Arenavirus antibodies and uses thereof

Abstract

Disclosed herein are antibodies, including anti-GPl antibodies that cross-react with more than one New World arenavirus and methods of using the anti-GPl antibodies.

Description

[0001] Atorney Docket No. 117823-37620

[0002] ARENAVIRUS ANTIBODIES AND USES THEREOF

[0003] RELATED APPLICATIONS

[0004] The instant application claims priority to U.S. Provisional Application No. 63 / 747,710, filed on January 21, 2025. The entire contents of the foregoing application are expressly incorporated by reference herein.

[0005] SEQUENCE LISTING

[0006] The instant application contains a Sequence Listing which has been submited electronically in Sequence Listing XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on January 13, 2026 is named 117823-37620_SL.xml and is 35,865 bytes in size.

[0007] GOVERNMENT SUPPORT

[0008] This invention was made with government support under OD023084 awarded by National Institutes of Health (NIH). The government has certain rights in this invention.

[0009] BACKGROUND

[0010] Arenaviruses are enveloped RNA viruses that cause lethal hemorrhagic fevers when they are transmited from rodents to humans. They primarily circulate in South America. There are no Food and Drug Administration (FDA) -approved arenavirus vaccines and current anti-arenaviral therapy is limited to an off-label use of the antiviral drug ribavirin that is only partially effective and can cause significant side effects. Ribavirin may be effective in the treatment of Lassa fever only if administered early in the course of illness. Accordingly, there remains a need in the art for compositions that can be used for the treatment of arenaviruses and hemorrhagic fevers.

[0011] SUMMARY

[0012] The present invention is based on the discovery of antibodies with broad activity against one or more arenaviruses. In particular, the disclosed antibodies were isolated from Argentine hemorrhagic fever (AHF) survivors and were shown to display broad activities against two or more arenaviruses, including Junin virus (JUNV), the GP1 of Machupo virus (MACV), Sabia virus (SBAV), SBAV-like virus (SBAV-like), and / or Chapare virus (CHAPV) pseudotype. The exemplary data disclosed herein show that the antibodies contain CX4-5CY motif, of which the paired cysteine residues can form disulfide bridge that supports the extended length of the CDRH3, allowing for flexible but stabilized insertion into the GP1 pocket. These antibodies represent promising candidates for therapeutic development and provide a framework for the development of vaccines that induce broadly neutralizing antibody responses.Atorney Docket No. 117823-37620

[0013] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 4 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 8.

[0014] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 3 and a light chain variable region comprising a CDR2 having the amino acid sequence GAS.

[0015] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 2 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 6.

[0016] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 12 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 16.

[0017] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 11 and a light chain variable region comprising a CDR2 having the amino acid sequence EVS.

[0018] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 14.

[0019] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 20 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0020] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising a CDR2 having the amino acid sequence GAS.

[0021] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 18 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 22.

[0022] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variableAtorney Docket No. 117823-37620

[0023] region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 28 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 32.

[0024] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 27 and a light chain variable region comprising a CDR2 having the amino acid sequence DAS.

[0025] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 26 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 30.

[0026] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 3, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 2, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 8, a CDR2 domain comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 6.

[0027] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 11, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 domain comprising the amino acid sequence EVS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 14.

[0028] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 20, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 19, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 18, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 24, a CDR2 domain comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 22.

[0029] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 28, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 27, and a CDR1 domain comprising the amino acid sequence of SEQ IDAtorney Docket No. 117823-37620

[0030] NO: 26, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 32, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 31, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 30.

[0031] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises an amino acid sequence set forth in SEQ ID NO: 1, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 5, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 5.

[0032] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 1 and a light chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0033] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 9, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 9, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 13, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13.

[0034] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 13.

[0035] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21.

[0036] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 21.

[0037] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 90%,Atorney Docket No. 117823-37620

[0038] 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 29.

[0039] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 29.

[0040] In some embodiments, the GP1 is the GP1 of Junin virus (JUNV) and / or the GP1 of Machupo virus (MACV). In some embodiments, the GP1 is the GP1 of JUNV. In some embodiments, the GP1 is the GPl of MACV.

[0041] In some embodiments, the GP1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 33-40, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 33-40.

[0042] In some embodiments, the GP1 comprises the amino acid sequence set forth in SEQ ID NO: 33 or 34, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 33 or 34.

[0043] In some embodiments, the GP1 comprises the amino acid sequence set forth in SEQ ID NO: 35 or 36, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 35 or 36.

[0044] In some embodiments, the GP1 comprises an epitope comprising an asparagine (D), a serine (S), and a positive charged amino acid residue. In some embodiments, the positive charged amino acid residue is selected from the group consisting of lysine (K) and arginine (R).

[0045] In some embodiments, the GP1 comprises an epitope comprising a serine (S), an asparagine (D), an isoleucine (I), a valine (V), a tyrosine (Y), an lysine (K), and a glutamic acid (E).

[0046] In some embodiments, the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or all, of amino acid residues Sill, DI 13, DI 14, 1115, Al 16, VI 17, LI 19, Y122, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

[0047] In some embodiments, the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all, of amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 19, Y122, K169, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

[0048] In some embodiments, the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all, of amino acid residues Sill, DI 13, DI 14, 1115, Al 16, VI 17, LI 18, LI 19, E121, Y122, D123, R165, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

[0049] In some embodiments, the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or all, of amino acid residues S97, F98, Rill, VI 12, SI 13, DI 14, 1115, SI 16, VI 17, Ml 19, Y122, R165, K169, K170, E171, N174, T216, N218, F226, and Y228 of the amino acid sequence set forth in SEQ ID NO: 35.Atorney Docket No. 117823-37620

[0050] In some embodiments, the heavy chain variable region comprises a CDR3 that contributes at least about 70% of the contact area between the isolated antibody, or antigen binding portion thereof, and GP 1.

[0051] In some embodiments, the heavy chain variable region comprises a CDR3 that contributes at least about 75% of the contact area between the isolated antibody, or antigen binding portion thereof, and GP 1.

[0052] In some embodiments, the antibody, or antigen binding fragment thereof, competes for binding with transferrin receptor 1 (TfRl).

[0053] In some embodiments, the isolated antibody, or antigen binding fragment thereof, cross-reacts with two or more arenaviruses selected from the group consisting of JUNV, MACV, Sabia virus (SBAV), SBAV-like virus (SBAV-like), and Chapare virus (CHAPV).

[0054] In some embodiments, the isolated antibody, or antigen binding fragment thereof, cross-reacts with two or more arenaviruses selected from the group consisting of JUNV, SBAV, and SBAV-like. In some embodiments, the isolated antibody, or antigen binding fragment thereof, further cross-reacts with at least one of MACV and CHAPV.

[0055] In some embodiments, the antibody, or antigen binding fragment thereof, neutralizes JUNV at an IC50 of about 100 ng / mL or lower, in vitro.

[0056] In some embodiments, the antibody, or antigen binding fragment thereof, neutralizes JUNV at an IC50 of about 50 ng / mL or lower, in vitro.

[0057] In some embodiments, the antibody, or antigen binding fragment thereof, comprises a Fab, Fab2, or scFv.

[0058] In some embodiments, the antibody, or antigen binding fragment thereof, comprises a constant region, an Fc region, or at least one domain thereof.

[0059] In some embodiments, the constant region or Fc region comprises a mutation which impairs at least one effector function,

[0060] In some embodiments, the effector function is selected from the group consisting of FcR binding, complement binding, glycosylation, complement-dependent cytotoxicity (CDC), and antibody-dependent cellular cytotoxicity (ADCC).

[0061] In some embodiments, the constant or Fc region is a human constant or Fc region.

[0062] In some embodiments, the human constant or Fc region is selected from the group consisting of a human IgGl constant region , a human IgG2 constant region, a human IgG3 constant region, a human IgG4 constant region, and a human Fc constant region.

[0063] In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a chimeric antibody.

[0064] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding fragment thereof, which binds the same epitope as the isolated antibody disclosed herein, or antigen binding fragment thereof.Atorney Docket No. 117823-37620

[0065] In some aspects of the invention, the present disclosure provides an isolated nucleic acid encoding an antibody disclosed herein, or antigen binding portion thereof.

[0066] In some aspects of the invention, the present disclosure provides a pharmaceutical composition comprising the antibody disclosed herein, or antigen binding portion thereof , and a pharmaceutically acceptable carrier.

[0067] In some aspects of the invention, the present disclosure provides a method of treating an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with said infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an antibody or antigen binding fragment thereof disclosed herein, or a pharmaceutical composition disclosed herein, thereby treating the subject.

[0068] In some embodiments, said condition, symptom, disease, or disorder comprises at least one of fever, malaise, headache, conjunctivitis, nausea, vomiting, diarrhea, thrombocytopenia, bleeding, neurological disorders, petechiae (such as petechiae in oral mucosa, chest, arms, or axillary region), ataxia, tremors, seizures, and coma.

[0069] In some aspects of the invention, the present disclosure provides a method of preventing infection by one or more arenaviruses in a subject in need thereof, comprising administering to the subject a prophylactically effective amount of an antibody or antigen binding fragment disclosed herein, or a pharmaceutical composition disclosed herein, thereby preventing infection by one or more arenaviruses in the subject.

[0070] In some aspects of the invention, the present disclosure provides a method of inducing an immune response against one or more arenaviruses in a subject in need thereof, comprising administering an effective amount of an antibody or antigen binding fragment thereof disclosed herein, or a pharmaceutical composition disclosed herein, hereby inducing an immune response against one or more arenaviruses in the subject.

[0071] In some embodiments, said immune response elicits immunoprotection against the one or more arenaviruses.

[0072] In some aspects of the invention, the present disclosure provides a method of inhibiting or blocking infection of susceptible cells by one or more arenaviruses in a subject in need thereof, comprising administering an effective amount of an antibody or antigen binding fragment thereof disclosed herein, or a pharmaceutical composition disclosed herein, thereby inhibiting or blocking infection of susceptible cells by one or more arenaviruses in the subject.

[0073] In some aspects of the invention, the present disclosure provides a method of preventing the onset of a hemorrhagic fever in a subject infected by one or more arenaviruses, or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in a subject for a subject infected by one or more arenaviruses, comprising administering to the subject a prophylactically or therapeutically effective amount of an antibody or antigen binding fragment thereof disclosed herein,Atorney Docket No. 117823-37620

[0074] or a pharmaceutical composition disclosed herein, thereby preventing the onset of a hemorrhagic fever or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in the subject.

[0075] In some embodiments, the one or more arenaviruses comprise Junin virus (JUNV), Machupo virus (MACV), Sabia virus (SBAV), SBAV-like virus (SBAV-like), or Chapare virus (CHAPV).

[0076] In some embodiments, the one or more arenaviruses comprise JUNV, and one or more of MACV, SBAV, SBAV-like, and CHAPV.

[0077] In some embodiments, the one or more arenaviruses comprise JUNV, SBAV, and SBAV-like, and one or more of MACV and CHAPV.

[0078] In some embodiments, the hemorrhagic fever is selected from the group consisting of Argentine hemorrhagic fever, Bolivian hemorrhagic fever, Brazilian hemorrhagic fever, and hemorrhagic fever caused by CHAPV.

[0079] In some embodiments, the subject is a human subject.

[0080] In some embodiments, the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered intravenously, intramuscularly, or subcutaneously.

[0081] In some embodiments, the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject at a dose of about 100 mg to about 2000 mg, about 200 mg to about 1500 mg, about 300 mg to about 600 mg, about 500 mg to about 1200 mg, or about 300 mg to about 1200 mg.

[0082] In some embodiments, the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject at a dose of about 0.1 mg / kg to 1000 mg / kg, about 1 mg / kg to about 500 mg / kg, about 1 mg / kg to about 200 mg / kg, about 1 mg / kg to about 100 mg / kg, or about 0.1 mg / kg to about 50 mg / kg.

[0083] In some embodiments, the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject daily, weekly, monthly, every two months, every three months, or every six months.

[0084] In some embodiments, the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject in combination with an anti -arenavirus agent or an anti-arenavirus therapy.

[0085] In some aspects of the invention, the present disclosure provides a method of producing the antibody or antigen binding portion thereof disclosed herein, the method comprising expressing the antibody, or antigen binding portion thereof, in a recombinant cell, and isolating the antibody, or antigen binding portion thereof, from the cell.

[0086] In some embodiments, the method disclosed herein further comprises formulating the antibody, or antigen binding portion thereof, isolated from the cell into a pharmaceutical composition.

[0087] In some aspects, the present disclosure provides a kit comprising the antibody or antigen binding portion thereof disclosed herein, or the pharmaceutical composition disclosed herein.Atorney Docket No. 117823-37620

[0088] In some aspects, the present disclosure provides a syringe comprising the antibody or antigen binding portion thereof disclosed herein, or the pharmaceutical composition disclosed herein.

[0089] In some aspects, the present disclosure provides an intravenous bag comprising the antibody or antigen binding portion thereof disclosed herein, or the pharmaceutical composition disclosed herein.

[0090] BRIEF DESCRIPTION OF THE DRAWINGS

[0091] Figures 1A and IB show cross-reactivity of New World arenaviruses by Argentine hemorrhagic fever (AHF)-derived CX4-5CY motif-containing monoclonal antibodies. Figure 1A shows neutralizing IC50 curves for motif-containing antibodies pre-incubated with JUNV pseudotype. The left heatmap in Figure IB summarizes the percent neutralization of New World arenavirus pseudotypes by 13 CX4-5CY motif-containing antibodies. The number of pseudoviruses neutralized by each antibody are indicated on the botom X axis, and the total number of antibodies that neutralize each pseudotype are indicated on the right Y axis. The right heatmap summarizes the percent neutralization of New World arenavirus pseudotypes by 13 non-motif containing antibodies. The number of pseudoviruses neutralized by each antibody are indicated on the botom X axis, and the total number of antibodies that neutralize each pseudotype are indicated on the right Y axis.

[0092] Figures 2A and 2B show cross-neutralizing IC50 profiles of the tested antibodies. Figure 2A displays curves showing neutralization of murine leukemia virus pseudotypes by CX4-5CY motifcontaining hits from initial cross-neutralization screen. Cross-neutralizing antibodies are shown in top left (AHF1 B7), center middle (AHF2 C5), center right (AHF2 C8), and botom left (AHF3 E8.2). Figure 2B displays curves neutralization of murine leukemia virus pseudotypes by non-CX4-5CY motif-containing hits from initial cross-neutralization screen. For both panels: only hits validated to have >10% neutralization of New World arenavirus pseudotypes at the highest concentration tested (316 pg ml;1) are shown.

[0093] Figures 3A, 3B, and 3C show cross-neutralization of JUNV and MACV subtypes by neutralizing antibodies. Figure 3 A shows crystal structure of antibody E8.2 in complex with JUNV GP1. Figure 3B shows crystal structure of AHF1 B7 Fab in complex with MACV GP1. Figure 3C shows crystal structure of AHF1 B7 Fab in complex with JUNV GP1.

[0094] DETAILED DESCRIPTION

[0095] Various aspects of the disclosure relate to broadly cross-reactive antibodies and antibody fragments binding glycoprotein subunit 1 (GP1) of one or more arenaviruses, and pharmaceutical compositions thereof, as well as nucleic acids, recombinant expression vectors and host cells for making such antibodies and fragments. Also provided herein are methods of using the antibodies described herein to detect GP1 of one or more arenaviruses, to bind to and inhibit GP1 of one or more arenaviruses expressing cells, to inhibit, prevent, treat, and / or ameliorate infection by one or moreAtorney Docket No. 117823-37620

[0096] arenaviruses, in vivo, and / or to prevent and / or treat arenavirus-associated disorders, e.g., hemorrhagic fever, including, but not limited to, Argentine hemorrhagic fever, Bolivian hemorrhagic fever, Brazilian hemorrhagic fever, or hemorrhagic fever caused by CHAPV.

[0097] I. Definitions

[0098] In order that the invention may be more readily understood, certain terms are first defined. In addition, it should be noted that whenever a value or range of values of a parameter are recited, it is intended that values and ranges intermediate to the recited values are also intended to be part of this invention.

