or more additional anti-HIV therapeutic agents. Examples of anti-HIV therapeutic agents that can be combined include ATRIPLA® (efavirenz, tenofovir disoproxil fumarate, and emtricitabine); COMPLERA® (EVIPLERA®; rilpivirine, tenofovir disoproxil fumarate, and emtricitabine); STRIBILD® (elvitegravir, cobicistat, tenofovir disoproxil fumarate, and emtricitabine); TRUVADA® (tenofovir disoproxil fumarate and emtricitabine; TDF+FTC); DESCCOVY® (tenofovir alafenamide and emtricitabine); ODEFSEY® (tenofovir alafenamide, emtricitabine, and rilpivirine); GENVOYA® (tenofovir alafenamide, emtricitabine, cobicistat, and elvitegravir); BIKTARVY (bictegravir + emtricitabine + tenofovir alafenamide), adefovir; adefovir dipivoxil; cobicistat; emtricitabine; tenofovir; tenofovir alafenamide and elvitegravir; tenofovir disoproxil; tenofovir disoproxil fumarate; tenofovir alafenamide; tenofovir alafenamide hemifumarate; TRIUMEQ® (dolutegravir, abacavir, and lamivudine); dolutegravir, abacavir sulfate, and lamivudine. Raltegravir; PEGylated raltegravir; raltegravir and lamivudine; maraviroc; tenofovir + emtricitabine + maraviroc, enfuvirtide; ALUVIA® (KALETRA®; lopinavir and ritonavir); COMBIVIR® (zidovudine and lamivudine; AZT+3TC); EPZICOM® (LIVEXA®; abacavir sulfate and lamivudine; ABC+3TC); TRIZIVIR® (abacavir sulfate, zidovudine, and lamivudine; ABC+AZT+3TC); atazanavir and cobicistat; atazanavir sulfate and cobicistat;Atazanavir Sulfate and Ritonavir; Darunavir; Darunavir and Cobicistat; Dolutegravir and Rilpivirine; Dolutegravir and Rilpivirine Hydrochloride; Dolutegravir, Abacavir Sulfate, and Lamivudine; Lamivudine, Nevirapine, and Zidovudine; Raltegravir and Lamivudine; Doravirine, Lamivudine, and Tenofovir Disoproxil Fumarate; Doravirine, Lamivudine, and Tenofovir Disoproxil; Dolutegravir + Lamivudine, Lamivudine + Abacavir + Zidovudine, Lamivudine + Abacavir, Lamivudine + Tenofovir Disoproxil Fumarate, Lamivudine + Zidovudine + Nevirapine, Lopinavir + Ritonavir, Lopinavir + Ritonavir + Abacavir + Lamivudine, Lopinavir + Ritonavir + Zidovudine + Lamivudine, Tenofovir + Lamivudine, and Tenofovir Disoproxil Fumarate + Emtricitabine + Rilpivirine Hydrochloride, Lopinavir, Ritonavir, Zidovudine and Lamivudine; Cabotegravir + Rilpivirine; Elvitegravir (Elsulfavirine; VM-1500; VM-1500A); Rilpivirine; Rilpivirine Hydrochloride; Atazanavir Sulfate and Cobicistat; Atazanavir and Cobicistat; Darunavir and Cobicistat; Atazanavir; Atazanavir Sulfate; Dolutegravir; Elvitegravir; Ritonavir; Atazanavir Sulfate and Ritonavir; Darunavir; Lamivudine; Prolastin; fosamprenavir; fosamprenavir calcium efavirenz; efavirenz, Lamivudine, and Emtricitabine; Etravirine; Nelfinavir; Nelfinavir Mesylate; Interferon; Didanosine; Stavudine; Indinavir; Indinavir Sulfate; Tenofovir and Lamivudine; Zidovudine; Nevirapine; Saquinavir; Saquinavir Mesylate; Aldesleukin; Zalcitabine; Tipranavir; Amprenavir; Delavirdine; Delavirdine Mesylate; Radha-108 (Receptol); Lamivudine and Tenofovir Disoproxil Fumarate; efavirenz, Lamivudine, and Tenofovir Disoproxil Fumarate; Phosphazide; Lamivudine, Nevirapine, and Zidovudine; Abacavir; and Abacavir Sulfate, are included, but not limited to these.;

[0226] Other HIV drugs Examples of other drugs for treating HIV that can be combined with the agent of the present disclosure include aspernigrin C, acemannan, alisporivir, BanLec, deferiprone, gamimune, metenkephalin, naltrexone, prolastin, REP 9, RPI-MN, VSSP, H1 virus, SB-728-T, 1,5-dicaffeoylquinic acid, rHIV7-shl-TAR-CCR5RZ, AAV-eCD4-Ig gene therapy, MazF gene therapy, BlockAide, bevirimat derivatives, ABX-464, AG-1105, APH-0812, bryostatin analogs, BIT-225, CYT-107, CS-TATI-1, fluoro-beta-D-arabinose nucleic acid (FANA)-modified antisense oligonucleotides, FX-101, griffithsin, HGTV-43, HPH-116, HS-10234, hydroxychloroquine, IMB-10035, IMO-3100, IND-02, JL-18008, LADAVRU, MK-1376, MK-2048, MK-4250, MK-8507, MK-8558, MK-8591 (islatravir), NOV-205, OB-002H, ODE-Bn-TFV, M1-TFV, PA-1050040 (PA-040), PC-707, PGN-007, QF-036, S-648414, SCY-635, SB-9200, SCB-719, TR-452, TEV-90110, TEV-90112, TEV-90111, TEV-90113, RN-18, DIACC-1010, Fasnall, Immuglo, 2-CLIPS peptide, HRF-4467, thrombospondin analogs, TBL-1004HI, VG-1177, xl-081, rfhSP-D, [18F]-MC-225, URMC-099-C, RES-529, and VIR-576.

[0227] HIV protease inhibitors In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV protease inhibitor. Examples of HIV protease inhibitors include amprenavir, atazanavir, brecanavir, darunavir, fosamprenavir, fosamprenavir calcium, indinavir, indinavir sulfate, lopinavir, nelfinavir, nelfinavir mesylate, ritonavir, saquinavir, saquinavir mesylate, tipranavir, AEBL-2, DG-17, GS-1156, TMB-657 (PPL-100), T-169, BL-008, MK-8122, TMB-607, GRL-02031, and TMC-310911.

[0228] HIV ribonuclease H inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV ribonuclease H inhibitor. An example of an HIV ribonuclease H inhibitor that can be combined is NSC-727447.

[0229] HIV Nef inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV Nef inhibitor. An example of an HIV Nef inhibitor that can be combined is FP-1.

[0230] HIV reverse transcriptase inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with non-nucleoside or non-nucleotide inhibitors. Examples of non-nucleoside or non-nucleotide inhibitors of reverse transcriptase for HIV include dapivirine, delavirdine, delavirdine mesylate, doravirine, efavirenz, etravirine, lentivudine, nevirapine, rilpivirine, ACC-007, ACC-008, AIC-292, F-18, KM-023, PC-1005, VM-1500A-LAI, PF-3450074, elsulfavirine (extended-release oral, for HIV infection), elsulfavirine (long-acting injectable nanosuspension, for HIV infection), and elsulfavirine (VM-1500).

[0231] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with HIV nucleoside or nucleotide inhibitors. Examples of HIV nucleoside or nucleotide inhibitors of reverse transcriptase include adefovir, adefovir dipivoxil, azidothymidine, emtricitabine, tenofovir, tenofovir alafenamide, tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir octadecyloxyethyl ester (AGX-1009), tenofovir disoproxil hemifumarate, VIDEX® and VIDEXEC® (zidovudine, ddl), abacavir, abacavir sulfate, alovudine, apricitabine, censavudine, zidovudine, elvucitabine, festinavir, fosalbutidine xylil, CMX-157, dapivirine, doravirine, etravirine, OCR-5753, tenofovir disoproxil orotate, tidoxil fozidibine, lamivudine, phosphazide, stavudine, zalcitabine, didoxuridine, lobapholvir etalafernamide (GS-9131), GS-9148, MK-8504, MK-8591, MK-858, and KP-1461.

[0232] HIV Integrase Inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV integrase inhibitor. Examples of HIV integrase inhibitors include elvitegravir, elvitegravir (sustained release microcapsules), curcumin, curcumin derivatives, cholic acid, cholic acid derivatives, 3,5-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid derivatives, aurintricarboxylic acid, aurintricarboxylic acid derivatives, caffeic acid phenethyl ester, caffeic acid phenethyl ester derivatives, tilorone, tilorone derivatives, quercetin, quercetin derivatives, raltegravir, PEGylated raltegravir, dolutegravir, JTK-351, bictegravir, AVX-15567, cabotegravir (long-acting injectable), diketoquinoline-4-1 derivatives, integrase-LEDGF inhibitors, ledgins, M-522, M-532, MK-0536, NSC-310217, NSC-371056, NSC-48240, NSC-642710, NSC-699171, NSC-699172, NSC-699173, NSC-699174, stilbene disulfonic acid, T-169, STP-0404, VM-3500, and cabotegravir.

[0233] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV non-catalytic site or allosteric integrase inhibitor (NCINI). Examples of HIV non-catalytic site or allosteric integrase inhibitors (NCINI) include CX-05045, CX-05168, and CX-14442.

[0234] HIV entry inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV entry inhibitor. Examples of HIV entry (fusion) inhibitors include AAR-501, LBT-5001, lenacapavir, CCR5 inhibitors, gp41 inhibitors, CD4 attachment inhibitors, gp120 inhibitors, gp160 inhibitors, and CXCR4 inhibitors.

[0235] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a CCR5 inhibitor. Examples of CCR5 inhibitors include apravirine, vicriviroc, maraviroc, maraviroc (long-acting injectable nanoemulsion), cenicriviroc, leronlimab (PRO-140), Adaptavir (RAP-101), nifeviroc (TD-0232), anti-GP120 / CD4 or CCR5 bispecific antibodies, B-07, MB-66, polypeptide C25P, TD-0680, thioraviroc, and vMIP (Haimipu).

[0236] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a CXCR4 inhibitor. Examples of CXCR4 inhibitors include plerixafor, ALT-1188, N15 peptide, and vMIP (Haimipu).

[0237] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a gp41 inhibitor. Examples of gp41 inhibitors include albuvirtide, enfuvirtide, Griffithsin (gp41 / gp120 / gp160 inhibitor), BMS-986197, enfuvirtide biobetter, enfuvirtide biosimilar, HIV-1 fusion inhibitor (P26-Bapc), ITV-1, ITV-2, ITV-3, ITV-4, CPT-31, Cl3hmAb, PIE-12 trimer, and sifuvirtide.

[0238] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a CD4 attachment inhibitor. Examples of CD4 attachment inhibitors include ibalizumab and CADA analogs.

[0239] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a gp120 inhibitor. Examples of gp120 inhibitors include anti-HIV microbicides, Radha-108 (receptor) 3B3-PE38, BanLec, bentonite-based nanomedicine, fostemsavir tromethamine, IQP-0831, VVX-004, and BMS-663068.

[0240] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a gp160 inhibitor. Examples of gp160 inhibitors that can be combined include phanquinoline.

[0241] HIV maturation inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV maturation inhibitor. Examples of HIV maturation inhibitors include BMS-955176, GSK-3640254, and GSK-2838232.

