EA042696B1 - HUMAN IMMUNODEFICIENCY VIRUS NEUTRALIZING ANTIBODIES AND METHODS FOR THEIR APPLICATION - Google Patents

Description

Cross-reference to related applications

This application claims priority under 35 U.S.C. §119(e) US Provisional Application No. 61/486960, filed May 17, 2011. Its disclosure is incorporated herein in its entirety.

Publicly funded research statement

The research that led to the present invention was supported in part by National Institutes of Health Grant No. P01 AI08677-01. Accordingly, the US government has certain rights to this invention.

Field of invention

The present invention relates to antibodies directed to human immunodeficiency virus (HIV) epitopes. The present invention further relates to the production and use of broad spectrum neutralizing antibodies directed to the HIV gp120 envelope protein for the prevention and treatment of HIV infection.

Background of the invention

HIV causes acquired immunodeficiency syndrome (AIDS). The immune response to HIV infection in nonprogressors indicates that specific viral immunity can limit infection and disease symptoms. In some HIV-infected individuals, broad-spectrum neutralizing IgG antibodies have been detected in serum; little is known about the specificity and activity of these antibodies, despite their possible importance in the development of effective vaccines, and no characteristic has been correlated with protective immunity. In animal models, passive transfer of neutralizing antibodies may contribute to protection against viral challenge. HIV-infected individuals can also develop neutralizing humoral responses, but the detailed composition of the serological response is still not fully known.

Many immunological pathologies have been described for AIDS. These include, but are not limited to, abnormal B cell function, abnormal humoral response, impaired monocyte cellular function, impaired cytokine production, suppressed natural killer and decreased cytotoxic cell function, impaired ability of lymphocytes to recognize and respond to soluble antigens, and population depletion. T4 lymphocytes helpers/inducers.

Amino acid and RNA sequences encoding the env of HIV are known from a variety of HIV strains (Modrow, S. et al., J. Virology, 61(2):570 (1987)). The HIV virion is coated with a membrane or envelope derived from the outer membrane of the host cells. This membrane contains a population of envelope glycoproteins (gp160) anchored in the membrane bilayer by a carboxy-terminal region. Each glycoprotein contains two segments: an N-terminal segment and a C-terminal segment. The N-terminal segment, called gp120 due to its relative molecular weight of approximately 120 kDa, protrudes into the aqueous environment surrounding the virion. The C-terminal segment, called gp41, spans the membrane. The N-terminal gp120 and C-terminal gp41 are covalently linked via a peptide bond, which is particularly susceptible to proteolytic cleavage. See European Patent Application Publication No. 0335635 by McCune et al. and the references cited therein, each of which is incorporated herein by reference in its entirety.

Several approaches have been proposed for the development of an AIDS vaccine, including, but not limited to, inactivated and attenuated antiviral vaccines, subunit vaccines from virus-infected cells, recombinant viral antigens, synthetic peptide vaccines, anti-idiotypic vaccines, and carrier-virus vaccines. . An additional approach to the therapeutic and prophylactic treatment of HIV involves the production of highly effective broad spectrum neutralizing monoclonal antibodies. Many studies have reported cloning and generating monoclonal antibodies in various ways to target the CD4 binding site as well as other parts of the virion spike and to neutralize HIV. In general, these methods include self-fusion or phage display methods. Typically, when generating neutralizing antibodies against HIV using phage display methods, random combinations of heavy and light chains are combined and a random pair is selected. Studies have reported a limited number of monoclonal antibodies, such as, for example, the b12 phage display antibody, that are highly effective, broad spectrum, and neutralizing (meaning antibodies that can neutralize multiple strains of HIV in sera) against HIV. Monoclonal antibody b12 is a broad spectrum neutralizing antibody that has been reported to prevent HIV infection in monkeys. Another broad spectrum neutralizing antibody includes 2G12, which has an uncharacteristic structure that has not yet been found in any other antibody, with three antigen binding sites.

VRC01 is a newly discovered broad spectrum neutralizing antibody that targets the CD4 binding site (CD4bs) on the HIV spike. VRC01 was isolated by purifying single B cells that bind to a soluble biotin-labeled stabilized gp120 core fragment with resurfaced HIV (X. Wu et al., Science 329, 856 (Aug 13, 2010)). Although successful, isolation was inefficient, and only three closely related HIV-binding antibodies were obtained from 25 million peripheral blood mononuclear cells in one individual. Like other anti-HIV antibodies produced by the single cell antigen capture method, the VRC01-3 antibody showed a very high level of somatic mutations, which are necessary for efficacy and broad spectrum. This high mutation rate is a likely hindrance to antibody cloning, as mutated sequences may no longer be complementary primers used for cloning.

Some studies have reported that certain patients produce anti-HIV antibodies that are neutralizing and broad spectrum. Studies have reported that antibodies can protect against initial HIV infection in passive transfer experiments in non-human primates and can modulate viral load during infection. See, for example, Mascola, 2000; Shibata, 1999; Veazey, 2003; Parren, 2001; Mascola, 1999; Trkola, 2005; Wei, 2003; Frost, 2005; Burton, 2004; Mascola, 2007; Karlsson Hedestam, 2008; McMichael, 2006; Zolla-Pazner, 2004.

Brief summary of the invention

In one embodiment, the present invention relates to broad spectrum neutralizing antibodies against HIV. In one embodiment, the present invention relates to an isolated anti-HIV antibody containing a heavy chain that contains the consensus amino acid sequence:

QXXLXQSGGXVKKPGXSVXVSCXASGYXXFXXYXIHWXRQAPGXGXXWVGXIXPRXGXXXXAXX

FQGRLSLTRDXXXXXXTXXXFMDLXGLRXDTAVYFCARXXXXXXXXXXXXXXXXXXDX (SEQ ID NO: 1), where X is any or no amino acid.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing a light chain that contains the consensus amino acid sequence:

EIXLTQSPXSLSXSXGEXXTISCXXXQXXXXXXXLXWYQQRXGXARPLLIXXXSXXXXGVPXRF

SGXXXGXXYXLXISXLXXDDXAXYFCXXYEXXXXXXX (SEQ ID NO: 2), where X is any amino acid or no amino acid.

In yet another embodiment, the present invention provides an isolated anti-HIV antibody comprising a heavy chain that contains a highly conserved consensus sequence and a light chain that contains a highly conserved consensus sequence. The present invention further relates to a method for preparing an isolated anti-HIV antibody comprising a heavy chain that contains a highly conserved consensus sequence and a light chain that contains a highly conserved consensus sequence.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising the heavy chain consensus sequence of SEQ ID NO: 1 and the light chain sequence of SEQ ID NO: 2. In a further embodiment, the present invention provides an isolated anti-HIV antibody comprising one or both heavy chain consensus sequences of SEQ ID NO: 1 and light chain sequences of SEQ ID NO: 2, or sequences that are at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 97%, or at least 98%, or at least 99% identical to them, provided that the antibody does not have the VRC01 amino acid sequence.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising one or both of the heavy chain consensus sequences of SEQ ID NO: 1 and the light chain of SEQ ID NO: 2, and wherein the antibody neutralizes the ZM53M.PB12 HIV virus at an IC ₅₀ concentration of less than 1.0 µg/ml, or HIV virus R1166.c1 with an IC ₅₀ concentration of less than 1.0 µg/ml, or DU172.17 with an IC ₅₀ concentration of less than 30 µg/ml. In another embodiment, the present invention provides an isolated anti-HIV antibody comprising one or both of the heavy chain consensus sequences of SEQ ID NO: 1 and the light chain of SEQ ID NO: 2, wherein the antibody neutralizes a VRC01-resistant HIV virus at an IC ₅₀ concentration of less than than 30 mcg/ml.

In another embodiment, the present invention provides an isolated anti-HIV antibody selected from the group consisting of 3BNC117, 3BNC60, 12A12, 12A21, NIH45-46, 8ANC131, 8ANC134, IB2530, INC9, and 8ANC196.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising heavy chain CDR1, CDR2 and CDR3 regions and light chain CDR1, CDR2 and CDR3 regions comprising the amino acid sequences of the respective regions of an anti-HIV antibody selected from the group consisting of 3BNC117, 3BNC60 , 12A12, 12A21, NIH45-46, bANC131, 8ANC134, IB2530, INC9 and 8ANC196.

In another embodiment, the present invention relates to an isolated anti-HIV antibody comprising a heavy chain which contains an amino acid sequence selected from the group consisting of SEQ ID NO: 5-438.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing a light chain that contains an amino acid sequence selected from the group consisting of SEQ ID NO: 439-583.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing a heavy chain and a light chain, which contain the amino acid sequence set forth in table. A or tab. IN.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising an insert sequence comprising the amino acid sequence: ASWDFDF (SEQ ID NO: 3).

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising an insertion sequence comprising the amino acid sequence: TARDY (SEQ ID NO: 4).

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising the insertion sequences of SEQ ID NO: 3 and SEQ ID NO: 4.

In another embodiment, the present invention relates to a method for improving the HIV neutralization potency and scope of an isolated anti-HIV antibody, comprising inserting at least one insertion sequence of SEQ ID NO: 3 and SEQ ID NO: 4.

According to another embodiment, the present invention relates to compositions containing an isolated anti-HIV antibody of the invention.

In another embodiment, the present invention relates to pharmaceutical compositions comprising an antibody of the invention and a pharmaceutically acceptable carrier.

According to another embodiment, the present invention relates to nucleic acid molecules encoding an isolated anti-HIV antibody of the invention.

In other embodiments, the present invention relates to vectors containing nucleic acid molecules encoding an isolated anti-HIV antibody of the invention, and cells containing such vectors.

According to another embodiment, the present invention relates to a method for preventing or treating an HIV infection or an HIV-related disease, comprising the steps of: identifying a mammal in need of such prevention or treatment, and administering to said mammal a therapeutically effective amount of at least one anti-HIV antibody according to invention.

In another embodiment, the method further comprises administering a second therapeutic agent. According to another embodiment, the second therapeutic agent is an antiviral agent.

Another embodiment of the present invention relates to a method of reducing virus replication or spread of infection to additional host cells or tissues, which comprises contacting a mammalian cell with at least one antibody of the invention. In another aspect, the present invention relates to a method for treating a mammal infected with HIV, the method comprising administering to said mammal a pharmaceutical composition comprising at least one antibody of the invention.

According to another embodiment, the present invention relates to a method for preparing and administering an anti-HIV antibody preparation that is suitable for administration to a mammal that has or is at risk of HIV infection in an amount and in a schedule sufficient to induce a protective immune response against HIV, or reduction of the HIV virus in a mammal. In another embodiment, the present invention relates to a method for detecting an anti-HIV antibody containing a heavy chain that contains a highly conserved consensus sequence and a light chain that contains a highly conserved consensus sequence in a biological sample.

In another embodiment, the present invention provides isolated antibodies of the invention for use in the treatment of HIV.

In another embodiment, the present invention relates to a kit containing a pharmaceutically acceptable unit dose of a pharmaceutically effective amount of an isolated anti-HIV antibody of the invention and a pharmaceutically acceptable unit dose of a pharmaceutically effective amount of an anti-HIV agent selected from the group consisting of a non-nucleoside reverse transcriptase inhibitor, a protease inhibitor, an entry or fusion inhibitor; and integrase inhibitors, wherein the two pharmaceutically acceptable dosage units may optionally be in the form of a common pharmaceutically acceptable dosage unit.

In another embodiment, the present invention relates to a kit for diagnosing, predicting, or monitoring HIV treatment in an individual, which contains one or more detection reagents that specifically bind to neutralizing anti-HIV antibodies in a biological sample of the individual. In another aspect of the invention, the kit additionally provides reagents for performing PCR or mass spectrometry.

Brief description of the drawings

In FIG. 1A-D shows the neutralizing activity of an anti-HIV antibody at IC ₅₀ . (A) Limited panel. The upper line indicates the donor number, then the clone or antibody (Table 4); viruses are shown on the left. Colors indicate concentrations at IC ₅₀ : red <0.1 μg/ml; orange 0.1-1 µg/ml; yellow 1-10 mcg/ml; green >10 µg/mL; white - no neutralization at any tested concentration. (B) Extended panel. (C) Combined neutralization comparison plot for VRC01, NIH45-46, 3BNC117. The length of the lines and the size of the circles are inversely proportional to IC ₅₀ . Colors indicate viral clades: red A; blue B; green C; fuchsia D; black AE; golden AG. (D) Heavy chain sequence of 3BNC60, 1B2530 and 8ANC134 coated with peptides found by mass spectrometry in light gray. Red dots indicate differences from the corresponding germline sequences.

In FIG. 2A-C show the binding properties of anti-HIV antibodies. (A) Representative SPR sensorgrams for YU2-gp140 and 2CC-core binding by germline reversed antibodies 12A12, 12A21 and 12A (GL). (B) Graph shows K _A for representative antibodies. (C) Graph showing the mean fluorescence intensity of anti-CD4i antibody binding to Bal.26 expressing 293T cells after incubation with the labeled antibodies. The table indicates whether the antibody induces CD4i site accessibility or not.

In FIG. 3A and 3B show the consensus sequence of the anti-HIV antibody and the amino acid sequences of the anti-HIV antibodies. (A) Alignment of amino acids relative to the frame (FR) and CDR regions for consensus, germline genes, 10 selected antibodies and 8ANC195. Residues are numbered according to the 3BNC60 structure. (B) As in (A), for light chains. (C, D and E) Crystal structure of Fab 3BNC60.

In FIG. 4A and 4B show the yield of highly mutated immunoglobulin heavy chains using specific primers. (A) Joint comparison of the new and old primer set. Red cells indicate successful amplification of the IgV _H genes. (B) Anti-HIV antibodies that bind to the 2CC core of Pt 8. The clonal families are represented by differently unfolded sectors. The two high mutation clones that were not amplified with the old primer set are shown in the shaded sectors of the pie chart.

In FIG. 5 shows the heavy (A) and light (B) sequences of Ig V new clonal members of VRC01.

In FIG. 6 shows the neutralizing activity of the patient's serum. (A) The table summarizes the neutralizing activity of purified serum IgG against a panel of Tier 2 viruses in the Tzm-bl assay. Dark red cells show IC ₅₀ values below 10 µg/ml, orange between 10 and 100 µg/ml and yellow above 100 µg/ml. (B) Dot plot summarizes the IC50 values presented in A for 4 more comprehensively tested patients.

In FIG. 7 demonstrates the detection of antibodies by mass spectrometry. MS/MS spectrum of collision-activated dissociation recorded on the doubly charged peptides HSDYCDFDVWGSGSQVIVSSASTK from 3BNC153HC (A) and DGLGEVAPAYLYGIDAWGQGTTVIVTSASTK from 8ANC134HC. (B) Observed b-type fragment ions (containing the N-terminus) and y-type fragment ions (containing the C-terminus) are labeled on the spectrum. Water loss from fragment ions is indicated by *. Ions corresponding to the loss of water from the parent ion are labeled in the spectrum. The observed core splittings are denoted in the sequence 1 for b-type ions and L for y-type ions.

In FIG. 8A and 8B show the affinity of antibodies against HIV. (A) Antibody binding to gp140 and 2CC core as measured by surface plasmon resonance (SPR). The SPR sensorgrams for antibody binding selected clones of the 3BNC antibody are shown as a function of time. (B) The bar graph shows the binding affinity (K _A ) for gp140 and 2CC-core antigens for selected IgG antibodies shown in A. RU, response units.

In FIG. 9A-9C illustrate an analysis of somatic hypermutations of selected anti-HIV antibodies for the sequences of (A) the immunoglobulin heavy chain gene, (B) the anti-light chain gene, and (C) the light chain λ gene. The sequences are aligned with their respective germline nucleotide sequences. Somatic mutations are shown in red letters, additional gray boxes indicate substitutional mutations. The germline amino acid sequences, with consensus residues denoting, are shown above the nucleotide alignment.

In FIG. 10A-10C show the antibody sequences from one propagated neutralizing clone in each patient (A) (Pt)1, (B) Pt3, and (C) Pt8. Peptides identified by mass spectrometry are indicated by color. Variants marked with an asterisk are uniquely identifiable by one or more mass spectrometrically observable peptides (shown in light grey). The rest of the mass spectrometrically observed peptides are mapped non-uniquely in a variety of ways, as shown in dark gray. Underlined amino acids denote non-trypsinolytic cleavage sites in the variants shown. The cleavage is thought to occur via chymotrypsinolytic cleavage or additional mutations (not seen among cloned variants) that place a lysine or arginine residue at these sites.