[0099] It is to be understood that this invention is not limited to the particular methodology, protocols, cell lines, animal species or genera, and reagents described, as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which will be limited only by the appended claims. As used herein the singular forms "a", "and", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and reference to "the protein" includes reference to one or more proteins and equivalents thereof known to those skilled in the art, and so forth. All technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this invention belongs unless clearly indicated otherwise.

[0100] The terms “antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1)”, “a receptor binding glycoprotein subunit 1 (GP1) antibody” or “anti-GP1 antibody”, used interchangeably herein, refer to an antibody that specifically binds to GP1, e.g., a GP1 of an arenavirus. An antibody “which binds” an antigen of interest, i.e., GP1, is one capable of binding that antigen with sufficient affinity such that the antibody is useful in targeting a cell expressing the antigen. In a preferred embodiment, the antibody specifically binds to a GP1 of an arenavirus (e.g., GP1 of Junin virus (JUNV)). Examples of anti-GPl antibodies are disclosed in the Examples below. Unless otherwise indicated, the term “anti-GPl antibody” is meant to refer to an antibody which binds to wild type GP1 of an arenavirus (e.g., GP1 of Junin virus (JUNV), GP1 of Machupo virus (MACV), GP1 of Sabia virus (SBAV), GP1 of SBAV-like virus (SBAV -like), or GP1 of Chapare virus (CHAPV)), a variant, or an isoform of GP1.

[0101] The term “GP1” is used to describe the GP1 subunit of any arenavirus. In some embodiments, the term “GP1” is used to describe the GP1 subunit of any arenavirus (e.g., New World arenavirus).

[0102] An exemplary amino acid sequence of wild type GP1 of JUNV, which contains 485 amino acids, is provided below as SEQ ID NO: 33. An exemplary amino acid sequence of wild type GP1 of JUNV, which contains 193 amino acids, is provided below as SEQ ID NO: 34. An exemplary antibody-binding domain of the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, II, or all, of amino acids Sill, D113, D114, 1115, A116, V117, L119, Y122, K169, E171, K216, andAtorney Docket No. 117823-37620

[0103] Q218, or any combination thereof of SEQ ID NO: 33. An exemplary antibody-binding domain of the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or all, of amino acids Sil l, DI 13, DI 14, 1115, Al 16, VI 17, LI 18, LI 19, E121, Y122, D123, R165, E171, K216, and Q218, or any combination thereof of SEQ ID NO: 33.

[0104] > glycoprotein precursor [Mammarenavirus juninense] (GenBank: BAA00964.2) (SEQ ID NO: 33) MGQFISFMQEIPTFLQEALNIALVAVSLIAIIKGVVNLYKSGLFQFFVFLALAGRSCTE EAFKIGLHTEFQT VSF SMVGLF SNNPHDLPLLCTLNKSHL YIKGGNASFKISFDDIAVL LPEYDVIIQHPADMSWCSKSDDQIWLSQWFMNAVGHDWYLDPPFLCRNRTKTEGFI FQVNTSKTGINENYAKKFKTGMHHLYREYPDSCLDGKLCLMKAQPTSWPLQCPLDH VNTLHFLTRGKNIQLPRRSLKAFFSWSLTDSSGKDTPGGYCLEEWMLVAAKMKCFG NTAVAKCNLNHDSEFCDMLRLFDYNKNAIKTLNDETKKQVNLMGQTINALISDNLL MKNKIRELMS VP YCNYTKFWYVNHTLSGQHSLPRCWLIKNNSYLNISDFRND WILES DFLISEMLSKEYSDRQGKTPLTLVDICFWSTVFFTASLFLHLVGIPTHRHIRGEACPLP HRLNSLGGCRCGI<YPNL1<I<PTVWRRGH

[0105] >JUNV virus (MC2 strain; GenBank: D10072.2) (SEQ ID NO: 34) EEAFKIGLHTEFQTVSFSMVGLFSNNPHDLPLLCTLNKSHLYIKGGNASFKISFDDIAVLLPEY DVIIQHPADMSWCSKSDDQIWLSQWFMNAVGHDWYLDPPFLCRNRTKTEGFIFQVNTSKTGI NENYAKKFKTGMHHLYREYPDSCLDGKLCLMKAQPTSWPLQCPLDHVNTLHFLTRGKNIQL PRRSLK

[0106] An exemplary amino acid sequence of wild type GP 1 of MACV, which contains 496 amino acids, is provided below as SEQ ID NO: 35. An exemplary amino acid sequence of wild type GP1 of MACV, which contains 262 amino acids, is provided below as SEQ ID NO: 36. An exemplary antibody-binding domain of the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or all, of amino acids S97, F98, R111, VI 12, SI 13, DI 14, Il 15, SI 16, VI 17, Ml 19, Y122, R165, K169, K170, E171, N174, T216, N218, F226, and Y228, or any combination thereof of SEQ ID NO: 35.

[0107] > glycoprotein precursor [Mammarenavirus machupoense] (Genbank: AAN05425.1) (SEQ ID NO: 35) MGQLISFFQEIPVFLQEALNIALVAVSLIAVIKGIINLYKSGLFQFIFFLLLAGRSCSDGTFKIGL HTEFQSVTLTMQRLLANHSNELPSLCMLNNSFYYMRGGVNTFLIRVSDISVLMKEYDVSIYEP EDLGNCLNKSDSSWAIHWFSNALGHDWLMDPPMLCRNKTKKEGSNIQFNISKADDARVYG KKIRNGMRHLFRGFHDPCEEGKVCYLTINQCGDPSSFDYCGVNHLSKCQFDHVNTLHFLVRS KTHLNFERSLKAFFSWSLTDSSGKDMPGGYCLEEWMLIAAKMKCFGNTAVAKCNQNHDSE FCDMLRLFDYNKNAIKTLNDESKKEINLLSQTVNALISDNLLMKNKIKELMSIPYCNYTKFW YVNHTLTGQHTLPRCWLIRNGSYLNTSEFRNDWILESDHLISEMLSKEYAERQGKTPITLVDI CFWSTIFFTASLFLHLVGIPTHRHLKGEACPLPHKLDSFGGCRCGKYPRLKKPTIWHKRH

[0108] >Machupo virus (Carvallo strain; Genbank: AAN05425.1) (SEQ ID NO: 36) MGQLISFFQEIPVFLQEALNIALVAVSLIAVIKGIINLYKSGLFQFIFFLLLAGRSCSDGTFKIGL HTEFQSVTLTMQRLLANHSNELPSLCMLNNSFYYMRGGVNTFLIRVSDISVLMKEYDVSIYEP EDLGNCLNKSDSSWAIHWFSNALGHDWLMDPPMLCRNKTKKEGSNIQFNISKADDARVYG KKIRNGMRHLFRGFHDPCEEGKVCYLTINQCGDPSSFDYCGVNHLSKCQFDHVNTLHFLVRSAtorney Docket No. 117823-37620

[0109] KTHLNFERSLK

[0110] An exemplary amino acid sequence of wild type GP1 of Guanarito vims, which contains 187 amino acids, is provided below as SEQ ID NO: 37.

[0111] >Guanarito virus (Genbank: AAN05423.1) (SEQ ID NO: 37) FKVGHHTNFESFTVKLGGVFHELPSLCRVNNSYSLIRLSHNSNQALSVEYVDVHPVLCSSSPTI LDNYTQCIKGSPEFDWILGWTIKGLGHDFLRDPRICCEPKKTTNAEFTFQLNLTDSPETHHYR SKIEVGIRHLFGNYITNDSYSKMSVVMRNTTWEGQCSNSHVNTLRFLVKNAGYLVGRKPL

[0112] An exemplary amino acid sequence of wild type GP1 of Sabia vims, which contains 196 amino acids, is provided below as SEQ ID NO: 38.

[0113] >Sabia virus (Genbank: AFA53089.1) (SEQ ID NO: 38) FRIGRSTELQNITFDMLKVFEDHPTSCMVNHSTYYVHENKNATWCLEVSVTDVTLLMAEHD RQVLNNLSNCVHPAVEHRSRMVGLLEWIFRALKYDFNHDPTPLCQKQTSTVNETRVQINITE GFGSHGFEDTILQRLGVLFGSRIAFSNIQDLGKKRFLLIRNSTWKNQCEMNHVNSMHLMLAN AGRSSGSRRPL

[0114] An exemplary amino acid sequence of wild type GP1 of Sabia-like vims, which contains 196 amino acids, is provided below as SEQ ID NO: 39.

[0115] >Sabia-like virus (Genbank: QHW12249.1) (SEQ ID NO: 39) FRIGRSTELQNITFDMLKVFEDHPTSCMVSHSTYYVHENRNATWCLEVSVTDVTLLMVEHDR QVLNNLSNCVHPAVKHRSRMVGLLEWIFRALKYDFNHDPTPLCQKETSTVNETRVQINITEG YWSHGFKDTILQRLGVLFGSRIAFSDLQKIGPKTFLLIRNTTWSNQCVMNHVNSMHLMLANA GRSFSSRRPL

[0116] An exemplary amino acid sequence of wild type GP1 of Chapare vims, which contains 192 amino acids, is provided below as SEQ ID NO: 40.

[0117] >Chapare virus (Genbank: ABY87068.1) (SEQ ID NO: 40) FKIGRSTELQNITINMLKVFEDHPISCTVNKTLYYIRESENATWCVEIAALDMSVLLSPHDPRV MGNLSNCVHPDIKHRSELLGLLEWILRALKYDFLNYPPLLCEKVTSSVNETRIQINVSDSAGS HDFKETMLQRLAILFGTKLMFDKTPKQFIVIRNQTWVNQCKSNHVNTLHLMMANAGHAVK LRRLQ

[0118] In some embodiments, the term “GP1” or “GP1 of an arenavirus” as used herein encompasses a polypeptide sequence selected from the group consisting of SEQ ID NOs: 33-40, and any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to a polypeptide sequence selected from the group consisting of SEQ ID NOs: 33-40, or a fragment thereof.Atorney Docket No. 117823-37620

[0119] In some embodiments, the GP1 of JUNV comprises the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 33.

[0120] In some embodiments, the GP1 of JUNV comprises the amino acid sequence set forth in SEQ ID NO: 34, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 34.

[0121] In some embodiments, the GP1 of MACV comprises the amino acid sequence set forth in SEQ ID NO: 35, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 35.

[0122] In some embodiments, the GP1 of MACV comprises the amino acid sequence set forth in SEQ ID NO: 36, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 36.

[0123] In some embodiments, the GP1 of Guanarito virus comprises the amino acid sequence set forth in SEQ ID NO: 37, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 37.

[0124] In some embodiments, the GP1 of SBAV comprises the amino acid sequence set forth in SEQ ID NO: 38, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 38.

[0125] In some embodiments, the GP1 of SBAV-like virus comprises the amino acid sequence set forth in SEQ ID NO: 39, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 39.

[0126] In some embodiments, the GP1 of CHAPV comprises the amino acid sequence set forth in SEQ ID NO: 40, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 40.

[0127] The terms “specific binding” or “specifically binding”, as used herein, in reference to the interaction of an antibody or antigen binding portion thereof disclosed herein with a second chemical species, mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody or antigen binding portion thereof is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody or an antigen binding portion thereof, will reduce the amount of labeled A bound to the antibody or the antigen binding portion thereof.

[0128] In some embodiments, the phrase “specifically binds to GP1” or “specific binding to GP1”, as used herein, refers to the ability of an anti-GPl antibody, or an antigen binding portion thereof, to interact with GP1 (e.g., GP1 of junin virus (JUNV), Machupo virus (MACV), Sabia virus (SBAV), SBAV-like virus (SBAV -like), and / or Chapare virus (CHAPV)) with a dissociation constant (KD) ofAtorney Docket No. 117823-37620

[0129] about 2,000 nM or less, about 1,000 nM or less, about 500 nM or less, about 200 nM or less, about 100 nM or less, about 75 nM or less, about 25 nM or less, about 21 nM or less, about 12 nM or less, about 11 nM or less, about 10 nM or less, about 9 nM or less, about 8 nM or less, about 7 nM or less, about 6 nM or less, about 5 nM or less, about 4 nM or less, about 3 nM or less, about 2 nM or less, about 1 nM or less, about 0.5 nM or less, about 0.3 nM or less, about 0.1 nM or less, about 0.01 nM or less, or about 0.001 nM or less. In some embodiments, the phrase “specifically binds to GP1” or “specific binding to GP1”, as used herein, refers to the ability of an anti-GPl antibody or antigen binding portion thereof to interact with GP 1 with a dissociation constant (KD) of between about 1 pM (0.001 nM) to 2,000 nM, between about 500 pM (0.5 nM) to 1,000 nM, between about 500 pM (0.5 nM) to 500 nM, between about 1 nM) to 200 nM, between about 1 nM to 100 nM, between about 1 nM to 50 nM, between about 1 nM to 20 nM, or between about 1 nM to 5 nM. In one embodiment, KD is determined by surface plasmon resonance or Bio-Layer Interferometry, or by any other method known in the art. Bio-Layer Interferometry refers to an optical phenomenon that allows for the analysis of real-time biospecific interactions by measuring the interference paterns of reflected white light, for example using the Octet™ system (ForteBio, Pall Corp. Fremont, CA). For further description of the Octet™ system, see Li, B et al. (2011) J. Pharm. Biomed. Anal. 54(2):286-294 and Abdiche, Y.N., et al. (2009) Anal. Biochem. 386(2): 172-180, the contents of which are incorporated herein by reference.

[0130] The term “antibody” broadly refers to an immunoglobulin (Ig) molecule, generally comprised of four polypeptide chains, two heavy (H) chains and two light (L) chains, or any functional fragment, mutant, variant, or derivative thereof, that retains the essential target binding features of an Ig molecule. Such mutant, variant, or derivative antibody formats are known in the art and non-limiting embodiments of which are discussed below.

[0131] In a full-length antibody, each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CHI, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY) and class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl and IgA2) or subclass.

[0132] The term “antigen binding portion” of an antibody (or simply “antibody portion”), as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., GP1 of an arenavirus). It has been shown that the antigen binding function of anAtorney Docket No. 117823-37620

[0133] antibody can be performed by fragments of a full-length antibody. Such antibody embodiments may also be bispecific, dual specific, or multi-specific formats; specifically binding to two or more different antigens. Examples of binding fragments encompassed within the term “antigen binding portion” of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CHI domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (W ard et al. , ( 1989) Nature 341 : 544-546, Winter et al. , PCT publication WO 90 / 05144 Al herein incorporated by reference), which comprises a single variable domain; and (vi) an isolated complementarity determining region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) roc. Natl. Acad. Sci. USA 85:5879-5883). Such single chain antibodies are also intended to be encompassed within the term “antigen binding portion” of an antibody. In certain embodiments, scFv molecules may be incorporated into a fusion protein. Other forms of single chain antibodies, such as diabodies are also encompassed. Diabodies are bivalent, bispecific antibodies in which VH and VL domains are expressed on a single polypeptide chain, but using a linker that is too short to allow for pairing between the two domains on the same chain, thereby forcing the domains to pair with complementary domains of another chain and creating two antigen binding sites (see e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, R.J., et al. (1994) Structure .1121-1123). Such antibody binding portions are known in the art (Kontermann and Dubel eds., Antibody Engineering (2001) Springer-Verlag. New York. 790 pp. (ISBN 3-540-41354-5).

[0134] The term “antibody construct” as used herein refers to a polypeptide comprising one or more the antigen binding portions disclosed herein linked to a linker polypeptide or an immunoglobulin constant domain. Linker polypeptides comprise two or more amino acid residues joined by peptide bonds and are used to link one or more antigen binding portions. Such linker polypeptides are well known in the art (see e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, R.J., et al. (1994) Structure .1121-1123). An immunoglobulin constant domain refers to a heavy or light chain constant domain. Antibody portions, such as Fab and F(ab')2 fragments, can be prepared from whole antibodies using conventional techniques, such as papain or pepsin digestion, respectively, of whole antibodies. Moreover, antibodies, antibody portions and immunoadhesion molecules can be obtained using standard recombinant DNA techniques, as described herein.

