Patent Publication Number: US-11649276-B2

Title: Broadly-neutralizing anti-HIV antibodies

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a Divisional of application Ser. No. 16/507,867 filed on Jul. 10, 2019, which is a Divisional of U.S. patent application Ser. No. 16/006,420, filed Jun. 12, 2018, issued as U.S. Pat. No. 10,392,433 on Aug. 27, 2019, which is a Divisional of U.S. patent application Ser. No. 14/436,608, filed Apr. 17, 2015, issued as U.S. Pat. No. 10,047,146 on Aug. 14, 2018, which is the U.S. National Phase of International Application No. PCT/US2013/065696, filed Oct. 18, 2013, which claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 61/715,642 filed on Oct. 18, 2012, which are hereby incorporated by reference in their entireties. 
    
    
     GOVERNMENT INTERESTS 
     The invention disclosed herein was made, at least in part, with government support under Grant No. P01 AI081677 from the National Institutes of Health. Accordingly, the U.S. Government has certain rights in this invention. 
    
    
     FIELD OF THE INVENTION 
     This invention relates to broad and potent antibodies against Human Immunodeficiency Virus (“HIV”). 
     BACKGROUND OF THE INVENTION 
     HIV causes acquired immunodeficiency syndrome (AIDS), a condition in humans characterized by clinical features including wasting syndromes, central nervous system degeneration and profound immunosuppression that results in life-threatening opportunistic infections and malignancies. Since its discovery in 1981, HIV type 1 (HIV-1) has led to the death of at least 25 million people worldwide. It is predicted that 20-60 million people will become infected over the next two decades even if there is a 2.5% annual decrease in HIV infections. There is a need for therapeutic agents and methods for treatment or inhibition of HIV infection. 
     Some HIV infected individuals show broadly neutralizing IgG antibodies in their serum. Yet, little is known regarding the specificity and activity of these antibodies, despite their potential importance in designing effective vaccines. In animal models, passive transfer of neutralizing antibodies can contribute to protection against virus challenge. Neutralizing antibody responses also can be developed in HIV-infected individuals but the detailed composition of the serologic response is yet to be fully uncovered. 
     SUMMARY OF INVENTION 
     This invention relates to new categories of broadly-neutralizing anti-HIV antibodies. The consensus heavy and light chain amino acid sequences of the antibodies are listed below and shown in  FIGS.  3   a  and  3   b   : 
     
       
         
           
               
            
               
                 (SEQ ID NO: 1) 
               
               
                 QVQLQESGPGLVKPSETLSLICSVSGX 1 SX 2 X 3 DX 4 YWSWIRQSPGKGL 
               
               
                   
               
               
                 EWIGYVHDSGDTNYNPSLKSRVX 5 X 6 SLDTSKNQVSLKL X7 X 8 VTAADS 
               
               
                   
               
               
                 AX 9 YYCARAX 10 HGX II RTYGIVAFGEX 12 FTYFYMDVWGKGTTVTVSS 
               
               
                   
               
               
                 (SEQ ID NO: 2) 
               
               
                 SX 1 VRPQPPSLSVAPGETARIX 2 CGEX 3 SLGSRAVQWYQQRPGQAPSLI 
               
               
                   
               
               
                 TYNNQDRPSGIPERFSGSPDX 4 X 5 FGTTATLTITX 6 VEAGDEADYYCHI 
               
               
                   
               
               
                 WDSRX 7 PTX 8 WVFGGGTTLTVL 
               
            
           
         
       
