ISOLATED RECOMBINANT ANTIBODY, USE OF THE ANTIBODY AND THE PHARMACEUTICAL COMPOSITION, AND PHARMACEUTICAL COMPOSITION

Description

106 ISOLATED RECOMBINANT ANTIBODY, USE OF THE ANTIBODY AND THE PHARMACEUTICAL COMPOSITION, AND PHARMACEUTICAL COMPOSITION Separated from BR112017015845-0, filed on 01 / 25 / 2016 FIELD OF THE INVENTION

[001] The present invention relates to antibodies that bind to the glycoprotein of the Ebola virus, pharmaceutical compositions comprising these antibodies and methods of using them. FUNDAMENTALS

[002] Ebola virus (EBOV) and related filoviruses cause severe viral hemorrhagic fever in humans and non-human primates, with a mortality rate of up to approximately 90% in human outbreaks. (Murin, CD et al., (2014), Proc Natl Acad Sci USA, 111(48):17182-17187). The immunological mechanisms that mediate protection are under investigation, but to date, no treatment has been approved for human use.

[003] The Ebola virus glycoprotein (GP) is the only protein present on the surface of the virus and in infected cells. It is presumed to be responsible for the binding and fusion of the virus with host cells. GP exists in several forms. These GPs are encoded in two open reading frames. Unedited GP mRNA produces a secreted non-structural soluble GP (sGP) that is synthesized early in the course of infection (Volchkova et al. (1995), Virology 214:421-430; Volchkova, VA et al., (1998), Virology 250:408-414; Sanchez, et al. (1996), Proc Natl Acad Sci USA, 93:3602-3607; Sanchez, et al. (1999) J. Infect. Dis. 179 (suppl. 1, S164)). sGP forms dimers (Volchkova, et al. (1995), Virology 214:421430; Falzarano, D. et al., Chembiochem (2006), 7:1605-1611) and high quantities are detected in the blood of patients and experimentally infected animals (Sanchez, et al. (1996), Proc Natl Acad Sci USA, 93:36023607; Dolnik, O. et al., (2004), EMBO J 23:2175-2184).

[004] Subsequently, during infection, an edited mRNA is generated, the Petition 870260058248, dated 06 / 15 / 2026, page 14 / 143 / 106, which acquired the ability to encode a second open reading frame. This edited mRNA encodes a form of GP that contains a transmembrane (TM) domain that allows this form of GP to be tethered to the cell plasma membrane and incorporated into virions where it serves as the functional host cell receptor binding protein / fusion protein. During the biosynthesis of this form of GP, the protein is proteolytically processed into two products that are held together by disulfide bonds. The amino-terminal product is referred to as GP1 (140 kDa) and the carboxy-terminal cleavage product is referred to as GP2 (26 kDa) (Sanchez, et al. (1998), J. Virol. 72: 6442-6447).

[005] Ebola virus GP (EBOV GP) may be a target for protective antibodies, but the role of antibodies in disease resistance has been controversial. Insignificant serum titers of neutralizing antibodies in convalescent patients, along with inconsistent results in achieving protection with experimental transfer of immune sera to animals, has resulted in speculation about the function of neutralizing antibodies in recovery from infection (Peters, CJ and LeDuc, JW, (1999), J. Infect. Dis. 179 Suppl 1; Mikhailov, VV, (1994), Vopr. Virusol. 39:82; Xu, L. et al., (1998), Nature Med. 4: 37). However, in the most recent Ebola virus outbreak, some patients who contracted the disease and were treated with a cocktail of monoclonal antibodies (ZMapp) specific for viral GP recovered from the disease.Furthermore, other patients who were treated with serum from these patients and from other patients who survived after acquiring the infection also obtained positive results.

[006] Several antibodies that bind to the GP of the Ebola virus have been described (see, for example, U.S. Patents Nos. 6630144, 6875433, 7335356 and 8513391. See also EP1539238, EP2350270 and EP8513391).

[007] Although technological advances have improved the ability to Petition 870260058248, dated 06 / 15 / 2026, p. 15 / 143 / 106, despite the need to produce Ebola virus antigen vaccine compositions, there is still a need to provide additional sources of protection to address emerging Ebola virus strains. Several candidate therapies against the Ebola virus are currently being evaluated, including post-exposure vaccines (Feldman, H, et al. (2007), PLoS Pathog 3(1):e2), small molecule inhibitors (Cote, M. et al. (2011), Nature, 477(7364):344-348; Johansen, LM, et al. (2013), Sci Transl Med 5(190):190ra179; Warren, TK, et al., (2014), Nature, 508(7496):402-405), siRNA-based therapy (Geisbert, TW, et al., (2006), J. Infect Dis. 193(12):1650-1657; Geisbert, TW et al., (2010), Lancet 375(9729):1896-1905), and monoclonal antibodies (Saphire, EO, (2013), Immunotherapy 5(11):1221-1233; Wong, G. et al. (2014), Trends Microbiol. 22(8):456-463; Qiu, X et al., (2014), Hum. Vaccin. Immunother. 10(4):964967).Passive administration of antibodies to non-human primates has proven effective (Dye, JM., et al., (2012), Proc Natl Acad Sci USA 109(13):5034-5039). More recently, a cocktail of three antibodies (ZMapp) is currently being produced in tobacco plants and is under development for human use (Qiu, X. et al., (2014), Nature 514(7520):47-53).

[008] Although the idea of ​​a vaccine composition comprising the antigen of interest (e.g., GP) to generate neutralizing antibodies in a patient is generally considered a good approach, it may not be advantageous to use it in patients who have already been exposed to the virus, as it would take several weeks for the body to react to the vaccine composition. By that point in time, the patient may have already succumbed to the viral infection, depending on the level of care and palliative therapy available. In these patients, or in any patient who is unable to mount an effective antibody response, it may be more beneficial to provide a composition already containing protective antibodies that can target epitopes common to a particular strain of EBOV or to a variety of strains. Petition 870260058248, dated 06 / 15 / 2026, page 16 / 143 / 106

[009] In this sense, there is still a need in the technique to identify new antibodies, which can be used to prevent or treat an infection caused by the Ebola virus. BRIEF SUMMARY OF THE INVENTION

[0010] The present invention provides antibodies and antigen-binding fragments thereof that bind to the glycoprotein (GP) of the Ebola virus (EBOV). The antibodies of the present invention are useful for inhibiting or neutralizing the activity of the Ebola virus. In some embodiments, the antibodies are useful for blocking the binding of the Ebola virus to the host cell and / or for preventing the entry of the Ebola virus into host cells. In some embodiments, the antibodies function by inhibiting cell-to-cell transmission of the virus or by killing Ebola virus-infected cells, reducing the production of pathogenic viruses. In certain embodiments, the antibodies are useful in preventing, treating, or improving at least one symptom of Ebola virus infection in an individual. In certain embodiments, the antibodies can be administered prophylactically or therapeutically to a subject with or at risk of contracting an Ebola virus infection.In certain embodiments, compositions containing at least one antibody of the invention can be administered to an individual for whom a vaccine is contraindicated or for whom a vaccine is less effective, for example, an elderly patient, a very young patient, a patient who may be allergic to any one or more of the components of a vaccine, or an immunocompromised patient who may not be responsive to the immunogens in a vaccine. In certain embodiments, compositions containing at least one antibody of the invention can be administered to medical staff, hospitalized patients or nursing home residents, or other high-risk patients during an Ebola virus outbreak. In certain embodiments, compositions containing at least one antibody of the invention can be administered as a first-line treatment for... Petition 870260058248, dated 06 / 15 / 2026, page 17 / 143 / 106 patients who have already been exposed to the Ebola virus.

[0011] The antibodies of the invention may be complete (e.g., an IgG1 or IgG4 antibody) or may comprise only an antigen-binding portion (e.g., a Fab, F(ab')2 or scFv fragment) and may be modified to affect functionality, for example, to increase persistence in the host to eliminate residual effector functions (Reddy et al. J. Immunol. 164:1925-1933). In certain embodiments, the antibodies may be bispecific.

[0012] In a first aspect, the present invention provides isolated recombinant monoclonal antibodies or antigen-binding fragments thereof that bind specifically to the GP of EBOV.

[0013] In one embodiment, the present invention provides an isolated recombinant antibody or antigen-binding fragment thereof that binds specifically to Ebola virus (EBOV) and / or an Ebola virus glycoprotein (EBOV-GP), wherein the antibody has one or more of the following characteristics: (a) comprises three heavy chain complementarity determining regions (HCDRs) (HCDR1, HCDR2 and HCDR3) contained in any of the selected heavy chain variable region (HCVR) sequences from the group consisting of SEQ ID NOs: 18, 66, 146, 2, 34, 50, 82, 98, 114, 130, 162, 178, 194, 210, 226, 242, 258, 274, 290 and 306; and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained in any of the selected light chain variable region (LCVR) sequences from the group consisting of SEQ ID NOs: 26, 74, 154, 10, 42, 58, 90, 106, 122, 138, 170, 186, 202, 218, 234, 250, 266, 282 and 298; (b) is a fully human monoclonal antibody; (c) binds to EBOV or a virus-like particle (VIP) expressing an EBOV-GP with a dissociation constant (KD) less than 10-7M, as measured in a plasmon resonance assay of Petition 870260058248, dated 06 / 15 / 2026, page 18 / 143 / 106 surface; (d) demonstrates an increase of at least 3 times in the dissociative half-life (1½) at pH 5 or at pH 6 compared to pH 7.4; (e) demonstrates neutralization of the Zaire ebola virus, with IC50 ranging from about 10-11M to about 10-9M; (f) demonstrates binding to cells expressing EBOV-GP that triggers antibody-dependent cellular cytotoxicity; (g) cross-reacts with one or more EBOV strains selected from the group consisting of Zaire.2014, Zaire.1995, Sudan, Bundibugyo and Cote d'Ivoire; (h) binds to soluble GP (sGP); (i) cross-competition with a reference antibody, wherein the reference antibody comprises a heavy chain variable region (HCVR) and an amino acid sequence of the light chain variable region (LCVR) selected from the group consisting of any of the HCVR and LCVR amino acid sequences in Table 1.

[0014] In one embodiment, the present invention provides an isolated recombinant antibody or antigen-binding fragment thereof that specifically binds to EBOV and / or an Ebola virus glycoprotein (EBOV-GP), wherein the antibody has one or more of the following characteristics: (a) is a complete human monoclonal antibody; (b) binds to EBOV or a virus-like particle (VIP) expressing an EBOV GP with a dissociation constant (Kd) less than 10-7M, as measured in a surface plasmon resonance assay; (c) demonstrates an increase of at least 3 times in the dissociative half-life (0½) at pH 5 or at pH 6 compared to pH 7.4; (d) demonstrates the neutralization of the Ebola Zaire virus, with a Petition 870260058248, dated 06 / 15 / 2026, p. 19 / 143 / 106 IC50 ranging from about 10-11M to about 10-9M; (e) demonstrates binding to cells expressing EBOV GP that triggers antibody-dependent cellular cytotoxicity; (f) cross-reacts with one or more EBOV strains selected from the group consisting of Zaire.2014, Zaire.1995, Sudan, Bundibugyo and Cote d'Ivoire; (g) binds to soluble GP (sGP); (h) cross-competes with a reference antibody, wherein the reference antibody comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR) amino acid sequence selected from the group consisting of either of the HCVR and LCVR amino acid sequences in Table 1.

[0015] Exemplary anti-GP antibodies to the present invention are listed in Tables 1 and 2 of the present invention. Table 1 defines the amino acid sequence identifiers of the heavy chain variable regions (HCVRs), light chain variable regions (LCVRs), heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3), and light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) of exemplary anti-GP antibodies to the Ebola virus. Table 2 establishes the nucleic acid sequence identifiers of the HCVRs, LCVRs, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of exemplary anti-GP antibodies to the Ebola virus.

[0016] The present invention provides antibodies or antigen-binding fragments thereof, comprising an HCVR comprising an amino acid sequence selected from any of the HCVR amino acid sequences listed in Table 1 or a sequence substantially similar thereto, with at least 90%, at least 95%, at least 98% or at least 99% sequence identity thereto. Petition 870260058248, dated 06 / 15 / 2026, p. 20 / 143 / 106

[0017] The present invention also provides antibodies or antigen-binding fragments thereof, comprising an LCVR comprising an amino acid sequence selected from any of the LCVR amino acid sequences listed in Table 1 or a sequence substantially similar thereto with at least 90%, at least 95%, at least 98% or at least 99% sequence identity thereto.

[0018] The present invention also provides antibodies or antigen-binding fragments thereof, comprising an HCVR / LCVR amino acid sequence pair (HCVR / LCVR) comprising any of the HCVR amino acid sequences listed in Table 1, paired with any of the LCVR amino acid sequences listed in Table 1. According to certain embodiments, the present invention provides antibodies or antigen-binding fragments thereof, comprising an HCVR / LCVR amino acid sequence pair contained in any of the anti-GP antibodies to Ebola virus specimens listed in Table 1.

[0019] In one embodiment, the isolated antibody or antigen-binding fragment thereof comprises a pair of HCVR / LCVR amino acid sequences selected from the group consisting of SEQ ID Nos: 18 / 26, 66 / 74, 146 / 154, 2 / 10, 34 / 42, 50 / 58, 82 / 90, 98 / 106, 114 / 122, 130 / 138, 162 / 170, 178 / 186, 194 / 202, 210 / 218, 226 / 234, 242 / 250, 258 / 266, 274 / 282, 290 / 298 and 306 / 282.

[0020] In one embodiment, the isolated antibody or antigen-binding fragment comprises: (a) an HCDR1 domain having an amino acid sequence selected from the group consisting of the following SEQ ID NOs: 20, 68, 148, 4, 36, 52, 84, 100, 116, 132, 164, 180, 196, 212, 228, 244, 260, 276, 292 and 308; (b) an HCDR2 domain having an amino acid sequence selected from the group consisting of the following SEQ IDs: 22, 70, 150, 6, 38, 54, 86, Petition 870260058248, dated 06 / 15 / 2026, page 21 / 143 / 106 102, 118, 134, 166, 182, 198, 214, 230, 246, 262, 278, 294 and 310; (c) an HCDR3 domain having an amino acid sequence selected from the group consisting of the following SEQ ID NOs: 24, 72, 152, 8, 40, 56, 88, 104, 120, 136, 168, 184, 200, 216, 232, 248, 264, 280, 296 and 312; (d) an LCDR1 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 28, 76, 156, 12, 44, 60, 92, 108, 124, 140, 172, 188, 204, 220, 236, 252, 268, 284 and 300; (e) an LCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 30, 78, 158, 14, 46, 62, 94, 110, 126, 142, 174, 190, 206, 222, 238, 254, 270, 286 and 302; (f) an LCDR3 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 32, 80, 160, 16, 48, 64, 96, 112, 128, 144, 176, 192, 208, 224, 240, 256, 272, 288 and 304.

[0021] In certain embodiments, the HCVR / LCVR amino acid sequence pair is selected from the group consisting of SEQ ID NOs: 18 / 26 (H1H17139P), 66 / 74 (H1H17161P) and 146 / 154 (H1H17203P).

[0022] The present invention also provides antibodies or antigen-binding fragments thereof, comprising a heavy chain CDR1 (HCDR1) comprising an amino acid sequence selected from any of the HCDR1 amino acid sequences listed in Table 1 or a sequence substantially similar thereto, with at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0023] The present invention also provides antibodies or antigen-binding fragments thereof, comprising a heavy chain CDR2 (HCDR2) comprising an amino acid sequence selected from any of the HCDR2 amino acid sequences listed in Table 1 or a sequence substantially similar thereto, with at least 90%, at least 95%, at least 98% or at least 99% of Petition 870260058248, dated 06 / 15 / 2026, p. 22 / 143 / 106 sequence identity.

[0024] The present invention also provides antibodies or antigen-binding fragments thereof, comprising a heavy chain CDR3 (HCDR3) comprising an amino acid sequence selected from any of the HCDR3 amino acid sequences listed in Table 1 or a sequence substantially similar thereto, with at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0025] The present invention also provides antibodies or antigen-binding fragments thereof, comprising a light chain CDR1 (LCDR1) comprising an amino acid sequence selected from any of the LCDR1 amino acid sequences listed in Table 1 or a sequence substantially similar thereto, with at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0026] The present invention also provides antibodies or antigen-binding fragments thereof, comprising a light chain CDR2 (LCDR2) comprising an amino acid sequence selected from any of the LCDR2 amino acid sequences listed in Table 1 or a sequence substantially similar thereto, with at least 90%, at least 95%, at least 98% or at least 99% sequence identity.

[0027] The present invention also provides antibodies or antigen-binding fragments thereof, comprising a light chain CDR3 (LCDR3) comprising an amino acid sequence selected from any of the LCDR3 amino acid sequences listed in Table 1 or a sequence substantially similar thereto, with at least 90%, at least 95%, at least 98% or at least 99% sequence identity. Petition 870260058248, dated 06 / 15 / 2026, p. 23 / 143 / 106

[0028] The present invention also provides antibodies or antigen-binding fragments thereof comprising an HCDR3 and LCRD3 (HCVR3 / LCVR3) amino acid sequence pair comprising any of the HCDR3 amino acid sequences listed in Table 1, paired with any of the LCDR3 amino acid sequences listed in Table 1. According to certain embodiments, the present invention provides antibodies or antigen-binding fragments thereof comprising an HCDR3 / LCDR3 amino acid sequence pair contained in any of the exemplary anti-GP Ebola virus antibodies listed in Table 1. In certain embodiments, the HCDR3 / LCDR3 amino acid sequence pair is selected from the group consisting of SEQ ID NOs: 24 / 32 (e.g., H1H17139P), 72 / 80 (e.g., H1H17161P), and 152 / 160 (e.g., H1H17203P).

[0029] The present invention also provides antibodies or antigen-binding fragments thereof, comprising a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3) contained in any of the Ebola virus anti-GP antibodies listed in Table 1. In certain embodiments, the amino acid sequence HCDR1-HCDR2-HCDR3-LCDR1-LCDR-LCDR3 is selected from the group consisting of the SEQ ID NOs: 20-22-24-28-30-32 (e.g., H1H17139P), 68-70-72-76-78-80 (e.g., H1H17161P); and 148-150-152-156-158-160 (e.g., H1H17203P).

[0030] In a related embodiment, the present invention provides antibodies or antigen-binding fragments thereof, comprising a set of six CDRs (i.e., HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3) contained in an HCVR / LCVR amino acid sequence pair, as defined in any of the Ebola virus anti-GP antibodies listed in Table 1. For example, the present invention includes antibodies or antigen-binding fragments of Petition 870260058248, dated 06 / 15 / 2026, p. 24 / 143 / 106, comprising the amino acid sequence set HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 contained in a pair of HCVR / LCVR amino acid sequences selected from the group consisting of SEQ ID NOs: 18 / 26 (e.g., H1H17139P), 66 / 74 (e.g., H1H17161P); and 146 / 154 (e.g., H1H17203P). Methods and techniques for identifying CDRs in HCVR and LCVR amino acid sequences are well known in the art and can be used to identify CDRs in the HCVR and / or LCVR amino acid sequences specified and described in this document. Exemplary conventions that can be used to identify the boundaries of CDRs include, for example, Kabat's definition, Chothia's definition, and AbM's definition.In general terms, the Kabat definition is based on sequence variability, the Chothia definition is based on the location of structural loop regions, and the AbM definition is a consensus between the Kabat and Chothia approaches. See, for example, Kabat, “Sequences of Proteins of Immunological Interest”, National Institutes of Health, Bethesda, Md. (1991); Al-Lazikani et al., J. Mol. Biol. 273:927-948 (1997); and Martin et al., Proc. Natl. Acad. Sci. USA 86: 9268-9272 (1989). Public databases are also available for identifying CDR sequences in an antibody.

[0031] The present invention includes Ebola antiviral antibodies with a modified glycosylation pattern. In some embodiments, modification to remove undesirable glycosylation sites may be useful, or an antibody without a fucose moiety present in the oligosaccharide chain, for example, to enhance antibody-dependent cellular cytotoxicity (ADCC) function (see Shield et al. (2002) JBC 277:26733). In other applications, galactosylation modification may be performed in order to modify complement-dependent cytotoxicity (CDC).

[0032] The present invention also provides antibodies and antigen-binding fragments that compete for specific binding to the Ebola virus with Petition 870260058248, dated 06 / 15 / 2026, p. 25 / 143 / 106 an antibody or an antigen-binding fragment thereof comprising the CDRs of an HCVR and the CDRs of an LCVR, wherein each of the HCVR and LCVR has an amino acid sequence selected from the HCVR and LCVR sequences listed in Table 1.

[0033] The present invention also provides antibodies and antigen-binding fragments that cross-compete for binding to the Ebola virus with a reference antibody or with an antigen-binding fragment thereof comprising the CDRs of an HCVR and the CDRs of an LCVR, wherein each of the HCVR and LCVR has an amino acid sequence selected from the HCVR and LCVR sequences listed in Table 1.

[0034] The present invention also provides isolated antibodies and antigen-binding fragments thereof that block the binding of the Ebola virus to, and / or entry into, a host cell.

[0035] In certain embodiments, the antibodies or antigen-binding fragments of the present invention are bispecific, comprising a first binding specificity to a first epitope on the Ebola virus and a second binding specificity to a second epitope on the Ebola virus, wherein the first and second epitopes are distinct and non-overlapping. In certain embodiments, the bispecific may comprise a first arm that binds to an epitope on the viral glycoprotein and a second arm that binds to an epitope on a different viral antigen.

[0036] In a second aspect, the present invention provides nucleic acid molecules encoding Ebola antiviral antibodies or portions thereof. For example, the present invention provides nucleic acid molecules encoding any of the HCVR amino acid sequences listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCVR nucleic acid sequences listed in Table 2 or a Petition 870260058248, dated 06 / 15 / 2026, p. 26 / 143 / 106 substantially similar sequence to the same having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with them.

[0037] The present invention also provides nucleic acid molecules encoding any of the LCVR amino acid sequences listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCVR nucleic acid sequences listed in Table 2 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with them.

[0038] The present invention also provides nucleic acid molecules encoding any of the HCDR1 amino acid sequences listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCDR1 nucleic acid sequences listed in Table 2 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with them.

[0039] The present invention also provides nucleic acid molecules encoding any of the HCDR2 amino acid sequences listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCDR2 nucleic acid sequences listed in Table 2 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with them.

[0040] The present invention also provides nucleic acid molecules encoding any of the HCDR amino acid sequences. Petition 870260058248, dated 06 / 15 / 2026, p. 27 / 143 / 106 listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCDR3 nucleic acid sequences listed in Table 2 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with them.

[0041] The present invention also provides nucleic acid molecules encoding any of the LCDR1 amino acid sequences listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCDR1 nucleic acid sequences listed in Table 2 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with them.

[0042] The present invention also provides nucleic acid molecules encoding any of the LCDR2 amino acid sequences listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCDR2 nucleic acid sequences listed in Table 2 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with them.

[0043] The present invention also provides nucleic acid molecules encoding any of the LCDR3 amino acid sequences listed in Table 1; in certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the LCDR3 nucleic acid sequences listed in Table 2 or a substantially similar sequence thereof having at least 90%, at least 95%, at least 98% or at least 99% sequence identity with Petition 870260058248, dated 06 / 15 / 2026, p. 28 / 143 / 106 them.