[0135] An “isolated antibody”, as used herein, is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody thatAtorney Docket No. 117823-37620

[0136] specifically binds GP1 is substantially free of antibodies that specifically bind antigens other than GP1). An isolated antibody that specifically binds GP1 may, however, have cross-reactivity to other antigens, such as GP1 molecules from other species. Moreover, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0137] The term “humanized antibody” refers to antibodies which comprise heavy and light chain variable region sequences from a nonhuman species (e.g., a mouse) but in which at least a portion of the VH and / or VL sequence has been altered to be more “human-like”, i. e. , more similar to human germline variable sequences. In particular, the term “humanized antibody” is an antibody or a variant, derivative, analog or fragment thereof which immunospecifically binds to an antigen of interest and which comprises a framework (FR) region having substantially the amino acid sequence of a human antibody and a complementary determining region (CDR) having substantially the amino acid sequence of a non-human antibody. As used herein, the term “substantially” in the context of a CDR refers to a CDR having an amino acid sequence at least 80%, preferably at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the amino acid sequence of a non-human antibody CDR. A humanized antibody comprises substantially all of at least one, and typically two, variable domains (Fab, Fab', F(ab')2, FabC, Fv) in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin (z. e. , donor antibody) and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence. Preferably, a humanized antibody also comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. In some embodiments, a humanized antibody contains both the light chain as well as at least the variable domain of a heavy chain. The antibody also may include the CHI, hinge, CH2, CH3, and CH4 regions of the heavy chain. In some embodiments, a humanized antibody only contains a humanized light chain. In other embodiments, a humanized antibody only contains a humanized heavy chain. In specific embodiments, a humanized antibody only contains a humanized variable domain of a light chain and / or humanized heavy chain.

[0138] The humanized antibody can be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA and IgE, and any isotype, including without limitation IgGl, IgG2, IgG3 and IgG4. The humanized antibody may comprise sequences from more than one class or isotype, and particular constant domains may be selected to optimize desired effector functions using techniques well-known in the art.

[0139] The terms “Kabat numbering,” “Kabat definitions,” and “Kabat labeling” are used interchangeably herein. These terms, which are recognized in the art, refer to a system of numbering amino acid residues which are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody, or an antigen binding portion thereof (Kabat et al. (1971) Ann. NY Acad, Sci. 190:382-391 and, Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable region, the hypervariable region ranges fromAtorney Docket No. 117823-37620

[0140] amino acid positions 31 to 35 for CDR1, amino acid positions 50 to 65 for CDR2, and amino acid positions 95 to 102 for CDR3. For the light chain variable region, the hypervariable region ranges from amino acid positions 24 to 34 for CDR1, amino acid positions 50 to 56 for CDR2, and amino acid positions 89 to 97 for CDR3.

[0141] As used herein, the term “CDR” refers to the complementarity determining region within antibody variable sequences. There are three CDRs in each of the variable regions of the heavy chain (HC) and the light chain (LC), which are designated CDR1, CDR2 and CDR3 (or specifically HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3), for each of the variable regions. The term “CDR set” as used herein refers to a group of three CDRs that occur in a single variable region capable of binding the antigen. The exact boundaries of these CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining the three CDRs. These CDRs may be referred to as Kabat CDRs. Chothia and coworkers (Chothia &Lesk, J. Mol. Biol. 196:901-917 (1987) and Chothia et al., Nature 342:877-883 (1989)) found that certain sub- portions within Kabat CDRs adopt nearly identical peptide backbone conformations, despite having great diversity at the level of amino acid sequence. These sub-portions were designated as LI, L2 and L3 or Hl, H2 and H3 where the “L” and the “H” designates the light chain and the heavy chains regions, respectively. These regions may be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs overlapping with the Kabat CDRs have been described by Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J Mol Biol 262(5):732-45 (1996)). Still other CDR boundary definitions may not strictly follow one of the above systems, but will nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. The methods used herein may utilize CDRs defined according to any of these systems, although preferred embodiments use Kabat or Chothia defined CDRs.

[0142] As used herein, the term “framework” or “framework sequence” refers to the remaining sequences of a variable region minus the CDRs. Because the exact definition of a CDR sequence can be determined by different systems, the meaning of a framework sequence is subject to correspondingly different interpretations. The six CDRs (CDR-L1, CDR-L2, and CDR-L3 of light chain and CDR-H1, CDR-H2, and CDR-H3 of heavy chain) also divide the framework regions on the light chain and the heavy chain into four sub-regions (FR1, FR2, FR3 and FR4) on each chain, in which CDR1 is positioned between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. Without specifying the particular sub-regions as FR1, FR2, FR3 or FR4, a framework region, as referred by others, represents the combined FR's within the variable region of a single,Atorney Docket No. 117823-37620

[0143] naturally occurring immunoglobulin chain. As used herein, a FR represents one of the four subregions, and FRs represents two or more of the four sub- regions constituting a framework region.

[0144] The framework and CDR regions of a humanized antibody need not correspond precisely to the parental sequences, e.g., the donor antibody CDR or the consensus framework may be mutagenized by substitution, insertion and / or deletion of at least one amino acid residue so that the CDR or framework residue at that site does not correspond to either the donor antibody or the consensus framework. In a preferred embodiment, such mutations, however, will not be extensive. Usually, at least 80%, preferably at least 85%, more preferably at least 90%, and most preferably at least 95% of the humanized antibody residues will correspond to those of the parental FR and CDR sequences. As used herein, the term “consensus framework” refers to the framework region in the consensus immunoglobulin sequence. As used herein, the term “consensus immunoglobulin sequence” refers to the sequence formed from the most frequently occurring amino acids (or nucleotides) in a family of related immunoglobulin sequences (See e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family of immunoglobulins, each position in the consensus sequence is occupied by the amino acid occurring most frequently at that position in the family. If two amino acids occur equally frequently, either can be included in the consensus sequence.

[0145] “Percent (%) amino acid sequence identity” with respect to a peptide or polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the specific peptide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. In one embodiment, the disclosure includes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an amino acid sequence set forth in any one of SEQ ID NOs: 1 to 32.

[0146] In one embodiment, the antibody, or antigen binding portion thereof, is capable of inducing antibody dependent cellular cytotoxicity (ADCC). In one embodiment, the antibody, or antigen binding portion thereof, is not capable of inducing ADCC.

[0147] In one embodiment, the antibody, or antigen binding portion thereof, is capable of inducing complement-dependent cytotoxicity (CDC). In one embodiment, the antibody, or antigen binding portion thereof, is not capable of inducing CDC.Atorney Docket No. 117823-37620

[0148] In one embodiment, the antibody, or antigen binding portion thereof, is capable of inducing ADCC and CDC. In one embodiment, the antibody, or antigen binding portion thereof, is not capable of inducing ADCC or CDC.

[0149] The term “multivalent antibody” is used herein to denote an antibody comprising two or more antigen binding sites. In certain embodiments, the multivalent antibody may be engineered to have the three or more antigen binding sites, and is generally not a naturally occurring antibody.

[0150] The term “multispecific antibody” refers to an antibody capable of binding two or more unrelated antigens.

[0151] The term “dual variable domain” or “DVD,” as used interchangeably herein, are antigen binding proteins that comprise two or more antigen binding sites and are tetravalent or multivalent binding proteins. Such DVDs may be monospecific, i.e., capable of binding one antigen or multispecific, i.e. capable of binding two or more antigens. DVD binding proteins comprising two heavy chain DVD polypeptides and two light chain DVD polypeptides are referred to a DVD Ig. Each half of a DVD Ig comprises a heavy chain DVD polypeptide, and a light chain DVD polypeptide, and two antigen binding sites. Each binding site comprises a heavy chain variable domain and a light chain variable domain with a total of 6 CDRs involved in antigen binding per antigen binding site. In one embodiment, the CDRs described herein are used in an anti-GPl DVD.

[0152] The term “activity” includes activities such as the binding specificity / affinity of an antibody for an antigen, for example, an anti-GPl antibody that binds to a GP1 antigen. In one embodiment, an anti-GPl antibody activity includes, but it not limited to, binding to GP1 of an arenavirus in vitro,' binding to GP1 of an arenavirus in vivo,' inhibiting GP1 binding to a virus cellular receptor (e.g., transferrin receptor 1 (TfRl)); inhibiting and / or preventing an arenavirus entry into a cell; inhibiting arenavirus replication; reducing a level of arenavirus; treating and / or preventing infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with said infection; preventing the onset of a hemorrhagic fever, or treating a hemorrhagic fever and / or symptoms of a hemorrhagic fever.

[0153] In one embodiment, the antibody, or antigen binding portion thereof, is capable of inducing antibody dependent cellular cytotoxicity (ADCC). In one embodiment, the antibody, or antigen binding portion thereof, is not capable of inducing ADCC.

[0154] In one embodiment, the antibody, or antigen binding portion thereof, is capable of inducing complement-dependent cytotoxicity (CDC). In one embodiment, the antibody, or antigen binding portion thereof, is not capable of inducing CDC.

[0155] In one embodiment, the antibody, or antigen binding portion thereof, is capable of inducing ADCC and CDC. In one embodiment, the antibody, or antigen binding portion thereof, is not capable of inducing ADCC or CDC.

[0156] The term “epitope” refers to a region of an antigen that is bound by an antibody or antibody fragment. In certain embodiments, epitope determinants include chemically active surface groupingsAtorney Docket No. 117823-37620

[0157] of molecules such as amino acids, sugar side chains, phosphoryl, or sulfonyl, and, in certain embodiments, may have specific three dimensional structural characteristics, and / or specific charge characteristics. In certain embodiments, an antibody is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and / or macromolecules.

[0158] The term “surface plasmon resonance”, as used herein, refers to an optical phenomenon that allows for the analysis of real-time biospecific interactions by detection of alterations in protein concentrations within a biosensor matrix, for example using the BIAcore system (Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, NJ). For further descriptions, see Jonsson, U., et al. (1993) Ann. Biol. Clin. 51:19-26; Jonsson, U., et al. (1991) Biotechniques 11:620-627; Johnsson, B., et al. (1995) J. Mol. Recognit. 8:125-131; and Johnnson, B., et al. (1991) Anal. Biochem. 198:268-277.

[0159] The term “ kon” or “ ka”, as used herein, is intended to refer to the on rate constant for association of an antibody to the antigen to form the antibody / antigen complex.

[0160] The term “kOfr” or “ ka”, as used herein, is intended to refer to the off rate constant for dissociation of an antibody from the antibody / antigen complex.

[0161] The term “KD”, as used herein, is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction. KD is calculated by ka / kd. In one embodiment, the antibodies of the invention have a KD of about 2,000 nM or less, about 1,000 nM or less, about 500 nM or less, about 200 nM or less, about 100 nM or less, about 75 nM or less, about 25 nM or less, about 21 nM or less, about 12 nM or less, about 11 nM or less, about 10 nM or less, about 9 nM or less, about 8 nM or less, about 7 nM or less, about 6 nM or less, about 5 nM or less, about 4 nM or less, about 3 nM or less, about 2 nM or less, about 1 nM or less, about 0.5 nM or less, about 0.3 nM or less, about 0.1 nM or less, about 0.01 nM or less, or about 0.001 nM or less.

[0162] The term “competitive binding”, as used herein, refers to a situation in which a first antibody competes with a second antibody, for a binding site on a third molecule, e.g., an antigen. In one embodiment, competitive binding between two antibodies is determined using FACS analysis.

[0163] The term “competitive binding assay” is an assay used to determine whether two or more antibodies bind to the same epitope. In one embodiment, a competitive binding assay is a competition fluorescent activated cell sorting (FACS) assay which is used to determine whether two or more antibodies bind to the same epitope by determining whether the fluorescent signal of a labeled antibody is reduced due to the introduction of a non-labeled antibody, where competition for the same epitope will lower the level of fluorescence.

[0164] The term “labeled antibody” as used herein, refers to an antibody, or an antigen binding portion thereof, with a label incorporated that provides for the identification of the binding protein, e.g., an antibody. Preferably, the label is a detectable marker, e.g., incorporation of a radiolabeled amino acid or atachment to a polypeptide of biotinyl moieties that can be detected by marked avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected byAtorney Docket No. 117823-37620

[0165] optical or colorimetric methods). Examples of labels for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (e.g.,3H14C,35S,90Y, "Tc,inIn,125I,1311,177Lu,166Ho, or153Sm); fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase); chemiluminescent markers; biotinyl groups; predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags); and magnetic agents, such as gadolinium chelates.

[0166] The term “administering” as used herein is meant to refer to the delivery of a substance (e.g., an anti-GPl antibody) to achieve a therapeutic objective (e.g., treatment, prevention, or reduction of infection by one or more arenaviruses or a condition, symptom, disease, or disorder associated with said infection; treatment, prevention, or amelioration of an arenavirus-associated hemorrhagic fever and / or the symptoms of an arenavirus-associated hemorrhagic fever). Modes of administration may be parenteral, enteral and topical. Parenteral administration is usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrastemal injection and infusion.

[0167] The term “combination therapy”, as used herein, refers to the administration of two or more therapeutic substances, e.g., an anti-GPl antibody or an antigen binding fragment thereof and an additional therapeutic agent. The additional therapeutic agent may be administered concomitant with, prior to, or following the administration of the anti-GPl antibody or an antigen binding fragment thereof. In one embodiment, the anti-GPl antibody or an antigen binding fragment thereof of the invention are administered in combination with an anti-arenavirus agent or an anti -arenavirus therapy.

[0168] As used herein, the term “effective amount” or “therapeutically effective amount” refers to the amount of a drug, e.g., an anti-GPl antibody or an antigen binding fragment thereof, which is sufficient to inhibit and / or prevent an arenavirus entry into a cell, reduce a level of an arenavirus, treat and / or prevent infection by an arenavirus, or treat a condition, symptom, disease, or disorder associated with said infection, prevent the onset of a hemorrhagic fever, or treat a hemorrhagic fever and / or the symptoms of a hemorrhagic fever, reduce or ameliorate the severity and / or duration of an infection or a disorder (e.g., hemorrhagic fever), or one or more symptoms thereof, prevent the advancement of a disorder (e.g., hemorrhagic fever), cause regression of a disorder (e.g., hemorrhagic fever), prevent the recurrence, development, onset or progression of one or more symptoms associated with a disorder (e.g., hemorrhagic fever), detect a virus and / or disorder, or enhance or improve the prophylactic or therapeutic effect(s) of another therapy (e.g., prophylactic or therapeutic agent). In some embodiments, the term “effective amount” or “therapeutically effective amount” refers to the amount of a drug, e.g., an anti-GPl antibody or an antigen binding fragment thereof, which is sufficient to eliminate an arenavirus from the subject or prevent the expansion of an arenavirus in the subject or eliminate or reduce the symptoms such as fever, malaise, headache, conjunctivitis, nausea,Atorney Docket No. 117823-37620

[0169] vomiting, diarrhea, thrombocytopenia, bleeding, neurological disorders, petechiae (such as petechiae in oral mucosa, chest, arms, or axillary region), ataxia, tremors, seizures, and coma.

[0170] As used herein, "treatment" is an approach for obtaining beneficial or desired clinical results. For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, one or more of the following: improvement in any aspect of an arenavirus-related condition such as fever, cough, or neurological disorders. For example, in the context of arenavirus infection treatment, this includes lessening severity, alleviation of fever, cough, neurological disorders, and other associated symptoms, reducing frequency of recurrence, increasing the quality of life of those suffering from the arenavirus-related symptoms, and decreasing dose of other medications required to treat the arenavirus-related symptoms. Other associated symptoms include, but are not limited to, malaise, headache, conjunctivitis, nausea, vomiting, diarrhea, thrombocytopenia, bleeding, neurological disorders, petechiae (such as petechiae in oral mucosa, chest, arms, or axillary region), ataxia, tremors, seizures, and coma.

[0171] "Reducing incidence" or “prophylaxis” or “prevention” means any of reducing severity for a particular disease, condition, symptom, or disorder (the terms disease, condition, and disorder are used interchangeably throughout the application). Reduction in severity includes reducing drugs and / or therapies generally used for the condition by, for example, reducing the need for, amount of, and / or exposure to drugs or therapies. Reduction in severity also includes reducing the duration, and / or frequency of the particular condition, symptom, or disorder (including, for example, delaying or increasing time to next episodic atack in an individual).