Detailed description of the invention

I. Antibodies that neutralize HIV.

In one embodiment, the present invention relates to broad spectrum neutralizing antibodies against HIV. In one embodiment, the present invention relates to an isolated anti-HIV antibody containing a heavy chain that contains the consensus amino acid sequence:

QXXLXQSGGXVKKPGXSVXVSCXASGYXXFXXYXIHWXRQAPGXGXXWVGXIXPRXGXXXXAXX

FQGRLSLTRDXXXXXXTXXXFMDLXGLRXDTAVYFCARXXXXXXXXXXXXXXXXXXDX (SEQ ID NO: 1), where X is any or no amino acid.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing a light chain that contains the consensus amino acid sequence:

EIXLTQSPXSLSXSXGEXXTISCXXXQXXXXXXXLXWYQQRXGXARPLLIXXXSXXXXGVPXRF

SGXXXGXXYXLXISXLXXDDXAXYFCXXYEXXXXXXX (SEQ ID NO: 2), where X is any amino acid or no amino acid.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising the heavy chain sequence of SEQ ID NO: 1 and the light chain sequence of SEQ ID NO: 2. In a further embodiment, the present invention provides an isolated anti-HIV antibody comprising one or both of the sequences heavy chain of SEQ ID NO: 1 and light chain of SEQ ID NO: 2, or sequences that are at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 97%, or at least 98%, or at least 99% identical with them, provided that the antibody does not have the VRC01 amino acid sequence. Percent identity is determined as described later in this document.

The present invention in other embodiments provides an isolated anti-HIV antibody comprising a heavy chain that contains a highly conserved heavy chain amino acid sequence and a light chain that contains a highly conserved light chain amino acid sequence. A highly conserved heavy chain amino acid sequence is defined herein as an amino acid sequence that is at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 97%, or at least 98%, or at least 99% identical to the sequence of SEQ ID NO: 1. A highly conserved light chain amino acid sequence is defined herein as an amino acid sequence that at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 97%, or at least 98%, or at least 99% identical to the sequence of SEQ ID NO: 2. Percent identity is determined as described later in this document.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising a heavy chain that contains a highly conserved heavy chain amino acid sequence and a light chain that contains a highly conserved light chain amino acid sequence, provided that the antibody does not have a VRC01 sequence.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing one or both of the heavy chain sequences of SEQ ID NO: 1 and the light chain of SEQ ID NO: 2, de the antibody neutralizes the ZM53M.PB12 HIV virus at an IC ₅₀ concentration of less than 1, 0 µg/ml, or HIV virus R1166.c1 with an IC ₅₀ concentration of less than 1.0 µg/ml, or DU172.17 with an IC ₅₀ concentration of less than 30 µg/ml. In another embodiment, the present invention relates to an isolated anti-HIV antibody comprising one or both of the heavy chain sequences of SEQ ID NO: 1 and the light chain of SEQ ID NO: 2, wherein the antibody neutralizes a VRC01-resistant HIV virus at an IC ₅₀ concentration of less than 30 µg/ml. A VRC01-resistant HIV virus is defined herein as an HIV virus that is resistant to VRC01 neutralization with an IC ₅₀ value of 5 0 μg/mL. HIV viruses resistant to VRC01 include, for example, HO86.8, DU172.17, 250-4, 278-50 and 620345.C1.

In another embodiment, the present invention provides an isolated anti-HIV antibody selected from the group consisting of 3BNC117, 3BNC60, 12A12, 12A21, NIH45-46, bANC131, 8ANC134, IB2530, INC9, and 8ANC196.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising heavy chain CDR1, CDR2 and CDR3 regions and light chain CDR1, CDR2 and CDR3 regions containing the amino acid sequences of the corresponding regions of the anti-HIV antibody.

HIV selected from the group consisting of 3BNC117, 3BNC60, 12A12, 12A21, NIH45-46, bANC131,

8ANC134, IB2530, INC9 and 8ANC196.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing a heavy chain that contains an amino acid sequence selected from the group consisting of SEQ ID NO: 5-438.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing a light chain that contains an amino acid sequence selected from the group consisting of SEQ ID NO: 439-583.

In another embodiment, the present invention relates to an isolated anti-HIV antibody containing a heavy chain and a light chain, which contains the amino acid sequence set forth in table. A or B.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising an insert sequence comprising the amino acid sequence: ASWDFDF (SEQ ID NO: 3). In a further embodiment, the present invention relates to an isolated anti-HIV antibody, wherein the insertion sequence of SEQ ID NO: 3, which corresponds to the heavy chain FR3 region starting at amino acid 74 in 3BNC117 and 3BNC60, as shown in FIG. 5A replace the corresponding region determined by sequence alignment with an anti-HIV antibody of the invention. For example, SEQ ID NO: 3 may be inserted after the seventh amino acid of heavy chain FR3.

In another embodiment, the present invention provides an isolated anti-HIV antibody comprising an insertion sequence comprising the amino acid sequence: TARDY (SEQ ID NO: 4). In a further embodiment, the present invention relates to an isolated anti-HIV antibody, wherein the insertion sequence of SEQ ID NO: 4, which corresponds to the heavy chain CDR3 region starting at amino acid 103 in NIH45-46, as shown in FIG. 5A replace the corresponding region determined by sequence alignment with an anti-HIV antibody of the invention. For example, SEQ ID NO: 4 may be inserted after the fourth amino acid in the heavy chain CDR3.

In another embodiment, the present invention relates to an isolated anti-HIV antibody, wherein the insertion sequence of SEQ ID NO: 3, which corresponds to the heavy chain FR3 region starting at amino acid 74 in 3BNC117 and 3BNC60, as shown in FIG. 5A, replace the corresponding region, determined by sequence alignment, of an anti-HIV antibody of the invention with the insertion sequence of SEQ ID NO: 4, which corresponds to the heavy chain CDR3 region starting at amino acid 103 in NIH45-46, as shown in FIG. 5A replace the corresponding region determined by sequence alignment with an anti-HIV antibody of the invention. For example, SEQ ID NO: 3 may be inserted after the seventh amino acid of the heavy chain FR3 and SEQ ID NO: 4 may be inserted after the fourth amino acid of the heavy chain CDR3.

In a further embodiment, the present invention relates to a method for improving the HIV neutralization efficiency and width of an isolated anti-HIV antibody, comprising obtaining an isolated anti-HIV antibody, wherein the insertion sequence is SEQ ID NO: 3, which corresponds to the heavy chain FR3 region starting at amino acid 74 in 3BNC117 and 3BNC60 as shown in Fig. 5A, replace the corresponding region determined by sequence alignment with an anti-HIV antibody of the invention and/or with an insertion sequence of SEQ ID NO: 4 that corresponds to the heavy chain CDR3 region starting at amino acid 103 in NIH45-46 as shown in FIG. 5A replace the corresponding region determined by sequence alignment with an anti-HIV antibody of the invention. For example, SEQ ID NO: 3 may be inserted after the seventh amino acid of the heavy chain FR3 and/or SEQ ID NO: 4 may be inserted after the fourth amino acid of the heavy chain CDR3. One skilled in the art can modify the amino acid sequence of an antibody using recombinant techniques and/or synthetic chemical techniques to produce a polypeptide or antibody. Also, one skilled in the art can identify an improved anti-HIV antibody with higher potency and neutralization width by using an HIV neutralization assay, as described below.

In another embodiment, the present invention relates to an improved isolated anti-HIV antibody comprising at least one of the insertion sequences of SEQ ID NO: 3 and SEQ ID NO: 4, wherein the improved isolated anti-HIV antibody has higher HIV neutralization efficiency and width than said isolated anti-HIV antibody without the insertion sequences of SEQ ID NO: 3 and SEQ ID NO: 4. One skilled in the art can identify an improved anti-HIV antibody with higher HIV neutralization potency and breadth by using an HIV neutralization assay as described below.

- 6 042696

One skilled in the art can modify the amino acid sequence of an antibody using recombinant techniques and/or synthetic chemical techniques to produce a polypeptide or antibody.

In another embodiment, the present invention relates to a method for producing an isolated anti-HIV antibody comprising the heavy chain consensus sequence of SEQ ID NO: 1 and the light chain of SEQ ID NO: 2. In a further embodiment, the present invention relates to a method for producing an isolated anti-HIV antibody comprising one or both of the heavy chain consensus sequence of SEQ ID NO: 1 and the light chain of SEQ ID NO: 2, or sequences that are at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 97%, or at least 98%, or at least 99% identical with them, provided that the antibody does not have an amino acid sequence VRC01. Percent identity is determined as described later in this document.

In another embodiment, the present invention relates to a method for detecting an isolated anti-HIV antibody, comprising obtaining an immunoglobulin-containing biological sample from a mammal, isolating the anti-HIV antibody from said sample, determining the amino acid sequence of the anti-HIV antibody, and identifying the presence of the heavy chain sequence of SEQ ID NO: 1, and light chain SEQ ID NO: 2. In a further embodiment, the present invention relates to a method for selecting an isolated anti-HIV antibody, comprising determining the presence of one or both of the heavy chain consensus sequence of SEQ ID NO: 1 and the light chain of SEQ ID NO: 2, or sequences which are at least 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 97%, or at least 98%, or at least 99% identical to them, provided that the antibody does not have the VRC01 amino acid sequence. Percent identity is determined as described later in this document. The biological sample may be blood, serum, saliva, urine, sputum, a cell scraping sample, or a tissue biopsy. Amino acid sequences can be determined by methods known in the art, including, for example, PCR and mass spectrometry.

The term antibody (Ab) as used herein includes monoclonal antibodies, polyclonal antibodies, polyspecific antibodies (eg, bispecific antibodies and polyreactive antibodies), and antibody fragments. Thus, the term antibody, as used in any context herein, includes, but is not limited to, any specific binding element, immunoglobulin class, and/or isotype (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA, IgD, IgE, and IgM) and a biologically significant fragment or specific binding element 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 that contains at least two heavy (H) chains and two light (L) chains mutually linked via disulfide bonds, or an antigen-binding portion thereof. The heavy chain consists of the heavy chain variable region (VH) and the heavy chain constant region (CH1, CH2 and CH3). The light chain consists of a light chain variable region (V _L ) and a light chain constant region (C _L ).

The variable regions of both heavy and light chains contain framework regions (FWRs) and complementarity determining regions (CDRs). The four FWR regions are relatively conserved, while the CDR regions (CDR1, CDR2 and CDR3) represent hypervariable regions and are arranged from the NH ₂ terminus to the COOH terminus as follows: FWR1, CDR1, FWR2, CDR2, FWR3, CDR3, FWR4. The variable regions of the heavy and light chains contain a binding domain that interacts with the antigen, while, depending on 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, and antibodies selected from libraries using enrichment methods available to those of ordinary skill in the art.

The term variable refers to the fact that, among antibodies, certain segments in the (V) domains vary greatly in sequence. The V domain mediates antigen binding and determines the specificity of a particular antibody for its particular antigen. However, the variability is unevenly distributed across the 110-amino acid stretch of the variable regions. In contrast, V regions are composed of relatively invariant fragments called framework regions (FRs) of 15-30 amino acids, which are separated by shorter regions of marginal variability called hypervariable regions, each 9-12 amino acids in length. The variable regions of the native heavy and light chains each contain four FRs, mostly taking on the beta sheet configuration, which are connected via hypervariable regions that form loops connecting the beta sheet structure, and in some cases forming part of it. The hypervariable regions in each chain are held together in

- 7 042696 close proximity due to FR and together with hypervariable regions from another chain contribute to the formation of the antigen-binding site of the antibody (see, for example, Rabat 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 contains amino acid residues from a complementarity determining region (CDR).

The term monoclonal antibody, as used herein, refers to an antibody derived from a population of substantially homogeneous antibodies, ie. individual antibodies contained in a population are identical except for possible naturally occurring mutations that may be present in minor amounts.

The term polyclonal antibody refers to preparations that contain different antibodies directed to different determinants (epitopes).

Monoclonal antibodies as used herein include chimeric antibodies in which a portion of the heavy and/or light chain is identical or homologous to corresponding sequences in antibodies derived from a particular species, or belongs to a particular class or subclass of antibodies, where the remainder of the chain(s) is identical or homologous corresponding sequences in antibodies derived from other species, or belong to a different class or subclass of antibodies, as well as fragments of such antibodies, provided that they exhibit the desired biological activity (see, for example, US patent No. 4816567; and Morrison et al., Proc Natl Acad Sci USA 81:6851-6855 (1984)). The present invention relates to antigen-binding variable region sequences derived from human antibodies. Accordingly, chimeric antibodies of primary interest herein include antibodies that have one or more human antigen-binding sequences (e.g., CDRs) and contain one or more sequences derived from an antibody that is not a human antibody, e.g., the FR region sequence or C. In addition, chimeric antibodies included herein are those that comprise a human variable region antigen-binding sequence of one antibody class or subclass and another sequence, such as an FR or C region sequence derived from another class or subclass of antibodies.

A humanized antibody is generally considered to be a human antibody that has one or more amino acid residues introduced into it from a non-human source. These non-human amino acid residues are often referred to as import residues, which are typically taken from the import variable region. Humanization can be carried out according to the method of Winter and colleagues (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 replacing the imported hypervariable region sequences with the corresponding human antibody sequences. Accordingly, such humanized antibodies are chimeric antibodies (see, for example, US Pat. No. 4,816,567) wherein significantly less than an intact human variable region is replaced with the corresponding sequence from a non-human species.

An antibody fragment contains a portion of an intact antibody, such as the antigen-binding or variable region of an intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab') ₂ and Fv fragments; diabody; linear antibodies (see, for example, US patent No. 5641870; Zapata et al., Protein Eng. 8(10): 1057-1062 [1995]); single chain antibody molecules; and polyspecific antibodies formed from antibody fragments.

Fv is the minimum antibody fragment that contains the complete antigen recognition and antigen binding site. This fragment contains a dimer of the variable region domain of one heavy and one light chain, which have a tight non-covalent bond. From the folding of these two domains emerge six hypervariable loops (three loops from each of the H and L chains) that provide amino acid residues for antigen binding and confer antigen binding specificity on the antibody. However, even one variable region (or half of the Fv containing only three antigen-specific CDRs) has the ability to recognize and bind an antigen despite having lower affinity than the entire binding site.

Single chain Fvs (sFv or scFv) are antibody fragments that contain the V _H and V _L domains of an antibody linked into a single polypeptide chain. The sFv polypeptide may further comprise a polypeptide linker between the VH and VL domains that allows 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, p. 269-315 (1994); Borrebaeck 1995, below.

The term diabody refers to small antibody fragments obtained by constructing sFv fragments with short linkers (approximately 5-10 residues) between the V _H and V _L domains so that interchain but not intrachain V-domain pairing is achieved, leading to bivalent 042696 fragment, i.e. a fragment that has two antigen binding sites. Bispecific diabodies are heterodimers of two sFv crossover fragments in which the VH and _VL domains of the two antibodies are present in different polypeptide chains. Diabodies are described more fully, for example, in EP 404097; WO 93/11161 and Hollinger et al., Proc. Natl. Acad. sci. USA 90:6444-6448 (1993).

Domain antibodies (dAbs), which can be made in fully humanized form, are the smallest known antigen-binding antibody fragments ranging from about 11 to about 15 kDa. dAbs are functional variable regions of immunoglobulin heavy and light chains (VH and V _L , respectively). They are highly expressed in microbial cell culture, show favorable biophysical properties including, but not limited to, solubility and temperature stability, and are well suited for selection and affinity maturation through in vitro selection systems such as, for example, phage display. . dAbs are biologically active as monomers and, due to their small size and inherent stability, they can be formatted into larger molecules in order to create drugs with prolonged serum half-life or other pharmacological activities. Examples of this technique are described, for example, in WO 9425591 for antibodies derived from the heavy chain of Ig Camelidae, as well as in US 20030130496, which describes the isolation of single-domain fully human antibodies from phage libraries.

Fv and sFv are only particles with intact antigen binding sites that lack constant regions. Thus, they are suitable for reduced non-specific binding during in vivo use. sFv fusion proteins can be designed to achieve fusion of the effector protein at the amino or carboxy terminus of the sFv. See, for example, Antibody Engineering, ed. Borrebaeck, supra. An antibody fragment can also be a linear antibody, for example, as described in, for example, US Pat. No. 5,641,870. Such linear antibody fragments can be monospecific or bispecific.

In certain embodiments, the antibodies of the invention described are bispecific or multispecific. Bispecific antibodies are antibodies that have binding specificities for at least two different epitopes. Exemplary bispecific antibodies can bind two different epitopes on the same antigen. Other such antibodies may combine the first antigen-binding site with the binding site for the second antigen. Alternatively, the anti-HIV fragment can be combined with a Fab fragment that binds to a trigger molecule on the leukocyte, such as a T cell receptor molecule (for example, CD3), or an Fc receptor for IgG (FcyR), such as FcyRI (CD64), (CD32) and FcyRIII (CD16) in order to focus and localize cellular defense mechanisms on infected cells. Bispecific antibodies can also be used to localize cytotoxic agents to infected cells. Bispecific antibodies can be prepared as full length antibodies or antibody fragments (eg, F(ab') ₂ bispecific antibodies). For example, WO 96/16673 describes an anti-ErbB2/anti-FcyRIII bispecific antibody and US Pat. No. 5,837,234 discloses an anti-ErbB2/anti-FcyRI bispecific antibody. For example, a bispecific antibody against ErbB2/Fca has been reported in WO 98/02463; US Pat. No. 5,821,337 describes an anti-ErbB2/anti-CD3 bispecific antibody. See also, for example, Mouquet et al., Polyreactivity Increases The Apparent Affinity Of Anti-HIV Antibodies By Heteroligation. Nature, 467, 591-5 (2010).