[0044] The present invention also provides nucleic acid molecules encoding an HCVR, wherein the HCVR comprises a set of three CDRs (i.e., HCDR1-HCDR2-HCDR3), wherein the amino acid sequence HCDR1-HCDR2-HCDR3 is as defined by any of the anti-GP antibodies to Ebola virus listed in Table 1.

[0045] The present invention also provides nucleic acid molecules encoding an LCVR, wherein the LCVR comprises a set of three CDRs (i.e., LCDR1-LCDR2-LCDR3), wherein the amino acid sequence LCDR1-LCDR2-LCDR3 is as defined by any of the anti-GP antibodies to Ebola virus listed in Table 1.

[0046] The present invention also provides nucleic acid molecules encoding both an HCVR and an LCVR, wherein the HCVR comprises an amino acid sequence from any of the HCVR amino acid sequences listed in Table 1, and wherein the LCVR comprises an amino acid sequence from any of the LCVR amino acid sequences listed in Table 1. In certain embodiments, the nucleic acid molecule comprises a polynucleotide sequence selected from any of the HCVR nucleic acid sequences listed in Table 2 or a sequence substantially similar thereto with at least 90%, at least 95%, at least 98% or at least 99% sequence identity thereto, and a polynucleotide sequence selected from any of the LCVR nucleic acid sequences listed in Table 2 or a sequence substantially similar thereto with at least 90%, at least 95%, at least 98% or at least 99% sequence identity thereto.In certain embodiments according to this aspect of the invention, the nucleic acid molecule encodes an HCVR and an LCVR, wherein the HCVR and LCVR are both derived from the same anti-GP antibody of the Ebola virus listed in Table 1. Petition 870260058248, dated 06 / 15 / 2026, p. 29 / 143 / 106

[0047] The present invention provides nucleic acid molecules encoding any of the heavy chain amino acid sequences listed in Table 1. The present invention also provides nucleic acid molecules encoding any of the light chain amino acid sequences listed in Table 1.

[0048] In a related aspect, the present invention provides recombinant expression vectors capable of expressing a polypeptide comprising a variable region of the heavy or light chain of an anti-GP antibody of the Ebola virus. For example, the present invention includes recombinant expression vectors comprising any of the nucleic acid molecules mentioned above, i.e., nucleic acid molecules encoding any of the HCVR, LCVR and / or CDR sequences, as defined in Table 1. Also included within the scope of the present invention are the host cells into which such vectors have been introduced, as well as methods for producing the antibodies or portions thereof by culturing the host cells under conditions that permit the production of the antibodies or antibody fragments, and the recovery of the antibodies and antibody fragments produced in this way.

[0049] In a third aspect, the invention provides a pharmaceutical composition comprising one or more isolated monoclonal antibodies or antigen-binding fragments thereof, which bind specifically to the GP of the Ebola virus and a pharmaceutically acceptable carrier or diluent. One or more isolated antibodies comprise a pair of HCVR / LCVR amino acid sequences selected from the group consisting of the HCVR and LCVR sequences listed in Table 1. In one embodiment, the HCVR / LCVR amino acid sequence pair is selected from the group consisting of the SEQ ID Nos: 18 / 26, 66 / 74, 146 / 154, 2 / 10, 34 / 42, 50 / 58, 82 / 90, 98 / 106, 114 / 122, 130 / 138, 162 / 170, 178 / 186, 194 / 202, 210 / 218, 226 / 234, 242 / 250, 258 / 266, 274 / 282, Petition 870260058248, dated 06 / 15 / 2026, p. 30 / 143 / 106 290 / 298 and 306 / 282. In one embodiment, the amino acid sequence pair HCVR / LCVR is selected from the group consisting of the SEQ ID NOs: 18 / 26, 66 / 74 and 146 / 154.

[0050] In a related aspect, the invention provides a composition, which is a combination of at least two antibodies of the invention and a pharmaceutically acceptable vehicle or diluent.

[0051] In a related aspect, the invention provides a composition, which is a combination / cocktail of at least three antibodies of the invention and a pharmaceutically acceptable vehicle or diluent.

[0052] In one embodiment, the pharmaceutical composition comprises (a) a first Ebola antiviral antibody comprising an HCVR / LCVR amino acid sequence pair as described in Table 1 or an antigen-binding fragment thereof; (b) a second Ebola antiviral antibody comprising an HCVR / LCVR amino acid sequence pair as described in Table 1 or an antigen-binding fragment thereof; and (c) a third Ebola antiviral antibody comprising an HCVR / LCVR amino acid sequence pair as described in Table 1 or an antigen-binding fragment thereof, wherein the first antibody binds to or interacts with a first epitope in the Ebola virus GP, and the second and / or third antibody binds to or interacts with a different epitope in the Ebola virus GP, and (d) a pharmaceutically acceptable vehicle or diluent.

[0053] In another related aspect, the invention presents a composition, which is a combination of an anti-GP antibody of the Ebola virus and a second therapeutic agent.

[0054] In one embodiment, the second therapeutic agent is any agent that is advantageously combined with an anti-GP antibody to the Ebola virus. Exemplary agents that can be advantageously combined with an Ebola virus antibody include, without limitation, other agents that... Petition 870260058248, dated 06 / 15 / 2026, page 31 / 143 / 106 bind to and / or inhibit the activity of the Ebola virus (including other antibodies or antigen-binding fragments thereof, etc.) and / or agents that do not bind directly to the Ebola virus but nevertheless inhibit viral activity, including the infectivity of host cells.

[0055] In certain embodiments, the invention provides a pharmaceutical composition comprising: (a) a first Ebola antiviral antibody comprising an HCVR / LCVR amino acid sequence pair, as described in Table 1, or an antigen-binding fragment thereof; (b) a second Ebola antiviral antibody comprising an HCVR / LCVR amino acid sequence pair, as described in Table 1, or an antigen-binding fragment thereof, wherein the first antibody binds to a first epitope in the Ebola virus GP and the second antibody binds to a second epitope in the Ebola virus GP, wherein the first and second epitopes are distinct and non-overlapping; and (c) a pharmaceutically acceptable carrier or diluent.

[0056] In certain embodiments, the invention provides a pharmaceutical composition comprising: (a) a first Ebola antiviral antibody or antigen-binding fragment thereof; (b) a second Ebola antiviral antibody or antigen-binding fragment thereof, wherein the first antibody does not cross-compete with the second antibody for binding to the Ebola virus; and (c) a pharmaceutically acceptable carrier or diluent.

[0057] In certain embodiments, the invention provides a pharmaceutical composition comprising: (a) a first Ebola antiviral antibody or antigen-binding fragment thereof; (b) a second Ebola antiviral antibody or antigen-binding fragment thereof, which interacts with a different Ebola virus antigen, wherein the first antibody binds to an epitope on the GP of the Ebola virus and the second antibody binds to an epitope on a different Ebola virus antigen; and (c) a carrier Petition 870260058248, dated 06 / 15 / 2026, page 32 / 143 / 106 or pharmaceutically acceptable diluent.

[0058] In certain embodiments, the invention provides a pharmaceutical composition comprising: (a) a first Ebola antiviral antibody or an antigen-binding fragment thereof; (b) a second Ebola antiviral antibody or an antigen-binding fragment thereof; (c) a third Ebola antiviral antibody or an antigen-binding fragment thereof, wherein the first, second and / or third epitopes are distinct and non-superimposable; and (d) a pharmaceutically acceptable vehicle or diluent.

[0059] In certain embodiments, the invention provides a pharmaceutical composition comprising: (a) a first Ebola antiviral antibody or antigen-binding fragment thereof; (b) a second Ebola antiviral antibody or antigen-binding fragment thereof; (c) a third Ebola antiviral antibody or antigen-binding fragment thereof, wherein the first antibody may or may not cross-compete with the second and / or third antibody for binding to the Ebola virus; and (d) a pharmaceutically acceptable carrier or diluent.

[0060] In certain embodiments, the invention provides a pharmaceutical composition comprising: (a) a first Ebola antiviral antibody or antigen-binding fragment thereof; (b) a second and / or a third Ebola antiviral antibody or antigen-binding fragment thereof, which interacts with different Ebola virus antigens, wherein the first antibody binds to an epitope on the Ebola virus and the second and / or third antibody binds to an epitope on a different Ebola virus antigen; and (c) a pharmaceutically acceptable vehicle or diluent.

[0061] In one embodiment, the pharmaceutical composition comprises a first Ebola antiviral antibody or an antigen-binding fragment thereof that binds to or interacts with an epitope on an Ebola virus strain, and a second and / or third Ebola antiviral antibody or an antigen-binding fragment thereof that binds to or interacts with an Petition 870260058248, dated 06 / 15 / 2026, p. 33 / 143 / 106 second and / or third epitope in the same strain or in a different strain of the Ebola virus. The Ebola virus strains that interact with an antibody of the invention may be selected from the group consisting of the Zaire.2014, Zaire.1995, Sudan, Bundibugyo, and Cote d'Ivoire strains or variants thereof.

[0062] In a related aspect, the invention provides a pharmaceutical composition comprising a first isolated monoclonal antibody or an antigen-binding fragment thereof that specifically binds to the GP of the Ebola virus, wherein the first isolated monoclonal antibody or an antigen-binding fragment thereof comprises an amino acid sequence HCDR1 of SEQ ID NO: 20; an amino acid sequence HCDR2 of SEQ ID NO: 22; an amino acid sequence HCDR3 of SEQ ID NO: 24; an amino acid sequence LCDR1 of SEQ ID NO: 28; an amino acid sequence LCDR2 of SEQ ID NO: 30 and an amino acid sequence LCDR3 of SEQ ID NO: 32, and a pharmaceutically acceptable vehicle or diluent.The pharmaceutical composition may further comprise a second isolated monoclonal antibody or an antigen-binding fragment thereof that specifically binds to the GP of the Ebola virus, wherein the second isolated monoclonal antibody or an antigen-binding fragment thereof comprises an amino acid sequence HCDR1 from SEQ ID NO: 68; an amino acid sequence HCDR2 from SEQ ID NO: 70; an amino acid sequence HCDR3 from SEQ ID NO: 72; an amino acid sequence LCDR1 from SEQ ID NO: 76; an amino acid sequence LCDR2 from SEQ ID NO: 78 and an amino acid sequence LCDR3 from SEQ ID NO: 80. The pharmaceutical composition may further comprise a third isolated monoclonal antibody or an antigen-binding fragment thereof that specifically binds to the GP of the Ebola virus, wherein the third isolated monoclonal antibody or an antigen-binding fragment thereof comprises an amino acid sequence HCDR1 from SEQ ID NO: 148; an amino acid sequence. Petition 870260058248, dated 06 / 15 / 2026, p. 34 / 143 / 106 HCDR2 from SEQ ID NO: 150; an amino acid sequence HCDR3 from SEQ ID NO: 152; an amino acid sequence LCDR1 from SEQ ID NO: 156; an amino acid sequence LCDR2 from SEQ ID NO: 158 and an amino acid sequence LCDR3 from SEQ ID NO: 160.

[0063] In certain embodiments, each antibody may be formulated as a separate formulation and, if it is determined that more than one antibody is needed to achieve maximum therapeutic efficacy, each of the antibody formulations may be co-administered (simultaneously or sequentially) as needed. Alternatively, the antibody cocktail may be co-formulated.

[0064] In certain embodiments, when two or more antibodies are combined in a pharmaceutical composition, they may or may not bind to the same epitope or to overlapping epitopes on the Ebola virus protein. Additional combination therapies and co-formulations involving the Ebola antiviral antibodies of the present invention are described in other sections of the present invention.

[0065] In a fourth aspect, the invention provides therapeutic methods for treating a disease or disorder associated with the Ebola virus (such as viral infection in an individual) or at least one symptom associated with viral infection or the frequency or severity of at least one symptom associated with EBOV infection, using an anti-GP antibody of the Ebola virus or antigen-binding portion of an antibody of the invention or a cocktail of at least two or more antibodies of the invention, wherein the therapeutic methods comprise administering a therapeutically effective amount of at least two or more antibodies or antigen-binding fragments of the invention to the individual in need thereof. In one embodiment, the methods comprise administering a combination (cocktail) of at least three antibodies of the invention. In one embodiment, the antibody cocktail comprises three anti-EBOV antibodies, Petition 870260058248, dated 06 / 15 / 2026, page 35 / 143 / 106, having the amino acid sequence pairs as defined in SEQ ID Nos: 18 / 26, 66 / 74 and 146 / 154. The treated disorder is any disease or condition that is improved, alleviated, inhibited or prevented through the inhibition of Ebola virus activity. In certain embodiments, the invention provides methods for preventing, treating or improving at least one symptom of Ebola virus infection, the method comprising administering a therapeutically effective amount of at least one or more anti-GP antibodies to the Ebola virus or antigen-binding fragments thereof of the invention to an individual in need thereof.

[0066] In a related aspect, the invention provides a method for neutralizing infectious EBOV, the method comprising exposing an EBOV-infected cell to a composition comprising one or more anti-EBOV antibodies or antigen-binding fragments thereof, wherein the exposure results in increased protection of the cell against viral infection or cell death. In certain embodiments, the exposure may be in vitro or in vivo. In one embodiment, the methods comprise administering one or more antibodies of the invention. In one embodiment, the methods comprise administering a combination (cocktail) of at least three antibodies of the invention. In one embodiment, the antibody cocktail comprises three anti-EBOV antibodies having amino acid sequence pairs as defined in SEQ ID Nos: 18 / 26, 66 / 74 and 146 / 154.

[0067] In some embodiments, the present invention provides methods for improving or reducing the severity, duration, or frequency of occurrence of at least one symptom of Ebola virus infection in an individual by administering one or more anti-GP antibodies to the Ebola virus of the invention, wherein at least one symptom is selected from the group consisting of fever, headache, fatigue, loss of appetite, myalgia, diarrhea, vomiting, abdominal pain, dehydration, and unexplained bleeding.

[0068] In certain embodiments, the invention provides methods for Petition 870260058248, dated 06 / 15 / 2026, page 36 / 143 / 106 to reduce the viral load in an individual, the methods comprising administering to the individual an effective amount of one or more antibodies or fragments thereof of the invention, which binds to the GP of the Ebola virus and blocks the binding of the Ebola virus and / or entry into the host cell.

[0069] In a related aspect, the invention provides a method of increasing the survival or probability of survival of an individual suffering from infection caused by EBOV or an individual exposed to EBOV or at risk from exposure to the virus or of acquiring EBOV, the method comprising administering at least one antibody or antigen-binding fragment of the invention or a pharmaceutical composition comprising at least one antibody of the invention to an individual in need thereof.

[0070] In one embodiment, the invention provides a method of increasing the survival or probability of survival of an individual suffering from EBOV infection or an individual exposed to EBOV or at risk of exposure to or contracting EBOV, the method comprising administering an antibody cocktail comprising a mixture of at least two anti-EBOV antibodies of the invention. In one embodiment, the method comprises administering an antibody cocktail comprising a mixture of at least three anti-EBOV antibodies of the invention. In one embodiment, the antibody cocktail to be administered comprises a mixture of at least three anti-EBOV antibodies of the invention, wherein the at least three antibodies comprise amino acid sequence pairs HCVR / LCVR, as defined in SEQ ID Nos: 18 / 26, 66 / 74 and 146 / 154.

[0071] In one embodiment, the subject that requires this is an individual at risk of exposure to or acquisition of an Ebola virus infection, wherein the individual is selected from the group consisting of an immunocompromised individual, a healthcare worker, a person who is Petition 870260058248, dated 06 / 15 / 2026, page 37 / 143 / 106 suspected of having been exposed to a person infected with the Ebola virus, a person who comes into physical contact or close physical proximity with an infected individual, a hospital worker, a pharmaceutical researcher, maintenance staff responsible for cleaning the facilities of a hospital or institution where a patient with Ebola was treated, individuals who have visited or are planning to visit an area or country known to have or suspected of having an outbreak of the Ebola virus, and a frequent airline passenger.

[0072] In one embodiment, the individual in need may be administered at least one anti-EBOV antibody of the invention or an antigen-binding fragment thereof, or a pharmaceutical composition comprising at least one antibody or antigen-binding fragment thereof of the invention, together with a second therapeutic agent. The second therapeutic agent may be selected from the group consisting of an antiviral drug, an anti-inflammatory drug (such as corticosteroids and nonsteroidal anti-inflammatory drugs), an antibody other than EBOV, an EBOV vaccine, TKM Ebola (small interfering RNAs targeting viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (phosphorodiamidatomorpholino antisense oligomers targeting the VP24 gene of the Ebola virus), and interferons.

[0073] In one embodiment, the pharmaceutical composition may be administered subcutaneously, intravenously, intradermally, intramuscularly, intranasally, or orally.

[0074] In a related embodiment, increased protection can be observed in a mammal exposed to or infected with EBOV when the mammal is treated with a pharmaceutical composition comprising an antibody cocktail, comprising at least three antibodies of the invention. Petition 870260058248, dated 06 / 15 / 2026, page 38 / 143 / 106

[0075] In one embodiment, the enhanced protection observed may be measured by a decrease in the severity or frequency of at least one symptom associated with EBOV infection, by a reduction in viral load, or by an increase in the survival of an EBOV-infected mammal. The at least one symptom may be selected from the group consisting of fever, headache, fatigue, loss of appetite, myalgia, diarrhea, vomiting, abdominal pain, dehydration, and unexplained bleeding.

[0076] Increased protection may be observed when the antibody is used alone or when it is used in combination with one or more additional therapeutic agents or anti-EBOV treatment modalities.

[0077] One or more additional therapeutic agents may be selected from the group consisting of an antiviral drug, an anti-inflammatory drug (such as corticosteroids and nonsteroidal anti-inflammatory drugs), a different antibody to the Ebola virus, a vaccine for the Ebola virus, TKM Ebola (small interfering RNAs that target viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (antisense phosphorodiamidate morpholino oligomers that target the VP24 gene of the Ebola virus) and interferons.

[0078] In one embodiment, one or more additional therapeutic agents include one or more anti-EBOV antibodies.

[0079] In one embodiment, one or more anti-EBOV antibodies comprise an amino acid sequence from the heavy chain variable region (HCVR) and the light chain variable region (LCVR) selected from the group consisting of any of the HCVR and LCVR amino acid sequences from Table 1.

[0080] In a related embodiment, one or more anti-EBOV antibodies comprise a pair of amino acid sequences from the heavy chain variable region (HCVR) and the light chain variable region (LCVR) selected from the group consisting of SEQ ID Nos: 2 / 10, 18 / 26, Petition 870260058248, dated 06 / 15 / 2026, page 39 / 143 / 106 34 / 42, 50 / 58, 66 / 74, 82 / 90, 98 / 106, 114 / 122, 130 / 138, 146 / 154, 162 / 170, 178 / 186, 194 / 202, 210 / 218, 226 / 234, 242 / 250, 258 / 266, 274 / 282, 290 / 298 and 306 / 282.

[0081] In another related embodiment, one or more anti-EBOV antibodies comprise a pair of amino acid sequences, heavy chain variable region (HCVR) and light chain variable region (LCVR), selected from the group consisting of any of the SEQ ID Nos: 18 / 26, 66 / 74 and 146 / 154.

[0082] In certain modalities, one or more antibodies or antigen-binding fragments may be administered prophylactically or therapeutically to an individual who has, or is at risk of having, or is predisposed to developing an Ebola virus infection. Individuals at risk include, but are not limited to, an immunocompromised person, for example, a person who is immunocompromised because of an autoimmune disease or those receiving immunosuppressive therapy (e.g., after organ transplantation) or those afflicted with human immunodeficiency syndrome (HIV) or acquired immunodeficiency syndrome (AIDS), certain forms of anemia that deplete or destroy white blood cells, people receiving radiation or chemotherapy, or those afflicted with an inflammatory disorder.Other individuals at risk of acquiring an infection caused by the Ebola virus include healthcare workers or anyone who comes into physical contact or close proximity with an infected individual or who is exposed to bodily fluids or tissues of infected individuals; they also have a higher risk of developing an Ebola virus infection. Additionally, a subject is at risk of contracting an Ebola virus infection due to proximity to an outbreak of the disease, for example, an individual residing in a densely populated city or near individuals with confirmed or suspected Ebola virus infections, or by choice of employment, for example, maintenance staff. Petition 870260058248, dated 06 / 15 / 2026, page 40 / 143 / 106 responsible for cleaning a hospital facility or institution where an Ebola patient was treated, a hospital worker, a pharmaceutical researcher, an individual who visited or plans to visit an area or country known to have or suspected of having an Ebola virus outbreak, or a frequent airline passenger.

[0083] In certain embodiments, the antibody or antigen-binding fragment thereof of the invention is administered together with a second therapeutic agent to the individual who needs it. The second therapeutic agent may be selected from the group consisting of an anti-inflammatory drug (such as corticosteroids and nonsteroidal anti-inflammatory drugs), an anti-infective drug, an antiviral drug, a different antibody for the Ebola virus, a vaccine for the Ebola virus, ZMapp therapy, Ebola TKM (small interfering RNAs that target viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (phosphorodiamidatomorpholino antisense oligomers that target the VP24 gene of the Ebola virus), interferons, a dietary supplement such as antioxidants, and any other drug or therapy known in the art useful for improving at least one symptom of Ebola virus infection or for reducing the viral load in a patient.In certain embodiments, the second therapeutic agent may be an agent that helps to neutralize or reduce any possible side effect(s) associated with an antibody or antigen-binding fragment of the invention, should such side effect(s) occur. The antibody or fragment thereof may be administered subcutaneously, intravenously, intradermally, intraperitoneally, orally, intranasally, intramuscularly, or intracranially. In one embodiment, the antibody may be administered as a single intravenous infusion to achieve the maximum concentration of the antibody in the individual's serum; the antibody or fragment thereof may be administered at a dose of... Petition 870260058248, dated 06 / 15 / 2026, page 41 / 143 / 106 approximately 0.1 mg / kg of body weight up to approximately 100 mg / kg of body weight of the individual. In certain embodiments, an antibody of the present invention may be administered in one or more doses, comprising between 50 mg and 600 mg.

[0084] The present invention also includes an Ebola antiviral antibody or an antigen-binding fragment thereof of the invention for use in treating an individual who has or is suspected of having or has been exposed to EBOV or for use in manufacturing a medicament for the treatment of a disease or disorder that could benefit from blocking the binding of the Ebola virus and / or its activity.

[0085] Other modalities will become apparent from a review of the detailed description below. BRIEF DESCRIPTION OF THE FIGURES

[0086] Figure 1: H1H17161P potentially neutralizes live EBOV.

[0087] Figure 2: Shows the interaction of the three anti-EBOV antibodies with Ebola GP or soluble Ebola GP (sGP). DETAILED DESCRIPTION

[0088] Before the present methods are described, it should be understood that this invention is not limited to the particular methods and experimental conditions described, since such methods and conditions may vary. It is also to be understood that the terminology used in this document is intended only to describe particular embodiments, and is not intended to limit the scope of the present invention, which will be limited only by the appended claims.