[0242] Latency antagonist In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV latency reversing agent. Examples of latency reversing agents that can be combined with one or more of the multispecific antigen-binding molecules described herein include IL-15 receptor agonists (e.g., ALT-803; interleukin-15 / Fc fusion proteins (e.g., XmAb24306)); recombinant interleukin-15 (e.g., AM0015, NIZ-985); pegylated IL-15 (e.g., NKTR-255); toll-like receptor (TLR) agonists (including TLR7 agonists such as GS-9620 and TLR8 agonists such as GS-9688), histone deacetylase (HDAC) inhibitors, proteasome inhibitors such as bortezomib, protein kinase C (PKC) activators, Smyd2 inhibitors, BET-bromodomain 4 (BRD4) inhibitors, ionomycin, IAP antagonists (inhibitors of apoptosis proteins such as APG-1387, LBW-242), SMAC mimetics (such as ciapavir, TL32711, LCL161, GDC-0917, HGS1029, AT-406), dBeQ-1143, PMA, SAHA (suberanilohydroxamic acid, or suberoyl, anilide, and hydroxamic acid), NIZ-985, IL-15 modulating antibodies (including IL-15, IL-15 fusion proteins and IL-15 receptor agonists such as ALT-803), JQ1, disulfiram, amphotericin B, and ubiquitin inhibitors such as largazole analogs, APH-0812, and GSK-343. Examples of HDAC inhibitors include romidepsin, vorinostat, and panobinostat. Examples of PKC activators include indolactam, prostratin, ingenol B, and DAG-lactone.

[0243] Toll-like receptor (TLR) agonist In some embodiments, the antibodies or antigen-binding fragments described herein are combined with agonists of toll-like receptors (TLRs), such as agonists of TLR1 (NCBI Gene ID: 7096), TLR2 (NCBI Gene ID: 7097), TLR3 (NCBI Gene ID: 7098), TLR4 (NCBI Gene ID: 7099), TLR5 (NCBI Gene ID: 7100), TLR6 (NCBI Gene ID: 10333), TLR7 (NCBI Gene ID: 51284), TLR8 (NCBI Gene ID: 51311), TLR9 (NCBI Gene ID: 54106), and / or TLR10 (NCBI Gene ID: 81793).Exemplary TLR7 agonists that can be co-administered or combined with one or more multispecific antigen-binding molecules described herein include AL-034, DSP-0509, GS-9620 (bexarotene), bexarotene analogs, LHC-165, TMX-101 (imiquimod), GSK-2245035, resiquimod, DSR-6434, DSP-3025, IMO-4200, MCT-465, MEDI-9197, 3M-051, SB-9922, 3M-052, Limtop, TMX-30X, TMX-202, RG-7863, RG-7854, RG-7795, and the compounds disclosed in US20100143301 (Gilead Sciences), US20110098248 (Gilead Sciences), and US20090047249 (Gilead Sciences), US20140045849 (Janssen), US20140073642 (Janssen), WO2014 / 056953 (Janssen), WO2014 / 076221 (Janssen), WO2014 / 128189 (Janssen), US20140350031 (Janssen), WO2014 / 023813 (Janssen), US20080234251 (Array Biopharma), US20080306050 (Array Biopharma), US20100029585 (Ventirx Pharma), US20110092485 (Ventirx Pharma), US20110118235 (Ventirx Pharma), US20120082658 (Ventirx Pharma), US20120219615 (Ventirx Pharma), US20140066432 (Ventirx Pharma), US20140088085 (Ventirx Pharma), US20140275167 (Novira Therapeutics), and US20130251673 (Novira Therapeutics), but are not limited thereto. TLR7 / TLR8 agonists that can be co-administered are NKTR-262, telratolimod, and BDB-001.Examples of TLR8 agonists that can be co-administered or combined with one or more multispecific antigen-binding molecules described herein include E-6887, IMO-4200, IMO-8400, IMO-9200, MCT-465, MEDI-9197, motolimod, resiquimod, GS-9688, VTX-1463, VTX-763, 3M-051, 3M-052, and the compounds described in US20140045849 (Janssen), US20140073642 (Janssen), WO2014 / 056953 (Janssen), WO2014 / 076221 (Janssen), WO2014 / 128189 (Janssen), US20140350031 (Janssen), WO2014 / 023813 (Janssen), US20080234251 (Array Biopharma), US20080306050 (Array Biopharma), US20100029585 (Ventirx Pharma), US20110092485 (Ventirx Pharma), US20110118235 (Ventirx Pharma), US20120082658 (Ventirx Pharma), US20120219615 (Ventirx Pharma), US20140066432 (Ventirx Pharma), US20140088085 (Ventirx Pharma), US20140275167 (Novira Therapeutics), and US20130251673 (Novira Therapeutics), but are not limited thereto. Examples of TLR9 agonists that can be co-administered include AST-008, cobitolimod, CMP-001, IMO-2055, IMO-2125, ritenimod, MGN-1601, BB-001, BB-006, IMO-3100, IMO-8400, IR-103, IMO-9200, agatolimod, DIMS-9054, DV-1079, DV-1179, AZD-1419, lefitolimod (MGN-1703), CYT-003, CYT-003-QbG10, tilsotolimod, and PUL-042, but are not limited thereto.Examples of TLR3 agonists include lintatrimod, poly ICLC, RIBOXXON®, Apoxxim, RIBOXXIM®, IPH-33, MCT-465, MCT-475, and ND-1.1. Examples of TLR4 agonists include G-100 and GSK-1795091.

[0244] Histone deacetylase (HDAC) inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an inhibitor of histone deacetylase, such as histone deacetylase 1, histone deacetylase 9 (HDAC9, HD7, HD7b, HD9, HDAC, HDAC7, HDAC7B, HDAC9B, HDAC9FL, HDRP, MITR; Gene ID: 9734). Examples of HDAC inhibitors include, but are not limited to, abexinostat, ACY-241, AR-42, BEBT-908, belinostat, CKD-581, CS-055 (HBI-8000), CT-101, CUDC-907 (fimepinostat), entinostat, givinostat, mocetinostat, panobinostat, pracinostat, xenoSTAT (JNJ-26481585), resminostat, ricolinostat, romidepsin, SHP-141, TMB-ADC, valproic acid (VAL-001), vorinostat, tinostamustine, remetinostat, and entinostat.

[0245] Cyclin-dependent kinase (CDK) inhibitor or antagonist In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an inhibitor or antagonist of a cyclin-dependent kinase (CDK), such as cyclin-dependent kinase 4 (CDK4; NCBI Gene ID: 1019), cyclin-dependent kinase 6 (CDK6; NCBI Gene ID: 1021), cyclin-dependent kinase 9 (CDK9; NCBI Gene ID: 1025). In some embodiments, the CDK4 / CDK6 / CDK9 inhibitor or antagonist is selected from the group consisting of VS2-370.

[0246] Stimulator of interferon genes (STING) agonist In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a stimulator of interferon genes (STING). In some embodiments, the STING receptor agonist or activator is selected from the group consisting of ADU-S100 (MIW-815), SB-11285, MK-1454, SR-8291, AdVCA0848, GSK-532, SYN-STING, MSA-1, SR-8291, 5,6-dimethylxanthenone-4-acetic acid (DMXAA), cyclic GAMP (cGAMP), and cyclic di-AMP.

[0247] RIG-I agonist In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an agonist of DExD / H-box helicase 58 (DDX58; also known as RIG-I, RIG1, RIGI, RLR-1, SGMRT2; NCBI Gene ID: 23586). In some embodiments, the agents described herein are combined with an RIG-1 modulator such as RGT-100, or an NOD2 modulator such as SB-9200 (also known as GS 9992; Inaligivir), and IR-103. An exemplary RIG-I agonist is KIN1148, as described by Hemann, et al., J Immunol May 1, 2016, 196 (1 Supplement) 76.1. Additional RIG-I agonists are described, for example, in Elion, et al., Cancer Res. (2018) 78(21):6183-6195; and Liu, et al., J Virol. (2016) 90(20):9406-19. RIG-I agonists are commercially available, for example, from Invivogen (invivogen.com).

[0248] LAG-3 inhibitor and TIM-3 inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an anti-TIM-3 (also known as hepatitis A virus cellular receptor 2 antibody (HAVCR2; NCBI Gene ID: 84868)) such as TSR-022, LY-3321367, MBG-453, INCAGN-2390. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an anti-LAG-3 (lymphocyte activation) (NCBI Gene ID: 3902) antibody such as Relatlimab (ONO-4482), LAG-525, MK-4280, REGN-3767, INCAGN2385.

[0249] Capsid inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a capsid inhibitor. Examples of capsid inhibitors that can be combined with the agents of the present disclosure include capsid polymerization inhibitors or capsid disrupting compounds, HIV nucleocapsid p7 (NCp7) inhibitors such as azodicarboxamide, HIV p24 capsid protein inhibitors, GS-6207 (lenacapavir), GS-CA1, AVI-621, AVI-101, AVI-201, AVI-301, and the AVI-CAN1-15 series, PF-3450074, and the compounds described in International Patent Publication No. WO2019 / 087016.

[0250] Immune-based therapies In some embodiments, the antibodies or antigen-binding fragments described herein are combined with immune-based therapies. Examples of immune-based therapies include toll-like receptor (TLR) modulators such as TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TLR12, TLR13; programmed cell death protein 1 (PD-1) modulators; programmed death ligand 1 (PD-L1) modulators; IL-15 modulators (e.g., IL-15 receptor agonists (e.g., ALT-803; interleukin-15 / Fc fusion proteins (e.g., XmAb24306)); recombinant interleukin-15 (e.g., AM0015, NIZ-985); pegylated IL-15 (e.g., NKTR-255)); DermaVir; interleukin-7; plaquenil (hydroxychloroquine); proleukin (aldesleukin, IL-2); interferon alpha; interferon alpha-2b; interferon alpha-n3; pegylated interferon alpha; interferon gamma; hydroxyurea; mycophenolate mofetil (MPA) and its ester derivative mycophenolate mofetil (MMF); ribavirin; polymeric polyethyleneimine (PEI); gepon; IL-12; WF-10; VGV-1; MOR-22; BMS-936559; CYT-107, norm interferon, pegylated interferon alpha-2a, pegylated interferon alpha-2b, RPI-MN, STING modulators, RIG-I modulators, NOD2 modulators, SB-9200, and IR-103.

[0251] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a TLR agonist. Examples of TLR agonists include, but are not limited to, besatollimod (GS-9620), refitolimod, tildrakizumab, lintetimod, DSP-0509, AL-034, G-100, cobitolimod, AST-008, motolimod, GSK-1795091, GSK-2245035, VTX-1463, GS-9688, LHC-165, BDB-001, RG-7854, telratolimod.

[0252] CD47 targeting agent In various embodiments, the antibodies or antigen-binding fragments described herein are combined with an agent that interferes with the binding of CD47 to SIRPα, such as an agent that targets CD47 or an agent that targets SIRPα. In various embodiments, the antibodies or antigen-binding fragments described herein are combined with an inhibitor of CD47 (IAP, MER6, OA3; NCBI Gene ID: 961). Examples of CD47 inhibitors include, but are not limited to, anti-CD47 mAb (Vx-1004), anti-human CD47 mAb (CNTO-7108), CC-90002, CC-90002-ST-001, humanized anti-CD47 antibody (Hu5F9-G4; magrolimab), NI-1701, NI-1801, RCT-1938, and TTI-621. In some embodiments, the CD47 inhibitor is magrolimab.

[0253] Immune checkpoint receptor protein modulator In various embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more blockers or inhibitors of one or more inhibitory immune checkpoint proteins or receptors and / or one or more stimulants, activators or agonists of one or more stimulatory immune checkpoint proteins or receptors. Blocking or inhibiting an inhibitory immune checkpoint can actively regulate the activation of T cells or NK cells and inhibit immune evasion by infected cells. Activation or stimulation of a stimulatory immune checkpoint can enhance the effect of immune checkpoint inhibitors in infectious treatment. In various embodiments, the immune checkpoint protein or receptor regulates the T cell response (e.g., reviewed in Xu, et al., J Exp Clin Cancer Res. (2018) 37:110). In various embodiments, the immune checkpoint protein or receptor regulates the NK cell response (e.g., reviewed in Davis, et al., Semin Immunol. (2017) 31:64-75, and Chiossone, et al., Nat Rev Immunol. (2018) 18(11):671-688).