     
     In the sequence of SEQ ID NO: 1 or 2, each “X” can be any amino acid residue or no amino acid. Preferably, each of the Xs can be a residue at the corresponding location of clonal variants 10-259, 10-303, 10-410, 10-847, 10-996, 10-1074, 10-1121, 10-1130, 10-1146, 10-1341, and 10-1369 as shown in  FIGS.  3   a  and  3   b   , and an artificially modified version of 10-1074 antibody, 10-1074GM. 
     Accordingly, one aspect of this invention features an isolated anti-HIV antibody, or antigen binding portion thereof, having at least one complementarity determining region (CDR) having a sequence selected from the group consisting of SEQ ID NOs: 33-38, with a proviso that the antibody is not antibody PGT-121, 122, or 123. SEQ ID NOs: 33-38 refer to the sequences of heavy chain CDRs (CDRH) 1-3 and the light chain CDRs (CDRL) 1-3 under the Kabat system as shown in  FIGS.  3   a  and  3   b   . In one embodiment, the CDR can contain a sequence selected from the group consisting of SEQ ID NOs: 39-104, i.e., the CDR sequences under the KABAT system as shown in Table 1 below. Alternatively, the CDR can contain a sequence selected from those corresponding antibodies&#39; CDR sequences under the IMGT system as shown in Table 1 below. 
     In one embodiment, the isolated anti-HIV antibody, or antigen binding portion thereof, contains a heavy chain variable region that includes CDRH 1, CDRH 2, and CDRH 3, wherein the CDRH 1, CDRH 2 and CDRH 3 include the respective sequences of SEQ ID NOs: 33-35. The CDRH 1, CDRH 2 and CDRH 3 can also include the respective sequences of a CDRH set selected from the group consisting of SEQ ID NOs: 39-41, SEQ ID NOs: 45-47, SEQ ID NOs: 51-53, SEQ ID NOs: 57-59, SEQ ID NOs: 63-65, SEQ ID NOs: 69-71, SEQ ID NOs: 75-77, SEQ ID NOs: 81-83, SEQ ID NOs: 87-89, SEQ ID NOs: 93-95, SEQ ID NOs: 99-101, and SEQ ID NOs: 131-133. Alternatively, the CDRHs can contain the respective sequences selected from those corresponding antibodies&#39; CDR sequences under the IMGT system as shown in Table 1 below. 
     In another embodiment, the isolated anti-HIV antibody, or antigen binding portion thereof, contains a light chain variable region that includes CDRL 1, CDRL 2 and CDRL 3, wherein the CDRL 1, CDRL 2 and CDRL 3 include the respective sequences of SEQ ID NOs: 36-38. For example, the CDRL 1, CDRL 2 and CDRL 3 can include the respective sequences of a CDRL set selected from the group consisting of SEQ ID NOs: 42-44, SEQ ID NOs: 48-50, SEQ ID NOs: 54-56, SEQ ID NOs: 60-62, SEQ ID NOs: 66-68, SEQ ID NOs: 72-74, SEQ ID NOs: 78-80, SEQ ID NOs: 84-86, SEQ ID NOs: 90-92, SEQ ID NOs: 96-98, SEQ ID NOs: 102-104, and SEQ ID NOs: 134-136. Alternatively, the CDRLs can contain the respective sequences selected from those corresponding antibodies&#39; CDR sequences under the IMGT system as shown in Table 1 below. 
     In yet another embodiment, the above-mentioned isolated anti-HIV antibody, or antigen binding portion thereof, includes (i) a heavy chain variable region that include CDRH 1, CDRH 2, and CDRH 3, and (ii) a light chain variable region that include CDRL 1, CDRL 2 and CDRL 3. The CDRH 1, CDRH 2, CDRH 3, CDRL 1, CDRL 2 and CDRL 3 can include the respective sequences of a CDR set selected from the group consisting of SEQ ID NOs: 39-44, SEQ ID NOs: 45-50, SEQ ID NOs: 51-56, SEQ ID NOs: 57-62, SEQ ID NOs: 63-68, SEQ ID NOs: 69-74, SEQ ID NOs: 75-79, SEQ ID NOs: 81-86, SEQ ID NOs: 87-92, SEQ ID NOs: 93-98, SEQ ID NOs: 99-104, and SEQ ID NOs: 131-136. Alternatively, the CDRHs and CDRLs can contain the respective sequences selected from those corresponding antibodies&#39; CDR sequences under the IMGT system as shown in Table 1 below. 
     In a further embodiment, the isolated anti-HIV antibody, or antigen binding portion thereof, contains one or both of (i) a heavy chain having the consensus amino acid sequence of SEQ ID NO: 1 and (ii) a light chain having the consensus amino acid sequence of SEQ ID NO: 2. The heavy chain can contain a sequence selected from the group consisting of SEQ ID NOs: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, and 129, and the light chain can contain a sequence selected from the group consisting of SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, and 130. For example, the heavy chain and the light chain can include the respective sequences of SEQ ID NOs: 3-4, SEQ ID NOs: 5-6, SEQ ID NOs: 7-8, SEQ ID NOs: 9-10, SEQ ID NOs: 11-12, SEQ ID NOs: 13-14, SEQ ID NOs: 15-16, SEQ ID NOs: 17-18, SEQ ID NOs: 19-20, SEQ ID NOs: 21-22, SEQ ID NOs: 23-24, and 129-130. 
     In a preferred embodiment, the isolated anti-HIV antibody is one selected from the group consisting of 10-259, 10-303, 10-410, 10-847, 10-996, 10-1074, 10-1074GM, 10-1121, 10-1130, 10-1146, 10-1341, and 10-1369. Their corresponding heavy chain variable regions, light chain variable regions, CDRH 1-3 and CDRL 1-3 are shown in  FIGS.  3   a  and  3   b   . In a more preferred embodiment, the isolated anti-HIV antibody is a 10-1074-like antibody, i.e., one reselected from the group consisting of 10-847, 10-996, 10-1074, 10-1074GM, 10-1146, and 10-1341. An antibody of this group is more potent in neutralizing contemporary viruses than PGT121. The above-discussed antibody can be a human antibody, a humanized antibody, or a chimeric antibody. 
     In a second aspect, the invention provides an isolated nucleic acid having a sequence encoding a CDR, a heavy chain variable region, or a light chain variable region of the above-discussed anti-HIV antibody, or antigen binding portion thereof. Also featured are a vector having the nucleic acid and a cultured cell having the vector. 
     The nucleic acid, vector, and cultured cell can be used in a method for making an anti-HIV antibody or a fragment thereof. The method includes, among others, the steps of: obtaining the cultured cell mentioned above; culturing the cell in a medium under conditions permitting expression of a polypeptide encoded by the vector and assembling of an antibody or fragment thereof, and purifying the antibody or fragment from the cultured cell or the medium of the cell. 
     In a third aspect, the invention features a pharmaceutical composition containing (i) at least one anti-HIV antibody mentioned above, or antigen binding portion thereof, and (ii) a pharmaceutically acceptable carrier. 
     In a fourth aspect, the invention provides a method of preventing or treating an HIV infection or an HIV-related disease. The method includes, among others, the steps of: identifying a patient in need of such prevention or treatment, and administering to said patient a first therapeutic agent containing a therapeutically effective amount of at least one anti-HIV antibody mentioned above, or antigen binding portion thereof. The method can further include administering a second therapeutic agent, such as an antiviral agent. 
     In a fifth aspect, the invention provides a kit having a pharmaceutically acceptable dose unit of a pharmaceutically effective amount of at least one isolated anti-HIV antibody mentioned a above, or antigen binding portion thereof, and a pharmaceutically acceptable dose unit of a pharmaceutically effective amount of an anti-HIV agent. The two pharmaceutically acceptable dose units can optionally take the form of a single pharmaceutically acceptable dose unit. Exemplary anti-HIV agent can be one selected from the group consisting of a non-nucleoside reverse transcriptase inhibitor, a protease inhibitor, a entry or fusion inhibitor, and an integrase inhibitor. 
     In a sixth aspect, the invention provides a kit for the diagnosis, prognosis or monitoring the treatment of an HIV infection in a subject. The kit contains one or more detection reagents which specifically bind to anti-HIV neutralizing antibodies in a biological sample from a subject. The kit can further include reagents for performing PCR or mass spectrometry. 
     The details of one or more embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will be apparent from the description and from the claims. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIGS.  1 A,  1 B,  1 C,  1 D and  1 E  show: Neutralization activity of PGT121-like and 10-1074-like variants. (A) Heat map comparing the neutralization potencies of PGT121-like and 10-1074-like antibodies in the TZM-bl assay. Darker colors=more potent neutralization; white=no neutralization. (B) Correlation between the mean IC 80  against 9 viruses (y axis) and apparent K D  values for binding to gp120 and gp140 (x axis). (C) Graph comparing the neutralization breadth and potencies of PGT121, 10-996 and 10-1074 antibodies in the TZM-bl assay against an extended panel of 119 viruses. The y axis shows the cumulative frequency of IC 50  values up to the concentration shown on the x axis. The spider graph (upper left corner) shows the frequency distribution of neutralized viruses according to HIV-1 clades. (D) Dot plot showing molar neutralization ratios (MNRs; ratio of the Fab and IgG IC 50  concentrations). Horizontal bars represent the mean IC 50 s for all viruses. (E) Bar graph comparing the neutralization potencies of PGT121 (dark gray) and 10-1074 (light gray) against viruses isolated from historical (Hist.) and contemporary (Cont.) seroconverters. ns, non significant; **, p&lt;0.005. Fold difference between median IC 50 s for the neutralization of contemporary viruses by PGT121 and 10-1074 is indicated. 
         FIGS.  2 A,  2 B and  2 C  show: Binding and neutralization activities of PGT121 GM  and 10-1074 GM  mutant antibodies. (A) Bar graphs comparing apparent K D  values for the binding of 10-1074, PGT121, PGT121 GM  and 10-1074 GM  antibodies to gp120 and gp140. Error bars indicate the SEM of K D  values obtained from three independent experiments. Fold differences between K D  values of “wildtype” vs “glycomutant” antibodies are indicated. (B) Bar graphs comparing binding of glycans ( FIG.  7 A ) by PGT121 and 10-1074 with mutant antibodies (PGT121 GM  and 10-1074 GM ). Numerical scores of binding are measured as fluorescence intensity (means at duplicate spots) for probes arrayed at 5 fmol per spot. (C) Coverage graph comparing the neutralization breadth and potencies of PGT121, PGT121 GM , 10-1074 and 10-1074 GM  antibodies in the TZM-bl assay against a panel of 40 viruses. 
         FIGS.  3 A and  3 B  depict: Sequence alignments of PGT121 and 10-1074 clonal variants. (A) Amino acid alignment of the heavy chains (IgH) of the PGT121-like and 10-1074-like antibodies, and the likely germline (GL) VH for all clonal variants. Amino acid numbering based on crystal structures, framework (FWR) and complementary determining regions (CDR) as defined by Kabat (J Exp Med 132(2):211-250) and IMGT (Nucleic Acids Res 37 (Database issue):D1006-1012) are indicated. Color shading shows acidic (red), basic (blue), and tyrosine (green) amino acids. (B) Same as A but for the light chains (IgL).  FIG.  3 A  discloses SEQ ID NOS 31, 9, 21, 13, 19, 11, 23, 27, 3, 29, 17, 15, 7, 25, 5, and 1, respectively, in order of appearance.  FIG.  3 B  discloses SEQ ID NOS 32, 26, 6, 24, 30, 4, 10, 14, 22, 12, 20, 8, 28, 18, 16, and 2, respectively, in order of appearance. 
         FIGS.  4 A,  4 B and  4 C  show: Binding affinity of PGT121 and 10-1074 clonal variants. (A) Binding affinity of the interaction of PGT121 IgG antibody variants with YU-2 gp140 and gp120 ligands as measured by surface plasmon resonance (SPR). M, mol/l; s, seconds; RU, response units; /, no binding detected. A chi 2  value (χ 2 )&lt;10 indicates that the 1:1 binding model used to fit the curves adequately described the experimental data. Equilibrium and kinetic constants shown are considered as “apparent” constants to account for avidity effects resulting from bivalent binding of IgGs. (B) Dot plots showing the association (k a ) and dissociation (k d ) rate constants for PGT121-like (blue shading) and 10-1074-like (green shading). (C) Linear regression graphs comparing the k a  and k d  values of the IgG antibodies for their binding to gp120 and gp140 (x axis) vs their neutralization potencies (mean IC 80  values) against the 9 viruses shown in Table 4 (y axis). 
         FIGS.  5 A,  5 B and  5 C  depict: Binding of PGT121 variants to gp120 “core” proteins, gp120 GD324-5AA  mutant and linear gp120 V3  peptides. (A) ELISA-based binding analyses of PGT121-like and 10-1074-like antibodies to HXB2 gp120 core  and 2CC-core proteins compared to intact YU-2 gp120. The x axis shows the antibody concentration (M) required to obtain the ELISA values (OD 405 nm ) indicated on the y axis. The anti-CD4bs antibody VRC01 (Science 329(5993):856-861), the anti-V3 loop antibody 10-188 (PLoS One 6(9):e24078), and the non HIV-reactive antibody mGO53 (Science 301(5638):1374-1377) were used as controls. (B) Same as (A) but for binding to gp120 GD324-5AA  mutant protein (c) Bar graphs comparing the ELISA reactivities of the PGT121- and 10-1074-like antibodies and control antibodies (positive control, 10-188, 1-79, 2-59 and 2-1261 (Nature 458(7238):636-640)), and negative control, mGO53) against gp120 V3-C3  overlapping peptides. The y axis indicates the ELISA values (OD 405 nm ) obtained by testing the IgG antibodies at 2 μg/ml. The amino acid sequences of individual peptides are shown in the bottom right. All experiments were performed at least in duplicate. Representative data are shown.  FIG.  5 C  discloses SEQ ID NOS 216-223, respectively, in order of appearance. 
         FIGS.  6 A,  6 B,  6 C and  6 D  depict: Binding of PGT121 to gp120 glycosylation mutants and deglycosylated gp120. (A) ELISA-based binding analyses of PGT121 and 10-1074 antibody variants to gp120, gp120 NNT301-303AAA , gp120 N332A  and gp120 N332A/NNT301-303AAA . The x axis shows the antibody concentration (M) required to obtain the ELISA values (OD 405 nm ) indicated on the y axis. The black dashed and continuous lines show the averaged reactivity against the four antigens of positive (10-188) and negative (mGO53) antibody controls. (B) Silver-stained SDS-PAGE gel comparing untreated gp120 (WT, wild type), PNGase F- and EndoH-digested gp120s. L, protein ladder. (C), Same as (A) but comparing untreated and PNGase F-treated gp120. (D) Same as (A) but comparing untreated and EndoH-treated gp120. All experiments were performed at least in duplicate. 
         FIGS.  7 A and  7 B  depict: Binding of PGT121 and 10-1074 clonal variants to glycans. ( FIG.  7 A ) Monosaccharide sequences of the set of 15 N-glycan probes used in the glycan microarray analyses to examine PGT121-like and 10-1074-like antibodies for direct binding to N-glycans. DH, designates the lipid tag 1,2-dihexadecyl-sn-glycero-3-phosphoethanolamine (DHPE) to which the N-glycans were conjugated by reductive amination. Key features of note are (i) PGT121-group antibodies bound the monoantennary N-glycan probe 10 (N2) with a galactose-terminating antenna joined by 1-3-linkage to the core mannose, but not the isomeric N-glycan probe 11 (designated N4) with the antenna 1-6-linked to the core mannose; (ii) the presence of this galactose-terminating 1-6-linked antenna, as in the biantennary probe 13 (NA2), was permissive to binding, as was the presence of α2-6-linked (but not α2-3-linked) sialic acid; (iii) the biantennary probe 12 (NGA2), lacking galactose and terminating in N-acetylglucosamine, was not bound. ( FIG.  7 B ) Bar graphs comparing glycan binding by PGT121-like, 10-1074-like, and the germline version (GL) antibodies. 10-188, an anti-V3 loop antibody, was used as negative control. Numerical scores of binding are measured as fluorescence intensity (means at duplicate spots) for probes arrayed at 2 fmol (white) and 5 fmol per spot (grey). 
         FIGS.  8 A,  8 B,  8 C and  8 D  depict: Antibody binding and neutralization activity against high-mannose-only gp120 and viruses. (A) Silver-stained SDS-PAGE gel comparing YU-2 gp120 produced in cells treated with kifunensine (gp120 kif ) and gp120 produced in untreated cells (WT, wild type). L, protein ladder. (B) ELISA comparison of the binding of PGT121-like (blue labels) and 10-1074-like (green labels) antibodies to YU-2 gp120 (gp120 WT ) and gp120 kif . The x axis shows the antibody concentration (M) required to obtain the ELISA values (OD 405 nm ) indicated on they axis. (C) Neutralization curves for PGT121 evaluated against selected PGT121-sensitive/10-1074-resistant pseudoviruses produced in presence (Virus kif ) or absence (Virus WT ) of kifunensine. The dotted horizontal line indicates 50% neutralization, from which the IC 50  value can be derived from the antibody concentration on the x axis. Experiments were performed in triplicate. Error bars indicate the SD of triplicate measurements. (D) Bar graphs comparing the neutralization activity of selected antibodies against YU-2 and PVO.4 pseudoviruses produced in HEK 293S GnTI −/−  cells (Virus GnT   −/− ) or in wild type cells (Virus WT ). They axis shows the mean IC 50  values (μg/ml) for the neutralization of the viruses shown on the x axis. Error bars indicate the SEM of IC 50  values obtained from two independent experiments. 
         FIGS.  9 A,  9 B and  9 C  show: Neutralization activity of PGT121, 10-996 and 10-1074. (A) Graphs comparing the neutralization potencies of PGT121, 10-996 and 10-74 against viruses of the indicated HIV-1 clades (determined using the TZM-bl assay and a panel of 119 pseudoviruses). The x axis shows the antibody concentration (m/ml) required to achieve 50% neutralization (IC 50 ). The y axis shows the cumulative frequency of IC 50  values up to the concentration shown on the x axis. (B) Graph comparing the neutralization breadth and potencies of PGT121, 10-996 and 10-1074 antibodies against the extended panel of 119 viruses as determined by the TZM-bl neutralization assay. The y axis shows the cumulative frequency of IC 50  values up to the concentration shown on the x axis. (C) Graphs show neutralization curves of the selected viruses by PGT121 and 10-1074. The dotted horizontal line indicates 50% neutralization, from which the IC 50  value can be derived from the antibody concentration on the x-axis. Experiments were performed in triplicate. Error bars indicate the SD of triplicate measurements. 
         FIG.  10    depicts: Neutralization activity against historical vs contemporary clade B viruses. Dot plots comparing neutralization potencies against clade B viruses isolated from historical (Hist.) and contemporary (Cont.) seroconverters for the selected bNAbs. Horizontal bars represent the median IC 50  for all viruses per patient. Differences between groups were evaluated using Mann-Whitney test. ns, not significant. 
         FIGS.  11 A and  11 B  depict: Neutralization of two R5 tropic SHIVs with a panel of 11 broadly acting anti-HIV-1 mAbs. The calculated IC50 values for neutralizing SHIVAD8EO (A) and SHIVDH12-V3AD8 (B). 
         FIG.  12    depicts: The relationship of the plasma concentrations of passively administered neutralizing mAbs to virus acquisition following challenge of macaques with two different R5 SHIVs. Filled circles indicate protected (no acquisition) monkeys; open circles denote infected animals. 
         FIGS.  13 A,  13 B and  13 C  depict: Plasma concentration of bNAbs. The concentration of mAbs was determined by measuring neutralization activity in plasma samples. (A) ID50-values measured in TZM.b1 neutralization assay of 10-1074 and 3BNC117 against HIV-1 strains that are sensitive to one but not the other bNAb (i.e. HIV-1 strain X2088_9 (10-1074 sensitive); HIV-1 strain Q769_d22 (3BNC117 sensitive). (B) Neutralizing activity of plasma before antibody administration (preP), but spiked with 0.01, 0.1, 1, 10, and 100 μg/ml of antibodies 10-1074 (blue) or 3BNC117 (green). Neutralizing activity reported as plasma ID50 titers (left columns) and converted to antibody concentrations (right columns) based on measured ID50-values in (A). (C) ID50 titers (left columns) and concentrations of bNAbs (right columns) measured in the indicated macaque plasma samples before (Prebleed) and following (Day) bNAb administration. 
         FIG.  14    depicts Table 7 showing neutralization sensitivity according to N332 PNGS. Dots indicate the presence of an asparagine and of a serine residue in position 332 and 334, respectively. Mutations at positions 332 and 334 (HXB2 sequence number) are indicated by the substituting amino acid. IC50 values indicate neutralization sensitivity in the TZM-bl assay. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     This invention is based, at least in part, on an unexpected discovery of a new category of broadly neutralizing antibodies (bNAbs) against HIV that can recognize carbohydrate-dependent epitopes, including complex-type N-glycan, on gp120. 
     Antibodies are essential for the success of most vaccines, and antibodies against HIV appear to be the only correlate of protection in the recent RV144 anti-HIV vaccine trial. Some HIV-1 infected patients develop broadly neutralizing serologic activity against the gp160 viral spike 2-4 years after infection, but these antibodies do not generally protect infected humans because autologous viruses escape through mutation. Nevertheless, broadly neutralizing activity puts selective pressure on the virus and passive transfer of broadly neutralizing antibodies (bNAbs) to macaques protects against SHIV infection. It has therefore been proposed that vaccines that elicit such antibodies may be protective against HIV infection in humans. 
     The development of single cell antibody cloning techniques revealed that bNAbs target several different epitopes on the HIV-1 gp160 spike. The most potent HIV-1 bNAbs recognize the CD4 binding site (CD4bs) (Science 333(6049):1633-1637; Nature 477(7365):466-470; Science 334(6060):1289-1293) and carbohydrate-dependent epitopes associated with the variable loops (Nature 477(7365):466-470; Science 326(5950):285-289; Science 334(6059):1097-1103; Nature 480(7377):336-343), including the V1/V2 (PG9/PG16) (Science 326(5950):285-289) and V3 loops (PGTs) (Nature 477(7365):466-470). Less is known about carbohydrate-dependent epitopes because the antibodies studied to date are either unique examples or members of small clonal families. 
     To better understand the neutralizing antibody response to HIV-1 and the epitope targeted by PGT antibodies, we isolated members of a large clonal family dominating the gp160-specific IgG memory response from the clade A-infected patient who produced PGT121. As disclosed herein, PGT121 antibodies segregate into two groups, a PGT121-like and a 10-1074-like group, according to sequence, binding affinity, neutralizing activity and recognition of carbohydrates and the V3 loop. 10-1074 and related family members exhibit unusual potent neutralization, including broad reactivity against newly-transmitted viruses. Unlike previously-characterized carbohydrate-dependent bNAbs, PGT121 binds to complex-type, rather than high-mannose, N-glycans in glycan microarray experiments. Crystal structures of PGT121 and 10-1074 compared with structures of their germline precursor and a structure of PGT121 bound to a complex-type N-glycan rationalize their distinct properties. 
     In one example, assays were carried out to isolate B-cell clones encoding PGT121, which is unique among glycan-dependent bNAbs in recognizing complex-type, rather than high-mannose, N-glycans. The PGT121 clones segregates into PGT121- and 10-1074-like groups distinguished by sequence, binding affinity, carbohydrate recognition and neutralizing activity. The 10-1074 group exhibit remarkable potency and breadth despite not binding detectably to protein-free glycans. Crystal structures of un-liganded PGT121, 10-1074, and their germline precursor reveal that differential carbohydrate recognition maps to a cleft between CDRH2 and CDRH3, which was occupied by a complex-type N-glycan in a separate PGT121 structure. Swapping glycan contact residues between PGT121 and 10-1074 confirmed the importance of these residues in neutralizing activities. HIV envelopes exhibit varying proportions of high-mannose- and complex-type N-glycans, thus these results, including the first structural characterization of complex-type N-glycan recognition by anti-HIV bNAbs, are critical for understanding how antibodies and ultimately vaccines might achieve broad neutralizing activity. 
     The term “antibody” (Ab) as used herein includes monoclonal antibodies, polyclonal antibodies, multispecific antibodies (for example, bispecific antibodies and polyreactive antibodies), and antibody fragments. Thus, the term “antibody” as used in any context within this specification is meant to include, but not be limited to, any specific binding member, immunoglobulin class and/or isotype (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA, IgD, IgE and IgM); and biologically relevant fragment or specific binding member thereof, including but not limited to Fab, F(ab′)2, Fv, and scFv (single chain or related entity). It is understood in the art that an antibody is a glycoprotein having at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, or an antigen binding portion thereof. A heavy chain is comprised of a heavy chain variable region (VH) and a heavy chain constant region (CH1, CH2 and CH3). A light chain is comprised of a light chain variable region (VL) and a light chain constant region (CL). The variable regions of both the heavy and light chains comprise framework regions (FWR) and complementarity determining regions (CDR). The four FWR regions are relatively conserved while CDR regions (CDR1, CDR2 and CDR3) represent hypervariable regions and are arranged from NH2 terminus to the COOH terminus as follows: FWR1, CDR1, FWR2, CDR2, FWR3, CDR3, and FWR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen while, depending of the isotype, the constant region(s) may mediate the binding of the immunoglobulin to host tissues or factors. 
     Also included in the definition of “antibody” as used herein are chimeric antibodies, humanized antibodies, and recombinant antibodies, human antibodies generated from a transgenic non-human animal, as well as antibodies selected from libraries using enrichment technologies available to the artisan. 
     The term “variable” refers to the fact that certain segments of the variable (V) domains differ extensively in sequence among antibodies. The V domain mediates antigen binding and defines specificity of a particular antibody for its particular antigen. However, the variability is not evenly distributed across the 110-amino acid span of the variable regions. Instead, the V regions consist of relatively invariant stretches called framework regions (FRs) of 15-30 amino acids separated by shorter regions of extreme variability called “hypervariable regions” that are each 9-12 amino acids long. The variable regions of native heavy and light chains each comprise four FRs, largely adopting a beta sheet configuration, connected by three hypervariable regions, which form loops connecting, and in some cases forming part of, the beta sheet structure. The hypervariable regions in each chain are held together in close proximity by the FRs and, with the hypervariable regions from the other chain, contribute to the formation of the antigen-binding site of antibodies (see, for example, Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). 
     The term “hypervariable region” as used herein refers to the amino acid residues of an antibody that are responsible for antigen binding. The hypervariable region generally comprises amino acid residues from a “complementarity determining region” (“CDR”). 
     The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. The term “polyclonal antibody” refers to preparations that include different antibodies directed against different determinants (“epitopes”). 
     The monoclonal antibodies herein include “chimeric” antibodies in which a portion of the heavy and/or light chain is identical with, or homologous to, corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with, or homologous to, corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (see, for example, U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). The described invention provides variable region antigen-binding sequences derived from human antibodies. Accordingly, chimeric antibodies of primary interest herein include antibodies having one or more human antigen binding sequences (for example, CDRs) and containing one or more sequences derived from a non-human antibody, for example, an FR or C region sequence. In addition, chimeric antibodies included herein are those comprising a human variable region antigen binding sequence of one antibody class or subclass and another sequence, for example, FR or C region sequence, derived from another antibody class or subclass. 
     A “humanized antibody” generally is considered to be a human antibody that has one or more amino acid residues introduced into it from a source that is non-human. These non-human amino acid residues often are referred to as “import” residues, which typically are taken from an “import” variable region. Humanization may be performed following the method of Winter and co-workers (see, for example, Jones et al., Nature, 321:522-525 (1986); Reichmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by substituting import hypervariable region sequences for the corresponding sequences of a human antibody. Accordingly, such “humanized” antibodies are chimeric antibodies (see, for example, U.S. Pat. No. 4,816,567), where substantially less than an intact human variable region has been substituted by the corresponding sequence from a non-human species. 
     An “antibody fragment” comprises a portion of an intact antibody, such as the antigen binding or variable region of the intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab′, F(ab′)2, and Fv fragments; diabodies; linear antibodies (see, for example, U.S. Pat. No. 5,641,870; Zapata et al., Protein Eng. 8(10): 1057-1062 [1995]); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. 
     “Fv” is the minimum antibody fragment that contains a complete antigen-recognition and antigen-binding site. This fragment contains a dimer of one heavy- and one light-chain variable region domain in tight, non-covalent association. From the folding of these two domains emanate six hypervariable loops (three loops each from the H and L chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody. However, even a single variable region (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site. 
     “Single-chain Fv” (“sFv” or “scFv”) are antibody fragments that comprise the VH and VL antibody domains connected into a single polypeptide chain. The sFv polypeptide can further comprise a polypeptide linker between the VH and VL domains that enables the sFv to form the desired structure for antigen binding. For a review of sFv, see, for example, Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994); Borrebaeck 1995, infra. 
     The term “diabodies” refers to small antibody fragments prepared by constructing sFv fragments with short linkers (about 5-10 residues) between the VH and VL domains such that inter-chain but not intra-chain pairing of the V domains is achieved, resulting in a bivalent fragment, i.e., fragment having two antigen-binding sites. Bispecific diabodies are heterodimers of two “crossover” sFv fragments in which the VH and VL domains of the two antibodies are present on different polypeptide chains. Diabodies are described more fully in, for example, EP 404,097; WO 93/11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993). 
     Domain antibodies (dAbs), which can be produced in fully human form, are the smallest known antigen-binding fragments of antibodies, ranging from about 11 kDa to about 15 kDa. DAbs are the robust variable regions of the heavy and light chains of immunoglobulins (VH and VL, respectively). They are highly expressed in microbial cell culture, show favorable biophysical properties including, for example, but not limited to, solubility and temperature stability, and are well suited to selection and affinity maturation by in vitro selection systems such as, for example, phage display. DAbs are bioactive as monomers and, owing to their small size and inherent stability, can be formatted into larger molecules to create drugs with prolonged serum half-lives or other pharmacological activities. Examples of this technology have been described in, for example, WO9425591 for antibodies derived from Camelidae heavy chain Ig, as well in US20030130496 describing the isolation of single domain fully human antibodies from phage libraries. 
     Fv and sFv are the only species with intact combining sites that are devoid of constant regions. Thus, they are suitable for reduced nonspecific binding during in vivo use. sFv fusion proteins can be constructed to yield fusion of an effector protein at either the amino or the carboxy terminus of an sFv. See, for example, Antibody Engineering, ed. Borrebaeck, supra. The antibody fragment also can be a “linear antibody”, for example, as described in U.S. Pat. No. 5,641,870 for example. Such linear antibody fragments can be monospecific or bispecific. 
     In certain embodiments, antibodies of the described invention are bispecific or multispecific. Bispecific antibodies are antibodies that have binding specificities for at least two different epitopes. Exemplary bispecific antibodies can bind to two different epitopes of a single antigen. Other such antibodies can combine a first antigen binding site with a binding site for a second antigen. Alternatively, an anti-HIV arm can be combined with an arm that binds to a triggering molecule on a leukocyte, such as a T-cell receptor molecule (for example, CD3), or Fc receptors for IgG (Fc gamma R), such as Fc gamma RI (CD64), Fc gamma RII (CD32) and Fc gamma RIII (CD16), so as to focus and localize cellular defense mechanisms to the infected cell. Bispecific antibodies also can be used to localize cytotoxic agents to infected cells. Bispecific antibodies can be prepared as full length antibodies or antibody fragments (for example, F(ab′)2 bispecific antibodies). For example, WO 96/16673 describes a bispecific anti-ErbB 2 /anti-Fc gamma RIII antibody and U.S. Pat. No. 5,837,234 discloses a bispecific anti-ErbB 2 /anti-Fc gamma RI antibody. For example, a bispecific anti-ErbB2/Fc alpha antibody is reported in WO98/02463; U.S. Pat. No. 5,821,337 teaches a bispecific anti-ErbB2/anti-CD3 antibody. See also, for example, Mouquet et al., Polyreactivity Increases The Apparent Affinity Of Anti-HIV Antibodies By Heteroligation. Nature. 467, 591-5 (2010), and Mouquet et al., Enhanced HIV-1 neutralization by antibody heteroligation” Proc Natl Acad Sci USA. 2012 Jan. 17; 109(3):875-80. 
     Methods for making bispecific antibodies are known in the art. Traditional production of full length bispecific antibodies is based on the co-expression of two immunoglobulin heavy chain-light chain pairs, where the two chains have different specificities (see, for example, Millstein et al., Nature, 305:537-539 (1983)). Similar procedures are disclosed in, for example, WO 93/08829, Traunecker et al., EMBO J., 10:3655-3659 (1991) and see also Mouquet et al., Enhanced HIV-1 neutralization by antibody heteroligation” Proc Natl Acad Sci USA. 2012 Jan. 17; 109(3):875-80. 
     Alternatively, antibody variable regions with the desired binding specificities (antibody-antigen combining sites) are fused to immunoglobulin constant domain sequences. The fusion is with an Ig heavy chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. According to some embodiments, the first heavy-chain constant region (CH1) containing the site necessary for light chain bonding, is present in at least one of the fusions. DNAs encoding the immunoglobulin heavy chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host cell. This provides for greater flexibility in adjusting the mutual proportions of the three polypeptide fragments in embodiments when unequal ratios of the three polypeptide chains used in the construction provide the optimum yield of the desired bispecific antibody. It is, however, possible to insert the coding sequences for two or all three polypeptide chains into a single expression vector when the expression of at least two polypeptide chains in equal ratios results in high yields or when the ratios have no significant affect on the yield of the desired chain combination. 
     Techniques for generating bispecific antibodies from antibody fragments also have been described in the literature. For example, bispecific antibodies can be prepared using chemical linkage. For example, Brennan et al., Science, 229: 81 (1985) describe a procedure wherein intact antibodies are proteolytically cleaved to generate F(ab′)2 fragments. These fragments are reduced in the presence of the dithiol complexing agent, sodium arsenite, to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The Fab′ fragments generated then are converted to thionitrobenzoate (TNB) derivatives. One of the Fab′-TNB derivatives then is reconverted to the Fab′-thiol by reduction with mercaptoethylamine and is mixed with an equimolar amount of the other Fab′-TNB derivative to form the bispecific antibody. The bispecific antibodies produced can be used as agents for the selective immobilization of enzymes. 
     Other modifications of the antibody are contemplated herein. For example, the antibody can be linked to one of a variety of nonproteinaceous polymers, for example, polyethylene glycol, polypropylene glycol, polyoxyalkylenes, or copolymers of polyethylene glycol and polypropylene glycol. The antibody also can be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively), in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules), or in macroemulsions. Such techniques are disclosed in, for example, Remington&#39;s Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980). 
     Typically, the antibodies of the described invention are produced recombinantly, using vectors and methods available in the art. Human antibodies also can be generated by in vitro activated B cells (see, for example, U.S. Pat. Nos. 5,567,610 and 5,229,275). General methods in molecular genetics and genetic engineering useful in the present invention are described in the current editions of Molecular Cloning: A Laboratory Manual (Sambrook, et al., 1989, Cold Spring Harbor Laboratory Press), Gene Expression Technology (Methods in Enzymology, Vol. 185, edited by D. Goeddel, 1991. Academic Press, San Diego, Calif.), “Guide to Protein Purification” in Methods in Enzymology (M. P. Deutshcer, ed., (1990) Academic Press, Inc.); PCR Protocols: A Guide to Methods and Applications (Innis, et al. 1990. Academic Press, San Diego, Calif.), Culture of Animal Cells: A Manual of Basic Technique, 2nd Ed. (R. I. Freshney. 1987. Liss, Inc. New York, N.Y.), and Gene Transfer and Expression Protocols, pp. 109-128, ed. E. J. Murray, The Humana Press Inc., Clifton, N.J.). Reagents, cloning vectors, and kits for genetic manipulation are available from commercial vendors such as BioRad, Stratagene, Invitrogen, ClonTech and Sigma-Aldrich Co. 
     Human antibodies also can be produced in transgenic animals (for example, mice) that are capable of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production. For example, it has been described that the homozygous deletion of the antibody heavy-chain joining region (JH) gene in chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production. Transfer of the human germ-line immunoglobulin gene array into such germ-line mutant mice results in the production of human antibodies upon antigen challenge. See, for example, Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immuno., 7:33 (1993); U.S. Pat. Nos. 5,545,806, 5,569,825, 5,591,669 (all of GenPharm); U.S. Pat. No. 5,545,807; and WO 97/17852. Such animals can be genetically engineered to produce human antibodies comprising a polypeptide of the described invention. 
     Various techniques have been developed for the production of antibody fragments. Traditionally, these fragments were derived via proteolytic digestion of intact antibodies (see, for example, Morimoto et al., Journal of Biochemical and Biophysical Methods 24:107-117 (1992); and Brennan et al., Science, 229:81 (1985)). However, these fragments can now be produced directly by recombinant host cells. Fab, Fv and ScFv antibody fragments can all be expressed in and secreted from  E. coli , thus allowing the facile production of large amounts of these fragments. Fab′-SH fragments can be directly recovered from  E. coli  and chemically coupled to form F(ab′)2 fragments (see, for example, Carter et al., Bio/Technology 10:163-167 (1992)). According to another approach, F(ab′)2 fragments can be isolated directly from recombinant host cell culture. Fab and F(ab′)2 fragment with increased in vivo half-life comprising a salvage receptor binding epitope residues are described in U.S. Pat. No. 5,869,046. Other techniques for the production of antibody fragments will be apparent to the skilled practitioner. 
     Other techniques that are known in the art for the selection of antibody fragments from libraries using enrichment technologies, including but not limited to phage display, ribosome display (Hanes and Pluckthun, 1997 , Proc. Nat. Acad. Sci.  94: 4937-4942), bacterial display (Georgiou, et al., 1997 , Nature Biotechnology  15: 29-34) and/or yeast display (Kieke, et al., 1997 , Protein Engineering  10: 1303-1310) may be utilized as alternatives to previously discussed technologies to select single chain antibodies. Single-chain antibodies are selected from a library of single chain antibodies produced directly utilizing filamentous phage technology. Phage display technology is known in the art (e.g., see technology from Cambridge Antibody Technology (CAT)) as disclosed in U.S. Pat. Nos. 5,565,332; 5,733,743; 5,871,907; 5,872,215; 5,885,793; 5,962,255; 6,140,471; 6,225,447; 6,291,650; 6,492,160; 6,521,404; 6,544,731; 6,555,313; 6,582,915; 6,593,081, as well as other U.S. family members, or applications which rely on priority filing GB 9206318, filed 24 May 1992; see also Vaughn, et al. 1996 , Nature Biotechnology  14: 309-314). Single chain antibodies may also be designed and constructed using available recombinant DNA technology, such as a DNA amplification method (e.g., PCR), or possibly by using a respective hybridoma cDNA as a template. 
     Variant antibodies also are included within the scope of the invention. Thus, variants of the sequences recited in the application also are included within the scope of the invention. Further variants of the antibody sequences having improved affinity can be obtained using methods known in the art and are included within the scope of the invention. For example, amino acid substitutions can be used to obtain antibodies with further improved affinity. Alternatively, codon optimization of the nucleotide sequence can be used to improve the efficiency of translation in expression systems for the production of the antibody. 
     Such variant antibody sequences will share 70% or more (i.e., 80%, 85%, 90%, 95%, 97%, 98%, 99% or greater) sequence identity with the sequences recited in the application. Such sequence identity is calculated with regard to the full length of the reference sequence (i.e., the sequence recited in the application). Percentage identity, as referred to herein, is as determined using BLAST version 2.1.3 using the default parameters specified by the NCBI (the National Center for Biotechnology Information) [Blosum 62 matrix; gap open penalty=11 and gap extension penalty=1]. For example, peptide sequences are provided by this invention that comprise at least about 5, 10, 15, 20, 30, 40, 50, 75, 100, 150, or more contiguous peptides of one or more of the sequences disclosed herein as well as all intermediate lengths there between. As used herein, the term “intermediate lengths” is meant to describe any length between the quoted values, such as 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, etc.; 21, 22, 23, etc.; 30, 31, 32, etc.; 50, 51, 52, 53, etc.; 100, 101, 102, 103, etc.; 150, 151, 152, 153, etc. 
     The present invention provides for antibodies, either alone or in combination with other antibodies, such as, but not limited to, VRC01, anti-V3 loop, CD4bs, and CD4i antibodies as well as PG9/PG16-like antibodies, that have broad neutralizing activity in serum. 
     According to another embodiment, the present invention provides methods for the preparation and administration of an HIV antibody composition that is suitable for administration to a human or non-human primate patient having HIV infection, or at risk of HIV infection, in an amount and according to a schedule sufficient to induce a protective immune response against HIV, or reduction of the HIV virus, in a human. 
     According to another embodiment, the present invention provides a vaccine comprising at least one antibody of the invention and a pharmaceutically acceptable carrier. According to one embodiment, the vaccine is a vaccine comprising at least one antibody described herein and a pharmaceutically acceptable carrier. The vaccine can include a plurality of the antibodies having the characteristics described herein in any combination and can further include antibodies neutralizing to HIV as are known in the art. 
     It is to be understood that compositions can be a single or a combination of antibodies disclosed herein, which can be the same or different, in order to prophylactically or therapeutically treat the progression of various subtypes of HIV infection after vaccination. Such combinations can be selected according to the desired immunity. When an antibody is administered to an animal or a human, it can be combined with one or more pharmaceutically acceptable carriers, excipients or adjuvants as are known to one of ordinary skilled in the art. The composition can further include broadly neutralizing antibodies known in the art, including but not limited to, VRC01, b12, anti-V3 loop, CD4bs, and CD4i antibodies as well as PG9/PG16-like antibodies. 
     Further, with respect to determining the effective level in a patient for treatment of HIV, in particular, suitable animal models are available and have been widely implemented for evaluating the in vivo efficacy against HIV of various gene therapy protocols (Sarver et al. (1993b), supra). These models include mice, monkeys and cats. Even though these animals are not naturally susceptible to HIV disease, chimeric mice models (for example, SCID, bg/nu/xid, NOD/SCID, SCID-hu, immunocompetent SCID-hu, bone marrow-ablated BALB/c) reconstituted with human peripheral blood mononuclear cells (PBMCs), lymph nodes, fetal liver/thymus or other tissues can be infected with lentiviral vector or HIV, and employed as models for HIV pathogenesis. Similarly, the simian immune deficiency virus (SIV)/monkey model can be employed, as can the feline immune deficiency virus (FIV)/cat model. The pharmaceutical composition can contain other pharmaceuticals, in conjunction with a vector according to the invention, when used to therapeutically treat AIDS. These other pharmaceuticals can be used in their traditional fashion (i.e., as agents to treat HIV infection). 
     According to another embodiment, the present invention provides an antibody-based pharmaceutical composition comprising an effective amount of an isolated HIV antibody, or an affinity matured version, which provides a prophylactic or therapeutic treatment choice to reduce infection of the HIV virus. The antibody-based pharmaceutical composition of the present invention may be formulated by any number of strategies known in the art (e.g., see McGoff and Scher, 2000, Solution Formulation of Proteins/Peptides: In McNally, E. J., ed. Protein Formulation and Delivery. New York, N.Y.: Marcel Dekker; pp. 139-158; Akers and Defilippis, 2000, Peptides and Proteins as Parenteral Solutions. In: Pharmaceutical Formulation Development of Peptides and Proteins. Philadelphia, Pa.: Talyor and Francis; pp. 145-177; Akers, et al., 2002, Pharm. Biotechnol. 14:47-127). A pharmaceutically acceptable composition suitable for patient administration will contain an effective amount of the antibody in a formulation which both retains biological activity while also promoting maximal stability during storage within an acceptable temperature range. The pharmaceutical compositions can also include, depending on the formulation desired, pharmaceutically acceptable diluents, pharmaceutically acceptable carriers and/or pharmaceutically acceptable excipients, or any such vehicle commonly used to formulate pharmaceutical compositions for animal or human administration. The diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, physiological phosphate-buffered saline, Ringer&#39;s solutions, dextrose solution, and Hank&#39;s solution. The amount of an excipient that is useful in the pharmaceutical composition or formulation of this invention is an amount that serves to uniformly distribute the antibody throughout the composition so that it can be uniformly dispersed when it is to be delivered to a subject in need thereof. It may serve to dilute the antibody to a concentration which provides the desired beneficial palliative or curative results while at the same time minimizing any adverse side effects that might occur from too high a concentration. It may also have a preservative effect. Thus, for the antibody having a high physiological activity, more of the excipient will be employed. On the other hand, for any active ingredient(s) that exhibit a lower physiological activity, a lesser quantity of the excipient will be employed. 
     The above described antibodies and antibody compositions or vaccine compositions, comprising at least one or a combination of the antibodies described herein, can be administered for the prophylactic and therapeutic treatment of HIV viral infection. 
     The present invention also relates to isolated polypeptides comprising the novel amino acid sequences of the light chains and heavy chains, as well as the consensus sequences for the heavy and light chains of SEQ ID NOs: 1 and 2, as listed in  FIG.  3   . 
     In other related embodiments, the invention provides polypeptide variants that encode the amino acid sequences of the HIV antibodies listed in  FIG.  3   ; the consensus sequences for the heavy and light chains of SEQ ID NOs: 1 and 2. These polypeptide variants have at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%, or greater, sequence identity compared to a polypeptide sequence of this invention, as determined using the methods described herein, (for example, BLAST analysis using standard parameters). One skilled in this art will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by taking into amino acid similarity and the like. 
     The term “polypeptide” is used in its conventional meaning, i.e., as a sequence of amino acids. The polypeptides are not limited to a specific length of the product. Peptides, oligopeptides, and proteins are included within the definition of polypeptide, and such terms can be used interchangeably herein unless specifically indicated otherwise. This term also includes post-expression modifications of the polypeptide, for example, glycosylations, acetylations, phosphorylations and the like, as well as other modifications known in the art, both naturally occurring and non-naturally occurring. A polypeptide can be an entire protein, or a subsequence thereof. Particular polypeptides of interest in the context of this invention are amino acid subsequences comprising CDRs, VH and VL, being capable of binding an antigen or HIV-infected cell. 
     A polypeptide “variant,” as the term is used herein, is a polypeptide that typically differs from a polypeptide specifically disclosed herein in one or more substitutions, deletions, additions and/or insertions. Such variants can be naturally occurring or can be synthetically generated, for example, by modifying one or more of the above polypeptide sequences of the invention and evaluating one or more biological activities of the polypeptide as described herein and/or using any of a number of techniques well known in the art. 
     For example, certain amino acids can be substituted for other amino acids in a protein structure without appreciable loss of its ability to bind other polypeptides (for example, antigens) or cells. Since it is the binding capacity and nature of a protein that defines that protein&#39;s biological functional activity, certain amino acid sequence substitutions can be made in a protein sequence, and, accordingly, its underlying DNA coding sequence, whereby a protein with like properties is obtained. It is thus contemplated that various changes can be made in the peptide sequences of the disclosed compositions, or corresponding DNA sequences that encode said peptides without appreciable loss of their biological utility or activity. 
     In many instances, a polypeptide variant will contain one or more conservative substitutions. A “conservative substitution” is one in which an amino acid is substituted for another amino acid that has similar properties, such that one skilled in the art of peptide chemistry would expect the secondary structure and hydropathic nature of the polypeptide to be substantially unchanged. 
     Amino acid substitutions generally are based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, hydrophilicity, charge, size, and the like. Exemplary substitutions that take various of the foregoing characteristics into consideration are well known to those of skill in the art and include: arginine and lysine; glutamate and aspartate; serine and threonine; glutamine and asparagine; and valine, leucine and isoleucine. 
     “Homology” or “sequence identity” refers to the percentage of residues in the polynucleotide or polypeptide sequence variant that are identical to the non-variant sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology. In particular embodiments, polynucleotide and polypeptide variants have at least about 70%, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least about 98%, or at least about 99% polynucleotide or polypeptide homology with a polynucleotide or polypeptide described herein. 
     Such variant polypeptide sequences will share 70% or more (i.e. 80%, 85%, 90%, 95%, 97%, 98%, 99% or more) sequence identity with the sequences recited in the application. In additional embodiments, the described invention provides polypeptide fragments comprising various lengths of contiguous stretches of amino acid sequences disclosed herein. For example, peptide sequences are provided by this invention that comprise at least about 5, 10, 15, 20, 30, 40, 50, 75, 100, 150, or more contiguous peptides of one or more of the sequences disclosed herein as well as all intermediate lengths there between. 
     The invention also includes nucleic acid sequences encoding part or all of the light and heavy chains of the described inventive antibodies, and fragments thereof. Due to redundancy of the genetic code, variants of these sequences will exist that encode the same amino acid sequences. 
     The present invention also includes isolated nucleic acid sequences encoding the polypeptides for the heavy and light chains of the HIV antibodies listed in  FIG.  3    and the consensus sequences for the heavy and light chains of SEQ ID NOs: 1 and 2. 
     In other related embodiments, the described invention provides polynucleotide variants that encode the peptide sequences of the heavy and light chains of the HIV antibodies listed in  FIG.  3   ; the consensus sequences for the heavy and light chains of SEQ ID NOs: 1 and 2. These polynucleotide variants have at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or greater, sequence identity compared to a polynucleotide sequence of this invention, as determined using the methods described herein, (for example, BLAST analysis using standard parameters). One skilled in this art will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by two nucleotide sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning, and the like. 
     The terms “nucleic acid” and “polynucleotide” are used interchangeably herein to refer to single-stranded or double-stranded RNA, DNA, or mixed polymers. Polynucleotides can include genomic sequences, extra-genomic and plasmid sequences, and smaller engineered gene segments that express, or can be adapted to express polypeptides. 
     An “isolated nucleic acid” is a nucleic acid that is substantially separated from other genome DNA sequences as well as proteins or complexes such as ribosomes and polymerases, which naturally accompany a native sequence. The term encompasses a nucleic acid sequence that has been removed from its naturally occurring environment, and includes recombinant or cloned DNA isolates and chemically synthesized analogues or analogues biologically synthesized by heterologous systems. A substantially pure nucleic acid includes isolated forms of the nucleic acid. Accordingly, this refers to the nucleic acid as originally isolated and does not exclude genes or sequences later added to the isolated nucleic acid by the hand of man. 
     A polynucleotide “variant,” as the term is used herein, is a polynucleotide that typically differs from a polynucleotide specifically disclosed herein in one or more substitutions, deletions, additions and/or insertions. Such variants can be naturally occurring or can be synthetically generated, for example, by modifying one or more of the polynucleotide sequences of the invention and evaluating one or more biological activities of the encoded polypeptide as described herein and/or using any of a number of techniques well known in the art. 
     Modifications can be made in the structure of the polynucleotides of the described invention and still obtain a functional molecule that encodes a variant or derivative polypeptide with desirable characteristics. When it is desired to alter the amino acid sequence of a polypeptide to create an equivalent, or even an improved, variant or portion of a polypeptide of the invention, one skilled in the art typically will change one or more of the codons of the encoding DNA sequence. 
     Typically, polynucleotide variants contain one or more substitutions, additions, deletions and/or insertions, such that the immunogenic binding properties of the polypeptide encoded by the variant polynucleotide is not substantially diminished relative to a polypeptide encoded by a polynucleotide sequence specifically set forth herein. 
     In additional embodiments, the described invention provides polynucleotide fragments comprising various lengths of contiguous stretches of sequence identical to or complementary to one or more of the sequences disclosed herein. For example, polynucleotides are provided by this invention that comprise at least about 10, 15, 20, 30, 40, 50, 75, 100, 150, 200, 300, 400, 500 or 1000 or more contiguous nucleotides of one or more of the sequences disclosed herein as well as all intermediate lengths there between and encompass any length between the quoted values, such as 16, 17, 18, 19, etc.; 21, 22, 23, etc.; 30, 31, 32, etc.; 50, 51, 52, 53, etc.; 100, 101, 102, 103, etc.; 150, 151, 152, 153, etc.; and including all integers through 200-500; 500-1,000. 
     In another embodiment of the invention, polynucleotide compositions are provided that are capable of hybridizing under moderate to high stringency conditions to a polynucleotide sequence provided herein, or a fragment thereof, or a complementary sequence thereof. Hybridization techniques are well known in the art of molecular biology. For purposes of illustration, suitable moderate stringent conditions for testing the hybridization of a polynucleotide of this invention with other polynucleotides include prewashing in a solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridizing at 50-60° C., 5×SSC, overnight; followed by washing twice at 65° C. for 20 minutes with each of 2×, 0.5×, and 0.2×SSC containing 0.1% SDS. One skilled in the art will understand that the stringency of hybridization can be readily manipulated, such as by altering the salt content of the hybridization solution and/or the temperature at which the hybridization is performed. For example, in another embodiment, suitable highly stringent hybridization conditions include those described above, with the exception that the temperature of hybridization is increased, for example, to 60-65° C. or 65-70° C. 
     In some embodiments, the polypeptide encoded by the polynucleotide variant or fragment has the same binding specificity (i.e., specifically or preferentially binds to the same epitope or HIV strain) as the polypeptide encoded by the native polynucleotide. In some embodiments, the described polynucleotides, polynucleotide variants, fragments and hybridizing sequences, encode polypeptides that have a level of binding activity of at least about 50%, at least about 70%, and at least about 90% of that for a polypeptide sequence specifically set forth herein. 
     The polynucleotides of the described invention, or fragments thereof, regardless of the length of the coding sequence itself, can be combined with other DNA sequences, such as promoters, polyadenylation signals, additional restriction enzyme sites, multiple cloning sites, other coding segments, and the like, such that their overall length can vary considerably. A nucleic acid fragment of almost any length is employed. For example, illustrative polynucleotide segments with total lengths of about 10000, about 5000, about 3000, about 2000, about 1000, about 500, about 200, about 100, about 50 base pairs in length, and the like, (including all intermediate lengths) are included in many implementations of this invention. 
     Further included within the scope of the invention are vectors such as expression vectors, comprising a nucleic acid sequence according to the invention. Cells transformed with such vectors also are included within the scope of the invention. 
     The present invention also provides vectors and host cells comprising a nucleic acid of the invention, as well as recombinant techniques for the production of a polypeptide of the invention. Vectors of the invention include those capable of replication in any type of cell or organism, including, for example, plasmids, phage, cosmids, and mini chromosomes. In some embodiments, vectors comprising a polynucleotide of the described invention are vectors suitable for propagation or replication of the polynucleotide, or vectors suitable for expressing a polypeptide of the described invention. Such vectors are known in the art and commercially available. 
     “Vector” includes shuttle and expression vectors. Typically, the plasmid construct also will include an origin of replication (for example, the ColE1 origin of replication) and a selectable marker (for example, ampicillin or tetracycline resistance), for replication and selection, respectively, of the plasmids in bacteria. An “expression vector” refers to a vector that contains the necessary control sequences or regulatory elements for expression of the antibodies including antibody fragment of the invention, in bacterial or eukaryotic cells. 
     As used herein, the term “cell” can be any cell, including, but not limited to, that of a eukaryotic, multicellular species (for example, as opposed to a unicellular yeast cell), such as, but not limited to, a mammalian cell or a human cell. A cell can be present as a single entity, or can be part of a larger collection of cells. Such a “larger collection of cells” can comprise, for example, a cell culture (either mixed or pure), a tissue (for example, endothelial, epithelial, mucosa or other tissue), an organ (for example, lung, liver, muscle and other organs), an organ system (for example, circulatory system, respiratory system, gastrointestinal system, urinary system, nervous system, integumentary system or other organ system), or an organism (e.g., a bird, mammal, or the like). 
     Polynucleotides of the invention may synthesized, whole or in parts that then are combined, and inserted into a vector using routine molecular and cell biology techniques, including, for example, subcloning the polynucleotide into a linearized vector using appropriate restriction sites and restriction enzymes. Polynucleotides of the described invention are amplified by polymerase chain reaction using oligonucleotide primers complementary to each strand of the polynucleotide. These primers also include restriction enzyme cleavage sites to facilitate subcloning into a vector. The replicable vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, and one or more marker or selectable genes. 
     In order to express a polypeptide of the invention, the nucleotide sequences encoding the polypeptide, or functional equivalents, may be inserted into an appropriate expression vector, i.e., a vector that contains the necessary elements for the transcription and translation of the inserted coding sequence. Methods well known to those skilled in the art may be used to construct expression vectors containing sequences encoding a polypeptide of interest and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Such techniques are described, for example, in Sambrook, J., et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, Plainview, N.Y., and Ausubel, F. M. et al. (1989) Current Protocols in Molecular Biology, John Wiley &amp; Sons, New York. N.Y. 
     The present invention also provides kits useful in performing diagnostic and prognostic assays using the antibodies, polypeptides and nucleic acids of the present invention. Kits of the present invention include a suitable container comprising an HIV antibody, a polypeptide or a nucleic acid of the invention in either labeled or unlabeled form. In addition, when the antibody, polypeptide or nucleic acid is supplied in a labeled form suitable for an indirect binding assay, the kit further includes reagents for performing the appropriate indirect assay. For example, the kit may include one or more suitable containers including enzyme substrates or derivatizing agents, depending on the nature of the label. Control samples and/or instructions may also be included. The present invention also provides kits for detecting the presence of the HIV antibodies or the nucleotide sequence of the HIV antibody of the present invention in a biological sample by PCR or mass spectrometry. 
     “Label” as used herein refers to a detectable compound or composition that is conjugated directly or indirectly to the antibody so as to generate a “labeled” antibody. A label can also be conjugated to a polypeptide and/or a nucleic acid sequence disclosed herein. The label can be detectable by itself (for example, radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, can catalyze chemical alteration of a substrate compound or composition that is detectable. Antibodies and polypeptides of the described invention also can be modified to include an epitope tag or label, for example, for use in purification or diagnostic applications. Suitable detection means include the use of labels such as, but not limited to, radionucleotides, enzymes, coenzymes, fluorescers, chemiluminescers, chromogens, enzyme substrates or co-factors, enzyme inhibitors, prosthetic group complexes, free radicals, particles, dyes, and the like. 
     According to another embodiment, the present invention provides diagnostic methods. Diagnostic methods generally involve contacting a biological sample obtained from a patient, such as, for example, blood, serum, saliva, urine, sputum, a cell swab sample, or a tissue biopsy, with an HIV antibody and determining whether the antibody preferentially binds to the sample as compared to a control sample or predetermined cut-off value, thereby indicating the presence of the HIV virus. 
     According to another embodiment, the present invention provides methods to detect the presence of the HIV antibodies of the present invention in a biological sample from a patient. Detection methods generally involve obtaining a biological sample from a patient, such as, for example, blood, serum, saliva, urine, sputum, a cell swab sample, or a tissue biopsy and isolating HIV antibodies or fragments thereof, or the nucleic acids that encode an HIV antibody, and assaying for the presence of an HIV antibody in the biological sample. Also, the present invention provides methods to detect the nucleotide sequence of an HIV antibody in a cell. The nucleotide sequence of an HIV antibody may also be detected using the primers disclosed herein. The presence of the HIV antibody in a biological sample from a patient may be determined utilizing known recombinant techniques and/or the use of a mass spectrometer. 
     In another embodiment, the present invention provides a method for detecting an HIV antibody comprising a heavy chain comprising a highly conserved consensus sequence and a light chain comprising a highly conserved consensus sequence in a biological sample, comprising obtaining an immunoglobulin-containing biological sample from a mammalian subject, isolating an HIV antibody from said sample, and identifying the highly conserved consensus sequences of the heavy chain and the light chain. The biological sample may be blood, serum, saliva, urine, sputum, a cell swab sample, or a tissue biopsy. The amino acid sequences may be determined by methods known in the art including, for example, PCR and mass spectrometry. 
     The term “assessing” includes any form of measurement, and includes determining if an element is present or not. The terms “determining”, “measuring”, “evaluating”, “assessing” and “assaying” are used interchangeably and include quantitative and qualitative determinations. Assessing may be relative or absolute. “Assessing the presence of” includes determining the amount of something present, and/or determining whether it is present or absent. As used herein, the terms “determining,” “measuring,” and “assessing,” and “assaying” are used interchangeably and include both quantitative and qualitative determinations. 
     II. Method of Reducing Viral Replication 
     Methods for reducing an increase in HIV virus titer, virus replication, virus proliferation or an amount of an HIV viral protein in a subject are further provided. According to another aspect, a method includes administering to the subject an amount of an HIV antibody effective to reduce an increase in HIV titer, virus replication or an amount of an HIV protein of one or more HIV strains or isolates in the subject. 
     According to another embodiment, the present invention provides a method of reducing viral replication or spread of HIV infection to additional host cells or tissues comprising contacting a mammalian cell with the antibody, or a portion thereof, which binds to an antigenic epitope on gp120. 
     III. Method of Treatment 
     According to another embodiment, the present invention provides a method for treating a mammal infected with a virus infection, such as, for example, HIV, comprising administering to said mammal a pharmaceutical composition comprising the HIV antibodies disclosed herein. According to one embodiment, the method for treating a mammal infected with HIV comprises administering to said mammal a pharmaceutical composition that comprises an antibody of the present invention, or a fragment thereof. The compositions of the invention can include more than one antibody having the characteristics disclosed (for example, a plurality or pool of antibodies). It also can include other HIV neutralizing antibodies as are known in the art, for example, but not limited to, VRC01, PG9 and b12. 
     Passive immunization has proven to be an effective and safe strategy for the prevention and treatment of viral diseases. (See, for example, Keller et al., Clin. Microbiol. Rev. 13:602-14 (2000); Casadevall, Nat. Biotechnol. 20:114 (2002); Shibata et al., Nat. Med. 5:204-10 (1999); and Igarashi et al., Nat. Med. 5:211-16 (1999). Passive immunization using human monoclonal antibodies provides an immediate treatment strategy for emergency prophylaxis and treatment of HIV. 
     Subjects at risk for HIV-related diseases or disorders include patients who have come into contact with an infected person or who have been exposed to HIV in some other way. Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic of HIV-related disease or disorder, such that a disease or disorder is prevented or, alternatively, delayed in its progression. 
     For in vivo treatment of human and non-human patients, the patient is administered or provided a pharmaceutical formulation including an HIV antibody of the invention. When used for in vivo therapy, the antibodies of the invention are administered to the patient in therapeutically effective amounts (i.e., amounts that eliminate or reduce the patient&#39;s viral burden). The antibodies are administered to a human patient, in accord with known methods, such as intravenous administration, for example, as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intracerobrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes. The antibodies can be administered parenterally, when possible, at the target cell site, or intravenously. In some embodiments, antibody is administered by intravenous or subcutaneous administration. Therapeutic compositions of the invention may be administered to a patient or subject systemically, parenterally, or locally. The above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician. 
     For parenteral administration, the antibodies may be formulated in a unit dosage injectable form (solution, suspension, emulsion) in association with a pharmaceutically acceptable, parenteral vehicle. Examples of such vehicles include, but are not limited, water, saline, Ringer&#39;s solution, dextrose solution, and 5% human serum albumin. Nonaqueous vehicles include, but are not limited to, fixed oils and ethyl oleate. Liposomes can be used as carriers. The vehicle may contain minor amounts of additives such as substances that enhance isotonicity and chemical stability, such as, for example, buffers and preservatives. The antibodies can be formulated in such vehicles at concentrations of about 1 mg/ml to 10 mg/ml. 
     The dose and dosage regimen depends upon a variety of factors readily determined by a physician, such as the nature of the infection, for example, its therapeutic index, the patient, and the patient&#39;s history. Generally, a therapeutically effective amount of an antibody is administered to a patient. In some embodiments, the amount of antibody administered is in the range of about 0.1 mg/kg to about 50 mg/kg of patient body weight. Depending on the type and severity of the infection, about 0.1 mg/kg to about 50 mg/kg body weight (for example, about 0.1-15 mg/kg/dose) of antibody is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. The progress of this therapy is readily monitored by conventional methods and assays and based on criteria known to the physician or other persons of skill in the art. The above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician. 
     Other therapeutic regimens may be combined with the administration of the HIV antibody of the present invention. The combined administration includes co-administration, using separate formulations or a single pharmaceutical formulation, and consecutive administration in either order, wherein preferably there is a time period while both (or all) active agents simultaneously exert their biological activities. Such combined therapy can result in a synergistic therapeutic effect. The above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician. 
     The terms “treating” or “treatment” or “alleviation” are used interchangeably and refer to both therapeutic treatment and prophylactic or preventative measures; wherein the object is to prevent or slow down (lessen) the targeted pathologic condition or disorder. Those in need of treatment include those already with the disorder as well as those prone to have the disorder or those in whom the disorder is to be prevented. A subject or mammal is successfully “treated” for an infection if, after receiving a therapeutic amount of an antibody according to the methods of the present invention, the patient shows observable and/or measurable reduction in or absence of one or more of the following: reduction in the number of infected cells or absence of the infected cells; reduction in the percent of total cells that are infected; and/or relief to some extent, one or more of the symptoms associated with the specific infection; reduced morbidity and mortality, and improvement in quality of life issues. The above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician. 
     The term “therapeutically effective amount” refers to an amount of an antibody or a drug effective to treat a disease or disorder in a subject or mammal. 
     Administration “in combination with” one or more further therapeutic agents includes simultaneous (concurrent) and consecutive administration in any order. 
     “Carriers” as used herein include pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include, but not limited to, buffers such as phosphate, citrate, and other organic acids; antioxidants including, but not limited to, ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as, but not limited to, serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as, but not limited to, polyvinylpyrrolidone; amino acids such as, but not limited to, glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including, but not limited to, glucose, mannose, or dextrins; chelating agents such as, but not limited to, EDTA; sugar alcohols such as, but not limited to, mannitol or sorbitol; salt-forming counterions such as, but not limited to, sodium; and/or nonionic surfactants such as, but not limited to, TWEEN.; polyethylene glycol (PEG), and PLURONICS. 
     Where a value of ranges is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges which may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the invention. 
     Example 1 
     This example describes materials and methods used in EXAMPLES 2-5 below. 
     HIV antibodies were cloned and produced following gp140-specific single B-cell capture as previously described (Mouquet, H. et al. PLoS One 6, e24078 (2011); Tiller, T. et al. J Immunol Methods 329, 112-24 (2008); and Scheid, J. F. et al. Nature 458, 636-40 (2009)). PGT121 GM  and 10-1074 GM  “glycomutant” antibodies were generated by substituting 10-1074 residues at HC positions 32, 53, 54, 58, 97, 1001 into PGT121 and vice versa. Binding properties of anti-gp140 antibodies to HIV Env proteins were assayed by ELISA, SPR and glycan microarray assays as previously described (Scheid, J. F. et al. Science 333, 1633-7 (2011); Walker, L. M. et al. Nature 477, 466-70 (2011); and Mouquet, H. et al. PLoS One 6, e24078 (2011)). Neutralization was evaluated using (i) a luciferase-based assay in TZM.b1 cells, and (ii) a PBMC-based assay using infection with primary HIV-1 variants as previously described (Li, M. et al. J Virol 79, 10108-25 (2005); Euler, Z. et al. Journal of virology 85, 7236-45 (2011); and Bunnik, E. M. et al. Nature medicine 16, 995-7 (2010)). Structures of PGT121 (“unliganded” and “liganded”), 10-1074 and GL Fab fragments were solved by molecular replacement to 2.8 Å, 2.3 Å, 1.8 Å and 2.4 Å resolution, respectively. 
     Single B Cell RT-PCRs and Ig Gene Analyses 
     Single-cell sorting of gp140 + CD19 + IgG + B cells from patient 10 (pt10; referred to as patient 17 in Nature 477(7365):466-470.) PBMCs, cDNA synthesis and nested PCR amplifications of Ig genes were performed in a previous study (PLoS One 6(9):e24078). IGλ genes expressed by PGT121 clonal variants were PCR amplified using a forward primer (L-Vλ3-21*02: 5′ CTGGACCGTTCTCCTCCTCG 3′ (SEQ ID NO: 137)) further upstream in the leader region to avoid the potentially mutated region (31). All PCR products were sequenced and analyzed for Ig gene usage, CDR3 analyses and number of VH/Vκ somatic hypermutations (IgBLAST and IMGT®). Multiple sequence alignments were performed using the MacVector program (v.12.5.0) with the ClustalW analysis function (default parameters), and were used to generate dendrograms by the Neighbor Joining method (with Best tree mode and outgroup rooting). Alternatively, dendrograms were generated using the UPGMA method (with Best tree mode). 
     The germline (GL) precursor gene segments of the PGT121-like and 10-1074-like antibodies were identified using IgBLAST and IMGT®/V-QUEST as V H 4-59*01, J H 6*03, V L 3-21*02 and J L 3*02. (These gene segments are among the most frequently used in the repertoire of human antibodies (PLoS One 6(8):e22365; Immunogenetics 64(5):337-350). To build a representative GL ancestor sequence, we aligned the IgH and IgL sequences of 10-996 (the antibody containing the fewest somatic hypermutations) to the GL sequences using IgBLAST. The GL IgH sequence was constructed by replacing the mature V H  and J H  gene segments with their GL counterparts and using the 10-996 sequence for the CDRH3 region involving N-region nucleotides and the DH gene segment. The GL IgL sequence was assembled from the V L 3-21*02 and J L 3*02 gene segment sequences. 
     Cloning and Production of Antibodies 
     Purified digested PCR products were cloned into human Igγ 1 - or Igλ-expressing vectors (J Immunol Methods 329(1-2):112-124). Vectors containing IgH and Igλ genes were then sequenced and compared to the original PCR product sequences. PGT121 and 10-303 shared the same Igλ gene and had one amino acid difference in position 2 of the IgH gene ( FIG.  4   ); therefore to produce the PGT121 IgG, we used the 10-303 Igλ gene and a PGT121 IgH gene generated by introducing a single substitution (V2M) into the 10-303 IgH gene by site-directed mutagenesis (QuikChange Site-Directed Mutagenesis Kit; Stratagene). To generate His-tagged Fabs, the PGT121 and 10-1074 V H  genes were subcloned into a 6×His-IgCγ1 (‘6×His’ disclosed as SEQ ID NO: 138) expression vector generated by modifying our standard Igγ 1  vector (Science 301(5638):1374-1377) to encode the IgG1 C H 1 domain followed by a 6×-His tag (SEQ ID NO: 138). IgH DNA fragments encoding PGT121 GM  (S32Y, K53D, S54R, N58T, H97R, T1001Y) and 10-1074 GM  (Y32S, D53K, R54S, T58N, R97H, Y1001T) mutant antibodies were obtained as a synthetic minigene (IDT) and subcloned into Igγ 1 -expressing vectors. 
     Listed below is the heavy chain sequence for 10-1074 GM  where the mutations are underlined. The light chain sequence of 10-1074 GM  is the same as that of 10-1074. 
     