Methods for producing bispecific antibodies are known in the art. The 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 techniques are disclosed, for example, in WO 93/08829, Traunecker et al., EMBO J., 10:3655-3659 (1991) and see also; Mouquet et al., Polyreactivity Increases The Apparent Affinity Of Anti-HIV Antibodies By Heteroligation. Nature, 467, 591-5 (2010).

Alternatively, antibody variable regions with desired binding specificities (antibody-antigen binding regions) are fused to immunoglobulin constant domain sequences. The fusion occurs using an Ig heavy chain constant domain containing at least a portion of the hinge region and the CH2 and CH3 regions. In some embodiments, the first heavy chain constant region (CH1) containing the site required for binding to the light chain is present in at least one of the fusion proteins. DNA encoding immunoglobulin heavy chain fusion proteins and, optionally, immunoglobulin light chain, are inserted into separate expression vectors and co-transfected into a suitable host cell. This provides greater flexibility in adjusting the mutual proportions of the three polypeptide fragments in embodiments where unequal ratios of the three polypeptide chains used in the construct provide optimal yield of the desired bispecific antibody. However, it is possible to insert coding sequences for two or all three polypeptide chains into one expression vector when expression of at least two polypeptide chains in equal ratios results in high yield or when the ratios do not significantly affect the yield of the desired combination of chains.

Methods for generating bispecific antibodies from antibody fragments are also described in the literature. For example, bispecific antibodies can be made using chemical linkage. For example, Brennan et al., Science, 229:81 (1985) describe a procedure where 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 the vicinal dithiols and prevent the formation of intermolecular disulfides. The resulting Fab' fragments are then converted into thionitrobenzoate (TNB) derivatives. Then one of the Fab'-TNB derivatives is reconverted to Fab'-thiol by reduction with mercaptoethylamine and mixed with an equimolar amount of another Fab'-TNB derivative to form a bispecific antibody. The resulting bispecific antibodies can be used as agents for selective immobilization of enzymes.

Other modifications of the antibody are provided herein. For example, the antibody can be crosslinked with one of a variety of non-protein polymers, such as polyethylene glycol, polypropylene glycol, polyoxyalkylenes, or copolymers of polyethylene glycol and polypropylene glycol. The antibody can also be encapsulated, for example, by coacervation or interphase polymerization methods (e.g., hydroxymethylcellulose or gelatin microcapsules and polymethyl methacrylate microcapsules, respectively), colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. Such methods are disclosed, for example, in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980).

Typically, the antibodies of the invention are produced by recombinant methods, using vectors and methods available in the art. Human antibodies can also be generated using B cells activated in vitro (see, for example, US Pat. Nos. 5,567,610 and 5,229,275). General methods in molecular genetics and genetic engineering that can be used 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, CA), 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, CA), Culture of Animal Cells: A Manual of Basic Technique, 2nd Ed. (R.I. Freshney. 1987. Liss, Inc. New York, NY) and Gene Transfer and Expression Protocols, pp. 109-128, ed. E.J. Murray, The Humana Press Inc., Clifton, N.J.). Reagents, cloning vectors, and genetic manipulation kits are available from commercial vendors such as BioRad, Stratagene, Invitrogen, ClonTech, and Sigma-Aldrich Co.

Human antibodies can also be generated in transgenic animals (eg mice) that are capable of producing the full repertoire of human antibodies in the absence of endogenous immunoglobulin production. For example, homozygous deletion of the antibody heavy chain junction region (JH) gene in chimeric and germline mutant mice has been described to result in complete inhibition of endogenous antibody production. Transfer of the human germline immunoglobulin gene array into such germline 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); US patent No. 5545806, 5569825, 5591669 (all GenPharm); US patent No. 5545807; and WO 97/17852. Such animals can be genetically engineered to produce human antibodies containing a polypeptide of the invention.

Various methods have been developed for obtaining antibody fragments. Traditionally, these fragments have been prepared by proteolytic cleavage of intact antibodies (see, e.g., 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 obtained directly from recombinant host cells. E. coli can express and secrete fragments of Fab, Fv and ScFv antibodies, thus making it possible to easily obtain large amounts of these fragments. Fab'-SH fragments can be directly isolated from E. coli and chemically linked to form F(ab') ₂ fragments (see, for example, Carter et al., Bio/Technology, 10:163-167 (1992) ). According to another approach, fragments of F(ab') ₂ can be isolated directly from the culture of recombinant host cells. Fab and F(ab')2 fragments with increased in vivo half-life containing rescue receptor binding epitope residues are described in US Pat. No. 5,869,046.

Other methods known in the art for selecting antibody fragments from libraries using enrichment techniques include, but are 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), can be used as an alternative previously reviewed

- 10 042696 new methods for selecting single chain antibodies. Single chain antibodies are selected from a library of single chain antibodies generated directly using filamentous phage techniques. The phage display technique is known in the art (eg, see method from Cambridge Antibody Technology (CAT)), as disclosed in US Pat. Nos. 5,565,332; 5733743; 5871907; 5872215; 5885793; 5962255; 6140471; 6225447; 6291650; 6492160; 6521404; 6544731; 6555313; 6582915; 6593081, as well as other members of the US family, or applications that rely on the priority filing of GB 9206318, which was filed May 24, 1992; see also Vaughn, et al. 1996, Nature Biotechnology, 14:309-314). Single chain antibodies can also be designed and constructed using an available recombinant DNA technique, such as a DNA amplification technique (eg, PCR), or possibly by using an appropriate hybridoma cDNA as a template.

Variant antibodies are also included within the scope of the invention. Thus, variants of the sequences listed in the invention are also included in the scope of the invention. Additional antibody sequence variants having improved affinity can be generated using methods known in the art and are included within the scope of the invention. For example, amino acid substitutions can be used to generate antibodies with further improved affinity. Alternatively, codon optimization of a nucleotide sequence can be used to improve translation efficiency in expression systems for antibody production.

Such variant antibody sequences will have 70% or more (ie 80%, 85%, 90%, 95%, 97%, 98%, 99% or more) sequence identity with the sequences listed herein. This sequence identity is calculated relative to the total length of the reference sequence (ie, the sequence specified in the description). Percent identity, as specified herein, is what is determined using BLAST version 2.1.3 using default parameters that are well defined in NCBI (National Center for Biotechnology Information; http://www.ncbi.nlm. nih.gov/) [Blosum Matrix 62; gap opening penalty = 11 and gap extension penalty = 1]. For example, the present invention relates to peptide sequences that contain at least about 5, 10, 15, 20, 30, 40, 50, 75, 100, 150 or more overlapping peptides from one or more of the sequences described herein, and also all intermediate lengths between these values. As used herein, the term intermediate lengths is intended to describe any length between the specified 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 relates to antibodies, either alone or in combination with other antibodies, such as, but not limited to, VRC01 and PG9, which have broad spectrum neutralizing activity in serum.

According to another embodiment, the present invention relates to methods for preparing and administering an anti-HIV antibody composition that is suitable for administration to a human or non-human primate patient who has HIV infection or is at risk of HIV infection, in an amount and according to a schedule sufficient in order to induce a protective immune response against HIV or a decrease in the HIV virus in a person.

According to another embodiment, the present invention relates to a vaccine comprising at least one antibody of the invention and a pharmaceutically acceptable carrier. In one embodiment, the vaccine is a vaccine that contains at least one antibody described herein and a pharmaceutically acceptable carrier. The vaccine may contain a plurality of antibodies having the characteristics described herein, in any combination, and, in addition, may contain antibodies that neutralize HIV, as is known in this field.

It should be understood that the compositions may be single or combinations of the antibodies described herein, which may be the same or different, to prophylactically or therapeutically treat the progression of various subtypes of HIV infection following vaccination. Such combinations can be selected according to the desired immunity. When the antibody is administered to an animal or human, it can be combined with one or more pharmaceutically acceptable carriers, excipients, or adjuvants that are known to those of ordinary skill in the art. The composition may additionally contain broad spectrum neutralizing antibodies known in the art, including but not limited to VRC01, PG9, and bl2.

In addition, with regard to determining the effective level in a patient for treating HIV, in particular, suitable animal models are available and widely used to evaluate the effectiveness of various anti-HIV gene therapy protocols in vivo (Sarver et al. (1993b), supra). These models include mice, monkeys and cats. Even though these animals are not naturally susceptible to HIV disease, chimeric mouse models (e.g., SCID, bg/nu/xid, NOD/SCID, SCID-hu,

- 11 042696 immunocompetent SCID-hu, BALB/c with bone marrow removed) with reshaped human peripheral blood mononuclear cells (PBMC), lymph nodes, fetal liver/thymus or other tissues, can be infected with lentiviral vector or HIV and used as models pathogenesis of HIV. Similarly, the simian immunodeficiency virus (SIV)/monkey model can be used as well as the feline immunodeficiency virus (FΓV)/cat model.

The pharmaceutical composition may contain other pharmaceuticals in combination with the vector of the invention when used for the therapeutic treatment of AIDS. These other pharmaceutical agents can be used in their conventional manner (ie, as antiviral agents for the treatment of HIV infection). Examples of anti-HIV agents include, without limitation, non-nucleoside reverse transcriptase inhibitors, protease inhibitors, entry or fusion inhibitors, and integrase inhibitors.

According to another embodiment, the present invention relates to an antibody-based pharmaceutical composition that contains an effective amount of an isolated anti-HIV antibody, or affinity matured version, which provides for a choice of prophylactic or therapeutic treatment in order to reduce infection with the HIV virus. The antibody pharmaceutical composition of the present invention may be formulated by any number of strategies known in the art (for example, see McGoff and Scher, 2000, Solution Formulation of Proteins/Peptides: In McNally, E.J., ed. Protein Formulation and Delivery. New York, NY: 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 administration to a patient contains an effective amount of the antibody in a formulation that retains biological activity while also facilitating maximum stability during storage within an acceptable temperature range. The pharmaceutical compositions may also contain, depending on the composition desired, pharmaceutically acceptable diluents, pharmaceutically acceptable carriers and/or pharmaceutically acceptable excipients, or any such excipients commonly used to formulate pharmaceutical compositions for administration to an animal or human. The diluent is chosen so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, physiological phosphate buffered saline, Ringer's solutions, dextrose solution, and Hank's solution. The amount of excipient that can be used in a 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 an individual in need. It may serve to dilute the antibody to a concentration that provides the desired beneficial palliative or curative results while minimizing any unwanted side effects that may result from too high a concentration. They may also have a preservative effect. Thus, for an antibody with high physiological activity, a larger amount of excipient is used. On the other hand, for any active ingredient(s) that exhibits lower physiological activity, a smaller amount of excipient is used.

The antibodies described above and antibody compositions or vaccine compositions containing at least one or a combination of the antibodies described herein can be administered for the prophylactic and therapeutic treatment of an HIV viral infection.

The present invention also relates to selected polypeptides containing the amino acid sequences of the light chains and heavy chains listed in table. A, B and in Fig. 10A-10C; heavy and light chain consensus sequences of SEQ ID NOs: 1 and 2 and insertion sequences of SEQ ID NOs: 3 and 4.

In other related embodiments, the invention relates to polypeptide variants that encode the amino acid sequences of anti-HIV antibodies listed in Table. A, B and in Fig. 10A-10C; heavy and light chain consensus sequences SEQ ID NOs: 1 and 2; and insertion sequences of SEQ ID NOs: 3 and 4. These polypeptide 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 more sequence identity compared to the polypeptide sequence of the present invention, as determined using the methods described herein, ( e.g. BLAST analysis using standard parameters). One of ordinary skill in the art will appreciate that these values can be adjusted accordingly in order to determine the appropriate identity of the proteins being encoded, considering amino acid similarity and the like.

The term polypeptide is used in its standard meaning, ie. amino acid sequence. The polypeptides are not limited to a particular product length. Peptides, oligopeptides and proteins including

- 12 042696 chens in the definition of a polypeptide, and such terms can be used interchangeably in this document, unless specifically indicated otherwise. The term also includes post-expression modifications of the polypeptide, such as glycosylation, acetylation, phosphorylation, and the like, as well as other modifications known in the art, both naturally occurring and non-naturally occurring. The polypeptide may be an entire protein or a subsequence thereof. Particular polypeptides of interest in the context of the present invention are amino acid subsequences containing CDRs, VH and V _L capable of binding an antigen or an HIV-infected cell.

A variant polypeptide, as the term is used herein, is a polypeptide that typically differs from a polypeptide specifically described herein by one or more substitutions, deletions, additions, and/or insertions. Such variants may be naturally occurring or may be created synthetically, 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 number of methods well known in the art. in this area.

For example, certain amino acids can be substituted for other amino acids in the structure of a protein without significant loss of its ability to bind other polypeptides (eg, antigens) or cells. Since the binding capacity and properties of a protein determine the biological functionality of that protein, certain amino acid sequence substitutions can be made in the sequence of the protein, and thus in the underlying DNA coding sequence, whereby a protein with similar properties is obtained. Thus, it is contemplated that various changes can be made to the peptide sequences of the disclosed compositions, or to the corresponding DNA sequences that encode said peptides, without significant loss of their biological utility or activity.

In many cases, the polypeptide variant contains one or more conservative substitutions. A conservative substitution is one in which an amino acid is replaced with another amino acid that has similar properties, such that one skilled in the art of peptide chemistry would expect the secondary structure and hydrophobic properties of the polypeptide to be substantially unchanged.

Amino acid substitutions are generally based on the relative similarity of the amino acid side chain substituents, eg, their hydrophobicity, hydrophilicity, charge, size, and the like. Exemplary substitutions that take into account the various characteristics above are well known to those skilled 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 a variant polynucleotide or polypeptide sequence that are identical to the non-variant sequence after sequence alignment and gaps, if necessary, to achieve the maximum percent homology. In specific embodiments, polynucleotide and polypeptide variants have a polynucleotide or polypeptide homology of at least about 70%, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least at least about 98% or at least about 99% with the polynucleotide or polypeptide described herein.

Such polypeptide variant sequences will have 70% or more (ie 80%, 85%, 90%, 95%, 97%, 98%, 99% or more) sequence identity with the sequences listed in the specification. In additional embodiments, the present invention relates to polypeptide fragments containing overlapping fragments of various lengths from the amino acid sequences described herein. For example, this invention provides peptide sequences that contain at least about 5, 10, 15, 20, 30, 40, 50, 75, 100, 150 or more overlapping peptides from one or more of the sequences described herein, and also all intermediate lengths in between.

The invention also includes nucleic acid sequences encoding part or all of the light and heavy chains of the disclosed patentable antibodies, as well as fragments thereof. Due to the redundancy of the genetic code, there are variants of these sequences that encode the same amino acid sequences.

The present invention also relates to selected nucleic acid sequences encoding polypeptides for heavy and light chains of antibodies against HIV, listed in table. A, B and in Fig. 10A-10C; heavy and light chain consensus sequences of SEQ ID NOs: 1 and 2 and insertion sequences of SEQ ID NOs: 3 and 4.

In other related embodiments, the present invention relates to polynucleotide variants that encode the peptide sequences of the heavy and light chains of the anti-HIV antibodies listed in Table 1. A, B and in Fig. 10A-10C; consensus sequences for heavy and

- 13 042696 light chains of SEQ ID NOs: 1 and 2 and insertion sequences of SEQ ID NOs: 3 and 4. 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 the polynucleotide sequence of this invention determined using the methods described herein (eg, BLAST analysis using standard parameters). One skilled in the art will appreciate that these values can be adjusted accordingly to determine the respective identity of the proteins encoded by the two nucleotide sequences, taking into account codon degeneracy, amino acid similarity, reading frame position, and the like.

The terms nucleic acid and polynucleotide are used interchangeably herein to refer to single or double stranded RNA, DNA, or mixed polymers. Polynucleotides may include genomic sequences, extra-genomic and plasmid sequences, and smaller engineered gene segments that express or can be adapted to express the polypeptides.

An isolated nucleic acid is a nucleic acid that is substantially separated from other genomic DNA sequences, as well as proteins or complexes, such as ribosomes and polymerases, that naturally accompany the native sequence. The term encompasses a nucleic acid sequence that has been removed from its natural environment and includes recombinant or cloned DNA isolates and chemically synthesized analogs or analogs biologically synthesized using heterologous systems. 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 that are later added to the isolated nucleic acid by human hands.