[0089] Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as that commonly understood by a person skilled in the art to which this invention pertains. Although any similar methods and materials or Petition 870260058248, dated 06 / 15 / 2026, page 42 / 143 / 106 equivalent to those described in the present invention may be used in the practice or testing of the present invention; preferred methods and materials will now be described. Definitions

[0090] The “Ebola virus” or “EBOV” is a genus of the family Filoviridae, which is known to cause severe and rapidly progressive hemorrhagic fever. There are many different species and strains of Ebola virus based on the nucleotide sequence and the location of the outbreak, for example, Zaire, Tai Forest (formerly known as Côte d'Ivoire or Ivory Coast), Sudan, Reston, and Bundibugyo. The most lethal forms of the virus are the Zaire and Sudan strains. The Reston strain is the only strain known to infect only non-human primates. The term "Ebola virus" also includes variants of the Ebola virus isolated from different Ebola virus isolates.

[0091] The amino acid sequence of the complete Ebola virus glycoprotein referred to in this document as “GP EBOV” or “GP of the Ebola virus” is exemplified by the amino acid sequences found in GenBank as accession numbers AHX24649.1 (see also SEQ ID NO: 314) and AHX24649.2 (see also SEQ ID NO: 315). The term also encompasses GP of the Ebola virus or a fragment thereof coupled, for example, to a histidine marker (e.g., see registry number AHX24649.1 with a deca-histidine marker (SEQ ID NO: 318)), mouse or human Fc, or a signal sequence. The amino acid sequence of “soluble GP” or “sGP” is shown in registry number AHX24650 and as SEQ ID NO: 316 (with the signal sequence) and also as SEQ ID NO: 317 (without the signal sequence, but with a myc-mychexa-histidine marker). The amino acid sequence of “GP1” begins at the amino-terminal end of the complete GP at residue 1 and ends at residue 501 of SEQ ID NO: 315.The amino acid sequence “GP2” spans residues 502 to 676 of the complete GP, shown as SEQ ID NO: 315. Petition 870260058248, dated 06 / 15 / 2026, page 43 / 143 / 106

[0092] The term “Ebola virus infection” or “EBOV infection,” as used in the present invention, refers to severe hemorrhagic fever resulting from exposure to the virus or an infected animal or an infected human patient, or from contact with the body fluids or tissues of a human or animal patient with an Ebola virus infection. “Symptoms associated with an Ebola virus infection” include fever, headache, fatigue, loss of appetite, myalgia, diarrhea, vomiting, abdominal pain, dehydration, and unexplained bleeding.

[0093] The term “antibody,” as used in this document, refers to immunoglobulin molecules composed of four polypeptide chains, two heavy chains (H) and two light chains (L) linked by disulfide bonds (i.e., complete antibody molecules), as well as their multimers (e.g., IgM) or antigen-binding fragments thereof. Each heavy chain comprises a variable region of the heavy chain (“HCVR” or “VH”) and a constant region of the heavy chain (composed of the CH1, CH2, and CH3 domains). Each light chain comprises a variable region of the light chain (“LCVR” or “VL”) and a constant region of the light chain (Cl). The Vh and Vl regions may be further subdivided into regions of hypervariability, termed complementarity-determining regions (CDRs), interspersed with regions that are more conserved, termed structure regions (FRs).Each Vh and Vl is composed of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1 CDR1, FR2, CDR2, FR3, CDR3, and FR4. In certain embodiments of the invention, the antibody FRs (or the antigen-binding fragment thereof) may be identical to human germline sequences or may be naturally or artificially modified. A consensus amino acid sequence may be defined based on a side-by-side analysis of two or more CDRs. Petition 870260058248, dated 06 / 15 / 2026, p. 44 / 143 / 106

[0094] The substitution of one or more CDR residues or the omission of one or more CDRs is also possible. Antibodies have been described in the scientific literature in which one or two CDRs can be dispensed with for binding. Padlan et al. (1995 FASEB J.) analyzed the contact regions between antibodies and their antigens, based on published crystal structures, and concluded that only about one-fifth to one-third of the CDR residues effectively come into contact with the antigen. Padlan also found that many antibodies in which one or two CDRs had no amino acids in contact with an antigen (see also, Vajdos et al. 2002 J Mol Biol 320:415-428).

[0095] CDR residues that do not come into contact with the antigen can be identified based on previous studies (e.g., the H60-H65 residues in CDRH2 are often not needed), from Kabat CDR regions that lie outside Chothia CDRs, by molecular modeling, and / or empirically. If a CDR or residue(s) thereof are omitted, it is usually replaced with an amino acid that occupies the corresponding position in another human antibody sequence or a consensus of such sequences. Positions for substitution in CDRs and amino acids to be substituted can also be selected empirically. Empirical substitutions can be conservative or non-conservative substitutions.

[0096] The fully human Ebola antiviral monoclonal antibodies described in the present invention may comprise one or more amino acid substitutions, insertions and / or deletions in the structure and / or CDR regions of the variable heavy and light chain domains, compared to the corresponding germline sequences. Such mutations can be easily verified by comparing the amino acid sequences described in this document with available germline sequences, for example, from antibody sequence databases. Petition 870260058248, dated 06 / 15 / 2026, p. 45 / 143 / 106 public. The present invention includes antibodies and antigen-binding fragments thereof, which are derived from any of the amino acid sequences described in this document, wherein one or more amino acids in one or more structure and / or CDR regions are modified to the corresponding residue(s) of the germline sequence from which the antibody was derived or to the corresponding residue(s) of another germline sequence or to a germline residue-conserving amino acid substitution (such sequence alterations are referred to in this invention collectively as “germline mutations”).A person normally skilled in the art, starting from the variable region sequences of the heavy chain and light chain described in the present invention, can easily produce numerous antibodies and antigen-binding fragments comprising one or more germline mutations or combinations thereof. In certain embodiments, all structural and / or CDR residues in the VH and / or VL domains are modified back to the residues found in the original germline sequence from which the antibody was derived. In other embodiments, only certain residues are modified back to the original germline sequence, for example, only the mutant residues found in the first 8 amino acids of FR1 or in the last 8 amino acids of FR4 or only the mutant residues found in CDR1, CDR2 or CDR3.In other embodiments, one or more of the structure and / or CDR residue(s) are modified to the corresponding residue(s) of a different germline sequence (i.e., a germline sequence that is different from the germline sequence from which the antibody was originally derived). Furthermore, the antibodies of the present invention may contain any combination of two or more germline mutations in the structure and / or CDR regions, for example, in... Petition 870260058248, dated 06 / 15 / 2026, page 46 / 143 / 106, states that certain individual residues are modified to the corresponding residue of a particular germline sequence, while certain other residues that differ from the original germline sequence are retained or modified to the corresponding residue of a different germline sequence. Once obtained, antibodies and antigen-binding fragments containing one or more germline mutations can be easily tested for one or more desired properties, such as enhanced binding specificity, enhanced binding affinity, improved or increased biological antagonist or agonist properties (as the case may be), reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained in this manner are generally encompassed in the present invention.

[0097] The present invention also includes fully human Ebola antiviral monoclonal antibodies comprising variants of any of the HCVR, LCVR and / or CDR amino acid sequences described in this invention, with one or more conservative substitutions. For example, the present invention includes Ebola antiviral antibodies with HCVR, LCVR and / or CDR amino acid sequences with, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc. conservative amino acid substitutions relative to any of the HCVR, LCVR and / or CDR amino acid sequences described in that document.

[0098] The term “human antibody,” as used in this document, is intended to include antibodies with variable and constant regions derived from human germline immunoglobulin sequences. The human mAbs of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example, in Petition 870260058248, dated 06 / 15 / 2026, page 47 / 143 / 106 CDRs, and particularly CDR3. However, the term “human antibody,” as used herein, is not intended to include mAbs in which germline-derived CDR sequences from another mammalian species (e.g., mouse) have been grafted onto human FR sequences. The term includes antibodies recombinantly produced in a non-human mammal or in cells of a non-human mammal. The term is not intended to include antibodies isolated from or generated in a human individual.

[0099] The term “recombinant,” as used in the present invention, refers to antibodies or antigen-binding fragments thereof of the invention created, expressed, isolated, or obtained by technologies or methods known in the art as recombinant DNA technology, which include, for example, DNA joining and transgenic expression. The term refers to antibodies expressed in a non-human mammal (including transgenic non-human mammals, for example, transgenic mice) or a cell expression system (for example, CHO cells) or isolated from a recombinant combinatorial human antibody library.

[00100] The term “specifically binds” or “specifically binds to” or similar, means that an antibody, or an antigen-binding fragment thereof, forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding can be characterized by an equilibrium dissociation constant of at least about 1 x 10⁻⁷ M or less (e.g., a smaller KD denotes a stronger binding). Methods for determining whether two molecules bind specifically are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. As described in the present invention, antibodies have been identified by surface plasmon resonance, for example, BIACORE™, that specifically bind to the Ebola virus. Furthermore, multispecific antibodies Petition 870260058248, dated 06 / 15 / 2026, page 48 / 143 / 106 that bind to a domain in the Ebola virus and one or more additional antigens or a bispecific antigen that binds to two different regions of the Ebola virus, nevertheless, are considered antibodies that “bind specifically”, as used in the present invention.

[00101] The term “high-affinity” antibody refers to mAbs with a binding affinity to the Ebola virus, expressed as KD, of at least 10-7M; preferably 10-8M; more preferably 10-9M, even more preferably 10-11M, and even more preferably 10-12M, as measured by surface plasmon resonance, for example, BIACORE™ or solution affinity ELISA.

[00102] The term “slow dissociation rate”, “Koff” or “kd” refers to an antibody that dissociates from the Ebola virus or a virus-like particle expressing the GP of the Ebola virus, with a rate constant of 1 x 10-3s-1 or less, preferably 1 x 10-4s-1 or less, as determined by surface plasmon resonance, for example, BIACORE™.

[00103] The terms “antigen-binding portion” of an antibody, “antigen-binding fragment” of an antibody, and the like, as used in the present invention, include any genetically engineered, synthetic polypeptide or glycoprotein that can be enzymatically obtained or that occurs in nature and that specifically binds to an antigen to form a complex. The terms “antigen-binding fragment” of an antibody or “antibody fragment,” as used in the present invention, refer to one or more fragments of an antibody that retain the ability to bind to the Ebola virus.

[00104] In specific embodiments, antibody or antibody fragments of the invention may be conjugated with a moiety, such as a ligand or a therapeutic moiety (“immunoconjugate”), such as an antiviral drug, a second Ebola antiviral antibody or any other moiety. Petition 870260058248, dated 06 / 15 / 2026, p. 49 / 143 / 106 useful therapy in the treatment of an infection caused by the Ebola virus.

[00105] An “isolated antibody”, as used in this document, is intended to refer to an antibody that is substantially free of other antibodies (Abs) with different antigenic specificities (for example, an isolated antibody that binds specifically to the Ebola virus or a fragment thereof is substantially free of Abs that bind specifically to antigens other than those of the Ebola virus).

[00106] A “blocking antibody” or a “neutralizing antibody,” as used in the present invention (or an “antibody that neutralizes Ebola virus activity” or “antagonist antibody”), is intended to refer to an antibody whose binding to the Ebola virus results in the inhibition of at least one biological activity of the Ebola virus. For example, an antibody of the invention may prevent or block the binding of the Ebola virus to, or entry into, a host cell. Furthermore, a “neutralizing antibody” is one that can neutralize, i.e., prevent, inhibit, reduce, impede, or interfere with, the ability of a pathogen to initiate and / or perpetuate an infection in a host. The terms “neutralizing antibodies” and “an antibody that neutralizes” or “antibodies that neutralize” are used interchangeably in the present invention.These antibodies can be used alone or in combination, as prophylactic or therapeutic agents with other antiviral agents using the appropriate formulation, or in conjunction with active vaccination, or as a diagnostic tool.

[00107] “Antibody-dependent cell-mediated cytotoxicity” or “ADCC” is a cell-mediated immune defense mechanism whereby an effector cell of the immune system actively lyses a target cell whose membrane surface antigens have been bound by specific antibodies, such as those described in the present invention. As such, it is a mechanism by which, for example, a virus-specific antibody can act to limit the spread of infection. ADCC Petition 870260058248, dated 06 / 15 / 2026, page 50 / 143 / 106 classic is mediated by natural killer cells (NK cells), macrophages, neutrophils and, in certain cases, eosinophils.

[00108] The term “surface plasmon resonance”, as used in this document, refers to an optical phenomenon that allows the analysis of biomolecular interactions in real time through the detection of changes in protein concentrations within a biosensor array, for example, using the BIACORE™ system (Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, NJ).

[00109] The term “Kd”, as used in this document, is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[00110] The term “epitope” refers to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule known as a paratope. A single antigen can have more than one epitope. Thus, different antibodies can bind to different areas on an antigen and can have different biological effects. The term “epitope” also refers to a site on an antigen to which B and / or T cells respond. It also refers to a region of an antigen that is bound by an antibody. Epitopes can be defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and have those residues that directly contribute to the affinity of the interaction. Epitopes can also be conformational, i.e., composed of non-linear amino acids.In certain embodiments, epitopes may include determinants that are chemically active surface clusters of molecules, such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and, in certain embodiments, they may have specific three-dimensional structural features and / or specific charge characteristics.

[00111] The term “cross-competes”, as used in Petition 870260058248, dated 06 / 15 / 2026, p. 51 / 143 / 106, the present invention, means an antibody or an antigen-binding fragment thereof that binds to an antigen and inhibits or blocks the binding of another antibody or antigen-binding fragment thereof. The term also includes competition between two antibodies, in both orientations, i.e., a first antibody that binds and blocks the binding of the second antibody and vice versa. In certain embodiments, the first antibody and the second antibody may bind to the same epitope. Alternatively, the first and second antibodies may bind to different, but overlapping, epitopes, such that the binding of one inhibits or blocks the binding of the second antibody, for example, through steric hindrance. The cross-competition between antibodies can be measured by methods known in the art, for example, by a real-time labelless biolayer interferometry assay.To determine whether a test antibody cross-competes with a reference Ebola virus anti-GP antibody of the invention, the reference antibody is allowed to bind to an Ebola virus GP or peptide under saturation conditions. The ability of the test antibody to bind to the Ebola virus GP is then evaluated. If the test antibody is able to bind to the Ebola virus GP after saturation binding with the reference Ebola virus anti-GP antibody, it can be concluded that the test antibody binds to a different epitope than the reference Ebola virus antibody. Conversely, if the test antibody is unable to bind to the Ebola virus GP after saturation binding with the reference Ebola virus anti-GP antibody, then the test antibody may bind to the same epitope as the epitope bound by the reference Ebola virus anti-GP antibody of the invention.

[00112] The term “substantial identity” or “substantially identical,” when referring to a nucleic acid or fragment thereof, indicates that, when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its chain) Petition 870260058248, dated 06 / 15 / 2026, p. 52 / 143 / 106 supplementary), there is no nucleotide sequence identity in at least about 90%, and more preferably at least about 95%, 96%, 97%, 98% or 99% of the nucleotide bases, as measured by any well-known sequence identity algorithm, such as FASTA, BLAST or GAP, as discussed below. A nucleic acid molecule with substantial identity to a reference nucleic acid molecule may, in certain cases, encode a polypeptide having the same or similar amino acid sequence to the polypeptide encoded by the reference nucleic acid molecule.

[00113] As applied to polypeptides, the term “substantial similarity” or “substantially similar” means that two peptide sequences, when aligned as best as possible, such as by the GAP or BESTFIT programs using standard gap weights, share at least 90% sequence identity, even more preferably at least 95%, 98%, or 99% sequence identity. Preferably, the positions of residues that are not identical differ by conservative amino acid substitutions. A “conservative amino acid substitution” is one in which an amino acid residue is replaced by another amino acid residue with a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein.In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percentage or degree of similarity can be adjusted upward to correct for the conservative nature of the substitution. Methods for making this adjustment are well known to those skilled in the art. See, for example, Pearson (1994) Methods MoI. Biol. 24: 307-331). Examples of groups of amino acids with side chains having similar chemical properties include 1) aliphatic side chains: glycine, alanine, valine, leucine. Petition 870260058248, dated 06 / 15 / 2026, page 53 / 143 / 106 and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartate and glutamate; and 7) sulfur-containing side chains: cysteine ​​and methionine. Preferred conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine. Alternatively, a conservative substitution is any change with a positive value in the PAM250 logarithmic probability matrix described in Gonnet et al. (1992) Science 256: 1443 45. A “moderately conservative” substitution is any change with a non-negative value in the PAM250 logarithmic probability matrix.

[00114] Sequence similarity for polypeptides is typically measured using sequence analysis software. Protein analysis software compares similar sequences using similarity measures attributed to various substitutions, deletions, and other modifications, including conservative amino acid substitutions. For example, the GCG software contains programs such as GAP and BESTFIT that can be used with standard parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild-type protein and a mutant of the same type. See, for example, GCG version 6.1. Polypeptide sequences can also be compared using FASTA with standard or recommended parameters; a program in GCG version 6.1.FASTA (e.g., FASTA2 and FASTA3) provides alignments and percentage sequence identity of regions with the best overlap between query and search sequences (Pearson (2000), supra). Another preferred algorithm when comparing with a sequence. Petition 870260058248, dated 06 / 15 / 2026, page 54 / 143 / 106 of the invention with a database containing a large number of sequences from different organisms is the BLAST computer program, especially BLASTP or TBLASTN, using standard parameters. See, for example, Altschul et al. (1990) J. Mol. Biol. 215: 403-410 and (1997) Nucleic Acids Res. 25:3389-3402.

[00115] The phrase “therapeutically effective amount” means an amount that produces the desired effect for which it is administered. The exact amount will depend on the purpose of the treatment and will be determinable by a person skilled in the art using known techniques (see, for example, Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding).

[00116] As used in this invention, the term “individual” refers to an animal, preferably a mammal, and more preferably a human being who needs improvement, prevention and / or treatment of a disease or disorder, such as a viral infection. The individual may have an Ebola virus infection or be predisposed to developing an Ebola virus infection.Individuals “predisposed to developing an Ebola virus infection” or individuals “who may be at high risk of contracting an Ebola virus infection” include those with compromised immune systems due to autoimmune disease, people receiving immunosuppressive therapy (e.g., after organ transplantation), people afflicted with human immunodeficiency syndrome (HIV) or acquired immunodeficiency syndrome (AIDS), certain forms of anemia that deplete or destroy white blood cells, people receiving radiation or chemotherapy, or people afflicted with an inflammatory disorder. Additionally, extremely young or elderly individuals are at increased risk. Anyone who comes into physical contact or close proximity with an infected animal or human patient, or who is exposed to bodily fluids or tissues, is also at risk. Petition 870260058248, dated 06 / 15 / 2026, page 55 / 143 / 106, of a human or animal patient infected, has a higher risk of developing an Ebola virus infection. Furthermore, a subject is at risk of contracting an Ebola virus infection due to proximity to an outbreak of the disease, for example, an individual residing in a densely populated city or near individuals with confirmed or suspected Ebola virus infections, or choice of employment, for example, a hospital worker, a pharmaceutical researcher, an individual who has visited or plans to visit an area or country known to have or suspected of having an Ebola virus outbreak, or a frequent airline passenger.

[00117] As used in the present invention, the terms “treat,” “treating,” or “treatment” refer to the reduction or improvement in the severity of at least one symptom or indication of Ebola virus infection due to the administration of a therapeutic agent, such as an antibody of the present invention, to an individual in need thereof. The terms include inhibition of disease progression or worsening of infection. The terms also include positive disease prognosis, i.e., the individual may be infection-free or may have reduced or absent viral titers after administration of a therapeutic agent, such as an antibody of the present invention. The therapeutic agent may be administered at a therapeutic dose for the individual.

[00118] The terms “prevents”, “preventing” or “prevention” refer to the inhibition of the manifestation of Ebola virus infection or any symptoms or indications of Ebola virus infection after administration of an antibody of the present invention. The term includes preventing the spread of infection in an individual exposed to the virus or at risk of having Ebola virus infection.

[00119] As used in the present invention, the term "antiviral drug" refers to any anti-infective agent or therapy, whether chemical or biological, that is used for Petition 870260058248, dated 06 / 15 / 2026, p. 56 / 143 / 106 to treat, prevent, or alleviate a viral infection in an individual. For example, in the present invention, an antiviral drug may include, but is not limited to, an antibody to the Ebola virus (in one embodiment, the antibody to the Ebola virus may be different from those described in this document), an Ebola virus vaccine, ZMapp therapy, TKM Ebola (small interfering RNAs targeting viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (phosphorodiamidatomorpholino antisense oligomers targeting the VP24 gene of the Ebola virus), and interferons. In the present invention, the infection to be treated is caused by an Ebola virus. General description

[00120] Ebola virus disease is a serious, often fatal illness caused by filamentous viral particles that are members of the Filoviridae family. There are several known species of the Ebola virus genus that are capable of causing disease in humans. These include Zaire, Sudan, Tai Forest (formerly Côte d'Ivoire), and Bundibugyo. The natural reservoir for the virus is unknown, and to date, there are no approved vaccines or therapies.

[00121] The virus genome consists of a single strand of negative-sense RNA approximately 19 kb in length. Ebola virions contain seven proteins: a surface glycoprotein (GP), a nucleoprotein (NP), four virion structural proteins (VP40, VP35, VP30, and VP24), and an RNA-dependent RNA polymerase (L). (Feldman, et al. (1992) Virus Res. 24, 1-19).

[00122] The only protein present on the surface of the virus is the glycoprotein. Due to RNA editing, transcription of the GP gene results in the synthesis of several GP gene-specific mRNAs that encode viral Gps, including non-structural soluble GP (sGP) and virion surface GP (Volchkova, VA et al., (1998), 250:408 Virology 250:408-414). Both Petition 870260058248, dated 06 / 15 / 2026, p. 57 / 143 / 106 GPs are synthesized as a precursor molecule that is proteolytically cleaved by the cellular protease furin during intracellular processing (Volchkov, VE, et al., (1998), Proc Natl Acad Sci USA 95: 5762-5767). sGP forms dimers, while the cleaved carboxy-terminal fragment is a monomer. The spikes on the viral surface are formed as a trimer of GPi,2 composed of two subunits GP1 and GP2 linked by a disulfide bond (Volchkova, VA et al., (1998), Virology 250:408-414; Falzarano, D. et al., (2006), Chembiochem 7:1605-1611). GP1 is known to mediate viral binding to the host cell, and GP2 is involved in membrane fusion (Sanchez, A. et al., (1996), Proc Natl Acad Sci USA 93:3602-3607; Alazard-Dany, N., et al. (2006), J. Gen. Virol. 87:1247-1257).

[00123] During EBOV infection, significant amounts of soluble glycoproteins (sGP) are released from virus-infected cells. This form of GP has been shown to bind to and sequester virus-neutralizing antibodies directed against the surface or GP of the virion (Dolnik, O. et al., (2004), EMBO J 23:2175-2184). Beyond this antibody blockade, the role of soluble GP in terms of viral replication and / or pathogenicity has not been well defined. More recent studies by Escudero-Perez, et al., have demonstrated that sGP can bind to and activate uninfected dendritic cells and macrophages, and induce the secretion of pro- and anti-inflammatory cytokines. Furthermore, they demonstrated that sGP affects endothelial cell function and may affect vascular permeability. (Escudero-Perez, et al., (2014), PLOS Pathogens, Vol. 10, Issue 11: 1-17).This may explain the dysregulated inflammatory response of the host after infection, and may contribute to the pathogenicity of the virus.