       
         
           
               
            
               
                 (SEQ ID NO: 129) 
               
               
                 QVQLQESGPGLVKPSETLSVTCSVSGDSMNN S YWTWIRQSPGKGLEWI 
               
               
                   
               
               
                 GYIS KS ESA N YNPSLNSRVVISRDTSKNQLSLKLNSVTPADTAVYYCA 
               
               
                   
               
               
                 TAR H GQRIYGVVSFGEFF T YYSMDVWGKGTTVTVSS 
               
            
           
         
       
     
     Antibodies and Fab fragments were produced by transient transfection of IgH and IgL expression plasmids into exponentially growing HEK 293T cells (ATCC, CRL-11268) using the polyethyleneimine (PEI)-precipitation method (PLoS One 6(9):e24078). IgG antibodies were affinity purified using Protein G sepharose beads (GE Healthcare) according to the manufacturer&#39;s instructions. Fab fragments were affinity purified using HisPur™ Cobalt Resin (Thermo scientific) as described below. 
     HIV-1 Env Proteins 
     Alanine mutations were introduced into the pYU-2 gp120 vector (gift of J. Sodroski, Harvard Medical School) at positions 301 to 303 (Asn-Asn-Thr), 324 to 325 (Gly-Asp), and 332 (Asn) (HXBc2 amino acid numbering) using the QuikChange Site-Directed Mutagenesis kit (Stratagene) according to the manufacturer&#39;s instructions. The same procedure was used to generate “double glycan” mutants by introducing single alanine mutations in the pYU-2 gp120 N332A  vector at each PNGS located between Asn262 gp120  and Asn406 gp120 . Site-directed mutations were verified by DNA sequencing. 
     Expression vectors encoding YU-2 gp140 (Journal of virology 74(12):5716-5725), YU-2 gp120, HXB2c gp120 core  (Nature 393(6686):648-659), HXB2c 2CCcore (PLoS Pathog 5(5):e1000445) proteins, and YU-2 gp120 mutant proteins were used to transfect HEK 293T cells. To produce high-mannose-only YU-2 gp120 protein (gp120 kif ), 25 μM kifunensine (Enzo Life Sciences) was added at the time of transfection. Culture supernatants were harvested and concentrated using centrifugation-based filtration devices (Vivacell 100, Sartorius Stedim Biotech Gmbh) that allowed buffer exchange of the samples into 10 mM imidazole, 50 mM sodium phosphate, 300 mM sodium chloride; pH 7.4. Proteins were purified by affinity chromatography using HisPur™ Cobalt Resin (Thermo scientific) according to the manufacturer&#39;s instructions. 
     For deglycosylation reactions, 50 μg of HEK 293T cell-produced YU-2 gp120 in PBS was digested overnight at 37° C. with 200 U of PNGase F (New England Biolabs) or 10,000 U of Endo H f  (New England Biolabs) in their respective reaction buffers without denaturing agents. After buffer exchange into PBS using Centrifugal Filters (Amicon® Ultra, Millipore), glycosidase-treated gp120s (200 ng) were examined by SDS-PAGE using a 4-12% NuPAGE gel (Invitrogen) followed by silver staining (Pierce Silver Stain Kit, Thermo Scientific). 
     ELISAs 
     High-binding 96-well ELISA plates (Costar) were coated overnight with 100 ng/well of purified gp120 in PBS. After washing, the plates were blocked for 2 h with 2% BSA, 1 μM EDTA, 0.05% Tween-PBS (blocking buffer) and then incubated for 2 h with IgGs at concentrations of 26.7 nM (or 427.2 nM for ELISAs using the YU-2 gp120 double glycan mutants) and 7 consecutive 1:4 dilutions in PBS. After washing, the plates were developed by incubation with goat HRP-conjugated anti-human IgG antibodies (Jackson ImmunoReseach) (at 0.8 μg/ml in blocking buffer) for 1 h, and by addition of HRP chromogenic substrate (ABTS solution, Invitrogen) (PLoS One 6(9):e24078). Antibody binding to the selected gp120 V3  overlapping peptides was tested using a previously described peptide-ELISA method. 
     For competition ELISAs, gp120-coated plates were blocked for 2 h with blocking buffer and then incubated for 2 h with biotinylated antibodies (at a concentration of 26.6 nM for PGT121, 0.21 nM for 10-1074, 0.43 nM for 10-996 and 1.67 nM for 10-1369) in 1:2 serially diluted solutions of antibody competitors in PBS (IgG concentration range from 5.2 to 667 nM). Plates were developed as described above using HRP-conjugated streptavidin (Jackson ImmunoReseach) (at 0.8 μg/ml in blocking buffer). All experiments were performed at least in duplicate. 
     Glycan Microarray Analysis 
     Microarrays were generated by robotically printing glycan probes linked to lipid (neoglycolipids) onto nitrocellulose-coated glass slides (Methods Mol Biol 808:117-136) at two levels (2 and 5 fmol/spot) in duplicate. Binding assays were performed with microarrays containing 15 neoglycolipids derived from N-glycans of high-mannose and complex-types. The sequences of the probes are shown in  FIG.  7 A . In brief, antibodies were tested at 50 μg/ml, and binding was detected with biotinylated anti-human IgG (Vector) followed by AlexaFluor 647-labeled streptavidin (Molecular Probes). 
     Surface Plasmon Resonance 
     Experiments were performed using a Biacore T100 (Biacore, Inc)(Nature 467(7315):591-595). Briefly, YU-2 gp140 and gp120 proteins were primary amine-coupled on CMS chips (Biacore, Inc.) at a coupling density of 300 RUs. Anti-gp120 IgGs and the germline precursor (GL) were injected over flow cells at 1 μM and 10 μM, respectively, at flow rates of 35 μl/min with 3 min association and 5 min dissociation phases. The sensor surface was regenerated by a 30 sec injection of 10 mM glycine-HCl pH 2.5 at a flow rate of 50 μl/min. Dissociation (k d  (s −1 )), association (k a  (M −1  s −1 ) and binding constants (K D  (M) or K A  (M −1 ) were calculated from kinetic analyses after subtraction of backgrounds using a 1:1 binding model without a bulk reflective index (RI) correction (Biacore T100 Evaluation software). Binding constants for bivalent IgGs calculated using a 1:1 binding model are referred to in the text as “apparent” affinities to emphasize that the K D  values include potential avidity effects 
     Neutralization Assays 
     Virus neutralization was evaluated using a luciferase-based assay in TZM.b1 cells (J Virol 79(16):10108-10125). The HIV-1 pseudoviruses tested contained mostly tier-2 and tier-3 viruses (Journal of virology 84(3):1439-1452)(Tables 4 and 5). High-mannose-only pseudoviruses were produced in wild-type cells treated with 25 μM kifunensine (Enzo Life Sciences) ( FIG.  8 C ) or in HEK 293S GnTI −/−  cells ( FIG.  8 D ). Non-linear regression analysis was used to calculate concentrations at which half-maximal inhibition was observed (IC 50  values). Neutralization activities were also evaluated with a previously characterized PBMC-based assay using infection with primary HIV-1 variants (n=95) isolated from clade B-infected donors with known seroconversion dates either between 1985 and 1989 (“historical seroconverters”, n=14) or between 2003 and 2006 (“contemporary seroconverters”, n=21)(Journal of virology 85(14):7236-7245; Nat Med 16(9):995-997). Neutralization activity for each antibody was calculated using GraphPad Prism software (v5.0b) as area under the best-fit curve, which fits the proportion of viruses neutralized over IC 50  values ranging from 0.001 to 50 μg/ml. Relative area under the curve (RAUC) values were derived by normalizing all AUC values by the highest value (obtained with 10-1074). 
     Statistical Analyses 
     Statistical analyses were performed with the GraphPad Prism software (v5.0b). Neutralization potencies in the TZM-bl assay against the selected panel of 9 virus strains versus the apparent binding affinities of the antibodies for gp120 and gp140 were analyzed using a Spearman&#39;s correlation test. The Mann Whitney test was used to compare: (i) affinities for gp120/gp140 of antibodies belonging to the PGT121 or 10-1074 group, and (ii) neutralization activities against viruses isolated from historical and contemporary seroconverters. 
     Crystallization and Structure Determinations 
     6×-His (SEQ ID NO: 138) tagged PGT121, 10-1074 and 10-996GL Fabs for crystallization were expressed. Fabs were purified from the supernatants of transiently-transfected HEK 293-6E cells by sequential Ni 2+ -NTA affinity (Qiagen) and Superdex200 10/300 (GE Healthcare) size exclusion chromatography. For crystals of the unliganded PGT121 Fab, PGT121 IgG was isolated from the supernatants of transiently-transfected HEK 293-6E cells by Protein A affinity chromatography (Pierce), and Fab fragments were obtained by papain cleavage of the IgG and further purification using Superdex200 10/300 (GE Healthcare) size exclusion chromatography. 
     Purified Fabs were concentrated to 8-20 mg/mL (“unliganded” PGT121, 8 mg/mL; 10-1074 and GL, 20 mg/mL) in PBS buffer. The “liganded” PGT121 Fab crystals were prepared from a protein sample (final concentration: 15 mg/mL) that was mixed with a 3-fold molar excess of NA2 glycan and incubated at 20° C. for 2 hours. Crystallization conditions were screened at 20° C. using a Mosquito® crystallization robot (TTP labs) in 400 nL drops using a 1:1 protein to reservoir ratio. Crystals of “unliganded” PGT121 Fab (P2 1 2 1 2 1 ; a=56.8, b=74.7, c=114.9 Å) were obtained in 24% PEG 4,000, 0.1 M Tris-HCl pH 8.5, 10 mM CuCl 2  and crystals of “liganded” PGT121 Fab (P2 1 2 1 2 1 ; a=67.8, b=67.8, c=94.1 Å) grew in 17% PEG 10,000, 0.1M Bis-Tris pH 5.5, 0.1M CH 3 COOHNH 4 . Crystals of 10-1074 Fab (P21; a=61.4, b=40.3, c=84.5 Å; β=95.39°) were obtained in 25% PEG 3,350, 0.1 M Bis-Tris pH 5.5, 0.2 M NaCl, and crystals of GL Fab (P21; a=54.9, b=344.7, c=55.2 Å; β=91.95°) grew in 20% PEG 3,350, 0.24 M sodium malonate pH 7.0, 10 mM MnCl 2 . Crystals were cryoprotected by soaking in mother liquor containing 20% glycerol (“unliganded” and “liganded” PGT121 Fab) or 20% ethylene glycol (10-1074 Fab and GL Fab) and subsequently flash-cooled in liquid nitrogen. 
     Diffraction data were collected at beamline 12-2 (wavelength=1.029 Å) at the Stanford Synchrotron Radiation Lightsource (SSRL) on a Pilatus 6M pixel detector (Dectris). Data were indexed, integrated and scaled using XDS. Using the data obtained from the “unliganded” PGT121 Fab crystals, we used Phenix to find a molecular replacement solution for one Fab per asymmetric unit (chains H and L for the heavy and light chain, respectively) using two search models, the C H -C L  domains of PGT128 Fab (PDB code 3PV3) and the V H -V L  domains of 2F5 (PDB code 3IDJ) after omitting residues in the CDRH3 and CDRL3 loops. Subsequently, we used the “unliganded” PGT121 structure as a search model to find molecular replacement solutions for “liganded” PGT121 Fab (one Fab per asymmetric unit), 10-1074 Fab (one Fab per asymmetric unit) and GL (four Fabs per asymmetric unit). 
     Iterative refinement (including non-crystallographic symmetry restraints for GL) was performed using Phenix and manually fitting models into electron density maps using Coot. The atomic models were refined to 3.0 Å resolution for PGT121 Fab (R work =21.6%; R free =26.4%), 1.9 Å resolution for 10-1074 Fab (R work =18.7%; R free =22.3%), 2.4 Å resolution for four GL Fab molecules (R work =19.4%; R free =23.7%), and 2.4 Å resolution for “liganded” PGT121 Fab (R work =20.1%; R free =24.9%). The atomic model of PGT121 Fab contains 95.2%, 4.9% and 0.0% of the residues in the favored, allowed and disallowed regions of the Ramachandran plot, respectively (10-1074 Fab: 98.8%, 0.9%, 0.2%; GL Fab: 96.0%, 3.8%, 0.23%; “liganded PGT121 Fab: 96.7%, 3.1%, 0.2%). PyMOL was used for molecular visualization and to generate figures of the Fab structures. Buried surface area calculations were performed with Areaimol (CCP4 Suite) using a 1.4 Å probe. 
     Fab structures were aligned using the Super script in PyMOL. Pairwise Ca alignments were performed using PDBeFold. 
     Example 2 Predominance and Diversity of PGT121 Clonotype 
     gp140-specific IgG memory B cells were isolated from a clade A-infected African donor using YU-2 gp140 trimers as “bait.” Eighty-seven matching immunoglobulin heavy (IgH) and light (IgL) chain genes corresponding to 23 unique clonal families were identified. The IgH anti-gp140 repertoire was dominated by one clonal family representing ˜28% of all expanded B cell clones. This B cell family corresponds to the same clone as PGT121-123 (Nature 477(7365):466-470) and contained 38 members, 29 of which were unique variants at the nucleotide level (Table 3). Based on their IgH nucleotide sequence, the PGT121 family divides into two groups: a PGT121-like group containing PGT121-123 and 9 closely-related variants, and a second group, 10-1074-like, containing 20 members. Although our traditional primers (J Immunol Methods 329(1-2):112-124; Science 301(5638):1374-1377) did not amplify the IgL genes expressed by the PGT121 B cell clone due to the nucleotide deletions in the region encoding framework region 1, 24 of 38. genes were obtained using new Igλ-specific primers designed to amplify heavily somatically-mutated genes (Table 3). Consistent with the high levels of hypermutation in the IgH genes (18.2% of the VH gene on average), the amplified Igλ genes were highly mutated (18.2% of the Vλ gene on average) and carried nucleotide deletions in framework region 1 (FWR1) (12 to 21 nucleotides) and a 9-nucleotide insertion in framework region 3 (FWR3) ( FIG.  3 B  and Table 3). 
     The sequence alignments of three PGT antibodies (PGT-121, -122, and -123), eleven PGT121 and 10-1074 clonal variants (10-259, 10-303, 10-410, 10-847, 10-996, 10-1074, 10-1121, 10-1130, 10-1146, 10-1341, 10-1369, and 10-1074 GM ), likely germline (GL), and consensus sequences are shown in  FIGS.  3 ( a ) and  3 ( b ) . The sequences for corresponding heavy chain variable regions, light chain variable regions, heavy chain CDRs, and light chain CDRs under both IMGT and KABT systems are listed in Table 1 below. Assigned sequence identification numbers for the sequences under the KABT systems are listed in Table 2 below: 
     
       
         
           
               
             
               
                 TABLE 1  
               
               
                   
               
             
            
               
                 IgH SEQUENCES 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 IMGT 
                 FWR1 
                 CDR1 
                 FWR2 
                 CDR2 
                 FWR3 
                 CDR3 
                 FWR4 
               
               
                   
               
               
                 10-1369 
                 QVQLQESGPGLVKPLETLSLTCN 
                 GAFIADHY 
                 WSWIRLPLGKG 
                 VHDSGDI 
                 NYNPSLKNRVHLSLDKSTNQVSLKLM 
                 ATTKHGRRIYGVVAFGE 
                 WGRGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 PEWIGY 
                 (SEQ ID 
                 AVTAGDSALYYC 
                 WFTYFYMDV (SEQ ID 
                   
               
               
                 NO: 23) 
                   
                 139) 
                   
                 NO: 140) 
                   
                 NO: 141) 
                   
               
               
                   
               
               
                 10-259 
                 QVHLQESGPGLVKPSETLSLTCN 
                 GTLVRDNY 
                 WSWMRQPLGKQ 
                 VHDSGDT 
                 NYNPSLKSRVHLSLDKSNNLVSLRLT 
                 ATTKHGRRIYGIVAFNE 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 PEWIGY 
                 (SEQ ID 
                 AVTAADSATYYC 
                 WFTYFYMDV (SEQ ID 
                   
               
               
                 NO: 3) 
                   
                 142) 
                   
                 NO: 143) 
                   
                 NO: 144) 
                   
               
               
                   
               
               
                 10-303 
                 QVQLQESGPGLVKPSETLSLTCS 
                 GASISDSY 
                 WSWIRRSPGKG 
                 VHKSGDT 
                 NYSPSLKSRVNLSLDTSKNQVSLSLV 
                 ARTLHGRRIYGIVAFNE 
                 WGNGTQVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 AATAADSGKYYC 
                 WFTYFYMDV (SEQ ID 
                   
               
               
                 NO: 5) 
                   
                 145) 
                   
                 NO: 146) 
                   
                 NO: 147) 
                   
               
               
                   
               
               
                 10-410 
                 QVQLQESGPGLVKPPETLSLTCS 
                 GASVNDAY 
                 WSWIRQSPGKR 
                 VHHSGDT 
                 NYNPSLKRRVTFSLDTAKNEVSLKLV 
                 ARALHGKRIYGIVALGE 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 PEWVGY 
                 (SEQ ID 
                 ALTAADSAVYFC 
                 LFTYFYMDV (SEQ ID 
                   
               
               
                 NO: 7) 
                   
                 148) 
                   
                 NO: 149) 
                   
                 NO: 150) 
                   
               
               
                   
               
               
                 10-1130 
                 QVQLQESGPGLVKPPETLSLTCS 
                 GASINDAY 
                 WSWIRQSPGKR 
                 VHHSGDT 
                 NYNPSLKRRVTFSLDTAKNEVSLKLV 
                 ARALHGKRIYGIVALGE 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 PEWVGY 
                 (SEQ ID 
                 DLTAADSAVYFC 
                 LFTYFYMDV (SEQ ID 
                   
               
               
                 NO: 17) 
                   
                 151) 
                   
                 NO: 152) 
                   
                 NO: 153) 
                   
               
               
                   
               
               
                 10-1121 
                 QVQLQESGPGLVKPPETLSLTCS 
                 GASINDAY 
                 WSWIRQSPGKR 
                 VHHSGDT 
                 NYNPSLKRRVSFSLDTAKNEVSLKLV 
                 ARALHGKRIYGIVALGE 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 PEWVGY 
                 (SEQ ID 
                 DLTAADSAIYFC 
                 LFTYFYMDV (SEQ ID 
                   
               
               
                 NO: 15) 
                   
                 154) 
                   
                 NO: 155) 
                   
                 NO: 156) 
                   
               
               
                   
               
               
                 10-1146 
                 QVQLVESGPGLVTPSETLSLTCT 
                 NGSVSGRF 
                 WSWIRQSPGRG 
                 FSDTDRS 
                 EYSPSLRSRLTLSLDASRNQLSLKLK 
                 ARAQQGKRIYGIVSFGE 
                 WGKGTAVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 SVTAADSATYYC 
                 FFYYYYMDA (SEQ ID 
                   
               
               
                 NO: 19) 
                   
                 157) 
                   
                 NO: 158) 
                   
                 NO: 159) 
                   
               
               
                   
               
               
                 10-996 
                 QVQLQESGPGLVKPSETLSLTCS 
                 NGSVSGRF 
                 WSWIRQSPGRG 
                 FSDTEKS 
                 NYNPSLRSRLTLSVDASKNQLSLKLN 
                 ARTQQGKRIYGVVSFGE 
                 WGKGTAVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 SVTAADSATYYC 
                 FFHYYYMDA (SEQ ID 
                   
               
               
                 NO: 11) 
                   
                 160) 
                   
                 NO: 161) 
                   
                 NO: 162) 
                   
               
               
                   
               
               
                 GL (SEQ 
                 QVQLQESGPGLVKPSETLSLTCT 
                 GGSISSYY 
                 WSWIRQPPGKG 
                 IYYSGST 
                 NYNPSLKSRVTISVDTSKNQFSLKLS 
                 ARTQQGKRIYGVVSFGD 
                 WGKGTTVTVSS 
               
               
                 ID NO: 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 SVTAADTAVYYC 
                 YYYYYYMDV (SEQ ID 
                   
               
               
                 31) 
                   
                 163) 
                   
                 NO: 164) 
                   
                 NO: 165) 
                   
               
               
                   
               
               
                 10-1341 
                 QVQLQESGPGLVKPSETLSVTCS 
                 GDSMNNYY 
                 WTWIRQSPGKG 
                 ISDRESA 
                 TYNPSLNSRVVISRDTSTNQLSLKLN 
                 ATARRGQRIYGVVSFGE 
                 WGRGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 SVTPADTAVYYC 
                 FFYYYSMDV (SEQ ID 
                   
               
               
                 NO: 21) 
                   
                 166) 
                   
                 NO: 167) 
                   
                 NO: 168) 
                   
               
               
                   
               
               
                 10-847 
                 QVQLQESGPGLVKPSETLSVTCS 
                 GDSMNNYY 
                 WTWIRQSPGKG 
                 ISDRASA 
                 TYNPSLNSRVVISRDTSKNQLSLKLN 
                 ATARRGQRIYGVVSFGE 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 SVTPADTAVYYC 
                 FFYYYSMDV (SEQ ID 
                   
               
               
                 NO: 9) 
                   
                 169) 
                   
                 NO: 170) 
                   
                 NO: 171) 
                   
               
               
                   
               
               
                 10-1074 
                 QVQLQESGPGLVKPSETLSVTCS 
                 GDSMNNYY 
                 WTWIRQSPGKG 
                 ISDRESA 
                 TYNPSLNSRVVISRDTSKNQLSLKLN 
                 ATARRGQRIYGVVSFGE 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 SVTPADTAVYYC 
                 FFYYYSMDV (SEQ ID 
                   
               
               
                 NO: 13) 
                   
                 172) 
                   
                 NO: 173) 
                   
                 NO: 174) 
                   
               
               
                   
               
               
                 10- 
                 QVQLQESGPGLVKPSETLSVTCS 
                 GDSMNNSY 
                 WTWIRQSPGKG 
                 ISKSESA 
                 NYNPSLNSRVVISRDTSKNQLSLKLN 
                 ATARHGQRIYGVVSFGE 
                 WGKGTTVTVSS 
               
               
                 1074GM 
                 VS 
                 (SEQ ID NO: 
                 LEWIGY 
                 (SEQ ID 
                 SVTPADTAVYYC 
                 FFTYYSMDV (SEQ ID 
                   
               
               
                 (SEQ ID 
                   
                 175) 
                   
                 NO: 176) 
                   
                 NO: 177) 
                   
               
               
                 NO: 129) 
               
               
                   
               
               
                 KABAT 
                 FWR1 
                 CDR1 
                 FWR2 
                 CDR2 
                 FWR3 
                 CDR3 
                 FWR4 
               
               
                   
               
               
                 10-1369 
                 QVQLQESGPGLVKPLETLSLTCN 
                 DHYWS (SEQ 
                 WIRLPLGKGPE 
                 YVHDSGDINY 
                 RVHLSLDKSTNQVSLKLMAVTAGDSA 
                 TKHGRRIYGVVAFGEWF 
                 WGRGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGAFIA 
                 ID NO: 99) 
                 WIG 
                 NPSLKN 
                 LYYCAT 
                 TYFYMDV (SEQ ID 
                   
               
               
                 NO: 23) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 101) 
                   
               
               
                   
                   
                   
                   
                 NO: 100) 
                   
                   
                   
               
               
                   
               
               
                 10-259 
                 QVHLQESGPGLVKPSETLSLTCN 
                 DNYWS (SEQ 
                 WMRQPLGKQPE 
                 YVHDSGDTNY 
                 RVHLSLDKSNNLVSLRLTAVTAADSA 
                 TKHGRRIYGIVAFNEWF 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGTLVR 
                 ID NO: 39) 
                 WIG 
                 NPSLKS 
                 TYYCAT 
                 TYFYMDV (SEQ ID 
                   
               
               
                 NO: 3) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 41) 
                   
               
               
                   
                   
                   
                   
                 NO: 40) 
                   
                   
                   
               
               
                   
               
               
                 10-303 
                 QVQLQESGPGLVKPSETLSLTCS 
                 DSYWS (SEQ 
                 WIRRSPGKGLE 
                 YVHKSGDTNY 
                 RVNLSLDTSKNQVSLSLVAATAADSG 
                 TLHGRRIYGIVAFNEWF 
                 WGNGTQVTVSS 
               
               
                 (SEQ ID 
                 VSGASIS 
                 ID NO: 45) 
                 WIG 
                 SPSLKS 
                 KYYCAR 
                 TYFYMDV (SEQ ID 
                   
               
               
                 NO: 5) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 47) 
                   
               
               
                   
                   
                   
                   
                 NO: 46) 
                   
                   
                   
               
               
                   
               
               
                 10-410 
                 QVQLQESGPGLVKPPETLSLTCS 
                 DAYWS (SEQ 
                 WIRQSPGKRPE 
                 YVHHSGDTNY 
                 RVTFSLDTAKNEVSLKLVALTAADSA 
                 ALHGKRIYGIVALGELF 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGASVN 
                 ID NO: 51) 
                 WVG 
                 NPSLKR 
                 VYFCAR 
                 TYFYMDV (SEQ ID 
                   
               
               
                 NO: 7) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 53) 
                   
               
               
                   
                   
                   
                   
                 NO: 52) 
                   
                   
                   
               
               
                   
               
               
                 10-1130 
                 QVQLQESGPGLVKPPETLSLTCS 
                 DAYWS (SEQ 
                 WIRQSPGKRPE 
                 YVHHSGDTNY 
                 RVTFSLDTAKNEVSLKLVDLTAADSA 
                 ALHGKRIYGIVALGELF 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGASIN 
                 ID NO: 81) 
                 WVG 
                 NPSLKR 
                 VYFCAR 
                 TYFYMDV (SEQ ID 
                   
               
               
                 NO: 17) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 83) 
                   
               
               
                   
                   
                   
                   
                 NO: 82) 
                   
                   
                   
               
               
                   
               
               
                 10-1121 
                 QVQLQESGPGLVKPPETLSLTCS 
                 DAYWS (SEQ 
                 WIRQSPGKRPE 
                 YVHHSGDTNY 
                 RVSFSLDTAKNEVSLKLVDLTAADSA 
                 ALHGKRIYGIVALGELF 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGASIN 
                 ID NO: 75) 
                 WVG 
                 NPSLKR 
                 IYFCAR 
                 TYFYMDV (SEQ ID 
                   
               
               
                 NO: 15) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 77) 
                   
               
               
                   
                   
                   
                   
                 NO: 76) 
                   
                   
                   
               
               
                   
               
               
                 10-1146 
                 QVQLVESGPGLVTPSETLSLTCT 
                 GRFWS (SEQ 
                 WIRQSPGRGLE 
                 YFSDTDRSEY 
                 RLTLSLDASRNQLSLKLKSVTAADSA 
                 AQQGKRIYGIVSFGEFF 
                 WGKGTAVTVSS 
               
               
                 (SEQ ID 
                 VSNGSVS 
                 ID NO: 87) 
                 WIG 
                 SPSLRS 
                 TYYCAR 
                 YYYYMDA (SEQ ID 
                   
               
               
                 NO: 19) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 89) 
                   
               
               
                   
                   
                   
                   
                 NO: 88) 
                   
                   
                   
               
               
                   
               
               
                 10-996 
                 QVQLQESGPGLVKPSETLSLTCS 
                 GRFWS (SEQ 
                 WIRQSPGRGLE 
                 YFSDTEKSNY 
                 RLTLSVDASKNQLSLKLNSVTAADSA 
                 TQQGKRIYGVVSFGEFF 
                 WGKGTAVTVSS 
               
               
                 (SEQ ID 
                 VSNGSVS 
                 ID NO: 63) 
                 WIG 
                 NPSLRS 
                 TYYCAR 
                 HYYYMDA (SEQ ID 
                   
               
               
                 NO: 11) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 65) 
                   
               
               
                   
                   
                   
                   
                 NO: 64) 
                   
                   
                   
               
               
                 GL (SEQ 
                 QVQLQESGPGLVKPSETLSLTCT 
                 SYYWS (SEQ 
                 WIRQPPGKGLE 
                 YIYYSGSTNY 
                 RVTISVDTSKNQFSLKLSSVTAADTA 
                 TQQGKRIYGVVSFGDYY 
                 WGKGTTVTVSS 
               
               
                 ID NO: 
                 VSGGSIS 
                 ID NO: 123) 
                 WIG 
                 NPSLKS 
                 VYYCAR 
                 YYYYMDV (SEQ ID 
                   
               
               
                 31) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 125) 
                   
               
               
                   
                   
                   
                   
                 NO: 124) 
                   
                   
                   
               
               
                 10-1341 
                 QVQLQESGPGLVKPSETLSVTCS 
                 NYYWT (SEQ 
                 WIRQSPGKGLE 
                 YISDRESATY 
                 RVVISRDTSTNQLSLKLNSVTPADTA 
                 ARRGQRIYGVVSFGEFF 
                 WGRGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGDSMN 
                 ID NO: 93) 
                 WIG 
                 NPSLNS 
                 VYYCAT 
                 YYYSMDV (SEQ ID 
                   
               
               
                 NO: 21) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 95) 
                   
               
               