[0172] "Ameliorating" one or more symptoms of arenavirus infection-related conditions means a lessening or improvement of one or more symptoms of the condition, e.g., fever, malaise, headache, conjunctivitis, nausea, vomiting, diarrhea, thrombocytopenia, bleeding, neurological disorders, petechiae (such as petechiae in oral mucosa, chest, arms, or axillary region), ataxia, tremors, seizures, and coma as compared to not administering an anti-GPl antibody. "Ameliorating" also includes shortening or reduction in duration of a symptom.

[0173] Various aspects of the invention are described in further detail in the following subsections.

[0174] II. Antibodies that binds to GP1 of Arenavirus

[0175] The present invention provides antibodies, or antigen binding fragments thereof, that display broad activity against one or more arenaviruses (e.g., two or more arenaviruses, three or more arenaviruses, four or more arenaviruses, or five or more arenaviruses).Atorney Docket No. 117823-37620

[0176] Arenaviruses are a group of enveloped, bi-segmented RNA-containing viruses with unique ambisense genomic organization. Nearly all of the arenavirus members can cause acute or persistent infections of the infected hosts. The family Arenaviridae is divided into Old World and New World complexes. The Old World arenavirus complex includes, but not limited to, Lymphocytic choriomeningitis virus (LCMV), Lassa virus (LASV), Lujo virus, Kodoko virus (KODV), Mobala virus (MOBV), Ippy virus (IPPYV), Mopeia virus (MOPV), and Morogoro virus (MORV). The New World arenaviruses can be further divided into four clades: A, B, C, and A / Rec (Clade D). Clade A includes, but not limited to, Allpahuayo virus (AALV), Flexal virus (FLEV), Parana virus (PRAV), Pichinde virus (PICHV), Pirital virus (PIRV); Clade B includes, but not limited to, Chapare virus (CHAPV), Sabia virus (SBAV), Cupixi virus (CPXV), Guanarito virus (GTOV), Amapari virus (AMAV), Junin virus (JUNV), Machupo virus (MACV), and Tacaribe virus (TCRV); Clade C includes, but not limited to, Oliveros virus (OLVV) and Latino virus (LATV); Clade A / Rec include, but not limited to, Whitewater Arroyo virus (WWAV), Tamiami virus (TMMV), Bear Canyon virus (BCNV), Skinner Tank virus (SKTV), and Catarina virus (CTNV). JUNV, MACV, GTOV, and SABV are the causative agents of Argentine Hemorrhagic Fever (AHF), Bolivian Hemorrhagic Fever (BHF), Venezuelan Hemorrhagic Fever (VHF), and Brazilian Hemorrhagic Fever (BzHF), respectively.

[0177] In some aspects, disclosed herein are humanized anti-GP 1 antibodies, or antigen binding portions thereof, that bind to a GP1 of an arenavirus. In another aspect, disclosed herein are human anti-GP 1 antibodies, or antigen binding portions thereof, that bind to a GP1 of an arenavirus. In some embodiments, the GP1 is the GP1 of an Old World arenavirus. Accordingly, in some embodiments, the antibody or antigen binding portion thereof disclosed herein binds to GP1 of one or more arenaviruses, wherein the GP1 is selected from the group consisting of the GP1 of Lymphocytic choriomeningitis virus (LCMV), the GP1 of Lassa virus (LASV), the GP1 of Lujo virus, the GP1 of Kodoko virus (KODV), the GP1 of Mobala virus (MOBV), the GP1 of Ippy virus (IPPYV), the GP1 of Mopeia virus (MOPV), and the GP1 of Morogoro virus (MORV)).Atorney Docket No. 117823-37620

[0178] In some embodiments, the GP1 is the GP1 of a New World arenavirus (e.g. , a GP1 of a New World arenavirus in Clade A, a GP1 of a New World arenavirus in Clade B, a GP1 of a New World arenavirus in Clade C, or a GP1 of a New World arenavirus in Clade A / Rec). Accordingly, in some embodiments, the antibody or antigen binding portion thereof disclosed herein binds to GP 1 of one or more arenaviruses, wherein the GP1 is selected from the group consisting of the GP1 of Allpahuayo virus (AALV), the GP1 of Flexal virus (FLEV), the GP1 of Parana virus (PRAV), the GP1 of Pichinde virus (PICHV), the GP1 of Pirital virus (PIRV), the GP1 of Chapare virus (CHAPV), the GP1 of Sabia virus (SBAV), the GP1 of Cupixi virus (CPXV), the GP1 of Guanarito virus (GTOV), the GP1 of Amapari virus (AMAV), the GP1 of Junin virus (JUNV), the GP1 of Machupo virus (MACV), the GP1 of Tacaribe virus (TCRV), the GP1 of Oliveros virus (OLVV), the GP1 of Latino virus (LATV)), the GP1 of Whitewater Arroyo virus (WWAV), the GP1 of Tamiami virus (TMMV), the GP1 of Bear Canyon virus (BCNV), the GP1 of Skinner Tank virus (SKTV), and the GP1 of Catarina virus (CTNV).

[0179] In some embodiments, the antibody or antigen binding portion thereof disclosed herein binds to the GP1 of Junin virus (JUNV). In some embodiments, the antibodies disclosed herein, or antigen binding portions thereof, bind to the GP1 of Machupo virus (MACV). In some embodiments, the antibodies disclosed herein, or antigen binding portions thereof, bind to the GP 1 of Sabia virus (SBAV). In some embodiments, the antibodies disclosed herein, or antigen binding portions thereof, bind to the GP1 of SBAV-like virus (SBAV-like). In some embodiments, the antibodies disclosed herein, or antigen binding portions thereof, bind to the GP1 of Chapare virus (CHAPV).

[0180] In some embodiments, the antibody or antigen binding portion thereof disclosed herein binds to the GP1 of JUNV and the GP1 of MACV.

[0181] In some embodiments, the antibody or antigen binding portion thereof disclosed herein binds to GP1 of one or more arenaviruses, wherein the GP1 is selected from the group consisting of the GP1 of JUNV, the GP1 of MACV, the GP1 of SBAV, the GP1 of SBAV-like, and the GP1 of CHAPV.

[0182] GP1 subunit'. Arenaviruses exist worldwide and can cause hemorrhagic fever and neurologic diseases. A glycoprotein is expressed on the viral surface that mediates entry into target cells. This glycoprotein, termed GPC, contains a membrane-associated signal peptide, a receptor-binding subunit termed GP1 and a fusion-mediating subunit termed GP2. After receptor binding, arenaviruses enter via endocytosis. Exposure of their GPC to acidic pH in the target cell endosome triggers dissociation of GP1 from GP2, and irreversible conformational changes in GP2 which drive fusion of virus and host membranes. As used herein, unless stated otherwise, GP1 includes any arenavirus form of GP1.

[0183] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 33, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 33.Atorney Docket No. 117823-37620

[0184] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 34, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 34.

[0185] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 35, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 35.

[0186] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 36, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 36.

[0187] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 37, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 37.

[0188] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 38, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 38.

[0189] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 39, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 39.

[0190] The GP1 of arenavirus may, for example, comprise the amino acid sequence of SEQ ID NO: 40, or any portions, mutants, splice variants, isoforms, orthologs, homologs, and variants of this sequence. In some embodiments, the GP1 comprises a polypeptide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 40.

[0191] In some instances, an anti-GPl antibody or antigen binding fragment thereof according to the disclosure binds to a GP1 of an arenavirus (e.g., the GP1 of JUNV, the GP1 of MACV, the GP1 of SBAV, the GPl of SBAV-like, and / orthe GPl of CHAPV, and / or any of the GP1 proteins listed herein) with a dissociation constant (KD) of 100 nM or lower; about 10 nM or lower; about 1 nM or lower; about 100 pM or lower; about 10 pM or lower; about 1 pM or lower; or about 0.1 pM or lower.Atorney Docket No. 117823-37620

[0192] The present disclosure provides exemplary antibodies or antigen binding fragments thereof that bind a GP1 of an arenavirus, which optionally may be affinity-matured. Other antibodies or antigen binding fragments thereof that bind a GP1 of an arenavirus, including those having different CDRs, and epitopic specificity may be obtained using the disclosure of the present specification, and using methods that are generally known in the art. Such antibodies and antigen binding fragments thereof antagonize the biological effects of a GP 1 of an arenavirus in vivo and therefore are useful in treating or preventing arenavirus-related conditions including, particularly arenavirus infection. In preferred embodiments, the antibody or antigen binding fragment thereof according to the disclosure comprises one or more CDRs, a VL chain and / or VH chain of the anti-GPl antibodies and antigen binding fragments thereof described herein.

[0193] In some embodiments, an anti-GP 1 antibody or antigen binding fragment thereof according to the disclosure can interfere with, block, reduce, or modulate the interaction between GP1 and its receptor (e.g., transferrin receptor 1 (TfRl)) on host cells. If binding of the GP1 protein to its receptor is blocked or reduced, an arenavirus may be prohibited from entering the cells, thereby preventing infection to further cells.

[0194] In some examples, an anti-GPl antibody or antigen binding fragment thereof according to the disclosure is “neutralizing”, e.g., it substantially or totally prevents the specific interaction of arenavirus with the host receptors or priming protein. As a result, virions may be substantially or totally cleared by immune cells of the host, such as phagocytes via, for example, Fc receptor mediated phagocytosis or mere phagocytosis due to increased time of virions outside the cells. In some embodiments, the antibody or antigen binding fragment thereof neutralizes arenavirus, e.g., by remaining bound to arenavirus in a location and / or manner that prevents arenavirus from binding to its receptor or priming protein on host cells. As a result, arenavirus virions may be substantially or totally prevented from entering the cells, thereby preventing infection to further cells.

[0195] In certain embodiments, an anti-GPl antibody or antigen binding fragment thereof according to the disclosure neutralizes arenavirus (e.g., JUNV, SBAV, SBAV-like, MACV, and / or CHAPV) at an IC50 of about 100 nM or lower, of about 50 nM or lower, of about 20 nM or lower, of about 10 nM or lower, of about 5 nM or lower, of about 2 nM or lower, of about 1 nM or lower, of about 500 pM or lower, of about 200 pM or lower, of about 100 pM or lower, of about 50 pM or lower, of about 20 pM or lower, of about 10 pM or lower, of about 5 pM or lower, of about 2 pM or lower, or of about 1 pM or lower, or at an IC50 of about 500 ng / mL or lower, of about 400 ng / mL or lower, of about 300 ng / mL or lower, of about 200 ng / mL or lower, of about 150 ng / mL or lower, of about 100 ng / mL or lower, of about 90 ng / mL or lower, of about 80 ng / mL or lower, of about 70 ng / mL or lower, of about 60 ng / mL or lower, of about 50 ng / mL or lower, of about 40 ng / mL or lower, of about 30 ng / mL or lower, of about 20 ng / mL or lower, of about 10 ng / mL or lower, of about 5 ng / mL or lower, of about 2 ng / mL or lower, or at about 1 ng / mL or lower, in vitro.Atorney Docket No. 117823-37620

[0196] In some embodiments, the anti-GP 1 antibody or antigen binding fragment thereof according to the disclosure neutralizes arenavirus (e.g., JUNV, SBAV, SBAV-like, MACV, and / or CHAPV) at an IC50 of about 100 ng / mL or lower, in vitro.

[0197] In some embodiments, the anti-GP 1 antibody or antigen binding fragment thereof according to the disclosure neutralizes arenavirus (e.g., JUNV, SBAV, SBAV-like, MACV, and / or CHAPV) at an IC50 of about 50 ng / mL or lower, in vitro.

[0198] In some embodiments, the anti-GP 1 antibody or antigen binding fragment thereof according to the disclosure neutralizes JUNV at an IC50 of about 100 ng / mL or lower, in vitro.

[0199] In some embodiments, the anti-GP 1 antibody or antigen binding fragment thereof according to the disclosure neutralizes JUNV at an IC50 of about 50 ng / mL or lower, in vitro.

[0200] While the term “antibody” is used throughout, it should be noted that antibody fragments (i.e., antigen binding portions of an anti-GP 1 antibody) are also included in the disclosure and may be included in the embodiments (methods and compositions) described throughout. For example, an anti-GP 1 antibody fragment may be conjugated to the drugs, as described herein. In certain embodiments, an anti-GP 1 antibody binding portion is a Fab, a Fab’, a F(ab’)2, a Fv, a disulfide linked Fv, an scFv, a single domain antibody, or a diabody.

[0201] In some embodiments, the heavy and light chain variable region amino acid sequences for the antibodies are set forth in Table 2.

[0202] Thus, in one embodiment, the disclosure includes anti-GP 1 antibodies, or antigen binding portions thereof, comprising a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 9, 17, and 25; and a light chain variable region comprising an amino acid sequence selected from the group consisting of 5, 13, 21, and 29.

[0203] In one embodiment, the disclosure includes an anti-GP 1 antibody, or antigen binding portion thereof, comprising an heavy chain CDR set (CDR1, CDR2, and CDR3) selected from those set forth in Table 2; and an light chain CDR set (CDR1, CDR2, and CDR3) selected from those set forth in Table 2.

[0204] In one embodiment, an anti-GP 1 antibody, or antigen binding portion thereof, is the human antibody AHF1 B7. In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 4 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 8.

[0205] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 3, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 2, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 8, a CDR2 domain comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequenceAtorney Docket No. 117823-37620

[0206] of SEQ ID NO: 6. In further embodiments, disclosed herein is an antibody having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5.

[0207] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 1, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 5, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 5.

[0208] In one embodiment, the disclosure features an anti-GPl antibody, or antigen binding portion thereof, which is the human antibody AHF2 C5. In some embodiments, an anti-GP 1 antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 12 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 16.

[0209] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 11, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 domain comprising the amino acid sequence EVS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 14. In further embodiments, the disclosure provides an antibody having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 13.

[0210] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 9, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 9, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 13, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13.

[0211] In one embodiment, the disclosure features an anti-GPl antibody, or antigen binding portion thereof, which is the human antibody AHF2 C8. In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 20 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 24.

[0212] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 20, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 19, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 18, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 24, a CDR2 domainAtorney Docket No. 117823-37620

[0213] comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 22. In further embodiments, the disclosure provides an antibody having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 21.

[0214] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21.

[0215] In one embodiment, the disclosure features an anti-GPl antibody, or antigen binding portion thereof, which is the human antibody AHF3 E8.2. In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 28 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 32 .

[0216] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 28, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 27, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 26, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 32, a CDR2 domain comprising the amino acid sequence DAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 30. In further embodiments, the disclosure provides an antibody having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 29.

[0217] In some embodiments, an anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 29.

[0218] The foregoing anti-GPl antibody CDR sequences establish a novel family of GP1 binding proteins, isolated in accordance with this disclosure, and comprising antigen binding polypeptides that include the CDR sequences listed in Table 2, as well as the Sequence Summary.

[0219] To generate and to select CDRs having preferred GP1 binding and / or neutralizing activity with respect to GP1, standard methods known in the art for generating antibodies, or antigen binding portions thereof, and assessing the GP1 binding and / or neutralizing characteristics of those antibodies, or antigen binding portions thereof, may be used, including but not limited to those specifically described herein.Atorney Docket No. 117823-37620

[0220] In certain embodiments, the antibody comprises a heavy chain constant region, such as an IgGl, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant region. In certain embodiments, the anti-GP1 antibody, or antigen binding portion thereof, comprises a heavy chain immunoglobulin constant domain selected from the group consisting of a human IgG constant domain, a human IgM constant domain, a human IgE constant domain, and a human IgA constant domain. In further embodiments, the antibody, or antigen binding portion thereof, has an IgGl heavy chain constant region, an IgG2 heavy chain constant region, an IgG3 constant region, or an IgG4 heavy chain constant region.

[0221] Preferably, the heavy chain constant region is an IgGl heavy chain constant region or an IgG4 heavy chain constant region. In one embodiment, the antibody, or antigen binding portion thereof, is an IgG4 isotype.