[0254] Examples of immune checkpoint proteins or receptors that can be combined with the antibodies or antigen-binding fragments described herein include CD27, CD70; CD40, CD40LG; CD47, CD48 (SLAMF2), Transmembrane and Immunoglobulin Domain Containing 2 (TMIGD2, CD28H), CD84 (LY9B, SLAMF5), CD96, CD160, MS4A1 (CD20), CD244 (SLAMF4); CD276 (B7H3); V-set domain containing T cell activation inhibitor 1 (VTCN1, B7H4); V-set immunoregulatory receptor (VSIR, B7H5, VISTA); Immunoglobulin Superfamily Member 11 (IGSF11, VSIG3); Natural Killer Cell Cytotoxicity Receptor 3 Ligand 1 (NCR3LG1, B7H6); HERV-H LTR Associated 2 (HHLA2, B7H7); Inducible T cell co-stimulator (ICOS, CD278); Inducible T cell co-stimulatory ligand (ICOSLG, B7H2); TNF Receptor Superfamily Member 4 (TNFRSF4, OX40); TNF Superfamily Member 4 (TNFSF4, OX40L); TNFRSF8 (CD30), TNFSF8 (CD30L); TNFRSF10A (CD261, DR4, TRAILR1), TNFRSF9 (CD137), TNFSF9 (CD137L); TNFRSF10B (CD262, DR5, TRAILR2), TNFRSF10 (TRAIL); TNFRSF14 (HVEM, CD270), TNFSF14 (HVEML); CD272 (B and T lymphocyte associated (BTLA)); TNFRSF17 (BCMA, CD269), TNFSF13B (BAFF); TNFRSF18 (GITR), TNFSF18 (GITRL); MHC Class I polypeptide-related sequence A (MICA); MHC Class I polypeptide-related sequence B (MICB); CD274 (CD274, PDL1, PD-L1); Programmed cell death 1 (PDCD1, PD1, PD-1); Cytotoxic T lymphocyte-associated protein 4 (CTLA4, CD152); CD80 (B7-1), CD28; Necltin cell adhesion molecule 2 (NECTIN2, CD112); CD226 (DNAM-1); Polyomavirus receptor (PVR) cell adhesion molecule (PVR, CD155); PVR-related immunoglobulin domain containing (PVRIG, CD112R);T cell immunoreceptor with Ig and ITIM domains (TIGIT); T cell immunoglobulin and mucin domain 4 (TIMD4; TIM4) including 4; hepatitis A virus cellular receptor 2 (HAVCR2, TIMD3, TIM3); galectin 9 (LGALS9); lymphocyte activation 3 (LAG3, CD223); signaling lymphocyte activation molecule family member 1 (SLAMF1, SLAM, CD150); lymphocyte antigen 9 (LY9, CD229, SLAMF3); SLAM family member 6 (SLAMF6, CD352); SLAM family member 7 (SLAMF7, CD319); UL16 binding protein 1 (ULBP1); UL16 binding protein 2 (ULBP2); UL16 binding protein 3 (ULBP3); retinoic acid early transcript 1E (RAET1E; ULBP4); retinoic acid early transcript 1G (RAET1G; ULBP5); retinoic acid early transcript 1L (RAET1L; ULBP6); lymphocyte activation 3 (CD223); killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 1 (KIR, CD158E1); killer cell lectin like receptor C1 (KLRC1, NKG2A, CD159A); killer cell lectin like receptor K1 (KLRK1, NKG2D, CD314); killer cell lectin like receptor C2 (KLRC2, CD159c, NKG2C); killer cell lectin like receptor C3 (KLRC3, NKG2E); killer cell lectin like receptor C4 (KLRC4, NKG2F); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3); killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 1 (KIR3DL1); killer cell lectin like receptor D1 (KLRD1); and SLAM family member 7 (SLAMF7), including but not limited to these.;

[0255] In various embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more blockers or inhibitors of one or more T cell inhibitory immune checkpoint proteins or receptors. Exemplary T cell inhibitory immune checkpoint proteins or receptors include CD274 (CD274, PDL1, PD-L1); programmed cell death 1 ligand 2 (PDCD1LG2, PD-L2, CD273); programmed cell death 1 (PDCD1, PD1, PD-1); cytotoxic T lymphocyte-associated protein 4 (CTLA4, CD152); CD276 (B7H3); V-set domain containing T cell activation inhibitor 1 (VTCN1, B7H4); V-set immune regulatory receptor (VSIR, B7H5, VISTA); immunoglobulin superfamily member 11 (IGSF11, VSIG3); TNFRSF14 (HVEM, CD270), TNFSF14 (HVEML); CD272 (B and T lymphocyte associated (BTLA)); PVR-related immunoglobulin domain containing (PVRIG, CD112R); T cell immunoreceptor with Ig domain and ITIM domain (TIGIT); lymphocyte activation 3 (LAG3, CD223); hepatitis A virus cellular receptor 2 (HAVCR2, TIMD3, TIM3); galectin 9 (LGALS9); killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR, CD158E1); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3); and killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR3DL1), but are not limited thereto. In various embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more agonists or activators of one or more T cell stimulatory immune checkpoint proteins or receptors.Exemplary T cell-stimulatory immune checkpoint proteins or receptors include, but are not limited to, CD27, CD70; CD40, CD40LG; inducible T cell co-stimulatory device (ICOS, CD278); inducible T cell co-stimulatory ligand (ICOSLG, B7H2); TNF receptor superfamily member 4 (TNFRSF4, OX40); TNF superfamily member 4 (TNFSF4, OX40L); TNFRSF9 (CD137), TNFSF9 (CD137L); TNFRSF18 (GITR), TNFSF18 (GITRL); CD80 (B7-1), CD28; nectin cell adhesion molecule 2 (NECTIN2, CD112); CD226 (DNAM-1); CD244 (2B4, SLAMF4), poliovirus receptor (PVR) cell adhesion molecule (PVR, CD155). See, for example, Xu, et al., J Exp Clin Cancer Res. (2018) 37:110.

[0256] In various embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more blockers or inhibitors of one or more NK cell inhibitory immune checkpoint proteins or receptors. Exemplary NK cell inhibitory immune checkpoint proteins or receptors include killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR, CD158E1); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3); killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR3DL1); killer cell lectin-like receptor C1 (KLRC1, NKG2A, CD159A); and killer cell lectin-like receptor D1 (KLRD1, CD94), but are not limited thereto. In various embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more agonists or activators of one or more NK cell stimulatory immune checkpoint proteins or receptors. Exemplary NK cell stimulatory immune checkpoint proteins or receptors include CD16, CD226 (DNAM-1); CD244 (2B4, SLAMF4); killer cell lectin-like receptor K1 (KLRK1, NKG2D, CD314); SLAM family member 7 (SLAMF7), but are not limited thereto. See, for example, Davis, et al., Semin Immunol. (2017) 31:64-75; Fang, et al., Semin Immunol. (2017) 31:37-54; and Chiossone, et al., Nat Rev Immunol. (2018) 18(11):671-688.

[0257] In some embodiments, the one or more immune checkpoint inhibitors include proteinaceous (e.g., antibody or fragment thereof, or antibody mimetic) inhibitors of PD-L1 (CD274), PD-1 (PDCD1) or CTLA4. In some embodiments, the one or more immune checkpoint inhibitors include small molecule organic inhibitors of PD-L1 (CD274), PD-1 (PDCD1), or CTLA4.

[0258] Examples of inhibitors of CTLA4 that can be co-administered include, but are not limited to, ipilimumab, tremelimumab, BMS-986218, AGEN1181, AGEN1884, BMS-986249, MK-1308, REGN-4659, ADU-1604, CS-1002, BCD-145, APL-509, JS-007, BA-3071, ONC-392, AGEN-2041, JHL-1155, KN-044, CG-0161, ATOR-1144, PBI-5D3H5, BPI-002, and the bispecific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), MEDI-5752 (CTLA4 / PD-1), XmAb-20717 (PD-1 / CTLA4), and AK-104 (CTLA4 / PD-1).

[0259] Examples of inhibitors of PD-L1 (CD274) or PD-1 (PDCD1) that can be co-administered include pembrolizumab, nivolumab, semipramab, pidilizumab, AMP-224, MEDI0680 (AMP-514), spartalizumab, atezolizumab, avelumab, durvalumab, BMS-936559, CK-301, PF-06801591, BGB-A317 (tislelizumab), GLS-010 (WBP-3055), AK-103 (HX-008), AK-105, CS-1003, HLX-10, MGA-012, BI-754091, AGEN-2034, JS-001 (toripalimab), JNJ-63723283, genolimzumab (CBT-501), LZM-009, BCD-100, LY-3300054, SHR-1201, SHR-1210 (camrelizumab), Sym-021, ABBV-181 (buzigali mab), PD1-PIK, BAT-1306, (MSB0010718C), CX-072, CBT-502, TSR-042 (dostarlimab), MSB-2311, JTX-4014, BGB-A333, SHR-1316, CS-1001, WBP-3155, KN-035, IBI-308 (sintilimab), HLX-20, KL-A167, STI-A1014, STI-A1015 (IMC-001), BCD-135, FAZ-053, TQB-2450, MDX1105-01, GS-4224, GS-4416, INCB086550, MAX10181, and the bispecific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-013 (PD-1 / LAG-3), FS-118 (LAG-3 / PD-L1) MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), MEDI-5752 (CTLA4 / PD-1), RO-7121661 (PD-1 / TIM-3), XmAb-20717 (PD-1 / CTLA4), AK-104 (CTLA4 / PD-1), M7824 (PD-L1 / TGFβ-EC domain), CA-170 (PD-L1 / VISTA), CDX-527 (CD27 / PD-L1), LY-3415244 (TIM3 / PDL1), and INBRX-105 (4-1BB / PDL1), but are not limited to these.

[0260] In some embodiments, the small molecule inhibitor of CD274 or PDCD1 is selected from the group consisting of GS-4224, GS-4416, INCB086550, and MAX10181. In some embodiments, the small molecule inhibitor of CTLA4 includes BPI-002.

[0261] In various embodiments, the antibodies or antigen-binding fragments described herein are combined with anti-TIGIT antibodies such as etigilimab, BMS-986207, tirigolumab (also known as MTIG-7192A; RG-6058; RO 7092284), AGEN1307, AGEN1327, AGEN1777, COM-902, IBI-939, AB154, MG1131, and EOS884448 (EOS-448).

[0262] TNF receptor superfamily (TNFRSF) member agonist or activator In various embodiments, the antibodies or antigen-binding fragments described herein are agonists of one or more members of the TNF receptor superfamily (TNFRSF), e.g., agonists of one or more TNFRSF1A (NCBI Gene ID: 7132), TNFRSF1B (NCBI Gene ID: 7133), TNFRSF4 (OX40, CD134; NCBI Gene ID: 7293), TNFRSF5 (CD40; NCBI Gene ID: 958), TNFRSF6 (FAS, NCBI Gene ID: 355), TNFRSF7 (CD27, NCBI Gene ID: 939), TNFRSF8 (CD30, NCBI Gene ID: 943), TNFRSF9 (4-1BB, CD137, NCBI Gene ID: 3604), TNFRSF10A (CD261, DR4, TRAILR1, NCBI Gene ID: 8797), TNFRSF10B (CD262, DR5, TRAILR2, NCBI Gene ID: 8795), TNFRSF10C (CD263, TRAILR3, NCBI Gene ID: 8794), TNFRSF10D (CD264, TRAILR4, NCBI Gene ID: 8793), TNFRSF11A (CD265, RANK, NCBI Gene ID: 8792), TNFRSF11B (NCBI Gene ID: 4982), TNFRSF12A (CD266, NCBI Gene ID: 51330), TNFRSF13B (CD267, NCBI Gene ID: 23495), TNFRSF13C (CD268, NCBI Gene ID: 115650), TNFRSF16 (NGFR, CD271, NCBI Gene ID: 4804), TNFRSF17 (BCMA, CD269, NCBI Gene ID: 608), TNFRSF18 (GITR, CD357, NCBI Gene ID: 8784), TNFRSF19 (NCBI Gene ID: 55504), TNFRSF21 (CD358, DR6, NCBI Gene ID: 27242), and TNFRSF25 (DR3, NCBI Gene ID: 8718).