                   
                   
                   
                   
                 NO: 94) 
                   
                   
                   
               
               
                 10-847 
                 QVQLQESGPGLVKPSETLSVTCS 
                 NYYWT (SEQ 
                 WIRQSPGKGLE 
                 YISDRASATY 
                 RVVISRDTSKNQLSLKLNSVTPADTA 
                 ARRGQRIYGVVSFGEFF 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGDSMN 
                 ID NO: 57) 
                 WIG 
                 NPSLNS 
                 VYYCAT 
                 YYYSMDV (SEQ ID 
                   
               
               
                 NO: 9) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 59) 
                   
               
               
                   
                   
                   
                   
                 NO: 58) 
                   
                   
                   
               
               
                 10-1074 
                 QVQLQESGPGLVKPSETLSVTCS 
                 NYYWT (SEQ 
                 WIRQSPGKGLE 
                 YISDRESATY 
                 RVVISRDTSKNQLSLKLNSVTPADTA 
                 ARRGQRIYGVVSFGEFF 
                 WGKGTTVTVSS 
               
               
                 (SEQ ID 
                 VSGDSMN 
                 ID NO: 69) 
                 WIG 
                 NPSLNS 
                 VYYCAT 
                 YYYSMDV (SEQ ID 
                   
               
               
                 NO: 13) 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 71) 
                   
               
               
                   
                   
                   
                   
                 NO: 70) 
                   
                   
                   
               
               
                 10- 
                 QVQLQESGPGLVKPSETLSVTCS 
                 NSYWT (SEQ 
                 WIRQSPGKGLE 
                 YISKSESANY 
                 RVVISRDTSKNQLSLKLNSVTPADTA 
                 ARHGQRIYGVVSFGEFF 
                 WGKGTTVTVSS 
               
               
                 1074GM 
                 VSGDSMN 
                 ID NO: 131) 
                 WIG 
                 NPSLNS 
                 VYYCAT 
                 TYYSMDV (SEQ ID 
                   
               
               
                 (SEQ ID 
                   
                   
                   
                 (SEQ ID 
                   
                 NO: 133) 
                   
               
               
                 NO: 129) 
                   
                   
                   
                 NO: 132) 
               
               
                   
               
            
           
           
               
            
               
                 IgL SEQUENCES 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 IMGT 
                 FWR1 
                 CDR1 
                 FWR2 
                 CDR2 
                 FWR3 
                 CDR3 
                 FWR4 
               
               
                   
               
               
                 GL (SEQ 
                 SYVLTQPPSVSVAPGQTARITCG 
                 NIGSKS (SEQ 
                 VHWYQQKPGQA 
                 DDS (SEQ 
                 DRPSGIPERFSGSNSGNTATLTISRV 
                 QVWDSSSDHPWV (SEQ 
                 FGGGTKLTVL 
               
               
                 ID NO: 
                 GN 
                 ID NO: 178) 
                 PVLVVY 
                 ID NO: 
                 EAGDEADYYC 
                 ID NO: 180) 
                   
               
               
                 32) 
                   
                   
                   
                 179) 
                   
                   
                   
               
               
                   
               
               
                 10-1369 
                 SSMSVSPGETAKITCGEK 
                 SIGSRA (SEQ 
                 VQWYQKKPGQP 
                 NNQ (SEQ 
                 DRPSGVPERFSASPDIEFGTTATLTI 
                 HIYDARRPTNWV (SEQ 
                 FDRGTTLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 181) 
                 PSLIIY 
                 ID NO: 
                 TNVEAGDEADYYC 
                 ID NO: 183) 
                   
               
               
                 NO: 24) 
                   
                   
                   
                 182) 
                   
                   
                   
               
               
                   
               
               
                 10-259 
                 SSMSVSPGETAKISCGKE 
                 SIGSRA (SEQ 
                 VQWYQQKSGQP 
                 NNQ (SEQ 
                 DRPSGVPERFSATPDFGAGTTATLTI 
                 HIYDARGGTNWV (SEQ 
                 FDRGATLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 184) 
                 PSLIIY 
                 ID NO: 
                 TNVEADDEADYYC 
                 ID NO: 186) 
                   
               
               
                 NO: 4) 
                   
                   
                   
                 185) 
                   
                   
                   
               
               
                   
               
               
                 10-303 
                 SDISVAPGETARISCGEK 
                 SLGSRA (SEQ 
                 VQWYQHRAGQA 
                 NNQ (SEQ 
                 DRPSGIPERFSGSPDSPFGTTATLTI 
                 HIWDSRVPTKWV (SEQ 
                 FGGGTTLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 187) 
                 PSLIIY 
                 ID NO: 
                 TSVEAGDEADYYC 
                 ID NO: 189) 
                   
               
               
                 NO: 6) 
                   
                   
                   
                 188) 
                   
                   
                   
               
               
                   
               
               
                 10-1121 
                 SFVSVAPGQTARITCGEE 
                 SLGSRS (SEQ 
                 VIWYQQRPGQA 
                 NNH (SEQ 
                 DRPSGIPERFSGSPGSTFGTTATLTI 
                 HIWDSRRPTNWV (SEQ 
                 FGEGTTLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 190) 
                 PSLIMY 
                 ID NO: 
                 TSVEAGDEADYYC 
                 ID NO: 192) 
                   
               
               
                 NO: 16) 
                   
                   
                   
                 191) 
                   
                   
                   
               
               
                   
               
               
                 10-410 
                 SFVSVAPGQTARITCGEE 
                 SLGSRS (SEQ 
                 VIWYQQRPGQA 
                 NNN (SEQ 
                 DRPSGIPERFSGSPGSTFGTTATLTI 
                 HIWDSRRPTNWV (SEQ 
                 FGEGTTLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 193) 
                 PSLIIY 
                 ID NO: 
                 TSVEAGDEADYYC 
                 ID NO: 195) 
                   
               
               
                 NO: 8) 
                   
                   
                   
                 194) 
                   
                   
                   
               
               
                   
               
               
                 10-1130 
                 SFVSVAPGQTARITCGEE 
                 SLGSRS (SEQ 
                 VIWYQQRPGQA 
                 NNN (SEQ 
                 DRPSGIPERFSGSPGSTFGTTATLTI 
                 HIWDSRRPTNWV (SEQ 
                 FGEGTTLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 196) 
                 PSLIIY 
                 ID NO: 
                 TSVEAGDEADYYC 
                 ID NO: 198) 
                   
               
               
                 NO: 18) 
                   
                   
                   
                 197) 
                   
                   
                   
               
               
                   
               
               
                 10-847 
                 SYVRPLSVALGETASISCGRQ 
                 ALGSRA (SEQ 
                 VQWYQHRPGQA 
                 NNQ (SEQ 
                 DRPSGIPERFSGTPDINFGTRATLTI 
                 HMWDSRSGFSWS (SEQ 
                 FGGATRLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 199) 
                 PILLIY 
                 ID NO: 
                 SGVEAGDEADYYC 
                 ID NO: 201) 
                   
               
               
                 NO: 10) 
                   
                   
                   
                   
                 200) 
                   
                   
               
               
                   
               
               
                 10-1074 
                 SYVRPLSVALGETARISCGRQ 
                 ALGSRA (SEQ 
                 VQWYQHRPGQA 
                 NNQ (SEQ 
                 DRPSGIPERFSGTPDINFGTRATLTI 
                 HMWDSRSGFSWS (SEQ 
                 FGGATRLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 202) 
                 PILLIY 
                 ID NO: 
                 SGVEAGDEADYYC 
                 ID NO: 204) 
                   
               
               
                 NO: 14) 
                   
                   
                   
                 203) 
                   
                   
                   
               
               
                   
               
               
                 10-1341 
                 SYVRPLSVALGETARISCGRQ 
                 ALGSRA (SEQ 
                 VQWYQHRPGQA 
                 NNQ (SEQ 
                 DRPSGIPERFSGTPDINFGTRATLTI 
                 HMWDSRSGFSWS (SEQ 
                 FGGATRLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 205) 
                 PILLIY 
                 ID NO: 
                 SGVEAGDEADYYC 
                 ID NO: 207) 
                   
               
               
                 NO: 22) 
                   
                   
                   
                 206) 
                   
                   
                   
               
               
                   
               
               
                 10-996 
                 SSLPLSVAPGATAKIACGEK 
                 SFASRA (SEQ 
                 VQWYQQKPGQA 
                 NNQ (SEQ 
                 DRPAGVSERFSGTPDVGFGSTATLTI 
                 HKWDSRSPLSWV (SEQ 
                 FGGGTQLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 208) 
                 PVLIIY 
                 ID NO: 
                 SRVEAGDEADYYC 
                 ID NO: 210) 
                   
               
               
                 NO: 12) 
                   
                   
                   
                 209) 
                   
                   
                   
               
               
                   
               
               
                 10-1146 
                 SSLPLSLAPGATAKIPCGEK 
                 SRGSRA (SEQ 
                 VQWYQQKPGQA 
                 NNQ (SEQ 
                 DRPAGVSERYSGNPDVAIGVTATLTI 
                 HYWDSRSPISWV (SEQ 
                 FGGWTQLTVL 
               
               
                 (SEQ ID 
                   
                 ID NO: 211) 
                 PTLIIY 
                 ID NO: 
                 SRVEAGDEAEYYC 
                 ID NO: 213) 
                   
               
               
                 NO: 20) 
                   
                   
                   
                 212) 
               
               
                   
               
               
                 KABAT 
                 FWR1 
                 CDR1 
                 FWR2 
                 CDR2 
                 FWR3 
                 CDR3 
                 FWR4 
               
               
                   
               
               
                 GL (SEQ 
                 SYVLTQPPSVSVAPGQTARITC 
                 GGNNIGSKSVH 
                 WYQQKPGQAPV 
                 DDSDRPS 
                 GIPERFSGSNSGNTATLTISRVEAGD 
                 QVWDSSSDHPWV (SEQ 
                 FGGGTKLTVL 
               
               
                 ID NO: 
                   
                 (SEQ ID NO: 
                 LVVY 
                 (SEQ ID 
                 EADYYC 
                 ID NO: 128) 
                   
               
               
                 32) 
                   
                 126) 
                   
                 NO: 127) 
                   
                   
                   
               
               
                   
               
               
                 10-1369 
                 SSMSVSPGETAKITC 
                 GEKSIGSRAVQ 
                 WYQKKPGQPPS 
                 NNQDRPS 
                 GVPERFSASPDIEFGTTATLTITNVE 
                 HIYDARRPTNWV (SEQ 
                 FDRGTTLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 104) 
                   
               
               
                 NO: 24) 
                   
                 102) 
                   
                 NO: 103) 
                   
                   
                   
               
               
                   
               
               
                 10-259 
                 SSMSVSPGETAKISC 
                 GKESIGSRAVQ 
                 WYQQKSGQPPS 
                 NNQDRPS 
                 GVPERFSATPDFGAGTTATLTITNVE 
                 HIYDARGGTNWV (SEQ 
                 FDRGATLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIIY 
                 (SEQ ID 
                 ADDEADYYC 
                 ID NO: 44) 
                   
               
               
                 NO: 4) 
                   
                 42) 
                   
                 NO: 43) 
                   
                   
                   
               
               
                   
               
               
                 10-303 
                 SDISVAPGETARISC 
                 GEKSLGSRAVQ 
                 WYQHRAGQAPS 
                 NNQDRPS 
                 GIPERFSGSPDSPFGTTATLTITSVE 
                 HIWDSRVPTKWV (SEQ 
                 FGGGTTLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 50) 
                   
               
               
                 NO: 6) 
                   
                 48) 
                   
                 NO: 49) 
                   
                   
                   
               
               
                   
               
               
                 10-1121 
                 SFVSVAPGQTARITC 
                 GEESLGSRSVI 
                 WYQQRPGQAPS 
                 NNHDRPS 
                 GIPERFSGSPGSTFGTTATLTITSVE 
                 HIWDSRRPTNWV (SEQ 
                 FGEGTTLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIMY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 80) 
                   
               
               
                 NO: 16) 
                   
                 78) 
                   
                 NO: 79) 
                   
                   
                   
               
               
                   
               
               
                 10-410 
                 SFVSVAPGQTARITC 
                 GEESLGSRSVI 
                 WYQQRPGQAPS 
                 NNNDRPS 
                 GIPERFSGSPGSTFGTTATLTITSVE 
                 HIWDSRRPTNWV (SEQ 
                 FGEGTTLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 56) 
                   
               
               
                 NO: 8) 
                   
                 54) 
                   
                 NO: 55) 
                   
                   
                   
               
               
                   
               
               
                 10-1130 
                 SFVSVAPGQTARITC 
                 GEESLGSRSVI 
                 WYQQRPGQAPS 
                 NNNDRPS 
                 GIPERFSGSPGSTFGTTATLTITSVE 
                 HIWDSRRPTNWV (SEQ 
                 FGEGTTLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 86) 
                   
               
               
                 NO: 18) 
                   
                 84) 
                   
                 NO: 85) 
                   
                   
                   
               
               
                   
               
               
                 10-847 
                 SYVRPLSVALGETASISC 
                 GRQALGSRAVQ 
                 WYQHRPGQAPI 
                 NNQDRPS 
                 GIPERFSGTPDINFGTRATLTISGVE 
                 HMWDSRSGFSWS (SEQ 
                 FGGATRLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LLIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 62) 
                   
               
               
                 NO: 10) 
                   
                 60) 
                   
                 NO: 61) 
                   
                   
                   
               
               
                   
               
               
                 10-1074 
                 SYVRPLSVALGETARISC 
                 GRQALGSRAVQ 
                 WYQHRPGQAPI 
                 NNQDRPS 
                 GIPERFSGTPDINFGTRATLTISGVE 
                 HMWDSRSGFSWS (SEQ 
                 FGGATRLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LLIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 74) 
                   
               
               
                 NO: 14) 
                   
                 72) 
                   
                 NO: 73) 
                   
                   
                   
               
               
                   
               
               
                 10-1341 
                 SYVRPLSVALGETARISC 
                 GRQALGSRAVQ 
                 WYQHRPGQAPI 
                 NNQDRPS 
                 GIPERFSGTPDINFGTRATLTISGVE 
                 HMWDSRSGFSWS (SEQ 
                 FGGATRLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LLIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 98) 
                   
               
               
                 NO: 22) 
                   
                 96) 
                   
                 NO: 97) 
                   
                   
                   
               
               
                   
               
               
                 10-996 
                 SSLPLSVAPGATAKIAC 
                 GEKSFASRAVQ 
                 WYQQKPGQAPV 
                 NNQDRPA 
                 GVSERFSGTPDVGFGSTATLTISRVE 
                 HKWDSRSPLSWV (SEQ 
                 FGGGTQLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIIY 
                 (SEQ ID 
                 AGDEADYYC 
                 ID NO: 68) 
                   
               
               
                 NO: 12) 
                   
                 66) 
                   
                 NO: 67) 
                   
                   
                   
               
               
                   
               
               
                 10-1146 
                 SSLPLSLAPGATAKIPC 
                 GEKSRGSRAVQ 
                 WYQQKPGQAPT 
                 NNQDRPA 
                 GVSERYSGNPDVAIGVTATLTISRVE 
                 HYWDSRSPISWV (SEQ 
                 FGGWTQLTVL 
               
               
                 (SEQ ID 
                   
                 (SEQ ID NO: 
                 LIIY 
                 (SEQ ID 
                 AGDEAEYYC 
                 ID NO: 92) 
                   
               
               
                 NO: 20) 
                   
                 90) 
                   
                 NO: 91) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                   
                 SEQ ID NOs 
               
            
           
           
               
               
               
            
               
                   
                 Variable Region 
                 CDRs 1-3 
               
            
           
           
               
               
               
               
               
            
               
                 Name 
                 Heavy chain (H) 
                 Light chain (L) 
                 H 
                 L 
               
               
                   
               
               
                 consensus 
                 SEQ ID NO: 1  
                 SEQ ID NO: 2  
                 SEQ ID NOs: 33-35  
                 SEQ ID NOs: 36-38  
               
               
                 10-259  
                 SEQ ID NO: 3  
                 SEQ ID NO: 4  
                 SEQ ID NOs: 39-41  
                 SEQ ID NOs: 42-44  
               
               
                 10-303  
                 SEQ ID NO: 5  
                 SEQ ID NO: 6  
                 SEQ ID NOs: 45-47  
                 SEQ ID NOs: 48-50  
               
               
                 10-410  
                 SEQ ID NO: 7  
                 SEQ ID NO: 8  
                 SEQ ID NOs: 51-53  
                 SEQ ID NOs: 54-56  
               
               
                 10-847  
                 SEQ ID NO: 9  
                 SEQ ID NO: 10  
                 SEQ ID NOs: 57-59  
                 SEQ ID NOs: 60-62  
               
               
                 10-996  
                 SEQ ID NO: 11  
                 SEQ ID NO: 12  
                 SEQ ID NOs: 63-65  
                 SEQ ID NOs: 66-68  
               
               
                 10-1074 
                 SEQ ID NO: 13  
                 SEQ ID NO: 14  
                 SEQ ID NOs: 69-71  
                 SEQ ID NOs: 72-74  
               
               
                 10-1121 
                 SEQ ID NO: 15  
                 SEQ ID NO: 16  
                 SEQ ID NOs: 75-77  
                 SEQ ID NOs: 78-80  
               
               
                 10-1130 
                 SEQ ID NO: 17  
                 SEQ ID NO: 18  
                 SEQ ID NOs: 81-83  
                 SEQ ID NOs: 84-86  
               
               
                 10-1146 
                 SEQ ID NO: 19  
                 SEQ ID NO: 20  
                 SEQ ID NOs: 87-89  
                 SEQ ID NOs: 90-92  
               
               
                 10-1341 
                 SEQ ID NO: 21  
                 SEQ ID NO: 22  
                 SEQ ID NOs: 93-95  
                 SEQ ID NOs: 96-98  
               
               
                 10-1369 
                 SEQ ID NO: 23  
                 SEQ ID NO: 24  
                 SEQ ID NOs: 99-101  
                 SEQ ID NOs: 102-104 
               
               
                 PGT-121 
                 SEQ ID NO: 25  
                 SEQ ID NO: 26  
                 SEQ ID NOs: 105-107 
                 SEQ ID NOs: 108-110 
               
               
                 PGT-122 
                 SEQ ID NO: 27  
                 SEQ ID NO: 28  
                 SEQ ID NOs: 111-113 
                 SEQ ID NOs: 114-116 
               
               
                 PGT-123 
                 SEQ ID NO: 29  
                 SEQ ID NO: 30  
                 SEQ ID NOs: 117-119 
                 SEQ ID NOs: 120-122 
               
               
                 GL 
                 SEQ ID NO: 31  
                 SEQ ID NO: 32  
                 SEQ ID NOs: 123-125 
                 SEQ ID NOs: 126-128 
               
               
                 10-1074GM 
                 SEQ ID NO: 129 
                 SEQ ID NO: 130 
                 SEQ ID NOs: 131-133 
                 SEQ ID NOs: 134-136 
               
               
                   
               
            
           
         
       
     