[00124] Passive immunotherapy for the prophylaxis or treatment of infectious diseases has been used for over a century, generally in the form of convalescent human sera containing high titers of Petition 870260058248, dated 06 / 15 / 2026, page 58 / 143 / 106 neutralizing antibodies (Good et al., (1991); Cancer 68: 1415-1421). Nowadays, several purified monoclonal antibodies are currently in preclinical and clinical development for use as antimicrobials (Marasco et al 2007; Nature Biotechnology 25: 1421-1434). Certain antibodies have been described that bind to the Ebola virus glycoprotein. (See, for example, Audet et. al. (2014), Scientific Reports 4:6881; Chen, et.al. (2014), ACS Chem Biol. Oct. 17; 9(10):2263-73; Koellhoffer JF, et.al., (2012), Chembiochem Nov. X., et. al., Nature (2014) October 2; 514(7520): 47-53).

[00125] The inventors have discovered in the present invention fully human antibodies and antigen-binding fragments thereof that specifically bind to the GP of the Ebola virus and modulate the interaction of the Ebola virus with these cells. The anti-GP antibodies of the Ebola virus can bind to the Ebola virus with high affinity. In certain embodiments, the antibodies of the present invention are blocking antibodies, wherein the antibodies can bind to the GP of the Ebola virus and block the binding and / or entry of the virus into host cells. In certain embodiments, the antibodies of the invention can block the binding of the Ebola virus to cells and, as such, they can inhibit or neutralize viral infection of host cells. In certain embodiments, the antibodies of the invention can mediate antibody-dependent cell-mediated cytotoxicity (ADCC) and, as such, they can assist in the destruction of virus-infesting cells.In certain modalities, antibodies can act in both ways; for example, they can neutralize viral infectivity and they can mediate ADCC. In some modalities, antibodies can be useful in treating an individual suffering from an Ebola virus infection. Antibodies, when administered to an individual in need, can reduce infection by a virus, such as the Ebola virus, in that individual. They can be used to decrease viral loads in an individual. They can also be used alone. Petition 870260058248, dated 06 / 15 / 2026, p. 59 / 143 / 106 or as adjuvant therapy with other therapeutic portions or modalities known in the art to treat a viral infection. In certain embodiments, these antibodies can bind to an epitope at the amino terminus of the GP of the Ebola virus. In certain embodiments, these antibodies can bind to an epitope at the carboxyl terminus of the GP of the Ebola virus. Furthermore, the identified antibodies can be used prophylactically (before infection) to protect a mammal against infection or can be used therapeutically (after infection is established) to alleviate a previously established infection or to alleviate at least one symptom associated with the infection.

[00126] The complete amino acid sequence of an Ebola virus GP is shown in GenBank as registry numbers AHX24649.1 and AHX24649.2, and also in SEQ ID NOs: 314 and 315, respectively. GP1 extends from amino acid residues 1 to 501 of the complete GP and GP2 extends from amino acid residues 502 to 676 of the complete GP shown in SEQ ID NO: 314 or SEQ ID NO: 315). The complete EBOV GP, also shown in registry number AHX24649.1, can be coupled to a decahistidine marker, as shown in SEQ ID NO: 318. The soluble GP (sGP) is shown as GenBank accession number AHX24650. 1 and also as SEQ ID NO: 316 (with the signal sequence on) and also as SEQ ID NO: 317 (without the signal sequence, but containing a myc-myc-his marker).

[00127] In certain embodiments, the antibodies of the invention are obtained from mice immunized with a primary immunogen, such as the complete Ebola virus GP or with a recombinant form of the Ebola virus GP or fragments thereof, followed by immunization with a secondary immunogen or with an immunogenically active fragment of the Ebola virus GP. In certain embodiments, the antibodies are obtained from mice immunized with DNA encoding the complete Ebola virus GP (Zaire.2014, Petition 870260058248, dated 06 / 15 / 2026, page 60 / 143 / 106 (see GenBank KJ660346.2; and also SEQ ID NO: 313). The antigen may be a biologically active and / or immunogenic fragment of the Ebola virus GP or DNA encoding the active fragment thereof. The fragment may be derived from any region of the viral GP, including the amino-terminal fragment (e.g., GP1) or the carboxyl-terminal fragment (e.g., GP2). Peptides may be modified to include the addition or substitution of certain residues for labeling or for the purpose of conjugation with carrier molecules such as KLH. For example, a cysteine ​​may be added to either the N-terminal or C-terminal end of a peptide, or a linker sequence may be added to prepare the peptide for conjugation, e.g., KLH for immunization.

[00128] Certain Ebola antiviral antibodies of the present invention are capable of binding to and neutralizing Ebola virus activity, as determined by in vitro or in vivo assays. The ability of the antibodies of the invention to bind to and neutralize Ebola virus activity, and therefore the binding and / or entry of the virus into a host cell, followed by viral infection, can be measured using any standard method known to those skilled in the art, including binding assays or activity assays, as described in the present invention.

[00129] Exemplary and non-limiting in vitro assays for measuring binding activity are illustrated in Example 3 of the present invention. In Example 3, the binding affinity and dissociation constants of anti-GP antibodies to the Ebola virus were determined by Biacore. In Examples 4 and 7, neutralization assays were used to determine the infectivity of various strains of the Ebola virus.

[00130] Specific antibodies to the Ebola virus GP may not contain additional markers or portions, and they may contain an N-terminal or C-terminal marker or portion. In one embodiment, the marker or Petition 870260058248, dated 06 / 15 / 2026, page 61 / 143 / 106 portion is biotin. In a binding assay, the location of a marker (if any) can determine the orientation of the peptide relative to the surface to which the peptide is bound. For example, if a surface is coated with avidin, a peptide containing an N-terminal will be oriented so that the C-terminal portion of the peptide is distal to the surface. In one embodiment, the marker may be a radionuclide, a fluorescent dye, or a marker detectable by magnetic resonance imaging. In certain embodiments, such labeled antibodies can be used in diagnostic assays, including image capture assays. Antigen-binding fragments of antibodies

[00131] Unless specifically indicated otherwise, the term “antibody,” as used in this invention, shall be interpreted to encompass antibody molecules comprising two immunoglobulin heavy chains and two immunoglobulin light chains (i.e., “complete antibody molecules”), as well as the antigen-binding fragments thereof. The terms “antigen-binding portion” of an antibody, “antigen-binding fragment” of an antibody, and the like, as used in the present invention, include any genetically engineered, synthetic, enzymatically obtainable, or naturally occurring polypeptide or glycoprotein that specifically binds to an antigen to form a complex. The terms “antigen-binding fragment” of an antibody or “antibody fragment,” as used in this invention, refer to one or more fragments of an antibody that retain the ability to bind to the Ebola virus.An antibody fragment may include a Fab fragment, an F(ab')2 fragment, an Fv fragment, a dAb fragment, a fragment containing a CDR, or an isolated CDR. In certain embodiments, the term "antigen-binding fragment" refers to a polypeptide fragment. Petition 870260058248, dated 06 / 15 / 2026, pp. 62 / 143 / 106 of a multispecific antigen-binding molecule. Antibody antigen-binding fragments can be derived, for example, from complete antibody molecules using any appropriate standard techniques, such as proteolytic digestion or recombinant genetic manipulation techniques involving the manipulation and expression of the antibody encoding variable DNA and (optionally) constant domains. This DNA is known and / or readily available from, for example, commercial sources, DNA libraries (including, for example, phage-antibody libraries) or it can be synthesized. The DNA can be sequenced and manipulated chemically or by the use of molecular biology techniques, for example, to arrange one or more variable and / or constant domains in an appropriate configuration or to introduce codons, create cysteine ​​residues, modify, add or delete amino acids, etc.

[00132] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv molecules (scFv); (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity-determining region (CDR), such as a CDR3 peptide) or a limited FR3-CDR3-FR4 peptide. Other manipulated molecules, such as domain-specific antibodies, single-domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.).Small modular immunopharmaceuticals (SMIPs) and shark IgNAR variable domains are also encompassed in the expression "antigen-binding fragment," as used in the present invention.

[00133] An antigen-binding fragment of an antibody Petition 870260058248, dated 06 / 15 / 2026, pp. 63 / 143 / 106, will typically comprise at least one variable domain. The variable domain may be of any size or amino acid composition and will generally include at least one CDR, which is adjacent to or in the structure with one or more structure sequences. In antigen-binding fragments with a VH domain associated with a VL domain, the VH and VL domains may be situated relative to each other in any appropriate arrangement. For example, the variable region may be dimeric and contain the dimers Vh-Vh, Vh-Vl, or Vl-Vl. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric Vh or Vl domain.

[00134] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Exemplary and non-limiting configurations of the variable and constant domains that may be found in an antigen-binding fragment of an antibody of the present invention include: (i) Vh-Ch1; (ii) Vh-Ch2; (iii) Vh-Ch3; (iv) Vh-Ch1-Ch2; (v) Vh-Ch1-Ch2-Ch3; (vi) Vh-Ch2-Ch3; (vii) Vh-Cl; (viii) Vl-Ch1; (ix) Vl-Ch2; (x) Vl-Ch3; (xi) Vl-Ch1-Ch2; (xii) Vl-Ch1-Ch2-Ch3; (xiii) Vl-Ch2-Ch3; and (xiv) Vl-Cl. In any variable and constant domain configuration, including any of the exemplary configurations listed above, the variable and constant domains can be directly linked to each other or can be linked by a full or partial hinge or linker region.A hinge region may consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids, resulting in a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a peptide molecule. Furthermore, an antigen-binding fragment of an antibody of the present invention may comprise a homodimer or a heterodimer (or other multimer) in any of the variable and constant domain configurations listed above, in non-covalent association. Petition 870260058248, dated 06 / 15 / 2026, page 64 / 143 / 106 between themselves and / or with one or more monomeric Vh or Vl domains (for example, through disulfide bonds).

[00135] As with complete antibody molecules, antigen-binding fragments can be monospecific or multispecific (e.g., bispecific). A multispecific antigen-binding fragment of an antibody will typically comprise at least two different variable domains, where each variable domain is capable of specifically binding to a separate antigen or to a different epitope on the same antigen. Any multispecific antibody format, including the exemplary bispecific antibody formats described in the present invention, can be adapted for use in the context of an antigen-binding fragment of an antibody of the present invention using routine techniques available in the art. Preparation of human antibodies

[00136] Methods for generating human antibodies in transgenic mice are known in the art. Any of these methods can be used in the context of the present invention to make human antibodies that bind specifically to the Ebola virus GP. An immunogen comprising any of the following can be used to generate antibodies against the Ebola virus. In certain embodiments, the antibodies of the invention are obtained from mice immunized with a native and complete Ebola virus GP (see, for example, GenBank registry numbers AHX24649.1 (SEQ ID NO: 314) and AHX24649.2 (SEQ ID NO: 315)) or with DNA encoding the glycoprotein or a fragment thereof. Alternatively, the Ebola virus GP, or a fragment thereof, can be produced using standard and modified biochemical techniques and used as an immunogen. In one embodiment, the immunogen is a produced Ebola virus GP or a fragment thereof.In certain embodiments of the invention, the antigen may be a commercially available GP of the Ebola virus. Petition 870260058248, dated 06 / 15 / 2026, p. 65 / 143 / 106 In certain embodiments, one or more booster injections may be administered. In certain embodiments, the booster injections may comprise one or more commercially available Ebola virus GPs. In certain embodiments, the immunogen may be a recombinant Ebola virus GP expressed in E. coli or in any other eukaryotic or mammalian cells, such as Chinese hamster ovary (CHO) cells.

[00137] Using VELOCIMMUNE® technology (see, for example, Regeneron Pharmaceuticals US patent 6,596,541, VELOCIMMUNE®) or any other known method for generating monoclonal antibodies, high GP chimeric antibodies against Ebola virus are initially isolated, having a human variable region and a mouse constant region. The VELOCIMMUNE® technology involves generating a transgenic mouse having a genome consisting of human light and heavy chain variable regions operably ligated to endogenous mouse constant region loci such that the mouse produces an antibody comprising a human variable region and a mouse constant region in response to antigenic stimulation. The DNA encoding the antibody's heavy and light chain variable regions is isolated and operably ligated to the DNA encoding the human heavy and light chain constant regions.The DNA is then expressed in a cell capable of expressing the complete human antibody.

[00138] Generally, a VELOCIMMUNE™ mouse is challenged with the antigen of interest, and lymphatic cells (such as B cells) are retrieved from the mice expressing the antibodies. The lymphatic cells can be fused with a myeloma cell line to prepare immortal hybridoma cell lines, and such hybridoma cell lines are tested and screened to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. DNA encoding the variable regions of the heavy chain Petition 870260058248, dated 06 / 15 / 2026, page 66 / 143 / 106 and the light chain can be isolated and ligated to the desirable isotypic constant regions of the heavy chain and the light chain. Such an antibody protein can be produced in a cell, such as in a ChO cell. Alternatively, the DNA encoding the antigen-specific chimeric antibodies or the variable domains of the light and heavy chains can be isolated directly from antigen-specific lymphocytes.

[00139] Initially, high-affinity chimeric antibodies are isolated having a human variable region and a mouse constant region. As in the experimental section below, the antibodies are characterized and selected for desirable characteristics, including affinity, selectivity, epitope, etc. The mouse constant regions are replaced with a desired human constant region to generate the complete human antibody of the invention, for example, wild-type or modified IgG1 or IgG4. Although the selected constant region may vary according to the specific use, the antigen-binding affinity and target specificity characteristics reside in the variable region. Bioequivalents

[00140] The anti-GP antibodies to the Ebola virus and the antibody fragments of the present invention comprise proteins with amino acid sequences that vary from those of the antibodies described, but retain the ability to bind to the Ebola virus. Such variant antibodies and antibody fragments comprise one or more additions, deletions, or substitutions of amino acids compared to the original sequence, but exhibit biological activity that is essentially equivalent to that of the antibodies described. Similarly, the DNA sequences encoding the antibody of the present invention comprise sequences that include one or more additions, deletions, or substitutions of nucleotides compared to the described sequence, but encode an antibody or antibody fragment that is essentially bioequivalent to an antibody. Petition 870260058248, dated 06 / 15 / 2026, p. 67 / 143 / 106 or antibody fragment of the invention.

[00141] Two antigen-binding proteins or antibodies are considered bioequivalent if, for example, they are pharmaceutical equivalents or pharmaceutical alternatives whose rate and extent of absorption do not show a significant difference when administered at the same molar dose under similar experimental conditions, such as single or multiple doses. Some antibodies will be considered pharmaceutical equivalents or alternatives if they are equivalent in the extent of their absorption but not in their rate of absorption and may still be considered bioequivalent because such differences in the rate of absorption are intentional and are reflected in the labeling, are not essential for achieving effective body drug concentrations, for example, in chronic use, and are considered clinically insignificant for the particular drug product studied.

[00142] In one embodiment, two antigen-binding proteins are bioequivalent if there are no clinically significant differences in their safety, purity, or potency.

[00143] In one embodiment, two antigen-binding proteins are bioequivalent if a patient can switch one or more times between the reference product and the biologic product without an expected increase in the risk of adverse effects, including a clinically significant alteration in immunogenicity or decreased efficacy, compared with continued therapy without such switching.

[00144] In one embodiment, two antigen-binding proteins are bioequivalent if both act by a common mechanism or mechanisms of action for the condition or conditions of use, to the extent that such mechanisms are known.

[00145] Bioequivalence can be demonstrated by in vivo and in vitro methods. Bioequivalence measurements include, for example, (a) a test Petition 870260058248, dated 06 / 15 / 2026, p. 68 / 143 / 106 in vivo in humans or other mammals, where the concentration of the antibody or its metabolites is measured in blood, plasma, serum, or other biological fluid as a function of time; (b) an in vitro test that has been correlated with and is reasonably predictive of human in vivo bioavailability data; (c) an in vivo test in humans or other mammals, where the appropriate acute pharmacological effect of the antibody (or its target) is measured as a function of time; and (d) in a well-controlled clinical trial that establishes the safety, efficacy, bioavailability, or bioequivalence of an antibody.

[00146] Bioequivalent variants of the antibodies of the invention can be constructed, for example, by making various substitutions of residues or sequences or by excluding terminal or internal residues or sequences not necessary for biological activity. For example, cysteine ​​residues not essential for biological activity can be excluded or replaced with other amino acids to avoid the formation of unnecessary or incorrect intramolecular disulfide bridges during renaturation. In other contexts, bioequivalent antibodies may include antibody variants comprising amino acid alterations that modify the glycosylation characteristics of the antibodies, for example, mutations that eliminate or remove glycosylation. Ebola antiviral antibodies comprising Fc variants

[00147] According to certain embodiments of the present invention, Ebola antiviral antibodies are provided comprising an Fc domain, which comprises one or more mutations that increase or decrease the binding to the FcRn receptor, for example, at acidic pH, compared to neutral pH. For example, the present invention includes Ebola virus anti-GP antibodies comprising a mutation in the Ch2 or Ch3 region of the Fc domain, wherein the mutation(s) increase(s) the affinity of the Fc domain for FcRn in an acidic environment (for example, in an endosome where the pH Petition 870260058248, dated 06 / 15 / 2026, p. 69 / 143 / 106 varies from about 5.5 to about 6.0). Such mutations can result in an increase in the serum half-life of the antibody when administered to an animal. Non-limiting examples of such Fc modifications include, for example, a modification at position 250 (e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., L / Y / F / W or T), 254 (e.g., S or T) and 256 (e.g., S / R / Q / E / D or T); or a modification to heading 428 / 433 (for example, H / L / R / S / P / Q or K) or 434 (for example, A, W, H, F or Y [N434A, N434W, N434H, N434F or N434Y]); or a modification to heading 250 and / or 428; or a modification to heading 307 or 308 (for example, 308F, V308F) and 434.In one embodiment, the modification comprises a 428L modification (e.g., M428L) and a 434S modification (e.g., N434S); a 428L, 259I modification (e.g., V259I) and a 308F modification (e.g., V308F); a 433K modification (e.g., H433K) and a 434 modification (e.g., 434Y); a 252, 254 and 256 modification (e.g., 252Y, 254T and 256E); a 250Q and 428L modification (e.g., T250Q and M428L); and a 307 or 308 modification (e.g., 308F or 308P). In yet another embodiment, the modification comprises a 265A modification (e.g., D265A) and / or a 297A modification (e.g., N297A).

[00148] For example, the present invention includes Ebola antiviral antibodies comprising an Fc domain, comprising one or more pairs or groups of mutations selected from the group consisting of: 250Q and 248L (e.g., T250Q and M248L); 252Y, 254T and 256E (e.g., M252Y, S254T and T256E); 428L and 434S (e.g., M428L and N434S); 257I and 311I (e.g., P257I and Q311I); 257I and 434H (e.g., P257I and N434H); 376V and 434H (e.g., D376V and N434H); 307A, 380A and 434A (e.g., T307A, E380A and N434A); and 433K and 434F (e.g., H433K and N434F). All possible combinations of the aforementioned Fc domain mutations and other mutations in the antibody variable domains described herein are contemplated within the scope of the present invention. Petition 870260058248, dated 06 / 15 / 2026, page 70 / 143 / 106

[00149] The present invention also includes Ebola antiviral antibodies comprising a chimeric heavy chain (Ch) constant region, wherein the chimeric Ch region comprises segments derived from the Ch regions of more than one immunoglobulin isotype. For example, the antibodies of the invention may comprise a chimeric Ch region comprising part or all of the Ch2 domain derived from a human IgG1, human IgG2, or human IgG4 molecule, combined with part or all of a Ch3 domain derived from a human IgG1, human IgG2, or human IgG4 molecule. According to certain embodiments, the antibodies of the invention comprise a chimeric Ch region having a chimeric hinge region.For example, a chimeric hinge may comprise an amino acid sequence of the “upper hinge” (amino acid residues at positions 216 to 227 according to EU numbering) derived from a hinge region of human IgG1, human IgG2, or human IgG4, combined with a “lower hinge” sequence (amino acid residues at positions 228 to 236 according to EU numbering) derived from a hinge region of human IgG1, human IgG2, or human IgG4. According to certain embodiments, the chimeric hinge region comprises amino acid residues derived from an upper hinge of human IgG1 or human IgG4, and amino acid residues derived from a lower hinge of human IgG2. An antibody comprising a chimeric Ch region, as described in the present invention, in certain embodiments, exhibits modified Fc effector functions without negatively affecting the therapeutic or pharmacokinetic properties of the antibody.(See, for example, U.S. Provisional Application No. 61 / 759,578, filed February 1, 2013). Biological characteristics of antibodies.

[00150] In general, the antibodies of the present invention function by binding to the GP of the Ebola virus. For example, the present invention includes Petition 870260058248, dated 06 / 15 / 2026, page 71 / 143 / 106 antibodies and antibody antigen-binding fragments that bind to the GP of the Ebola virus (e.g., at 25°C or 37°C) with a Kd less than 10-7M, as measured by surface plasmon resonance, for example, using the assay format as described in the present invention. In certain embodiments, antibodies or antigen-binding fragments thereof bind to the GP of the Ebola virus with a KD less than about 10 nM, less than about 5 nM, less than about 1 nM, less than about 500 pM, less than 250 pM, or less than 100 pM, as measured by surface plasmon resonance, for example, using the assay format as described in the present invention or a substantially similar assay.

[00151] The present invention also includes antibodies and antigen-binding fragments thereof that bind the Ebola virus with a dissociative half-life (½) greater than about 3 minutes as measured by surface plasmon resonance at 25°C or greater than about 1 minute as measured by surface plasmon resonance at 37°C, for example, and at least a 3-fold increase in dissociative half-life (½) at pH 5 or pH 6; using an assay format as defined herein or a substantially similar assay.In certain embodiments, the antibodies or antigen-binding fragments of the present invention bind to the Ebola virus with a 0½ greater than about 10 minutes, greater than about 30 minutes, greater than about 60 minutes, greater than about 100 minutes, greater than about 300 minutes, greater than about 500 minutes, greater than about 700 minutes, greater than about 200 minutes, greater than about 400 minutes, greater than about 600 minutes, greater than about 800 minutes, greater than about 900 minutes, or greater than about 1000 minutes as measured by surface plasmon resonance at 25°C or 37°C, for example, using a conforming assay format. Petition 870260058248, dated 06 / 15 / 2026, p. 72 / 143 / 106 defined in this document (for example, in an mAb capture or antigen capture format) or a substantially similar assay.

[00152] The present invention also includes antibodies or antigen-binding fragments thereof that neutralize the infectivity of the Ebola virus to its host cells. In some embodiments, the antibodies exhibit neutralizing potential against Zaire.2014 VLPs, with IC50 values ​​ranging from about 10⁻¹¹M to about 10⁻⁹M. The antibodies of the invention also cross-react with Ebola virus VLPs containing GPs from various EBOV strains, including Zaire.1995, Zaire.2014, Ebola Sudan, Bundibugyo, and Cote d'Ivoire. The antibodies of the invention also mediate ADCC, as shown in Example 5. Furthermore, the antibodies of the invention cross-compete with other antibodies that bind to EBOV GP, as shown in Example 6.