[0263] Examples of anti-TNFRSF4 (OX40) antibodies that can be co-administered include, but are not limited to, MEDI6469, MEDI6383, MEDI0562 (tabalixizumab), MOXR0916, PF-04518600, RG-7888, GSK-3174998, INCAGN1949, BMS-986178, GBR-8383, ABBV-368, and those described in WO2016179517, WO2017096179, WO2017096182, WO2017096281, and WO2018089628.

[0264] Examples of anti-TNFRSF5 (CD40) antibodies that can be co-administered include, but are not limited to, RG7876, SEA-CD40, APX-005M, and ABBV-428.

[0265] In some embodiments, the anti-TNFRSF7 (CD27) antibody balilumab (CDX-1127) is co-administered.

[0266] Examples of anti-TNFRSF9 (4-1BB, CD137) antibodies that can be co-administered include, but are not limited to, urelumab, utomilumab (PF-05082566), AGEN2373, and ADG-106.

[0267] Examples of anti-TNFRSF18 (GITR) antibodies that can be co-administered include, but are not limited to, MEDI1873, FPA-154, INCAGN-1876, TRX-518, BMS-986156, MK-1248, GWN-323, and those described in WO2017096179, WO2017096276, WO2017096189, and WO2018089628. In some embodiments, an antibody or fragment thereof that simultaneously targets TNFRSF4 (OX40) and TNFRSF18 (GITR) is co-administered. Such antibodies are described, for example, in WO2017096179 and WO2018089628.

[0268] Interleukin receptor agonist In some embodiments, the antibodies or antigen-binding fragments described herein are interleukin receptor agonists such as IL-2, IL-7, IL-15, IL-10, IL-12 agonists; examples of IL-2 receptor agonists such as proleukin (aldesleukin, IL-2); pegylated IL-2 (e.g., NKTR-214); modified variants of IL-2 (e.g., THOR-707), bempegaldesleukin, AIC-284, ALKS-4230, CUI-101, Neo-2 / 15; IL-15 receptor agonists such as ALT-803, NKTR-255, hetIL-15, interleukin-15 / Fc fusion protein, AM-0015, NIZ-985, SO-C101, IL-15 syntocin (pegylated IL-15), P-22339, and IL-15-PD-1 fusion protein N-809; and in combination with; an example of IL-7 is CYT-107.

[0269] Examples of additional interleukin receptor agonists that can be combined with the antibodies or antigen-binding fragments described herein include interferon alpha; interferon alpha-2b; interferon alpha-n3; pegylated interferon alpha; interferon gamma; Flt3 agonists such as CDX-301; gepon; norm interferon, peginterferon alpha-2a, peginterferon alpha-2b, RPI-MN.

[0270] Bispecific and trispecific natural killer (NK)-cell engagement In various embodiments, the antibodies or antigen-binding fragments described herein are combined with bispecific NK cell engagers (BiKEs) or trispecific NK cell engagers (TriKEs) (e.g., lacking an Fc), or bispecific antibodies (e.g., having an Fc) against NK cell activating receptors such as CD16A, C-type lectin receptors (CD94 / NKG2C, NKG2D, NKG2E / H, and NKG2F), natural killer cell cytotoxicity receptors (NKp30, NKp44, and NKp46), killer cell C-type lectin-like receptors (NKp65, NKp80), Fc receptors FcγR (which mediate antibody-dependent cell cytotoxicity), SLAM family receptors (2B4, SLAM6, SLAM7, etc.), killer cell immunoglobulin-like receptors (KIR) (KIR-2DS and KIR-3DS), DNAM-1, and CD137 (4-1BB). Exemplary anti-CD16 bispecific antibodies, BiKEs, or TriKEs that can be co-administered include AFM26 (BCMA / CD16A) and AFM-13 (CD16 / CD30). Optionally, the anti-CD16 binding bispecific molecule may or may not have an Fc. BiKEs and TriKEs are described, for example, in Felices, et al., Methods Mol Biol. (2016) 1441:333-346; Fang, et al., Semin Immunol. (2017) 31:37-54. Examples of trispecific NK cell engagers (TRiKEs) include OXS-3550 and CD16-IL-15-B7H3 TriKe.

[0271] Phosphatidylinositol 3-kinase (PI3K) inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a PI3K inhibitor. Examples of PI3K inhibitors include idelalisib, alpelisib, buparlisib, CAI orotate, copanlisib, duvelisib, gedatolisib, neratinib, panpulisib, perifosine, pictilisib, pilaralisib, pucritinib mesylate, rigosertib, rigosertib sodium, sonolisib, taselisib, AMG-319, AZD-8186, BAY-1082439, CLR-1401, CLR-457, CUDC-907, DS-7423, EN-3342, GSK-2126458, GSK-2269577, GSK -2636771, INCB-040093, LY-3023414, MLN-1117, PQR-309, RG-7666, RP-6530, RV-1729, SAR-245409, SAR-260301, SF-1126, TGR-1202, UCB-5857, VS-5584, XL-765, and ZSTK-474.

[0272] Alpha-4 / Beta-7 antagonist In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an alpha-4 / beta-7 antagonist. Examples of integrin α-4 / β-7 antagonists include PTG-100, TRK-170, abrilumab, etrolizumab, carotegrast methyl, and vedolizumab.

[0273] Pharmacokinetic enhancer In some embodiments, the antibodies or antigen-binding fragments described herein are combined with a pharmacokinetic enhancer. Examples of pharmacokinetic enhancers include cobicistat and ritonavir.

[0274] Additional therapeutic agent Examples of additional therapeutic agents include the compounds disclosed in WO 2004 / 096286 (Gilead Sciences); WO 2006 / 015261 (Gilead Sciences); WO 2006 / 110157 (Gilead Sciences); WO 2012 / 003497 (Gilead Sciences); WO 2012 / 003498 (Gilead Sciences); WO 2012 / 145728 (Gilead Sciences); WO 2013 / 006738 (Gilead Sciences); WO 2013 / 159064 (Gilead Sciences); WO 2014 / 100323 (Gilead Sciences), US 2013 / 0165489 (University of Pennsylvania), US 2014 / 0221378 (Japan Tobacco), US 2014 / 0221380 (Japan Tobacco); WO 2009 / 062285 (Boehringer Ingelheim); WO 2010 / 130034 (Boehringer Ingelheim); WO 2013 / 006792 (Pharma Resources), US 20140221356 (Gilead Sciences), US 20100143301 (Gilead Sciences), and WO 2013 / 091096 (Boehringer Ingelheim).

[0275] HIV combination therapy In certain embodiments, the antibodies or antigen-binding fragments described herein are ATRIPLA® (efavirenz, tenofovir disoproxil fumarate, and emtricitabine); COMPLERA® (EVIPLERA®; rilpivirine, tenofovir disoproxil fumarate, and emtricitabine); STRIBILD® (elvitegravir, cobicistat, tenofovir disoproxil fumarate, and emtricitabine); TRUVADA® (tenofovir disoproxil fumarate and emtricitabine; TDF+FTC); DESCCOVY® (tenofovir alafenamide and emtricitabine); ODEFSEY® (tenofovir alafenamide, emtricitabine, and rilpivirine); GENVOYA® (tenofovir alafenamide, emtricitabine, cobicistat, and elvitegravir); adefovir; adefovir dipivoxil; cobicistat; emtricitabine; tenofovir; tenofovir disoproxil; tenofovir disoproxil fumarate; tenofovir alafenamide; tenofovir alafenamide hemifumarate; TRIUMEQ® (dolutegravir, abacavir, and lamivudine); dolutegravir, abacavir sulfate, and lamivudine.Raltegravir; Raltegravir and Lamivudine; Maraviroc; Enfuvirtide; ALUVIA® (KALETRA®; Ritonavir and Lopinavir); COMBIVIR® (Zidovudine and Lamivudine; AZT + 3TC); EPZICOM® (LIVEXA®; Abacavir Sulfate and Lamivudine; ABC + 3TC); TRIZIVIR® (Abacavir Sulfate, Zidovudine, and Lamivudine; ABC + AZT + 3TC); Rilpivirine; Rilpivirine Hydrochloride; Atazanavir Sulfate and Cobicistat; Atazanavir and Cobicistat; Darunavir and Cobicistat; Atazanavir; Atazanavir Sulfate; Dolutegravir; Elvitegravir; Ritonavir; Atazanavir Sulfate and Ritonavir; Darunavir; Lamivudine; Prolactin; Fosamprenavir; Fosamprenavir Calcium Efavirenz; Etravirine; Nelfinavir; Nelfinavir Mesylate; Interferon; Didanosine; Stavudine; Indinavir; Indinavir Sulfate; Tenofovir and Lamivudine; Zidovudine; Nevirapine; Saquinavir; Saquinavir Mesylate; Aldesleukin; Zalcitabine; Tipranavir; Amprenavir; Delavirdine; Delavirdine Mesylate; Radha-108 (Receptor); Lamivudine and Tenofovir Disoproxil Fumarate; Efavirenz, Lamivudine, and Tenofovir Disoproxil Fumarate; Phosphazide; Lamivudine, Nevirapine, and Zidovudine; Abacavir; and Abacavir Sulfate, is combined with one, two, three, four or more additional therapeutic agents selected from the group consisting of

[0276] It will be understood by those skilled in the art that the additional therapeutic agents described above may be included in two or more of the above classes.

[0277] In certain embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV nucleoside or nucleotide inhibitor of reverse transcriptase and an HIV non-nucleoside inhibitor of reverse transcriptase. In other certain embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV nucleoside or nucleotide inhibitor of reverse transcriptase, and an HIV protease inhibitor compound. In further embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV nucleoside or nucleotide inhibitor of reverse transcriptase, an HIV non-nucleoside inhibitor of reverse transcriptase, and a pharmacokinetic enhancer. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with at least one HIV nucleoside inhibitor of reverse transcriptase, an integrase inhibitor, and a pharmacokinetic enhancer. In other embodiments, the antibodies or antigen-binding fragments described herein are combined with two HIV nucleoside or nucleotide inhibitors of reverse transcriptase.

[0278] In certain embodiments, the antibodies or antigen-binding fragments described herein are combined with abacavir sulfate, tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, tenofovir alafenamide, or tenofovir alafenamide hemifumarate.

[0279] In certain embodiments, the antibodies or antigen-binding fragments described herein are combined with tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide, or tenofovir alafenamide hemifumarate.

[0280] In certain embodiments, the antibodies or antigen-binding fragments described herein are combined with a first additional therapeutic agent selected from the group consisting of abacavir sulfate, tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide, and tenofovir alafenamide hemifumarate, and a second additional therapeutic agent selected from the group consisting of emtricitabine and lamivudine.

[0281] In certain embodiments, the antibodies or antigen-binding fragments described herein are combined with a first additional therapeutic agent selected from the group consisting of tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide, and tenofovir alafenamide hemifumarate, and a second additional therapeutic agent, wherein the second additional therapeutic agent is emtricitabine.