     Eleven new unique variants were expressed (Table 3) and demonstrated binding to YU-2 gp120 and gp140 by ELISA and surface plasmon resonance (SPR). Unless otherwise noted, the gp120 and gp140 proteins for these and other experiments were expressed in mammalian cells that can attach either a complex-type or a high-mannose N-glycan to a PNGS. The level of reactivity with gp120 differed between antibodies belonging to the PGT121 and 10-1074 groups, the latter exhibiting higher apparent affinities ( FIG.  3 A ) mainly due to slower dissociation from gp120/gp140 for the 10-1074-related antibodies ( FIG.  4 B ). 
     Example 3 PGT121 and 10-1074 Epitopes 
     Asn332 gp120  in the vicinity of the V3 loop stem was reported as critical for binding and viral neutralization by PGT121 (Nature 477(7365):466-470), thus we examined the role of V3 in antigen recognition by PGT121-like and 10-1074-like antibodies. ELISAs were performed using HXB2 gp120 “core” proteins that lack V1-V3 loops (gp120 core ) or retain a portion of V3 (2CC-core), and using a YU-2 gp120 mutant protein carrying a double alanine substitution in the V3 stem (gp120 GD324-5AA ). The tested antibodies showed decreased reactivity against variants lacking the V3 loop and gp120 GD324-5AA  when compared to intact YU-2 gp120, with the binding of 10-1074-group antibodies being the most affected ( FIGS.  5 A  and B). These results suggest that recognition by both antibody groups involves protein determinants in the vicinity of the V3 loop. None of the antibodies bound to overlapping peptides spanning V3, suggesting the targeted epitopes are discontinuous and/or require a particular conformation not achieved by isolated peptides ( FIG.  5 C ). 
     Asn332 gp120  (Asn337 gp120  in earlier numbering (J Proteome Res 7(4):1660-1674)) is the N-terminal residue of a potential N-glycosylation site (PNGS) defined as the sequence Asn-X-Ser/Thr. To determine whether Asn332 gp120  and/or its N-linked glycan are required for gp120 reactivity of the new PGT121- and 10-1074-group antibodies, we tested their binding to YU-2 gp120 N332A  by ELISA. The N332A substitution diminished the binding of PGT121 and all the new antibody variants, whereas their reactivity against a mutant gp120 lacking a nearby glycosylation site (gp120 NNT301-3AAA  mutant) was unchanged. To determine if a PNGS in addition to the Asn332 gp120  PNGS affects recognition by the new antibodies, we constructed a series of 11 double glycan mutants in which the N332A mutation in YU-2 gp120 was combined with mutation of PNGSs located between Asn262 gp120  and Asn406 gp120 . All of the PGT121-like and 10-1074-like antibodies bound to each of the double glycan mutants with comparable affinity as to that for gp120 N332A . 
     To compare overall glycan recognition by the PGT121- and 10-1074-like antibodies, we examined their binding to YU-2 gp120 treated with PNGase F, which cleaves both complex-type and high-mannose N-glycans. Because gp120 cannot be fully deglycosylated enzymatically unless it is denaturated, PNGase F treatment resulted in partial deglycosylation of natively-folded gp120 ( FIG.  6   ). Nevertheless, the reactivities of the two groups of antibodies differed in that partial deglycosylation of gp120 by PNGase F decreased the binding activity of all PGT121-like antibodies but none of the 10-1074-like antibodies ( FIG.  6 C ). Similar experiments conducted with YU-2 gp120 treated with Endo H, which cleaves high-mannose, but not complex-type, N-glycans, affected binding of 10-1074-like antibodies more than PGT121-like antibodies ( FIG.  6 D ). 
     An N-glycan microarray revealed that six of seven tested PGT121-like antibodies showed detectable binding to complex-type mono- or bi-antennary N-glycans terminating with galactose or α2-6-linked sialic acid but no detectable binding to high-mannose type glycans, corroborating and extending previous reports of no binding of PGT121-123 to high-mannose N-glycans and no competition by Man 4  and Man 9  dendrons for gp120 binding ( FIG.  7   ). In contrast, there was no detectable binding to protein-free glycans by 10-1074-like antibodies ( FIG.  7   ). Although PGT121-like antibodies bound to protein-free complex-type, but not high-mannose, N-glycans, PGT121-like antibodies retained binding to YU-2 gp120 produced in cells treated with kifunensine (gp120 kif ), a mannosidase inhibitor that results in exclusive attachment of high-mannose glycans to PNGSs ( FIG.  8 B ). Most of the PGT121-like antibodies exhibited a small, but reproducible, decrease in binding to gp120 kif . By contrast, 10-1074-like antibodies retained full binding to gp120 kif ( FIG.  8 B ). These results are consistent with the hypothesis that high-mannose, as well as complex-type, N-glycans can be involved in the epitope of PGT121-like antibodies. 
     Epitope mapping experiments were performed with two representative members of each group (PGT121 and 10-1369 for the PGT121-like group; 10-1074 and 10-996 for the 10-1074-like group) by competition ELISA. All four antibodies showed cross-competition, but PGT121 more modestly inhibited the binding of 10-996 and 10-1074 to gp120 than vice-versa. To further map the targeted epitopes, we used anti-gp120 antibodies that recognize the crown of the V3 loop ( FIG.  5   ), the CD4bs, the co-receptor binding site (CD4-induced; CD4i), a constellation of high-mannose N-glycans (2G12) (Journal of virology 76(14):7293-7305; Proc Natl Acad Sci USA 102(38):13372-13377)), or the V3 loop and N-linked glycans at positions 301 and 332 (PGT128). Anti-V3 crown antibodies inhibited the binding of PGT121 and 10-1369 but did not interfere with the binding of 10-996 and 10-1074. PGT128, and to a lesser extent 2G12, but not the CD4bs and CD4i antibodies, diminished the binding of all four antibodies to gp120. 
     Taken together, these data suggest that PGT121 clonal members recognize a site involving a protein determinant in the vicinity of the V3 loop and the Asn332 gp120 -associated glycan. However, the clone segregates into two families, the PGT121-like and 10-1074-like groups, which differ in their affinities for gp120 and in the role of glycans in epitope formation. 
     Example 4 Broad and Potent HIV Neutralization 
     To evaluate the neutralizing activity of the new PGT121 variants, we measured their ability to inhibit HIV infection of TZM-bl cells using 10 viral strains including R1166.c1, which lacks the PNGS at gp120 position 332. All PGT121 variants, including the 10-1074-like antibodies, neutralized 9 pseudoviruses and none neutralized the R1166.c1 control ( FIG.  1 A  and Table 4). Neutralizing activity correlated with affinity for the HIV spike, with the 10-1074 group showing slightly greater potencies than the PGT121 group ( FIG.  1 B  and  FIG.  4 C ). A representative germline version (GL) of the PGT121/10-1074 antibody clonotype failed to bind gp120/gp140 or neutralize any viruses in the panel, implying that somatic mutation is required for binding and neutralization. Pairing GL light chains with mutated 10-1074- or 10-996-group heavy chains failed to rescue binding or neutralization, suggesting that both mutated chains contribute to proper assembly of the antibody paratope. 
     Next assays were carried out to compared the neutralization activities of PGT121 and two 10-1074-like antibodies (10-996 and 10-1074) against an extended panel of 119 difficult-to-neutralize pseudoviruses (classified as tier-2 and tier-3) (Tables 4 and 5). 10-996 and 10-1074 showed neutralization potencies and breadth similar to PGT121 ( FIG.  1 C ,  FIG.  9   , and Tables 5 and 6). As anticipated, most viruses bearing amino acid changes at gp120 positions 332 and/or 334 (spanning the Asn332-X-Ser334/Thr334 PNGS) were resistant to neutralization (83.8% were resistant to PGT121, 100% were resistant to 10-1074 and 10-996). Mutation at this PNGS accounted for the majority of viruses resistant to neutralization (68.5% for 10-996, 72.5% for 10-1074 and 60.8% for PGT121) (Table 7). Comparable neutralization activities were observed for the IgG and Fab forms of PGT121 and 10-1074, suggesting that bivalency is not critical for their activity ( FIG.  1 D ). 
     To evaluate the potential role of complex-type N-glycans on the HIV envelope in neutralization by PGT121 and 10-1074, we produced high-mannose-only virions in two different ways: by assembling pseudoviruses in cells treated with kifunensine, which results in Man 9 GlcNAc 2  N-linked glycans, or by assembly in HEK 293S GnTI −/−  cells, which results in Man 5 GlcNAc 2  N-linked glycans. We found that PGT121 neutralized 2 of 3 kifunensine-derived PGT121-sensitive/10-1074-resistant strains equivalently to their counterparts produced in wildtype cells ( FIG.  8 C ). Two PGT121-sensitive/10-1074-sensitive viral strains produced in GnTI −/−  cells were equally as sensitive to PGT121 and 10-1074 as their counterparts produced in wildtype cells. Consistent with previous reports that complex-type N-glycans partially protect the CD4 binding site from antibody binding, the viruses produced in GnTI −/−  cells were more sensitive to CD4-binding site antibodies (NIH45-46 G54W  and 3BNC60) ( FIG.  8 D ). 
     Example 5 Newly-Transmitted HIV-1 
     We next examined the activity of PGT121 and 10-1074 against transmitted founder viruses by evaluating neutralization in a peripheral blood mononuclear cell (PBMC)-based assay using 95 clade B viruses isolated from a cohort of individuals who seroconverted between 1985 and 1989 (historical seroconverters, n=14) or between 2003 and 2006 (contemporary seroconverters, n=25) (51, 52). We compared PGT121 and 10-1074 with anti-CD4bs bNAbs and other bNAbs including VRC01, PG9/PG16, b12, 2G12, 4E10 and 2F5. Clustering analyses of neutralization activity showed segregation into two groups; the PGT121/10-1074 group contained the most active HIV neutralizers including the anti-CD4bs and PG9 antibodies (Table 8). Remarkably, 10-1074 showed exceptional neutralization potency on this clade B virus panel, exhibiting the greatest breadth at 0.1 μg/ml (67% of the 95 clade B viruses) of all bNAbs tested (Table 8). Although 10-1074 showed higher potency on contemporary clade B viruses than PGT121 (˜20-fold difference), both antibodies were more effective against historical than contemporary viruses ( FIG.  1 E  and  FIG.  10   ). 
     Example 6 Crystal Structures of PGT121, 10-1074 and GL 
     To investigate the structural determinants of the differences between PGT121-like and 1074-like antibodies, we solved crystal structures of the Fab fragments of PGT121, 10-1074 and a representative germline precursor (GL) at 3.0 Å, 1.9 Å and 2.4 Å resolution, respectively (Table 9). Superimposition of the heavy and light chain variable domains (V H  and V L ) among the three Fabs showed conservation of the backbone structure, with differences limited to small displacements of the CDRH3 and CDRL3 loops of the affinity-matured Fabs relative to GL (Table 10). 
     An unusual feature shared by the antibodies is their long (25 residues) CDRH3 loop, which forms a two-stranded anti-parallel β-sheet extending the V H  domain F and G strands. In each Fab, the tip of the extended CDRH3 loop primarily contains non-polar residues. A similar structural feature was observed for the CDRH3 of PGT145, a carbohydrate-sensitive antibody whose epitope involves the gp120 V1V2 loop. However, the extended two-stranded β-sheet of PGT145&#39;s CDRH3 contains mostly negatively-charged residues, including two sulfated tyrosines at the tip. Aligning V H -V L  of PGT121 and PGT145 (Table 10) shows that CDRH3 PGT145  extends past CDRH3 PGT121  and that its tip and V H  domain are aligned, whereas the CDRH3s of PGT121, 10-1074 and GL tilt towards V L . The tilting of CDRH3PGT121/CDRH3 10-1074 /CDRH3 GL  towards V L  opens a cleft between CDRH2 and CDRH3, a feature not shared by related antibodies. 
     PGT121 and 10-1074 are highly divergent with respect to GL and each other (of 132 residues, PGT121 VH  differs from 10-1074 VH  and GL VH  by 36 and 45 residues, respectively, and 10-1074 VH  and GL VH  differ by 29). The majority of the PGT121/10-1074 differences are located in the CDR VH  loops and CDRL3. Interestingly, six substitutions in CDRH3 (residues 100d, 100f, 100h, 100j, 1001, 100n) alternate such that every second residue is substituted, causing resurfacing of the cleft between CDRH2 and CDRH3 that results from CDRH3 tilting towards V L . This region likely contributes to the different fine specificities of PGT121 and 10-1074. Five other solvent-exposed substitutions in heavy chain framework region 3 (FWR3 HC ) (residues 64, 78, 80-82; strands D and E) are potential antigen contact sites given that framework regions in HIV antibodies can contact gp120. Other differences that may contribute to fine specificity differences include a negative patch on PGT121 in the vicinity of Asp56 HC  not present in 10-1074 or GL (Ser56 HC  in 10-1074 and GL) and positive patches on the CDRL1 and CDRL3 surface not found on the analogous surface of GL. 
     Somatic mutations common to PGT121 and 10-1074 may be involved in shared features of their epitopes. The heavy chains of PGT121 and 10-1074 share only three common mutations (of 36 PGT121-GL and 29 10-1074-GL differences). In contrast, PGT121 and 10-1074 share 18 common light chain mutations (of 37 PGT121-GL and 36 10-1074-GL differences), including an insertion in light chain FWR3 that causes bulging of the loop connecting strands D and E, and the substitution of Asp50 LC -Asp51 LC  in CDRL2 GL  to Asn50 LC -Asn51 LC  in both PGT121 and 10-1074, resulting in a less negatively-charged surface. The large number of common substitutions introduced into LC PGT121  and LC 10-1074  (approximately 50% of LC substitutions) point to CDRL1, CDRL2 and FWR2 LC  as potential contact regions for epitopes shared by PGT121 and 10-1074. 
     Next, comparisons were made with the structure of PGT128, which recognizes Asn332 gp120 - and Asn301 gp120 -linked glycans and V3 and was solved as a complex with an outer domain/mini-V3 loop gp120 expressed in cells that cannot produce complex-type N-glycan-modified proteins. Unlike the CDRH3 loops of PGT121 and 10-1074, PGT128 CDRH3  is not tilted towards PGT128 VL , and CDRH3 PGT128  does not include a two-stranded β-sheet. In addition, CDRH3 PGT128  (18 residues) is shorter than the CDRH3s of PGT121 and 10-1074 (24 residues), whereas CDRH2 PGT128  contains a six-residue insertion not found in PGT121 or 10-1074. Due to these differences, CDRH2 is the most prominent feature in PGT128, whereas CDRH3 is most prominent in PGT121 and 10-1074. CDRH2 PGT128  and CDRL3 PGT128  together recognize Man 8/9  attached to Asn332 gp120 , and CDRH3 PGT128  contacts the V3 loop base. This mode of gp120 recognition is not possible for PGT121 and 10-1074 because the structural characteristics of their CDRH2 and CDRH3 loops differ significantly from those of PGT128, consistent with the ability of PGT128, but not PGT121 and 10-1074 ( FIG.  7   ), to recognize protein-free high-mannose glycans. 
     Example 7 Crystal Structure of PGT121-Glycan Complex 
     A 2.4 Å resolution structure of PGT121 associated with a complex-type sialylated bi-antennary glycan was solved (Table 9) using crystals obtained under conditions including NA2, a complex-type asialyl bi-antennary glycan ( FIG.  7   ). Surprisingly, the glycan bound to PGT121 in our crystal structure was not NA2, but rather a complex-type N-glycan from a neighboring PGT121 Fab in the crystal lattice; specifically the N-glycan attached to Asn105 HC . The glycan identity is evident because there was electron density for the glycosidic linkage to Asn105 HC  and for a terminal sialic acid on the Manα1-3Man antenna (the galactose and sialic acid moieties of Manα1-6Man antenna were unresolved). The composition of the bound glycan corresponds to a portion of the α2-6-sialylated A2(2-6) glycan that was bound by PGT121 in microarray experiments ( FIG.  7   ) and to the expected sialyl linkage on complex-type N-glycans attached to PNGS on proteins expressed in HEK293T cells. Although the V H -V L  domains of this structure (“liganded” PGT121) superimpose with no significant differences onto the V H -V L  domains of the PGT121 structure with no bound N-glycan (“unliganded” PGT121) (Table 10), the elbow bend angle (angle between the V H -V L  and C H 1-C L  pseudo-dyads) differs between the structures. This difference likely reflects flexibility that allows the Fab to adopt variable elbow bend angles depending upon crystal lattice forces. 
     Given that we observed binding of complex-type N-glycan in one crystal structure (the “liganded” PGT121 structure) but not in another structure (the “unliganded” PGT121 structure), we estimate that the affinity of PGT121 for complex-type N-glycan not attached to gp120 is in the range of the concentration of PGT121 in crystals (˜10 mM). If we assume that the K D  for binding isolated glycan is in the range of 1-10 mM, comparable to the 1.6 mM K D  derived for PG9 binding to Man 5 GlcNAc 2 -Asn, then the K D  for PGT121 binding of isolated glycan represents only a minor contribution to the affinity of PGT121 for gp120, which is in the nM range ( FIG.  4 A ). 
     The glycan in the “liganded” PGT121 structure interacts exclusively with the V H  domain and makes extensive contacts with residues in all three CDRs (buried surface area on PGT121 HC =600 Å 2 ). Contacts include 10 direct and 18 water-mediated hydrogen bonds (Table 11) with 9 amino acids anchoring the glycan between the N-acetylglucosamine moiety linked to the branch-point mannose and the terminal sialic acid on the 1-3-antenna. Several contacts with PGT121 are made by this sialic acid, including three direct hydrogen bonds with PGT121 residues Asp31 HC  and His97 HC  in addition to water-mediated hydrogen bonds with Asp31 HC . The sialic acid also contributes to a water-mediated intra-glycan hydrogen bond network. The direct contacts with sialic acid may explain the stronger binding of PGT121 to the sialylated A2(2-6) glycan than to the asialylated NA2 glycan in our glycan microarray analysis ( FIG.  7   ). Extensive water-mediated protein contacts established by the N-acetylglucosamine and galactose moieties of the 1-3-antenna could explain the binding observed for asialylated mono- and bi-antennary glycans to PGT121 ( FIG.  7   ). 
     Six of the residues contributing direct or likely amino acid side chain contacts to the glycan (Ser32 HC-CDRH1 , Lys53 HC-CDRH2 , Ser54 HC-CDRH2 , Asn58 HC-CDRH2 , His97 HC-CDRH3 , Thr1001 HC-CDRH3 ) differ from those on 10-1074. (Tyr32 HC-CDRH1 , Asp53 HC-CDRH2 , Arg54 HC-CDRH2 , Thr58 HC-CDRH2 , Arg97 HC-CDRH3 , Tyr1001 HC-CDRH 3), and are highly conserved among PGT121-like, but not 10-1074-like, antibodies. The 10-1074 residues lack the corresponding functional groups to make the observed glycan contacts or have bulky side chains that would cause steric clashes. Four of these residues also differ from those on GL (Tyr32 HC-CDRH1 , Tyr53 HC-CDRH2 , Gln97 HC-CDRH3 , Tyr1001 HC-CDRH3 ), suggesting that the lack of binding of 10-1074-like antibodies and GL to protein-free complex-type glycans in our glycan microarrays results from missing hydrogen bonds and/or steric clashes (e.g., His97 PGT121  versus Arg97 10-1074 ; Thr1001 PGT-121  versus Tyr1001 10-1074 ). As the majority of sequence differences between PGT121 and 10-1074 cluster in the CDRH loops, specifically to the surface of the cleft between CDRH2 and CDRH3 where we observe the bound complex-type N-glycan, differential recognition of complex-type glycans on gp120 may account for some or all of the differences in their fine specificity observed. 
     Example 8 Substitution of Glycan-Contacting Antibody Residues Affects Neutralization 
     To evaluate the contributions of complex-type N-glycan contacting residues identified from the “liganded” PGT121 structure, we generated two mutant antibodies designed to exchange the complex-type glycan-contacting residues between PGT121 and 10-1074: a 10-1074 IgG with PGT121 residues (six substitutions in IgH Y32S, D53K, R54S, T58N, R97H, Y1001T) and a PGT121 IgG with reciprocal substitutions. The “glycomutant” antibodies (10-1074 GM  and PGT121 GM ) exhibited near-wildtype apparent affinity for YU-2 gp120/gp140 as measured by SPR ( FIG.  2 A ), demonstrating that the substitutions did not destroy binding to an envelope spike derived from a viral strain neutralized by both PGT121 and 10-1074 ( FIG.  1 A ). The fact that PGT121 complex-type N-glycan contacting residues can be accommodated within the 10-1074 background without destroying binding to a gp120/gp140 bound by both wildtype antibodies implies overall similarity in antigen binding despite fine specificity differences. 
     Unlike wildtype PGT121, PGT121 GM  showed no glycan binding in microarray experiments, confirming that 10-1074 residues at the substituted positions are not compatible with protein-free glycan binding ( FIG.  2 B ) and supporting the suggestion that residues contacting the glycan in the “liganded” PGT121 structure are involved in recognition of complex-type glycans in the microarrays. 10-1074 GM  also showed no binding to protein-free glycans ( FIG.  2 B ), indicating the involvement of residues in addition to those substituted in creating the binding site for a protein-free complex-type N-glycan. 
     Next, a TZM-bl-based assay was used to compare neutralization of the wildtype and “glycomutant” antibodies. We tested 40 viral strains including strains differentially resistant to PGT121 or 10-1074 and strains sensitive to both wildtype antibodies ( FIG.  2 C  and Table 12). Consistent with the binding of PGT121 GM  and 10-1074 GM  to purified YU-2 envelope proteins, both mutants neutralized the YU-2 virus; however, 64% of the PGT121-sensitive strains were resistant to PGT121 GM  ( FIG.  2 C , and Table 12) suggesting that the glycan-contacting residues identified in the “liganded” PGT121 structure are relevant to the neutralization activity of PGT121. Conversely, 10-1074 GM  exhibited a higher average potency than wildtype 10-1074 against the 10-1074-sensitive strains ( FIG.  2 C  and Table 12), including potency increases of &gt;3-fold against four 10-1074-sensitive strains (WITO4160.33, ZM214M.PL15, Ce1172_H1, and 3817.v2.c59). In general, the PGT121 substitutions into 10-1074 did not confer sensitivity to 10-1074 GM  upon PGT121-sensitive/10-1074-resistant strains, however two of these strains (CNE19 and 62357_14_D3_4589) became sensitive to 10-1074 GM  (IC 50 s=0.19 μg/ml and 40.8 μg/ml, respectively). Interestingly, these are the only PGT121-sensitive/10-1074-resistant strains that include an intact Asn332 gp120 -linked PNGS. The other PGT121-sensitive/10-1074-resistant strains lack the Asn332 gp120 -linked glycan and are resistant to PGT121 GM  and 10-1074 GM , implying that their sensitivity to wildtype PGT121 involve a nearby N-glycan and/or compensation by protein portions of the epitope. Although a dramatic gain of function was observed only for 10-1074 GM  against one strain (CNE19), this result, together with the general improvement observed for 10-1074 GM  against 10-1074-sensitive strains ( FIG.  2 C ), is consistent with the interpretation that the crystallographically-identified glycan-contacting residues can transfer PGT121-like recognition properties to 10-1074 in some contexts and/or affect its potency in others. In addition, the loss of neutralization activity for PGT121 GM  against PGT121-sensitive strains demonstrates that neutralization activity of PGT121 involves residues identified as contacting complex-type N-glycan in the “liganded” PGT121 structure. 
     Results 
     PGT121 is a glycan-dependent bNAb that was originally identified in the serum of a clade A-infected donor in a functional screen yielding only two clonally-related members. gp140 trimers were used as “bait” for single cell sorting to isolate 29 new clonal variants of. The PGT121 clonal family includes distinct groups of closely-related antibodies; the PGT121- and 10-1074-groups. The results suggest that the epitopes of both groups involve the PNGS at Asn332 gp120  and the base of the V3 loop. The PGT121-like and 10-1074-like antibody groups differ in amino acid sequences, gp120/gp140 binding affinities, and neutralizing activities, with the 10-1074-like antibodies being completely dependent for neutralization upon an intact PNGS at Asn332 gp120 , whereas PGT121-like antibodies were able to neutralize some viral strains lacking the Asn332 gp120  PNGS. 
     A notable difference between the two antibody groups is that the PGT121-like antibodies bound complex-type N-glycans in carbohydrate arrays, whereas the 10-1074-like antibodies showed no detectable binding to any of the protein-free N-glycans tested ( FIG.  7   ). Protein-free glycan binding by anti-HIV antibodies is not always detectable; e.g., although PG9 recognizes a gp120-associated high-mannose glycan, no binding to protein-free glycans was detected in microarrays. Thus although a positive result in a glycan microarray implies involvement of a particular glycan in an antibody epitope, a negative result does not rule out glycan recognition. For example, although not detectable in the glycan microarray experiments, high-mannose glycans may be involved in the PGT121 epitope, consistent with binding and neutralization of high-mannose-only forms of gp120 protein and virions ( FIG.  8   ). 
     The molecular basis for the differences between PGT121, 10-1074 and their GL progenitor was revealed in part by their crystal structures. The finding that the majority of light chain somatic mutations are shared between PGT121 and 10-1074, whereas mutations in the heavy chains differ, suggests that the light chain contacts shared portions of the gp120 epitope and the heavy chain recognizes distinct features. All three antibodies exhibit an extended CDRH3 with a non-polar tip that may allow accessing of cryptic epitopes. Differences in the antigen-binding site of the two mature Fabs were mainly localized to a cleft between CDRH2 and the extended CDRH3. Interestingly, the putative antigen-binding cleft between CDRH2 and CDRH3 was also found in a representative germline progenitor of PGT121 and 10-1074. 
     Structural information was obtained concerning glycan recognition by PGT121-like antibodies from a crystal structure in which a complex-type sialylated N-glycan attached to a V H  domain residue interacted with the combining site of a neighboring PGT121 Fab. Several features of the “liganded” PGT121 structure suggest it is relevant for understanding the recognition of complex-type N-glycans on gp120 by PGT121-like antibodies. First, the glycan in the structure corresponds to the α2-6 sialylated glycan A2(2-6) PGT121 binds in microarrays ( FIG.  7   ). Second, the glycan interacts with PGT121 using the cleft between CDRH3 and CDRH2 that was suggested by structural analyses to be involved in epitope recognition, potentially explaining the unusual tilting of CDRH3 towards V L  in the PGT121 and 10-1074 structures. Third, most of the V H  residues identified as interacting with the glycan differ between PGT121 and 10-1074, rationalizing different binding profiles in glycan microarrays and potentially explaining the different fine specificities revealed in protein binding experiments. Fourth, swapping crystallographically-identified glycan contact residues between PGT121 and 10-1074 in part transferred their properties: PGT121 GM , like 10-1074, did not bind to protein-free glycans, but both PGT121 GM  and 10-1074 GM  preserved near wildtype binding to purified YU-2 gp120/gp140. Although PGT121 GM  retained the ability to neutralize some viral strains that were neutralized by wildtype PGT121 and 10-1074, it failed to neutralize strains that are PGT121-sensitive/10-1074-resistant, demonstrating that the glycan-binding motif is essential for the neutralizing activity of PGT121 against 10-1074-resistant strains. For the reciprocal swap, the neutralization potency of 10-1074 GM  was increased or unaffected relative to 10-1074, and in one case, 10-1074 GM  potently neutralized a PGT121-sensitive/10-1074-resistant strain, consistent with transfer of the crystallographically-identified glycan motif and the hypothesis that the epitopes of PGT121- and 10-1074-like antibodies are related. In analyses of gp120 sequences from strains for which PGT121 neutralization data are available, other than a correlation with the PNGS at Asn332gp120 for viruses sensitive to PGT121-like and 10-1074-like antibodies, no clear pattern of PNGS usage emerges for the different categories of viral strains (PGT121-sensitive/10-1074-sensitive, PGT121-sensitive/10-1074-resistant, PGT121-resistant/10-1074-sensitive) except that the 10-1074-resistant strains generally lack the Asn332gp120-associated PNGS. 
     Example 9 Passive Transfer of Anti-HIV-1 Neutralizing mAbs In-Vivo 
     Five isolated potent and broadly acting anti-HIV neutralizing monoclonal antibodies were administered to rhesus macaques and challenged them intrarectally 24 h later with either of two different SHIVs. By combining the results obtained from 60 challenged animals, the protective neutralization titer in plasma preventing virus acquisition in 50% of the exposed monkeys was approximately 1:100. 
     Animal Experiments 
     The macaques used in this study were negative for the MHC class I Mamu-A*01 allele. 
     Construction of the R5-Tropic SHIVDH12-V3AD8 PCR mutagenesis, with primers corresponding to the 5′ and 3′ halves of the SHIVAD8EO (PNAS 109, 19769-19774 (2012)) gp120 V3 coding region (forward primer: AGAGCATTTTATACAACAGGAGACATAATAGGAGATATAAGACAAGCACATTGCAA CATTAGTAAAGTAAAATGGC (SEQ ID NO: 214) and reverse primer: TCCTGGTCCTATATGTATACTTTTCCTTGTATTGTTGTTGGGTCTTGTACAATTAATTT CTACAGTTTCATTC (SEQ ID NO: 215)), was employed to introduce these V3 sequences into the genetic background of the pSHIVDH12-CL7 molecular clone ( J. of Virology  78, 5513-5519 (2004)), using Platinum PFX DNA polymerase (Invitrogen). Following gel purification, the PCR product was treated with T4 polynucleotide kinase (GibcoBRL) and blunt-end ligated to create pSHIVDH12-V3AD8, which was used to transform competent cells. 
     Viruses 
     Virus stocks were prepared by first transfecting 293T cells with the SHIVAD8EO or SHIVDH12-V3AD8 molecular clones using Lipofectamine 2000 (Invitrogen, Carlsbad, Calif.). Culture supernatants were collected 48 h later and aliquots stored at −80° C. until use. Concanavalin A-stimulated rhesus PBMCs (2×10 6  cells in 500 μl) were infected with transfected cell supernatants by spinoculation (J. of Virology 74, 10074-10080 (2000)) for 1 h, mixed with the same number/volume of activated PBMC, and cultures were maintained for at least 12 days with daily replacement of culture medium. Samples of supernatant medium were pooled around the times of peak RT production to prepare individual virus stocks. 
     Antibodies 
     Eleven monoclonal antibodies (VRC01, NIH45-46, 45-46G54W, 45-46m2, 3BNC117, 12A12, 1NC9, and 8ANC195, 10-1074, PGT121, and PGT126) were isolated and produced. DEN3, a dengue virus NS1-specific human IgG1 monoclonal antibody (PNAS 109, 18921-18925 (2012)), or control human IgG (NIH Nonhuman Primate Reagent Resource) were used as the negative control antibodies in this study. The monoclonal antibodies selected for pre-exposure passive transfer were administered intravenously 24 h before virus challenge. 
     Quantitation of Plasma Viral RNA Levels. 
     Viral RNA levels in plasma were determined by real-time reverse transcription-PCR (ABI Prism 7900HT sequence detection system; Applied Biosystems). 
     Antibody Concentrations in Plasma. 
     The concentrations of administered monoclonal antibodies in monkey plasma were determined by enzyme-linked immunosorbent assay (ELISA) using recombinant HIV-1JRFL gp120 (Progenics Pharmaceuticals) or HIVIIIB (Advanced Biotechnology inc) (J. of Virology 75, 8340-8347 (2001)). Briefly, microtiter plates were coated with HIV-1 gp120 (2 μg/ml) and incubated overnight at 4° C. The plates were washed with PBS/0.05% Tween-20 and blocked with 1% (vol/vol) BSA. After blocking, serial dilution of antibodies or plasma samples were added to the plate and incubated for 1 h at room temperature. Binding was detected with a goat anti-human IgG F(ab)2 fragments coupled to alkaline phosphatase (Pierce) and visualized with SIGMAFAST OPD (Sigma-Aldrich). The decay half-lives of neutralizing monoclonal antibodies were calculated by a single-exponential decay formula based on the plasma concentrations beginning on day 5 or day 7 post antibody administration (J. of Virology 84, 1302-1313 (2010)). 
     Neutralization Assays. 
     The in vitro potency of each mAb and the neutralization activity present in plasma samples collected from rhesus macaques were assessed by two types of neutralization assays; 1) TZM-bl entry assay with pseudotyped challenge virus (AIDS Res Hum Retroviruses 26, 89-98 (2010)) or 2) a 14 day PBMC replication assay with replication competent virus (J. of virology 76, 2123-2130 (2002)). For the TZM-bl assay, serially diluted mAb or plasma samples were incubated with pseudotyped viruses, expressing env gene derived from SHIVAD8EO or SHIVDH12-V3AD8 and prepared by cotransfecting 293T cells with pNLenv1 and pCMV vectors expressing the respective envelope proteins (J. of Virology 84, 4769-4781 (2010)). The 50% neutralization inhibitory dose (IC50) titer was calculated as the dilution causing a 50% reduction in relative luminescence units (RLU) compared with levels in virus control wells after subtraction of cell control RLU (J. of Virology 84, 1439-1452 (2010)). The neutralization phenotype (tier levels) of the SHIVDH12-V3AD8 molecular clone was determined by TZM-bl cell assay using plasma samples from a cohort study, which exhibit a wide range of neutralizing activities against subtype B HIV-1 isolates (J. of General Virology 91, 2794-2803 (2010)). 
     Determinations of Animal Protective Titers and Statistical Analyses. 
     Calculation of the neutralizing titer in plasma against each R5 SHIV, resulting in the prevention of virus acquisition of 50 or 80% of the virus-challenged animals, was performed using the method of Reed and Muench (Am J Hyg 27, 493-497 (1938)). One significant outlier animal (DEW7) was omitted from the calculation. Probit regression was used to model the relationship between the titers in plasma required to confer sterilizing immunity in vivo using all 60 passively immunized monkeys (Cambridge University Press, Cambridge, England, ed. 3rd, 2007), with p-values from this model based on Likelihood ratio Tests. Plasma titers needed for different levels of in vivo protection (33%, 50%, 80%, 90%, and 95%) were determined from the probit model estimates and the method of bootstrapping was used to construct 90% confidence intervals. Results: 
     SHIVDH12-V3AD8, like SHIVAD8EO, possesses Tier 2 anti-HIV-1 neutralization sensitivity properties (Table 13). Rhesus macaques inoculated intravenously or intrarectally with SHIVDH12-V3AD8 exhibited peak viremia ranging from 105 to 107 viral RNA copes/ml of plasma at weeks 2 to 3 post infection (PI). In most SHIVDH12-V3AD8 infected animals, plasma viral loads decline to background levels between weeks 8 to 20 PI. 
     The neutralization sensitivity of SHIVAD8EO to 11 recently reported broadly reacting anti-HIV-1 mAbs was initially determined in the TZM-bl assay system ( FIGS.  11 A  and B). Eight of these antibodies, VRC01, NIH45-46 (23), 45-46G54W, 45-46m2, 3BNC117, 12A12, 1NC9, and 8ANC195 targeted the gp120 CD4 bs (Science 333, 1633-1637 (2011)) and three, 10-1074, PGT121, and PGT126 (Nature 477, 466-470 (2011)), were dependent on the presence of the HIV-1 gp120 N332 glycan. When tested against SHIVAD8EO, all three glycan-dependent mAbs exhibited greater potency than the CD4 bs mAbs ( FIG.  11    A). The IC50 values for the three mAbs targeting the gp120 N332 glycan ranged from 0.09 to 0.15 μg/ml. The CD4 bs mAbs exhibited a much broader range (0.14 to 6.34 μg/ml) of IC50 neutralizing activity with 3BNC117 being the most potent. A similar hierarchy (glycan-dependent &gt;CD4 bs dependent) of neutralizing mAb potency was also observed with SHIVDH12-V3AD8, but the neutralizing activity was distributed across a much wider (&gt;100 fold) range compared to the IC50 values observed for SHIVAD8EO ( FIG.  11 B ). SHIVDH12-V3AD8 was somewhat more sensitive to the glycan targeting mAbs and more resistant to the CD4 bs neutralizing mAbs than SHIVAD8EO. 
     Based on the results shown in  FIG.  11   , five neutralizing mAbs were selected for a pre-exposure passive transfer study: VRC01, because it was the first CD4bs NAb of the newly isolated broadly acting NAbs to be characterized; the CD4 bs mAbs 45-46m2 and 3BNC117, both of which exhibited strong neutralizing activity against SHIVAD8EO and SHIVDH12-V3AD8; and the gp120 N332 glycan-dependent mAbs, PGT121 and 10-1074. 
     The protocol for passive transfer experiments was to administer decreasing amounts of neutralizing mAbs intravenously and challenge animals intrarectally 24 h later. The goal was to block virus acquisition, coupled with the knowledge that repeated administrations of humanized anti-HIV mAbs to individual macaques could reduce their potency and/or possibly induce anaphylactic responses, a SHIV challenge dose of sufficient size to establish an in vivo infection following a single inoculation was chosen. In this regard, we had previously conducted intrarectal titrations of SHIVAD8 in rhesus monkeys and reported that the inoculation of 1×103 TCID50, determined by endpoint dilution in rhesus macaque PBMC, was equivalent to administering approximately 3 animal infectious doses50 (AID50) (J. of virology 86, 8516-8526 (2012)). In fact, single intrarectal inoculations of 3 AID50 have resulted in the successful establishment of infection in 10 of 10 rhesus macaques with SHIVAD8EO or SHIVDH12-V3AD8. 
     As a control for the first passive transfer experiment, an anti-dengue virus NS1 IgG1 mAb was administered intravenously to animals, which were challenged with SHIVAD8EO 24 h later. Both monkeys (ML1 and MAA) rapidly became infected, generating peak levels of plasma viremia at week 2 PI. VRC01 was the first anti-HIV-1 neutralizing mAb tested for protection against virus acquisition and was administered to two macaques at a dose of 50 mg/kg. One (DEGF) of the two inoculated macaques was completely protected from the SHIVAD8EO challenge, with no evidence of plasma viremia or cell-associated viral DNA over a 45 week observation period. The other recipient of 50 mg/kg VRC01 (DEH3) became infected, but peak plasma viremia was delayed until week 5 PI. Two additional macaques administered lower amounts (20 mg/kg) of VRC01 were not protected from the SHIVAD8EO challenge. These results are summarized in Table 13. 
     Examined next, the protective properties of PGT121 against a SHIVAD8EO challenge. PGT121 was one of the most potent glycan targeting neutralizing mAbs measured in the TZM-bl assay ( FIG.  11   ). Based on the results obtained with VRC01, in vivo PGT121 mAb titration at 20 mg/kg was chosen to begin with. The two challenged monkeys (KNX and MK4) resisted the SHIVAD8EO challenge. When lower amounts (viz. 5 mg/kg, 1 mg/kg, or 0.2 mg/kg) of PGT121 were administered, 1 of 2, 2 of 2, and 0 of 2 animals, respectively, were protected (Table 13). 
     The capacity of VRC01 and PGT121 mAbs to block SHIVDH12-V3AD8 acquisition was similarly evaluated (Table 13). The results obtained with VRC01 were comparable to those observed with the SHIVAD8EO challenge: 1 of 2 recipients of 30 mg/kg was protected from the establishment of a SHIVDH12-V3AD8 infection. The PGT121 mAb was considerably more potent than VRC01 in preventing SHIVDH12-V3AD8 acquisition: 2 of 2 recipients of 0.2 mg/kg PGT121 resisted infection. PGT121 also appeared to be somewhat more effective in preventing SHIVDH12-V3AD8 versus SHIVAD8EO in vivo infections (Table 13). This result is consistent with the 8-fold difference in IC50 values for PGT121 for neutralizing the two SHIVs in in vitro assays ( FIG.  11   ). 
     The results of passively transferring 10-1074, 3BNC117, or 45-46m2 neutralizing mAbs to rhesus monkeys, followed by a challenge with either SHIVAD8EO or SHIVDH12-V3AD8, are summarized in Table 13. The 10-1074 mAb potently blocked the in vivo acquisition of both SHIVs. The CD4bs 3BNC117 and 45-46m2 mAbs were selected for passive transfer to macaques based on their IC50 values against both SHIVs in the in vitro neutralization experiments shown in  FIG.  11   . 3BNC117 successfully blocked SHIVAD8EO infection in 2 of 2 monkeys at 5 mg/kg but not in 2 other animals given a dose of 1 mg/kg (Table 13). This was similar to the results observed when the same amounts of 3BNC117 were administered to macaques challenged with SHIVDH12-V3AD8: 1 of 2 became infected at 5 mg/kg; 1 of 2 became infected at 1 mg/kg. 
     Plasma samples collected at various times from passively transferred macaques were analyzed by HIV-1 gp120 ELISA to determine neutralizing mAb concentrations. In general, the plasma concentrations of each mAb at the time of challenge (24 h following antibody administration) correlated with the dose of antibody administered (Table 13). 
     The relationships of plasma mAb concentrations to in vivo protection are shown in  FIG.  12   . Of the 5 neutralizing mAbs evaluated, PGT121 was clearly the most effective against both viruses, with SHIVDH12-V3AD8 exhibiting somewhat greater sensitivity to this mAb (2 of 2 monkeys protected at a plasma concentration of 0.2 μg/ml). In contrast, a plasma concentration of nearly 400 μg/ml of VRC01 was required to protect 1 of 2 animals against the same SHIVDH12-V3AD8 challenge virus (Table 13). The most potent CD4 bs mAb administered to macaques in this study, 3BNC117, was approximately 6 to 10-fold more effective than VRC01 in preventing the acquisition of either SHIV ( FIG.  12   , Table 13). 
     The calculated half lives of PGT121, 10-1074, 3BNC117, and VRC01 mAbs were quite similar: 3.5 days, 3.5 days, 3.3 days, and 3.1 days, respectively. In contrast, the half-life of 45-46m2 was extremely short and could not be determined. Based on the plasma mAb concentrations in several macaques 24 h following the administration of 20 mg/kg of humanized neutralizing mAbs (viz. approximately 250 μg/ml [Table 13]), the two monkeys receiving 20 mg/kg of 45-46m2 had plasma mAb concentrations of only 15.0 and 17.6 μg/ml, a decay of more than 95% relative to other neutralizing mAbs in 24 h. 
     Neutralization titers were measured on plasma samples collected 24 h following mAb administration when the macaques were challenged with SHIVAD8EO or SHIVDH12-V3AD8. As shown in Table 13, good correlation was observed between anti-viral plasma neutralization titers and protection from SHIV infection. The administration of the two glycan-dependent mAbs (PGT121 and 10-1074) clearly resulted in the highest titers of anti-HIV-1 neutralizing activity at the time of virus challenge. The titers measured in recipients of the 45-46m2 mAb were at the limits of detection or undetectable due to its extremely short half-life in vivo. 
     The method described by Reed and Muench (Am J Hyg 27, 493-497 (1938)) was used to calculate the neutralization titers, measured in plasma, needed to prevent virus acquisition in 50% of challenged monkeys. These protective titers for the 28 monkeys, challenged with SHIVAD8EO, or the 32 monkeys, challenged with SHIVDH12-V3AD8, were separately deduced (Tables 15 and 16). The plasma neutralization titers required for protecting 50% of the SHIVAD8EO or SHIVDH12-V3AD8 challenged animals were calculated to be 1:115 and 1:96, respectively. Because these similar titers were obtained following: 1) SHIV challenges by identical routes and inoculum size and 2) the administration of the same ensemble of neutralizing mAbs, the neutralization data from all 60 animals were combined and subjected to probit regression to examine the relationship between plasma neutralization titers and in vivo protection. As a further check, when a term for the SHIV virus was included in the probit regression model on all 60 macaques, there was no evidence of a difference between the two SHIV viruses (p=0.16). When applied to the entire group of 60 macaques, probit regression estimated that plasma neutralization titers of 1:104 would prevent virus acquisition in 50% of animals. Probit analysis of the data also estimates that 50% plasma neutralization titers of 1:57 or 1:329 would protect 33% or 80%, respectively, of exposed animals. 
     Example 10 Administration of Neutralizing mAbs to Chronically Infected HIV In-Vivo Models 
     Methods Summary: The neutralization activities of the broadly acting 3BNC11724 and 10-107423 neutralizing mAbs against SHIVAD8EO were initially determined in the TZM-bl cell system against SHIVAD8EO. Their capacities to block virus acquisition or to control plasma viremia in chronically infected animals challenged with the R5-tropic SHIVAD8EO were assessed by monitoring plasma viral loads and cell-associated viral nucleic acids; levels of CD4+ cell subsets were measured by flow cytometry. SGA analyses of circulating viral variants and the determination of antibody levels in plasma. Plasma concentration of NAbs was determined by measuring neutralizing activity against HIV-1 pseudovirus preparations only susceptible to either 10-1074 or 3BNC117. 
     Results: 
     Two groups of chronically infected macaques were assessed. The first group consisted of two clinically asymptomatic animals (DBZ3 and DC99A) that had been infected for 159 weeks and had sustained similar and significant declines of circulating CD4+ T cells (Table 17). The regimen for treating ongoing SHIV infections was to co-administer 101074 and 3BNC117, at a dose of 10 mg/kg. At the time of mAb administration, the plasma viral loads in macaques DBZ3 and DC99A were 1.08×104 and 7.6×103 RNA copies/ml, respectively. Both monkeys responded to combination anti-HIV-1 mAb treatment with immediate and rapid reductions of plasma viremia to undetectable levels within 7 to 10 days. Suppression of measurable SHIVAD8EO in the plasma of macaques DBZ3 and DC99A, following a single administration of the two mAbs, lasted 27 and 41 days, respectively. In each case, plasma viremia rebounded to pretreatment levels. 
     A second group of three animals (DBX3, DCF1, and DCM8), each of which were also infected with SHIVAD8EO for more than 3 years and were clinically symptomatic with intermittent diarrhea and or anorexia, were treated with the two neutralizing antibodies (Table 17). At the time of mAb administration, the level of circulating CD4+ T cells in macaque DCM8 was only 43 cells/μ1 and somewhat higher in animals DCF1 (105 cells/μ1) and DBXE (158 cells/μ1). Plasma viral loads exceeded 105 RNA copies/ml in animals DBXE and DCF1 and were significantly lower (1.59×103 RNA copies/ml) in monkey DCM8. The administration of the two mAbs to monkey DBXE resulted in a biphasic reduction of viremia from 2.0×105 RNA copies at day 0 to undetectable levels in plasma at day 20. This was followed, within a few days, by a resurgence of high levels of circulating virus in DBXE. Macaque DCM8, with more modest plasma virus loads and very low numbers of circulating CD4+ T cells, experienced a rapid decline of viremia to undetectable levels between days 6 and 20 following the initiation of mAb treatment. Finally, animal DCF1, previously reported to have generated broadly reacting anti-HIV-1 NAbs, exhibited a transient and a comparatively modest 27-fold reduction of plasma viremia by day 6 in response to combination mAb therapy, before the viral loads returned to high pretreatment levels. 
     PBMC associated viral RNA and DNA levels were also determined prior to and following antibody administration (Table 18). For each animal, mAb treatment resulted in reduced levels of cell associated viral RNA, correlating well with the plasma viral load measurements. No consistent pattern was observed for cell associated viral DNA levels as a result of antibody treatment. Administration of neutralizing mAbs to chronically SHIVAD8EO infected monkeys also had beneficial effects on circulating CD4+ T cell levels, particularly in animals with very high virus loads. The CD4+ T cell numbers in macaques DBXE and DCF1 increased 2 to 3 fold during the period of mAb mediated virus suppression, but gradually declined to pretreatment levels as viremia again became detectable. 
     Plasma concentrations of each mAb were determined by measuring the plasma neutralizing activity against selected HIV-1 pseudovirus strains sensitive to one or the other, but not to both antibodies ( FIG.  13 A ). In every treated animal, suppression of SHIVAD8EO viremia was maintained until a threshold plasma mAb concentration of approximately 1 to 3 μg/ml was reached ( FIGS.  13 B and  13 C ). This was even the case for macaque DCF1, for which a modest and transient reduction of plasma viral RNA levels was observed. Interestingly, the mAbs administered to clinically symptomatic macaques DCM8 and DCF1 had shortened half-lives or were undetectable. As noted earlier, macaque DCM8 had extremely low CD4+ T cell levels (43 cells/μ1 plasma) and macaque DCF1 had to be euthanized on day 56 post treatment initiation due to its deteriorating clinical condition. A necropsy of DCF1 revealed severe enteropathy, characterized by disseminated gastrointestinal cryptosporidiosis, pancreatitis, and cholangitis. 
     SGA analysis was used to determine whether amino acid substitutions had arisen in gp120 regions previously shown to affect the sensitivity to 10-1074 or 3BNC117 mAbs. In each case the rebound virus present in plasma following immunotherapy was unchanged. To further test the sensitivity of the re-emerging viruses, 10-1074 plus 3BNC117 combination therapy (10 mg/kg of each) was re-administered to the two clinically asymptomatic monkeys (DBZ3 and DC99A). The viral loads in each animal again rapidly fell, becoming undetectable at day 7 of the second immunotherapy cycle. Viremia was suppressed for 7 days in macaque DBZ3 and more than 21 days in monkey DC99A. Taken together, these results suggest that the re-emergence of virus following the first treatment cycle in these two animals represented insufficient mAb levels in vivo rather than antibody selected virus resistance. 
     