[0222] Furthermore, the antibody can comprise a light chain constant region, either a kappa light chain constant region or a lambda light chain constant region. Preferably, the antibody comprises a kappa light chain constant region. Alternatively, the antibody portion can be, for example, a Fab fragment or a single chain Fv fragment.

[0223] In certain embodiments, the anti-GPl antibody binding portion is a Fab, a Fab’, a F(ab’)2, a Fv, a disulfide linked Fv, an scFv, a single domain antibody, or a diabody.

[0224] In certain embodiments, the anti-GP 1 antibody, or antigen binding portion thereof, is a multispecific antibody, e.g. a bispecific antibody.

[0225] In certain embodiments, the anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 1 and / or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 5.

[0226] In certain embodiments, the anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 9 and / or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 13.

[0227] In certain embodiments, the anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 17 and / or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 21.

[0228] In certain embodiments, the anti-GPl antibody, or antigen binding portion thereof, comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 25 and / or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 29.

[0229] Replacements of amino acid residues in the Fc portion to alter antibody effector function are have been described (Winter, et al. US Patent Nos. 5,648,260 and 5,624,821, incorporated by reference herein). The Fc portion of an antibody mediates several important effector functions e.g. cytokine induction, ADCC, phagocytosis, complement dependent cytotoxicity (CDC) and half-life / clearance rate of antibody and antigen-antibody complexes. In some cases these effector functions are desirable for therapeutic antibody but in other cases might be unnecessary or even deleterious, depending on the therapeutic objectives. Certain human IgG isotypes, particularly IgGl and IgG3,Atorney Docket No. 117823-37620

[0230] mediate ADCC and CDC via binding to FcyRs and complement Clq, respectively. Neonatal Fc receptors (FcRn) are the critical components determining the circulating half-life of antibodies. In still another embodiment at least one amino acid residue is replaced in the constant region of the antibody, for example the Fc region of the antibody, such that effector functions of the antibody are altered.

[0231] One embodiment includes a labeled anti-GPl antibody, or antibody portion thereof, where the antibody is derivatized or linked to one or more functional molecule(s) (e.g., another peptide or protein). For example, a labeled antibody can be derived by functionally linking an antibody or antibody portion of the disclosure (by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or a diabody), a detectable agent, a pharmaceutical agent, a protein or peptide that can mediate the association of the antibody or antibody portion with another molecule (such as a streptavidin core region or a polyhistidine tag), and / or a cytotoxic or therapeutic agent selected from the group consisting of a mitotic inhibitor, an antitumor antibiotic, an immunomodulating agent, a vector for gene therapy, an alkylating agent, an antiangiogenic agent, an antimetabolite, a boron-containing agent, a chemoprotective agent, a hormone, an antihormone agent, a corticosteroid, a photoactive therapeutic agent, an oligonucleotide, a radionuclide agent, a topoisomerase inhibitor, a tyrosine kinase inhibitor, a radiosensitizer, and a combination thereof.

[0232] Useful detectable agents with which an antibody or antibody portion thereof, may be derivatized include fluorescent compounds. Exemplary fluorescent detectable agents include fluorescein, fluorescein isothiocyanate, rhodamine, 5 -dimethylamine- 1-napthalenesulfonyl chloride, phycoerythrin and the like. An antibody may also be derivatized with detectable enzymes, such as alkaline phosphatase, horseradish peroxidase, glucose oxidase and the like. When an antibody is derivatized with a detectable enzyme, it is detected by adding additional reagents that the enzyme uses to produce a detectable reaction product. For example, when the detectable agent horseradish peroxidase is present the addition of hydrogen peroxide and diaminobenzidine leads to a colored reaction product, which is detectable. An antibody may also be derivatized with biotin, and detected through indirect measurement of avidin or streptavidin binding.

[0233] In one embodiment, the antibody is conjugated to an imaging agent. Examples of imaging agents that may be used in the compositions and methods described herein include, but are not limited to, a radiolabel (e.g., indium), an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.

[0234] In one embodiment, the antibody or antibody portion thereof disclosed herein is linked to a radiolabel, such as, but not limited to, indium (”’In).11’Indium may be used to label the antibody or antibody portion thereof disclosed herein for use in identifying GP1 positive cells. In a certain embodiment, the antibody or antibody portion thereof disclosed herein is labeled with11’I via a bifunctional chelator which is a bifunctional cyclohexyl diethylenetriaminepentaacetic acid (DTP A)Atorney Docket No. 117823-37620

[0235] chelate (see US Patent Nos. 5,124,471; 5,434,287; and 5,286,850, each of which is incorporated herein by reference).

[0236] Another embodiment of the disclosure provides a glycosylated binding protein wherein the anti-GPl antibody or antigen binding portion thereof comprises one or more carbohydrate residues. Nascent in vivo protein production may undergo further processing, known as post-translational modification. In particular, sugar (glycosyl) residues may be added enzymatically, a process known as glycosylation. The resulting proteins bearing covalently linked oligosaccharide side chains are known as glycosylated proteins or glycoproteins. Antibodies are glycoproteins with one or more carbohydrate residues in the Fc domain, as well as the variable domain. Carbohydrate residues in the Fc domain have important effect on the effector function of the Fc domain, with minimal effect on antigen binding or half-life of the antibody (R. Jefferis, Biotechnol. Prog. 21 (2005), pp. 11-16). In contrast, glycosylation of the variable domain may have an effect on the antigen binding activity of the antibody. Glycosylation in the variable domain may have a negative effect on antibody binding affinity, likely due to steric hindrance (Co, M.S., et al., Mol. Immunol. (1993) 30: 1361- 1367), or result in increased affinity for the antigen (Wallick, S.C., et al. , Exp. Med. (1988) 168: 1099-1109; Wright, N., etal., EMBO J. (1991) 10:2717-2723).

[0237] One aspect of the disclosure is directed to generating glycosylation site mutants in which the O- or N-linked glycosylation site of the binding protein has been mutated. One skilled in the art can generate such mutants using standard well-known technologies. Glycosylation site mutants that retain the biological activity, but have increased or decreased binding activity, are another object of the disclosure.

[0238] In still another embodiment, the glycosylation of the anti-GP 1 antibody or antigen binding portion is modified. For example, an aglycoslated antibody can be made (i.e., the antibody lacks glycosylation). Glycosylation can be altered to, for example, increase the affinity of the antibody for antigen. Such carbohydrate modifications can be accomplished by, for example, 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 elimination of one or more variable region glycosylation sites to thereby eliminate glycosylation at that site. Such aglycosylation may increase the affinity of the antibody for antigen. Such an approach is described in further detail in PCT Publication W02003016466A2, and U.S. Pat. Nos. 5,714,350 and 6,350,861, each of which is incorporated herein by reference in its entirety.

[0239] Additionally or alternatively, a modified anti-GP 1 antibody can be made that has an altered type of glycosylation, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNAc structures. Such altered glycosylation paterns have been demonstrated to increase the ADCC ability of antibodies. Such carbohydrate modifications can be accomplished by, for example, expressing the antibody in a host cell with altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be usedAtorney Docket No. 117823-37620

[0240] as host cells in which to express recombinant antibodies to thereby produce an antibody with altered glycosylation. See, for example, Shields, R. L. etal. (2002) J. Biol. Chem. 277:26733-26740; Umana et al. (1999) Nat. Biotech. 17:176-1, as well as, European Patent No: EP 1,176,195; PCT Publications WO 03 / 035835; WO 99 / 54342 80, each of which is incorporated herein by reference in its entirety.

[0241] Protein glycosylation depends on the amino acid sequence of the protein of interest, as well as the host cell in which the protein is expressed. Different organisms may produce different glycosylation enzymes (e.g., glycosyltransferases and glycosidases), and have different substrates (nucleotide sugars) available. Due to such factors, protein glycosylation patern, and composition of glycosyl residues, may differ depending on the host system in which the particular protein is expressed. Glycosyl residues useful may include, but are not limited to, glucose, galactose, mannose, fucose, n-acetylglucosamine and sialic acid. Preferably the glycosylated binding protein comprises glycosyl residues such that the glycosylation patern is human.

[0242] Differing protein glycosylation may result in differing protein characteristics. For instance, the efficacy of a therapeutic protein produced in a microorganism host, such as yeast, and glycosylated utilizing the yeast endogenous pathway may be reduced compared to that of the same protein expressed in a mammalian cell, such as a CHO cell line. Such glycoproteins may also be immunogenic in humans and show reduced half-life in vivo after administration. Specific receptors in humans and other animals may recognize specific glycosyl residues and promote the rapid clearance of the protein from the bloodstream. Other adverse effects may include changes in protein folding, solubility, susceptibility to proteases, trafficking, transport, compartmentalization, secretion, recognition by other proteins or factors, antigenicity, or allergenicity. Accordingly, a practitioner may prefer a therapeutic protein with a specific composition and patern of glycosylation, for example glycosylation composition and patern identical, or at least similar, to that produced in human cells or in the species-specific cells of the intended subject animal.

[0243] Expressing glycosylated proteins different from that of a host cell may be achieved by genetically modifying the host cell to express heterologous glycosylation enzymes. Using recombinant techniques, a practitioner may generate antibodies or antigen binding portions thereof exhibiting human protein glycosylation. For example, yeast strains have been genetically modified to express non-naturally occurring glycosylation enzymes such that glycosylated proteins (glycoproteins) produced in these yeast strains exhibit protein glycosylation identical to that of animal cells, especially human cells (U.S. patent Publication Nos. 20040018590 and 20020137134 and PCT publication W02005100584 A2).

[0244] Antibodies may be produced by any of a number of techniques. For example, expression from host cells, wherein expression vector(s) encoding the heavy and light chains is (are) transfected into a host cell by standard techniques. The various forms of the term “transfection” are intended to encompass a wide variety of techniques commonly used for the introduction of exogenous DNA into a prokaryotic or eukaryotic host cell, e.g., electroporation, calcium-phosphate precipitation, DEAE-Atorney Docket No. 117823-37620

[0245] dextran transfection and the like. Although it is possible to express antibodies in either prokaryotic or eukaryotic host cells, expression of antibodies in eukaryotic cells is preferable, and most preferable in mammalian host cells, because such eukaryotic cells (and in particular mammalian cells) are more likely than prokaryotic cells to assemble and secrete a properly folded and immunologically active antibody.

[0246] Preferred mammalian host cells for expressing the recombinant antibodies disclosed herein include Chinese Hamster Ovary (CHO cells) (including dhfir- CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, used with a DHFR selectable marker, e.g., as described in R.J. Kaufman and P.A. Sharp (1982) Mol. Biol. 159:601-621), NS0 myeloma cells, COS cells and SP2 cells. When recombinant expression vectors encoding antibody genes are introduced into mammalian host cells, the antibodies are produced by culturing the host cells for a period of time sufficient to allow for expression of the antibody in the host cells or, more preferably, secretion of the antibody into the culture medium in which the host cells are grown. Antibodies can be recovered from the culture medium using standard protein purification methods.

[0247] Host cells can also be used to produce functional antibody fragments, such as Fab fragments or scFv molecules. It will be understood that variations on the above procedure are within the scope of the disclosure. For example, it may be desirable to transfect a host cell with DNA encoding functional fragments of either the light chain and / or the heavy chain of an antibody. Recombinant DNA technology may also be used to remove some, or all, of the DNA encoding either or both of the light and heavy chains that is not necessary for binding to the antigens of interest. The molecules expressed from such truncated DNA molecules are also encompassed by the antibodies of the disclosure. In addition, bifunctional antibodies may be produced in which one heavy and one light chain are an antibody of the disclosure and the other heavy and light chain are specific for an antigen other than the antigens of interest by crosslinking an antibody of the disclosure to a second antibody by standard chemical crosslinking methods.

[0248] In a preferred system for recombinant expression of an antibody, or antigen binding portion thereof, a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is introduced into CHO cells comprising a glutamine synthase expression system, commercially available from Lonza (hereafter GS-CHO) (Bebbington, C. R. et al. (1992), Biotechnology, 10, pages 169-175).

[0249] In another system for recombinant expression of an antibody, or antigen binding portion thereof, a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is introduced into dhfr- CHO cells by calcium phosphate-mediated transfection. Within the recombinant expression vector, the antibody heavy and light chain genes are each operatively linked to CMV enhancer / AdMLP promoter regulatory elements to drive high levels of transcription of the genes. The recombinant expression vector also carries a DHFR gene, which allows for selection of CHO cells that have been transfected with the vector using methotrexate selection / amplification.Atorney Docket No. 117823-37620

[0250] The selected transformant host cells are cultured to allow for expression of the antibody heavy and light chains and intact antibody is recovered from the culture medium. Standard molecular biology techniques are used to prepare the recombinant expression vector, transfect the host cells, select for transformants, culture the host cells and recover the antibody from the culture medium. Still further the disclosure provides a method of synthesizing a recombinant antibody by culturing a host cell in a suitable culture medium until a recombinant antibody is synthesized. Recombinant antibodies may be produced using nucleic acid molecules corresponding to the amino acid sequences disclosed herein. The method can further comprise isolating the recombinant antibody from the culture medium.

[0251] III. Cross-Reactive Anti-GPl Antibodies

[0252] The present invention relates to anti-GPl antibodies, and antigen binding portions thereof, which cross-react with two or more arenaviruses. In some embodiments, the present disclosure provides exemplary human anti-GPl antibodies, including, for example, AHF1 B7, AHF2 C5, AHF2 C8, and AHF3 E8.2, their antigen binding portions, including but not limited to their complementarity determining regions (CDRs), derivatives, and variants thereof, which display broad and potent neutralization for arenaviruses in vitro and / or in vivo. Thus, the anti-GPl antibodies of the present invention possess strong therapeutic utility, either alone or in combination with one another, or with other neutralizing antibodies for arenaviruses, or with other arenavirus treatments, as discussed herein.

[0253] A “cross-reactive antibody” or “broadly neutralizing antibody” is an antibody that binds to and inhibits the function of related antigens (e.g., GP1), such as antigens that share at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity and / or a similar motif at the area in contact with the antibody. With regard to an antigen from a pathogen, such as a virus, the antibody can bind to and inhibit the function of an antigen from more than one class and / or subclass of the pathogen. For example, with regard to an arenavirus, the antibody can bind to and inhibit the function of an antigen, such as GP1 from more than one clade.

[0254] The data in the Example section herein show that heavy chain variable domain (VH) of the assessed antibody confers the majority of the surface contact area between the antibody and GP1 protein. The exemplary data disclosed herein further show that the antibodies contain a CX4-5CY motif, in which the paired cysteine residues can form disulfide bridges that support the extended length of the CDRH3, allowing for flexible but stabilized insertion into the GP1 pocket. Notably, the CDRH3 is responsible for almost all of the heavy chain contact by contributing more than about 85% of the total VH buried surface area. Thus, antibodies comprising the CX4-5CY motif can be candidates for broadly neutralizing antibodies for arenaviruses.

[0255] In some aspects of the invention, the present disclosure provides an isolated antibody, or antigen binding fragment thereof, which binds the same epitope as the isolated antibodies, or antigen binding fragments thereof, disclosed herein. In some embodiments, the isolated antibody or antigenAtorney Docket No. 117823-37620

[0256] binding fragment thereof disclosed herein cross-reacts with two or more arenaviruses (e.g., three or more arenaviruses, four or more arenaviruses, five or more arenaviruses, or six or more arenaviruses). In some embodiments, the heavy chain variable region comprises a CDR3 that comprises a CXXXXCY motif or a CXXXXXCY motif, wherein X is any amino acid residue.

[0257] In some embodiments, the two or more arenaviruses are selected from the group consisting of JUNV, MACV, SBAV, SBAV-like, and CHAPV.

[0258] In some embodiments, the epitope of the GP1 comprises an asparagine (D), a serine (S), and a positive charged amino acid residue. In some embodiments, the positive charged amino acid residue is selected from group consisting of lysine (K) and arginine (R).

[0259] In some embodiments, the GP1 comprises an epitope comprising a serine (S), an asparagine (D), an isoleucine (I), a valine (V), a tyrosine (Y), an lysine (K), and a glutamic acid (E).

[0260] In some embodiments, the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or all, of amino acid residues Sill, DI 13, DI 14, 1115, Al 16, VI 17, LI 19, Y122, E171, K216, and Q218, or any combination thereof of the amino acid sequence set forth in SEQ ID NO: 33.