A variant polynucleotide, as the term is used herein, is a polynucleotide that typically differs from a polynucleotide specifically described herein by one or more substitutions, deletions, additions, and/or insertions. Such variants may occur naturally or be synthetically generated, for example, by modifying one or more polynucleotide sequences of the invention and evaluating one or more biological activities of the encoded polypeptide as described herein and/or using any number of methods well known in the art. this area.

It is possible to make modifications to the structure of the polynucleotides of the invention described and still obtain a functional molecule that encodes a polypeptide variant or derivative with the desired characteristics. When it is desired to change the amino acid sequence of a polypeptide in order to create an equivalent, or even improved, variant or portion of the polypeptide of the invention, one skilled in the art will typically change one or more codons of the DNA coding sequence.

Typically, variant polynucleotides contain one or more substitutions, additions, deletions, and/or insertions such that the immunogenic binding properties of the polypeptide encoded by the polynucleotide variant are not substantially impaired relative to the polypeptide encoded by the polynucleotide sequence specifically set forth herein.

In additional embodiments, the present invention relates to polynucleotide fragments that contain overlapping fragments of various lengths from a sequence identical or complementary to one or more of the sequences described herein. For example, provided herein are polynucleotides that contain at least about 10, 15, 20, 30, 40, 50, 75, 10°, 150, 200, 300, 400, 500, or 1000 or more contiguous nucleotides from one or more sequences. , described in this document, as well as all intermediate lengths in between, and cover any length between the specified 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 include all integers 200-500; 500-1000.

In another embodiment of the invention, polynucleotide compositions are provided that are capable of hybridizing under moderate to high stringency conditions with the polynucleotide sequence provided herein, or a fragment thereof, or a complementary sequence thereof. Hybridization techniques are well known in the art of molecular biology. By way of illustration, suitable moderate stringent conditions for testing hybridization of a polynucleotide of this invention with other polynucleotides include a pre-wash in a solution of 5x SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridization at 50-60°C, 5x SSC, overnight; followed by washing twice at 65°C. for 20 minutes using each of 2x, 0.5x and 0.2x SSC containing 0.1% SDS. One skilled in the art will appreciate that the stringency of hybridization can be easily manipulated, for example, by changing the salt content of

- 14 042696 solution for hybridization and/or temperature at which hybridization is carried out. For example, in another embodiment, suitable very stringent hybridization conditions include those described above, except that the hybridization temperature is raised to, for example, 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., binds specifically or preferentially to the same HIV epitope or strain) as the polypeptide encoded by the native polynucleotide. In some embodiments, the disclosed polynucleotides, variants, fragments, and hybridizable polynucleotide 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 of a polypeptide sequence as specifically set forth. in this document.

The polynucleotides of the 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, etc., so that they overall length can vary considerably. Nucleic acid fragments of almost any length are used. For example, exemplary polynucleotide segments with overall lengths of about 10,000, about 5,000, about 3,000, about 2,000, about 1,000, about 500, about 200, about 100, about 50 bp in length, and the like. (including all intermediate lengths) are included in many implementations of the present invention.

In some embodiments, the polynucleotide sequences provided herein are used as probes or primers for nucleic acid hybridization, for example, as PCR primers. The ability of such nucleic acid probes to specifically hybridize to a sequence of interest allows them to detect the presence of complementary sequences in a given sample. However, the invention described covers other uses, such as the use of sequence information to generate primers of a mutant species, or primers for use in the production of other genetic constructs. As such, nucleic acid segments of the invention that contain a sequence region of a contiguous sequence of at least about 15 nucleotides in length that has the same sequence as, or is complementary to, the 15 nucleotides in length contiguous sequence described herein are particularly useful. Longer contiguous identical or complementary sequences, e.g., about 20, 30, 40, 50, 100, 200, 500, 1000 (including intermediate lengths), including full-length sequences, and all lengths in between, are also used in some embodiments.

Polynucleotide molecules having sequence regions consisting of contiguous nucleotide fragments of 10-14, 15-20, 30, 50 or even 100-200 nucleotides or so (also including intermediate lengths) identical or complementary to the polynucleotide sequence described herein, in particular are considered as hybridization probes for use in eg Southern and Northern blotting and/or primers for use in eg PCR. The overall fragment size, as well as the size of the complementary fragment(s), ultimately depends on the intended use or application of the particular nucleic acid segment. Smaller fragments are generally used in hybridization embodiments where the length of the contiguous complementary region may vary, for example, between about 15 and about 100 nucleotides, but larger contiguous complementary fragments can be used, according to the lengths of the complementary sequences one wishes to detect.

The use of a hybridization probe of approximately 15-25 nucleotides in length allows the formation of a double molecule that is both stable and selective. Molecules having contiguous complementary sequences in fragments greater than 12 bases in length may be used, albeit in order to increase the stability and selectivity of the fusion and thereby improve the quality and degree of specificity of the resulting fusion molecules. Nucleic acid molecules can be used that have complementary gene fragments of 15-25 contiguous nucleotides or even longer where desired.

Hybridization probes are selected from any portion of any of the sequences described herein. All that is required is to review the sequences set forth herein, or any contiguous portion of sequences from approximately 1525 nucleotides in length up to and including the full length sequence that one wishes to use as a probe or primer. The choice of probe and primer sequences is determined by various factors. For example, one may wish to use primers towards the ends of the overall sequence.

- 15 042696

In addition, the scope of the invention includes vectors, such as expression vectors, which contain the nucleic acid sequence of the invention. Cells transformed with such vectors are also included within the scope of the invention.

The present invention also relates to vectors and host cells containing the nucleic acid of the invention, as well as to recombinant methods for producing a polypeptide of the invention. The vectors of the invention include those capable of replication in any type of cell or organism, including, for example, plasmids, phage, cosmids, and minichromosomes. In some embodiments, vectors containing a polynucleotide of the invention are vectors suitable for propagating or replicating a polynucleotide, or vectors suitable for expressing a polypeptide of the invention. Such vectors are known in the art and are commercially available.

The vector includes shuttle and expression vectors. Typically, the plasmid construct also contains an origin of replication (eg, ColE1 origin of replication) and a selectable marker (eg, ampicillin or tetracycline resistance) for plasmid replication and selection in bacteria, respectively. An expression vector refers to a vector that contains the necessary control sequences or regulatory elements for the expression of antibodies containing an 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 eukaryotic, multicellular species (e.g., as opposed to a unicellular yeast cell), such as, but not limited to, a mammalian cell or a human cell. The cell may be present as a single entity or may be part of a larger collection of cells. Such a larger collection of cells may include, for example, a cell culture (either mixed or pure), a tissue (e.g., endothelial, epithelial, mucosal, or other tissue), an organ (e.g., lung, liver, muscle, and other organs), an organ system ( eg, circulatory system, respiratory system, gastrointestinal tract, urinary system, nervous system, integumentary system, or other organ system) or an organism (eg, bird, mammal, or the like).

The polynucleotides of the invention can be synthesized, in whole or in portions, which are then combined, and inserted into a vector using standard molecular and cell biology techniques, including, for example, subcloning the polynucleotide into a linearized vector using appropriate restriction sites and restriction enzymes. The polynucleotides of the invention are amplified by polymerase chain reaction using oligonucleotide primers complementary to each polynucleotide strand. These primers also contain restriction enzyme cleavage sites to facilitate subcloning into the vector. Components of replicable vectors 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 selection genes.

In order to express a polypeptide of the invention, the nucleotide sequences encoding the polypeptide, or functional equivalents, can be inserted into a suitable expression vector, i. a vector that contains the necessary elements for transcription and translation of the inserted coding sequence. Methods well known to those skilled in the art can be used to construct expression vectors containing sequences that encode a polypeptide of interest and suitable transcriptional and translational control elements. These methods include in vitro recombinant DNA methods, synthetic methods, and in vivo genetic recombination. Such methods 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 & Sons, New York. N.Y.

The present invention also relates to kits that can be used in the implementation of diagnostic and prognostic analyzes using the antibodies, polypeptides and nucleic acids of the present invention. The kits of the present invention contain a suitable container that contains the anti-HIV antibody, polypeptide or nucleic acid of the invention in labeled or unlabeled form. In addition, when the antibody, polypeptide, or nucleic acid is supplied in a labeled form suitable for indirect binding assay, the kit further contains reagents for performing a suitable indirect assay. For example, the kit may contain one or more suitable containers containing enzyme substrates or derivatizers, depending on the properties of the label. Control samples and/or instructions may also be included. The present invention also relates to kits for detecting the presence of an anti-HIV antibody or an anti-HIV antibody nucleotide sequence of the present invention in a biological sample by PCR or mass spectrometry.

A label, as used herein, refers to a detectable compound or composition that is conjugated directly or indirectly to an antibody such that

- 16 042696 to create a labeled antibody. The label can also be conjugated to a polypeptide and/or nucleic acid sequence described herein. The label may itself be detectable (eg, radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze a detectable chemical change in the substrate compound or composition. The antibodies and polypeptides of the invention may also be modified to contain an epitope tag or tag, 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, fluorophores, chemiluminophores, chromogens, enzyme substrates or cofactors, enzyme inhibitors, prosthetic group complexes, free radicals, particles, dyes, and the like.

According to another embodiment, the present invention relates to diagnostic methods.

Diagnostic methods generally include contacting a biological sample obtained from a patient, such as blood, serum, saliva, urine, sputum, a cell scraping sample, or a tissue biopsy, for example, with an anti-HIV antibody and determining whether the antibody binds, preferably with a sample compared to a control sample or a predetermined threshold value, thereby indicating the presence of the HIV virus.

According to another embodiment, the present invention relates to methods for detecting the presence of the anti-HIV antibodies of the present invention in a biological sample from a patient. Detection methods generally include obtaining a biological sample from a patient, such as, for example, blood, serum, saliva, urine, sputum, a cell scraping sample, or tissue biopsy, and isolating anti-HIV antibodies or fragments thereof, or nucleic acids that encode anti-HIV antibodies, and analyzing the presence of anti-HIV antibodies in the biological sample. The present invention also relates to methods for detecting the nucleotide sequence of anti-HIV antibodies in a cell. The nucleotide sequence of an HIV antibody can also be detected using the primers described herein. The presence of an anti-HIV antibody in a biological sample from a patient can be determined using known recombinant methods and/or using a mass spectrometer.

In another embodiment, the present invention relates to a method for detecting an anti-HIV antibody containing a heavy chain that contains a highly conserved consensus sequence and a light chain that contains a highly conserved consensus sequence in a biological sample, which comprises obtaining an immunoglobulin-containing biological sample from a mammal, isolating the antibody against HIV from said sample and identifying highly conserved heavy chain and light chain consensus sequences. The biological sample may be blood, serum, saliva, urine, sputum, a cell scraping sample, or a tissue biopsy. Amino acid sequences can be determined by methods known in the art, including, for example, PCR and mass spectrometry.

The term evaluation includes any form of measurement and includes determining whether an element is present or not. The terms definition, measurement, evaluation, evaluation and analysis are used interchangeably and include quantitative and qualitative determination. The assessment can be relative or absolute. Presence assessment involves determining the amount of something present and/or determining whether it is present or absent. As used herein, the terms definition, measurement, evaluation, and analysis are used interchangeably and include both quantitative and qualitative determination.

II. A way to reduce virus replication.

In addition, methods are provided to reduce an increase in HIV virus titer, viral replication, viral proliferation, or the amount of HIV viral protein in an individual. In another aspect, the method includes administering to an individual an effective amount of an anti-HIV antibody to reduce an increase in HIV titer, viral replication, or the amount of HIV protein of one or more strains or isolates of HIV in the individual.

In another embodiment, the present invention relates to a method of reducing viral replication or the spread of HIV infection to additional host cells or tissues, which comprises contacting a mammalian cell with an antibody, or portion thereof, that binds to an antigenic epitope on gp120.

III. Method of treatment.

According to another embodiment, the present invention relates to a method of treating a mammal infected with a viral infection, such as HIV, which comprises administering to said mammal a pharmaceutical composition that contains the anti-HIV antibodies described herein. In one embodiment, a method for treating a mammal infected with HIV comprises administering to said mammal a pharmaceutical composition that contains an antibody of the present invention, or a fragment thereof. Compositions of the invention may contain more than one antibody having the disclosed characteristics (eg, a plurality or pool of antibodies). It may also include other neutralizing anti-HIV antibodies as known in the art, for example, but not limited to,

VRC01, PG9 and b12.

Passive immunization has been 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), each of which is incorporated herein by reference). Passive immunization using human monoclonal antibodies provides an immediate treatment strategy for acute HIV prevention and treatment.

Individuals at risk of HIV-related diseases or disorders include patients who have been in contact with an infected person or who have been exposed to HIV in some other way. Administration of the prophylactic may occur prior to the onset of symptoms characteristic of an HIV-related disease or disorder, such that the disease or disorder prevents or alternatively delays its progression.

For the treatment of human and non-human patients in vivo, the patient is administered or provided with a pharmaceutical formulation containing an anti-HIV antibody of the invention. When used for in vivo therapy, the antibodies of the invention are administered to a patient in therapeutically effective amounts (ie, amounts that eliminate or reduce the patient's viral load). Antibodies are administered to a human patient according to known methods, such as intravenous administration, for example, as a bolus or by continuous infusion over a period of time, via intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intraarticular, intrasynovial, intrathecal, oral, topical, or inhalation route. Antibodies can be administered parenterally, when possible, at the site of target cells or intravenously. In some embodiments, the antibody is administered via intravenous or subcutaneous administration. The therapeutic compositions of the invention may be administered systemically, parenterally, or locally to a patient or individual. The above parameters for assessing successful treatment and improvement in disease are easily measured by standard procedures familiar to the physician.

For parenteral administration, antibodies can be formulated in a standard injectable dosage form (solution, suspension, emulsion) together with a pharmaceutically acceptable, parenteral vehicle. Examples of such excipients include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous excipients include, but are not limited to, fatty oils and ethyl oleate. Liposomes can be used as carriers. The filler may contain minor amounts of additives such as substances that increase isotonicity and chemical stability, such as buffers and preservatives. Antibodies can be formulated in such vehicles at concentrations of from about 1 to 10 mg/ml.

The dose and dosing regimen depend on various factors which are readily determined by the physician, such as the nature of the infection, eg its therapeutic index, the patient, and the patient's history. In general, a therapeutically effective amount of the antibody is administered to the patient. In some embodiments, the amount of antibody administered is in the range of about 0.1 to about 50 mg/kg of the patient's body weight. Depending on the type and severity of the infection, from about 0.1 to about 50 mg/kg body weight (e.g., about 0.1-15 mg/kg/dose) of antibody is the initial dose considered for administration to a patient, for example, or via one or several separate introductions or by continuous infusion. Monitoring the dynamics of therapy is easy to carry out using standard methods and analyzes, as well as based on criteria known to the doctor or other specialists in this field. The above parameters for assessing successful treatment and improvement of the disease can be easily measured using standard procedures familiar to the physician.

Other therapeutic regimens can be combined with the administration of an anti-HIV antibody of the present invention. Combined administration includes co-administration, use of separate formulations or a single pharmaceutical formulation, and sequential administration in any order, where preferably there is a period of time during which both (or all) active agents simultaneously exhibit their biological activities. Such combination therapy may lead to a synergistic therapeutic effect. The above parameters for assessing successful treatment and improvement of the disease can be easily measured by standard procedures familiar to the physician.

The terms treatment or alleviation are used interchangeably and refer to both therapeutic treatment and prophylactic or preventive measures; where the goal is to prevent or slow down (alleviate) the target condition or disorder. Those in need of treatment include those who already have the disorder, as well as those who are prone to having the disorder, or those whose disorder is to be prevented. An individual 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 an observable and/or measurable decrease in

- 18 042696 or the absence of one or more of the following: a decrease in the number of infected cells or the absence of infected cells; a decrease in the percentage of infected cells among all cells and / or a certain degree of relief of one or more symptoms associated with a particular infection; reduced morbidity and mortality; and improved quality of life. The above parameters for assessing the success of treatment and improvement of the disease are easily measured by standard procedures familiar to the physician.

The term therapeutically effective amount refers to the amount of an antibody or drug effective to treat a disease or disorder in an individual or mammal.

Administration in combination with one or more additional therapeutic agents includes simultaneous (competitive) and sequential administration in any order.

Carriers, as used herein, include pharmaceutically acceptable carriers, excipients, or stabilizers that are not toxic to the cell or mammal when exposed to the doses and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffer solution. Examples of physiologically acceptable carriers include, but are 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) polypeptides; 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 non-ionic surfactants such as, but not limited to, TWEEN; polyethylene glycol (PEG) and PLURONICS.