       
         
           
               
             
               
                 TABLE 3  
               
             
            
               
                   
               
               
                 Repertoire of PGT121 and 10-1074 clonal variants 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 pt10 mAb# 
                 VH 
                 DH 
                 JH 
                 CD43 1   
                 VHmut 
                 Lenght 1   
                 (−) 
                 (+ 
                 Y 
                 Lc 
                 Vλ 
                 Jλ 
                 LCDE3 1   
                 Vλmut 
                 FRW1_del 
                 FRW3_ins 
                 Lenght 1   
                 (−) 
                 (+) 
                 Y 
               
               
                   
               
               
                 10-160 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                 10-186 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 52 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 47 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                 10-248 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 46 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                  10-295* 
                 4-59 
                 / 
                 6 
                 TKHGRRIYGIVAFNEWFTYFYMDV 
                 63 
                 24 
                 2 
                 4 
                 3 
                 λ 
                 3-21 
                 3 
                 HIYDARGGTNWV 
                 58 
                 21 
                 3 
                 12 
                 1 
                 2 
                 1 
               
               
                   
               
               
                 10-266 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 46 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                 10-267 
                 4-59 
                 / 
                 6 
                 AQQGKRIYGIVSFGELFYYYMDA 
                 58 
                 24 
                 2 
                 2 
                 5 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                  10-303* 
                 4-59 
                 3-3/9 
                 6 
                 TLHGRRIYGIVAFNEWFTYFYMDV 
                 54 
                 24 
                 2 
                 3 
                 3 
                 λ 
                 3-21 
                 3 
                 HIWDSRVPTKWV 
                 50 
                 21 
                 3 
                 12 
                 1 
                 3 
                 0 
               
               
                   
               
               
                 10-354 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGWSFGEFFYYYSMDV 
                 48 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                   
                   
               
               
                   
               
               
                  10-410* 
                 4-59 
                 3-10/3 
                 6 
                 ALHGKRIYGIVALGELFTYFYMDV 
                 63 
                 24 
                 2 
                 3 
                 3 
                 λ 
                 3-21 
                 3 
                 HIWDSRRTNWV 
                 44 
                 21 
                 3 
                 12 
                 1 
                 33 
                 0 
               
               
                   
               
               
                 10-416 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 47 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 45 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                 10-468 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 47 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 46 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                 10-543 
                 4-59 
                 3-10/3 
                 6 
                 ALHGKRIYGIVALGELFTYFYMDV 
                 60 
                 24 
                 2 
                 3 
                 3 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                 10-570 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 42 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                 10-621 
                 4-59 
                 3-3/9 
                 6 
                 TLHGRRIYGIVAFNEWFTYFYMDV 
                 54 
                 24 
                 2 
                 3 
                 3 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                 10-664 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 47 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                 10-720 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                 10-730 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 48 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                 10-814 
                 4-59 
                 3-10 
                 6 
                 TQQGKRIYGVVSFGEFFHYYYMDA 
                 43 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HKWDSRSPLSWV 
                 52 
                 15 
                 3 
                 12 
                 1 
                 3 
                 0 
               
               
                   
               
               
                  10-847* 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 47 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 46 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                 10-948 
                 4-59 
                 3-3/6 
                 6 
                 TLHGRRIYGIVAFNEWFTYFYMDV 
                 55 
                 24 
                 2 
                 3 
                 3 
                 λ 
                 3-21 
                 3 
                 HIWDSRVPTKWV 
                 46 
                 21 
                 3 
                 12 
                 1 
                 3 
                 0 
               
               
                   
               
               
                 10-996 
                 4-59 
                 3-3/10 
                 6 
                 TQQGKRIYGVVSFGEFFHYYYMDA 
                 41 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HKWDSRSPLSWV 
                 50 
                 15 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                  10-1022 
                 4-59 
                 3-3 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 51 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                  10-1059 
                 4-59 
                 3-3/16 
                 6 
                 TKHGRRIYGVVAFNEWFTYFYMDV 
                 59 
                 24 
                 2 
                 4 
                 3 
                 λ 
                 3-21 
                 3 
                 HIYDARRPTNWV 
                 46 
                 21 
                 3 
                 12 
                 1 
                 3 
                 1 
               
               
                   
               
               
                   10-1074* 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 45 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                   10-1121* 
                 4-59 
                 3-10/3-3 
                 6 
                 ALHGKRIYGIVALGELFTYFYMDV 
                 63 
                 24 
                 2 
                 3 
                 3 
                 λ 
                 3-21 
                 3 
                 HIWDSRRPTNWV 
                 44 
                 21 
                 3 
                 12 
                 1 
                 3 
                 0 
               
               
                   
               
               
                   10-1130* 
                 4-59 
                 3-10/3-3 
                 6 
                 ALHGKRIYGIVALGELFTYFYMDV 
                 60 
                 24 
                 2 
                 3 
                 3 
                 λ 
                 3-21 
                 3 
                 HIWDSRRPTNWV 
                 42 
                 21 
                 3 
                 12 
                 1 
                 3 
                 0 
               
               
                   
               
               
                   10-1141* 
                 4-59 
                 3-10 
                 6 
                 ALHGKRIYGIVALGELFTYFYMDV 
                 63 
                 24 
                 2 
                 3 
                 3 
                 λ 
                 3-21 
                 3 
                 HIWDSRRPTNWV 
                 45 
                 21 
                 3 
                 12 
                 1 
                 3 
                 0 
               
               
                   
               
               
                   10-1146* 
                 4-59 
                 3-3 
                 6 
                 AQQGKRIYGIVSFGELFYYYMDA 
                 58 
                 24 
                 2 
                 2 
                 5 
                 λ 
                 3-21 
                 3 
                 HYWDSRSPISWV 
                 61 
                 15 
                 3 
                 12 
                 1 
                 2 
                 1 
               
               
                   
               
               
                  10-1151 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 48 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 46 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                  10-1167 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 3 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                  10-1223 
                 4-59 
                 3-10/3-3 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 47 
                 24 
                 2 
                 3 
                 4 
                 / 
                 / 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                  10-1232 
                 4-59 
                 3-3/16 
                 6 
                 ALHGKRIYGIVALGELFTYFYMDV 
                 58 
                 24 
                 2 
                 3 
                 3 
                 / 
                 3-21 
                 / 
                 / 
                 / 
                   
                   
                 / 
                 / 
                 / 
                 / 
               
               
                   
               
               
                  10-1263 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 46 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                  10-1294 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 45 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                   10-1341* 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 49 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 45 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                  10-1342 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 48 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 46 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                   10-1369* 
                 4-59 
                 3-3/16 
                 6 
                 TKHGRRIYGVVAFGEWFTYFYMDV 
                 57 
                 24 
                 2 
                 4 
                 3 
                 λ 
                 3-21 
                 3 
                 HIYDARRPTNWV 
                 43 
                 21 
                 3 
                 12 
                 1 
                 3 
                 1 
               
               
                   
               
               
                  10-1476 
                 4-59 
                 3-3/16 
                 6 
                 ARRGQRIYGVVSFGEFFYYYSMDV 
                 47 
                 24 
                 2 
                 3 
                 4 
                 λ 
                 3-21 
                 3 
                 HMWDSRSGFSWS 
                 46 
                 12 
                 3 
                 12 
                 1 
                 2 
                 0 
               
               
                   
               
               
                 VHmut and Vλmut indicate the total number of mutations in the VH and VL lg genes. 
               
               
                 (−) and (+) indicate the number of negatively and positively charged ammino acids in the lg complementary determining region (CDR3), respectively. 
               
               
                 Y indicated the number of Tyrosine residue in the lgH/L CDRs. 
               
               
                   1 Based on Kabat nomenclature (lgBLAST). 
               
               
                 FRW1_del, number of deleted nucleotides in framework region 1 (FRW1) of the lgL. 
               
               
                 FRW3_ins, number of inserted nucleotides in framework region 3 (FRW3) of the lgL. 
               
               
                 Clonal members with identical lgH sequences are indicated and among them, lgL sequence identity that defines clones. 
               
               
                 *indicates the representative antibody variants that were produced and analyzed. 
               
               
                 10-266 lgL was not cloned, and 10-1141 lgG was not produced. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
               
                   
               
               
                 In vitro TZM-bl neutralization assay on the basic pane 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 10-1369 
                 10-259 
                 PGT121 
                 10-303 
                 10-410 
                 10-1130 
               
               
                   
               
               
                 IC50 
                   
                   
                   
                   
                   
                   
               
               
                 BaL.26 
                 0.069 
                 0.021 
                 0.021 
                 0.045 
                 0.016 
                 0.013 
               
               
                 SS1196.1 
                 0.033 
                 0.012 
                 0.008 
                 0.015 
                 0.008 
                 0.008 
               
               
                 6535.3 
                 0.023 
                 0.005 
                 0.007 
                 0.014 
                 0.003 
                 0.003 
               
               
                 QH0692.42 
                 0.503 
                 0.155 
                 1.085 
                 3.122 
                 2.830 
                 4.871 
               
               
                 TRJO4551.58 
                 0.569 
                 0.189 
                 3.896 
                 14.401 
                 18.511 
                 36.880 
               
               
                 SC422661.8 
                 0.195 
                 0.096 
                 0.263 
                 0.333 
                 0.132 
                 0.070 
               
               
                 PVO.4 
                 0.225 
                 0.175 
                 0.147 
                 0.670 
                 0.494 
                 0.385 
               
               
                 CAAN5342.A2 
                 0.070 
                 0.020 
                 0.013 
                 0.020 
                 0.012 
                 0.009 
               
               
                 YU-2 
                 0.210 
                 0.135 
                 0.098 
                 0.190 
                 0.089 
                 0.078 
               
               
                 R1166.c1 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                 MuLV 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                 IC80 
                   
                   
                   
                   
                   
                   
               
               
                 Bal.26 
                 0.268 
                 0.101 
                 0.081 
                 0.156 
                 0.066 
                 0.062 
               
               
                 SS1196.1 
                 0.033 
                 0.037 
                 0.030 
                 0.055 
                 0.030 
                 0.037 
               
               
                 6535.3 
                 0.060 
                 0.022 
                 0.041 
                 0.053 
                 0.021 
                 0.013 
               
               
                 QH0692.42 
                 1.714 
                 0.551 
                 14.976 
                 18.122 
                 12.071 
                 &gt;40 
               
               
                 TRJO4551.58 
                 3.818 
                 0.965 
                 26.930 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                 SC422661.8 
                 0.940 
                 0.333 
                 0.714 
                 1.156 
                 0.449 
                 0.264 
               
               
                 PVO.4 
                 0.787 
                 0.716 
                 1.097 
                 2.199 
                 1.572 
                 1.783 
               
               
                 CAAN5342.A2 
                 0.186 
                 0.063 
                 0.056 
                 0.092 
                 0.055 
                 0.045 
               
               
                 YU-2 
                 0.738 
                 0.382 
                 0.356 
                 0.502 
                 0.243 
                 0.313 
               
               
                 R1166.c1 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                 MuLV 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                   
               
               
                   
                 10-1121 
                 10-1146 
                 10-996 
                 10-1341 
                 10-847 
                 10-1074 
               
               
                   
               
               
                 IC50 
                   
                   
                   
                   
                   
                   
               
               
                 BaL.26 
                 0.046 
                 0.064 
                 0.045 
                 0.032 
                 0.022 
                 0.033 
               
               
                 SS1196.1 
                 0.029 
                 0.027 
                 0.007 
                 0.011 
                 0.006 
                 0.010 
               
               
                 6535.3 
                 0.008 
                 0.022 
                 0.018 
                 0.009 
                 0.011 
                 0.007 
               
               
                 QH0692.42 
                 4.187 
                 0.590 
                 0.395 
                 0.335 
                 0.259 
                 0.259 
               
               
                 TRJO4551.58 
                 15.360 
                 0.548 
                 0.516 
                 0.333 
                 0.210 
                 0.170 
               
               
                 SC422661.8 
                 0.173 
                 0.195 
                 0.255 
                 0.189 
                 0.137 
                 0.145 
               
               
                 PVO.4 
                 0.570 
                 0.310 
                 0.211 
                 0.236 
                 0.172 
                 0.178 
               
               
                 CAAN5342.A2 
                 0.033 
                 0.032 
                 0.007 
                 0.009 
                 0.006 
                 0.007 
               
               
                 YU-2 
                 0.152 
                 0.275 
                 0.256 
                 0.234 
                 0.161 
                 0:143 
               
               
                 R1166.c1 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                 MuLV 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                 IC80 
                   
                   
                   
                   
                   
                   
               
               
                 Bal.26 
                 0.154 
                 0.203 
                 0.228 
                 0.159 
                 0.112 
                 0.124 
               
               
                 SS1196.1 
                 0.098 
                 0.073 
                 0.040 
                 0.040 
                 0.026 
                 0.127 
               
               
                 6535.3 
                 0.033 
                 0.078 
                 0.085 
                 0.038 
                 0.044 
                 0.044 
               
               
                 QH0692.42 
                 21.943 
                 1.993 
                 1.404 
                 1.100 
                 0.908 
                 0.861 
               
               
                 TRJO4551.58 
                 &gt;23 
                 2.604 
                 4.265 
                 1.226 
                 0.768 
                 0.693 
               
               
                 SC422661.8 
                 0.741 
                 0.663 
                 0.845 
                 0.501 
                 0.386 
                 0.392 
               
               
                 PVO.4 
                 2.465 
                 1.319 
                 1.715 
                 0.754 
                 0.774 
                 0.766 
               
               
                 CAAN5342.A2 
                 0.095 
                 0.088 
                 0.060 
                 0.054 
                 0.035 
                 0.044 
               
               
                 YU-2 
                 0.340 
                 0.750 
                 0.891 
                 0.766 
                 0.537 
                 0.398 
               
               
                 R1166.c1 
                 &gt;23 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                 MuLV 
                 &gt;23 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
                 &gt;40 
               
               
                   
               
               
                 Numbers indicate antibody lgG concentrations in μg/ml to reach the IC 50  (top) and IC 80  (bottom) in the TZM-bl neutralization assay. 
               
               
                 IC 50/80  values are indicated. 
               
               
                 &gt;indicates that the IC 50  for a given virus was not reached at the concentration tested. 
               
               
                 Murine leukemia virus (MuLV) and R1166.c1 (clade AE) are negative controls. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 In vitro TZM-bl neutralization assay on the extended panel - IC50 values 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 Virus ID 
                 Clade 
                 10-996 
                 10-1074 
                 PGT121 
                 Virus ID 
                 Clade 
                 10-996 
                 10-1074 
                 PGT121 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 6535.3 
                 B 
                 0.017 
                 0.014 
                 0.008 
                 CNE58 
                 BC 
                 0.570 
                 0.267 
                 &gt;50 
               
               
                 QH0692.42 
                 B 
                 0.396 
                 0.191 
                 1.041 
                 MS208.A1 
                 A 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 SC422661.8 
                 B 
                 0.173 
                 0.091 
                 0.101 
                 Q23.17 
                 A 
                 0.008 
                 0.006 
                 0.010 
               
               
                 PVO.4 
                 B 
                 0.186 
                 0.074 
                 0.131 
                 Q451.e2 
                 A 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 TR0.11 
                 B 
                 0.012 
                 0.008 
                 0.005 
                 Q769.d22 
                 A 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 AC10.0.29 
                 B 
                 0.067 
                 0.022 
                 0.037 
                 Q259.c12.17 
                 A 
                 &gt;50 
                 &gt;50 
                 8.990 
               
               
                 RHPA4259.7 
                 B 
                 0.034 
                 0.021 
                 0.014 
                 Q842.d.12 
                 A 
                 &gt;50 
                 &gt;50 
                 0.023 
               
               
                 THRO4156.18 
                 B 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 3415.v1.c1 
                 A 
                 35.876 
                 &gt;50 
                 &gt;50 
               
               
                 REJO4541.67 
                 B 
                 &gt;50 
                 &gt;50 
                 3.607 
                 3365.v2.c2 
                 A 
                 0.286 
                 0.131 
                 0.921 
               
               
                 TRJO4551.58 
                 B 
                 0.147 
                 0.170 
                 3.728 
                 0260.v5.c36 
                 A 
                 0.160 
                 0.099 
                 0.054 
               
               
                 WITO4160.33 
                 B 
                 0.538 
                 0.185 
                 0.459 
                 191955_A11 
                 A(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 CAAN5342.A2 
                 B 
                 0.013 
                 0.007 
                 0.011 
                 191084 B7-19 
                 A(T/F) 
                 0.057 
                 0.032 
                 0.042 
               
               
                 YU-2 
                 B 
                 0.256 
                 0.143 
                 0.098 
                 9004SS_A3_4 
                 A(T/F) 
                 0.012 
                 0.011 
                 0.008 
               
               
                 WEAU_d15_410_787 
                 B(T/F) 
                 0.147 
                 0.104 
                 0.083 
                 T257-31 
                 CRF02_AG 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 1006_11_C3_1601 
                 B(T/F) 
                 0.001 
                 0.003 
                 0.008 
                 928-28 
                 CRF02_ AG 
                 1.331 
                 0.847 
                 &gt;50 
               
               
                 1054_07_TC4_1499 
                 B(T/F) 
                 0.260 
                 0.129 
                 0.115 
                 263-8 
                 CRF02_AG 
                 10.919 
                 0.666 
                 3.347 
               
               
                 1056_10_TA11_1826 
                 B(T/F) 
                 0.117 
                 0.038 
                 0.066 
                 T250-4 
                 CRF02_AG 
                 &lt;0.001 
                 &lt;0.001 
                 0.001 
               
               
                 1012_11_TC21_3257 
                 B(T/F) 
                 0.018 
                 0.008 
                 0.008 
                 T251-18 
                 CRF02_AG 
                 0.939 
                 1.081 
                 &gt;50 
               
               
                 6240_08_TA5_4622 
                 B(T/F) 
                 0.095 
                 0.068 
                 0.128 
                 T278-50 
                 CRF02_AG 
                 14.010 
                 2.146 
                 &gt;50 
               
               
                 6244_13_85_4576 
                 B(T/F) 
                 0.353 
                 0.202 
                 0.249 
                 T255-34 
                 CRF02_AG 
                 28.369 
                 &gt;50 
                 6.725 
               
               
                 62357_14_D3_4589 
                 B(T/F) 
                 29.300 
                 &gt;50 
                 1.036 
                 211-9 
                 CRF02_AG 
                 0.750 
                 0.112 
                 1.455 
               
               
                 SC05_8C11_2344 
                 B(T/F) 
                 0.069 
                 0.052 
                 0.093 
                 235-47 
                 CRF02_AG 
                 0.128 
                 0.050 
                 0.332 
               
               
                 Du156.12 
                 C 
                 0.018 
                 0.015 
                 0.007 
                 620345.c01 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 Du172.17 
                 C 
                 0.173 
                 0.121 
                 0.115 
                 CNE8 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 Du422.1 
                 C 
                 0.056 
                 0.045 
                 0.029 
                 C1080.c03 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 ZM197M.PB7 
                 C 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 R2184.c04 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 ZM2145M.PL15 
                 C 
                 0.413 
                 0.174 
                 0.236 
                 R1166.c01 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 ZM233M.PB6 
                 C 
                   
                 0.060 
                 1.451 
                 C2101.c01 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 ZM249M.PL1 
                 C 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 C3347.c11 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 ZM53M.PB12 
                 C 
                 &gt;50 
                 &gt;50 
                 &lt;0.001 
                 C4118.c09 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 ZM109F.PB4 
                 C 
                 &gt;50 
                 &gt;50 
                 7.894 
                 CNE5 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 ZM135M.PL10a 
                 C 
                 0.099 
                 0.069 
                 0.576 
                 BJOX009000.02.4 
                 CRF01_AE 
                 &gt;50 
                 &gt;50 
                 3.626 
               
               
                 CAP45.2.00.G3 
                 C 
                 &gt;50 
                 &gt;50 
                 0.086 
                 BJOX015000.11.5 
                 CRF01_AE(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 CAP210.2.00.E8 
                 C 
                 24.793 
                 &gt;50 
                 5.082 
                 BJOX010000.06.2 
                 CRF01_AE(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 HIV-001428-2.42 
                 C 
                 0.040 
                 0.044 
                 0.028 
                 BJOX025000.01.1 
                 CRF01_AE(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 HIV-0013095-2.11 
                 C 
                 31.531 
                 &gt;50 
                 &gt;50 
                 BJOX028000.10.3 
                 CRF01_AE(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 HIV-16055-2.3 
                 C 
                 &gt;50 
                 &gt;50 
                 0.444 
                 X1193_c1 
                 G 
                 0.144 
                 0.083 
                 0.045 
               
               
                 H3V-16845-2.22 
                 C 
                 1.325 
                 1.169 
                 12.685 
                 P0402_c2_11 
                 G 
                 0.022 
                 0.012 
                 0.020 
               
               
                 Ce1086_B2 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &lt;0.001 
                 X1254_c3 
                 G 
                 0.121 
                 0.089 
                 0.056 
               
               
                 Ce0393_C3 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 X2088_c9 
                 G 
                 0.002 
                 0.003 
                 0.011 
               
               
                 Ce1176_A3 
                 C(T/F) 
                 0.043 
                 0.018 
                 0.017 
                 X2131_C1_B5 
                 G 
                 0.019 
                 0.016 
                 0.015 
               
               
                 Ce2010_F5 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 P1981_C5_3 
                 G 
                 0.005 
                 0.005 
                 0.004 
               
               
                 Ce0682_E4 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 X1632_S2_B10 
                 G 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 C1172_H1 
                 C(T/F) 
                 0.058 
                 0.047 
                 0.023 
                 30.16.v5.c45 
                 D 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 Ce2060_G9 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 A07412M1.vrc12 
                 D 
                 0.008 
                 &lt;0.001 
                 0.001 
               
               
                 Ce703010054_2A2 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 231965.c01 
                 D 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 8F1266.431a 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 231966.c02 
                 D 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 246F C1G 
                 C(T/F) 
                 0.092 
                 0.022 
                 0.083 
                 191821_E6_1 
                 D(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 249M B10 
                 C(T/F) 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 3817.v2.c59 
                 CD 
                 8.147 
                 3.148 
                 &gt;50 
               
               
                 ZM247v1(Rev-) 
                 C(T/F) 
                 0.055 
                 0.042 
                 0.027 
                 6480.v4.c25 
                 CD 
                 0.010 
                 0.009 
                 0.017 
               
               
                 7030102001E5(Rev-) 
                 C(T/F) 
                 0.013 
                 0.006 
                 0.010 
                 6952.v1.c20 
                 CD 
                 0.044 
                 0.037 
                 0.085 
               
               
                 1394C9G1(Rev-) 
                 C(T/F) 
                 0.086 
                 0.050 
                 0.486 
                 6811.v7.c18 
                 CD 
                 0.001 
                 0.002 
                 0.004 
               
               
                 Ce704809221_1B3 
                 C(T/F) 
                 0.243 
                 0.139 
                 0.098 
                 89-F1_2_25 
                 CD 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 CNE19 
                 BC 
                 3.452 
                 50.000 
                 0.018 
                 3301.v1.c24 
                 AC 
                 0.016 
                 0.013 
                 0.014 
               
               
                 CNE20 
                 BC 
                 &lt;0.001 
                 &lt;0.001 
                 0.002 
                 6041.v3_c23 
                 AC 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 CNE21 
                 BC 
                 0.086 
                 0.087 
                 0.020 
                 6540.v4.c1 
                 AC 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 CNE17 
                 BC 
                 4.040 
                 2.686 
                 45.289 
                 6545.v4.c1 
                 AC 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 CNE30 
                 BC 
                 0.614 
                 0.363 
                 0.101 
                 0815.v3.c3 
                 ACD 
                 0.061 
                 0.030 
                 0.022 
               
               
                 CNE52 
                 BC 
                 4.525 
                 1.226 
                 3.741 
                 3103.v3.010 
                 ACD 
                 0.053 
                 0.037 
                 0.042 
               
               
                 CNE53 
                 BC 
                 0.057 
                 0.039 
                 0.055 
               
               
                   
               
               
                 Numbers indicate antibody lgG concentrations in μg/ml to reach the IC 50  in the TZM-bl neutralization assay. 
               
               
                 IC50 values indicate increasing neutralization sensitivity. 
               
               
                 &gt;indicates that the IC 50  for a given virus was not reached at the concentration tested. 
               
            
           
         
       
     
                     TABLE 6                  In vitro TZM-bl neutralization assay on the extended panel - IC80                                                     Virus ID   Clade   10-996   10-1074   PGT121   Virus ID   Clade   10-996   10-1074   PGT121                                                             6535.3   B   0.046   0.026   0.021   CNE58   BC   2.220   0.968   &gt;50       QH0692.42   B   1.854   0.929   8.545   MS208.A1   A   &gt;50   &gt;50   &gt;50       SC422661.8   B   0.627   0.418   0.460   Q23.17   A   0.030   0.021   0.031       PVO.4   B   0.952   0.360   0.945   Q451.e2   A   &gt;50   &gt;50   &gt;50       TR0.11   B   0.081   0.057   0.051   Q769.d22   A   &gt;50   &gt;50   &gt;50       AC10.0.29   B   0.250   0.110   0.169   Q259.d2.17   A   &gt;50   &gt;50   &gt;50       RHPA4259.7   B   0.163   0.113   0.054   Q342.d12   A   &gt;50   &gt;50   0.074       THRO4156.18   B   &gt;50   &gt;50   &gt;50   3415.v1.01   A   &gt;50   &gt;50   &gt;50       REJO4541.67   B   &gt;50   &gt;50   &gt;50   3365.v2.02   A   1.380   0.450   7.353       TRJO4551.58   B   7.269   0.634   35.291   0260.v5.c36   A   0.436   0.160   0.152       WITO4160.33   B   6.484   2.112   6.007   191955_A11   A(T/F)   &gt;50   &gt;50   &gt;50       CAAN5342.A2   B   0.079   0.036   0.051   191084 B7-19   A(T/F)   0.144   0.128   0.128       YU-2   B   0.891   0.398   0.356   9004SS_A3_4   A(T/F)   0.050   0.030   0.026       WEAU_d15_410_787   B(T/F)   0.422   0.375   0.295   T257-31   CRF02_AG   &gt;50   &gt;50   &gt;50       1006_11_C3_1601   B(T/F)   0.019   0.013   0.023   928-28   CRF02_AG   7.151   4.696   &gt;50       1054_07_TC4_1499   B(T/F)   0.901   0.563   0.696   263-8   CRF02_AG   &gt;50   6.527   24.576       1054_10_TA11_1826   B(T/F)   0.563   0.272   0.303   T250-4   CRF02_AG   0.005   0.005   0.011       1012_11_TC21_3257   B(T/F)   0.111   0.059   0.038   T251-18   CRF02_AG   7.399   7.395   &gt;50       6240_08_TA5_4622   B(T/F)   0.348   0.306   0.584   T278-50   CRF02_AG   &gt;50   18.276   &gt;50       6244_13_B5_4576   B(T/F)   1.296   0.922   1.878   T255-34   CRF02_AG   &gt;50   &gt;50   &gt;50       62357_14_D3_4589   B(T/F)   &gt;50   &gt;50   45.559   211-9   CRF02_AG   3.848   0.425   8.840       SC05_8C11_2344   B(T/F)   0.174   0.123   0.275   235-47   CRF02_AG   0.381   0.163   1.676       Du156.12   C   0.101   0.076   0.033   620345.c01   CRF01_AE   &gt;50   &gt;50   &gt;50       Du172.17   C   0.607   0.430   0.890   CNE8   CRF01_AE   &gt;50   &gt;50   &gt;50       Du422.1   C   0.215   0.166   0.131   C1080.c03   CRF01_AE   &gt;50   &gt;50   &gt;50       ZM197M.PB7   C   &gt;50   &gt;50   &gt;50   R2184.c04   CRF01_AE   &gt;50   &gt;50   &gt;50       ZM214M.PL15   C   3.251   2.367   3.150   R1166.c01   CRF01_AE   &gt;50   &gt;50   &gt;50       ZM233M.PB6   C   4.524   0.349   8.977   C2101.c01   CRF01_AE   &gt;50   &gt;50   &gt;50       ZM249M.PL1   C   &gt;50   &gt;50   &gt;50   C3347.c11   CRF01_AE   &gt;50   &gt;50   &gt;50       ZM53M.PB12   C   &gt;50   &gt;50   0.002   C4118.009   CRF01_AE   &gt;50   &gt;50   &gt;50       ZM109F.PB4   C   &gt;50   &gt;50   &gt;50   CNE5   CRF01_AE   &gt;50   &gt;50   &gt;50       ZM135M.PL10a   C   0.553   0.367   5.885   BJOX009000.02.4   CRF01_AE   &gt;50   &gt;50   37.289       CAP45.2.00.G3   C   &gt;50   &gt;50   6.544   BJOX015000.11.5   CRF01_AE(T/F)   &gt;50   &gt;50   &gt;50       CAP210.2.00.E8   C   &gt;50   &gt;50   &gt;50   BJOX010000.06.2   CRF01_AE(T/F)   &gt;50   &gt;50   &gt;50       HIV-001428-2.42   C   0.204   0.261   0.156   BJOX025000.01.1   CRF01_AE(T/F)   &gt;50   &gt;50   &gt;50       HIV-0013095-2.11   C   &gt;50   &gt;50   &gt;50   BJOX028000.10.3   CRF01_AE(T/F)   &gt;50   &gt;50   &gt;50       H1V-16055-2.3   C   &gt;50   &gt;50   4.290   X1193_c1   G   0.482   0.475   0.202       H1V-16845-2.22   C   9.933   5.835   &gt;50   P0402_c2_11   G   0.065   0.039   0.056       Ce1086_B2   C(T/F)   &gt;50   &gt;50   0.006   X1254_c3   G   0.420   0.297   0.199       Ce0393_C3   C(T/F)   &gt;50   &gt;50   &gt;50   X2088_c9   G   0.014   0.014   0.029       Ce1176_A3   C(T/F)   0.151   0.070   0.058   X2131_C1_B5   G   0.085   0.064   0.058       Ce2010_F5   C(T/F)   &gt;50   &gt;50   &gt;50   P1981_C5_3   G   0.018   0.017   0.015       Ce0682_E4   C(T/F)   &gt;50   &gt;50   &gt;50   X1632_S2_B10   G   &gt;50   &gt;50   &gt;50       Ce1172_H1   C(T/F)   0.173   0.166   0.088   3016.v5.c45   13   &gt;50   &gt;50   &gt;50       Ce2060_G9   C(T/F)   &gt;50   &gt;50   &gt;50   A07412M1.vrc12   0   0.070   0.048   0.406       Ce703010054_2A2   C(T/F)   &gt;50   &gt;50   &gt;50   231965.c01   D   &gt;50   &gt;50   &gt;50       BF1266.431a   C(T/F)   &gt;50   &gt;50   &gt;50   231966.c02   D   &gt;50   &gt;50   &gt;50       246F C1G   C(T/F)   0.270   0.111   0.287   191821_E6_1   D(T/F)   &gt;50   &gt;50   &gt;50       249M B10   C(T/F)   &gt;50   &gt;50   &gt;50   3817.v2.c59   CD   34.619   14.880   &gt;50       ZM247v1(Rev-)   C(T/F)   0.252   0.186   0.126   6480.v4.c25   CD   0.049   0.041   0.079       7030102001E5(Rev-)   C(T/F)   0.044   0.021   0.043   6952.v1.020   CD   0.188   0.138   0.605       1394C9G1(Rev-)   C(T/F)   0.328   0.191   3.372   6811.v7.018   CD   0.011   0.010   0.017       Ce704809221_1B3   C(T/F)   1.208   0.696   0.492   89-F1_2_25   CD   &gt;50   &gt;50   &gt;50       CNE19   BC   &gt;50   &gt;50   0.189   3301.v1.024   AC   0.054   0.042   0.043       CNE20   BC   &lt;0.001   0.005   0.008   6041.v3.c23   AC   &gt;50   &gt;50   &gt;50       CNE21   BC   0.255   0.181   0.061   6540.v4.01   AC   &gt;50   &gt;50   &gt;50       CNE17   BC   24.701   13.297   &gt;50   6545.v4.01   AC   &gt;50   &gt;50   &gt;50       CNE30   BC   1.989   1.200   0.559   0815.v3.c3   ACD   0.251   0.138   0.105       CNE52   BC   43.834   13.147   32.935   3103.v3.c10   ACD   0.150   0.101   0.110       CNE53   BC   0.233   0.141   0.200               Numbers indicate antibody lgG concentrations in μg/ml to reach the IC 80  in the TZM-bl enuralization assay.       IC 80  values indicate neutralization sensitivity.       &gt;indicates that the IC 80  for a given virus was not reached at the concentration tested.            
Table 7 Neutralization sensitivity according to N332 PNGS (shown in  FIG.  14   ).
 