[0261] In some embodiments, the GP1 comprises an epitope comprising amino acid residues Sill, DI 13, DI 14, 1115, Al 16, VI 17, LI 19, Y122, K169, E171, K216, and Q218, or any combination thereof of the amino acid sequence set forth in SEQ ID NO: 33.

[0262] In some embodiments, the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all, amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 19, Y122, K169, E171, K216, and Q218, or any combination thereof of the amino acid sequence set forth in SEQ ID NO: 33.

[0263] In some embodiments, the GP1 comprises an epitope comprising amino acid residues Sill, D113, D114, 1115, A116, V117, L118, L119, E121, Y122, D123, R165, E171, K216, and Q218, or any combination thereof of the amino acid sequence set forth in SEQ ID NO: 33.

[0264] In some embodiments, the GP1 comprises an epitope comprising at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or all, amino acid residues S97, F98, Rill, VI 12, SI 13, DI 14, Il 15, SI 16, VI 17, Ml 19, Y122, R165, K169, K170, E171, N174, T216, N218, F226, and Y228, or any combination thereof of thean amino acid sequence set forth in SEQ ID NO: 35.

[0265] Amino acid residues can be found in Table 1.

[0266] Table 1. Amino Acid Residues.

[0267]

[0268] Atorney Docket No. 117823-37620

[0269]

[0270] Atorney Docket No. 117823-37620

[0271] In some embodiments, the heavy chain variable region comprises a CDR3 that contributes at least about 70% (e.g., at least about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%) of the contact area between the isolated antibody, or antigen binding portion thereof, and the GP1. In some embodiments, the heavy chain variable region comprises a CDR3 that contributes at least about 75% of the contact area between the isolated antibody, or antigen binding portion thereof, and the GP 1. In some embodiments, the heavy chain variable region comprises a CDR3 that contributes at least about 80% of the contact area between the isolated antibody, or antigen binding portion thereof, and the GP1. In some embodiments, the heavy chain variable region comprises a CDR3 that contributes at least about 82% of the contact area between the isolated antibody, or antigen binding portion thereof, and the GP1.

[0272] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 4.

[0273] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 12.

[0274] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 20.

[0275] In some embodiments, the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 28.

[0276] IV. Uses of Anti-GPl Antibodies

[0277] The present disclosure provides methods for ameliorating or reducing the symptoms of, or treating, or preventing, diseases and disorders associated with an arenavirus. The methods comprise administering an antibody, or antigen binding fragment thereof, that displays broad activity against one or more arenaviruses.

[0278] In some aspects, disclosed herein is a method of treating or preventing an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with the infection in a subject in need thereof. The method includes administering to the subject a therapeutically effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, thereby treating or preventing an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with the infection in the subject.

[0279] In some aspects, disclosed herein is a method of inducing an immune response against one or more arenaviruses in a subject in need thereof. The method includes administering to the subject anAtorney Docket No. 117823-37620

[0280] effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, thereby inducing an immune response against one or more arenaviruses in the subject.

[0281] In some embodiments, the immune response elicits immunoprotection against the one or more arenaviruses.

[0282] In some aspects, disclosed herein is a method of inhibiting or blocking infection of susceptible cells by one or more arenaviruses in a subject in need thereof. The method includes administering to the subject an effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, thereby inhibiting or blocking infection of susceptible cells by one or more arenaviruses in the subject.

[0283] In some aspects, disclosed herein is a method of preventing the onset of a hemorrhagic fever in a subject infected by one or more arenaviruses, or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in a subject infected by one or more arenaviruses. The method includes administering to the subject a prophylactically or therapeutically effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, thereby preventing the onset of a hemorrhagic fever or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in the subject, antigen binding

[0284] In some embodiments, the arenavirus is an Old World arenavirus or a New World arenavirus. In some embodiments, the one or more arenaviruses are selected from the group consisting of Lymphocytic choriomeningitis virus (LCMV), Lassa virus (LASV), Lujo virus, Kodoko virus (KODV), Mobala virus (MOBV), Ippy virus (IPPYV), Mopeia virus (MOPV), Morogoro virus (MORV), Allpahuayo virus (AALV), Flexal virus (FLEV), Parana virus (PRAV), Pichinde virus (PICHV), Pirital virus (PIRV), Chapare virus (CHAPV), Sabia virus (SBAV), Cupixi virus (CPXV), Guanarito virus (GTOV), Amapari virus (AMAV), Junin virus (JUNV), Machupo virus (MACV), Tacaribe virus (TCRV), Oliveros virus (OLVV), Latino virus (LATV), Whitewater Arroyo virus (WWAV), Tamiami virus (TMMV), Bear Canyon virus (BCNV), Skinner Tank virus (SKTV), and Catarina virus (CTNV).

[0285] In some embodiments, the one or more arenaviruses are selected from the group consisting of Junin virus (JUNV), Machupo virus (MACV), Sabia virus (SBAV), SBAV-like virus (SBAV-like), and Chapare virus (CHAPV).

[0286] Example 1 shows that antibody AHF1 B7 and antibody AHF2 C5 show activities against JUNV, MACV, SBAV, and SBAV-like; antibody AHF2_C8 shows activities against JUNV, SBAV, SBAV-like, and CHAPV; and antibody AHF3_E8.2 shows activities against JUNV, MACV, SBAV, SBAV-like, and CHAPV.

[0287] Accordingly, in some aspects, disclosed herein is a method of treating or preventing an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with the infection in a subject in need thereof, inhibiting or blocking infection ofAtorney Docket No. 117823-37620

[0288] susceptible cells by one or more arenaviruses in a subject in need thereof, preventing the onset of a hemorrhagic fever in a subject infected by one or more arenaviruses, or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in a subject infected by one or more arenaviruses. The method includes administering to the subject an effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, wherein the one or more arenaviruses are selected from the group consisting of JUNV, MACV, SB AV, and SBAV-like, and wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 3, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 2, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 8, a CDR2 domain comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 6.

[0289] In further embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5. In some embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, comprising a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 1, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 5, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 5.

[0290] In some aspects, disclosed herein is a method of treating or preventing an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with the infection in a subject in need thereof, inhibiting or blocking infection of susceptible cells by one or more arenaviruses in a subject in need thereof, preventing the onset of a hemorrhagic fever in a subject infected by one or more arenaviruses, or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in a subject infected by one or more arenaviruses. The method includes administering to the subject an effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, wherein the one or more arenaviruses are selected from the group consisting of JUNV, MACV, SBAV, and SBAV-like, and wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 11, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 domain comprising the amino acid sequence EVS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 14.Atorney Docket No. 117823-37620

[0291] In further embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, comprising a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 9, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 9, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 13, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13.

[0292] In some aspects, disclosed herein is a method of treating or preventing an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with the infection in a subject in need thereof, inhibiting or blocking infection of susceptible cells by one or more arenaviruses in a subject in need thereof, preventing the onset of a hemorrhagic fever in a subject infected by one or more arenaviruses, or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in a subject infected by one or more arenaviruses. The method includes administering to the subject an effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, wherein the one or more arenaviruses are selected from the group consisting of JUNV, SBAV, SBAV-like, and CHAPV, and wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 20, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 19, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 18, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 24, a CDR2 domain comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 22.

[0293] In further embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 21. In some embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, comprising a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21.

[0294] In some aspects, disclosed herein is a method of treating or preventing an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with the infection in a subject in need thereof, inhibiting or blocking infection of susceptible cells by one or more arenaviruses in a subject in need thereof, preventing the onset of a hemorrhagic fever in a subjectAtorney Docket No. 117823-37620

[0295] infected by one or more arenaviruses, or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in a subject infected by one or more arenaviruses. The method includes administering to the subject an effective amount of an antibody, or antigen binding fragment thereof, disclosed herein, or a pharmaceutical composition disclosed herein, wherein the one or more arenaviruses are selected from the group consisting of JUNV, MACV, SBAV, SBAV-like, and CHAPV, and wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 28, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 27, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 26, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 32, a CDR2 domain comprising the amino acid sequence DAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 30.

[0296] In further embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 29. In some embodiments, the method includes administering to the subject an effective amount of an antibody, or antigen binding portion thereof, comprising a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 29.

[0297] Symptoms of arenavirus infection may include fever, malaise, headache, conjunctivitis, nausea, vomiting, diarrhea, thrombocytopenia, bleeding, neurological disorders, petechiae (such as petechiae in oral mucosa, chest, arms, or axillary region), ataxia, tremors, seizures, and coma.

[0298] Arenavirus infection may cause hemorrhagic fever. Viral hemorrhagic fevers are infectious diseases that can cause severe, life-threatening illness. They can damage the walls of tiny blood vessels, making them leak, and can hamper the blood's ability to clot. Early signs and symptoms of hemorrhagic fever can include, for example, fever, fatigue, weakness, dizziness, muscle, bone, or, joint aches, nausea and vomiting, or diarrhea. More-severe symptoms of hemorrhagic fever can include, for example, bleeding under the skin, in internal organs, or from the mouth, eyes or ears, nervous system malfunctions, coma, delirium, kidney failure, respiratory failure, or liver failure. In some embodiments, the hemorrhagic fever is selected from the group consisting of Argentine hemorrhagic fever, Bolivian hemorrhagic fever, Brazilian hemorrhagic fever, and hemorrhagic fever caused by CHAPV.

[0299] The subject to which the pharmaceutical formulation is administered can be, e.g., any human or non-human animal needing such treatment, prevention and / or amelioration, or who would otherwise benefit from the inhibition or atenuation of arenavirus-mediated activity. For example, theAtorney Docket No. 117823-37620

[0300] subject can be an individual that is diagnosed with, or who is deemed to be at risk of being afflicted by any of the aforementioned diseases or disorders. In some instances the subject may be in an advanced state of arenavirus infection, e.g., a subject who has hemorrhagic fever or neurological disorders. In some instances, the subject can be one having one or more risk factors which correlate to a poor arenavirus treatment or recovery prognosis. The present disclosure further includes the use of any of the pharmaceutical formulations disclosed herein in the manufacture of a medicament for the treatment, prevention and / or amelioration of any disease or disorder associated with arenavirus activity (including any of the above-mentioned exemplary diseases, disorders and conditions).

[0301] In another aspect, this application features a method of treating (e.g., curing, suppressing, ameliorating, delaying or preventing the onset of, or preventing recurrence or relapse of) or preventing an arenavirus-associated disorder, in a subject. The method includes: administering to the subject an anti-GPl antibody or fragment thereof as described herein, in an amount sufficient to treat or prevent the arenavirus-associated disorder. The anti-GP 1 antibody or fragment thereof, can be administered to the subject, alone or in combination with other therapeutic modalities as described herein.

[0302] In some embodiments, the anti-GP 1 antibody or fragment thereof used in the methods of the invention is a human or humanized anti-GP 1 antibody or fragment thereof.

[0303] In another embodiment, antibody-dependent cell-mediated cytotoxicity (ADCC) activity is not necessary for anti-GPl antibodies to inhibit arenavirus replication or reduce arenavirus levels. Accordingly, in one embodiment, an antibody, or antigen binding portion thereof, of the invention comprises an isotype lacking effector function (e.g., human IgG4).

[0304] Antibodies, or antigen binding portions thereof, can be used alone or in combination to treat such diseases. It should be understood that the antibodies or antigen binding portion thereof can be used alone or in combination with an additional agent, e.g., a therapeutic agent, said additional agent being selected by the skilled artisan for its intended purpose. For example, the additional agent can be a therapeutic agent art-recognized as being useful to treat the disease or condition being treated by the antibody. The additional agent also can be an agent that imparts a beneficial atribute to the therapeutic composition, e.g., an agent which affects the viscosity of the composition.

[0305] It should further be understood that the combinations which are to be included within this disclosure are those combinations useful fortheir intended purpose. The agents set forth below are illustrative for purposes and not intended to be limited. The combinations, which are part of this disclosure, can be the antibodies of the disclosure and at least one additional agent selected from the lists below. The combination can also include more than one additional agent, e.g., two or three additional agents if the combination is such that the formed composition can perform its intended function.

[0306] Dosage regimens may be adjusted to provide the optimum desired response (e.g., a therapeutic or prophylactic response). For example, a single bolus may be administered, several divided doses mayAtorney Docket No. 117823-37620

[0307] be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.

[0308] In some embodiments, the antibodies and fragments thereof described herein or the pharmaceutical composition are administered to a subject at a dose of between about 0.01 and 100.0 or 200.0 mg / kg of body weight of the recipient subject. In certain embodiments, depending on the type and severity of the arenavirus-related disease, about 1 pg / kg to 50 mg / kg (e.g., 0.1-20 mg / kg) of antibody is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. In another embodiment, about 1 pg / kg to 15 mg / kg (e.g., 0.1 mg / kg- 10 mg / kg) of antibody is an initial candidate dosage for administration to the patient. A typical daily dosage might range from about 1 pg / kg to 100 mg / kg or more, depending on several factors, e.g., the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. In some embodiments, the antibody or antigen binding fragment thereof described herein or the pharmaceutical composition described herein is administered to the subject at a dose of about 0.1 mg / kg to 1000 mg / kg, about 1 mg / kg to about 500 mg / kg, about 1 mg / kg to about 200 mg / kg, about 1 mg / kg to about 100 mg / kg, or about 0.1 mg / kg to about 50 mg / kg.

[0309] In some embodiments, the antibody or antigen binding fragment thereof disclosed herein or the pharmaceutical composition is administered at a dose of about 100 mg to about 2000 mg, about 200 mg to about 1500 mg, about 300 mg to about 600 mg, about 500 mg to about 1200 mg, or about 300 mg to about 1200 mg. In some embodiments, the antibody, or antigen binding fragment thereof, is administered at a dose of about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg about 1100 mg, about 1200 mg, about 1300 mg, about 1400 mg, about 1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900 mg or about 2000 mg.

[0310] The antibody containing medicament or pharmaceutical composition can be peripherally administered to a subject via a route selected from one or more of: orally, sublingually, buccally, topically, rectally, via inhalation, transdermally, subcutaneously, intravenously, intra-arterially, or intramuscularly, subcutaneously, via intracardiac administration, intraosseously, intradermally,Atorney Docket No. 117823-37620

[0311] intraperitoneally, transmucosally, vaginally, intravitreally, epicutaneously, intra-articularly, peri-articularly, or locally.

[0312] In some embodiments, the antibody, or antigen binding fragment thereof, is administered intramuscularly. In some embodiments, the antibody, or antigen binding fragment thereof, is administered intravenously. In some embodiments, the antibody, or antigen binding fragment thereof, is administered subcutaneously.

[0313] In some embodiments, the antibody, or antigen binding fragment thereof, is administered intramuscularly, intravenously, or subcutaneously at a dose of about 500 mg. In some embodiments, the antibody, or antigen binding fragment thereof, is administered intramuscularly, intravenously, or subcutaneously at a dose of about 600 mg. In some embodiments, the antibody, or antigen binding fragment thereof, is administered intramuscularly, intravenously, or subcutaneously at a dose of about 1200 mg.

[0314] In some embodiments, the antibody or antigen binding fragment thereof described herein or the pharmaceutical composition described herein is administered to the subject intramuscularly, intravenously, or subcutaneously at a dose of about 0.1 mg / kg to 1000 mg / kg, about 1 mg / kg to about 500 mg / kg, about 1 mg / kg to about 200 mg / kg, about 1 mg / kg to about 100 mg / kg, or about 0.1 mg / kg to about 50 mg / kg.

[0315] In one embodiment, the antibody, or antigen binding fragment thereof, is administered once. In one embodiment, the antibody, or antigen binding fragment thereof, is administered weekly. In another embodiment, the antibody, or antigen binding fragment thereof, is administered twice a day, daily, weekly, every two weeks, monthly, every two months, or every three months. In one embodiment, the antibody, or antigen binding fragment thereof, is administered weekly for about four weeks, once weekly for about a month, weekly for about 5 weeks, weekly for about 6 weeks, weekly for about 7 weeks, or weekly for about two months.