[0282] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more additional therapeutic agents at a therapeutically effective dosage of the antibody or antigen-binding fragment in the range of, for example, 1 mg to 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 1000 mg, or 1500 mg. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more additional therapeutic agents at a therapeutically effective dosage of the antibody or antigen-binding fragment in the range of, for example, about 0.1 mg / kg to about 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 8 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, or 50 mg / kg. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with one or more additional therapeutic agents at a therapeutically effective dosage of the antibody or antigen-binding fragment in the range of, for example, about 5 mg to about 10 mg, 20 mg, 25 mg, 50 mg, 100 mg, 125 mg, 150 mg, 250 mg, 300 mg, 500 mg, 1000 mg, or 1500 mg.

[0283] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with 5 to 30 mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide, and 200 mg of emtricitabine. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with 5 to 10, 5 to 15, 5 to 20, 5 to 25, 25 to 30, 20 to 30, 15 to 30, or 10 to 30 mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide, and 200 mg of emtricitabine. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with 10 mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide, and 200 mg of emtricitabine. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with 25 mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide, and 200 mg of emtricitabine. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with the agents provided herein at any dosage of the antibody or antigen-binding fragment (e.g., 1 mg to 500 mg of the antibody or antigen-binding fragment as described herein) such that each combination of dosages is listed specifically and individually, in the same manner as the combinations of dosages are specifically and individually listed above.

[0284] In some embodiments, the antibodies or antigen-binding fragments described herein are combined with 200-400 mg of tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil, and 200 mg of emtricitabine. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with 200-250, 200-300, 200-350, 250-350, 250-400, 350-400, 300-400, or 250-400 mg of tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil, and 200 mg of emtricitabine. In some embodiments, the antibodies or antigen-binding fragments described herein are combined with 300 mg of tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil, and 200 mg of emtricitabine. The antibody or antigen-binding fragment can be combined with the agent provided herein at any dosage (e.g., 1 mg to 500 mg of the antibody or antigen-binding fragment) in the same manner that each combination of dosages is specifically and individually recited.

[0285] Long-acting HIV inhibitor In some embodiments, the antibodies or antigen-binding fragments described herein can be co-administered with a long-acting HIV inhibitor. Examples of agents developed as long-acting HIV inhibitors include, but are not limited to: cabotegravir LA, rilpivirine LA, any integrase LA, VM-1500 LAI, maraviroc (LAI), tenofovir implant, MK-8591 implant, long-acting dolutegravir.

[0286] In one embodiment, the kit comprises an antibody or antigen-binding fragment described herein combined with one or more (e.g., 1, 2, 3, 1 or 2, or 1-3) additional therapeutic agents.

[0287] HIV vaccine In some embodiments, the antibodies or antigen-binding fragments described herein are combined with an HIV vaccine. Examples of HIV vaccines include peptide vaccines, recombinant subunit protein vaccines, live vector vaccines, DNA vaccines, HIV MAG DNA vaccines, CD4-derived peptide vaccines, combinations of vaccines, adenovirus vector vaccines (e.g., Ad5, Ad26, or Ad35), simian adenoviruses (chimpanzee, gorilla, rhesus, i.e., rhAd), adeno-associated virus vector vaccines, chimpanzee adenovirus vaccines (e.g., ChAdOX1, ChAd68, ChAd3, ChAd63, ChAd83, ChAd155, ChAd157, Pan5, Pan6, Pan7, Pan9), coxsackievirus-based vaccines, enterovirus-based vaccines, gorilla adenovirus vaccines, lentivirus vector-based vaccines, two-segment or three-segment arenavirus-based vaccines (e.g., LCMV, Pichinde), trimer-based HIV-1 vaccines, measles virus-based vaccines, flavivirus vector-based vaccines, tobacco mosaic virus vector-based vaccines, varicella zoster virus-based vaccines, human parainfluenza virus 3 (PIV3)-based vaccines, poxvirus-based vaccines (modified vaccinia virus Ankara (MVA), NYVAC derived from orthopoxvirus, and ALVAC (canarypox virus) strain derived from avipoxvirus); fowlpox virus-based vaccines, rhabdovirus-based vaccines such as vesicular stomatitis virus (VSV) and Maraba virus; recombinant human CMV (rhCMV)-based vaccines, alphavirus-based vaccines such as Semliki Forest virus, Venezuelan equine encephalitis virus, Sindbis virus (see, e.g., Lauer, et al., Clin Vaccine Immunol. (2017) 24(1): e00298-16); LNP-formulated mRNA-based therapeutic vaccines; and self-replicating RNA / self-amplifying RNA vaccines formulated with LNP.

[0288] Examples of HIV vaccines include anti-CD40.Env-gp140 vaccine, Ad4-EnvC150, BG505 SOSIP.664 gp140 adjuvant vaccine, BG505 SOSIP.GT1.1 gp140 adjuvant vaccine, Chimigen HIV vaccine, ConM SOSIP.v7 gp140, rgp120 (AIDSVAX), ALVAC HIV (vCP1521 / AIDSVAX B / E (gp120) (RV144), monomeric gp120 HIV-1 subtype C vaccine, MPER-656 liposome subunit vaccine, Remune, ITV-1, Contre Vir, Ad5-ENVA-48, DCVax-001 (CDX-2401), Vacc-4x, Vacc-C5, VAC-3S, multiclade DNA recombinant adenovirus-5 (rAd5), rAd5 gag-pol env A / B / C vaccine, Pennvax-G, Pennvax-GP, Pennvax-G / MVA-CMDR, HIV-TriMix-mRNA vaccine, HIV-LAMP-vax, Ad35, Ad35-GRIN, NAcGM3 / VSSP ISA-51, poly-ICLC adjuvant vaccine, TatImmune, GTU-multiHIV (FIT-06), ChAdV63.HIVconsv, gp140δV2.TV1+ MF-59, rVSVIN HIV-1 gag vaccine, SeV-EnvF, SeV-Gag vaccine, AT-20, DNK-4, ad35-Grin / ENV, TBC-M4, HIVAX, HIVAX-2, N123-VRC-34.01 Inducible epitope-based HIV vaccines, NYVAC-HIV-PT1, NYVAC-HIV-PT4, DNA-HIV-PT123, rAAV1-PG9DP, GOVX-B11, GOVX-B21, GOVX-C55, TVI-HIV-1, Ad-4 (Ad4-env Clade C+Ad4-mGag), Paxvax, EN41-UGR7C, EN41-FPA2, ENOB-HV-11, PreVaxTat, AE-H, MYM-V101, CombiHIVvac, ADVAX, MYM-V201, MVA-CMDR, MagaVax, DNA-Ad5 gag / pol / nef / nev (HVTN505), MVATG-17401, ETV-01, CDX-1401, DNA and Sev vector vaccines expressing SCaVII, rcAD26.MOS1.HIV-Env, Ad26.Mod.HIV vaccine, Ad26.Mod.HIV+MVA mosaic vaccine+gp140, AGS-004, AVX-101, AVX-201, PEP-6409, SAV-001, ThV-01, TL-01, TUTI-16, VGX-3300, VIR-1111, IHV-001, and virus-like particle vaccines such as pseudovirion vaccines, CombiVICHvac, LFn-p24 B / C fusion vaccine, GTU-based DNA vaccine, HIV gag / pol / nef / env DNA vaccine, anti-TAT HIV vaccine, conjugate polypeptide vaccine, dendritic cell vaccine, gag-based DNA vaccine, GI-2010, gp41 HIV-1 vaccine, HIV vaccine (PIKA adjuvant), I i-key / MHC class II epitope hybrid peptide vaccine, ITV-2, ITV-3, ITV-4, LIPO-5, multi-clade Env vaccine, MVA vaccine, Pennvax-GP, pp71-deficient HCMV vector HIV gag vaccine, recombinant peptide vaccine (HIV infection), NCI, rgp160 HIV vaccine, RNActive HIV vaccine, SCB-703, Tat Oyi vaccine, TBC-M4, therapeutic HIV vaccine, UBI HIV gp120, Vacc-4x+romidepsin, mutant gp120 polypeptide vaccine, rAd5 gag-pol env A / B / C vaccine, DNA.HTI and MVA.HTI, VRC-HIVDNA016-00-VP + VRC-HIVADV014-00-VP, INO-6145, JNJ-9220, gp145 C.6980; eOD-GT8 60mer-based vaccine, PD-201401, env(A, B, C, A / E) / gag(C) DNA vaccine, gp120(A, B, C, A / E) protein vaccine, PDPHV-201401, Ad4-EnvCN54, EnvSeq-1 Envs HIV-1 vaccine (with GLA-SE adjuvant), HIV p24gag prime-boost plasmid DNA vaccine, HIV-1 iglb12 neutralizing VRC-01 antibody-stimulating anti-CD4 vaccine, MVA-BN HIV-1 vaccine therapy, UBI HIV gp120, mRNA-based prophylactic vaccine, VPI-211, and TBL-1203HI, among others, but not limited to these.

[0289] D. Kit In other aspects, the disclosure further provides kits useful for performing diagnostic and prognostic assays using the antibodies, polypeptides, and nucleic acids of the invention. The kits of the invention include a suitable container containing an HIV antibody, polypeptide, or nucleic acid of the invention in either labeled or unlabeled form. Further, when the antibody, polypeptide, or nucleic acid is provided in a labeled form suitable for an indirect binding assay, the kit further includes reagents for performing a suitable indirect assay. For example, the kit may include one or more suitable containers containing an enzyme substrate or derivatizing agent, depending on the nature of the label. It may also include control samples and / or instructions for use. The invention also provides kits for detecting the presence of an HIV antibody of the invention or the nucleotide sequence of an HIV antibody in a biological sample by PCR or mass spectrometry.

[0290] In some embodiments, the kit further comprises a pharmaceutically acceptable dosage unit of a pharmaceutically effective amount of an anti-HIV agent. The two pharmaceutically acceptable dosage units may be in the form of a single pharmaceutically acceptable dosage unit. In some embodiments, the anti-HIV agent is selected from the group consisting of non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry inhibitors or fusion inhibitors, and integrase inhibitors.

[0291] In some embodiments, the kit further comprises a container containing the composition and optionally may contain informational materials. The informational materials can be explanatory, educational, commercial, or other materials related to the methods described herein and / or the use of the agent for therapeutic benefit. In one embodiment, the kit further comprises an additional therapeutic agent as described above. For example, the kit comprises a first container containing the composition and a second container for the additional therapeutic agent.

[0292] The informational materials of the kit are not limited in their form. In some embodiments, the informational materials can include information regarding the manufacture, concentration, expiration date, batch, or manufacturing site of the composition. In one embodiment, the informational materials relate to a method of treating a subject in need thereof by administering the composition, for example, in a suitable dosage, dosage form, or mode of administration (e.g., the dosage, dosage form, or mode of administration described herein). In one embodiment, the instructions for use provide a dosing plan, dosing schedule, and / or route of administration of the composition or additional therapeutic agent. The information can be provided in various forms, including printed text, computer-readable materials, video recordings, audio recordings, or information including links or addresses to substantial materials.

[0293] The kit may include one or more containers for the composition. In some embodiments, the kit includes separate containers, partitions, or compartments for the composition and the information material. For example, the composition may be placed in a bottle or vial, and the information material may be placed in a plastic sleeve or packet. In other embodiments, the individual elements of the kit are included in a single undivided container. For example, the composition is included in a bottle or vial with the information material attached thereto in the form of a label. In some embodiments, the kit includes a plurality (e.g., pack) of individual containers, each containing one or more unit dosage forms of the agent (e.g., the dosage forms described herein).

[0294] The kit may include an apparatus suitable for administration of the composition or other suitable delivery device. The apparatus may be pre-loaded and provided with one or both of the agents, or may be emptied but is suitable for loading. Such a kit may include a syringe capable of injecting an antibody contained in the kit into an animal such as a human.