     
       
         
           
               
             
               
                 TABLE 8 
               
               
                   
               
               
                 In vitro PBMC-based neutralization assay 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 Virus ID 
                 3BNC55 
                 3BNC60 
                 3BNC117 
                 3BNC134 
                 1NC9 
                 45-46 
                 3BNC195 
                 12Al2 
                 4E10 
               
               
                   
               
               
                 CONTEM- 
                 P035.6.E4 
                 1.918 
                 0.023 
                 &lt;0.0032 
                 0.034 
                 0.451 
                 0.037 
                 &gt;50 
                 0.110 
                 1.043 
               
               
                 PORARY 
                 P035.6.H11 
                 0.550 
                 0.029 
                 0.022 
                 0.239 
                 0.031 
                 0.053 
                 &gt;50 
                 0.130 
                 2.941 
               
               
                   
                 P035.6.D10 
                 &gt;50 
                 &gt;50 
                 0.019 
                 &gt;50 
                 0.248 
                 &lt;0.0032 
                 &gt;50 
                 &lt;0.0032 
                 0.792 
               
               
                   
                 P151.37.C7 
                 0.084 
                 0.038 
                 0.053 
                 0.10 
                 0.018 
                 0.016 
                 0.133 
                 0.402 
                 2.941 
               
               
                   
                 P151.37.F1 
                 1.297 
                 0.125 
                 0.161 
                 1.60 
                 1.288 
                 0.203 
                 0.136 
                 18.043 
                 0.860 
               
               
                   
                 P151.37.F10 
                 3.770 
                 0.311 
                 0.20 
                 4.661 
                 &gt;50 
                 0.375 
                 &gt;50 
                 18.863 
                 9.314 
               
               
                   
                 P153.10.2.A9 
                 15.804 
                 0.763 
                 0.861 
                 17.208 
                 46.735 
                 0.069 
                 0.201 
                 &gt;50 
                 12.269 
               
               
                   
                 P153:10.2.D8 
                 20.568 
                 1.020 
                 0.206 
                 23.124 
                 &gt;50 
                 0.030 
                 &gt;50 
                 2.478 
                 0.861 
               
               
                   
                 P153.10.2.E10 
                 1.226 
                 0.546 
                 0.211 
                 2.105 
                 &gt;50 
                 0.108 
                 &gt;50 
                 10.686 
                 0.792 
               
               
                   
                 P186.12.1.D10 
                 0.051 
                 0.061 
                 0.105 
                 &gt;50 
                 0.075 
                 0.165 
                 0.091 
                 7.288 
                 &gt;25 
               
               
                   
                 P186.12.1.F4 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 4.903 
               
               
                   
                 P186.21.1.G2 
                 1.890 
                 &lt;0.0032 
                 &lt;0.0032 
                 &gt;50 
                 0.046 
                 0.017 
                 &gt;50 
                 0.068 
                 15.711 
               
               
                   
                 P195.31.A6 
                 0.032 
                 0.138 
                 &lt;0.0032 
                 0.171 
                 0.228 
                 0.052 
                 0.099 
                 0.206 
                 2.934 
               
               
                   
                 P195.31.A10 
                 0.252 
                 0.024 
                 0.025 
                 &gt;50 
                 0.953 
                 0.050 
                 0.071 
                 &gt;50 
                 1.248 
               
               
                   
                 P195.31F11 
                 0.569 
                 0.084 
                 0.137 
                 &gt;50 
                 &gt;50 
                 0.076 
                 &gt;50 
                 33.131 
                 2.301 
               
               
                   
                 P019.1.D2 
                 0.949 
                 0.164 
                 0.027 
                 &gt;50 
                 &gt;50 
                 0.057 
                 &gt;50 
                 14.612 
                 &gt;25 
               
               
                   
                 P019.1.D8 
                 3.122 
                 0.738 
                 0.635 
                 &gt;50 
                 &gt;50 
                 5.767 
                 &gt;50 
                 &gt;50 
                 2.443 
               
               
                   
                 P019.1.G7 
                 6.034 
                 3.636 
                 4.570 
                 &gt;50 
                 &gt;50 
                 6.962 
                 &gt;50 
                 20.092 
                 1.073 
               
               
                   
                 P175.10.D7 
                 2.539 
                 0.936 
                 3.588 
                 &gt;50 
                 42.695 
                 17.723 
                 &gt;50 
                 35.805 
                 1.187 
               
               
                   
                 P175.10.D12 
                 0.708 
                 0.410 
                 0.067 
                 &gt;50 
                 0.317 
                 0.175 
                 1.233 
                 10.648 
                 0.953 
               
               
                   
                 P175.10.G10 
                 2.508 
                 0.563 
                 0.621 
                 &gt;50 
                 33.008 
                 8.364 
                 22.413 
                 18.554 
                 1.147 
               
               
                   
                 P013.18.A9 
                 0.125 
                 0.102 
                 0.079 
                 &gt;50 
                 0.198 
                 0.018 
                 0.426 
                 0.205 
                 0.786 
               
               
                   
                 P154.44.C8 
                 &gt;50 
                 0.683 
                 1.042 
                 &gt;50 
                 &gt;50 
                 16.440 
                 0.925 
                 &gt;50 
                 &gt;25 
               
               
                   
                 P154.44.G8 
                 &gt;50 
                 1.220 
                 2.172 
                 &gt;50 
                 &gt;50 
                 25.616 
                 1.969 
                 &gt;50 
                 4.450 
               
               
                   
                 P183.50.2.H3 
                 2.197 
                 0.116 
                 0.1212 
                 12.049 
                 4.079 
                 0.161 
                 &gt;50 
                 0.476 
                 3.890 
               
               
                   
                 P1833.2.B9 
                 0.076 
                 0.025 
                 &lt;0.0032 
                 0.053 
                 0.326 
                 0.142 
                 &gt;50 
                 0.472 
                 1.722 
               
               
                   
                 P001.35.F5 
                 0.418 
                 0.046 
                 0.047 
                 8.133 
                 0.329 
                 0.133 
                 &gt;50 
                 0.197 
                 &gt;0.39 
               
               
                   
                 P001.35 H4 
                 2.180 
                 0.333 
                 1.913 
                 &gt;50 
                 &gt;50 
                 6.118 
                 &gt;50 
                 &gt;50 
                 0.830 
               
               
                   
                 P002.39.CB 
                 15.016 
                 0.117 
                 0.172 
                 2.192 
                 12.154 
                 0.546 
                 &gt;50 
                 &gt;50 
                 1.955 
               
               
                   
                 P002.39.F8 
                 &gt;50 
                 28.391 
                 &gt;50 
                 13.090 
                 &gt;50 
                 0.404 
                 &gt;50 
                 &gt;50 
                 0.864 
               
               
                   
                 P002.39.H10 
                 1.472 
                 0.785 
                 0.728 
                 14.169 
                 1.147 
                 0.017 
                 &gt;50 
                 &gt;50 
                 1.530 
               
               
                   
                 P034.6.D6 
                 37.003 
                 0.161 
                 0.176 
                 11.339 
                 1.137 
                 0.235 
                 &gt;50 
                 0.802 
                 1.164 
               
               
                   
                 P034.6.G19 
                 48.877 
                 0.348 
                 0.237 
                 22.481 
                 1.428 
                 0.017 
                 &gt;50 
                 0.186 
                 0.820 
               
               
                   
                 P034.6 H5 
                 &gt;50 
                 0.417 
                 0.267 
                 20.730 
                 0.820 
                 0.245 
                 &gt;50 
                 0.866 
                 0.391 
               
               
                   
                 P101.20.1.F1 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 0.565 
                 &gt;50 
                 0.634 
                 3.999 
               
               
                   
                 P101.20.1 HB 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 0.090 
                 &gt;50 
                 0.474 
                 1.902 
               
               
                   
                 P127.46.A6 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 0.385 
                 &gt;50 
                 0.666 
                 1.211 
               
               
                   
                 P127.46.D1 
                 1.242 
                 0.024 
                 0.043 
                 5.403 
                 2.558 
                 0.169 
                 &gt;50 
                 0.227 
                 1.092 
               
               
                   
                 P127.45.D2 
                 1.125 
                 0.173 
                 0.221 
                 &gt;50 
                 2.231 
                 0.279 
                 &gt;50 
                 0.494 
                 1.613 
               
               
                   
                 P174.28.E11 
                 2.399 
                 0.483 
                 0.716 
                 &gt;50 
                 13.061 
                 0.894 
                 &gt;50 
                 2.104 
                 2.113 
               
               
                   
                 P177.25.1.G9 
                 0.980 
                 0.191 
                 0.189 
                 &gt;50 
                 1.826 
                 0.261 
                 &gt;50 
                 0.130 
                 1.665 
               
               
                   
                 P177.25.2.B4 
                 1.179 
                 0.080 
                 0.041 
                 23.609 
                 0.384 
                 0.150 
                 &gt;50 
                 0.028 
                 3.729 
               
               
                   
                 P177.25.2.D1 
                 &gt;50 
                 1.949 
                 1.359 
                 &gt;50 
                 46.825 
                 11.454 
                 &gt;50 
                 7.681 
                 1.140 
               
               
                   
                 P180.14.A6 
                 1.389 
                 0.098 
                 0.058 
                 0.017 
                 &gt;50 
                 0.024 
                 &gt;50 
                 0.022 
                 1.162 
               
               
                   
                 P130.14.G6 
                 45.246 
                 0.116 
                 0.122 
                 2.449 
                 13.361 
                 0.169 
                 0.220 
                 0.093 
                 4.009 
               
               
                   
                 P180.14.G7 
                 23.444 
                 0.052 
                 0.035 
                 0.022 
                 1.450 
                 0.024 
                 0.703 
                 0.729 
                 15.759 
               
               
                   
                 P197.251.D2 
                 &gt;50 
                 0.285 
                 0.194 
                 1.480 
                 1.137 
                 0.016 
                 0.028 
                 0.072 
                 1.492 
               
               
                   
                 P197.25.1.D7 
                 &gt;50 
                 0.782 
                 0.019 
                 0.052 
                 7.056 
                 0.051 
                 0.016 
                 2.601 
                 0.943 
               
               
                   
                 P197.25.1.H1 
                 &gt;50 
                 0.017 
                 0.019 
                 &lt;0 0032 
                 &lt;0.0032 
                 0.029 
                 &lt;0.0032 
                 0.754 
                 2.515 
               
               
                   
                 P405.18.D3 
                 0 068 
                 &lt;0.0032 
                 0.029 
                 0.095 
                 0.048 
                 0.022 
                 0.022 
                 0.031 
                 0.924 
               
               
                   
                 P405.18.F10 
                 0.936 
                 0.063 
                 0.084 
                 5.646 
                 0.432 
                 0.094 
                 0.126 
                 0.469 
                 1.350 
               
               
                   
                 P405.18.H5 
                 4.725 
                 0.219 
                 0.782 
                 1.100 
                 19.220 
                 0.027 
                 0.450 
                 43.684 
                 1.328 
               
               
                   
                 P405.19.A8 
                 0.291 
                 0.021 
                 &lt;0.0032 
                 0.116 
                 0.278 
                 0.059 
                 0.110 
                 0.034 
                 2.012 
               
               
                   
                 P405.19.B12 
                 0.889 
                 0.057 
                 0.093 
                 0.264 
                 1.103 
                 0.033 
                 0.233 
                 0.157 
                 0.807 
               
               
                   
                 P405.19.F11 
                 0.689 
                 0.018 
                 0.109 
                 2.892 
                 2.131 
                 0.037 
                 &gt;50 
                 5.279 
                 0.818 
               
               
                   
                 1140.6FS 
                 0.328 
                 0.029 
                 &lt;0.0032 
                 5.219 
                 6.915 
                 0.284 
                 &gt;50 
                 13.917 
                 21.450 
               
               
                   
                 1140.6G9 
                 0.748 
                 0.116 
                 0.114 
                 4.222 
                 0.096 
                 0.147 
                 &gt;50 
                 0.220 
                 6.710 
               
               
                 HISTORICAL 
                 P116.2 
                 5.406 
                 0 493 
                 0.422 
                 40.937 
                 5.647 
                 0.142 
                 &gt;50 
                 17.250 
                 16.580 
               
               
                   
                 P116.3.F6 
                 22.297 
                 0.235 
                 0.255 
                 0.495 
                 0.728 
                 0.158 
                 &gt;50 
                 15.152 
                 9.520 
               
               
                   
                 P116.3.G9 
                 1.054 
                 0.071 
                 0.018 
                 0.435 
                 3.157 
                 1.385 
                 0.540 
                 24.689 
                 5.750 
               
               
                   
                 P116.4.11 
                 2.594 
                 0.149 
                 0.353 
                 17.822 
                 6.646 
                 0.703 
                 0.329 
                 26.982 
                 13.790 
               
               
                   
                 1234.3A9 
                 2.623 
                 0.226 
                 0.032 
                 15.504 
                 3.944 
                 0.815 
                 0.062 
                 40.940 
                 24.800 
               
               
                   
                 1234.3O9 
                 0.563 
                 0.102 
                 0.057 
                 4.784 
                 0.539 
                 0.087 
                 0.178 
                 48.779 
                 15.780 
               
               
                   
                 658.8A6 
                 4.860 
                 0.355 
                 0.386 
                 &gt;50 
                 2.057 
                 0.379 
                 0.196 
                 31.416 
                 &gt;25 
               
               
                   
                 658.8D2 
                 2.832 
                 0.284 
                 0.201 
                 40.617 
                 1.651 
                 0.347 
                 0.197 
                 3.291 
                 &gt;25 
               
               
                   
                 658.8F8 
                 &lt;0.0032 
                 &lt;0 0032 
                 &lt;0.0032 
                 &lt;0.0032 
                 0.994 
                 0.142 
                 &gt;50 
                 0.018 
                 &gt;25 
               
               
                   
                 526.17-2C11 
                 &lt;0.0032 
                 0.032 
                 0.040 
                 0.028 
                 0.048 
                 0.123 
                 &gt;50 
                 0.049 
                 ND 
               
               
                   
                 526.17-2G1 
                 0.429 
                 0.141 
                 0.025 
                 &lt;0.0032 
                 5.002 
                 0.982 
                 &gt;50 
                 0.353 
                 ND 
               
               
                   
                 526.17-2G3 
                 2.825 
                 0.170 
                 0.120 
                 4.692 
                 0.687 
                 0.110 
                 &gt;50 
                 0.928 
                 ND 
               
               
                   
                 424.9F4 
                 0.065 
                 0.091 
                 0.016 
                 0.589 
                 2.534 
                 0.508 
                 &gt;50 
                 2.757 
                 ND 
               
               
                   
                 424.9H1 
                 17.101 
                 1.386 
                 1.114 
                 19.990 
                 4.508 
                 0.590 
                 &gt;50 
                 4.982 
                 ND 
               
               
                   
                 139.19A6 
                 &gt;50 
                 1.059 
                 1.091 
                 3.132 
                 &gt;50 
                 0.090 
                 &gt;50 
                 0.520 
                 ND 
               
               
                   
                 139.19.C10 
                 &gt;50 
                 0.118 
                 0.089 
                 5.745 
                 &gt;50 
                 0.019 
                 &gt;50 
                 0.093 
                 ND 
               
               
                   
                 139.19.F2 
                 &gt;50 
                 0.241 
                 0.226 
                 0.755 
                 11.125 
                 0.036 
                 &gt;50 
                 0.525 
                 ND 
               
               
                   
                 208.9.C6 
                 17.496 
                 0.375 
                 0.587 
                 25.217 
                 1.510 
                 0.587 
                 0.123 
                 &gt;50 
                 ND 
               
               
                   
                 208.9.F12 
                 5.263 
                 0.265 
                 0.314 
                 11.871 
                 4.460 
                 0.414 
                 0.234 
                 &gt;50 
                 ND 
               
               
                   
                 208.9.G10 
                 6.842 
                 0.151 
                 0.351 
                 2.896 
                 2.099 
                 0.832 
                 0.093 
                 &gt;50 
                 ND 
               
               
                   
                 1031.12.6C4 
                 0.701 
                 0.024 
                 0.083 
                 0.321 
                 9.717 
                 0.057 
                 &gt;50 
                 &gt;50 
                 17.730 
               
               
                   
                 1931.12.7D5 
                 0.140 
                 &lt;0.0032 
                 &lt;0.0032 
                 0.023 
                 19.210 
                 0.231 
                 &gt;50 
                 &gt;50 
                 13.770 
               
               
                   
                 1031.12.9D9 
                 &lt;0.0032 
                 &lt;0.0032 
                 &lt;0.0032 
                 &lt;0.3032 
                 0.071 
                 0.030 
                 &gt;50 
                 &gt;50 
                 18.220 
               
               
                   
                 1.7.1A7 
                 0.280 
                 0.116 
                 0.189 
                 &gt;50 
                 6.800 
                 0.176 
                 0.061 
                 0.743 
                 &gt;25 
               
               
                   
                 1.7.1D2 
                 10.695 
                 0.939 
                 0.998 
                 &gt;50 
                 &gt;50 
                 0.669 
                 0.062 
                 10.213 
                 &gt;25 
               
               
                   
                 1.7.1G10 
                 &gt;50 
                 0.199 
                 0.185 
                 0.81 
                 1.506 
                 0.745 
                 0.266 
                 0.307 
                 &gt;25 
               
               
                   
                 233.7.1B2 
                 &gt;50 
                 0.100 
                 0.212 
                 &gt;50 
                 &lt;0.0032 
                 0.158 
                 &lt;0.0032 
                 0.057 
                 15.000 
               
               
                   
                 233.7.1C3 
                 &gt;50 
                 0.973 
                 1.771 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 0.041 
                 0.132 
                 10.700 
               
               
                   
                 233.7.1C11 
                 &gt;50 
                 0.241 
                 0.226 
                 0.755 
                 11.125 
                 0.036 
                 &gt;50 
                 0.525 
                 13.370 
               
               
                   
                 458.5.12B1 
                 17.496 
                 0.375 
                 0.587 
                 25.217 
                 1.510 
                 0.587 
                 0.123 
                 &gt;50 
                 0.889 
               
               
                   
                 458.5.12E1 
                 5.263 
                 0.265 
                 0.314 
                 11.871 
                 4.460 
                 0.414 
                 0.234 
                 &gt;50 
                 2.110 
               
               
                   
                 458.5.12G9 
                 6.84 
                 0.151 
                 0.351 
                 2.396 
                 2.099 
                 0.832 
                 0.093 
                 &gt;50 
                 9.440 
               
               
                   
                 172.7C6 
                 0.701 
                 0.024 
                 0.089 
                 0.321 
                 9.717 
                 0.057 
                 &gt;50 
                 &gt;50 
                 ND 
               
               
                   
                 172.7F11 
                 0.140 
                 0.001 
                 0.001 
                 0.023 
                 19.210 
                 0.231 
                 &gt;50 
                 &gt;50 
                 ND 
               
               
                   
                 172.7G5 
                 0.001 
                 0.001 
                 0.001 
                 0.001 
                 0.071 
                 0.030 
                 &gt;50 
                 &gt;50 
                 ND 
               
               
                   
                 1161.9G11 
                 0.260 
                 0.116 
                 0.189 
                 &gt;50 
                 6.800 
                 0.176 
                 0.061 
                 0.743 
                 ND 
               
               
                   
                 1161.9C1 
                 10.695 
                 0.939 
                 0.998 
                 &gt;50 
                 &gt;50 
                 0.669 
                 0.062 
                 10.213 
                 ND 
               
               
                   
                 537.8.A11 
                 &gt;50 
                 0.199 
                 0.185 
                 0.871 
                 1.506 
                 0.745 
                 0.266 
                 0.307 
                 ND 
               
               
                   
                 537.8.E6 
                 &gt;50 
                 0.100 
                 3.212 
                 &gt;50 
                 0.001 
                 0.158 
                 0.001 
                 0.057 
                 ND 
               
               
                   
                 537.8.E10 
                 &gt;50 
                 0.973 
                 1.771 
                 &gt;50 
                 &gt;50 
                 &gt;50 
                 0.041 
                 0.132 
                 ND 
               
               
                   
               
               
                   
                 Virus ID 
                 b12 
                 2G12 
                 2F5 
                 PG9 
                 PG16 
                 VRC01 
                 45-46 54W 
                 PGT121 
                 10-1074 
               
               
                   
               
               
                 CONTEM- 
                 P035.6.E4 
                 11.865 
                 &lt;0.39 
                 3.480 
                 &gt;1 
                 &gt;1 
                 &lt;0.078 
                 0.032 
                 15.471 
                 0.059 
               
               
                 PORARY 
                 P035.6.H11 
                 10.758 
                 &lt;0.39 
                 2.115 
                 &gt;1 
                 &gt;1 
                 0.160 
                 0.018 
                 0.667 
                 0.143 
               
               
                   
                 P035.6.D10 
                 &gt;12.5 
                 &lt;0.39 
                 1.900 
                 &gt;1 
                 &gt;1 
                 0.390 
                 0.020 
                 0.174 
                 0.110 
               
               
                   
                 P151.37.C7 
                 1.832 
                 &gt;25 
                 3.350 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.076 
                 0.109 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P151.37.F1 
                 &gt;12.5 
                 &gt;25 
                 1.372 
                 &lt;0.116 
                 &lt;0.016 
                 0.250 
                 0.472 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P151.37.F10 
                 0.452 
                 &gt;25 
                 &lt;0.39 
                 &lt;0.016 
                 &lt;0.016 
                 1.160 
                 0.172 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P153.10.2.A9 
                 7.988 
                 0.825 
                 0.918 
                 &gt;1 
                 0.431 
                 &gt;5 
                 34.912 
                 42.623 
                 1.466 
               
               
                   
                 P153:10.2.D8 
                 &gt;12.5 
                 3.562 
                 24.193 
                 &gt;1 
                 0.853 
                 0.890 
                 0.325 
                 1.503 
                 0.620 
               
               
                   
                 P153.10.2.E10 
                 9.212 
                 &lt;0.39 
                 22.382 
                 &gt;1 
                 &lt;0.016 
                 &gt;5 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P186.12.1.D10 
                 9.333 
                 &gt;25 
                 2.492 
                 &gt;1 
                 &gt;1 
                 &gt;5 
                 22.865 
                 0.021 
                 0.212 
               
               
                   
                 P186.12.1.F4 
                 11.1375 
                 &gt;25 
                 15.277 
                 &gt;1 
                 &gt;1 
                 3.320 
                 15.929 
                 0.026 
                 0.094 
               
               
                   
                 P186.21.1.G2 
                 10.763 
                 21.986 
                 3.181 
                 &gt;1 
                 &gt;1 
                 &gt;5 
                 16.385 
                 0.018 
                 0.108 
               
               
                   
                 P195.31.A6 
                 0.498 
                 &gt;25 
                 3.884 
                 &gt;1 
                 &gt;1 
                 0.890 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P195.31.A10 
                 9.896 
                 &gt;25 
                 &lt;0.39 
                 &gt;1 
                 &gt;1 
                 4.800 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P195.31F11 
                 &gt;12.5 
                 &gt;25 
                 2.306 
                 &gt;1 
                 &gt;1 
                 1.930 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P019.1.D2 
                 0.432 
                 &lt;0.39 
                 &lt;0.39 
                 &lt;0.016 
                 &lt;0.016 
                 0.220 
                 0.051 
                 &gt;50 
                 0.191 
               
               
                   
                 P019.1.D8 
                 &gt;12.5 
                 9.371 
                 &lt;0.39 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.878 
                 0.043 
                 0.025 
                 0.528 
               
               
                   
                 P019.1.G7 
                 &gt;12.5 
                 6.764 
                 &lt;0.39 
                 0.050 
                 0.034 
                 &lt;0.078 
                 &lt;0.0032 
                 &gt;50 
                 0.318 
               
               
                   
                 P175.10.D7 
                 1.369 
                 &lt;0.39 
                 &gt;25 
                 &gt;1 
                 0.110 
                 1.100 
                 0.022 
                 0.068 
                 0.097 
               
               
                   
                 P175.10.D12 
                 &gt;12.5 
                 &lt;0.39 
                 &gt;25 
                 &gt;1 
                 0.150 
                 0.170 
                 0.032 
                 0.085 
                 0.241 
               
               
                   
                 P175.10.G10 
                 &gt;12.5 
                 2.183 
                 1.300 
                 0.720 
                 0.032 
                 1.660 
                 0.017 
                 0.069 
                 0.220 
               
               
                   
                 P013.18.A9 
                 &gt;12.5 
                 &gt;25 
                 2.763 
                 &gt;1 
                 &gt;1 
                 &gt;5 
                 0.083 
                 5144 
                 &gt;50 
               
               
                   
                 P154.44.C8 
                 &gt;12.5 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 0.600 
                 &gt;5 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P154.44.G8 
                 &gt;12.5 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 &gt;1 
                 0.790 
                 0.536 
                 1.996 
                 0.521 
               
               
                   
                 P183.50.2.H3 
                 5.923 
                 6.688 
                 1.527 
                 &gt;1 
                 &gt;1 
                 1.031 
                 0.195 
                 2.106 
                 0.185 
               
               
                   
                 P1833.2.B9 
                 1.230 
                 2.132 
                 1.073 
                 &gt;1 
                 &gt;1 
                 0.340 
                 0.056 
                 5.073 
                 0.266 
               
               
                   
                 P001.35.F5 
                 &gt;12.5 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 &gt;5 
                 0.946 
                 1.873 
                 0.046 
               
               
                   
                 P001.35 H4 
                 &gt;12.5 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 4.300 
                 0.051 
                 2.848 
                 0052 
               
               
                   
                 P002.39.CB 
                 &gt;12.5 
                 2.216 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.450 
                 1.129 
                 0.024 
               
               
                   
                 P002.39.F8 
                 &gt;12.5 
                 &lt;0.39 
                 17.778 
                 &lt;0.016 
                 &lt;0.016 
                 1.610 
                 0.625 
                 22.125 
                 0.049 
               
               
                   
                 P002.39.H10 
                 &gt;12.5 
                 &lt;0.39 
                 1.500 
                 &lt;0.016 
                 0.048 
                 3.030 
                 0.917 
                 1.985 
                 0.042 
               
               
                   
                 P034.6.D6 
                 11.122 
                 0.844 
                 0.873 
                 &lt;0.016 
                 0.122 
                 &lt;0.078 
                 0.859 
                 0.027 
                 &lt;0.0032 
               
               
                   
                 P034.6.G19 
                 8.598 
                 &lt;0.39 
                 1.257 
                 &gt;1 
                 &gt;1 
                 &gt;5 
                 &gt;50 
                 &gt;50 
                 0.028 
               
               
                   
                 P034.6 H5 
                 &gt;12.5 
                 &gt;25 
                 1.163 
                 0.017 
                 0.021 
                 0.310 
                 0.127 
                 0.037 
                 0.018 
               
               
                   
                 P101.20.1.F1 
                 &gt;12.5 
                 &gt;25 
                 2.721 
                 &gt;1 
                 &gt;1 
                 2.420 
                 0.676 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 P101.20.1 HB 
                 &gt;12.5 
                 &gt;25 
                 1.719 
                 &gt;1 
                 &gt;1 
                 1.470 
                 0.861 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 P127.46.A6 
                 6.780 
                 2.345 
                 1.671 
                 0.079 
                 &lt;0.016 
                 &lt;0.078 
                 0.143 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P127.46.D1 
                 &gt;12.5 
                 &lt;0.39 
                 0.921 
                 0.101 
                 0.150 
                 0.230 
                 0.203 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P127.45.D2 
                 &lt;0.195 
                 &gt;25 
                 1.126 
                 0.378 
                 &lt;0.016 
                 &lt;0.978 
                 0.023 
                 &gt;50 
                 &gt;50 
               
               
                   
                 P174.28.E11 
                 8.149 
                 0.074 
                 1.574 
                 0.023 
                 &lt;0.016 
                 3.910 
                 1.863 
                 0.085 
                 0.050 
               
               
                   
                 P177.25.1.G9 
                 &gt;12.5 
                 &gt;25 
                 5.851 
                 &gt;1 
                 &gt;1 
                 &lt;0.078 
                 0.325 
                 0.128 
                 0.034 
               
               
                   
                 P177.25.2.B4 
                 &gt;12.5 
                 &gt;25 
                 1.241 
                 &gt;1 
                 &gt;1 
                 0.450 
                 0.530 
                 0.029 
                 0.023 
               
               
                   
                 P177.25.2.D1 
                 7.770 
                 &gt;25 
                 1.232 
                 &gt;1 
                 &gt;1 
                 3.630 
                 0.066 
                 0.017 
                 0.018 
               
               
                   
                 P180.14.A6 
                 &gt;12.5 
                 17.668 
                 &gt;25 
                 0.038 
                 &lt;0.016 
                 1.000 
                 2 094 
                 0.139 
                 0.028 
               
               
                   
                 P130.14.G6 
                 &gt;12.5 
                 2.988 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 2.380 
                 0.306 
                 0.174 
                 0.020 
               
               
                   
                 P180.14.G7 
                 &gt;12.5 
                 2.400 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 1.240 
                 1.012 
                 0.028 
                 0.038 
               
               
                   
                 P197.251.D2 
                 &gt;12.5 
                 &gt;12.5 
                 1.224 
                 &lt;0.016 
                 &lt;0.016 
                 0.090 
                 0.025 
                 14.287 
                 0.095 
               
               
                   
                 P197.25.1.D7 
                 &gt;12.5 
                 1.232 
                 1.291 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.052 
                 20.337 
                 0.057 
               
               
                   
                 P197.25.1.H1 
                 &gt;12.5 
                 1.079 
                 2.088 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.072 
                 23.417 
                 0.092 
               
               
                   
                 P405.18.D3 
                 0.471 
                 &lt;0.39 
                 1.137 
                 &gt;1 
                 &gt;1 
                 &gt;5 
                 0.254 
                 0.071 
                 0.018 
               
               
                   
                 P405.18.F10 
                 0.716 
                 &lt;0.39 
                 0.804 
                 ND 
                 ND 
                 &gt;5 
                 &gt;50 
                 5.680 
                 &gt;50 
               
               
                   
                 P405.18.H5 
                 0.497 
                 0.386 
                 0.815 
                 0.141 
                 &lt;0.016 
                 &gt;5 
                 &gt;50 
                 4.760 
                 0.187 
               
               
                   
                 P405.19.A8 
                 1.116 
                 0.814 
                 0.878 
                 &gt;1 
                 0.044 
                 0.470 
                 0.031 
                 0.917 
                 &lt;0.0032 
               
               
                   
                 P405.19.B12 
                 0.656 
                 0.986 
                 1.029 
                 0.190 
                 &lt;0.016 
                 &gt;5  
                 &lt;0.0032 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 P405.19.F11 
                 0.413 
                 &lt;0.39 
                 3.630 
                 0.190 
                 0.034 
                 &gt;5 
                 ND 
                 ND 
                 ND 
               
               
                   
                 1140.6FS 
                 &gt;25 
                 &gt;25 
                 3.790 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 &lt;0.0032 
                 0.017 
                 0.019 
               
               
                   
                 1140.6G9 
                 3.270 
                 &gt;25 
                 1.520 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.019 
                 0.112 
                 0.024 
               
               
                 HISTORICAL 
                 P116.2 
                 17.370 
                 &gt;25 
                 4.170 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 &lt;0.0032 
                 0.017 
                 &lt;0.0032 
               
               
                   
                 P116.3.F6 
                 13.540 
                 &gt;25 
                 3.030 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.020 
                 0.019 
                 &lt;0.0032 
               
               
                   
                 P116.3.G9 
                 0.650 
                 &gt;25 
                 3.460 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 &lt;0.0032 
                 0.033 
                 &lt;0.0032 
               
               
                   
                 P116.4.11 
                 3.120 
                 &gt;25 
                 7.860 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 &lt;0.0032 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 1234.3A9 
                 &gt;25 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 &lt;0.0032 
                 0.022 
                 &lt;0.0032 
               
               
                   
                 1234.3O9 
                 &gt;25 
                 &gt;25 
                 8.550 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.028 
                 0.022 
                 &lt;0.0032 
               
               
                   
                 658.8A6 
                 &gt;25 
                 &gt;25 
                 &gt;25 
                 0.142 
                 0.032 
                 &lt;0.078 
                 &lt;0.0032 
                 0.151 
                 0.049 
               
               
                   
                 658.8D2 
                 2.000 
                 &gt;25 
                 17.210 
                 0.135 
                 &lt;0.016 
                 &lt;0.078 
                 &lt;0.0032 
                 0.051 
                 0.029 
               
               
                   
                 658.8F8 
                 &gt;25 
                 &gt;25 
                 &gt;25 
                 0.039 
                 &lt;0.016 
                 &lt;0.078 
                 &lt;0.0032 
                 0.026 
                 0.017 
               
               
                   
                 526.17-2C11 
                 ND 
                 ND 
                 ND 
                 &lt;0.016 
                 &lt;0.016 
                 0.199 
                 0.046 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 526.17-2G1 
                 ND 
                 ND 
                 ND 
                 0.059 
                 &lt;0.016 
                 0.840 
                 0.074 
                 &lt;0.0032 
                 0.018 
               
               
                   
                 526.17-2G3 
                 ND 
                 ND 
                 ND 
                 &lt;0.016 
                 &lt;0.016 
                 0.354 
                 &lt;0.0032 
                 0.087 
                 &lt;0.0032 
               
               
                   
                 424.9F4 
                 ND 
                 ND 
                 ND 
                 &lt;0.016 
                 &lt;0.016 
                 0.310 
                 0.028 
                 0.032 
                 0.019 
               
               
                   
                 424.9H1 
                 ND 
                 ND 
                 ND 
                 0.039 
                 &lt;0.016 
                 &gt;5 
                 0.369 
                 3.731 
                 1.087 
               
               
                   
                 139.19A6 
                 ND 
                 ND 
                 ND 
                 &lt;0.016 
                 &lt;0.016 
                 0.557 
                 0.127 
                 0.066 
                 0.017 
               
               
                   
                 139.19.C10 
                 ND 
                 ND 
                 ND 
                 &lt;0.016 
                 &lt;0.016 
                 1.549 
                 0.888 
                 0.118 
                 0.039 
               
               
                   
                 139.19.F2 
                 ND 
                 ND 
                 ND 
                 &lt;0.016 
                 &lt;0.016 
                 &gt;5 
                 0.669 
                 0.106 
                 0.039 
               
               
                   
                 208.9.C6 
                 ND 
                 ND 
                 ND 
                 0.063 
                 0.248 
                 0.303 
                 0.100 
                 0.125 
                 0.137 
               
               
                   
                 208.9.F12 
                 ND 
                 ND 
                 ND 
                 &gt;1 
                 &gt;1 
                 0.714 
                 0.128 
                 0.086 
                 0.151 
               
               
                   
                 208.9.G10 
                 ND 
                 ND 
                 ND 
                 0.882 
                 &gt;1 
                 0.289 
                 0.083 
                 &lt;0.0032 
                 0.023 
               
               
                   
                 1031.12.6C4 
                 1.180 
                 &gt;25 
                 6.280 
                 &gt;1 
                 &gt;1 
                 0.160 
                 0.040 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 1931.12.7D5 
                 0.960 
                 &gt;25 
                 8.500 
                 &gt;1 
                 &gt;1 
                 0.120 
                 0.072 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 1031.12.9D9 
                 3.540 
                 &gt;25 
                 10.770 
                 &gt;1 
                 &gt;1 
                 0.200 
                 0.102 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 1.7.1A7 
                 &gt;25 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 0.640 
                 ND 
                 ND 
                 ND 
               
               
                   
                 1.7.1D2 
                 11.920 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 0.700 
                 0.300 
                 0.045 
                 0.048 
               
               
                   
                 1.7.1G10 
                 &gt;25 
                 &gt;25 
                 &gt;25 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.282 
                 0.017 
                 &lt;0.0032 
               
               
                   
                 233.7.1B2 
                 &gt;25 
                 0.140 
                 5.030 
                 0.019 
                 &lt;0.016 
                 0.160 
                 0.019 
                 0.030 
                 &lt;0.0032 
               
               
                   
                 233.7.1C3 
                 8.030 
                 0.330 
                 5.900 
                 0.528 
                 &gt;1 
                 &lt;0.078 
                 0.027 
                 &lt;0.0032 
                 &lt;0.0032 
               
               
                   
                 233.7.1C11 
                 18.480 
                 0.870 
                 5.460 
                 &gt;1 
                 0.370 
                 0.090 
                 0.024 
                 0.023 
                 0.038 
               
               
                   
                 458.5.12B1 
                 9.990 
                 &gt;25 
                 3.829 
                 0.028 
                 0.311 
                 1.370 
                 0.035 
                 0.018 
                 0.018 
               
               
                   
                 458.5.12E1 
                 1.850 
                 22.620 
                 1.660 
                 &lt;0.016 
                 &lt;0.016 
                 &lt;0.078 
                 0.072 
                 0.201 
                 0.020 
               
               
                   
                 458.5.12G9 
                 4.910 
                 &gt;25 
                 2.350 
                 &gt;1 
                 0.039 
                 0.100 
                 0.223 
                 0.022 
                 0.023 
               
               
                   
                 172.7C6 
                 ND 
                 ND 
                 ND 
                 0.020 
                 &lt;0.016 
                 0.431 
                 0.019 
                 0.025 
                 0.027 
               
               
                   
                 172.7F11 
                 ND 
                 ND 
                 ND 
                 0.017 
                 &lt;0.016 
                 0.424 
                 0.021 
                 0.020 
                 0.025 
               
               
                   
                 172.7G5 
                 ND 
                 ND 
                 ND 
                 0.048 
                 &lt;0.016 
                 &lt;0.378 
                 &lt;0.0032 
                 0.035 
                 &lt;0.0032 
               
               
                   
                 1161.9G11 
                 ND 
                 ND 
                 ND 
                 &gt;1 
                 &gt;1 
                 0.378 
                 0.021 
                 &lt;0.0032 
                 0.020 
               
               
                   
                 1161.9C1 
                 ND 
                 ND 
                 ND 
                 &gt;1 
                 &gt;1 
                 &gt;5 
                 0.618 
                 0.065 
                 0.051 
               
               
                   
                 537.8.A11 
                 ND 
                 ND 
                 ND 
                 &gt;1 
                 &gt;1 
                 3.459 
                 0.270 
                 0.031 
                 0.021 
               
               
                   
                 537.8.E6 
                 ND 
                 ND 
                 ND 
                 &gt;1 
                 &gt;1 
                 0.331 
                 0.065 
                 0.086 
                 0.027 
               
               
                   
                 537.8.E10 
                 ND 
                 ND 
                 ND 
                 &gt;1 
                 &gt;1 
                 2.071 
                 0.174 
                 0.025 
                 0.017 
               
               
                   
               
               
                 Numbers indicate antibody lgG concentrations in μg/ml to reach the IC 50  in the PBMC-based neutralization assay. 
               