[0316] In another aspect, this application provides a method for detecting the presence of GP1 of an arenavirus in a sample in vitro (e.g., a biological sample, such as serum, plasma, tissue, biopsy). The subject method can be used to diagnose a disorder, e.g., arenavirus infection. The method includes: (i) contacting the sample or a control sample with the anti-GPl antibody or fragment thereof as described herein; and (ii) detecting formation of a complex between the anti-GPl antibody or fragment thereof, and the sample or the control sample, wherein a statistically significant change in the formation of the complex in the sample relative to the control sample is indicative of the presence of GP 1 in the sample.

[0317] Given their ability to bind to an GP 1 of an arenavirus, the anti-GP 1 antibodies, or portions thereof, can be used to detect GP1 (e.g., in a biological sample, such as serum or plasma), using a conventional immunoassay, such as an enzyme linked immunosorbent assays (ELISA), an radioimmunoassay (RIA) or tissue immunohistochemistry. In one aspect, the disclosure provides a method for detecting an GP1 of an arenavirus in a biological sample comprising contacting aAtorney Docket No. 117823-37620

[0318] biological sample with an antibody, or antibody portion, and detecting either the antibody (or antibody portion) bound to the GP 1 of an arenavirus or unbound antibody (or antibody portion), to thereby detect GP1 in the biological sample. The antibody is directly or indirectly labeled with a detectable substance to facilitate detection of the bound or unbound antibody. Suitable detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, p-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; and examples of suitable radioactive material include3H14C,35S,90Y, "Tc,inIn,1251,1311,177Lu,166Ho, or153Sm.

[0319] Alternative to labeling the antibody, GP 1 or arenavirus can be assayed in biological fluids by a competition immunoassay utilizing GP1 standards labeled with a detectable substance and an unlabeled anti-GPl antibody. In this assay, the biological sample, the labeled GP1 standards and the anti-GPl antibody are combined and the amount of labeled GP1 standard bound to the unlabeled antibody is determined. The amount of GP1 in the biological sample is inversely proportional to the amount of labeled GP1 standard bound to the anti-GPl antibody. Similarly, GP1 can also be assayed in biological fluids by a competition immunoassay utilizing GP1 standards labeled with a detectable substance and an unlabeled anti-GPl antibody.

[0320] In yet another aspect, this application provides a method for detecting the presence of GP1 or arenavirus in vivo (e.g, in vivo imaging in a subject). The subject method can be used to diagnose a disorder, e.g., an arenavirus-associated disorder. The method includes: (i) administering the anti-GPl antibody or fragment thereof as described herein to a subject or a control subject under conditions that allow binding of the antibody or fragment to GP1; and (ii) detecting formation of a complex between the antibody or fragment and GP1, wherein a statistically significant change in the formation of the complex in the subject relative to the control subject is indicative of the presence of GP1 and / or arenavirus.

[0321] In some embodiments, the antibodies disclosed herein are used or provided in an assay kit and / or method for the detection of arenavirus in mammalian tissues or cells in order to screen / diagnose for a disease or disorder associated with changes in levels of arenavirus. The kit comprises an antibody that binds a GP1 of an arenavirus and means for indicating the binding of the antibody with GP1, if present, and optionally GP1 protein levels. Various means for indicating the presence of an antibody can be used. For example, fluorophores, other molecular probes, or enzymes can be linked to the antibody and the presence of the antibody can be observed in a variety of ways. The method for screening for such disorders can involve the use of the kit, or simply the use of one of the disclosed antibodies and the determination of whether the antibody binds to GP1 in a sample. AsAtorney Docket No. 117823-37620

[0322] will be appreciated by one of skill in the art, high or elevated levels of GP 1 will result in larger amounts of the antibody binding to GP1 in the sample. Thus, degree of antibody binding can be used to determine how much GP1 is in a sample. Subjects or samples with an amount of GP1 that is greater than a predetermined amount (e.g., an amount or range that a person without arenavirus infection or arenavirus-related disorder would have) can be characterized as having an arenavirus-associated disorder.

[0323] The present disclosure further provides for a kit for detecting binding of an anti-GPl antibody of the disclosure to GP1. In particular, the kit may be used to detect the presence of GP1 and / or arenavirus specifically reactive with an anti-GPl antibody of the disclosure or an immunoreactive fragment thereof. The kit may also include an antibody bound to a substrate, a secondary antibody reactive with the antigen and a reagent for detecting a reaction of the secondary antibody with the antigen. Such a kit may be an ELISA kit and can comprise the substrate, primary and secondary antibodies when appropriate, and any other necessary reagents such as detectable moieties, enzyme substrates, and color reagents, for example as described herein. The diagnostic kit may also be in the form of an immunoblot kit. The diagnostic kit may also be in the form of a chemiluminescent kit (Meso Scale Discovery, Gaithersburg, MD). The diagnostic kit may also be a lanthanide-based detection kit (PerkinElmer, San Jose, CA).

[0324] A skilled clinician would understand that a biological sample includes, but is not limited to, sera, plasma, urine, fecal sample, saliva, mucous, pleural fluid, synovial fluid, and spinal fluid.

[0325] V. Pharmaceutical Compositions and Kits

[0326] The present disclosure also provides compositions and kits comprising the antibodies described herein. The compositions and kits may further comprise one or more excipients, carriers, reagents, instructions needed for the use of the antibodies. In some embodiments, the compositions may be pharmaceutical compositions, which comprise the antibodies, derivatives, fragments, analogs and homologs thereof. The pharmaceutical compositions may comprise the antibody and a pharmaceutically acceptable carrier. As used herein, the term “pharmaceutically acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington’s Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference. Suitable examples of such carriers or diluents include water, saline, ringer’s solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.Atorney Docket No. 117823-37620

[0327] A pharmaceutical composition may be formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (e.g., topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application may include one or more of the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH may be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. In some embodiments, any of the antibodies described herein are prepared with carriers that protect against rapid elimination from the body, e.g., sustained and controlled release formulations, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, e.g., ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collage, polyorthoesters, and polylactic acid. Methods for preparation of such pharmaceutical compositions and formulations are apparent to those skilled in the art. For example, the antibodies may be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and polymethylmethacrylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles, and nanocapsules) or in macroemulsions.

[0328] Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g. , films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2- hydroxyethylmethacrylate), or poly(vinylalcohol)), polylactides, copolymers of L-glutamic acid and y ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers (e.g., injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene -vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods.

[0329] In some embodiments, pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration (e.g., in a syringe or intravenous bag), suitable carriers include physiological saline, bacteriostatic water, Cremophor or phosphate buffered saline (PBS). The composition may be sterile and should be fluid toAtorney Docket No. 117823-37620

[0330] the extent that easy syringe ability exists. It may be stable under the conditions of manufacture and storage and preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In some embodiments, isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, and sodium chloride may be included in the composition. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0331] In some embodiments, the pharmaceutical composition may comprise a sterile injectable solution. Sterile injectable solutions may be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by fdtered sterilization. Generally, dispersions may be prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-fdtered solution thereof.

[0332] In some embodiments, the pharmaceutical composition may be formulized for systemic administration. For example, systemic administration may be by intravenous, intradermal, intraperitoneal, intramuscular, or subcutaneously as well by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated may be used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration may be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds may be formulated into ointments, salves, gels, or creams as generally known in the art.

[0333] In some embodiments, the pharmaceutical composition may be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa buter and other glycerides) or retention enemas for rectal delivery.

[0334] In some embodiments, the pharmaceutical composition may be prepared with carriers that protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers may be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolicAtorney Docket No. 117823-37620

[0335] acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.

[0336] It may be advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the disclosure may be dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

[0337] In some embodiments, the compositions (e.g., pharmaceutical compositions) may be included in a container, vial, syringe, injector pen, pack, or dispenser, optionally together with instructions for administration.

[0338] Also provided herein are kits that include any of the antibodies described herein, any of the compositions that include any of the antibodies described herein, or any of the pharmaceutical compositions that include any of the antibodies described herein. Also provided are kits that include one or more second therapeutic agent(s) in addition to an antibody described herein. The second therapeutic agent(s) may be provided in a dosage administration form that is separate from the antibodies. Alternatively, the second therapeutic agent(s) may be formulated together with the antibodies.

[0339] Any of the kits described herein can include instructions for using any of the compositions (e.g., pharmaceutical compositions) and / or any of the antibodies described herein. In some embodiments, the kits can include instructions for performing any of the methods described herein. In some embodiments, the kits can include at least one dose of any of the compositions (e.g., pharmaceutical compositions) described herein. In some embodiments, the kits can provide a syringe for administering any of the pharmaceutical compositions described herein.

[0340] Also provided herein are antibodies produced by any of the methods described herein. Also provided are compositions (e.g., pharmaceutical compositions) that comprise any of the antibodies produced by any of the methods described herein. Also provided herein are kits that include at least one dose of any of the compositions (e.g., pharmaceutical compositions) described herein.

[0341] It will be readily apparent to those skilled in the art that other suitable modifications and adaptations of the methods of the invention described herein may be made using suitable equivalents without departing from the scope of the invention or the embodiments disclosed herein. Having now described the invention in detail, the same will be more clearly understood by reference to the following examples, which are included for purposes of illustration only and are not intended to be limiting.Atorney Docket No. 117823-37620

[0342] EXAMPLES

[0343] Example 1. Broadly Cross-reactive Human Neutralizing Antibodies from Survivors of New World Arenavirus Infection

[0344] To isolate neutralizing antibodies against JUNV from human memory B cells, peripheral blood mononuclear cells (PBMCs) were obtained from four AHF survivors (“AHF1-4”) who were infected with JUNV in Argentina from 1973-2012. A fluorescently labeled version of the JUNV spike protein receptor-binding subunit (GP1) was used as bait in single-B-cell sorting experiments to isolate 29, 20, 118, and 95 GPl-reactive memory B cells from AHF1, AHF2, AHF3, and AHF4, respectively. A total of 26 monoclonal antibodies were recombinantly expressed and purified from AHF1, 14 from AHF2, 92 from AHF3, and 72 from AHF4 (204 antibodies in total). Analysis of antibody sequences from the four AHF survivors revealed enrichment for antibodies that had long CDR H3 loops with a recurrent motif and high frequencies of somatic hypermutation. This signature motif, occurring in 29 / 104 (24%) of neutralizing antibodies, comprised two cysteines separated by four to five amino acids, followed by tyrosine, designated here “CX4-5CY”. Given their enrichment among recovered antibodies, next performed is more extensive analysis of this antibody subset. Nearly all (28 / 29) motif-containing antibodies potently neutralized JUNV pseudotypes, with IC50 values less than 50 ng ml1(Figure 1A).

[0345] To determine whether AHF-derived antibodies have activity against additional pathogenic New World arenaviruses, whether antibodies in the library could cross-neutralize Machupo virus (MACV), Guanarito virus (GTOV), Chapare virus (CHAPV), Sabia virus (SBAV), or SBAV-like virus pseudotypes were tested. Starting with all 104 JUNV-neutralizing antibodies in the library, a cross-neutralization screen of MACV, GTOV, CHAPV, SBAV, or SBAV-like pseudotypes was performed at a concentration of 100 ug ml1. It was found that 17 of the 104 antibodies tested (-16 %) demonstrated activity against JUNV and at least one other New World arenavirus pseudotype (defined as >20% reduction in relative entry at screening). Exactly half of these antibodies were motif-containing and half of the antibodies were non-motif containing (13 total in each group) (Figure IB).

[0346] In the motif-containing group, seven antibodies cross-reacted with at least three New World arenavirus pseudotypes, hereby defined as “broadly cross-reactive” antibodies. The four antibodies that neutralized four pseudotypes were AHF1 B7 (“B7”), AHF2 C5 (“C5”), AHF2 C8 (“C8”), and AHF3 E8.2 (“E8.2”). Three of these antibodies (B7, C5, and C8) all had similar neutralization profiles, with activity of varying degrees against JUNV, MACV, SBAV, and SBAV-like (Figure 2). C8 showed a more unique neutralization profile, neutralizing JUNV, SBAV, SBAV-like, and CHAPV. One motif-containing antibody identified, E8.2, was the only antibody in the library to demonstrate cross-reactivity against five New World arenavirus pseudotypes: JUNV, MACV, SBAV, SBAV-like, and CHAPV (Figure 2). C8 and E8.2 were the only antibodies in the panel of 104Atorney Docket No. 117823-37620

[0347] neutralizing antibodies to demonstrate activity against CHAPV. None of the antibodies we tested had activity against GTOV.

[0348] To provide a structural basis for E8.2 broadly neutralizing activity, the X-ray crystal structure of JUNV GP1 was determined in complex with Fabs E8.2 to a resolution of 2.9 A (Figure 3A). E8.2 buries a surface area of 660.6 2 on the JUNV GP1 with the heavy and light chains each conferring 528 and 132.6 2 respectively. The CDRH3 is responsible for almost all of the heavy chain contact by contributing 96% of the total buried surface area. The paired cysteine residues form a disulfide bridge that supports the extended length of the CDRH3, allowing for flexible but stabilized insertion into the GP1 pocket that is responsible to binding the virus cellular receptor, transferrin receptor 1 (TfRl). The mechanism of neutralization, therefore, is competition for cellular receptor binding.

[0349] To gain insight into how CX4-5CY motif-containing antibodies achieve broad neutralization of multiple New World arenaviruses, we turned to structural studies of the B7 antibody, a prototype antibody of the fourth neutralization profile class. B7 cross-reacts with several New World arenaviruses, specifically, MACV, SBAV, and SBAV-like, despite achieving a neutralization plateau of -50-60% for these viruses (Figure 1A). The next experiment assessed how B7 achieves crossreactivity. To answer this question, the next experiment was performed to determine the X-ray crystal structures of B7 bound to JUNV and MACV GP1 proteins to resolutions of 2.4 and 3 A respectively (Figure 3B-C). B7 buries a total surface area of is 675.2 A2 JUNV GP1 with the VH and VL conferring 641.2 and 34 A2 of surface contact area, respectively. The CDRH3 alone contributes 89% of the total VH BSA. The B7 BSA on MACV interface is 20% larger with the VH and VL accounting for 736.9 and 72.2 A2 respectively. The CDRH3 confers 92% of the entire VH buried surface area.

[0350] Complete amino acid sequences of the heavy and light chains from 4 antibodies are set forth in Table 2, below.

[0351] Table 2. Variable region sequences of antibodies

[0352]

[0353] Atorney Docket No. 117823-37620

[0354]

[0355] Atorney Docket No. 117823-37620

[0356]

[0357] Example 2. In vivo Protective Effect of Antibodies.

[0358] In vivo efficacy of antibodies against Junin Virus Infection. Animal studies are completed under BSL-4 biocontainment. Individual animals are infected with ~ 1,000 pfu of JUNV by i.p. injection. Antibodies or vehicle control are administered i.p. on day 7 and day 11 after infection in volumes of 0.8 to 0.9 mL. 20 mg / kg dose of the antibodies is used in the in vivo study. In another in vivo experiment to determine the breadth of the therapeutic window, groups of guinea pigs are treated with 20 mg / kg dose of antibodies or vehicle control starting on day 6, 7, or 9 after infection and receive a second dose of antibodies 3-4 days later.

[0359] Studies are concluded 28, 30, or 40 d after infection or when animals meet the criteria of sacrifice. Survivals, changes in weights, changes in body temperature, symptoms of neurological diseases (e.g., impaired hind leg use), and plasma viral loads are monitored over the course of the experiment.

[0360] Tissue samples (e.g., liver, spleen, and brain tissue samples) are collected at the endpoint of the experiment for plague assay and immunohistochemistry analysis to assess viral loads and viral antigen levels in tissues, respectively. In addition, the collected tissue samples are processed for histopathology analysis, including hepatocellular vacuolar degeneration, hepatocellular necrosis, Kupffer cell hyperplasia, splenic lymphoid depletion, lymphoplasmacytic meningoencephalitis, and diffuse gliosis. JUNV titration is performed by conventional plaque assay on Vero E6 cells from plasma collected from guinea pigs at various times after infection. Histopathology is performed on liver, spleen, and brain tissue by using Hematoxylin and eosin (H&E) staining and immunohistochemistry. Specific anti-JUNV immunoreactivity is detected by using an anti-JUNV primary antibody. The secondary antibody is biotinylated goat anti-mouse IgG, followed by streptavidin-HRP .Attorney Docket No. 117823-37620

[0361] INCORPORATION BY REFERENCE

[0362] The contents of all references, patents, pending patent applications and published patents, Sequence Listing, figures and Accession Numbers, cited throughout this application are hereby expressly incorporated herein by reference.