[00153] In one embodiment, the invention provides an isolated recombinant antibody or an antigen-binding fragment thereof that specifically binds to the GP of the Ebola virus, wherein the antibody or its fragment exhibits one or more of the following characteristics: (a) it is a fully human monoclonal antibody; (b) it binds to EBOV or a virus-like particle (VLP) expressing an Ebola virus glycoprotein with a dissociation constant (Kd) less than 10-7M, as measured in a surface plasmon resonance assay; (c) it demonstrates at least a 3-fold increase in dissociative half-life (t / 2) at pH 5 or 6 compared to pH 7.4; (d) it demonstrates neutralization of the Ebola Zaire virus with an IC50 ranging from about 10-11M to about 10-9M; (e) it demonstrates antibody-dependent cellular cytotoxicity of Ebola virus-infected cells; (f) cross-reacts with one or more strains of Ebola virus VLPs selected from the group consisting of Zaire.2014, Zaire.1995, Sudan, Bundibugyo and Cote d'Ivoire; (g) competes cross-country with one. Petition 870260058248, dated 06 / 15 / 2026, p. 73 / 143 / 106 reference antibody, wherein the reference antibody comprises an amino acid sequence of the heavy chain variable region (HCVR) and a light chain variable region (LCVR) selected from the group consisting of any of the HCVR and LCVR amino acid sequences in Table 1.

[00154] The antibodies of the present invention may possess one or more of the aforementioned biological characteristics or any combination thereof. Some of the properties of the antibodies of the invention are summarized below. Other biological characteristics of the antibodies of the present invention will be apparent to a person normally skilled in the art from a review of the present description, including the practical examples in the present invention. mAb Properties mAb IC50 Pseudovirus Neutralization (M) Live Virus Neutralization ADCC sGP Binding H1H17161P Neutralizer, ADCC-, 8,3E-11 sGP- Yes No No H1H17139P No neutralizer, No ADCC+, sGP+ No Yes Yes H1H17203P Neutralizer, 2E-10 ADCC+, sGP- No Yes No Epitope Mapping and Related Technologies

[00155] The present invention includes Ebola antiviral antibodies that interact with one or more amino acids found in the GP of the Ebola virus. The epitope to which the antibodies bind may consist of a single contiguous sequence of 3 or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) amino acids located in the GP molecule of the Ebola virus (e.g., a linear epitope in a domain). Alternatively, the epitope may consist of a plurality of non-contiguous amino acids (or amino acid sequences) located in the GP of the Ebola virus (e.g., a conformational epitope).

[00156] Several techniques known to people normally versed in the art can be used to determine whether an antibody “interacts with one or more amino acids” in a polypeptide or protein. Petition 870260058248, dated 06 / 15 / 2026, page 74 / 143 / 106 Exemplary techniques include, for example, routine cross-blocking assays, such as those described in Antibodies, Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harbor, NY). Other methods include alanine scan mutational analysis, peptide blot analysis (Reineke (2004) Methods Mol. Biol. 248: 443-63), peptide cleavage analysis crystallography studies, and NMR analyses. In addition, methods such as epitope excision, epitope extraction, and chemical modification of antigens can be used (Tomer (2000) Prot Sci. 9: 487-496). Another method that can be used to identify the amino acids in a polypeptide with which an antibody interacts is hydrogen / deuterium exchange detected by mass spectrometry. In general terms, the hydrogen / deuterium exchange method involves labeling the protein of interest with deuterium, followed by antibody binding to the deuterium-labeled protein.Next, the protein / antibody complex is transferred to water, and the exchangeable protons in the amino acids that are protected by the antibody complex undergo reverse deuterium-hydrogen exchange at a slower rate than the exchangeable protons in the amino acids that are not part of the interface. As a result, the amino acids that are part of the protein / antibody interface can retain deuterium and therefore have a relatively higher mass compared to the amino acids not included in the interface. After antibody dissociation, the target protein is subjected to protease cleavage and mass spectrometry analysis, thus revealing the deuterium-labeled residues that correspond to the specific amino acids with which the antibody interacts. See, for example, Ehring (1999) Analytical Biochemistry 267: 252-259; Engen and Smith (2001) Anal. Chem. 73: 256A-265A.

[00157] The term “epitope” refers to a site on an antigen to which B and / or T cells respond. B cell epitopes can be formed from contiguous amino acids or from non-contiguous amino acids. Petition 870260058248, dated 06 / 15 / 2026, p. 75 / 143 / 106 juxtaposed by the tertiary fold of a protein. Epitopes formed from contiguous amino acids are normally retained upon exposure to denaturing solvents, while epitopes formed by tertiary folds are typically lost upon treatment with denaturing solvents. An epitope typically includes at least 3, and more commonly, at least 5 or 8 to 10 amino acids in a single spatial conformation.

[00158] Modification-Assisted Profiling (MAP), also known as antigen structure-based antibody profiling (ASAP), is a method that classifies a large number of monoclonal antibodies (mAbs) directed against the same antigen according to the similarities in the binding profile of each antibody to chemically or enzymatically modified antigen surfaces (see US patent 2004 / 0101920). Each category may reflect a unique epitope, either distinctly different from or partially overlapping the epitope represented by another category. This technology allows for the rapid filtering of genetically identical antibodies, so that characterization can be focused on genetically distinct antibodies. When applied to hybridoma screening, MAP can facilitate the identification of rare hybridoma clones that produce mAbs with the desired characteristics.MAP can be used to classify the antibodies of the invention into groups of different antibody-binding epitopes.

[00159] In certain embodiments, Ebola virus antibodies or antigen-binding fragments thereof bind to an epitope in any of the regions exemplified in the Ebola virus GP, in natural or recombinantly produced form, or to a fragment thereof.

[00160] The present invention includes anti-GP antibodies to the Ebola virus that bind to the same epitope or a portion of the epitope. Similarly, the present invention also includes anti-GP antibodies to the Ebola virus that compete for the binding of the Ebola virus GP or a fragment thereof. Petition 870260058248, dated 06 / 15 / 2026, page 76 / 143 / 106 with any of the exemplary specific antibodies described in the present invention. For example, the present invention includes anti-GPP antibodies to the Ebola virus that cross-compete for binding to the Ebola virus with one or more antibodies obtained from those antibodies described in Tables 1 and 2.

[00161] A person can easily determine whether an antibody binds to the same epitope as, or competes for binding with, a reference Ebola virus anti-GP antibody using routine methods known in the art. For example, to determine whether a test antibody binds to the same epitope as a reference Ebola virus anti-GP antibody of the invention, the reference antibody is allowed to bind to an Ebola virus GP or peptide under saturation conditions. The ability of a test antibody to bind to the Ebola virus GP is then evaluated. If the test antibody is able to bind to the Ebola virus GP after saturation binding with the reference Ebola virus anti-GP antibody, it can be concluded that the test antibody binds to a different epitope than the reference Ebola virus antibody.On the other hand, if the test antibody is unable to bind to the GP of the Ebola virus after saturation binding with the reference anti-GP antibody of the Ebola virus, then the test antibody may bind to the same epitope as the epitope bound by the reference anti-GP antibody of the Ebola virus of the invention.

[00162] To determine if an antibody competes for binding with an antibody from the Ebola virus GP, the binding methodology described above is performed in two orientations: in a first orientation, the reference antibody is bound to an Ebola virus GP under saturation conditions, followed by evaluation of the binding of the test antibody to the Ebola virus GP. In a second orientation, the test antibody is allowed to bind to an Ebola virus GP under saturation conditions, followed by evaluation of the binding of the reference antibody to the Ebola virus GP. If, in both orientations, Petition 870260058248, dated 06 / 15 / 2026, p. 77 / 143 / 106, only if the first antibody (saturating antibody) is able to bind to the GP of the Ebola virus, then it is concluded that the test antibody and the reference antibody compete for binding to the GP of the Ebola virus. As will be observed by a person skilled in the art, an antibody that competes for binding with a reference antibody cannot necessarily bind to the identical epitope as the reference antibody, but may sterically block the binding of the reference antibody by binding to an overlapping or adjacent epitope.

[00163] Two antibodies bind to an identical or overlapping epitope if each competitively inhibits (blocks) the binding of the other antigen. That is, a 1, 5, 10, 20, or 100-fold excess of one antibody inhibits the binding of the other by at least 50%, but preferably 75%, 90%, or even 99%, as measured in a competitive binding assay (see, for example, Junghans et al., Cancer Res. 1990 50:1495-1502). Alternatively, two antibodies have the same epitope if essentially all amino acid mutations in the antigen that reduce or eliminate the binding of one antibody reduce or eliminate the binding of the other. Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate the binding of one antibody reduce or eliminate the binding of the other.

[00164] Additional routine experimentation (e.g., peptide mutation and binding analyses) can then be performed to confirm whether the observed lack of binding of the test antibody is in fact due to binding to the same epitope as the reference antibody or whether steric blocking (or another phenomenon) is responsible for the observed lack of binding. Experiments of this type can be performed using ELISA, RIA, surface plasmon resonance, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art. Immunoconjugates Petition 870260058248, dated 06 / 15 / 2026, page 78 / 143 / 106

[00165] The invention comprises a human monoclonal antibody of the Ebola virus GP conjugated with a therapeutic moiety (“immunoconjugate”, as an antiviral drug to treat Ebola virus infection. As used in the present invention, the term “immunoconjugate” refers to an antibody that is chemically or biologically linked to a radioactive agent, a cytokine, an interferon, a target or reporter moiety, an enzyme, a peptide or protein, or a therapeutic agent. The antibody may be linked to the radioactive agent, cytokines, interferon, a target or reporter moiety, enzyme, therapeutic agent, or peptide at any location along the molecule, provided that it is capable of binding to its target. Examples of immunoconjugates include antibody-drug conjugates and antibody-toxin fusion proteins.In certain embodiments, the antibody can be conjugated with an agent specific to a virally infected cell. The type of therapeutic moiety that can be conjugated with the Ebola antiviral antibody will depend on the condition being treated and the desired therapeutic effect to be achieved. Examples of suitable agents for forming immunoconjugates are known in the art; see, for example, WO 05 / 103081. Multispecific antibodies

[00166] The antibodies of the present invention may be monospecific, bispecific, or multispecific. Multispecific antibodies may be specific for different epitopes of a target polypeptide or may contain antigen-binding domains specific for more than one target polypeptide. See, for example, Tutt et al., 1991, J. Immunol. 147:60-69; Kufer et al., 2004, Trends Biotechnol. 22: 238-244.

[00167] Any of the multispecific antigen-binding molecules of the invention, or variants thereof, may be constructed. Petition 870260058248, dated 06 / 15 / 2026, page 79 / 143 / 106 using standard molecular biology techniques (e.g., recombinant DNA technology and protein expression technology), such as will be known to a person normally skilled in the art.

[00168] In some embodiments, antibodies specific to the Ebola virus are generated in a bispecific (“bispecific”) format in which variable regions that bind to distinct domains of the Ebola virus are joined together to confer dual domain specificity within a single binding molecule. Properly designed bispecifics can increase the effectiveness of Ebola virus protein inhibition by increasing both specificity and binding avidity. Variable regions with specificity for individual domains (e.g., N-terminal domain segments) or that can bind to different regions within a domain are paired in a structure that allows each region to simultaneously bind to separate epitopes or to different regions within a domain.In one example, bispecific heavy chain variable (VH) regions of a ligand with specificity for one domain are recombined with light chain variable (VL) regions of a series of ligands with specificity for a second domain to identify non-cognate VL partners that can be paired with an original VH without disrupting the original specificity for that VH. In this way, a single VL segment (e.g., VL1) can be combined with two different VH domains (e.g., Vh1 and Vh2) to generate a bispecific composed of two linking “arms” (VH1-VL1 and VH2-VL1). The use of a single VL segment reduces the complexity of the system and thus simplifies and increases the efficiency of the cloning, expression, and purification processes used to generate the bispecifics (see, for example, USSN13 / 022759 and US2010 / 0331527).

[00169] Alternatively, antibodies that bind to more than one domain and a second target, such as, but not limited to, for example, a Petition 870260058248, dated 06 / 15 / 2026, page 80 / 143 / 106. According to a different Ebola antiviral antibody, they can be prepared in a bispecific format using the techniques described in the present invention or other techniques known to those skilled in the art. The variable regions of the antibody that bind to distinct regions can be joined with variable regions that bind to relevant sites, for example, in the Ebola virus, to confer dual antigen specificity in a single binding molecule. Properly designed bispecifics of this nature serve a dual function. Variable regions with specificity for the extracellular domain are combined with a variable region with specificity outside the extracellular domain, and are paired in a structure that allows each variable region to bind to separate antigens.

[00170] An exemplary bispecific antibody format that can be used in the context of the present invention involves the use of a first immunoglobulin (Ig) Ch3 domain and a second Ig Ch3 domain, wherein the first and second Ig CH3 domains differ from each other by at least one amino acid, and wherein at least one amino acid difference reduces the binding of the bispecific antibody to protein A, compared with a bispecific antibody without the amino acid difference. In one embodiment, the first Ig CH3 domain binds to protein A and the second Ig CH3 domain contains a mutation that reduces or eliminates protein A binding, such as an H95R modification (for exon numbering IMGT; H435R for exon numbering EU). The second Ch3 may further comprise a Y96F modification (for IMGT; Y436F for EU).Other modifications that can be found in the second Ch3 include: D16E, L18M, N44S, K52N, V57M, and V82I (by IMGT; D356E, L358M, N384S, K392N, V397M, and V422I by EU) in the case of IgG1 antibodies; N44S, K52N, and V82I (IMGT; N384S, K392N, and V422I by EU) in the case of IgG2 antibodies; and Q15R, N44S, K52N, V57M, R69K, E79Q, and V82I (by IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q, and V422I by EU) in the case of IgG4 antibodies. Variations in... Petition 870260058248, dated 06 / 15 / 2026, page 81 / 143 / 106, the bispecific antibody format described above is contemplated within the scope of the present invention.

[00171] Other exemplary bispecific formats that may be used in the context of the present invention include, without limitation, for example, scFv-based or bispecific diabodies, IgG-scFv fusions, dual variable domain (DVD)-Ig, quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, CrossFab, (SEED)body, leucine zipper, Duobody, IgG1 / IgG2, dual-action Fab (DAF)-IgG and bispecific Mab2 formats (see, for example, Klein et al. 2012, mAbs 4:6, 1-11, and the references cited therein, for a review of the above formats). Bispecific antibodies can also be constructed using peptide / nucleic acid conjugation, for example, where artificial amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates that then self-assemble into multimeric complexes with defined composition, valence, and geometry.(See, for example, Kazane et al., J. Am. Chem. Soc. [Epub: December 4, 2012]). Therapeutic Administration and Formulations

[00172] The invention provides therapeutic compositions comprising anti-GP antibodies to the Ebola virus or antigen-binding fragments thereof of the present invention. Therapeutic compositions according to the invention will be administered with suitable vehicles, excipients and other agents that are incorporated into the formulations to provide improved transfer, release, tolerance and the like. A multitude of suitable formulations can be found in the formulary known to all pharmaceutical chemists: Remington Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, Petition 870260058248, dated 06 / 15 / 2026, page 82 / 143 / 106 lipid-containing vesicles (cationic or anionic) (such as LIPOFECTIN™), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, carbowax emulsions (polyethylene glycols of various molecular weights), semi-solid gels and semi-solid mixtures containing carbowax. See also Powell et al. “Compendium of excipients for parenteral formulations” PDA (1998) J Pharm Sci Technol 52:238-311.

[00173] The antibody dose may vary depending on the age and size of the individual to be administered, the target disease, conditions, route of administration, and the like. When an antibody of the present invention is used in the treatment of a disease or disorder in an adult patient or for the prevention of such a disease, it is advantageous to administer the antibody of the present invention normally in a single dose of about 0.1 to about 60 mg / kg of body weight, more preferably from about 5 to about 60, from about 10 to about 50, or from about 20 to about 50 mg / kg of body weight. Depending on the severity of the condition, the frequency and duration of treatment may be adjusted.In certain embodiments, the antibody or antigen-binding fragment of the invention may be administered as an initial dose of at least about 0.1 mg to about 800 mg, from about 1 mg to about 500 mg, from about 5 mg to about 300 mg, or from about 10 mg to about 200 mg, up to about 100 mg or up to about 50 mg. In certain embodiments, the initial dose may be followed by the administration of a second or a plurality of subsequent doses of the antibody or antigen-binding fragment in an amount that may be approximately equal to or less than that of the initial dose, the subsequent doses being separated by at least 1 day to 3 days; at least one week; at least 2 weeks; at least 3 weeks; at least 4 weeks; at least 5 weeks; at least 6 weeks; at least 7 weeks; at least 8 weeks; at least 9 weeks; at least 10 weeks; at least 12 weeks; or at least 14. Petition 870260058248, dated 06 / 15 / 2026, p. 83 / 143 / 106 weeks.

[00174] Various delivery systems are known and can be used to administer the pharmaceutical composition of the invention, for example, encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see, for example, Wu et al. (1987) J. Biol. Chem. 262: 4429-4432). Methods of delivery include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition can be administered by any convenient route, for example, by bulk injection or infusion, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and can be administered together with other biologically active agents. Administration can be local or systemic.The pharmaceutical composition can also be administered in a vesicle, in particular in a liposome (see, for example, Langer (1990) Science 249:1527-1533).

[00175] The use of nanoparticles to deliver the antibodies of the present invention is also contemplated in the present invention. Antibody-conjugated nanoparticles can be used for both therapeutic and diagnostic applications. Antibody-conjugated nanoparticles and methods of preparation and use are described in detail by Arruebo, M., et al. 2009 (“Antibody-conjugated nanoparticles for biomedical applications” in J. Nanomat. Volume 2009, article ID 439389, 24 pages, doi: 10.1155 / 2009 / 439389). Nanoparticles can be developed and conjugated with the antibodies contained in pharmaceutical compositions to target virally infected cells. Nanoparticles for drug delivery have also been described, for example, in documents US 8257740 or US 8246995.

[00176] In certain situations, the pharmaceutical composition may Petition 870260058248, dated 06 / 15 / 2026, page 84 / 143 / 106 to be administered in a controlled-release system. In one embodiment, a pump may be used. In another embodiment, polymeric materials may be used. In yet another embodiment, a controlled-release system may be placed close to the target of the composition, requiring only a fraction of the systemic dose.

[00177] Injectable preparations may include dosage forms for intravenous, subcutaneous, intracutaneous, intracranial, intraperitoneal, and intramuscular injections, drip infusions, etc. These injectable preparations may be prepared by publicly known methods. For example, injectable preparations may be prepared, for instance, by dissolving, suspending, or emulsifying the antibody or its salt described above in a sterile aqueous medium or in an oily medium conventionally used for injections. As an aqueous medium for injections, there is, for example, physiological saline, an isotonic solution containing glucose and other adjuvants, etc., which can be used together with a suitable solubilizing agent, such as an alcohol (e.g., ethanol), a polyalcohol (e.g., propylene glycol, polyethylene glycol), a non-ionic surfactant [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) hydrogenated castor oil adduct)], etc.As an oily medium, sesame oil, soybean oil, etc., are used, which can be used together with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. The injection thus prepared is preferably filled into an appropriate ampoule.

[00178] A pharmaceutical composition of the present invention can be administered subcutaneously or intravenously using a standard syringe and needle. Furthermore, with respect to subcutaneous administration, a pen-type administration device has applications in administering a pharmaceutical composition of the present invention. Such a pen-type administration device can be reusable or disposable. A device Petition 870260058248, dated 06 / 15 / 2026, page 85 / 143 / 106. A reusable pen-type administration device generally uses a replaceable cartridge containing a pharmaceutical composition. Once all the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be readily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen-type administration device can then be reused. In a disposable pen-type administration device, there is no replaceable cartridge. Instead, the pen-type administration device is pre-loaded with the pharmaceutical composition held in a reservoir inside the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[00179] Various reusable pen-type administration devices and autoinjectors have applications in the subcutaneous administration of a pharmaceutical composition of the present invention. Examples include, but are certainly not limited to, AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Burghdorf, Switzerland), HUMALOG MIX 75 / 25™ pen, HUMALOG™ pen, HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™ and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), to name a few.Examples of disposable pen-type delivery devices with applications in the subcutaneous administration of a pharmaceutical composition of the present invention include, but are certainly not limited to, the SOLOSTAR™ pen (Sanofi-Aventis), FLEXPEN™ (Novo Nordisk) and KWIKPEN™ (Eli Lilly), the SURECLICK™ autoinjector (Amgen, Thousand Oaks, CA), the PENLET™ (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, LP), to name just a few. Examples of disposable pen delivery. Petition 870260058248, dated 06 / 15 / 2026, page 86 / 143 / 106 devices that have applications in subcutaneous delivery of a pharmaceutical composition of the present invention include, but are certainly not limited to, the SOLOSTAR™ pen (Sanofi-Aventis), the FLEXPEN™ (Novo Nordisk) and the KWIKPEN™ (Eli Lilly), the SURECLICK™ autoinjector (Amgen, Thousand Oaks, CA), the PENLET™ (Haselmeier, Stuttgart, Germany), the EPINEPHRINE (Dey, LP) and the HUMIRA™ pen (Abbott Labs, Abbott Park, IL), to name just a few.

[00180] Advantageously, the pharmaceutical compositions for oral or parenteral use described above are prepared in single-dose dosage forms suitable to accommodate a dose of the active ingredients. Such single-dose dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of antibody contained is generally from about 5 to about 500 mg per single-dose dosage form; especially in the injection form, it is preferable that the antibody be contained in about 5 to about 100 mg and in about 10 to about 250 mg for the other dosage forms. Therapeutic uses of antibodies

[00181] The antibodies of the present invention are useful for the treatment and / or prevention of a disease, disorder or condition associated with Ebola virus infection and / or for improving at least one symptom associated with such disease, disorder or condition.

[00182] In certain embodiments, the antibodies of the invention are useful for treating individuals suffering from severe and acute respiratory infection caused by the Ebola virus. In some embodiments, the antibodies of the invention are useful for decreasing viral titers or for decreasing the viral load in the host. In one embodiment, an antibody or an antigen-binding fragment thereof of the invention can be administered at a therapeutic dose to a patient with Ebola virus infection. Petition 870260058248, dated 06 / 15 / 2026, page 87 / 143 / 106

[00183] One or more antibodies of the present invention can be administered to alleviate, prevent, or reduce the severity of one or more of the symptoms or conditions of disease or disorder. The antibodies can be used to alleviate or reduce the severity of at least one symptom of Ebola virus infection, including, but not limited to, fever, headache, fatigue, loss of appetite, myalgia, diarrhea, vomiting, abdominal pain, dehydration, and unexplained bleeding.

[00184] The present invention also contemplates the use of one or more antibodies of the present invention prophylactically in individuals at risk of developing an Ebola virus infection, such as an immunocompromised individual, a healthcare worker, a person suspected of having been exposed to a person infected with the Ebola virus, a person who comes into physical contact or close physical proximity with an infected individual, a hospital worker, a pharmaceutical researcher, maintenance staff responsible for cleaning the facilities of a hospital or institution where a patient with Ebola has been treated, individuals who have visited or are planning to visit an area or country known to have or suspected of having an Ebola virus outbreak, or a frequent airline passenger.

[00185] In a further embodiment of the invention, the present antibodies are used for the preparation of a pharmaceutical composition for the treatment of patients suffering from an Ebola virus infection. In another embodiment of the invention, the present antibodies are used as adjuvant therapy with any other agent or any other therapy known to those skilled in the art to be useful for treating or attenuating an Ebola virus infection. Combination therapies

[00186] Combination therapies may include an antiGP antibody to the Ebola virus of the invention and any additional therapeutic agent that Petition 870260058248, dated 06 / 15 / 2026, p. 88 / 143 / 106, can be advantageously combined with an antibody of the invention or with a biologically active fragment of an antibody of the invention. The antibodies of the present invention can be combined synergistically with one or more drugs or agents used to treat Ebola virus infection.