               
                 IC 50  values indicate an increasing neutralization sensitivity. 
               
               
                 &gt;indicates that the IC 50  for a given virus was not reached at the concetration tested. 
               
               
                 ND, not determined. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Data collection and refinement statistics (molecular replacement) 
               
            
           
           
               
               
               
               
               
            
               
                   
                 PGT121 Fab 
                   
                   
                 PGT121 Fab  
               
               
                   
                 “unliganded” 
                 10-1074 Fab 
                 GL Fab 
                 “llganded” 
               
               
                   
               
               
                 Data collection 
                   
                   
                   
                   
               
               
                 Space group 
                 P2 1 2 1 2 1   
                 P2 1   
                 P2 1   
                 P2 1 2 1 2 1   
               
               
                 Cell dimensions 
                   
                   
                   
                   
               
               
                 a, b, c (Å) 
                 56.75, 74.67, 114.917 
                 61.38, 40.26, 84.46 
                 54.93, 344.74, 55.23 
                 67,79, 67.79, 94.11 
               
               
                 α, β, γ (°) 
                 90.00, 90.00, 90.00 
                 90.00, 95.39, 90.00 
                 90.00, 91.95, 90.00 
                 90.00, 90.00, 90.00 
               
               
                 Resolution (Å) 
                 2.78-35.5 (2.78-2.93) 
                 1.80- 36.31 (1,80-1.91) 
                 2.42-38.60 (2.42-2.55) 
                 2.33-38.66 (2.33-2.47) 
               
               
                 R merge   
                 0.099 (0.293) 
                 0.075 (0.558) 
                 0.072 (0.482) 
                 0.161 (0.603) 
               
               
                 I/σ i   
                 8.8 (3.1) 
                 8.7 (1.8) 
                 11.0 (1.9)  
                 8.7 (2.9) 
               
               
                 Completeness (%) 
                 96.7 (84.8) 
                 93.49 (98.0)  
                 95.5 (80.1) 
                 92.2 (98.9) 
               
               
                 Redundancy 
                 3.2 (2.7) 
                 2.7 (2.8) 
                 3.1 (2.6) 
                 5.3 (5.8) 
               
               
                 Refinement 
                   
                   
                   
                   
               
               
                 Resolution (Å) 
                 3.0 
                 1.9 
                 2.42 
                 2.4 
               
               
                 No. reflections 
                 10,076 
                 31,363 
                 74,237 
                 16,831 
               
               
                 R work /R free   
                 0.216/0.264 
                 0.187/0.223 
                 0.194/0.237 
                 0.201/0.249 
               
               
                 No. atoms 
                   
                   
                   
                   
               
               
                 Protein 
                 3,276 
                 3,346 
                 12,881 
                 3,127 
               
               
                 Ligand/ion 
                 0 
                 0 
                 0 
                 129 
               
               
                 Water 
                 0  
                 300 
                 527 
                 203 
               
               
                 B-factors 
                   
                   
                   
                   
               
               
                 Protein 
                 32.78 
                 29.17 
                 44.67 
                 31.48 
               
               
                 Ligand/ion 
                 — 
                 — 
                 — 
                 45.1 
               
               
                 Water 
                 — 
                 37.37 
                 40.27 
                 36.78 
               
               
                 R.m.s. deviations 
                   
                   
                   
                   
               
               
                 Bond lengths (Å) 
                 0.005 
                 0.007 
                 0.005 
                 0.006 
               
               
                 Bond angles (°) 
                 0.971 
                 1.234 
                 0.951 
                 0.949 
               
               
                   
               
               
                 *Data for each structure were acquired from a single crystal. 
               
               
                 *Values in parentheses are for the highest-resolution shell. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 RMSD values for Cα alignments of Fabs 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Fab1/Fab2 
                 RMSD VH  (Å) 
                 # residues 
                 RMSD VL  (Å) 
                 # residues 
                 RMSD VH+VL  (Å) 
                 # residues 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 PGT121/PGT128 
                 1.159 
                 116/130 
                 1.63 
                 95/100 
                 1.462 
                 207/235 
               
               
                 PGT121/PGT145 
                 2.93 
                 124/130 
                 1.91 
                 94/105 
                 1.75 
                 206/235 
               
               
                 PGT121/10-1074 
                 0.74 
                 128/130 
                 1.2 
                 102/105  
                 1.26 
                 226/235 
               
               
                 PGT121/GL 
                 1.33 
                 129/130 
                 1.37 
                 94/105 
                 1.6 
                 225/235 
               
               
                 10-1074/GL 
                 1.38 
                 130/130 
                 1.35 
                 92/105 
                 1.39 
                 220/235 
               
               
                 PGT121/PGT12 liganded   
                 0.79 
                 125/128 
                 0.5 
                 100/100 
                 0.78 
                 225/228 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Contacts between PGT121 Fab and bound glycan 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 Glycan atom 
                 Protein atom 
                 Water 
                 Distance (Å) 
                 Glycan atom 
                 Protein atom 
                 Water 
                 Distance (Å) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 GlcNAc 6 -O3 
                 Asn 58 -Nδ2 
                   
                 2.91 
                 Sia 10 -O8 
                 Asp 31 -O 
                   
                 2.72 
               
               
                 GlcNAc 6 -O7 
                 Asn 58 -Oδ1 
                   
                 2.94 
                 Sia 10 -O10 
                 His 97 -N 
                   
                 3.18 
               
               
                 GlcNAc 6 -O6 
                   
                 H 2 O 471   
                 3.15 
                 Sia 10 -O10 
                 His 97 -O 
                   
                 3.19 
               
               
                 GlcNAc 6 -O4 
                   
                 H 2 O 477   
                 3.05 
                 Sia 10 -O9 
                   
                 H 2 O 480   
                 3.19 
               
               
                 GlcNAc 6 -O3 
                   
                 H 2 O 481   
                 2.94 
                 Sia 10 -O8 
                 Ser 32 -Oγ 
                   
                 3.70* 
               
               
                 Man 1 -O4 
                   
                 H 2 O 410   
                 3.02 
                 Man 3 -O3 
                 Asn 56 -Oδ1 
                   
                 2.58 
               
               
                 Man 1 -O4 
                   
                 H 2 O 420   
                 2.66 
                 Man 3 -O6 
                   
                 H 2 O 477   
                 3.35 
               
               
                 Man 1 -O3 
                   
                 H 2 O 410   
                 3.35 
                 GlcNAc 4 -O5 
                 Thr 57 -O 
                   
                 3.33 
               
               
                 Man 1 -O2 
                   
                 H 2 O 477   
                 3.14 
                 GlcNAc 4 -N2 
                   
                 H 2 O 479   
                 3.2 
               
               
                 Man 1 -O5 
                   
                 H 2 O 477   
                 2.62 
                 Fuc 9 -O2 
                   
                 H 2 O 471   
                 2.57 
               
               
                 Man 2 -O6 
                 Thr 100 -Oγ1 
                   
                 3.34 
                   
                 Asp 31 -O 
                 H 2 O 435   
                 3.09 
               
               
                 Man 2 -O2 
                   
                 H 2 O 410   
                 3.41 
                   
                 Asp 31 -Oδ1 
                 H 2 O 480   
                 3.32 
               
               
                 Man 2 -O5 
                   
                 H 2 O 446   
                 2.95 
                   
                 Tyr 50 -OH 
                 H 2 O 481   
                 2.8 
               
               
                 Man 2 -O6 
                   
                 H 2 O 446   
                 3.26 
                   
                 His 52 -Nε2 
                 H 2 O 435   
                 3.16 
               
               
                 GlcNAc 7 -N2 
                 Tyr 33 -OH 
                   
                 2.72 
                   
                 Ser 54 -Oγ 
                 H 2 O 419   
                 3.2 
               
               
                 GlcNAc 7 -O5 
                   
                 H 2 O 410   
                 3.38 
                   
                 Ser 54 -O 
                 H 2 O 420   
                 3.16 
               
               
                 GlcNAc 7 -O7 
                   
                 H 2 O 411   
                 3.00 
                   
                 Gly 55 -O 
                 H 2 O 479   
                 2.85 
               
               
                 GlcNAc 7 -O3 
                 His 97 -Nδ2 
                   
                 3.60* 
                   
                 Asp 56 -Oδ1 
                 H 2 O 481   
                 3.49 
               
               
                 GlcNAc 7 -O7 
                 His 97 -Nε2 
                   
                 3.70* 
                   
                 Asp 56 -Oδ2 
                 H 2 O 446   
                 3.07 
               
               
                 Gal 8 -O3 
                 Lys 53 -Nζ 
                   
                 2.97 
                   
                 Asn 58 -Nδ2 
                 H 2 O 481   
                 3.15 
               
               
                 Gal 8 -O4 
                   
                 H 2 O 480   
                 3.47 
                   
                 Arg 99 -Nε 
                 H 2 O 411   
                 2.58 
               
               
                 Gal 8 -O4 
                   
                 H 2 O 435   
                 2.76 
                   
                 Thr 100 -Oγ1 
                 H 2 O 411   
                 2.94 
               
               
                 Gal 8 -O5 
                   
                 H 2 O 435   
                 3.17 
               
               
                   
               
               
                 Hydrogen bond criteria: bond distance &lt; 3.5 A, O—H—O/N—-H—O angle &gt; 90°  
               
               
                 *Contacts are close to hydrogen bond distance cutoff and are included as possible interactions 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 In vitro neutralization activity of PGT121GM and 10-1074GM 
               
            
           
           
               
               
               
               
               
               
            
               
                 Virus ID 
                 Clade 
                 PGT121 
                 PGT121GM 
                 10-1074 
                 10-1074GM 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Q842.d12 
                 A 
                 0.074 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 3365.v2.c2 
                 A 
                 7.353 
                 &gt;50 
                 0.450 
                 0.467 
               
               
                 0260.v5.c36 
                 A 
                 0.152 
                 &gt;50 
                 0.160 
                 0.618 
               
               
                 YU.2 
                 B 
                 0.356 
                 1.355 
                 0.398 
                 0.262 
               
               
                 TRO.11 
                 B 
                 0.051 
                 0.258 
                 0.057 
                 0.049 
               
               
                 TRJO4551.58 
                 B 
                 35.291 
                 &gt;50 
                 0.634 
                 0.721 
               
               
                 QH0692.42 
                 B 
                 8.545 
                 &gt;50 
                 0.929 
                 0.376 
               
               
                 PVO.4 
                 B 
                 0.945 
                 47.564 
                 0.360 
                 0.138 
               
               
                 RHPA4259.7 
                 B 
                 0.054 
                 20.801 
                 0.118 
                 0.087 
               
               
                 WITO4160.33 
                 B 
                 6.007 
                 &gt;50 
                 2.112 
                 0.406 
               
               
                 1054_07_TC4_1499 
                 B (T/F) 
                 0.696 
                 &gt;50 
                 0.563 
                 0.193 
               
               
                 6244_13_65_4576 
                 B (T/F) 
                 1.878 
                 46.680 
                 0.922 
                 0.394 
               
               
                 62357_14_D3_4589 
                 B (T/F) 
                 45.559 
                 &gt;50 
                 &gt;50 
                 40.782 
               
               
                 CNE19 
                 BC 
                 0.189 
                 48.092 
                 50 
                 0.379 
               
               
                 CNE17 
                 BC 
                 &gt;50 
                 &gt;50 
                 13.297 
                 4.816 
               
               
                 CNE58 
                 BC 
                 &gt;50 
                 &gt;50 
                 0.968 
                 1.158 
               
               
                 CNE30 
                 BC 
                 0.559 
                 8.401 
                 1.200 
                 1.045 
               
               
                 CNE52 
                 BC 
                 32.935 
                 &gt;50 
                 13.147 
                 6.664 
               
               
                 ZM233M.PB6 
                 C 
                 8.977 
                 &gt;50 
                 0.349 
                 0.232 
               
               
                 ZM53M.PB12 
                 C 
                 0.002 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 CAP45.2.00.G3 
                 C 
                 6.544 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 HIV-16055-2.3 
                 C 
                 4.290 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 HIV-16845-2.22 
                 C 
                 &gt;50 
                 &gt;50 
                 5.835 
                 2.678 
               
               
                 ZM214M.PL15 
                 C 
                 3.150 
                 &gt;50 
                 2.367 
                 0.200 
               
               
                 ZM135M.PL10a 
                 C 
                 5.885 
                 &gt;50 
                 0.367 
                 0.184 
               
               
                 Ce1086_B2 
                 C (T/F) 
                 0.006 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 Ce1172_H1 
                 C (T/F) 
                 0.088 
                 0.180 
                 0.166 
                 0.054 
               
               
                 1394C9G1 (Rev-) 
                 C (T/F) 
                 3.372 
                 2.120 
                 0.191 
                 0.075 
               
               
                 3817.v2.c59 
                 CD 
                 &gt;50 
                 &gt;50 
                 14.880 
                 3.423 
               
               
                 6952.v1.c20 
                 CD 
                 0.605 
                 &gt;50 
                 0.138 
                 0.134 
               
               
                 BJOX009000.02.4 
                 CRF01_AE 
                 37.289 
                 &gt;50 
                 &gt;50 
                 &gt;50 
               
               
                 211-9 
                 CRF02_AG 
                 8.840 
                 &gt;50 
                 0.425 
                 0.976 
               
               
                 928-28 
                 CRF02_AG 
                 &gt;50 
                 &gt;50 
                 4.696 
                 3.121 
               
               
                 T251-18 
                 CRF02_AG 
                 &gt;50 
                 &gt;50 
                 7.395 
                 3.459 
               
               
                 T278-50 
                 CRF02_AG 
                 &gt;50 
                 &gt;50 
                 18.276 
                 12.017 
               
               
                 263-8 
                 CRF02_AG 
                 24.576 
                 &gt;50 
                 6.527 
                 7.779 
               
               
                 235-47 
                 CRF02_AG 
                 1.676 
                 &gt;50 
                 0.163 
                 0.069 
               
               
                 A07412m1.vrv12 
                 D 
                 0.406 
                 16.947 
                 0.048 
                 0.044 
               
               
                 X1193c1 
                 G 
                 0.202 
                 11.859 
                 0.475 
                 0.195 
               
               
                 X1254_c3 
                 G 
                 0.199 
                 0.222 
                 0.297 
                 0.112 
               
               
                   
               
               
                 Numbers indicate antibody 1 gG concentrations in μg/mi to reach the ICso in the TZM-bl neutralization assay. IC 50  values indicate an increasing neutralization sensitivity. 
               
               
                 &gt;indicates that the IC 50  for a given virus was not reached at the concentration tested. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 13 
               
             
            
               
                   
               
               
                 SHIV AD6EO   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Abs cone 
                 Titer 
               
               
                   
                   
                   
                   
                 (μg/ml) 
                 (TZM-bt) 
               
               
                 Animal ID 
                 Abs 
                 Dosage 
                 PROTECTED 
                 at Day 0 
                 at Day 0 
               
               
                   
               
               
                 RHDEGF 
                 VSC01 
                   50 mg/Kg 
                 Yes 
                 586.9  
                  1:162 
               
               
                 RHDEH3 
                   
                   
                 No 
                 711.0  
                  1:176 
               
               
                 RHDE1L 
                   
                   20 mg/Kg 
                 No 
                 206.5  
                  1:65 
               
               
                 RHJ8N 
                   
                   
                 No 
                 188.1  
                  1:68 
               
               
                 RHKNX 
                 PGT121 
                   20 mg/Kg 
                 Yes 
                 267.9  
                   1:2495 
               
               
                 RHMK4 
                   
                   
                 Yes 
                 253.6  
                   1:2773 
               
               
                 RHDE9J 
                   
                   5 mg/Kg 
                 Yes 
                 55.7  
                  1:563 
               
               
                 RHPNR 
                   
                   
                 No 
                 47.2  
                  1:618 
               
               
                 RHDCGI 
                   
                   1 mg/Kg 
                 Yes 
                 24.0  
                  1:116 
               
               
                 RHKNE 
                   
                   
                 Yes 
                 19.7  
                  1:55 
               
               
                 RHK44 
                   
                  0.2 mg/Kg 
                 No 
                 1.8 
                 &lt;1:20 
               
               
                 RHK49 
                   
                   
                 No 
                 1.8 
                  1:17 
               
               
                 RHDEEM 
                 10-1074 
                   20 mg/Kg 
                 Yes 
                 289.8  
                   1:2004 
               
               
                 RHKIL 
                   
                   
                 Yes 
                 257.7  
                   1:2075 
               
               
                 RHME1 
                   
                   5 mg/Kg 
                 Yes 
                 112.9  
                  1:633 
               
               
                 RHPNV 
                   
                   
                 Yes 
                 117.5  
                  1:384 
               
               
                 RHPID 
                   
                   1 mg/Kg 
                 No 
                 19.9  
                  1:56 
               
               
                 RHDCHX 
                   
                   
                 No 
                 24.8  
                  1:53 
               
               
                 RHPZE 
                 3BNC117 
                   5 mg/Kg 
                 Yes 
                 105.4  
                  1:272 
               
               
                 RHPM5 
                   
                   
                 Yes 
                 76.1  
                  1:372 
               
               
                 RHKMH 
                   
                   1 mg/Kg 
                 No 
                 39.6  
                  1:55 
               
               
                 RHMJ5 
                   
                   
                 No 
                 15.1  
                  1:75 
               
               
                 RHPLD 
                 45-46m2 
                   20 mg/Kg 
                 No 
                 15.0  
                  1:27 
               
               
                 RHMA9 
                   
                   
                 No 
                 17.6  
                 &lt;1:20 
               
               
                 RHMC6 
                   
                   5 mg/Kg 
                 No 
                 2.3 
                 &lt;1:20 
               
               
                 RHDE0CA 
                   
                   
                 No 
                 2.2 
                 &lt;1:20 
               
               
                 RHML1 
                 DEN3 
                   20 mg/Kg 
                 No 
                 ND 
                 &lt;1:20 
               
               
                 RHMAA 
                   
                   
                 No 
                 ND 
                 &lt;1:20 
               
               
                 RHDEJ3 
                 VRC01 
                   30 mg/Kg 
                 Yes 
                 395.8  
                  1:52 
               
               
                 RHKZ1 
                   
                   
                 No 
                 306.0  
                  1:70 
               
               
                 RHKZA 
                 PGT121 
                   20 mg/Kg 
                 Yes 
                 215.1  
                   1:13120 
               
               
                 RHDECT 
                   
                   
                 Yes 
                 200.7  
                   1:13805 
               
               
                 RHKTL 
                   
                   
                 Yes 
                 282.7  
                   1:12669 
               
               
                 RHPZ9 
                   
                   
                 Yes 
                 133.1  
                   1:12055 
               
               
                 RHK2Z 
                   
                   1 mg/Kg 
                 Yes 
                 15.1  
                  1:422 
               
               
                 RHMT8 
                   
                   
                 No 
                 29.3  
                  1:539 
               
               
                 RHDEE B 
                   
                  0.2 mg/Kg 
                 Yes 
                 3.1 
                  1:159 
               
               
                 RHDEP2 
                   
                   
                 Yes 
                 1.6 
                  1:101 
               
               
                 RHMFD 
                   
                 0.05 mg/Kg 
                 No 
                 1.0 
                 &lt;1:20 
               
               
                 RHKIA 
                   
                   
                 Na 
                 1.3 
                 &lt;1:20 
               
               
                 RHKIM 
                 10-1074 
                   20 mg/Kg 
                 Yes 
                 290.3  
                   1:1972 
               
               
                 RHKWM 
                   
                   
                 Yes 
                 173.3  
                   1:2282 
               
               
                 RHMJW 
                   
                   5 mg/Kg 
                 Yes 
                 96.6  
                  1:420 
               
               
                 RHMJT 
                   
                   
                 Yes 
                 95.3  
                  1:376 
               
               
                 RHDENI 
                   
                   1 mg/Kg 
                 Yes 
                 28.4  
                  1:106 
               
               
                 RHJHZ 
                   
                   
                 No 
                 18.6  
                  1:136 
               
               
                 RHHE8 
                   
                  0.2 mg/Kg 
                 No 
                 19.4  
                  1:39 
               
               
                 RHKCZ 
                   
                   
                 No 
                 19.7  
                  1:35 
               
               
                 RHMFBA 
                 3BNC117 
                   20 mg/Kg 
                 Yes 
                 294.9  
                  1:143 
               
               
                 RHMER 
                   
                   
                 Yes 
                 272.7  
                  1:142 
               
               
                 RHKIV 
                   
                   5 mg/Kg 
                 Yes 
                 114.8  
                  1:80 
               
               
                 RHKPI 
                   
                   
                 No 
                 133.1  
                  1:90 
               
               
                 RHDE9D 
                   
                   1 mg/Kg 
                 No 
                 23.3  
                  1:20 
               
               
                 RHDEW7 
                   
                   
                 Yes 
                 29.6  
                  1:18 
               
               
                 RHMEV 
                   
                  0.2 mg/Kg 
                 No 
                 3.9 
                 &lt;1:20 
               
               
                 RHMF9 
                   
                   
                 No 
                 5.7 
                 &lt;1:20 
               
               
                 RHKZMA 
                 45-46m2 
                   5 mg/Kg 
                 No 
                 2.1 
                 ND 
               
               
                 RHKNP 
                   
                   
                 No 
                 4.0 
                 ND 
               
               
                 RHJII 
                 hu-IgG 
                  100 mg/Kg 
                 No 
                 ND 
                 ND 
               
               
                 RHJKI 
                   
                   
                 No 
                 ND 
                 ND 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
             
               
                 TABLE 14 
               
             
            
               
                   
               
               
                   
                 IC 50  in TZM-b1 cells 2   
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                   
                   
                   
                 HIVIG 
                 Tier 
               
               
                 Sample ID 
                 S321 
                 C500 
                 B520 
                 G435 
                 T520b 
                 M263 
                 M600c 
                 (μg/ml) 
                 Phenotype 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 R5 SHIV DH12-V3AD8   
                 321 
                 289 
                 77 
                 172 
                 168 
                 429 
                 134 
                 132 
                 2 
               
               
                 R5 SHIV AD8-EO   
                 48 
                 36 
                 39 
                 31 
                 41 
                 44 
                 48 
                 1768 
                 2 
               
               
                 X4 SHIV DH12-CL7   
                 110 
                 94 
                 50 
                 65 
                 109 
                 115 
                 65 
                 530 
                 2 
               
               
                 HIV-1 CAAN5342.A2   
                 84 
                 &lt;20 
                 27 
                 &lt;10 
                 &lt;20 
                 77 
                 185 
                 638 
                 1 
               
               
                 HIV-1 MN.3   
                 13944 
                 9152 
                 821 
                 8431 
                 3968 
                 43722 
                 1709 
                 1.81 
                 1 
               
               
                   
               
               
                   2 Values are the serum dilution at which relative luminescence units (RLUs) were reduced 50% compared to virus control wells (no test sample). 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 15 
               
             
            
               
                   
               
               
                 SHIVAD8EO 
               
            
           
           
               
               
               
               
            
               
                 Endpoint 
                   
                   
                   
               
               
                 neutral- 
                   
                   
                   
               
               
                 ization 
                   
                   
                   
               
               
                 titer in 
                 Animal number 
                 Accumulated value 
                 Protected 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 plasma 
                 Protected 
                 Infected 
                 Protected a   
                 Infected b   
                 Ratio 
                 % 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 2773 
                 1 
                 0 
                 12 
                 0 
                 12/12 
                 100%  
               
               
                 2495 
                 1 
                 0 
                 11 
                 0 
                 11/11 
                 100%  
               
               
                 2075 
                 1 
                 0 
                 10 
                 0 
                 10/10 
                 100%  
               
               
                 2004 
                 1 
                 0 
                 9 
                 0 
                 9/9 
                 100%  
               
               
                 633 
                 1 
                 0 
                 8 
                 0 
                 8/8 
                 100%  
               
               
                 618 
                 0 
                 1 
                 7 
                 1 
                 7/8 
                 88% 
               
               
                 563 
                 1 
                 0 
                 7 
                 1 
                 7/8 
                 88% 
               
               
                 384 
                 1 
                 0 
                 6 
                 1 
                 6/7 
                 86% 
               
               
                 372 
                 1 
                 0 
                 5 
                 1 
                 5/6 
                 83% 
               
               
                 272 
                 1 
                 0 
                 4 
                 1 
                 4/5 
                 80% 
               
               
                 176 
                 0 
                 1 
                 3 
                 2 
                 3/5 
                 60% 
               
               
                 162 
                 1 
                 0 
                 3 
                 2 
                 3/5 
                 60% 
               
               
                 115 
                 1 
                 0 
                 2 
                 2 
                 2/4 
                 50% 
               
               
                 75 
                 0 
                 1 
                 1 
                 3 
                 1/4 
                 25% 
               
               
                 68 
                 0 
                 1 
                 1 
                 4 
                 1/5 
                 20% 
               
               
                 65 
                 0 
                 1 
                 1 
                 5 
                 1/6 
                 17% 
               
               
                 56 
                 0 
                 1 
                 1 
                 6 
                 1/7 
                 14% 
               
               
                 55 
                 1 
                 0 
                 1 
                 6 
                 1/7 
                 14% 
               
               
                 55 
                 0 
                 1 
                 0 
                 7 
                 0/7 
                  0% 
               
               
                 53 
                 0 
                 1 
                 0 
                 8 
                 0/8 
                  0% 
               
               
                 27 
                 0 
                 1 
                 0 
                 9 
                 0/9 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 10 
                  0/10 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 11 
                  0/11 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 12 
                  0/12 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 13 
                  0/13 
                  0% 
               
               
                 17 
                 0 
                 1 
                 0 
                 14 
                  0/14 
                  0% 
               
               
                   
               
               
                   a Sum from the bottom. 
               
               
                   b Sum from the top 
               
               
                   c Endpoint protection titer (50% protective titer) was calculated to be 1:115 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 16 
               
             
            
               
                   
               
               
                 SHIVDH12-V3AD8 
               
            
           
           
               
               
               
               
            
               
                 Endpoint 
                   
                   
                   
               
               
                 neutral- 
                   
                   
                   
               
               
                 ization 
                   
                   
                   
               
               
                 titer in 
                 Animal number 
                 Accumulated value 
                 Protected 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 plasma 
                 Protected 
                 Infected 
                 Protected a   
                 Infected b   
                 Ratio 
                 % 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 13805 
                 1 
                 0 
                 16 
                 0 
                 16/16 
                 100%  
               
               
                 13120 
                 1 
                 0 
                 15 
                 0 
                 15/15 
                 100%  
               
               
                 12659 
                 1 
                 0 
                 14 
                 0 
                 14/14 
                 100%  
               
               
                 12055 
                 1 
                 0 
                 13 
                 0 
                 13/13 
                 100%  
               
               
                 2282 
                 1 
                 0 
                 12 
                 0 
                 12/12 
                 100%  
               
               
                 1972 
                 1 
                 0 
                 11 
                 0 
                 11/11 
                 100%  
               
               
                 539 
                 0 
                 1 
                 10 
                 1 
                 10/11 
                 91% 
               
               
                 422 
                 1 
                 0 
                 10 
                 1 
                 10/11 
                 91% 
               
               
                 420 
                 1 
                 0 
                 9 
                 1 
                  9/10 
                 90% 
               
               
                 376 
                 1 
                 0 
                 6 
                 1 
                 8/9 
                 89% 
               
               
                 159 
                 1 
                 0 
                 7 
                 1 
                 7/8 
                 85% 
               
               
                 143 
                 1 
                 0 
                 6 
                 1 
                 8/7 
                 86% 
               
               
                 142 
                 1 
                 0 
                 5 
                 1 
                 5/6 
                 83% 
               
               
                 136 
                 0 
                 1 
                 4 
                 2 
                 4/6 
                 67% 
               
               
                 106 
                 1 
                 0 
                 4 
                 2 
                 4/6 
                 67% 
               
               
                 101 
                 1 
                 0 
                 3 
                 2 
                 3/5 
                      60% c   
               
               
                 90 
                 0 
                 1 
                 2 
                 3 
                 2/5 
                      40% c   
               
               
                 80 
                 1 
                 0 
                 2 
                 3 
                 2/5 
                 40% 
               
               
                 70 
                 0 
                 1 
                 1 
                 4 
                 1/5 
                 20% 
               
               
                 52 
                 1 
                 0 
                 1 
                 4 
                 1/5 
                 20% 
               
               
                 39 
                 0 
                 1 
                 0 
                 5 
                 0/5 
                  0% 
               
               
                 35 
                 0 
                 1 
                 0 
                 6 
                 0/6 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 7 
                 0/7 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 8 
                 0/8 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 9 
                 0/9 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 10 
                  0/10 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 11 
                  0/11 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 12 
                  0/12 
                  0% 
               
               
                 20 
                 0 
                 1 
                 0 
                 13 
                  0/13 
                  0% 
               
               
                   
               
               
                   a Sum from the bottom. 
               
               
                   b Sum from the top. 
               
               
                   c Endpoint protection titer (50% protective titer) was calculated to be 1:95.5 
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
             
               
                 TABLE 17 
               
             
            
               
                   
               
               
                   
                 Weeks 
                 Pre-Infection 
                 Pre mAb Treatment 
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Post 
                 CD4 +T Cells 
                 CD4 +T cells 
                 Viral Load 
                 Clinical 
               
               
                 Animal 
                 infection 
                 cells/μl 
                 cells/μl 
                 RNA Copies/ml 
                 Status 
               
               
                   
               
               
                 DBZ3 
                 159 
                  650 
                 118 
                 1.08E+04 
                 Asymptomatic 
               
               
                 DC99A 
                 159 
                  623 
                 165 
                 7.60E+03 
                 Asymptomatic 
               
               
                 DBXE 
                 163 
                 1585 
                 158 
                 1.96E+05 
                 Intermittent diarrhea 
               
               
                 DCF1 
                 157 
                 1203 
                 105 
                 1.44E+05 
                 Intermittent diarrhea 
               
               
                 DCM8 
                 163 
                  608 
                  43 
                 1.59E+03 
                 Intermittent diarrhea 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
             
               
                   
                 TABLE 18 
               
               
                   
                   
               
               
                   
                   
                   
                 SIV Gag 
                 SW Gag 
               
               
                   
                   
                   
                 RNA 
                 DNA 
               
               
                   
                   
                 Treatment 
                 Copies 
                 Copies 
               
               
                   
                   
                 Time 
                 per 10 8   
                 per 10 8   
               
               
                   
                 Animal 
                 (Days) 
                 Cell Eq 
                 Cell Eq 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 DBZ3 
                  0 
                 9.000 
                 6.700 
               
               
                   
                 DBZ3 
                 10 
                 360 
                 7.600 
               
               
                   
                 DBZ3 
                 20 
                 2.400 
                 14.000 
               
               
                   
                 DC99A 
                  0 
                 31.000 
                 1.400 
               
               
                   
                 DC99A 
                 14 
                 18.000 
                 5.600 
               
               
                   
                 DC99A 
                 20 
                 8.100 
                 2.700 
               
               
                   
                 DBXE 
                  0 
                 470.000 
                 71.000 
               
               
                   
                 DBXE 
                 14 
                 17.000 
                 33.000 
               
               
                   
                 DBXE 
                 17 
                 11.000 
                 22.000 
               
               
                   
                 DCMB 
                  0 
                 110.000 
                 8.600 
               
               
                   
                 DCMB 
                 14 
                 1.700 
                 1.600 
               
               
                   
                 DCMB 
                 20 
                 22.000 
                 6.600 
               
               
                   
                 DCF1 
                  0 
                 240.000 
                 15.000 
               
               
                   
                 DCF1 
                 14 
                 190.000 
                 11.000 
               
               
                   
                 DCF1 
                 20 
                 1100.000 
                 14.000 
               
               
                   
                   
               
            
           
         
       
     
     The foregoing examples and description of the preferred embodiments should be taken as illustrating, rather than as limiting the present invention as defined by the claims. As will be readily appreciated, numerous variations and combinations of the features set forth above can be utilized without departing from the present invention as set forth in the claims. Such variations are not regarded as a departure from the scope of the invention, and all such variations are intended to be included within the scope of the following claims. All references cited herein are incorporated herein in their entireties.