Where ranges of values are provided, it is understood that the invention covers every intermediate value, up to a tenth of one of the lower limit, unless the context clearly dictates otherwise, between the upper and lower limits of that range and any other declared or intermediate value within that declared range. . The invention also includes the upper and lower limits of these ranges, which may be independently included in smaller ranges, provided that no specifically excluded limit is included in that declared range. Where a declared range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as they are commonly understood by a person skilled in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are described. All publications cited in this document are incorporated herein by reference in their entirety.

As used herein and in the appended claims, the singular forms include the plural unless the context clearly dictates otherwise.

The publications described herein are provided for disclosure only prior to the filing date of the present invention. Nothing herein is to be taken as an admission that the present invention cannot backdate such publication to an earlier invention. In addition, the publication dates provided may differ from the actual publication dates, which may require independent confirmation.

Each of the applications and patents cited in this text, as well as each document or literary source, patent or non-patent literature cited in each of the applications and patents (including during the maintenance of each patent granted; documents cited in the application), and each of the PCT and foreign applications or patents corresponding to and/or claiming priority for any of those applications and patents, and each of the documents cited or referred to in each of the documents cited in the application are expressly incorporated herein in as reference in its entirety. More generally, documents or literature are cited in this test, or a list of literature cited before the claims; or in the text itself; and each of those documents or literature sources (literature references cited herein), as well as each document or literary source cited in each of the literature sources cited herein (including any descriptions, manufacturer's instructions, etc.), are hereby expressly incorporated herein by reference.

The following non-limiting examples serve to further illustrate the present invention.

- 19 042696

Example 1

Materials, methods and equipment.

Samples.

Human samples were collected after informed consent was signed in accordance with protocols agreed upon by the Institutional Review Board (IRB) by all participating institutions. Patient 1 was selected from a non-progressor cohort administered at the Aaron Diamod AIDS Research Center, New York. Patients 3 and 8 were selected from a selective controller group administered at the Ragon Institute in Boston. Patients 1, 3 and 8 were selected based on their broad serum neutralizing activity against a standard panel of HIV isolates. Patient 12 was selected from Protocol G cohort of the International Aids Vaccine Initiative based on broad serum neutralizing activity.

Staining, sorting single cells and cloning antibodies.

Individual cells were stained and sorted with 2CC core and gp140 specific Ig+ memory B cells (J.F. Scheid et al., Nature, 458, 636 (Apr 2, 2009)). Briefly, CD19+ B cells were enriched from peripheral blood mononuclear cells using anti-human CD19 MACS magnetic beads (Miltenyi Biotec) and subsequently stained with anti-human CD20 and anti-human IgG antibodies (Becton Dickinson) and biotinylated 2CC-core (B Dey et al., PLoS Pathog 5, e1000445 (May, 2009)) or YU2-gp140 trimer (R. Diskin, P. M. Marcovecchio, P. J. Bjorkman, Nat. Struct. Mol. Biol. 17, 608 (May, 2010)) followed by detection using streptavidin-related phycoerythrin (PE, Beckton Dickinson). Individual cells were sorted on a FACSAria III cell sorter (Becton Dickinson), excluding cell duplicates, into 96-well PCR plates (Denville) containing 4 μl/well of ice-cold 0.5x phosphate-buffered saline (PBS) containing 10 mM DTT, 8 units RNAsin® (Promega), 0.4 units 5'-3' Prime RNAse Inhibitor™ (Eppendorf). The plates were sealed with Microseal® 'F' (BioRad) film, immediately frozen on dry ice prior to storage at -80°C.

cDNA synthesis and Ig amplification was performed (H. Wardemann et al., Science, 301, 1374 (Sep 5, 2003)) with the following modifications.

Instead of using the original primer sets, the first and second immunoglobulin-specific PCR was performed using the primers described in table. 1, in a semi-nested approach. The heavy and light chain PCR products were cloned into their respective expression vectors, and 100% identity of the cloned expression plasmids with the original PCR product was confirmed by sequencing prior to antibody expression in HEK 293 cells.

ELISA.

96-well high binding ELISA plates (Costar) were coated overnight with 100 ng/well of purified antigens (gp140, gp120, gp41, gp120 ^core and 2CC-core) (B. Dey et al., PLoS Pathog 5, el000445 ( May, 2009)) and mutant proteins (gp120 D368R, gp120 I420R) in PBS. After washing, the plates were blocked for 2 hours with 2% BSA, 1 μM EDTA, 0.05% Tween-PBS (blocking buffer) and then incubated for 2 hours with IgG antibodies diluted 4 μg/ml and several subsequent 1:4 dilutions in PBS . After washing, the plates were developed by incubation for 1 hour with horseradish peroxidase-conjugated goat anti-mouse IgG (Jackson ImmunoReseach) (0.8 μg/ml in blocking buffer) and by adding 100 μl of horseradish peroxidase chromogenic substrate (ABTS solution, Invitrogen). Optical densities were measured at 405 nm (OD405 _nm ) using an ELISA microplate reader (Molecular Devices). Subtracted background values obtained by incubation only with PBS in coated wells. IgG antibodies were tested for polyreactivity (H. Mouquet et al., Nature, 467, 591 (Sep 30, 2010)), and were considered polyreactive when they recognized at least two structurally different antigens out of four tested; ssDNA, dsDNA, insulin and LPS. Reactivity thresholds were determined using control antibodies mGO53 (negative), eiJB40 (weak positive) and ED38 (strong positive).

Neutralization assay: A neutralization screen was performed (D.C. Montefiori, Curr Protoc. Immunol. Chapter 12, Unit 12 11 (Jan, 2005)). Briefly, neutralization was detected as a decrease in luciferase reporter gene expression after one round of infection in Tzm-bl cells. In order to rule out non-specific antiviral activity in antibody samples, MuLV (mouse leukemia virus) was used as a negative control.

Clone-specific identification of bone marrow plasma cells.

Bone marrow plasma cells were stained with anti-human CD138 and anti-CD19 antibodies (Becton Dickinson) after Ficoll purification of mononuclear cells from bone marrow aspirates using Ficoll-Paque (GE Healthcare). Mass sorting of CD138+ CD19+ human plasma cells was carried out on a FACSAriaIII cell sorter (Becton Dickinson) and RNA isolation was performed per 100,000 cells using Trizol LS reagent (Invitrogen) according to the manufacturer's instructions. The RNA was reverse transcribed using Superscript III reverse transcriptase (Invitrogen) according to the manufacturer's instructions. The cDNA was then subjected to

- 20 042696 immunoglobulin-specific PCR with the following modifications: 1 μl of cDNA was amplified in 2 rounds of nested clone-specific immunoglobulin heavy chain PCR using the first round of the forward leader primer and reverse constant region primer presented in table. 1 followed by clone-specific forward and reverse primers designed based on sequencing results from single cell analysis. PCR products were gel purified and cloned into TOPO TA vectors (Invitrogen) according to the manufacturer's instructions. Colonies were screened by PCR with clone-specific primers and sequenced.

Surface plasmon resonance.

All experiments were performed using Biacore T100 (Biacore, Inc) in HBS-EP+ running buffer (Biacore, Inc) at 25°C as previously described (Mouquet 2010). YU-2 gp140 and 2CC-core proteins at a concentration of 12.5 μg/ml were immobilized on CM5 chips (Biacore, Inc.) by amine binding at pH 4.5, leading to an immobilization level of 100 RU. For kinetic measurements on gp140- and 2CC-core-derivatized chips, IgG was injected through flow cells at 700 nM and 4 serial 1:2 dilutions in HBS-EP+ running buffer (Biacore, Inc.) at flow rates of 40 μl/min for 3 min. association and 5 min dissociation. The sensor surface was regenerated between each experiment using a 30 second injection of 10 mM glycine-HCl pH 2.5 at a flow rate of 50 μl/min. Dissociation rate (kd (c ^-1 )), association rate (ka (M ^-1 s ^-1 ) and binding constants (KD (M) or KA (M ^-1 ) were calculated after subtracting background signals (binding to control flow cells and HBS-EP+ running buffer signal) using Biacore T100 Evaluation software using kinetic analysis and a 1:1 binding model. Interaction Analysis, University of Utah).

CD4i site induction.

293T was transfected with gp160 ^BAL . ²⁶ Ac or gp160 ^YU.2 Ac in a pMX-IRES-GFP construct (Pietzsch et al. 2010) using Fugene™6 (Roche) at a plasmid:Fugene ratio of 1:2. After 48 hours, 293T cells were washed in PBS and detached using trypsin-free cell dissociation buffer (Gibco) and resuspended at 10 ⁷ cells/ml in FACS buffer (1 x PBS, 2% FBS, 2 mM EDTA). sCD4 (Progenies Pharmaceuticals, Inc.) and mAb was added to gp160-expressing 293T cells in a 1:4 dilution series starting at a final concentration of 40 μg/mL. mGO is a negative control antibody that does not bind to gp160Δc (H. Mouquet et al., Nature, 467, 591 (Sep 30, 2010)). After a 15 min incubation on ice, cells were divided and stained for 25 min on ice with Alexa647 labeled CD4-induced mAbs (3-67; (JF Scheid et al., Nature, 458, 636 (Apr 2, 2009)) or Alexa647 labeled control mAbs (i.e. PG16; LM Walker et al., Science, 326, 285 (Oct 9, 2009)) or 2G12 for gp160 ^YU.2 and 2G12 for gp160 ^BAL.26 ). Antibody labeling was performed using an Alexa Fluor® 647 Microscale Protein Labeling Kit (Invitrogen). Cells were analyzed on a LSRFortessa cell analyzer (BD Bioscience).

Crystallization.

IgG 3BNC60 was expressed by transient expression in HEK293-6E cells and a Fab fragment was obtained by papain digestion (R. Diskin, P.M. Marcovecchio, PJ Bjorkman, Nat. Struct. Mol. Biol. 17, 608 (May, 2010). Crystallization screening was performed at 20°C by vapor diffusion in nanoliter sessile drops using a Mosquito™ crystallization robot (TTP LabTech) on MRC crystallization plates (Jena Bioscience) Fab 3BNC60 at 9.5mg/mL was combined with reservoir solution at a ratio of 1: 1 to create 400 nl drops Initial crystallization hits were obtained using a PEGRx HT™ (Hampton Research) crystallization sieve and further optimized by hand Crystals suitable for data collection grew after several weeks in 11.7% polyethylene glycol 20000.0 .1 M sodium acetate pH 5.0, 100 mM potassium/sodium tartrate, 20 mM lithium sulfate, 10 mM N-cyclohexyl-2-aminoethanesulfonic acid (CHES) pH 9.5 in monoclinic symmetry group P2 ₁ with two Fab in the asymmetric element. The crystals were soaked in a 15% glycerol reservoir solution for 2 hours before being immersed in a 30% glycerol reservoir solution and flash-cooled in liquid nitrogen. Diffraction data were collected in a 12-2 Stanford Synchrotron Radiation Lightsource (SSRL) channel at 100 K using a Pilatus 6M detector. Data were indexed, integrated and scaled using XDS W. Kabsch, Acta Crystallogr D Biol Crystallogr 66, 125 (Feb, 2010) (Table 8). Molecular displacement was performed using a Phaser with the V _H and _CH 1 domains from the antitumor antibody CTM01 (PDB code 1AD9) and with the VL and CL domains of the anti-gp120 b13 antibody (PDB code 3IDX) as search models. The construction and refinement of models to a resolution of 2.65 A was performed iteratively using Phenix P. Emsley, B. Lohkamp, WG Scott, K. Cowtan, Acta Crystallogr. D Biol. Crystallogr. 66, 486 (Apr, 2010) and Coot (P. Emsley, B. Lohkamp, W.G. Scott, K. Cowtan, Acta Crystallogr. D Biol. Crystallogr. 66, 486 (Apr, 2010)). The structure was refined using the maximum likelihood objective function and non-crystallographic symmetry constraints. The final model (R _work =20.7%; R _free =25.7%) contains 6478 atoms

- 21 042696 proteins, 146 water molecules and 28 sugar atoms (Table 8). 91.9%, 7.6%, and 0.5% of the residues were in the favorable, acceptable, and unacceptable areas of the Ramachandran map, respectively. Structural analysis and visualization were performed using PyMol (The PyMOL Molecular Graphics System, version 1.3, Schrodinger, LLC). The structure of 3BNC60 consists of residues 3-205 for the light chain (including the first N-acetylglucosamine in the N-linked carbohydrate attached to Asn72) and 2-217 for the heavy chain. The order of residues at terminal residues and residues 133-140 in the CH1 domain was broken.

Mass spectrometry.

IgG was purified from serum using ProteinG Sepharose (GE Healthcare) according to the manufacturer's instructions. IgG was then digested with immobilized papain (Pierce) and digested mixtures of Fab-Fc fragments were incubated with saturating amounts of biotinylated 2CC-core protein. Streptavidin bound to Dynabeads (Invitrogen) was added after a 15 min incubation at room temperature and subjected to 10 rounds of PBS (Gibco) washes. Bound Fab fragments were eluted with lithium dodecyl sulfate buffer (Invitrogen) at 95° C. and sample purity was confirmed by SDS polyacrylamide gel electrophoresis followed by silver staining or Coomassie staining prior to analysis by mass spectrometry.

Isolated Fab fragments were reduced with dithiothreitol, alkylated with iodoacetamide, resolved by 1D gel electrophoresis on a 4-12% NuPAGE Novex Bis-Tris gel (Invitrogen) and stained with Coomassie blue (Thermo Fisher). Fab fragments were excised from the gel and digested with 200 ng trypsin (Promega). The resulting peptides were isolated using a reverse phase resin (PORS 20 R2, Applied Biosystem) and eluted using an aliquot of 40% acetonitrile in 0.5% acetic acid and a second aliquot of 80% acetonitrile in 0.5% acetic acid. Acetonitrile was removed using a SpeedVac (Thermo Fisher Scientific) and aliquots of the remaining solution were loaded under pressure onto a self-sealing PicoFrit® column (New Objective, Woburn, MA) with an inline emitter head (360 µm O.D., 50 µm I.D., 10 µm head) packed with 6 see reverse phase C18 material (ReproSil-Pur C18-AQ, 3 µm beads from Dr. Maisch GmbH) and mated to an Agilent 1200 Series HPLC system (Agilent) or an LTQ Orbitrap™ XL mass spectrometer or LTQ Orbitrap Velos mass spectrometer ™ (Thermo Fisher Scientific) using a homemade microelectrospray source. The peptides were eluted into the mass spectrometer with the following gradient: 0 to 5% B over 5 min, 40% B over 125 min, 60% B over 150 min, 100% B over 165 min (A = 0.1 M acetic acid, B = 70% acetonitrile in 0.1 M acetic acid, flow rate 90 Nl/min). Both instruments were operated in data dependent mode and both mass spectrometers were set to a target value of 5e5 ions and a resolution of 60,000 (at 400 m/z). For LTQ Orbitrap™ XL analysis, a full scan was followed by 8 MS/MS scans for the 8 most common ions from that full scan. Peptides (only charge states >1) were isolated with a window of 2 Da, target window of 1e4 of ions dissociated through CAD (normalized collision energy = 35, Q activation = 0.25, activation time = 30 ms) and mass analyzed in LTQ. For LTQ Orbitrap™ Velos analysis, a full scan was followed by 10 MS/MS scans at 7500 resolution for the 10 most common ions from the immediately preceding full scan. Peptides (only charge state >2) were isolated with a 3 Da window, target window of 2e5 ions dissociated through HCD (normalized collision energy = 40, activation time = 0.100 ms) and mass analyzed in Orbitrap. For any instrument, ions selected for MS/MS were put on the exclusion list for 30 s. The obtained MS/MS spectra were searched in the Human IPI database and the patient specific IgG database using Xtandem!, the peptides were automatically compared with tryptic peptides in the Human IPI database and the patient specific database. Peptide matches corresponding to specific patient IgGs were manually verified.

Multiple sequence alignment.

All multiple sequence alignments were performed using CLUSTALW2 with default parameters (weight matrix: GONNET for proteins and UIB for DNA, gap opening = 10, gap extension = 0.1). Shaded alignments were created using the TeXshade package.

alignment consensus.

Consensus sequences for multiple sequences were generated based on identity and similarity between residues (>=70%). Amino acids were grouped by similarity as follows: FYW, ILVM, RK, DE, GA, ST, and NQ.

Phylogenetic germ trees.

The relationship between the sequences was created using the method of closest linking. A bootstrap consensus tree, which was obtained from 1000 replicas, was taken to represent kinship. Branches were collapsed corresponding to splits reproduced in less than 50% of the bootstrap replicas. The percentage of replicated trees where related sequences clustered together in the bootstrap test (1000 replicas) is shown after the branches. Trees are drawn to scale, with branch lengths in the same units as the evolutionary distances used to build the phylogenetic tree. Evolutionary distances were calculated using the difference number method and are given in units of the number of amino acid differences per sequence. All indeterminate positions were removed for each pair of sequences. Evolutionary analysis was performed in MEGA5.

R/S ratio calculation.