[00187] For example, exemplary agents for the treatment of a viral infection may include, for example, antiviral drugs, an anti-inflammatory drug (such as corticosteroids and nonsteroidal anti-inflammatory drugs), a different antibody to the Ebola virus, a vaccine for the Ebola virus, Zmapp therapy, TKM Ebola (small interfering RNAs that target viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (phosphorodiamidatomorpholino antisense oligomers that target the VP24 gene of the Ebola virus), interferons, or any other palliative therapy to treat an Ebola virus infection.

[00188] In some embodiments, the antibodies of the invention can be combined with a second therapeutic agent to reduce the viral load in a patient with an Ebola virus infection or to improve one or more symptoms of the infection.

[00189] In certain embodiments, the second therapeutic agent is another different antibody or antibody cocktail specific to the GP of the Ebola virus, wherein the different antibody or antibodies in the cocktail may or may not bind to the same epitope or to an overlapping epitope, such as an antibody of the present invention. In certain embodiments, the second therapeutic agent is an antibody to a different Ebola virus protein. The second antibody may be specific to one or more different Ebola virus proteins from different strains of the virus. The present invention contemplates the use of a combination (“cocktail”) of the antibodies of the invention with neutralizing or inhibitory activity against the Ebola virus. In some embodiments, non-competing antibodies may be combined and administered to an individual in need to reduce Petition 870260058248, dated 06 / 15 / 2026, p. 89 / 143 / 106, regarding the ability of the Ebola virus to escape due to mutation. In some embodiments, the antibodies comprising the combination bind to distinct non-overlapping epitopes in GP. The antibodies comprising the combination can block virus binding and / or entry into and / or fusion with host cells. The antibodies can interact with GP from a selected EBOV strain from the Zaire, Sudan, Bundibugyo, or Cote d'Ivoire types and, when used alone or in combination with any one or more of the agents noted above, can neutralize any one or more of the noted Ebola virus strains.

[00190] Also contemplated herein is the use of a combination of anti-GP antibodies to the Ebola virus of the present invention, wherein the combination comprises one or more antibodies that do not cross-compete. In certain embodiments, the combination includes a cocktail comprising a mixture of at least three antibodies of the invention. The antibodies in the cocktail may differ in terms of their ability to neutralize the virus or virus-infected cells, or in terms of their ability to mediate antibody-dependent cellular cytotoxicity (ADCC), or in terms of their ability to bind to the soluble glycoprotein of EBOV (sGP).

[00191] As used in the present invention, the term “in combination with” means that the additional therapeutically active component(s) may be administered before, at the same time as, or after the administration of at least one Ebola virus anti-GP antibody of the invention or a cocktail comprising one or more of the antibodies of the present invention. The term “in combination with” also includes the sequential or concomitant administration of an Ebola virus anti-GP antibody and a second therapeutic agent.

[00192] The additional therapeutically active component(s) may be administered to an individual prior to the administration of an anti-GP antibody to the Ebola virus of the present invention. For example, one might consider Petition 870260058248, dated 06 / 15 / 2026, page 90 / 143 / 106, a first component to be administered “before” a second component if the first component is administered 1 week before, 72 hours before, 60 hours before, 48 hours before, 36 hours before, 24 hours before, 12 hours before, 6 hours before, 5 hours before, 4 hours before, 3 hours before, 2 hours before, 1 hour before, 30 minutes before, 15 minutes before, 10 minutes before, 5 minutes before, or less than 1 minute before the administration of the second component. In other embodiments, the additional therapeutically active component(s) may be administered to an individual before the administration of an anti-GP antibody to the Ebola virus of the present invention.For example, a first component may be considered to be administered “after” a second component if the first component is administered 1 minute later, 5 minutes later, 10 minutes later, 15 minutes later, 30 minutes later, 1 hour later, 2 hours later, 3 hours later, 4 hours later, 5 hours later, 6 hours later, 12 hours later, 24 hours later, 36 hours later, 48 hours later, 60 hours later, or 72 hours after the administration of the second component. In still other embodiments, the additional therapeutically active component(s) may be administered to an individual at the same time as the administration of an anti-GP antibody to the Ebola virus of the present invention."Simultaneous" administration, for the purposes of the present invention, includes, for example, the administration of an anti-GP antibody to the Ebola virus and an additional therapeutically active component to an individual in a single dose form or in separate dose forms administered to the individual within about 30 minutes or less of each other. If administered in distinct dose forms, each dose form may be administered via the same route (e.g., both the anti-GP antibody to the Ebola virus and the additional therapeutically active component may be administered intravenously, etc.); alternatively, each dose form may be administered via a different route (e.g., the...). Petition 870260058248, dated 06 / 15 / 2026, pp. 91 / 143 / 106 (Ebola virus anti-GP antibody can be administered intravenously, and the additional therapeutically active component can be administered orally). In any case, administration of the components in a single dosage form, in separate dosage forms via the same route, or in separate dosage forms via different routes is considered “simultaneous administration” for the purposes of this description. For the purposes of this description, administration of an Ebola virus anti-GP antibody “before,” “simultaneously with,” or “after” (as those terms are defined in the present invention above) the administration of an additional therapeutically active component is considered administration of an Ebola virus anti-GP antibody “in combination with” an additional therapeutically active component.

[00193] The present invention includes pharmaceutical compositions in which an anti-GP antibody to the Ebola virus of the present invention is co-formulated with one or more of the additional therapeutically active component(s), as described in another section of the present invention. Management systems

[00194] According to certain embodiments, a single dose of the Ebola virus anti-GP antibody of the invention (or a pharmaceutical composition comprising a combination of an Ebola virus anti-GP antibody and any of the therapeutically active agents mentioned in the present invention) can be administered to an individual in need thereof. According to certain embodiments of the present invention, multiple doses of an Ebola virus anti-GP antibody (or of a pharmaceutical composition comprising a combination of an Ebola virus anti-GP antibody and any of the therapeutically active agents mentioned herein) can be administered to an individual over a defined period of time. The methods according to this aspect of the invention comprise sequentially administering to an individual multiple doses of Petition 870260058248, dated 06 / 15 / 2026, p. 92 / 143 / 106, describes an anti-GP antibody to the Ebola virus of the invention. As used in the present invention, "sequential administration" means that each dose of anti-GP antibody to the Ebola virus is administered to the individual at a different point in time, for example, on different days, separated by a predetermined interval (e.g., hours, days, weeks, or months). The present invention includes methods comprising sequentially administering to the patient a single initial dose of an anti-GP antibody to the Ebola virus, followed by one or more secondary doses of the anti-GP antibody to the Ebola virus, and optionally followed by one or more tertiary doses of the anti-GP antibody to the Ebola virus.

[00195] The terms “initial dose”, “secondary doses” and “tertiary doses” refer to the temporal sequence of administration of the Ebola virus anti-GP antibody of the invention. Thus, the “initial dose” is the dose administered at the beginning of the treatment regimen (also referred to as the “baseline dose”); the “secondary doses” are the doses administered after the initial dose; and the “tertiary” doses are the doses administered after the secondary doses. All initial, secondary and tertiary doses may contain the same amount of Ebola virus anti-GP antibody, but in general, they may differ from each other in terms of frequency of administration. In certain embodiments, however, the amounts of Ebola virus anti-GP antibody contained in the initial, secondary and / or tertiary doses vary from each other (e.g., adjusted upwards or downwards, as the case may be) during the course of treatment.In certain modalities, two or more doses (e.g., 2, 3, 4, or 5) are administered at the start of the treatment regimen as "loading doses," followed by subsequent doses that are administered less frequently (e.g., "maintenance doses").

[00196] In certain exemplary embodiments of the present invention, each secondary or tertiary dose is administered from 1 to 48 hours (for example, 1, Petition 870260058248, dated 06 / 15 / 2026, pp. 93 / 143 / 106 1½ 2, 2½ 3, 3½ 4, 4½ 5, 5½ 6, 6½ 7, 7½ 8, 8½ 9, 9½ 10, 10½ 11, 11½ 12, 12½ 13, 13½ 14, 14½ 15, 15½ 16, 16½ 17, 17½ ​​18, 18½ 19, 19½ 20, 20½ 21, 21½ 22, 22½ 23, 23½ 24, 24½ 25, 25½ 26, 26½ or more) after the immediately preceding dose. The phrase "the immediately preceding dose," as used in the present invention, means, in a sequence of multiple administrations, the dose of the anti-GP antibody to the Ebola virus that is administered to a patient before the administration of the next dose in the sequence without intermediate doses.

[00197] The methods according to this aspect of the invention may comprise administering to a patient any number of secondary and / or tertiary doses of an anti-GP antibody to the Ebola virus. For example, in certain embodiments, only a single secondary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) secondary doses are administered to the patient. Similarly, in certain embodiments, only a single tertiary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) tertiary doses are administered to the patient.

[00198] In certain embodiments of the invention, the frequency at which secondary and / or tertiary doses are administered to the patient may vary throughout the treatment regimen. The frequency of administration may also be adjusted during treatment by a physician, depending on the patient's needs after clinical examination. Diagnostic uses of antibodies

[00199] The anti-GP antibodies to the Ebola virus of the present invention can be used to detect and / or measure the Ebola virus in a sample, for example, for diagnostic purposes. Some embodiments contemplate the use of one or more antibodies of the present invention in assays to detect a disease or disorder, such as a viral infection. Diagnostic assays Petition 870260058248, dated 06 / 15 / 2026, page 94 / 143 / 106 examples for the Ebola virus may include, for example, contacting a sample obtained from a patient with an anti-GP antibody of the Ebola virus of the invention, wherein the anti-GP antibody of the Ebola virus is labeled with a detectable marker or reporter molecule or is used as a capture ligand to selectively isolate the Ebola virus from patient samples. Alternatively, an unlabeled anti-GP antibody of the Ebola virus may be used in diagnostic applications in combination with a secondary antibody, which is itself detectably labeled. The detectable marker or reporter molecule may be a radioisotope, such as 3H,14C,32P,35S or 125I; a fluorescent or chemiluminescent moiety, such as fluorescein isothiocyanate or rhodamine; or an enzyme such as alkaline phosphatase, β-galactosidase, horseradish peroxidase, or luciferase.Specific exemplary assays that can be used to detect or measure the Ebola virus in a sample include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and fluorescence-activated cell separation (FACS).

[00200] Samples that can be used in diagnostic assays for the Ebola virus, according to the present invention, include any tissue or fluid sample obtained from a patient that contains detectable quantities of Ebola virus or fragments thereof, under normal or pathological conditions. Generally, the levels of Ebola virus in a given sample obtained from a healthy patient (e.g., a patient not affected by an Ebola virus-associated illness) will be measured to initially establish a baseline or standard level of Ebola virus. This initial level of Ebola virus can then be compared with the levels of Ebola virus measured in samples from individuals suspected of having an Ebola virus-associated condition or symptoms associated with that condition.

[00201] Specific antibodies for the Ebola virus may not contain Petition 870260058248, dated 06 / 15 / 2026, page 95 / 143 / 106 additional markers or portions, or they may contain an N-terminal or C-terminal marker or portion. In one embodiment, the marker or portion is biotin. In a binding assay, the location of a marker (if any) can determine the orientation of the peptide relative to the surface to which the peptide is bound. For example, if a surface is coated with avidin, a peptide containing an N-terminal will be oriented so that the C-terminal portion of the peptide is distal to the surface. EXAMPLES

[00202] The following examples are presented to provide, to persons normally skilled in the art, a full description and account of how to make and use the methods and compositions of the invention, and are not intended to limit the scope of what the inventors consider to be their invention. Efforts have been made to ensure accuracy with respect to the numbers used (e.g., quantities, temperature, etc.) but some errors and deviations should be taken into account. Unless otherwise indicated, parts are parts by weight, molecular weight is the average molecular weight, temperature is in degrees Celsius, ambient temperature is approximately 25°C and pressure is atmospheric pressure or close thereto. Example 1: Generation of human antibodies to the Ebola virus.

[00203] Human antibodies to Ebola virus were generated in a mouse, comprising the variable kappa light chain and heavy chain regions of human immunoglobulin. In one embodiment, human antibodies to Ebola virus were generated in a VELOCIMMUNE® mouse. In one embodiment, VelocImmune® (VI) mice were immunized with DNA encoding the GP of the complete Ebola virus [Ebola Zairevirus 2014 (GenBank: KJ660346.2)]. Antibodies were generated after an accelerated regimen comprising 2 immunizations. Petition 870260058248, dated 06 / 15 / 2026, pp. 96 / 143 / 106, separated by 2 weeks. The antibody immune response was monitored by a specific immunoassay for Ebola virus GP. For example, sera were measured for titers of specific antibodies to purified complete EBOV GP, GP subunit proteins (GP1 and GP2), and virus-like particles (VLPs) expressing EBOV GP. Antibody-producing clones were isolated using both B-cell sorting technology (BST) and hybridoma methods. For example, when a desired immune response was achieved, splenocytes were collected and fused with mouse myeloma cells to preserve their viability and form hybridoma cell lines. The hybridoma cell lines were tested and screened to identify cell lines that produce specific antibodies to Ebola virus GP.Using this technique and the various immunogens described above, several chimeric antibodies were obtained (i.e., antibodies possessing human variable domains and mouse constant domains); the exemplary antibodies generated in this way were designated as H1M17354N, H2aM17356N, H1M17357N, H2aM17358N, H2aM17359N, and H2aM17360N.

[00204] Ebola antiviral antibodies were also isolated directly from non-fusion antigen-positive mouse B cells with myeloma cells, as described in U.S. Patent 7582298. Using this method, several completely human anti-GP Ebola virus antibodies were obtained (i.e., antibodies possessing both human variable domains and human constant domains were obtained); the exemplary antibodies generated in this way were designated H1H17134P, H1H17139P, H1H17142P, H1H17151P, H1H17161P, H1H17162P, H1H17193P, H1H17196P, H1H17199P, H1H17203P, H1H17214P, H1H17219P, H1H17223P, and H1H17228P.

[00205] The biological properties of the generated exemplary antibodies Petition 870260058248, dated 06 / 15 / 2026, pages 97 / 143 / 106, according to the methods in this example, are described in detail in the examples below. Example 2: Amino acid and nucleotide sequences of the variable region of the heavy chain and the light chain.

[00206] Table 1 establishes the amino acid sequence identifiers of the variable heavy and light chain regions and CDRs of selected Ebola antiviral antibodies of the invention. The corresponding nucleic acid sequence identifiers are defined in Table 2. Table 1: Amino acid sequence identifiers SEQ ID NOs: Antibody designation HCVR HCDR1 HCDR2 HCDR3 LCVR LCDR1 LCDR2 LCDR3 H1H17134P 2 4 6 8 10 12 14 16 H1H17139P 34 36 38 40 42 44 46 48 H1H17151P 50 52 54 56 58 60 62 64 H1H17161P 66 68 70 72 74 76 78 80 H1H17162 288 828 968 94 96 H1H17193P 98 100 102 104 106 108 110 112 H1H17196P 114 116 118 120 122 124 126 128 H1H17199P 131 301 133 433 140 142 144 H1H17203P 146 148 150 152 154 156 158 160 H1H17214P 162 164 166 168 170 172 174 176 H1H1 71 172 180 180 184 186 188 190 192 H1H17223P 194 196 198 200 202 204 206 208 H1H17228P 210 212 214 216 218 220 222 224H 17223H 228 230 232 234 236 238 240 H1H17356N 242 244 246 248 250 252 254 256 H1H17357N 258 260 262 264 264 266 270 272 H1H17358N2 274 276 278 280 282 284 286 288 H1H17359N 290 292 294 296 298 300 302 304 H1H17360N 306 383 283 283 286 288 H1M17354N 226 228 230 232 234 236 238 240 H2aM17356N 242 244 246 248 250 252 254 256 H1M1735 N 2735 2626 266 268 270 272H2aM17358N 274 276 278 280 282 284 286 288 H2aM17359N 290 292 294 296 298 300 302 304 Petition 870260058248, dated 06 / 15 / 2026, pages 98 / 143 / 106 I H2aM17360N 306 308 310 312 282 284 286 288 | Table 2: Nucleic acid sequence identifier_s SEQ ID NOs: Antibody designation HCVR HCDR1 HCDR2 HCDR3 LCVR LCDR1 LCDR2 LCDR3 H1H17134P 23 25 27 29 31 H1H17142P 33 35 37 39 41 43 45 47 H1H17151P 49 51 53 55 57 59 61 63 H1H17161P 65 67 69 71 77 79 79 H1H17162P 81 83 85 87 89 91 93 95 H1H17193P 97 99 101 103 105 107 109 111 H1H17196P H1H17199P 129 131 133 135 137 139 141 143 H1H17203P 145 147 149 151 153 155 157 159 H1H17214P 173 175 H1H17219P 177 179 181 183 185 187 189 191 H1H17223P 193 195 197 199 201 203 205 207 H1H17228 201 21 21 25 217 219 221 223 H1H17354N 225 227 229 231 233 235 237 239 H1H17356N 241 243 245 247 249 251 253 255 H1H17357 275 295 261 263 265 267 269 271 H1H17358N2 273 275 277 279 281 283 285 287 H1H17359N 289 291 293 295 297 297 209 30H136 H1 305 307 309 311 281 283 285 287 H1M17354N 225 227 229231 233 235 237 239 H2aM17356N 241 243 245 247 249 251 253 255 H1M17357N 257 259 261 263 265 267 269 271 H2aM17358N 273 275 277 279 281 283 285 287 H2aM17359N 289 291 293 295 297 299 301 303 H2aM17360N 305 307 309 311 281 283 285 287

[00207] Antibodies are typically referred to in the present invention according to the following nomenclature: Fc prefix (e.g., “H1H”, “H2M”, etc.) followed by a numerical identifier (e.g., “17139”, “17161”, etc., as shown in Table 1 or 2), followed by a suffix “P”, “P2”, “N”, N2 or “B”. The prefixes H1H and H2M on the antibody designations used in the present invention indicate the isotype of the Fc region. Petition 870260058248, dated 15 / 06 / 2026, p. 99 / 143 / 106 particular antibody. Thus, according to this nomenclature, an antibody may be referred to in the present invention as, for example, “H1H17359N”, “H2aM17359N”, etc. For example, an “H1M” antibody has a mouse IgG1 Fc, an “H2M” antibody has a mouse IgG2 Fc (isotype a or b) (all variable regions are fully human, as denoted by the first 'H' in the antibody designation). As will be noted by a person normally skilled in the art, an antibody with a specific Fc isotype can be converted to an antibody with a different Fc isotype (for example, an antibody with a mouse IgG1 Fc can be converted to an antibody with a human IgG4, etc.).However, in any case, the variable domains (including the CDRs) – which are indicated by the numerical identifiers shown in Table 1 or 2 – will remain the same, and the antigen-binding properties are expected to be identical or substantially similar regardless of the nature of the Fc domain. Example 3: Antibody binding to Ebola virus GP, as determined by surface plasmon resonance. A. pH-dependent dissociation rate constant at 37 °C

[00208] The dissociation rate constants (Kd) and dissociation half-lives (1½) for the binding of Ebola virus GP to Ebola virus anti-GP monoclonal antibodies at 37 °C were determined using a real-time surface plasmon resonance biosensor assay on a Biacore T200 instrument. The surface of the Biacore CM4 sensor was derivatized by amine coupling with either a monoclonal mouse anti-human Fc antibody (GE, # BR-1008-39) or a monoclonal goat anti-mouse Fc antibody (GE, # BR-1008-38) to capture Ebola virus anti-GP mAbs. All binding studies in Example 3A were performed in a buffer composed of 0.01M Na2HPO4 / NaH2PO4, 0.15M NaCl, and 0.05% v / v P20 surfactant. Petition 870260058248, dated 06 / 15 / 2026, pp. 100 / 143 88 / 106 (PBS-P analytical buffer) at pHs 7.4, 6.0, and 5.0. The search for low pH was performed to assess whether antibodies maintain binding at low pH. This would mimic the conditions the virus will encounter during membrane fusion after endosome acidification. Different concentrations of Ebola virus GP with a C-terminal polyhistidine marker (EbolaGP.his; Sino Biologicals, Catalog No. 40442-V08B1) prepared in PBS-P analytical buffer (ranging from 90 nM to 11.1 nM, 3-fold dilutions) were injected onto the captured surface of Ebola virus GP mAb at a flow rate of 25 pL / minute. The association of Ebola virus GP with captured monoclonal antibody was monitored for 5 minutes, and the dissociation of Ebola virus GP in PBS-P analysis buffer was monitored for 6 minutes. All dissociation rate constant experiments were performed at 37 °C.The kinetic dissociation rate constants (kd) were determined by fitting the sensograms in real time to a 1:1 linkage model using the Scrubber 2.0c curve fitting software. The dissociative half-lives (1½) were calculated from the kinetic rate constants as: ln(2) PA (min) = 6ü«kd

[00209] The dissociation rate parameters for the binding of Ebola virus GP to Ebola virus GP mAbs at 37 °C are shown in Table 3. Table 3: pH dependence of dissociative half-lives at 37 °C Ratio of t / 2 mAb captured pH 7.4 / pH 6.0 pH 7.4 / pH 5.0 H1H1238N(-) control NB NB H1H17162P 0.3 0.3 H1H17177P 0.2 0.2 H1H17193P 1.1 0.8 H1H17196P 1.0 1.0 H1H17150P 0.2 0.2 H1H17151P 0.03 0.01 Petition 870260058248, dated 06 / 15 / 2026, pages 101 / 143 / 106 H1H17160P 0.2 0.4 H1H17161P 0.2 0.2 H1H17214P 1.0 1.0 H1H17219P 1.0 1.0 H1H17223P 0.4 0.4 H1H17228P 0.6 0.14 H17214P 0.15 0.5 H1H17141P 0.3 0.3 H1H17139P 0.2 0.2 H1H17134P 0.6 0.6 H1H17211P 6.7 3.0 H1H17210P 0.2 0.2 H1H17203P 0.19 H1H17139P 0.19 0.4 0.1 H1M17348N 0.3 0.4 H1M17349N 2.7 6.9 H1M17350N 0.1 1.1 H1M17351N NB NB H1M17352N 1.1 1.1 H1M17353N 1.3M17349N 1.3 1.3 H1M17357N 0.8 0.5 H2aM17355N 0.5 0.6 H2aM17356N 0.9 0.9 H2aM17358N 0.7 0.4 H2aM17359N 0.8 0.7a H2aM17359N 0.8 0.7a H2aM17355N H2aM17361N 0.2 0.5 NB - undetectable binding under tested assay conditions B. Binding affinity and kinetics at 25 °C and 37 °C