[0295] E. Definitions To assist in understanding the detailed description of the compositions and methods according to the present disclosure and to facilitate the clear disclosure of various aspects of the disclosure, some clear definitions are provided. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0296] As used herein, the term "antibody" (Ab) includes monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies and polyreactive antibodies), and antibody fragments. Thus, the term "antibody" as used in any context herein means any specific binding member, immunoglobulin class and / or isotype (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA, IgD, IgE, and IgM); and includes, but is not limited to, biologically relevant fragments or specific binding members thereof including Fab, F(ab’)2, Fv, and scFv (single-chain or related entities). In the art, an antibody is understood to be a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds or its antigen-binding portion. The heavy chain is composed of a heavy-chain variable region (VH) and heavy-chain constant regions (CH1, CH2, and CH3). The light chain is composed of a light-chain variable region (VL) and light-chain constant region (CL). The variable regions of both the heavy and light chains are composed of framework regions (FWR) and complementarity-determining regions (CDR). The four FWR regions are relatively conserved, while the CDR regions (CDR1, CDR2, and CDR3) represent hypervariable regions and are arranged as follows from the NH2 terminus to the COOH terminus: FWR1, CDR1, FWR2, CDR2, FWR3, CDR3, FWR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens, while, depending on the isotype, the constant regions may mediate binding of the immunoglobulin to host tissues or factors.

[0297] The definition of "antibody" as used herein also includes chimeric antibodies, humanized antibodies, and recombinant antibodies, transgenic non-human animals, and human antibodies produced from antibodies selected from libraries using enrichment techniques available to those skilled in the art.

[0298] "Neutralizing antibody" neutralizes the ability of HIV to initiate and / or persist in a host and / or target cells in vitro. The present disclosure provides neutralizing human monoclonal antibodies, which antibodies recognize an antigen from HIV, such as the gp120 polypeptide. In some embodiments, a "neutralizing antibody" has a neutralization index > 1.5 or > 2.0 and can inhibit the entry of HIV-1 viruses, such as SF162 and / or JR-CSF (Kostrikis LG et al., J. Virol., 70(1): 445-458 (1996)).

[0299] In some embodiments, the neutralizing antibody is a broadly neutralizing antibody (such as a monoclonal antibody) that targets HIV-L. The term "broadly neutralizing antibody" refers to an antibody that neutralizes multiple HIV-1 virus species (from diverse clades and different strains within a clade) in a neutralization assay. A broadly neutralizing antibody can neutralize at least 2, 3, 4, 5, 6, 7, 8, 9 or more different HIV-1 strains, strains belonging to the same or different clades. In some embodiments, a broadly neutralizing antibody can neutralize multiple HIV-1 species belonging to at least 2, 3, 4, 5, or 6 different clades. In some embodiments, the inhibitory concentration of the antibody is less than about 0.0001 pg / mL, less than about 0.001 pg / mL, less than about 0.01 pg / mL, less than about 0.1 pg / mL, less than about 0.5 pg / mL, less than about 1.0 pg / mL, less than about 5 pg / mL, less than about 10 pg / mL, less than about 25 mg / mL, less than about 50 mg / mL, or less than about 100 mg / mL to neutralize about 50% of the input virus in a neutralization assay.

[0300] "Antibody fragment" includes a part of an intact antibody such as the antigen-binding or variable region of an intact antibody. Examples of antibody fragments include Fab, Fab’, F(ab’)2, and Fv fragments; diabodies; linear antibodies (see, e.g., U.S. Patent No. 5,641,870; Zapata et al., Protein Eng. 8(10): 1057-1062

[1995] ); single-chain antibody molecules; single-chain variable fragments; and multispecific antibodies formed from antibody fragments, but are not limited thereto.

[0301] The term "variable" refers to the fact that certain segments of the variable (V) domains vary extensively in sequence among antibodies. The V domains mediate antigen binding and define the specificity of a particular antibody for a particular antigen. However, the variability is not evenly distributed throughout the entire 110-amino acid span of the variable regions. Instead, the V regions are composed of relatively invariant stretches called framework regions (FRs) of 15-30 amino acids, separated by short regions of extreme variability called "hypervariable regions" that are 9-12 amino acids in length. The native variable regions of the heavy and light chains each consist of four FRs, adopt mainly a beta-sheet conformation, are connected by three hypervariable regions, form loops, and in some cases form part of the beta-sheet structure. The hypervariable regions of each chain are held in close proximity by the FRs and, together with the hypervariable regions of the other chain, contribute to the formation of the antigen-binding site of the antibody (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).

[0302] As used herein, the term "hypervariable region" refers to the amino acid residues of an antibody that are involved in antigen binding. The hypervariable regions typically include amino acid residues from "complementary determining regions" ("CDRs").

[0303] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies that make up the population are identical except for naturally occurring mutations that may be present in minor amounts. The term "polyclonal antibody" refers to a preparation that contains different antibodies directed against different determinants ("epitopes").

[0304] As used herein, "subject" refers to humans and non-human animals. Examples of non-human animals include all vertebrates, such as non-human mammals, non-human primates (especially higher primates), dogs, rodents (e.g., mice or rats), guinea pigs, cats, and rabbits, and other mammals, as well as non-mammals such as birds, amphibians, and reptiles. In one embodiment, the subject is a human. In other embodiments, the subject is an experimental animal or an animal suitable as a disease model.

[0305] The term "polypeptide" is used in its conventional sense, i.e., as an amino acid sequence. The polypeptide is not limited to a particular length of the product. Peptides, oligopeptides, and proteins are included in the definition of polypeptide, and such terms may be used interchangeably herein unless otherwise specified. This term also includes post-expression modifications of polypeptides, such as glycosylation, acetylation, phosphorylation, etc., as well as other modifications known in the art, both naturally occurring and non-naturally occurring. A polypeptide can be the entire protein or a subsequence thereof. The particular polypeptides of interest in the context of the present invention are amino acid subsequences containing CDR, VH, and VL that can bind to an antigen or an HIV-infected cell.

[0306] As used herein, the polypeptide "variant" is a polypeptide that is usually different from the polypeptides specifically disclosed herein in one or more substitutions, deletions, additions and / or insertions. Such variants can occur naturally or can be produced synthetically, for example, by modifying one or more of the above polypeptide sequences of the invention using any of several techniques described herein and / or known in the art, and evaluating one or more biological activities of the polypeptide.

[0307] "Homology" or "sequence identity" refers to the percentage of residues of a polynucleotide or polypeptide sequence variant that is identical to the non-variant sequence, after aligning the sequences and introducing gaps as necessary to achieve the maximum percentage of homology. In certain embodiments, the polynucleotide and polypeptide variants have at least about 70%, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least about 98%, or at least about 99% polynucleotide or polypeptide homology to the polynucleotides or polypeptides described herein.

[0308] Such variant polypeptide sequences share at least 70% (i.e., 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity with the sequences described in this application. In further embodiments, the described invention provides polypeptide fragments comprising contiguous stretches of various lengths of the amino acid sequences disclosed herein. For example, the present invention provides peptide sequences comprising peptides of at least about 5, 10, 15, 20, 30, 40, 50, 75, 100, 150, or more contiguous, and all intermediate lengths therebetween, of one or more of the sequences disclosed herein.

[0309] The terms "nucleic acid" and "polynucleotide" are used interchangeably herein and refer to single-stranded or double-stranded RNA, DNA, or hybrid polymers. Polynucleotides can include genomic sequences, extra-genomic and plasmid sequences, and smaller engineered genetic segments that express or are adapted to express polypeptides.

[0310] An "isolated nucleic acid" is a nucleic acid substantially separated from other genomic DNA sequences, as well as proteins or complexes such as ribosomes and polymerases that naturally accompany the native sequence. The term includes nucleic acid sequences removed from their natural environment, including recombinant or cloned DNA isolates, and analogs synthesized biologically by a heterologous system or chemically synthesized analogs. Substantially pure nucleic acids include nucleic acids in isolated form. Thus, this refers to the initially isolated nucleic acid and does not exclude genes or sequences subsequently added by human hand to the isolated nucleic acid.

[0311] As used herein, a polynucleotide "variant" is a polynucleotide that typically differs from a polynucleotide specifically disclosed herein in one or more substitutions, deletions, additions, and / or insertions. Such variants can occur naturally or can be produced synthetically, for example, by modifying one or more of the polynucleotide sequences of the invention using any of several techniques described herein and / or known in the art, and evaluating one or more of the biological activities of the encoded polypeptides.

[0312] As used herein, the term "cell" can be any cell, including but not limited to cells of eukaryotic multicellular species such as mammalian or human cells (e.g., as contrasted with single-celled yeast cells). A cell can exist as a single entity or as part of a larger population of cells. Such "larger populations of cells" can include, for example, cell cultures (either mixed or pure), tissues (e.g., endothelial, epithelial, mucosal or other tissues), organs (e.g., lung, liver, muscle and other organs), organ systems (e.g., cardiovascular, respiratory, gastrointestinal, urinary, nervous, integumentary or other organ systems), or organisms (e.g., birds, mammals, etc.).

[0313] As used herein, "label" refers to a detectable compound or composition that is directly or indirectly conjugated to an antibody to produce a "labeled" antibody. A label can also be conjugated to the polypeptides and / or nucleic acid sequences disclosed herein. The label can be detectable by itself (e.g., a radioisotope label or a fluorescent label), or in the case of an enzyme label, can catalyze a chemical change in a detectable substrate compound or composition. The described antibodies and polypeptides of the invention can also be modified to include, for example, an epitope tag or a label for use in purification or diagnostic applications. Suitable means of detection include, but are not limited to, the use of labels such as radiolabeled nucleotides, enzymes, coenzymes, fluorophores, chemiluminescent agents, chromogens, enzyme substrates or cofactors, enzyme inhibitors, cofactor complexes, free radicals, particles, dyes, etc.

[0314] The term "assessing" includes all forms of measurement and also includes determining whether an element is present. The terms "determining", "measuring", "assessing", "calculating", and "analyzing" are used interchangeably and include both quantitative and qualitative determinations. An assessment can be relative or absolute. "Assessment of existence" includes determining the amount of what is present and / or determining whether it is present or absent. As used herein, the terms "determining", "measuring", "assessing", and "analyzing" are used interchangeably and include both quantitative and qualitative determinations.

[0315] The terms "treating" or "treatment" or "alleviating" are used interchangeably to refer to both therapeutic treatment and prophylactic or preventative measures; the purpose is to prevent or slow (reduce) the targeted pathologic condition or disorder. Persons in need of treatment include not only those already having the disorder but also those prone to having the disorder or those in whom the disorder is to be prevented. A subject or mammal is successfully "treated" for an infectious disease if, after administration of a therapeutically effective amount of an antibody according to the methods of the invention, the patient exhibits an observable and / or measurable decrease, regardless of the presence or absence of one or more of the following: a decrease in the number of infected cells or the absence of infected cells; a decrease in the proportion of total cells that are infected; and / or alleviation to some extent of one or more of the symptoms associated with a particular infectious disease; a decrease in morbidity and mortality, as well as an improvement in quality of life issues. The above parameters for assessing the success and improvement of treatment of a disease are readily measurable by routine procedures well known to physicians.

[0316] "Effective amount" or "therapeutically effective amount" refers to the amount of a compound or agent that can produce a medically desirable result in a subject being treated. The treatment can be carried out in vivo or ex vivo, alone or in combination with other drugs or therapies. A therapeutically effective amount can be administered in one or more administrations, applications, or dosages and is not intended to be limited to a particular formulation or route of administration.

[0317] As used herein, the term "disease" is generally intended to be synonymous with the terms "disorder" and "condition" (such as in the case of a medical condition), and is used interchangeably to reflect any abnormal condition of the human or animal body, or a part thereof that impairs normal function, which typically manifests by distinguishing signs and symptoms and causes a decrease in the duration or quality of life of a human or animal.