The DNA sequences were superimposed on protein alignments to calculate the substitution/substitution ratio. All gap positions were removed from the analysis. The R/S ratio was analyzed using Perl scripts.

Example 2

Isolation of antibodies against HIV.

In order to determine if cloning of anti-HIV antibodies is limited due to somatic mutations, a new set of primers was designed to avoid this potential problem (Table 1). The new primer set was tested by sorting B cells that bind to the HIV core protein gp120, which lacks V1-3 loops and contains a pair of stabilizing disulfide bonds (2CC-core). Unlike the resurfaced primer used to clone VRC01, the 2CC-core primer also allows antibody capture at the CD4-induced co-receptor binding site (CD4i).

In a head-to-head comparison, the new primer set increased the yield of IgH strands compared to the initial primer set (FIG. 4A). Antibodies generated using the new primer set contained more mutations (mean 35.6 vs 19.8, p=<0.0001 and maximum 85 vs 50 for IgH) and contained clones that were not found using the original primer set ( Fig. 4A). To determine if the new primers rescued VRC01-like antibodies from cells that were sorted using YU2 gp140, frozen cDNA samples were tested from an individual who had already been carefully screened using the original primer set without producing any VRC01-associated clones. Three antibodies were found in 80 wells corresponding to the VRC01 variants defined by the IgH and IgL sequences (FIGS. 5A and 5B). It was concluded that VRC01-like antibodies captured the gp140 trimer, and that primers that were specifically designed to clone highly mutated antibodies captured a relatively large fraction of anti-HIV antibodies from memory B cells of patients with high titers of broad-spectrum neutralizing antibodies.

Four unrelated HIV-infected individuals, including 2 European, 1 Spanish and 1 African donors, showing high titers of broad-spectrum neutralizing antibodies were tested using 2CC-core primer, including 2 individuals whose pre-cloned antibodies could not explain their serological activity (Table .2 and figures 6A and 6B). 576 antibodies representing 201 different unique and heterogeneous clones were generated from an initial population of 1.5x105 IgG ⁺ B memory cells (Table 3).

Example 3

Anti-HIV antibody binding specificity.

The size of the antibody clones captured by the 2CC-core primed varied widely in the range of 2-76 heterogeneous members (Table 3). To determine if antibodies captured by the 2CC-core bound to the HIV spike, an ELISA was performed using YU2 gp120 on representative members of each propagated clone. All tested antibodies bound to gp120 (Table 3).

The antibody binding site on the HIV spike was mapped using mutant proteins that affect either CD4bs (gp120 (D368R)) or the CD4-induced co-receptor binding site (CD4i, gp120 (I420R)). As reported by X. Wu et al., Science, 329, 856 (Aug 13, 2010), VRC01 was classified as an anti-CD4bs antibody because it is sensitive to the D368R mutation, but because of the proximity of the CD4i site, it also shows some sensitivity to the I420R mutation . NIH45-46 which is a variant of VRC01 and antibodies 3BNC60, 8ANC131 and 12A12 showing ELISA patterns that are similar to VRC01 (these clonal members were selected based on neutralizing activity, Table 3). Other clones, including 1B2530 and 8ANC195, are equally susceptible to both mutations and cannot be accurately classified based on ELISA alone.

To determine if the antibodies are polyreactive, ELISA was performed on purified ssDNA, dsDNA, insulin and LPS. 63% of the tested antibodies against 2CC-core were polyreactive. It has been concluded that most of the antibodies captured by 2CC primer recognize either CD4bs or the CD4i site on gp120 and many are also polyreactive.

Example 4

Somatic hypermutation.

Somatic hypermutation is required for the development of high antigen binding affinity and in some cases it contributes to the polyreactivity of anti-HIV antibodies. In order to test if this is the case, specific antibodies against 2CC-core with a high level of mutations, four representative antibodies were reversed into the appropriate germline. Reversal led to complete loss of antigen binding in ELISA for all four clones tested and loss of polyreactivity.

Example 5

Neutralization of HIV.

HIV neutralizing activity was measured in standardized in vitro assays using an initial panel of eight viruses containing three tier 1 clades A, B and C and five tier 2 clades B Env pseudovirus variants (M.S. Seaman et al., J. Virol. 84, 1439 (Feb, 2010). The neutralizing activity of the antibodies was compared with VRC01 and purified serum IgG from donors (Figure 1A, Table 4 and Figure 6). Antibodies showing high levels of neutralizing activity were further tested in a panel of 15 additional tiers of 2 clades A, B, C, D, G, AG, and AE Env pseudovirus variants (Figure 1B, Table 5), including five viruses that are resistant to VRC01 (Fig. 1B and Table 5).

90% of all antibodies tested showed some neutralizing activity and six clones contained antibody variants that showed high levels of potency and breadth (FIGS. 1A-C and Tables 4 and 5). These clones were also the most abundant among those captured by 2CC primer in each of the four patients studied (Table 3). The most impressive of the new antibodies, 3BNC117, a 76 member clone, showed a mean IC ₈₀ on the combined layer 14 2 virus group of 0.5 µg/mL compared to 1.8 µg/mL for VRC01 (Fig. 1C, Table .4 and 5).

Only 4 of the 20 viruses tested were more sensitive to VRC01 than 3BNC117, while 14 were more sensitive to 3BNC117, including DU172.17, which is completely resistant to VRC01 but susceptible to 3BNC117 (Fig. 1B and C). NIH45-46, a new variant of VRC01, is more effective than VRC01 on 15 out of 20 viruses tested, but still less effective than 3BNC117 (Figures 1B and C and Tables 4 and 5).

There was significant variation in the width and efficiency of neutralization among members of the five clones of the most effective neutralizing antibodies. For example, 3BNC156, a variant of 3BNC117, neutralized only two viruses in the initial panel, and at significantly higher concentrations than 3BNC117 (Fig. 1A and Table 4) and 3BNC55, the other variant, was between two others showing activity against six viruses at average IC ₅₀ 4 μg/ml (Fig. 1 and table. 4). Finally, the most active antibodies had a high level of hypermutations. The average number of mutations for the top 10 antibodies was 72 for V _H and 45 for V _L and this is related to their breadth and potency (Tables 4 and 5). Reversal of mutated residues to the germline led to a complete loss of neutralizing activity for all antibodies tested.

Example 6

Identification of diagnostic peptides.

The above cloning strategy captured antibodies produced by antigen-binding memory B cells, but these cells did not produce circulating antibodies and instead they originated from plasma cells in the bone marrow. However, a similar antigen cannot be used as a primer for uptake by plasma cells because they do not express surface Ig A. (Radbruch et al., Nat. Rev. Immunol. 6, 741 (Oct, 2006)). In addition, the relationship between plasma cells in the bone marrow and circulating memory B cells is not well defined. In order to determine whether antibodies cloned from memory B cells were also found in the bone marrow plasma cell compartment, CD138-expressing plasma cells were purified from paired bone marrow samples from 2 of the 4 subjects studied and PCR was used to specifically amplify the IgV _H genes for more effective antibodies cloned from memory B cells in these individuals. The following are clone-specific primers for RU01:

CTGCAACCGGTGTACATTCTCAAGTGCAACTGGTGC (straight) (SEQ ID

NO:584), CTGCAACCGGTGTACATTCTCAGGTCCATTTGTCACAG (straight), (SEQ

ID NO:585) TGCGAAGTCGACGCTGACGAGACAGTGACCTGC (reverse) (SEQ ID

NO:586), TGCGAAGTCGACGCTGAAGAGACAATAATTTG (reverse) (SEQ ID

NO:587), TGCGAAGTCGACGCTGACGAGACAATAACT (reverse) (SEQ ID

NO:588) and for RU10: CTGCAACCGGTGTACATTTTCAGGGGCACTTGGTG (direct) (SEQ ID NO:589), TGCGAAGTCGACGCTGAGGTGACGATGACCGTG (reverse) (SEQ ID NO: 590) .

Members of selected clones and a large number of additional variants were easily identified in both patients.

In order to confirm that these antibodies can also be found in serum, IgG purified from the sera of the same two and one additional individual was adsorbed onto a 2CC-core primer and mass spectrometry of the eluted IgG was performed (FIGS. 1D, 7 and 10A-10C ). In all cases, diagnostic peptides for highly active antibody variants were found (Fig. 7, 10A-10C). We concluded that effective broad-spectrum anti-HIV antibodies cloned from memory B cells were also found in the bone marrow plasma cell compartment and in circulating IgG of patients with

- 24 042696 high titers of broad-spectrum neutralizing antibodies in serum.

Example 7

Binding characteristics of an anti-HIV antibody.

To determine if antibody affinity for gp120 is related to neutralizing activity, the binding of highly active antibodies, selected clonal relatives, and reverted germline progenitors were compared using surface plasmon resonance (SPR) (FIGS. 2A and 2B, 8 and Table 6). ).

The best neutralizing antibodies showed affinities (KA) in the range ξ10 ⁷ -10 ¹² (M ^-1 ) on the YU2 gp140 and ^107-10 ¹¹ (M ^-1 ) trimers on the 2CC core (FIGS. 2A and 2B and Table 6) . Consistent with their reported neutralizing potency and width, 3BNC66, 3BNC156, and 3BNC55 showed lower affinities for gp140 YU2 trimers than 3BNC117, but surprisingly, affinities for the 2CC core did not correlate with neutralizing activity (FIGS. 1, 8, Table 4 and 6). SPR binding was not detected for any of the germline reverted antibodies tested (FIG. 2B, Table 6). It has been concluded that anti-HIV antibodies captured by the YU2 2CC-core tend to show higher affinity for the corresponding gp140 trimer than for the 2CC-core.

When VRC01 binds to the HIV spike, it creates a large conformational change that mimics CD4 binding and exposes the CD4i site. In contrast, b12 and most other known anti-CD4bs antibodies do not cause this.

To determine if this is a common feature of highly active antibodies, HIV-BAL.26AC or -YU2 gp160Δc was expressed on the surface of HEK 293T cells and CD4i antibody binding was measured in the presence or absence of CD4 or anti-CD4bs antibodies (Fig. 2C). With one exception, all highly active antibodies tested were similar to CD4 and VRC01 in that they facilitated binding of the anti-CD4i antibody to HIV-BAL.26 or YU2 gp160Δc or both (FIG. 2C).

The only highly active antibody that did not share this common feature, 8ANC195, was not a conventional anti-CD4bs antibody in that it was equally sensitive to D368R and I420R mutations (Table 3). In addition, it differed from other highly active antibodies in its neutralization pattern: it does not neutralize any of the tier 1 viruses and shows effective activity against H086.8, a clade B virus that is resistant to all other antibodies tested, including 3BNC117, VRC01 and b12 ( Fig. 1A and B and tables 4 and 5).

Example 8

Sequence identity of anti-HIV antibodies.

In order to determine whether highly active anti-CD4bs antibodies share sequence features, the top 10 antibodies: Two variants, each of five independently generated antibody clones from five different patients, were aligned (FIG. 3). Comparison of IgV _H regions revealed a highly conserved consensus sequence spanning 68 IgVH residues (FIG. 3A). The IgVH consensus contained six contact residues of VRC01-gp120, including VRC01-Arg 71, which mimics the key interaction between Arg59 _CD4 and Asp368 _gp120 (Fig. 3A). In addition, the six-contact consensus is completely conserved in both closely related IgVH germline genes (VH1-2 and VH1-46) that give rise to all antibodies in this class (FIGS. 3A and 3B).

The codons encoding the consensus residues had a high level of somatic mutations in the ten selected antibodies, however, the amino acid sequences were conserved (FIG. 9). The ratio of substitutions to silent mutations in the consensus residues was in the range of 0.71.7, while it was 3.5-22 in the non-consensus residues, indicating that the conservatism of the consensus is the result of rigorous selection (Table 7). Unlike the heavy chain, the VRC01 light chain only makes 8 of the total 32 contacts with gp120. Consistent with its more limited role, comparison of the light chain sequences of the same antibodies revealed a less extensive consensus covering 53 IgV _L residues, including three VRC01-gp120 contact residues (FIG. 3B). Finally, like heavy chains, light chains arose from a limited set of germline genes: 2 were derived from IgK1D-33, 2 from IgK3-11, and one from IgL1-47 (Figure 3B and Table 3). The 8ANC195 antibody, which differed from the others in several important respects, did not fit the consensus completely and did not originate from related heavy or light chains (FIGS. 3A and 3B). It has been concluded that there is significant sequence convergence among highly active anti-CD4bs agonist antibodies (HAAD).

Example 9

Crystal structure of Fab 3BNC60.

In order to determine whether the antibody structure of different patients is also conserved, the crystal structure of Fab 3BNC60 was resolved with a resolution of 2.65 A and compared with VRC01. The structure revealed four domains V _H , _CH 1 , V _L and C _L of the classic Fab and complementarity determining regions (CDRs) in V _H and V _L that form the antigen binding site. The two Fabs in the 3BNC60 asymmetrical element were almost identical; however, the conformation of residues 74-78 in the loop,

- 25 042696 connecting strands D and E, varied slightly due to the different chemical environment formed by the contacts of the crystal lattice.

The superposition of VH domains from 3BNC60 and VRC01 in the VRC01-gp120 co-crystal structure (T. Zhou et al., Science, 329, 811 (Aug 13, 2010)) gave a standard deviation (rmsd) of 1.3 A (calculated for 111 Ca atoms ) with the main differences in CDR2 residues 58-65 (numbering 3BNC60). The structure overlay showed the conservatism of the contact area recognizing gp120. For example, Arg72 _3BNC60 adopted a similar conformation to that of Arg71 _VRC01 , which mimics an important salt bridge normally formed between Arg59 _CD4 and Asp368 _gp120 . In addition, Trp47 _3BNC60 adopted a similar conformation as Trp47 _VRC01 , a residue that contacts gp120 and is involved in a complex network of aromatic and aliphatic residue interactions that stabilize the CDRH3 and CDRL3 conformations. Gln65 _3BNC60 , which corresponds to Gln64 _VRC01 , is in a segment of residues (residues 58-65) that differs in structure from VRC01. The conformation of this region of 3BNC60, which is involved in lattice contact in the crystals, is likely to change upon gp120 binding, as it will collide with the CD4-binding loop on gp120.

The superposition of the V _L domains of 3BNC60 and VRC01 gave an rmsd of 0.9 A (calculated for 95 Ca atoms) and showed that some of the residues in contact with gp120 are structurally conserved; Tyr91 _3BNC60 and Glu91a _3BNC60 adopted similar conformations as Tyr91 _VRC01 and Glu96 _VRC01 that contacted the D loop of gp120 via polar interactions. Overall, these structural comparisons show that 3BNC60 binds gp120 with the same architecture as observed for VRC01 binding.

Example 10

The consensus sequence of an anti-HIV antibody.

In the experiments above, a class of anti-CD4bs agonist antibodies, HAAD, were defined that share IgV _H and IgV _L consensus sequences, including eight contact residues between VRC01 and the HIV spike (FIGS. 3A and 3B). In five different donors, selected for their high levels of anti-HIV serology, these antibodies originated from only two closely related IgVH and three IgVL germline genes that correspond to the HAAD consensus: VH1-2 and VH1-46 differ in only 7 amino acids, none of which is not part of the consensus (Fig. 3A). Despite extensive somatic hypermutation, the consensus residues were preserved in their germline form.

The only exception to the consensus, 8ANC195, differed from the others in a variety of ways, suggesting that it may have a unique way of recognizing the antigen: the absence of Arg in the heavy chain, which mimics the required contact site for Arg59 _CD4 and Asp368 _gp120 ; unique neutralizing pattern; and failure to promote anti-CD4i antibody binding. This antibody is one of two different highly active antibodies occurring in the same patient, providing further support for the idea that serologic neutralizing activity is combinatorial.