[00210] Equilibrium dissociation constants (Kd values) for Ebola virus GP binding to Ebola virus anti-GP mAbs were determined using a real-time surface plasmon resonance biosensor, using a Biacore 4000 instrument. The surface of the Biacore CM4 sensor was derivatized by amine coupling with either a monoclonal mouse anti-human Fc antibody (GE, # BR-1008-39) or a monoclonal goat anti-mouse Fc antibody (GE, # BR-1008-38). Petition 870260058248, dated 06 / 15 / 2026, pp. 102 / 143 / 106 to capture anti-GP mAbs from the Ebola virus. All Biacore binding studies in example 3B were performed in a buffer composed of 0.0 IM HEPES, pH 7.4, 0.15M NaCl, 3 mM EDTA, 0.05% v / v P20 surfactant (HBS-ET analytical buffer). Different concentrations of Ebola virus GP with a C-terminal polyhistidine marker (Sino Biologicals, Catalog No. 40442-V08B1) prepared in HBS-ET analytical buffer (ranging from 90 nM to 3.3 nM, 3-fold dilutions) were injected onto the captured surface of the anti-Ebola virus GP mAb at a flow rate of 30 pL / minute. The binding of Ebola virus GP to the captured monoclonal antibody was monitored for 5 minutes, and the dissociation of Ebola virus GP in the HBS-ET analytical buffer was monitored for 10 minutes. All kinetic binding experiments were performed at 25 °C and 37 °C.The kinetic association rate (ka) and dissociation rate (Kd) constants were determined by fitting the sensograms in real time to a 1:1 linkage model using the Scrubber 2.0c curve fitting software. The equilibrium dissociation linkage constants (Kd) and dissociative half-lives (t%) were calculated from the kinetic rate constants, as follows: kd ln(2) Kd (M) = ka, and t'Z> (min) = 6ü*kd

[00211] The kinetic binding parameters for Ebola virus GP binding to Ebola virus GP mAbs at 25 °C and 37 °C are shown in Tables 4A and 4B. Table 4A: Binding kinetics at 25 °C mAb ka (1 / Ms) KD (1 / s) Kd(M) 11 / 2 (min) H1H17162P 2.18E+04 < 1E-5 4.60E-10 > 1155 H1H17177P 3.18E+03 l,12E-05 3.5-0393.30 H1H17193P 4.58E+03 l.08E-04 2.36E-08 106.6 H1H17196P 2.56E+04 < 1E-5 3.91E-10 > 1155 H1H17150P 2.16E+04E-02.05E-04 213.1 H1H17151P l,26E+04 9.44E-05 7.49E-09 122.3 H1H17160P 6.85E+04 3.76E-03 5.48E-08 3.1 Petition 870260058248, of 15 / 06 / 2026, p. 103 / 143 / 106 H1H17161P 5,29E+04 < 1E-5 1,89E-10 > 1155 H1H17214P 3,76E+04 < 1E-5 2,66E-10 > 1155 H1H17219P 3,11E+04 2,90E-05 9,34E-10 398,3 H1H17223P 3,00E+04 6,08E-05 2,03E-09 190,0 H1H17228P 4,49E+04 1,69E-03 3,76E-08 6,9 H1H17142P 2,00E+04 2,81E-05 1,41E-09 410,7 H1H17141P 1,98E+04 9,69E-05 4,90E-09 119,2 H1H17139P 2,29E+04 1,63E-04 7,13E-09 70,8 H1H17134P 7,65E+04 9,41E-04 1,23E-08 12,3 H1H17211P 3,33E+04 2,14E-04 6,43E-09 54,0 H1H17210P 1,09E+02 2,06E-04 1,89E-06 56,0 H1H17203P 2,78E+04 1,68E-04 6,04E-09 68,7 H1H17199P 1,25E+04 2,36E-04 1,89E-08 49,0 H1M17348N IC IC IC IC H1M17349N 7,03E+04 8,69E-04 1,24E-08 13,3 H1M17350N IC IC IC IC H1M17351N NB NB NB NB H1M17352N IC IC IC IC H1M17353N IC IC IC IC H1M17354N 4,94E+04 3,16E-03 6,39E-08 3,7 H1M17357N IC IC IC IC H2aM17355N 1,44E+04 < 1E-5 6,96E-10 > 1155 H2aM17356N 2,18E+04 9,57E-05 4,40E-09 120.7 H2aM17358N 3.22E+02 2.01E-04 6.23E-07 57.5 H2aM17359N 3.82E+03 1.95E-04 5.09E-08 59.4 H2aM17360N 2.30E+04 1.06E-05 4.63E-10 1086.5 H2aM17361N 1.22E+02 1.25E-04 1.02E-06 92.5. NB - connection not detectable under the tested conditions. IC - connection sensogram inconclusive for fit. Table 4B. Binding kinetics at 37 °C. mAb ka (1 / Ms) Kd (1 / s) Kd(M) t1 / 2 (min) H1H17162P 3.47E+04 < 1E-5 2.88E-10 > 1155 H1H17177P 1.68E+04 1.62E-04 9.62E-09 71.3 H1H17193P 1.58E+04 5.03E-04 3.18E-08 22.9 H1H17196P 3.16E+04 < 1E-5 3.17E-10 > 1155 H1H17150P 3.18E+04 3.94E-05 1.24E-09 292.8 H1H17151P 2.26E+04 3.83E-04 1.70E-08 30.2 H1H17160P 5.72E+04 5.63E-03 9.85E-08 2.1 H1H17161P 4.39E+04 < 1E-5 2.28E-10 > 1155 Petition 870260058248, dated 06 / 15 / 2026, pages 104 / 143 / 106 H1H17214P 3,67E+04 1,54E-04 4,20E-09 74,9 H1H17219P 4,41E+04 < 1E-5 2,27E-10 > 1155 H1H17223P 3,51E+04 2,42E-04 6,89E-09 47,7 H1H17228P 7,32E+04 3,83E-03 5,23E-08 3 H1H17142P 2,60E+04 1,74E-04 6,68E-09 66,6 H1H17141P 2,65E+04 2,92E-04 1,10E-08 39,6 H1H17139P 2,48E+04 5,12E-04 2,06E-08 22,5 H1H17134P 6,99E+04 4,69E-04 6,70E-09 24,6 H1H17211P 1,90E+04 7,31E-04 3,84E-08 15.8 H1H17210P 6.19E+02 6.12E-04 9.89E-07 18.9 H1H17203P 3.85E+04 1.19E-03 3.09E-08 9.7 H1H17199P 3.04E+04 1.28E-03 4.22E-08 9 H1M17348N IC IC IC IC H1M17349N 1.77E+04 1.93E-03 1.09E-07 6 ​​H1M17350N 4.84E+02 1.00E-03 2.07E-06 11.5 H1M17351N NB NB NB NB H1M17352N 4.09E+04 1.55E-03 3.80E-08 7.4 H1M17353N 2.33E+02 5.38E-04 2.31E-06 21.5 H1M17354N 5.08E+04 5.73E-03 1.13E-07 2 H1M17357N 2.35E+04 1.84E-03 7.81E-08 6.3 H2aM17355N 1.99E+04 2.06E-04 1.03E-08 56.2 H2aM17356N 7.26E+03 2.50E-04 3.44E-08 46.2 H2aM17358N 1.07E+04 5.67E-04 5.28E-08 20.4 H2aM17359N 1.54E+04 3.52E-04 2.29E-08 32.8 H2aM17360N 2.43E+04 3.37E-04 1.39E-08 34.3 H2aM17361N 1.83E+04 4.15E-04 2.27E-08 27.8. NB - connection not detectable under the tested conditions. IC - connection sensogram inconclusive for fit. Results

[00212] As shown in Tables 4A and 4B above, antibodies bound to Ebola virus GP with KD values ​​ranging from 934 pM to 1890 nM at 25°C, and from 227 pM to 2310 nM at 37°C. At pH 7.4, antibodies showed dissociative half-life (1½) values ​​ranging from 3.0 minutes to more than 1155 minutes at 25°C, and from 2.0 minutes to more than 1155 minutes at 37°C. No loss of binding was observed at low pH. Several antibodies showed increased dissociative half-life (1½) values ​​at low pH compared to pH 7.4. Antibodies with increases of 3 Petition 870260058248, dated 06 / 15 / 2026, pp. 105 / 143 / 106 times or more in the dissociative half-life values ​​(1½) at pH 5 and / or pH 6 include H1H17162P, H1H17177P, H1H17150P, H1H17151P, H1H17160P, H1H17161P, H1H17141P, H1H17139P, H1H17210P, H1H17203P, H1H17199P, H1M17348N, H1M17350N, H2aM17360N and H2aM17361N. Example 4: Generation of Ebola virus pseudoparticles and neutralization studies

[00213] Ebola virus pseudoparticles (also called virus-like particles or VLPs) were generated by co-transfecting 293T cells with a mixture of plasmid constructs expressing Ebola virus GP, HIV gag-pol, and an HIV proviral vector encoding firefly luciferase. Supernatants containing Ebola virus pseudoparticles were collected 48 hours after transfection, clarified using centrifugation, aliquoted, and frozen at -80 °C. Control pseudoparticles were generated by replacing the Ebola virus GP-expressing plasmid with a plasmid encoding vesicular stomatitis virus glycoprotein (VSVg). Neutralization assay based on Ebola pseudoparticle

[00214] The pseudoparticles generated, as described above, were tested in neutralization assays. Specifically, antibody dilutions were incubated with Ebola virus pseudoparticles for 1 h at room temperature. Huh7 cells were released using 0.02 M EDTA, washed, and incubated with the antibody / pseudoparticle mixtures for 72 h. Infection efficiency was quantified by luciferase detection using the BrightGlo® luciferase assay (Promega, San Luis Obispo, CA, USA) and read on a Victor® X3 plate reader (Perkin Elmer, Waltham, MA, USA) to produce light. Table 5: Neutralization of the Zaire-type VLP 2014 ID AB ID of the corresponding hybridoma Ab Neutralizer of the Zaire type VLP 2014 IC50(M) H1H17134P - - Petition 870260058248, dated 06 / 15 / 2026, page 106 / 143 / 106 H1H17139P - - H1H17142P + 1.59E-09 H1H17151P + 1.51E-09 H1H17161P + 2.55E-10 H1H17162P + 2.86E-10 H1H17193P - - H1H17196P + 1.68E-09 H1H17199P - - H1H17203P + 8.68E-10 H1H17214P + 8.99E-10 H1H17219P + 6.95E-10 H1H17223P + 1.58E-09 H1H17228P + 3.26E-09 H1M17354N H1M17354N - - H2aM17356N H2aM17356N + 4.77E-09 H1M17357N H1M17357N - - H2aM17358N H2aM17358N + 4.68E-09 H2aM17359N H2aM17359N + 3.36E-09 H2aM17360N H2aM17360N + 3.75E-09

[00215] The data shown above in Table 5 show that 14 of the 20 Ebola antiviral antibodies of the present invention, using the experimental design described in this document, potentially neutralize infectivity with an IC50 ranging from about 10-11M to about 10-9M. Example 5: Antibody-dependent cell-mediated cytotoxicity (ADCC) by Ebola antiviral antibodies

[00216] Antibody-dependent cell-mediated cytotoxicity (ADCC) was tested by the ability of antibodies to signal through a CD16-based reporter system (Promega ADCC Reporter Bioassay Essential Kit, San Luis Obispo, CA, USA). 293 cells expressing Ebola virus GP were seeded. One day later, diluted antibodies produced on fuc- cell lines (see U.S. Patent No. 8409838) and effector cells (effector-to-target ratio of 1.5:1) were added and incubated overnight. Reporter activity was measured by the BrightGlo® luciferase assay (Promega, San Luis Obispo, USA). Petition 870260058248, dated 06 / 15 / 2026, page 107 / 143 / 106 CA, USA) and read on a Victor® X3 plate reader (Perkin Elmer, Waltham, MA, USA) in relation to light output. Table 6: ADCC Results ADCC ID AB Reporter Bioassay Clone ID ADCC Activity H1H17134P + H1H17139P + H1H17142P + H1H17151P + H1H17161P - H1H17162P - H1H17193P + H1H17196P + H1H17199P + H1H17203P + H1H17214P + H1H17219P - H1H17223P + H1H17228P + H1M17354N HCAF05C08-22 + H2aM17356N HCAF08C07-09 + H1M17357N HCAF09D11-13 + H2aM17358N HCAF12C05-14+ H2aM17359N HCAF12C06-26 + H2aM17360N HCAF12G09-07 +

[00217] The ability of antibodies to mediate ADCC was calculated based on activity compared to an isotype control (negative). Any value greater than 5 times above the negative control was considered positive. The data above in Table 6 show that 17 of the 20 Ebola antiviral antibodies mediated ADCC. Example 6: Cross-competition in Octet

[00218] The binding competition between Ebola virus GP monoclonal antibodies that were previously determined to bind to Ebola virus GP was determined using a real-time label-free biolayer interferometry (BLI) assay on an Octet HTX biosensor. Petition 870260058248, dated 06 / 15 / 2026, pp. 108 / 143 / 106 (ForteBio Corp., a division of Pall Life Sciences). The binding of the relevant controls to soluble GP (sGP), GP1 or GP2, was measured in the same assay format, and its response was subtracted from the GP reagent of the Ebola virus of interest for each mAb tested. The entire experiment was performed at 25 °C in buffer comprising 0.01M HEPES pH 7.4, 0.15M NaCl, 3 mM EDTA, 0.05% v / v surfactant P20, 1.0 mg / mL BSA (HBS-ET octet buffer), with the plate under agitation at a speed of 1000 rpm. To assess whether two antibodies are capable of competing with each other for binding to their respective epitopes in Ebola virus GP expressed with a C-terminal polyhistidine marker (Ebola virus GP.h, Sino Biologicals Inc., see also AHX24649.1 in GenBank and SEQ ID NO: 314), approximately ~1.0 nm of Ebola virus GP was first captured on octet biosensors coated with anti-penta-His antibody (Fortebio Inc., No.(18-5079) by submerging the biosensors for 3 minutes in wells containing a 20 pg / mL solution of Ebola virus GP. The antigen-captured biosensors were then saturated with the first anti-Ebola virus GP monoclonal antibody (hereinafter referred to as mAb-1) by immersion in wells containing a 50 pg / mL solution of mAb-1 for 5 minutes. The biosensors were then subsequently submerged in wells containing a 50 pg / mL solution of a second anti-Ebola virus GP monoclonal antibody (hereinafter referred to as mAb-2) for 3 minutes. All biosensors were washed in HBS-ET octet buffer between each step of the experiment. The real-time binding response was monitored throughout the experiment, and the binding response at the end of each step was recorded.The binding response of mAb-2 to Ebola virus GP pre-complexed with mAb-1 was compared, and the competitive / non-competitive behavior of different anti-Ebola virus GP monoclonal antibodies was determined using a 50% inhibition threshold. Table 7 explicitly defines the relationships of the competing antibodies in both directions. Petition 870260058248, dated 06 / 15 / 2026, page 109 / 143 / 106 regardless of the order of connection.

[00219] As shown in Table 7, the left column shows that mAb1 antibodies are captured using AHC octet biosensors and the right column shows the antibodies (mAb2) that cross-compete with the mAb1 antibody. Table 7: Cross-competition of anti-GP antibodies to the Ebola virus for Ebola virus GP binding. First mAb (mAb-1) captured using AHC octet biosensors mAb-2 antibodies that were shown to compete with mAb-1 H1H17160P H1H17160P, H1M17354N, H1M17357N, H1H17228P, H1M1735 H1H17160P, H1M17354N, H1M17357N, H1H17228P, H1H17203P H1M17357N H1H17160P, H1M17354N, H1M17357N, H1H17228P, H1H17357N, H1H17228P, H1H1722H2 H1H17160P, H1M17354N, H1M17357N, H1H17228P, H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N H1H17203PH17160, H1M17354N, H1M17357N, H1H17228P, H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H1719P, H1H17214P, H1H17199, H1H1728P, H1H17151P H2aM17360N, H1M17352N H1H17151P H1H17228P, H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H1721, H1H17159N, H1H1721, H1H17151, H1H17191 H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17353N, H1H17223P, H1PH19, H1719H H1H17142P H1H17228P, H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358, H1H17228P, H2aM17203N, H1H17142P H1M17352N, H2aM17356N, H2aM17361N, H2aM17355N, H1M17353N, H1H17223P,H1H17196P, H1H17193P H1H17177P H1H17228P, H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17348N, H1M17353N, H1H17141P, H1H17223P, H1H17196P, H1H17139P, H1H17193P, H1M17350N H2aM17359N H1H17228P, H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N H2aM17356N, H2aM17361N, H2aM17355N, 1H17211P, H1M17348N, H1M17353N, H1H17141P, H1H17223P H1H17139P, H1H17193P, H1M17350N H1H17214P H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P H1H17199P, H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N H2aM17361N, H2aM17355N, H1H17211P, H1M17348N, H1M17353N H1H17141P, H1H17223P, H1H17139P H1H17193P, H1M17350N, Petition 870260058248, dated 06 / 15 / 2026, pages 110 / 143 / 106 H1H17199P H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P H1H17199P, H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17348N, H1M17353N, H1H17141P, H1H17139P, H1H17193P, H1M17350N H2aM17358N H1H17203P, H1H17151P H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N H2aM17361N, H2aM17355N, H1H17211P, H1M17348N, H1M17353N H1H17139P, H1H17193P, H1M17350N H2aM17360N H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17348N H1M17353N, H1H17139P, H1H17193P, H1M17350N H1M17352N H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17348N, H1M17353N, H1H17139P, H1H17193P, H1M17350N H2aM17356N H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P H1H17199P, H2aM17358NH2aM17360N, H1M17352N, H2aM17356N H2aM17361N, H2aM17355N, H1H17211P, H1M17348N H1M17353N, H1H17139P, H1H17193P, H1M17350N H2aM17361N H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N H2aM17355N, H1H17211P, H1M17348N, H1M17353N, H1H17139P, H1H17193P, H1M17350N H2aM17355N H1H17203P, H1H17151P, H1H17142P, H1H17177P, H2aM17359N H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17348N H1M17353N, H1H17193P, H1M17350N H1H17211P H1H17151P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17348N, H1M17353N H1H17193P, H1M17350N H1M17348N H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N H2aM17355N, H1H17211P, H1M17348N, H1M17353N, H1H17193P H1M17350N H1M17353N H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P H1H17199P, H2aM17358NH2aM17360N, H1M17352N H2aM17356N, H2aM17361N, H2aM17355N, H1H17211P, H1M17348N, H1M17353N, H1H17141P, H1H17223P, H1H17196P, H1H17139P, H1H17193P H1H17141P H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H1M17353N H1H17141P, H1H17223P, H1H17196P, H1H17139P H1H17223P H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1M17353N H1H17141P, H1H17223P, H1H17196P, H1H17139P H1H17196P H1H17151P, H1H17142P, H1H17177P, H1M17353N, H1H17141P, H1H17223P, H1H17196P, H1H17139P H1H17139P H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N, H1M17353N H1H17141P, H1H17223P, H1H17196P, H1H17139P H1H17193P H1H17151P, H1H17142P, H1H17177P, H2aM17359N, H1H17214P, H1H17199P H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N H2aM17355N, H1H17211P, H1M17348N, H1M17353N, H1H17193P, H1M17350N, Petition 870260058248, dated 06 / 15 / 2026, pages 111 / 143 / 106 H1M17350N H1H17177P, H2aM17359N, H1H17214P, H1H17199P, H2aM17358N, H2aM17360N, H1M17352N, H2aM17356N, H2aM17361N, H2aM17355N H1H17211P, H1M17348N, H1H17193P, H1M17350N H1H17219P H1H17219P, H1H17150P, H1H17161P H1H17150P H1H17219P, H1H17150P, H1H17161P H1H17161P H1H17219P, H1H17150P, H1H17161P H1M17349N H1M17349N H1H17134P H1H17134P H1H17162P H1H17162P H1H17210P H1H17210P Example 7: Sequential binding of H1H17203P, H1H17139P, and H1H17161P to the Ebola virus glycoprotein.

[00220] Based on the information obtained from the cross-competition experiments, a sequential binding study was conducted to determine if three individual candidate antibodies are capable of simultaneously binding to the soluble glycoprotein of the Ebola virus (GP), thereby confirming that the binding sites on the GP of the Ebola virus are independent for each monoclonal antibody. If so, this information would support the use of these antibodies in a therapeutic cocktail.

[00221] In this sense, sequential binding experiments for three Ebola virus anti-GP monoclonal antibodies, H1H17203P, H1H17139P, and H1H17161P, were tested for independent and non-competitive binding with Ebola virus GP. This experiment was performed using a real-time label-free biolayer interferometry (BLI) assay on an Octet RED biosensor (ForteBio Corp., a division of Pall Life Sciences). The entire experiment was conducted at 25 °C in buffer comprising 0.01M HEPES pH 7.4, 0.15M NaCl, 3 mM EDTA, 0.05% v / v P20 surfactant, and 1.0 mg / mL BSA (HBSET octet buffer), with the plate under agitation at a speed of 1000 rpm. To assess whether the three antibodies can simultaneously bind to the GP of captured Ebola virus antigen expressed with a C-terminal polyhistidine marker (ebola virus GP.his, Sino Biologicals), approximately ~0.6 nm from the GP of Petition 870260058248, dated 06 / 15 / 2026, pp. 112 / 143100 / 106 ebola.h virus was first captured on Octet biosensors coated with anti-penta-His antibody (Fortebio Inc, No. 18-5079) by submerging the biosensors for 3 minutes in wells containing a GP solution of ebola.h virus. The antigen-captured biosensors were then saturated with the first anti-GP monoclonal antibody of ebola virus (later referred to as H1H17161P) by immersion in wells containing a 50 pg / mL solution of REGN H1H17161P for 5 minutes. The biosensors were then subsequently submerged in wells containing a 50 pg / mL solution of a second anti-GP monoclonal antibody of ebola virus (later referred to as H1H17139P) for 5 minutes. Finally, 50 ug / mL of the third antibody (later referred to as H1H17161P) was injected over 5 minutes to achieve saturation. The real-time binding response was monitored throughout the experiment, and the binding response at the end of each step was recorded.Results.

[00222] The three candidate monoclonal antibodies tested were able to bind simultaneously to the GP of the Ebola virus, indicating that each antibody did not interfere with the binding site on the GP of the Ebola virus of the other antibodies tested, suggesting that each of them bound or interacted with different epitopes. This supports a role for the use of these three antibodies in a therapeutic antibody cocktail. Example 8: Binding of anti-Ebola antibodies to different strains of Ebola virus-like particles (VLPs)

[00223] A study was conducted to determine whether anti-GP antibodies to the Ebola virus could react with virus-like particles (VLPs) containing GPs from other Ebola virus strains. This study included VLPs containing GPs of the Bundibugyo type NC_014373, Cote d'Ivoire FJ217162, Sudan NC_006432, Zaire.1995, Zaire.2014 AY354458 and a negative control, the VSV glycoprotein. Petition 870260058248, dated 06 / 15 / 2026, pages 113 / 143 101 / 106

[00224] (VSVg). The study was conducted using “MesoScale Discovery” (MSD), a technology that allows the binding / attachment of Ebola strain VLPs (which express the Ebola viral surface glycoproteins) to a carbon surface, followed by an ELISA-type binding assay. The objective was to identify the mAb binding profiles in relation to the various Ebola strains.

[00225] The assay was performed in 96-well polypropylene microwell plates by first preparing a 1:10 dilution of the supernatants from several VLPs / well (as observed in the following table) and adding the dilutions to PBS (50 gL / well) and incubating at 4 °C overnight.