[0318] All of the terms "reduce", "decrease", "decrement", "reduction", or "inhibit" are used herein generally to mean a statistically significant amount of decrease. However, for clarity or to avoid misunderstanding, "reduced", "decreased" or "reduced" or "inhibited" means a decrease of at least 10% compared to a reference level, for example, at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% of the decrease compared to the reference level, or a decrease up to 100% (e.g., no value compared to a reference sample), or a decrease of 10 - 100%.

[0319] As used herein, the term "modulate" means to refer to any change in a biological state, i.e., an increase, a decrease, etc.

[0320] The terms "increased", "increasing", "enhanced", or "activated" are all used herein to normally mean an increase in a statically significant amount; to avoid misunderstanding, the terms "increased", "increasing", "enhanced", or "activated" mean an increase of at least 10% compared to a reference level, for example, at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or up to 100% increase compared to the reference level, or an increase between 10 - 100%, or an increase of at least about 2-fold, or at least about 3-fold, or at least about 4-fold, or at least about 5-fold, or at least about 10-fold, or any increase between 2 - 10-fold compared to the reference level.

[0321] The terms "effective amount", "effective dosage", or "effective dose" are defined as an amount sufficient to achieve, or at least partially achieve, a desired effect. The "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of the drug that, when used alone or in combination with other therapeutic agents, promotes the regression of a demonstrated disease by reducing the severity of the disease symptoms, increasing the frequency and duration of periods without disease symptoms, or preventing the disability or damage caused by the pain of the disease. The "preventively effective amount" or "preventively effective dose" of a drug is the amount of the drug that, when administered to a subject at risk of developing a disease or suffering a recurrence of the disease, alone or in combination with other therapeutic agents, suppresses the onset or recurrence of the disease. The ability of a therapeutic or preventive agent to promote the regression of a disease or suppress the onset or recurrence of a disease can be evaluated using various methods known to those of skill in the art, such as in human subjects during clinical trials, animal model systems that predict efficacy in humans, or assays of the activity of the agent in in vitro assays.

[0322] Thus, a dosage expressed as [g, mg, or other unit] / kg (or g, mg, etc.) typically refers to [g, mg, or other unit] "per 1 kg (or g, mg, etc.) of body weight", even when the term "body weight" is not explicitly mentioned.

[0323] As used herein, the term "agent" is used to denote a compound, a mixture of compounds, a biological macromolecule (e.g., a nucleic acid, an antibody, a protein or a part thereof, e.g., a peptide), or an extract made from biological materials such as cells or tissues of bacteria, plants, fungi, or animals (especially mammals). The activity of such an agent may render it suitable as a "therapeutic agent", which is a biologically, physiologically, or pharmacologically active substance (or substances) that acts locally or systemically in a subject.

[0324] The terms "therapeutic agent", "therapeutically capable agent", or "medicament" are used interchangeably to refer to a molecule or compound that confers some beneficial effect upon administration to a subject. Such beneficial effects include enabling a diagnostic determination; improving a disease, symptom, disorder, or pathological condition; reducing or preventing the onset of a disease, symptom, disorder, or condition; and generally counteracting a disease, symptom, disorder, or pathological condition.

[0325] As used herein, "combination" therapy means, unless otherwise clearly indicated from the context, the administration of two or more therapeutic agents in a coordinated manner, including, but not limited to, co - administration. Specifically, combination therapy includes both co - administration (e.g., administration of co - formulated or separate therapeutic compositions simultaneously) and sequential or continuous administration under the condition that the administration of one therapeutic agent is conditioned in some way on the administration of another therapeutic agent. For example, one therapeutic agent may be administered only after another therapeutic agent has been administered and allowed to act for a predetermined period. See, e.g., Kohrt et al. (2011) Blood 117:2423. Administration "in combination with" one or more additional therapeutic agents includes simultaneous (concurrent) and sequential administration in either order.

[0326] The terms "sample", "test sample", and "patient sample" may be used interchangeably herein. The sample can be a sample of serum, urine plasma, amniotic fluid, cerebrospinal fluid, cells (e.g., antibody-producing cells) or tissue. Such a sample can be used directly as obtained from a patient as discussed herein or as known in the art, or can be pretreated by filtration, distillation, extraction, concentration, centrifugation, inactivation of interfering components, addition of reagents, etc. in order to modify the properties of the sample in some way. The terms "sample" and "biological sample" as used herein generally refer to biological material that is being tested for and / or suspected of containing an analyte of interest such as an antibody. The sample can be any tissue sample from the subject. The sample can contain proteins from the subject.

[0327] As used herein, the terms "inhibit" and "antagonize" mean to cause a measurable decrease or to completely prevent the expression, stability, function or activity of a molecule, reaction, interaction, gene, mRNA, and / or protein. An inhibitor is, for example, a compound that binds to a stimulus, partially or completely blocks it, or decreases, prevents, delays, inactivates, desensitizes, or down-regulates the stability, expression, function, and activity of proteins, genes, and mRNAs, such as an antagonist.

[0328] Examples of "parenteral" administration of a composition include subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, or infusion techniques.

[0329] As used herein, the term "pharmaceutical composition" refers to a mixture of at least one compound useful within the present invention and other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. The pharmaceutical composition facilitates the administration of the compound to a living being.

[0330] A number of techniques for administering compounds exist in the art, including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, and topical administration.

[0331] As used herein, the term "pharmaceutically acceptable" refers to a material, such as a carrier or diluent, that does not abrogate the biological activity or properties of a composition and is relatively non-toxic, i.e., the material can be administered to an individual without causing undesirable biological effects or interacting in a harmful manner with any of the components of the composition in which it is contained.

[0332] The term "pharmaceutically acceptable carrier" includes pharmaceutically acceptable salts, pharmaceutically acceptable materials, compositions, or carriers such as liquid or solid fillers, diluents, excipients, solvents or encapsulating materials, which are involved in transporting the compounds of the present invention within or to a subject and enable the intended functions to be performed. Typically, such compounds are carried or transported from one organ or part of the body to another. Each salt or carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the subject. Some examples of materials that can be useful as pharmaceutically acceptable carriers include the following: sugars such as lactose, glucose, sucrose; starches such as corn starch and potato starch; cellulose and cellulose derivatives such as carboxymethylcellulose sodium, ethyl cellulose, cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solution; diluents; granulating agents; lubricants; binders; disintegrants; wetting agents; emulsifiers; coloring agents; release agents; coating agents; sweetening agents; flavoring agents; fragrances; preservatives; antioxidants; plasticizers; gelling agents; thickening agents; hardening agents; setting agents; suspending agents; surfactants; humectants; carriers; stabilizers; and other non-toxic compatible substances used in the formulation, or any combination thereof. As used herein, "pharmaceutically acceptable carrier" also includes all coatings, antibacterial and antifungal agents, and absorption delaying agents, etc., that are compatible with the activity of the compound and physiologically acceptable to the subject. Supplementary active compounds can also be incorporated into the composition.

[0333] As used herein, the term "pharmaceutically acceptable salt" refers to salts of an administered compound prepared from pharmaceutically acceptable non-toxic acids such as inorganic acids, organic acids, solvates, hydrates, or their clathrates.

[0334] As used herein, the term "in vitro" refers to events that occur in an artificial environment, such as in a test tube or reaction vessel, cell culture, etc., rather than within a multicellular organism.

[0335] As used herein, the term "in vivo" refers to events that occur within a multicellular organism such as a non-human animal.

[0336] It should be noted that as used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural references unless the context clearly dictates otherwise.

[0337] The terms "listed", "comprising", "including", or "having" and their variations mean including the items recited thereafter and their equivalents, as well as additional items, unless otherwise specified.

[0338] Phrases such as "in one embodiment", "in various embodiments", "in some embodiments", etc. are used repeatedly. Such phrases do not necessarily refer to the same embodiment, but they may do so unless the context indicates otherwise.

[0339] The term "and / or" or " / " means any one of the items, any combination of the items, or all of the items associated with this term.

[0340] The word "substantially" does not exclude "completely". For example, a composition that "substantially does not contain" Y may completely not contain Y. If necessary, the word "substantially" may be omitted from the definition of the present invention.

[0341] As used herein, the terms “about” or “approximately” when applied to one or more values of interest, refer to a value similar to the recited reference value. In some embodiments, the term “about” or “approximately” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, in either direction (greater than or less than) of the recited reference value, unless otherwise specified or otherwise apparent from the context (except where such numbers would exceed 100% of the possible value). Unless otherwise stated herein, the term “about” is intended to include values approaching the recited range that are equivalent with respect to the functionality of the individual components, the composition, or the embodiment, e.g., including weight percentages.

[0342] It should be understood that when values and ranges are provided herein, all values and ranges included within these values and ranges are meant to be included within the scope of the present invention. Further, all values within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

[0343] As used herein, the term “each” when used in reference to a collection of items, is intended to identify individual items within the collection, but does not necessarily refer to all items within the collection. Exceptions can occur where an explicit disclosure or context clearly indicates otherwise.

[0344] Any examples, or use of exemplary language (e.g., “such as”) provided herein are merely intended to better illuminate the invention and do not limit the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0345] All methods described in this specification are performed in any suitable order, unless otherwise indicated herein or unless clearly inconsistent with the context. For any of the provided methods, the steps of the method may occur simultaneously or sequentially. If the steps of the method occur sequentially, unless otherwise specified, the steps may occur in any order.

[0346] If a method includes a combination of steps, unless otherwise described herein, all combinations or sub - combinations of the steps are included within the scope of this disclosure.

[0347] Each publication, patent application, patent, and other reference cited herein is hereby incorporated by reference in its entirety to the extent not inconsistent with this disclosure. The publications disclosed herein are provided only for their disclosure prior to the filing date of the present invention. Nothing in this specification should be construed as an admission that the present invention is not entitled to antedate such publications by virtue of prior invention. Further, the provided publication dates may be different from the actual publication dates and may need to be individually verified.

[0348] It is understood that the examples and embodiments described in this specification are for illustrative purposes only, and that various modifications or changes from that perspective are proposed to those skilled in the art and should be included within the spirit and scope of this application and the appended claims.

[0349] F. Examples Example 1 In this example, the materials and methods used in the following subsequent examples are described.

[0350] Multireactivity assay: As described (Hotzel et al. 2012, mAbs, 4:6, 753-760), the multireactivity assay was performed using ELISA detection of nonspecific binding to baculovirus extracts. Briefly, a solution of 1% baculovirus particles in 100 mM sodium bicarbonate buffer pH 9.6 was absorbed into the wells of a 384-well ELISA plate (Nunc Maxisorp) using a Tecan Freedom Evo liquid handling robot, and the plate was incubated overnight at 4 °C. The plate was blocked with 0.5% BSA in PBS for 1 h at room temperature. Purified IgG (diluted to 1 μg / mL in PBS, 0.5% BSA) was added to the blocked assay plate and incubated for 3 h at room temperature. Bound IgG was detected as a luminescence signal at 425 nm using an HRP-conjugated anti-human IgG (H&L) secondary antibody (Genscript) and SuperSignal ELISA Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific).

[0351] Neutralization assay: HIV neutralizing potency was determined using a luciferase-based TZM-bl assay as described above (Montefiori, D. C. 1019, Methods Mol. Biol. 485, 395-405).

[0352] Half-life in non-human primates: The half-life in cynomolgus monkeys was determined as described (Shingai, M. et al., 2014, J. Exp. Med. 211, 2061-74). Briefly, cynomolgus monkeys were administered 10 mg / kg of purified IgG, and antibodies in serum were detected over 70 days by ELISA and / or neutralizing activity. The half-life was determined by fitting the data from days 8-70.