- 26 042696

Table A

SEQ ID NO: Antibody Heavy chain amino acid sequence 5 8A253HC QGQLVQSGGGLKKPGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTAYNFQDRLSLRRDRSTGTVFM ELRGLRPDDTAVYYCARDGLGEVAPDYRYGIDVWGQGS TV IVTAASTKG 6 8A275HC QGLLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 7 8ABM11 FQGHLVQSGGGVKKPGTSVTLSCLASEYTFTEFTIHWIRQ APGQGPLWLGLIKRSGRLMTSYRFQDRLSLRRDRSTGTVF MELRSLRTDDTAVYYCARDGLGELAPAYHYGIDAWGQGTT VIVTSASTS 8 8 ABM 12 QGHLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSAST 9 8ABM13 QGHLVQSGGGVKKLGTSVTISCLASEDTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTS 10 8ABM14 GHLVQSGGGXKKPGTSVTISCLASEYTFTEFTIHRIRQAP GQGPLWLGLIKGSGRLMTSYGFQDRLSLRRDRSTGTVEME LRSLRTDDTAVYYCARDGLGELAPAYHYGIDVWGQGTTVI VTSASTS

- 27 042696

eleven 8ABM2 0 GVHFQGHLVQSGGGVKKPGSSVTISCLASEYTFTEFTIHW IRQAPGQGPLWLGLIKRSGRLMTSYRFQDRLSLRRDRSTG TVFMELRGLRIDDTAVYYCARDGLGEVAPAYLYGIDVWGQ GTTVIVTSASTS 12 8ABM2 4 FQGQLVQSGGGVKKPGSSVTISCLASEYTFTEFTIHWIRQ APGQGPLWLGLIKRSGRLMTSYGFQDRLSVRRDRSTGTVF MELRSLRTDDTAVYYCARDGLGELAPAYHYGIDVWGQGTT VIVTSASTS 13 8ABM26 QGQLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTS 14 8ABM27 QGHLVQSGXEVKKPGSSVKVSCKASGGTFSXYAIGWVRQA PGQGLEWMGGIIPILGT TNYAQRFQGGVTITADE S TNTAY MDVS S LRS DD TAVYYCAKAPYRPRGS GNYYYAMDVWGQGT TVIVSSASTS 15 8ANC105HC QGHLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 16 8ANC116HC QGQLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVSSASTKG 17 8ANC127HC QGHLVQSGGGVKKLGTSVTISCLVSEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 18 8ANC131HC QGQLVQSGGGLKKPGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTAYNFQDRLRLRRDRSTGTVFM ELRGLRPDDTAVYYCARDGLGEVAPDYRYGIDVWGQGS TV IVTAASTKG 19 8ANC134HC QGQLVQSGGGVKKPGTSVTISCLASEYTFNEFVIHWIRQA PGQGPVWLGLIKRSGRLMTSYKFQDRLSLRRDRSTGTVFM ELRGLRLDDTAVYYCARDGLGEVAPAYLYGIDAWGQGS TV

- 28 042696

IVTSASTKG 20 8ANC13HC QGQLVQSGGGVKKPGASVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTAYKFQDRLSLRRDRSTGTVFM ELRGLRPEDTAVYYCARDGLGEVAPDYRYGIDVWGQGSTV IVSAASTKG 21 8ANC171HC QGHLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 22 8ANC18 GVHFQGHLVQSGGGVKKPGSSVTISCLASEYTFTEFTIHW IRQAPGQGPLWLGLIKRSGRLMTSYRFQDRLSLRRDRSTG TVFMELRGLRIDDTAVYYCARDGLGEVAPAYLYGIDVWGQ GSTVIVTSASTS 23 8ANC182HC QGQLVQSGGGVKKPGTSVTISCLASEYTFTEFTIHWIRQA PGQGPLWLGLIKRSGRLMTAYRFQDRLSLRRDRSTGTVFM ELRNLRMDDTAVYYCARDGLGELAPAYQYGIDVWGQGTTV IVSSASTKG 24 8ANC192HC QGHLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 25 8ANC22HC QGHLVQSGGGVKKLGTSVTISCLASEDTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 26 8ANC26HC QGQLVQSGGGVKKPGTSVTISCLASEYTFNEFVIHWIRQA PGQGPVWLGLIKRSGRLMTSYKFQDRLSLRRDRSTGTVFM ELRGLRLDDTAVYYCARDGLGEVAPAYLYGIDAWGQGSKV IVTPASTKG 27 8ANC2HC QGQLVQSGGGVKKLGTSVTIPCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 28 8ANC30HC QGQLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM

- 29 042696

ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 29 8ANC37HC QGHLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG thirty 8ANC40HC QGHLVQSGGGVKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVTSASTKG 31 8ANC41HC QGQLVQSGGGVKKTGTSVTISCLASEYTFTEFTIHWIRQA PGQGPLWLGLIKRSGRLMTANRFQDRLSLRRDRSTGTVFM ELRSLRIDDTAVYYCARDGLGELAPAYHYGIDVWGQGTTI IVTSASTKG 32 8ANC45HC QGQLVQSGGGVKKTGTSVTISCLASEYTFTEFTIHWIRQA PGQGPLWLGLIKRSGRLMTANRFQDRLSLRRDRSTGTVFM ELRSLRIDDTAVYYCARDGLGELAPAYHYGIDVWGQGTTI IVTSASTKG 33 8ANC50HC QGQLVQSGGGVKKPGTSVTISCLASEYTFTEFTIHWIRQA PGQGPLWLGLIKRSGRLMTAYRFQDRLSLRRDRSTGTVFM ELRNLRMDDTAVYYCARDGLGELAPAYQYGIDVWGQGTTV IVSSASTKG 34 8ANC53HC QGQLVQSGGGGKKLGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYQFQDRLSLRRDRSTGTVFM ELRGLRVDDTAVYYCARDGLGEVAPAYLYGIDAWGQGTTV IVSSASTKG 35 8ANC88HC QGQLVQSGGGVKKPGTSVTISCLASEYTFNEFVIHWIRQA PGQGPLWLGLIKRSGRLMTSYKFQDRLNLRRDRSTGTVFM ELRGLRPDDTAVYYCARDGLGEVAPDYRYGIDVWGQGS TV IVTAASTKG 36 8ANC103HC QVQLQQWGSGLLKPSETLSLTCAVYGGSFRSYYWNWIRQS PGKGLEWIGEVSHSGSTNYNPALKSRVTISVDTSKNQFSL KVKSVTAADTALYYCSRGRGKRCSGAYCFAGYFDSWGQGG LVWSSASTKG 37 8ANC106HC E VQLVE S GGGWE PGE S LRL S CAAS GET FRS YDMFWVRQA

- 30 042696

TGKSLEWVSAIGIAGDTYYSGSVKGRFTISRENARTSLYL QLSGLRVEDSAVYFCVRGSPPRIAATEYNYYYGLDVWGQG TTVSVFSASTKG 38 8ANC107HC VVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRYVVNWVRQA PGQGLEWMGGMIPIFGIAKYAQKFQDRVTMTADESKNTVY LDFSSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 39 8ANC108HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRYVVNWVRQA PGQGLEWMGGIIPIFGIAKYAQKFQDRVTMTADEPKNTVY LDFNSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 40 8ANC109HC EVQLVESGGGLVKPGGSLRLSCAASGFSFSEHYMSWIRLA PGKGLEWLSYISSSTRTTYSADSVRGRFTISRDTAKQLLF LHMSSLRAEDTAVYYCVRLYGGINGWFDQWGQGTLVSVSS ASTKG 41 8ANC10HC QVQLVQSGAEVKKPGSSVKVSCKTSGGSFSNYAFSWVRQA PGEGLEWMGRIIPIFGTAKYTQKLQGRVTITADKFTSTVY MELSSLRSEDTAIYYCASLHQGPIGYTPWHPPPRAPLGQS VCG 42 8ANC111HC QVQLVE S GAEVKKPGASVKVS CKAS GYTFTSHDINWVRQA TGQGLEWMGWMNPNSGDTGYAHKFQGRVTMTRNTPISTAY MELSSLRSEDTAVYYCARGRATSRNTPWAHYYDSSGYYGA GDYWGQGTLVTVS SAS TKG 43 8ANC112HC QVQLVE S GGGWQPGRS LRL FCAAS G FAFNT YGMHWVRQA PGKGLEWVAVTWHDGSQKYYADSVKGRFTISRDNSKNTLY LQMNSLRAEDTAIYYCASDQGGFDDSSGYFAPGGMDVWGR GTTVIVSSAPTKG 44 8ANC113HC QVQLVE S GAE LRKPGE S LEIS CKAS GYS FS S HWIGWARQM PGKGLEWMGIIYPGDSNTIYS PS FQGQVTISADKSINTAY LQWSSLKASDTAMYFCASNYHDYFYWGQGTLVTVSSASTK G 45 8ANC114HC EVQLVESGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PGQGLEWMGG11PIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG

- 31 042696

46 8ANC115HC QVQLVQSGAE VRKPGSSLKVSCKSSGGTFSRYWNWVRQA PGQGLEWMGG11PIEGIAKYAQKFQDRVTMTADE PKNTVY LDFNSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 47 8ANC117HC EVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PGQGLEWMGG11PIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG 48 8ANC11HC QVFVQLVQSGGGLVQPGGSVRLSCTASGFLFSTYSMNWVR QAPGKGLEWVSSISTTSNYIYYADSVKGRFTISRSNGQGS LYLQLNSLRVEDTAVYYCARDTKVGAPRQDCYAMDLWGQR DHGHRLLSFHQGPIGLPPGALLQ 49 8ANC121HC QVQLLESGPGLVTPSGTLSLACAVSGASISSSHWWTWVRQ SPGKGLEWIGEIDRRGTTNYNPSLRSRVTILLDNSKNQFS LSLRSVTAADTAVYYCTKVYAGLFNERTYGMDVWGHGTTV LVSSASTKG 50 8ANC126HC QVQLVE S GAE VRKPGS S LKVS CKS S GGT FS 51 8ANC130HC QVQLLQSGAEVKKPGASVKVSCKVSGYTLTELSINWVRQA PGKGLEWMGGFDPEDDEAIYEPKFQGRLTMTTEDTTDTAY MELSSLRSEDTAVYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 52 8ANC132HC QVQLVQSGTEVQKPGASVKVSCKTSGYTFSKYYIHWVRQA PGQGLEWVGRINTDSGGTDYAEKFQGRVTMTRDTSITTVY LEMRGLTSDDTAAFYCARGPPHAGGWTIYYYGLDVWGQGT SVIVSSASTKG 53 8ANC133HC VELSSLRSEDTAMYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 54 8ANC136HC E VQLVE S GGGWQPGRS LRL S CAAS GFTFSHHGIHWVRQA PGEGLEWVAVISEDGTNIHYEDSVRGRFTISRDNSKNTVD LQMNSLRAEDTAVYYCASLISMRDGDAFDLWGQGTRVTVS

- 32 042696

SASTKG 55 8ANC137HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRYWNWVRQA PGQGLEWMGG11PIEGIAKYAQKFQDRVTMTADE S KNTVY LDFSSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 56 8ANC139HC QVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQA PGKGLEWVS SISSSSSYIYYADSVKGRFTISRDNAKNS LY LQMNSLRAEDTAVYYCAREGSYYYGMDVWGQGTTVTVSSA STKG 57 8ANC140HC EVQLVQS GGGLVQPGRS LRLS CAAS GET FDDYAMHWVRQA PGKGLEWVSGISWNSGTIGYADSVRGRFTISRDDAKSSLY LQMNSLRTEDTALYYCAKDGWVGSGSTLRGSDYWGQGTL VTVSSASTKG 58 8ANC142HC QIHLVQSGTDVKKPGSSVTVSCKAYGVNTFGLYAVNWVRQ APGQSLEYIGQIWRWKSSASHHFRGRVLISAVDLTGSSPP ISSLEIKNLTSDDTAVYFCTTTSTYDQWSGLHHDGVMAFS SRGQGTLISVSAASTKGPSVFPLAPSSKSTYGLAHVL 59 8ANC143HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRYWNWVRQA PGQGLEWMGG11PIEGIAKYAQKFQDRVTMTADE PKNTVY LDFNSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 60 8ANC144HC ASTKG 61 8ANC145HC QVQLLE S GGGLVQRGGS LRL S C TAS G FVFNNYWMTWVRQA PGNGLEWVANIDQDGSEKHYLDSVKGRFTISRDNAKNSLY LQMNSLRAEDTAIYYCARVRFKVTAWHRFDSWGQGDLVTV SSTSTKG 62 8ANC146HC LVQLLQSGAEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKGLEWMGGFDPEDDEAIYEPKFQGRLTMTEDTSTDTAY MELSSLRSEDTAVYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 63 8ANC147HC QVQLVESGGGLGQPGGSLRLSCAASGFTFRNYAMSWVRQA AGKGLEWVSGVSGGGDTTYYGDSVKGRFTISRDNSKNTLY

- 33 042696

LQMNNLRAEDTAVYYCAKDKGVWGSSDFDYWGQGTLVTVS SASTKG 64 8ANC148HC QVHLVQSGAEVKKPGASVRVSCKASGYTFTTYGISWVRQA PGQGLEWMGWISAHSGDTNYAQKLQARVTMTTDTSTNTAY MELRSLTSDDTAVYYCARDRPRHYYDRGGYYSPFDYWGQG TLVTVSSASTKG 65 8ANC149HC SASTKG 66 8ANC14HC QVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKGLEWMGGFDSEDGEAFYKQNFQGRVTMTEDTSTDTAY MELRRLRSEDTAVYYCATADRFKVAQDEGLFVIFDYWGQG TLVTVSSASTKG 67 8ANC150HC QVQLLQSGGEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKGLEWMGGFDPEDDEAIYEPKFQGRLTMTEDTSTDTAY MELSSLRSEDTAVYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 68 8ANC151HC EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQA PGKGLEWVSYISGSSYTIYYADSVRGRFTISRDNAKNSLY LQMNSLRDEDTAVYFCARATPPNPLNLYNYDSSGSSFDYW GQGTLVTVSSASTKG 69 8ANC153HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRYWNWVRQA PGQGLEWMGGMIPIFGIAKYAQKFQDRVTMTADESKNTVY LDFSSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 70 8ANC154HC QVQLVE S GAE VRKPGS S LKVS CKS S GGT FS 71 8ANC155HC QVQLVQS GAEIKKPGE S LKISCKAS GYT FNDYWIGWVRQM PGKGLEWMGIFYPDDSDSNYSPSFQGRVTISADKSITTAY LQWSTLKASDSAMYFCARLLGDSGAFDIWGQGTMVIVSSA STKG 72 8ANC156HC EVQLVESGAEVRKPGSSLKVSCKSSGGTFSRFWNWVRQA

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PGQGLEWMGGMIPIFGIAKYAQKFQDRVTMTADESKNTVY LDFSSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 73 8ANC157HC QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PGQGLEWMGG11PIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG 74 8ANC158HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRFWNWVRQA PGQGLEWMGGMIPIFGIAKYAQKFQDRVTMTADESKNTVY LDFSSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 75 8ANC160HC QVQLVQS GGGWQPGRS LRL S CAAS GFTFSHHGIHWVRQA PGEGLEWVAVISEDGTNIHYEDSVRGRFTISRDNSKNTVD LQMNSLRAEDTAVYYCASLISMRDGDAFDLWGQGTRVTVS SASTKG 76 8ANC161HC EVQLVQSGGGLVKPGGSLRLSCAASGFTFKNAWMSWVRQA PGKGLEWVGHIKSKTDGGTIDYAAPVKGRFTISRDDSKNT LYLQMNSLKIEDTAVYYCTTDIGSGRGWDFHYYDSNDWGQ GTLVTVSSASTKG 77 8ANC162HC EVQLVQSGGGWQPGRSRLSCWSGFTFSSFTFHWVRQA PGKGLEWVAGMSFHATYIYYADSVKGRFTISRDDSQDTLY LEMDSLRSEDTAIYYCARDPGIHDYGDYAPGAFDYWGQGS PVTVSSASTKG 78 8ANC163HC LVQLVQSGAEVKKPGASVKVSCKVSGHTLSELSINWVRHV PGKGLEWMGGLDPEDGEAIHEPKFQGRLTMTTEDTSDTAY STLSVWAPVAAAMYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 79 8ANC164HC EVQLVESGAEVKKPGSSVKVSCKASGGTFSSYSISWVRQA PGQGLEWMGGIIPIFATTHYGQKFQGRIKITADKSTSTAY MELSRLRSEDTAVYYCARDREFYFYGMDVWGQGTTVTVSS ASTKG 80 8ANC165HC QVQLQQWGAGLLKPSETLSLTCAVYAGSFSGYYWTWIRQP PGKGLEWIGEVNHGGSTNYNPSLKSRVTLSVDTSKNQFSL KLTSVTAADTAVYYCARVSRYDFWSGNYGSYGLDVWGQGT TVTVSSASTKG