[00226] The liquid from the wells was discarded, followed by blocking with 150 gL / well in PBS + 2% BSA and incubation for one hour at room temperature. The contents of each well were then discarded, and the wells were washed with PBS using an AquaMax2000 plate washer designated for MSD. Fifty microliters of the primary antibody were diluted in PBS + 1% BSA and incubated at room temperature with agitation at an intermediate speed (5). The well contents were then discarded, and the plates were washed with PBS. Fifty microliters of sulfo-TAG detection reagent (human or mouse Fc) at a concentration of 1 gg / mL of PBS + 0.5% BSA were added to each well and incubated at room temperature for one hour with agitation at an intermediate speed. The well contents were discarded, and the plates were washed with PBS + 0.5% BSA.150 gL of 1X surfactant-free reading buffer was added to each well, and the plates were scanned using a SECTORImager6000 barcode scanner.

[00227] The results, shown in Table 8 below, demonstrate that all tested anti-GP antibodies to the Ebola virus bind to VLPs containing Zaire 2014 and Zaire 1995 type GPs. Some of the antibodies Petition 870260058248, dated 06 / 15 / 2026, pp. 114 / 143 102 / 106 tested bind to VLPs containing GPs from other Ebola virus strains, in addition to binding to the two Zaire-type strains observed in Table 8. In particular, in addition to binding to VLPs containing the Zaire 2014 and Zaire 1995 GPs, the Ebola antiviral antibodies designated H1H17161P and H1H17162P bind to VLPs containing the GPs of the Sudan and Bundibugyo strains, while the Ebola antiviral antibodies designated H2aM17356N and H1H17142P bind to the Bundibugyo and Cote d'Ivoire strains. Table 8: Cross-reactivity of Ebola antiviral antibodies with GPs from various Ebola virus strains_______________________________________________ Ebola antiviral antibody binding to VLPs containing GPs from various Ebola virus strains AbPID Sudan Bundibugyo Cote d'Ivoire Zaire 2014 Zaire 1995 H1H17161P + + _ + + H1H17139P _ _ _ + + H1H17203P _ _ _ + + H1H17219P _ _ _ + + H1H17162P + + _ + + H1H17199P _ _ _ + + H1H17193P _ _ _ + + H1M17354N _ _ _ + + H1M17357N _ _ _ + + H1H17134P _ _ _ + + H1H17360N _ _ _ + + H1H17358N2 _ _ _ + + H2aM17356N _ + + + + H1H17223P _ _ _ + + H1H17196P _ _ _ + + H1H17151P _ _ _ + + H1H17142P _ + + + + H1H17214P _ _ _ + + H1H17228P _ _ _ + + H2aM17359N _ _ _ + + Example 9. In vitro neutralization of live / infectious Ebola virus (EBOV)

[00228] Antibodies designated as H1H17203P, H1H17139P and H1H17161P were analyzed for their ability to neutralize the Petition 870260058248, dated 06 / 15 / 2026, pp. 115 / 143 103 / 106 EBOV in Vero cells. Vero cells were plated in 384-well FBS plates with 10% DMEM and allowed to grow to approximately 75% confluence at 37°C. H1H17203P, H1H17139P, and H1H17161P were diluted as directed. EBOV strains (Mayinga, Kikwit, Makona, and Mayinga adapted for guinea pig) were thawed and diluted appropriately to an MOI between 0.01 and 0.1. A commercially available anti-EBOV antibody designated KZ52 was used as a positive control. (See Maruyama, T. et al., J Virol 73, 6024-6030 (1999). The antibodies were incubated with the virus for 1 hour at 37°C. The antibody / virus mixture was then added to pre-plated cells and the plates were incubated at 37°C for 24 hours. After the incubation period, the plates were removed from the incubator and inactivated by immersion in 10% neutral buffered formalin, placed in a sealed bag and stored at 4°C overnight in BSL-4.)The plates were washed three times in 1X-PBS and the cells were permeabilized at room temperature (RT) with 25 pL of 0.1% Triton X-100 in 1X-PBS for 15–20 minutes. Triton X was discarded and the plates were blocked with 3.5% BSA in 1X-PBS for 1 hour at room temperature. The plates were treated overnight at 4 °C with GP anti-EBOV mouse primary antibody 4F3 (catalog number 0201-020) diluted 1:1500 in 1X-PBS. The plates were washed in 1X-PBS for 10–15 minutes and again twice more. The cells were incubated for 1 hour with Alexa-fluor-488-conjugated anti-mouse secondary antibody. The secondary antibody was discarded, and the plates were washed in 1X-PBS for 10–15 minutes and again twice more. The plates were incubated with 25 pI / well of Hoechst (1:50,000 in 1X-PBS) for 30 minutes at room temperature.The plates were photographed by fluorescence microscopy using the blue and green fluorescence channels. Petition 870260058248, dated 06 / 15 / 2026, pages 116 / 143 104 / 106 Results

[00229] The results, shown in Figure 1, demonstrated that the antibody H1H17161P neutralized the live virus and was more potent than the positive control antibody KZ52, but the antibodies designated as H1H17203P and H1H17139P did not act as neutralizers. Example 10: Binding of anti-Ebola antibodies to soluble GP (sGP)

[00230] The fourth gene in the EBOV genome encodes two unique proteins, a non-structural dimeric secreted glycoprotein, named sGP, and a virion-linked trimeric envelope glycoprotein (GP). These two GPs share the first 295 amino acids but have unique C-terminuses. To determine if Regeneron mAbs bind to sGP, a recombinant sGP.mmh protein was produced in-house (SEQ ID NO: 317). The Octet HTX interferometry-based biosensor was used to determine if the monoclonal antibodies H1H17203P, H1H17139P, and H1H17161P can bind to the Ebola sGP.mmh protein. The assay format involved capturing H1H17203P, H1H17139P, and H1H17161P at the tips of the anti-hFc sensor, followed by immersion in 300 nM solutions of GP.10xhis from Ebola (SEQ ID NO: 318), sGP.mmh (SEQ ID n: 317), or hCNTFR (ciliary neurotrophic factor receptor.mmh, which is a negative control protein).Each mAb was captured at a level between 0.94 and 1.36 nm.

[00231] As shown in Figure 2, all mAbs showed specific binding to ebola GP.10xhis, and no binding to the negative control protein; while only H1H17139 demonstrated specific binding to ebola sGP.mmh. This finding suggests that the binding epitope of H1H17139 is likely located in a region common to the first 295 amino acids of sGP and GP; while the other mAbs possibly only recognize the C-terminal of ebola GP. Petition 870260058248, dated 06 / 15 / 2026, pp. 117 / 143 105 / 106 Example 11: Binding of additional anti-GP antibodies from EBOV to GP.h of ebola, soluble GP.mmh of ebola and hCNTFR.mmh

[00232] A more in-depth study was conducted to determine the binding characteristics of the additional anti-EBOV GP antibodies of the invention; in particular, the study was performed to determine the ability of these additional antibodies to bind to soluble GP and GP. This study was performed using a real-time label-free biolayer interferometry (BLI) assay on an Octet HTX biosensor (ForteBio Corp., a division of Pall Life Sciences). The entire experiment was performed at 25 °C in buffer comprising 0.01M HEPES pH 7.4, 0.15M NaCl, 3 mM EDTA, 0.05% v / v P20 surfactant, 1.0 mg / mL BSA (HBS-ET octet buffer), with the plate under agitation at a speed of 1000 rpm. To assess whether antibodies were able to bind to Ebola sGP or other Ebola GP reagents, approximately ~1.0 nm of anti-Ebola GP mAbs were captured on Octet biosensors coated with anti-human Fc antibody (Fortebio Inc, No.18-5064) by submerging the biosensors for 3 minutes in wells containing mAb solutions at 20 pg / mL. The biosensors captured by the mAb were tested for binding to selected protein reagents by immersion in wells containing 300 nM solutions of Ebola GP proteins or irrelevant controls for 5 minutes. All biosensors were washed in HBS-ET octet buffer between each step of the experiment. The real-time binding response was monitored throughout the experiment, and the binding response at the end of each step was recorded. Results

[00233] As shown in Table 9, any value below 0.10 nm was determined to be a non-binding antibody. Based on the results to date, all but one of the antibodies (H1H17360N) tested demonstrated complete GP binding to EBOV, and thirteen of the twenty Petition 870260058248, dated 06 / 15 / 2026, pages 118 / 143 106 / 106 antibodies tested showed binding to soluble GP (sGP). Table 9: Binding of anti-GP antibodies to GP.h of ebola, soluble GP.mmh of ebola and hCNTFR.mmh Antibody Number Binding to sGP Binding to 300 nM sGP of Ebola (F2) (nm) Binding to 300 nM GP.h of Ebola (nm) Binding to 300 nM hCNTFR.mmh (negative control) (nm) H1H17161P No 0.00 0.55 -0.01 H1H17139P Yes 0.19 0.55 0.01 H1H17203P No 0.02 0.61 0.01 H1H17219P No 0.01 0.68 0.01 H1H17162P No 0.03 0.49 0.02 H1H17199P Yes 0.33 0.38 0.00 H1H17193P Yes 0.26 0.33 0.02 H1M17354N Yes 0.18 0.71 0.02 H1M17357N No 0.10 0.56 0.01 H1H17134P No 0.01 0.70 -0.01 H1H17360N No 0.09 0.09 0.03 H1H17358N2 Yes 0.30 0.35 0.01 H1H17356N Yes 0.22 0.23 0.02 H1H17223P Yes 0.31 0.61 0.02 H1H17196P Yes 0.25 0.62 0.01 H1H17151P Yes 0.33 0.43 -0.02 H1H17142P Yes 0.28 0.34 0.01 H1H17214P Yes 0.38 0.52 0.01 H1H17228P Yes 0.35 0.58 0.00 H1H17359N Yes 0.39 0.51 0.00 Petition 870260058248, dated 06 / 15 / 2026, pp. 119 / 143

Claims

1 / 9 CLAIMS 1. Isolated recombinant antibody that binds specifically to Ebola virus (EBOV) and / or an Ebola virus glycoprotein (EBOV-GP), characterized in that the antibody comprises three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained in the heavy chain variable region / light chain variable region (HCVR / LCVR) amino acid sequence pair in SEQ ID Nos: 34 / 42.

2. Recombinant antibody isolated according to claim 1, characterized in that it is an immunoglobulin molecule comprising two heavy chains and two light chains linked by disulfide bonds, wherein each heavy chain comprises an HCVR comprising HCDR1, HCDR2 and HCDR3, and each light chain comprises an LCVR comprising LCDR1, LCDR2 and LCDR3.

3. Recombinant antibody isolated according to claim 1 or 2, characterized in that it comprises an amino acid sequence HCDR1 of SEQ ID NO: 36; an amino acid sequence HCDR2 of SEQ ID NO: 38; an amino acid sequence HCDR3 of SEQ ID NO: 40; an amino acid sequence LCDR1 of SEQ ID NO: 44; an amino acid sequence LCDR2 of SEQ ID NO: 46 and an amino acid sequence LCDR3 of SEQ ID NO:

48.

4. Recombinant antibody isolated according to any one of claims 1 to 3, characterized in that it comprises an HCVR having an amino acid sequence of SEQ ID NO:

34.

5. Recombinant antibody isolated according to any one of claims 1 to 3, characterized in that it comprises an LCVR having an amino acid sequence of SEQ ID NO:

42.

6. Recombinant antibody isolated according to any Petition 870260058248, dated 06 / 15 / 2026, p. 120 / 143 2 / 9 one of claims 1 to 3, characterized in that it comprises a pair of HCVR / LCVR amino acid sequences of SEQ ID NOS: 34 / 42.

7. Use of the antibody as defined in any one of claims 1 to 6, characterized in that it is for the manufacture of a medicament to treat or prevent infection with EBOV.

8. Use according to claim 7, characterized in that the medicament further comprises one or more additional therapeutic agents, which are selected from the group consisting of an antiviral drug, an anti-inflammatory drug (e.g., corticosteroids and non-steroidal anti-inflammatory drugs), a different antibody for EBOV, a vaccine for EBOV, TKM Ebola (small interfering RNAs that target viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (presensive morpholino oligomers of phosphorodiamidates that target the VP24 gene of EBOV) and interferons.

9. Use according to claim 8, characterized in that one or more additional therapeutic agents comprise one or more anti-EBOV antibodies.

10. Use according to claim 9, characterized in that one or more anti-EBOV antibodies comprise a pair of amino acid sequences of HCVR and LCVR selected from the group consisting of SEQ ID NO: 2 / 10, 18 / 26, 50 / 58, 66 / 74, 82 / 90, 98 / 106, 114 / 122, 130 / 138, 146 / 154, 162 / 170, 178 / 186, 194 / 202, 210 / 218, 226 / 234, 242 / 250, 258 / 266, 274 / 282, 290 / 298 and 306 / 282.

11. Use according to claim 10, characterized in that one or more anti-EBOV antibodies comprise a pair of amino acid sequences of HCVR and LCVR selected from the group consisting of SEQ ID NOs: 18 / 26, 66 / 74 and 146 / 154.

12. Use according to claim 10, characterized by Petition 870260058248, dated 06 / 15 / 2026, p. 121 / 143 3 / 9 fact that one or more anti-EBOV antibodies comprise an HCVR amino acid sequence of SEQ ID NO: 18 and an LCVR amino acid sequence of SEQ ID NO:

26.

13. Use according to claim 10, characterized in that one or more anti-EBOV antibodies comprise an HCVR amino acid sequence of SEQ ID NO: 66 and an LCVR amino acid sequence of SEQ ID NO:

74.

14. Use according to claim 10, characterized in that one or more anti-EBOV antibodies comprise an HCVR amino acid sequence of SEQ ID NO: 146 and an LCVR amino acid sequence of SEQ ID NO:

154.

15. Pharmaceutical composition, characterized in that it comprises one or more isolated recombinant antibodies that bind specifically to EBOV and / or EBOV-GP as defined in any of claims 1 to 6 and a pharmaceutically acceptable carrier or diluent.

16. Pharmaceutical composition according to claim 15, characterized in that it further comprises one or more isolated monoclonal antibodies that bind specifically to EBOV and / or EBOVGP, each comprising a pair of HCVR / LCVR amino acid sequences selected from the group consisting of SEQ ID NOs: 18 / 26, 66 / 74 and 146 / 154.

17. Pharmaceutical composition, characterized in that it comprises (a) a first anti-EBOV antibody as defined in any one of claims 1 to 6; (b) a second anti-EBOV antibody; and (c) a third anti-EBOV antibody; and (d) a pharmaceutically acceptable carrier or diluent.

18. Pharmaceutical composition according to claim 17, characterized in that the first antibody binds or interacts with a first epitope in EBOV, and the second and / or third antibody binds or interacts with a different epitope in EBOV.

19. Pharmaceutical composition according to claim 17 or 18, characterized in that each of the second and third anti-EBOV antibodies comprises a pair of HCVR / LCVR amino acid sequences selected from the group consisting of the sequences SEQ ID NOs: 2 / 10, 18 / 26, 50 / 58, 66 / 74, 82 / 90, 98 / 106, 114 / 122, 130 / 138, 146 / 154, 162 / 170, 178 / 186, 194 / 202, 210 / 218, 226 / 234, 242 / 250, 258 / 266, 274 / 282, 290 / 298 and 306 / 282.

20. Pharmaceutical composition according to claim 17 or 18, characterized in that the epitopes to which the first, second and / or third anti-EBOV antibody bind or interact are distinct and non-overlapping.

21. Pharmaceutical composition according to claim 17 or 18, characterized in that the first anti-EBOV antibody binds to or interacts with an epitope on an EBOV strain and the second and / or third anti-EBOV antibody binds to or interacts with a second and / or third epitope on the same strain or on a different EBOV strain.

22. Pharmaceutical composition according to claim 21, characterized in that the EBOV strains are selected from the group consisting of the strains Zaire.2014, Zaire.1995, Sudan, Bundibugyo and Cote d'Ivoire.

23. Pharmaceutical composition, characterized in that it comprises a first isolated monoclonal antibody that binds specifically to EBOV, wherein the first isolated monoclonal antibody comprises an amino acid sequence HCDR1 of SEQ ID NO: 36; an amino acid sequence HCDR2 of SEQ ID NO: 38; an amino acid sequence HCDR3 of SEQ ID NO: 40; an amino acid sequence LCDR1 of SEQ ID NO: 44; an amino acid sequence LCDR2 of SEQ ID NO: 46 and an amino acid sequence LCDR3 of SEQ ID NO: 48, and a pharmaceutically acceptable carrier or diluent.

24. Pharmaceutical composition according to claim 23, characterized in that it comprises a first isolated monoclonal antibody that binds specifically to EBOV, wherein the first isolated monoclonal antibody comprises an amino acid sequence HCDR1 of SEQ ID NO: 20; an amino acid sequence HCDR2 of SEQ ID NO: 22; an amino acid sequence HCDR3 of SEQ ID NO: 24; an amino acid sequence LCDR1 of SEQ ID NO: 28; an amino acid sequence LCDR2 of SEQ ID NO: 30 and an amino acid sequence LCDR3 of SEQ ID NO:

32.

25. Pharmaceutical composition according to claim 23, characterized in that it further comprises a second isolated monoclonal antibody that binds specifically to EBOV, wherein the second isolated monoclonal antibody comprises an HCDR1 amino acid sequence from SEQ ID NO: 68; an HCDR2 amino acid sequence from SEQ ID NO: 70; an HCDR3 amino acid sequence from SEQ ID NO: 72; an LCDR1 amino acid sequence from SEQ ID NO: 76; an LCDR2 amino acid sequence from SEQ ID NO: 78 and an LCDR3 amino acid sequence from SEQ ID NO:

80.

26. Pharmaceutical composition according to claim 23, characterized in that it further comprises a third isolated monoclonal antibody that specifically binds to EBOV, wherein the third isolated monoclonal antibody comprises an HCDR1 amino acid sequence from SEQ ID NO: 148; an HCDR2 amino acid sequence from SEQ ID NO: 150; an HCDR3 amino acid sequence from SEQ ID NO: 152; an LCDR1 amino acid sequence from SEQ ID NO: 156; an LCDR2 amino acid sequence from SEQ ID NO: 158 and an LCDR3 amino acid sequence from SEQ ID NO:

160.

27. Pharmaceutical composition according to any one of claims 23 to 26, characterized in that the first isolated monoclonal antibody is an immunoglobulin molecule comprising two heavy chains and two light chains linked by disulfide bonds, wherein each heavy chain comprises an HCVR comprising domains HCDR1, HCDR2 and HCDR3, and each light chain comprises an LCVR comprising domains LCDR1, LCDR2 and LCDR3.

28. Pharmaceutical composition according to any one of claims 24 to 26, characterized in that the second isolated monoclonal antibody is an immunoglobulin molecule comprising two heavy chains and two light chains linked by disulfide bonds, wherein each heavy chain comprises an HCVR comprising domains HCDR1, HCDR2 and HCDR3, and each light chain comprises an LCVR comprising domains LCDR1, LCDR2 and LCDR3.

29. Use of a pharmaceutical composition as defined in any one of claims 15 to 28, characterized in that it is for the manufacture of a medicament to treat or prevent EBOV infection in an individual in need thereof.

30. Use according to claim 29, characterized in that the composition comprises an antibody cocktail comprising a mixture of at least two anti-EBOV antibodies.

31. Use according to claim 29, characterized in that the medicament is formulated for prophylactic or therapeutic administration.

32. Use according to claim 29, characterized in that the individual in need thereof is an individual suffering from an EBOV infection or an individual exposed to or at risk of exposure to EBOV or of acquiring an EBOV infection, wherein the individual is selected from the group consisting of an immunocompromised individual, a health worker, a person suspected of having been exposed to a person carrying the Ebola virus, a person who comes into physical contact or close physical proximity to an infected individual, a hospital employee, a pharmaceutical researcher, maintenance staff responsible for cleaning a hospital facility or institution where an Ebola patient has been treated, individuals who have visited or are planning to visit an area or country known to have or suspected of having an Ebola virus outbreak, and a frequent airline passenger.

33. Use in accordance with any of claims 29 to 32, characterized in that the pharmaceutical composition is combined with a second therapeutic agent.

34. Use according to claim 33, characterized in that the second therapeutic agent is selected from the group consisting of an antiviral drug, an anti-inflammatory drug (e.g., corticosteroids and non-steroidal anti-inflammatory drugs), a different antibody for EBOV, an EBOV vaccine, TKM Ebola (small interfering RNAs targeting viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (pre-sense phosphorodiamidate morpholino oligomers targeting the VP24 gene of the Ebola virus) and interferons.

35. Use in accordance with any of claims 29 to 34, characterized in that the pharmaceutical composition is formulated for subcutaneous, intravenous, intradermal, intramuscular, intranasal or oral administration.

36. Use of the pharmaceutical composition as defined in any of claims 15 to 28, characterized in that it is for the manufacture of a medicament to increase the survival or probability of survival of an individual suffering from EBOV infection or an individual exposed to EBOV or at risk of exposure to, or acquisition of EBOV in an individual in need thereof.

37. Use according to claim 36, characterized in that the composition comprises an antibody cocktail comprising a mixture of at least two anti-EBOV antibodies.

38. Use in accordance with any of claims 36 or 37, characterized in that the individual in need thereof, who is at risk of being exposed to or acquiring an EBOV infection, is selected from the group consisting of an immunocompromised individual, a healthcare professional, a person suspected of having been exposed to a person carrying the Ebola virus, a person who comes into physical contact or close physical proximity to an infected individual, a hospital worker, a pharmaceutical researcher, maintenance staff responsible for cleaning a hospital facility or institution where an Ebola patient has been treated, individuals who have visited or are planning to visit an area or country known to have or suspected of having an Ebola virus outbreak, and a frequent airline passenger.

39. Use in accordance with any of claims 36 to 38, characterized in that the pharmaceutical composition or antibody cocktail is combined with a second therapeutic agent.

40. Use according to claim 39, characterized in that the second therapeutic agent is selected from the group consisting of an antiviral drug, an anti-inflammatory drug (e.g., corticosteroids and non-steroidal anti-inflammatory drugs), a different antibody for EBOV, an EBOV vaccine, TKM Ebola (small interfering RNAs targeting viral RNA polymerase), brincidofovir (CMX-001), favipiravir (T-705), BCX-4430, AVI-7537 (presensible phosphorodiamidate morpholino oligomers targeting the VP24 gene of the Ebola virus) and interferons.

41. Use in accordance with any of claims 36 to 40, characterized in that the pharmaceutical composition is formulated for subcutaneous, intravenous, intradermal, intramuscular, intranasal or oral administration.

42. Recombinant antibody isolated according to claim 1, characterized in that the antibody possesses one or more of the following features: (a) it is a fully human monoclonal antibody; (b) it binds to EBOV or a virus-like particle (VLP) expressing an EBOV-GP with a dissociation constant (Kd) less than 10⁻⁷ M, as measured in a surface plasma resonance assay; (c) it demonstrates at least a 2-fold increase in dissociative half-life (1½) at pH 5 or pH 6 compared to pH 7.4; (d) it demonstrates neutralization of Zaire Ebola virus with IC50 ranging from about 10⁻¹¹ M to about 10⁻⁹ M; and (e) it cross-reacts to one or more EBOV strains selected from the group consisting of Zaire.2014, Zaire.1995, Sudan, Bundibugyo and Cote d'Ivoire. Petition 870260058248, dated 06 / 15 / 2026, pp. 128 / 143

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