[0353] Mouse Pharmacokinetics: The designed reagent was radiolabeled with 125I using the iodination method and purified by SEC (Yazaki PJ et al., 2008, Nucl Med Biol., 35(2), 151-8). The half-life and tissue distribution in mice were determined as described above (Yazaki PJ et al., 2013, PEDS 26(3), 187-193). Briefly, NSG mice were injected with mouse IVIG (4 hours prior), and then approximately 3 μg of labeled Ab was injected by tail vein injection (approximately 3-6 microcuries). Blood samples were collected and the radiolabeled protein was counted. These data were plotted as a function of time of blood clearance. Mice were sacrificed at 96 hours, and the weights of organs and carcasses were weighed, counted, and the numbers were normalized to body weight and plotted.

[0354] Example 2 The parental antibody NIH45-46G54W consists of NIH45-46 HC and NIH45-46HC with the G54W mutation paired with NIH45-46LC. The novel antibody variants have mutations in one or both of these chains.

[0355] The single chain from which the novel antibody variant is constructed is as follows:

[0356] [Table 4]

[0357] The novel antibody reagent consists of the following HC / LC pairs (LS indicates the mutations described above in the Fc region, e.g., M428L / N434S): HC1 / SAP10 HC1 / SAP10-LS HC1 / SAP10T HC1 / SAP10T-LS HC1 / SAP8 HC1 / SAP3 HC1 / NIH45-46 LC HC1 / m2 NIH45-46 G54W / SAP10 NIH45-46 G54W / SAP3 NIH45-46 G54W / SAP8 The in vivo half-life of G54W mutants of NIH45-46 (such as NIH45-46G54W and NIH45-46m2) is short (Shingai, M. et al., 2014, J. Exp. Med. 211, 2061-74). The short half-life correlates with high levels of polyreactivity (Hotzel, I., et al., 2012, mAbs 4, 753-760). Our newly designed mutant HC1 / SAP10 shows reduced polyreactivity (Figure 2) and retains potency (Figure 1, Tables 1 and 2). The neutralizing power of HC1 / SAP10 is slightly reduced compared to NIH45-46G54W and NIH45-46m2, but still comparable to other anti-HIV bNAbs such as N6 and 3BNC117 (Figure 1).

[0358] The half-life of bNAb NIH45-46 m2 in non-human primates (NHPs) could not be determined due to rapid clearance (Shingai, M. et al., 2014, J. Exp. Med. 211, 2061-74). In contrast, our new mutant HC1 / SAP10-LS has a half-life and tissue distribution similar to that of mouse 3BNC117 (Figure 3), and the half-life in NHPs is 8.4 days.

[0359] The neutralization data of the new reagents are shown in Table 1 and summarized in Table 2.

[0360]

Table 5-1

[0361]

Table 5-2

[0362]

Table 5-3

[0363]

Table 6-1

[0364]

Table 6-2

[0365]

Table 7

Claims

**Claim 1** An isolated anti-HIV antibody, or an antigen-binding portion thereof, comprising a heavy chain having a heavy chain amino acid sequence that is at least 95% identical to SEQ ID NO: 1 and a light chain having a light chain amino acid sequence that is at least 95% identical to SEQ ID NO: 2, wherein the heavy chain complementarity-determining regions (CDRs) H1, H2, and H3 of the anti-HIV antibody, or the antigen-binding portion thereof, are the same as CDRs H1, H2, and H3 of SEQ ID NO: 1, the light chain complementarity-determining regions (CDRs) L1, L2, and L3 of the anti-HIV antibody, or the antigen-binding portion thereof, are the same as CDRs L1, L2, and L3 of SEQ ID NO: 2, wherein (i) the heavy chain amino acid sequence comprises one or more substitutions at one or more residues selected from the group consisting of S5, S21, G54, G55, V57, T68, T70, V73, S75, F79, S82, D85, V89, and P112; or (ii) the light chain amino acid sequence comprises one or more substitutions at one or more residues selected from the group consisting of T5, S7, T10, S12, S14, T18, I20, S22, S61, S63, W65, D68, N70, S72, S74, and S78, the numbering of the residues being shown by Kabat numbering, an isolated anti-HIV antibody, or an antigen-binding portion thereof. **Claim 2** An isolated anti-HIV antibody, or an antigen-binding portion thereof, comprising a heavy chain having heavy chain complementarity-determining regions (CDRs) H1, H2, and H3 and a light chain having light chain complementarity-determining regions (CDRs) L1, L2, and L3, wherein the CDRs H1, H2, and H3 are the same as CDRs H1, H2, and H3 of SEQ ID NO: 1 (except for a heavy chain having a mutation at residue Y97), and the CDRs L1, L2, and L3 are the same as CDRs L1, L2, and L3 of SEQ ID NO: 2 (except for a light chain having a mutation at one or more residues selected from the group consisting of R24, Q27, S28, S30, and R59), the numbering of the residues being shown by Kabat numbering, an isolated anti-HIV antibody, or an antigen-binding portion thereof. **Claim 3** The isolated anti-HIV antibody or antigen-binding portion thereof according to claim 1 or 2, wherein the heavy chain amino acid sequence comprises one or more of the following substitutions: Substitution of S5D or S5E; Substitution of S21D or S21E; Substitution of G54W or conservative substitution of Trp at residue G54; Substitution of G55D or G55E; Substitution of V57D or V57E; Substitution of T68D or T68E; Substitution of T70D or T70E; Substitution of V73D or V73E; Substitution of S75D or S75E; Substitution of F79D, F79E, F79Y or F79H, or conservative substitution of Tyr or His at residue F79; Substitution of S82D or S82E; Substitution of D85E or conservative substitution of Asp at residue D85; Substitution of V89D or V89E; Substitution of P112D or P112E; Or a combination thereof.

4. The isolated anti-HIV antibody or antigen-binding portion thereof according to any one of claims 1 to 3, wherein the light chain amino acid sequence comprises one or more of the following substitutions: Substitution of T5D or T5E; Substitution of S7D or S7E; Substitution of T10D or T10E; Substitution of S12D or S12E; Substitution of S14D or S14E; Substitution of T18D or T18E; Substitution of I20D or I20E; Substitution of S22D or S22E; Substitution of R59D or R59E; Substitution of S61D or S61E; Substitution of S63D or S63E; Substitution of W65D, W65E, W65Y or W65H, or conservative substitution of Tyr or His at residue W65; Substitution of D68E or conservative substitution of Asp at residue D68; Substitution of N70D or N70E; Substitution of S72D or S72E; Substitution of S74D or S74E; Substitution of S78D or S78E; Or a combination thereof.

5. The isolated anti-HIV antibody or antigen-binding portion thereof according to claim 4, wherein the heavy chain amino acid sequence further comprises substitutions of M428L and N434S.

6. The isolated anti-HIV antibody or antigen-binding portion thereof according to claim 5, wherein the light chain amino acid sequence comprises one or more of the following substitutions: T5D, T10D, S12D, S14D, I20D, and S22D; or, S61D, S63D, W65D, N70D, S72D, and S74D.

7. The isolated anti-HIV antibody or antigen-binding portion thereof according to any one of claims 1 to 6, wherein the heavy chain amino acid sequence comprises G54W and T68D, or a substitution of G54W.

8. The isolated anti-HIV antibody or antigen-binding portion thereof according to any one of claims 1 to 7, wherein the heavy chain amino acid sequence comprises any one amino acid sequence of SEQ ID NOs: 3 to 28 and SEQ ID NOs: 31 to 35.

9. The isolated anti-HIV antibody or antigen-binding portion thereof according to any one of claims 1 to 8, wherein the light chain amino acid sequence comprises any one amino acid sequence of SEQ ID NO: 2, SEQ ID NOs: 42 to 57, and SEQ ID NOs: 62 to 80.

10. The isolated anti-HIV antibody or antigen-binding portion thereof according to any one of claims 1 to 9, wherein the heavy chain amino acid sequence and the light chain amino acid sequence comprise the respective amino acid sequences shown in the sequence set of SEQ ID NO: 4 and SEQ ID NO:

2.

11. An isolated nucleic acid molecule comprising sequences encoding the heavy chain variable region and the light chain variable region of the anti-HIV antibody or antigen-binding portion thereof according to any one of claims 1 to 10.

12. A vector comprising the nucleic acid molecule according to claim 11.

13. A cultured cell comprising the vector according to claim 12.

14. Obtaining the cultured cell according to claim 13; Culturing the cultured cell in a medium under conditions that allow expression of the polypeptide encoded by the vector and assembly of the antibody or its fragment; and Purifying the antibody or its antigen-binding portion from the cultured cell or the medium of the cultured cell, A method for producing an anti-HIV antibody or antigen-binding portion thereof.

15. (i) At least one anti-HIV antibody or antigen-binding portion thereof according to any one of claims 1 to 10, the nucleic acid molecule according to claim 11, or the vector according to claim 12; and (ii) A pharmaceutically acceptable carrier, A pharmaceutical composition.

16. The pharmaceutical composition according to claim 15, further comprising an additional therapeutic agent.

17. The pharmaceutical composition according to claim 16, wherein the additional therapeutic agent comprises an antiviral agent or one or more additional antibodies.

18. The pharmaceutical composition according to claim 17, wherein the one or more additional antibodies comprise a second anti-HIV antibody or antigen-binding portion thereof, or a third antibody that binds to a third antigen.

19. The pharmaceutical composition according to claim 18, wherein the third antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-l), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), and CD160 (NK1).

20. The pharmaceutical composition according to claim 17, wherein the antiviral agent is selected from the group consisting of a non-nucleoside reverse transcriptase inhibitor, a protease inhibitor, an entry inhibitor or a fusion inhibitor, and an integrase inhibitor.

21. A pharmaceutical composition comprising at least one anti-HIV antibody or an antigen-binding portion thereof according to any one of claims 1 to 10, and used for preventing or treating HIV infection or an HIV-related disease.

22. The pharmaceutical composition according to claim 21, further comprising a second therapeutic agent.

23. The pharmaceutical composition according to claim 22, wherein the second therapeutic agent comprises an antiviral agent or one or more additional antibodies.

24. The pharmaceutical composition according to claim 23, wherein the one or more additional antibodies comprise a second anti-HIV antibody or an antigen-binding portion thereof, or a third antibody that binds to a third antigen.

25. The third antigen is selected from the group consisting of CD3, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16); CD89, CCR5, CD4, CD8, CD28, CD137, CTLA-4, gp41, killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, three Ig domains, and long cytoplasmic tail 1 (KIR3DL1), killer cell immunoglobulin-like receptor, two Ig domains, and long cytoplasmic tail 1 (KIR2DL1), killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3), killer cell lectin-like receptor Cl (KLRC1), killer cell lectin-like receptor C2 (KLRC2), killer cell lectin-like receptor C3 (KLRC3), killer cell lectin-like receptor C4 (KLRC4), killer cell lectin-like receptor D1 (KLRD1), killer cell lectin-like receptor K1 (KLRK1), natural cytotoxicity triggering receptor 3 (NCR3 or NKp30), natural cytotoxicity triggering receptor 2 (NCR2 or NK-p44), natural cytotoxicity triggering receptor 1 (NCR1 or NK-p46), CD226 (DNAM-1), cytotoxic and regulatory T cell molecule (CRTAM or CD355), signaling lymphocyte activation molecule family member 1 (SLAMF1), CD48 (SLAMF2), lymphocyte antigen 9 (LY9 or SLAMF3), CD244 (2B4 or SLAMF4), CD84 (SLAMF5), SLAM family member 6 (SLAMF6 or NTB-A), SLAM family member 7 (SLAMF7 or CRACC), CD27 (TNFRSF7), semaphorin 4D (SEMA4D or CD100), and CD160 (NK1). The pharmaceutical composition according to claim 24.

26. An HIV detection kit comprising a pharmaceutically acceptable dosage unit of a pharmaceutically effective amount of at least one isolated anti-HIV antibody according to any one of claims 1 to 10, or an antigen-binding portion thereof.

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