[0363] EQUIVALENTS

[0364] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

1. Attorney Docket No. 117823-37620CLAIMS1. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 4 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 8.

2. The antibody, or antigen binding portion thereof, of claim 1, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 3 and a light chain variable region comprising a CDR2 having the amino acid sequence GAS.

3. The antibody, or antigen binding portion thereof, of claim 1 or 2, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 2 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 6.

4. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 12 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 16.

5. The antibody, or antigen binding portion thereof, of claim 4, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 11 and a light chain variable region comprising a CDR2 having the amino acid sequence EVS.

6. The antibody, or antigen binding portion thereof, of claim 4 or 5, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 14.

7. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequenceAttorney Docket No. 117823-37620of SEQ ID NO: 20 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 24.

8. The antibody, or antigen binding portion thereof, of claim 7, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising a CDR2 having the amino acid sequence GAS.

9. The antibody, or antigen binding portion thereof, of claim 7 or 8, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 18 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 22.

10. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 28 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 32.

11. The antibody, or antigen binding portion thereof, of claim 10, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 27 and a light chain variable region comprising a CDR2 having the amino acid sequence DAS.

12. The antibody, or antigen binding portion thereof, of claim 10 or 11, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 26 and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 30.

13. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 3, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 2, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 8, a CDR2 domain comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 6.Attorney Docket No. 117823-3762014. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 11, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 16, a CDR2 domain comprising the amino acid sequence EVS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 14.

15. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 20, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 19, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 18, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 24, a CDR2 domain comprising the amino acid sequence GAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 22.

16. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 28, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 27, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 26, and a light chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 32, a CDR2 domain comprising the amino acid sequence DAS, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 30.

17. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises an amino acid sequence set forth in SEQ ID NO: 1, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 5, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 5.

18. The isolated antibody, or antigen binding portion thereof, of claim 17, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 1 and a light chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 5.Attorney Docket No. 117823-3762019. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 9, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 9, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 13, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13.

20. The isolated antibody, or antigen binding portion thereof, of claim 19, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 13.

21. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 21.

22. The isolated antibody, or antigen binding portion thereof, of claim 21, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 17 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 21.

23. An isolated antibody, or antigen binding portion thereof, that binds to a receptor binding glycoprotein subunit 1 (GP1) of an arenavirus, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, and / or a light chain comprising an amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 29.

24. The isolated antibody, or antigen binding portion thereof, of claim 23, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 29.Attorney Docket No. 117823-3762025. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-24, wherein the GP1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 33-40, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 33-40.

26. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-24, wherein the GP1 is the GP1 of Junin virus (JUNV).

27. The isolated antibody, or antigen binding portion thereof, of claim 26, wherein the GP1 comprises the amino acid sequence set forth in SEQ ID NO: 33 or 34, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 33 or 34.

28. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-24, wherein the GP1 is the GP1 of Machupo virus (MACV).

29. The isolated antibody, or antigen binding portion thereof, of claim 28, wherein the GP1 comprises the amino acid sequence set forth in SEQ ID NO: 35 or 36, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 35 or 36.

30. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-29, wherein the GP1 comprises an epitope comprising an asparagine (D), a serine (S), and a positive charged amino acid residue.

31. The isolated antibody, or antigen binding portion thereof, of claim 30, wherein the positive charged amino acid residue is selected from group consisting of lysine (K) and arginine (R).

32. The isolated antibody, or antigen binding portion thereof, of claim 30 or 31, wherein the GP1 comprises an epitope comprising a serine (S), an asparagine (D), an isoleucine (I), a valine (V), a tyrosine (Y), an lysine (K), and a glutamic acid (E).

33. The isolated antibody, or antigen binding portion thereof, of any one of claims 30-32, wherein the GP1 comprises an epitope comprising amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 19, Y122, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

34. The isolated antibody, or antigen binding portion thereof, of any one of claims 30-33, wherein the GP1 comprises an epitope comprising amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 19, Y122, K169, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.Attorney Docket No. 117823-3762035. The isolated antibody, or antigen binding portion thereof, of any one of claims 30-33, wherein the GP1 comprises an epitope comprising amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 18, LI 19, E121, Y122, D123, R165, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

36. The isolated antibody, or antigen binding portion thereof, of any one of claims 30-32, wherein the GP1 comprises an epitope comprising amino acid residues S97, F98, Rill, V112, S 113, D114, Il 15, SI 16, VI 17, Ml 19, Y122, R165, K169, K170, E171, N174, T216, N218, F226, and Y228 of the amino acid sequence set forth in SEQ ID NO: 35.

37. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-36, wherein the heavy chain variable region comprises a CDR3 that contributes at least about 70% of the contact area between the isolated antibody, or antigen binding portion thereof, and GP 1.

38. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-37, wherein the heavy chain variable region comprises a CDR3 that contributes at least about 75% of the contact area between the isolated antibody, or antigen binding portion thereof, and GP 1.

39. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-38, wherein the antibody, or antigen binding fragment thereof, competes for binding with transferrin receptor 1 (TfRl).

40. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-39, wherein the isolated antibody, or antigen binding fragment thereof, cross-reacts with two or more arenaviruses selected from the group consisting of JUNV, MACV, Sabia virus (SBAV), SBAV-like virus (SBAV-like), and Chapare virus (CHAPV).

41. The isolated antibody, or antigen binding portion thereof, of any one of claims 1-39, wherein the isolated antibody, or antigen binding fragment thereof, cross-reacts with two or more arenaviruses selected from the group consisting of JUNV, SBAV, and SBAV-like.

42. The isolated antibody, or antigen binding portion thereof, of claim 41, wherein the isolated antibody, or antigen binding fragment thereof, further cross-reacts with at least one of MACV and CHAPV.Attorney Docket No. 117823-3762043. The isolated antibody, or antigen binding fragment thereof, of any one of claims 1-42, wherein the antibody, or antigen binding fragment thereof, neutralizes JUNV at an IC50 of about 100 ng / mL or lower, in vitro.

44. The isolated antibody, or antigen binding fragment thereof, of any one of claims 1-43, wherein the antibody, or antigen binding fragment thereof, neutralizes JUNV at an IC50 of about 50 ng / mL or lower, in vitro.

45. The isolated antibody, or antigen binding fragment thereof, of any one of claims 1-44, wherein the antibody, or antigen binding fragment thereof, comprises a Fab, Fab2, or scFv.

46. The isolated antibody, or antigen binding fragment thereof, of any one of claims 1-45, wherein the antibody, or antigen binding fragment thereof, comprises a constant region, an Fc region, or at least one domain thereof.

47. The isolated antibody, or antigen binding fragment thereof, of claim 46, wherein the constant region or Fc region comprises a mutation which impairs at least one effector function.

48. The isolated antibody, or antigen binding fragment thereof, of claim 47, wherein the effector function is selected from the group consisting of FcR binding, complement binding, glycosylation, complement-dependent cytotoxicity (CDC), and antibody-dependent cellular cytotoxicity (ADCC).

49. The isolated antibody, or antigen binding fragment thereof, of claim 46 or 47, wherein the constant or Fc region is a human constant or Fc region.

50. The isolated antibody, or antigen binding fragment thereof, of claim 49, wherein the human constant or Fc region is selected from the group consisting of a human IgGl constant region , a human IgG2 constant region, a human IgG3 constant region, a human IgG4 constant region, and a human Fc constant region.

51. The isolated antibody, or antigen binding fragment thereof, of any one of claims 1-50, wherein the antibody is a human antibody.

52. The isolated antibody, or antigen binding fragment thereof, of any one of claims 1-50, wherein the antibody is a humanized antibody.Attorney Docket No. 117823-3762053. An isolated antibody, or antigen binding fragment thereof, which binds the same epitope as the isolated antibody, or antigen binding fragment thereof, of any one of claims 1-52.

54. The isolated antibody, or antigen binding portion thereof, of claim 53, wherein the heavy chain variable region comprises a CDR3 that comprises a CXXXXCY motif, wherein X is any amino acid residue.

55. The isolated antibody, or antigen binding portion thereof, of claim 53, wherein the heavy chain variable region comprises a CDR3 that comprises a CXXXXXCY motif, wherein X is any amino acid residue.

56. The isolated antibody, or antigen binding portion thereof, of any one of claims 53-55, wherein the epitope comprises an asparagine (D), a serine (S), and a positive charged amino acid residue.

57. The isolated antibody, or antigen binding portion thereof, of claim 56, wherein the positive charged amino acid residue is selected from group consisting of lysine (K) and arginine (R).

58. The isolated antibody, or antigen binding portion thereof, of claim 56 or 57, wherein the GP1 comprises an epitope comprising a serine (S), an asparagine (D), an isoleucine (I), a valine (V), a tyrosine (Y), an lysine (K), and a glutamic acid (E).

59. The isolated antibody, or antigen binding portion thereof, of any one of claims 56-58, wherein the GP1 comprises an epitope comprising amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 19, Y122, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

60. The isolated antibody, or antigen binding portion thereof, of any one of claims 56-58, wherein the GP1 comprises an epitope comprising amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 19, Y122, K169, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

61. The isolated antibody, or antigen binding portion thereof, of any one of claims 56-58, wherein the GP1 comprises an epitope comprising amino acid residues SI 11, DI 13, DI 14, Il 15, Al 16, VI 17, LI 18, LI 19, E121, Y122, D123, R165, E171, K216, and Q218 of the amino acid sequence set forth in SEQ ID NO: 33.

62. The isolated antibody, or antigen binding portion thereof, of any one of claims 56-58, wherein the GP1 comprises an epitope comprising amino acid residues S97, F98, Rill, V112, S 113, D114,Attorney Docket No. 117823-37620Il 15, SI 16, VI 17, Ml 19, Y122, R165, K169, K170, E171, N174, T216, N218, F226, and Y228 of the amino acid sequence set forth in SEQ ID NO: 35.

63. The isolated antibody, or antigen binding portion thereof, of any one of claims 53-62, wherein the heavy chain variable region comprises a CDR3 that contributes at least about 70% of the contact area between the isolated antibody, or antigen binding portion thereof, and the GP1.

64. The isolated antibody, or antigen binding portion thereof, of any one of claims 53-63, wherein the heavy chain variable region comprises a CDR3 that contributes at least about 75% of the contact area between the isolated antibody, or antigen binding portion thereof, and the GP1.

65. The isolated antibody, or antigen binding portion thereof, of any one of claims 53-64, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 4.

66. The isolated antibody, or antigen binding portion thereof, of any one of claims 53-64, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 12.

67. The isolated antibody, or antigen binding portion thereof, of any one of claims 53-64, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 20.

68. The isolated antibody, or antigen binding portion thereof, of any one of claims 53-64, wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 28.

69. An isolated nucleic acid encoding an antibody, or antigen binding portion thereof, of any one of claims 1-68.

70. A pharmaceutical composition comprising the antibody, or antigen binding portion thereof, of any one of claims 1-68, and a pharmaceutically acceptable carrier.

71. A method of treating an infection by one or more arenaviruses, or treating a condition, symptom, disease, or disorder associated with the infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an antibody, or antigenAttorney Docket No. 117823-37620binding fragment thereof, of any of claims 1-68, or a pharmaceutical composition of claim 70, thereby treating the subject.

72. The method of claim 71, wherein the condition, symptom, disease, or disorder is selected from the group consisting of fever, malaise, headache, conjunctivitis, nausea, vomiting, diarrhea, thrombocytopenia, bleeding, neurological disorders, petechiae (such as petechiae in oral mucosa, chest, arms, or axillary region), ataxia, tremors, seizures, and coma.

73. A method of preventing an infection by one or more arenaviruses in a subject in need thereof, the method comprising administering to the subject a prophylactically effective amount of an antibody, or antigen binding fragment thereof, of any of claims 1-68, or a pharmaceutical composition of claim 70, thereby preventing infection by one or more arenaviruses in the subject.

74. A method of inducing an immune response against one or more arenaviruses in a subject in need thereof, the method comprising administering to the subject an effective amount of an antibody, or antigen binding fragment thereof, of any of claims 1-68, or a pharmaceutical composition of claim 70, thereby inducing an immune response against one or more arenaviruses in the subject.

75. The method according to claim 74, wherein the immune response elicits immunoprotection against the one or more arenaviruses.

76. A method of inhibiting or blocking infection of susceptible cells by one or more arenaviruses in a subject in need thereof, the method comprising administering to the subject an effective amount of an antibody, or antigen binding fragment thereof, of any of claims 1-68, or a pharmaceutical composition of claim 70, thereby inhibiting or blocking infection of susceptible cells by one or more arenaviruses in the subject.

77. A method of preventing the onset of a hemorrhagic fever in a subject infected by one or more arenaviruses, or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in a subject infected by one or more arenaviruses, the method comprising administering to the subject a prophylactically or therapeutically effective amount of an antibody, or antigen binding fragment thereof, of any of claims 1-68, or a pharmaceutical composition of claim 70, thereby preventing the onset of a hemorrhagic fever or treating a hemorrhagic fever and / or the symptoms of a hemorrhagic fever in the subject.Attorney Docket No. 117823-3762078. The method of any one of claims 71-77, wherein the one or more arenaviruses are selected from the group consisting of Junin virus (JUNV), Machupo virus (MACV), Sabia virus (SBAV), SBAV-like virus (SBAV-like), and Chapare virus (CHAPV).

79. The method of any one of claims 71-78, wherein the one or more arenaviruses comprise JUNV.

80. The method of any one of claims 71-79, wherein the one or more arenaviruses comprise JUNV, SBAV, and SBAV-like.

81. The method of claim 80, wherein one or more arenaviruses further comprise one or more of MACV and CHAPV.

82. The method of claim 77, wherein the hemorrhagic fever is selected from the group consisting of Argentine hemorrhagic fever, Bolivian hemorrhagic fever, Brazilian hemorrhagic fever, and hemorrhagic fever caused by CHAPV.

83. The method of any one of claims 71-82, wherein the subject is a human subject.

84. The method of any one of claims 71-83, wherein the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject intravenously, intramuscularly, or subcutaneously.

85. The method of any one of claims 71-84, wherein the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject at a dose of about 100 mg to about 2000 mg, about 200 mg to about 1500 mg, about 300 mg to about 600 mg, about 500 mg to about 1200 mg, or about 300 mg to about 1200 mg.

86. The metho of any one of claims 71-84, wherein the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject at a dose of about 0.1 mg / kg to 1000 mg / kg, about 1 mg / kg to about 500 mg / kg, about 1 mg / kg to about 200 mg / kg, about 1 mg / kg to about 100 mg / kg, or about 0.1 mg / kg to about 50 mg / kg.

87. The method of any one of claims 71-86, wherein the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject daily, weekly, monthly, every two months, every three months, or every six months.Attorney Docket No. 117823-3762088. The method of any one claims 71-87, wherein the antibody, or antigen binding fragment thereof, or the pharmaceutical composition is administered to the subject in combination with an antiarenavirus agent or an anti-arenavirus therapy.

89. A method of producing the antibody, or antigen binding portion thereof, of any one of claims 1-68, the method comprising expressing the antibody, or antigen binding portion thereof, in a recombinant cell, and isolating the antibody, or antigen binding portion thereof, from the cell.

90. The method of claim 89, further comprising formulating the antibody, or antigen binding portion thereof, isolated from the cell into a pharmaceutical composition.

91. A kit comprising the antibody, or antigen binding portion thereof, of any one of claims 1-68, the nucleic acid of claim 69, or the pharmaceutical composition of claim 70.

92. A syringe comprising the antibody, or antigen binding portion thereof, of any one of claims 1-68, or the pharmaceutical composition of claim 70.

93. An intravenous bag comprising the antibody, or antigen binding portion thereof, of any one of claims 1-68, or the pharmaceutical composition of claim 70.

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