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81 8ANC166HC WQLVQSGAEVRKPGSSLKVSCKSSGGTFSRFWNWVRQA 82 8ANC168HC LVQLVQSGAEVKKPGASVKVSCKVSGYSLTELSIHWVRQA PGKGLEWMGGFDSEDGEAIYKQNFQGRVTMTEDTSTDTAY MELSRLRSEDTAVYYCATADPFKVAQDEGLFVIFDYWGQG TTGHRLLSLHQGPHRLYSLGTLLSRAPIVQTHMA 83 8ANC169HC QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PGQGLEWMGG11PIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG 84 8ANC16HC QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PIEGLEWMGG TIPIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG 85 8ANC173HC QVQLVQWGAGLLKPLETLSLTCAVYGGSFNGYFWSWIRQT PGKGLEWIGEINHGGSANFNPSLKSRVTMSVDTSKNQFSL KLASVTAADTAIYYCARGRITMVRGDPQRGGVRMDVWGQG TSVTVSSASTKG 86 8ANC174HC QVQLMQSGAEVKRPGASVKVSCKAFRHSLNNLGISWIRRA PGRGLEWLGWINVYEGNTKYGRRFQGRVTMTTDRSTNTVS MELRSLTSDDTAVYYCARDNHFWSGSSRYYYFGMDVWGQG TTVIVSSASTKG 87 8ANC175HC QVQLVQS GGGLVQPGE S LRL S C TAS G FT FS S YNMNWVRQA PGKGLEWISYISDKSKNKYYADSVRGRFTISRDNAQNSLF LQMSSLRDEDTAVYYCTREGPQRSFYFDYWGQGIQVTVSS ASTKG 88 8ANC176HC QVQLQESGPGLVKPSETLSLTCTVSGGSISNHYWSWIRQP PGKGLEWIGYIYHSGNINYKSSLKSRATISIDTSNNQFSL KLS SVIAADTAVYYCARNFGPGS PNYGMDVWGQGT TVTVS SASTKG 89 8ANC177HC WQLVQSGPGLVKPSQTLSLTCTVSGGSISSGDFYWSWIR QPPGKGLEWIGYIYYSGSTYYNPSLKSRLTISVDTSKNQF SLRLSSVTAADTAVYYCARDLNSRIVGALDAFDIWGQGTM

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VTVSSASTKG 90 8ANC178HC QVQLVESGGALVQPGGSLRLSCAASGFSFSSYAMSWVRQA PGKGLEWVSAISRSGGS TYYADSVKGRFTISIDNSNNTLY LQMNSLRVEDTAVYYCAKREAFYYGAGGYGMDVWGQGTTV TVSSASTKG 91 8ANC179HC EVQLVE S GGGLVKPGGS LRL S CEAS G FT FTNAWMNWVRQA PGKGLEWVGRIKSQTHGGTTRYAAPVKGRFTISRDDSKHT LYLQMDRLTTEDTAVYYCTGTITGSTFYYYGMDVWGQGTT VTVSPASTKG 92 8ANC17HC EVQLVESGGGLLQPGGSLRLSCAASGFSFNDFEMNWVRQA PGKGLEWVSYISNDGTMIHYADSVKGRFTISRDNAKKSLF LQMNSLRAEDTAVYYCARLAEVPPAIRGSYYYGMDVWGQG TTVTVASASTKG 93 8ANC180HC QVQLQESGPGLLRPLETLSLTCSVSGGSIRGYFWSWVRQP AGRGLEWIGRIYSSGTTRFNPSLKSRVRLSIDTAKSEVSL NITSVTAADSASYFCAGTSPVHGGLDLWGLGLRVTVSSAS TKG 94 8ANC181HC HLVQSGTEVKKPGSSVTVSCKAYGVNTFGLYAVNWVRQAP GQSLEYIGQIWRWKSSASHHFRGRVLISAVDLTGSSPPIS SLEIKNLTSDDTAVYFCTTTSTYDQWSGLHHDGVMAFSSW GQGTLISVSAASTKG 95 8ANC184HC EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKGLEWMGGFDPEDDEAIYEPKFQGRLTMTTEDTTDTAY MELSSLRSEDTAVYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 96 8ANC185HC QVQLVESGGGLVQPGGSLRLSCAASGFTFSTHWMHWVRQA PGKGLVWVSRIHSDGRSTSYADSVKGRFTISRDNAKNTLY LQMNSLRAEDTAVYYCARGAAVFGWIIGGMDLWGQGTTV TVSSASTKG 97 8ANC186HC E VQ L VE S G G G WQ P G G S L RL S CAAS G FM FKN YAMH W VRQ P 98 8ANC187HC EVQLVESGGGLIQRGGSLRLSCVASGFPVSDNHMSWVRQA PGKGLEWVSIIYSDGGTYYADSVKGRFTISRDNSKNTVYL

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QMNSLRATDTAVYYCARDPGFHYGLDVWGQGTTVTVSSAS TKG 99 8ANC188HC WQLVESGGGLVQPGGSLRLSCAASGFAFRSYWMSWVRQA PGRGLEWVANIKQDGSEKYYADSVKGRFTISRDNTKNSLY LQMNSLRAEDTAVFYCASRGDRYGPIDYWGQGTLVTVSSA STKG 100 8ANC191HC WQLVESGTEVKKPGSSVKVSCKASGGTFSGSDISWVRQA PGQGLEWMGGIIPMFDIENHAEKFRGRLTITAVKSTGAAY MELSSLRSEDAAVYYCARSSGNYDFAYDIWGQGTRVIVSS AS TKG 101 8ANC193HC EVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PGQGLEWMGG TIPIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG 102 8ANC194HC EVQLVQSGGGLVQPGGSLRLSCAASGLTFRNFAMSWVRQA PGKGLEWVS SISGSGGS TYYADSVKGRFTISRDNSKNTLY LQMNSLRGEDTAVYFCAKGVGYDILTGLGDAFDIWGQGTV VAVSSASTKG 103 8ANC195HC QIHLVQSGTEVKKPGSSVTVSCKAYGVNTFGLYAVNWVRQ APGQSLEYIGQIWRWKSSASHHFRGRVLISAVDLTGSSPP ISSLEIKNLTSDDTAVYFCTTTSTYDKWSGLHHDGVMAFS SWGQGTLISVSAASTKG 104 8ANC196HC WQLVQSGTEVKKPGSSVKVSCKASGGTFSGSDISWVRQA PGQGLEWMGGIIPMFDIEDHAQKFRGRLTITADKSTGAAY MELSSLRSEDAAVYYCARSSGNYDFAFDIWGQGTRLIVSS AS TKG 105 8ANC20HC QVQLGESGGGLVEPGGSLRLSCAASGFLFSDYQMSWIRLA PGKGLEWIS FISGFGSVYYADSVEGRFTISRDNARNSLYL QMNNLRAEDTAVYYCARAYGTGNWRGLYYYYYGMDVWGHG TTVTVSSASTKG 106 8ANC21HC QLQLVESGGGWQPGRSLRLSCAASGFTFSTYTMHWVRQA PGKGLEWVAVISYDGTNKYYADSVKGRFTISRDNSKNTLY LQMNSLRGEDTAVYYCARSPSYYFDYWGQGTLVTVSAAST KG 107 8ANC24HC QVQLVQSGAEVKMPGASVKVSCKVSGYSLTELSIHWVRQA

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PGKRLEWMGGFDPEDDERIYAQKFQDRVTMTEDTSTDTAY MDLNSLRSEDTAVYYCTTGGLYCSSISCIMDVWGQGTTVI VSSASTKG 108 8ANC25HC QVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKRLEWMGGFDPEDGERIYAQKFQGRVTMTEDTSTDTAY MELNSLRSDDTAVYYCATGGLYCSSISCIMDVWGQGTTVT VSSASTKG 109 8ANC27HC QVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKGLEWMGGFDSEDGEAIYKQNFQGRVTMTEDTSTDTAY MELSRLRSEDTAVYYCATADRFKVAQDEGLFVIFDYWGQG NPGHRLLSLHQGPIGLPPGTLPPKATSGHAARR 110 8ANC31HC QVQLVE S GGGWQPGRS LRL S CAAS G FT FS S YAMHWVRQA PGKGLEWVAVISYDGSNKYYADSVKGRFTISRDDSKS TVY LQINSLRAADTAVYFCAREGGLRFLEWLFWGQGTLVTVSS GESSASTKG 111 8ANC33HC EFQLVQSGGGLVKPGGSLRLSCTGTFFSFSSDDMNWVRQA PGKGLEWVSSMSDSGSHIYYADFVKGRFTISRDNAKKSLY LQMNSLRAEDTAVYYCAQSRPPQRLYGMDVWGQGTTVTVS SASTKG 112 8ANC34HC QVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKGLEWMGGFDPEDGEASFEPKFQGRLTMTEDTSTDTAY MELSSLRSEDTAVYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 113 8ANC36HC QVQLVESGGGWQPGKSLRLSCAASGFTFSTHAMHWVRQA PGKGLDWVAVISHDGDNQYYADAVKGRFTISRDDSRDTVF LQMNSLRTEDTGVYYCAADSSGSNWFDYWGQGILVTVSSA STKG 114 8ANC38HC EPMFQPGQSGGVVVQSGESLHLSCEASGFKFASQMMHWVR HVPGRGLEWVALISWDGS GKL FADSVRGRFTIHRWNDRNS LYLDVKNVRPEDAAIYYCTRNGFDVWGQGILVTVSSASTK G 115 8ANC39HC QVQLLQSGAEVKKPGASVKVSCKVSGYTLTELSIHWVRQA PGKGLEWMGGFDPEDDEAIYEPKFQGRLTMTEDTSTDTAY MELSSLRSEDTAVYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG

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116 8ANC3HC QVHLQESGPRLVRSSETLSLTCSVPGGSIVNPITNYYWSW IRQSPRKGLQWIGDIYYTGTSRNPSLDSRVSISMDVSRK QISLTLYSVTAADTAVHYCASQSLSWYRPSGYFESWGQGI LVTVSSASTKG 117 8ANC43HC QVQLVQSGAEVKKPGSSMKVSCKSSGGTFSNHAISWVRQA PGKGLEWMGGIIPMSGTTNYLQKFQGRVTITADEFATTAY MELSSLTSEDTAVYYCARARADSHTPIDAFDIWGPGTRVI VSSASTKG 118 8ANC46HC QVQLVQSGTEVKKPGSSVKVSCKASGGTFSDSDIAWVRQA PGQGLEWMGGIT PMFDMAKSAQKFRGRL11TADKS TGTAY MELSSLRYEDAAVYFCARSSGNFEFAFEIWGQGTKIIVSL ASTKG 119 8ANC48HC QVQLVQS GAEVKKPGASVKVS CKAS GYTFTSYDINWVRQA TGQGLEWMGWMNPNSGNTGYAQTFQGRVTMTRNTSISTAY MELSSLRSEDTAVYYCARDRWLPQYYYYGMDVWGQGTTVT VSSASTKG 120 8ANC49HC FVQLVESGGGLVQPGGSLRLSCAASGFNFNTYWMNWVRQA PGKGLEWVANMKEDGSEKYYVDSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARNPESRCIVGRNRGWCRYFDLWG RGSLVTVSPASTKG 121 8ANC51HC LVQLVE S GGGWQPGRS LRL S CAAS G FT FS 122 8ANC57HC PMFQPGQSGGVVVQSGESLHLSCEASGFKFASQMMHWVRH VPRGLEWVALISWDGSGKLFADSVRGRFTIHRWNDRNSL YLDVKNVRPEDAAIYYCTRNFDVWGQGILVTVSSASTKG 123 8ANC58HC VELSSLRSEDTAMYYCATADPFKVAQDEGLYVIFDYWGQG TLVTVSSASTKG 124 8ANC5HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRFWNWVRQA PGQGLEWMGGMIPIFGIAKYAQKFQDRVTMTADESKNTVY LDFSSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG

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125 8ANC60HC LVQLVESGGGWQPGKSLRLSCATSGFTFSTYGMHWVRQA PGKGLEWVAVIWYDGSYKYYADSVKGRFTISRDNSKNTLF LQMNSLRAEDTAMYYCGREMAVGGTKALDHWGQGTLVTVS SASTKG 126 8ANC63HC QVQLVQSGAEAKRPGDSVKVSCKASGYTFTEYYIHWVRQT PGQGFEWMGIITPGAGNTTYAQKFQGRITVTRDTSAATVY MELSNLTSEDTAVYFCSRGVSFWGQGTLVTVSSASTKG 127 8ANC65HC QVQMVASGGGLVKPGGSLRLSCEASGFTFSDYYMSWVRQA PGKGLEWISYITSGGNALYYADSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDLLHAHDFGRQGTLVTVSSAST KG 128 8ANC67HC Q VQ L VE S G G WQ P GR S LRLSCATSGFTSKNY G VH W VRQA 129 8ANC69HC QVQLVQS GAEVKKPGASVKVS CKAS GYT FT DYYIHWVRQA PGQGLEWMGWINPSTGGTNFVQKFLGRVTMTSDTSINTAY MELRRLKNDDAAIYYCATYSTRQFFHYYYVTDVWGQGTTV TVSSASTKG 130 8ANC6HC QVQLVQSGAEVKKPGSSVKVSCRASGGSFGNYAINWVRQA PMQGLEWMGGIIPIFGTTNYAQNFRGRVTINADTTFTNTVN MDLSSLRSEDTAVYYCGRSINAAVPGLEGVYYYYGMAVWG QGTTVTVSSASTKG 131 8ANC70HC QVQLHQWGAGLLKPSDTLSLTCGILGVSPPGSLTGYYWTW IRQPPGKGLEWIGEVYHSGSTNYNPSLASRVTISMGTTKT Q FS LRL T SVTAADSAVYYCAS GKVWGITARPRDAGL DVWG QGTTVTVTSASTKG 132 8ANC71HC EVQWESGGGLVQPGGSLRLSCVASGFTFSEYWMSWVRQA PGKGLEWVATIKRDGSEESYVDSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARVRDPNYNLHFDSWGQGTLVTVS SASTKG 133 8ANC72HC QVQLVE SGGGLIQPGGSLRLS CEAS G FAVGDINYMS WVRQ APGKGLEWVSVLYSGGSSQYADSVKGRFTISRDNSRNTLY LQMDNLRAEDTAVYYCARGLRVYFDLWGQGILVTVSSAST KG

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134 8ANC74HC QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PGQGLEWMGG11PIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG 135 8ANC75HC QVQLQESGPGLVKPSETLSLTCTVSGGSISSRSYYWGWIR QPPGKGLEWVGSIYYTGSTYYSPSLKSRVTISVDTSQNQF SLKLNSVTAADTAVYYCARQKGSGTSLLYWGQGTLVTVSS AS TKG 136 8ANC76HC QVQLVQSGSELKKPGASVKVSCKASGYTFTSYAINWVRQA PGQGLEWMGWINTNTGNPTYAQGFTGRFVFSLETSVSTAY LQINSLKAEDTAVYYCARDLLESRTYYNDIRDCWGQGTLV TVSSASTKG 137 8ANC78HC QVQLQESGSGLVKPSGTLSLTCAVSNGPISSGNWWSWVRQ TPEKGLEWIGEVYHSGSTNHNPSLKSRATILVDKSKNQFS LNLRSVTAADTAVYYCARVRGSWNFDYWGQGILVTVS SAS TKG 138 8ANC79HC QHQLVPCVAEVRKPGASVKVSCKVSGYTLTEISMHWVRQA PGKGLEWMGGFDREDGETIYAQKFQGRVTMTEDTSTDTAY MELSSLRSEDTAVYYCATTYLAWPDGFDGYSSSWYWFDP WGQGTLVTVSSASMQGPMLLSPTGTLLPRAPLVQTRPGP 139 8ANC7HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRYWNWVRQA PGQGLEWMGGIIPIFGIAKYAQKFQDRVTMTADESKNTVY LDFSSLRSDDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGTTVIVSSASTKG 140 8ANC80HC QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAFSWVRQA PGQGLEWMGG11PIFGTENYAQKFQGRVTITADKS T S TAY MELSSLRSEDTAVYYCARDRSSAIGYCSSISCYKGSFDIW GQGTMVTVSSASTKG 141 8ANC82HC QVHLEESGPGLVKTSQTLSLTCSVSSYSISRSGYFWTWIR QRPGKGLEWIGYIYFNGRTTYNPSLKSRITISRDTSHSQF SLTLNSLSAADTAVYYCGRCQDGLASRPIDFWGQGTLVTV SSASTKG 142 8ANC83HC QVQLVE S GGGWQPGKS LRLSCAISGFL FNNYGGQWVRQA PGKGLEWVAAISYDGNNRYYADSAKGRFLISRDTPKNILY LQIYSLRLDDTAVYYCARDSVSKSYSAPPEFWGQGTWTV

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SSASTKG 143 8ANC91HC QLQLQESGPGLVKPSETLSLTCSVSDGSINSNSYYWAWIR QSPGKGLEWIGSVYYFGGTYYSPSLKSRVTMSVDRSKNQF SLNVSSVTAADTAIYYCARHVRPYDRSGYPERPNWFDPWG RGTLVTVSSASTKG 144 8ANC92HC RVQLVQSGAEVKKPGSSVTVSCKASGGSFSSYAISWVRQA PGQGLEWVGGVKVMFGTVHYSQKVQGRVTITADDSTGTSY LELSGLRSADTAVYYCARNAGAYFYPFDIWGQGTLIIVSS AS TKG 145 8ANC93HC QVQLVQS GAEVKKPGASVKVS CKAS GYT FTRYHIHWVRHA PGQGLEWMGKINPSRASTKYAKKFQDRVTMTRDTSTSTVY MELSSLRGDDTAVYYCGREMGTFTLLGWIDHYDFYPMDV WGQGT PVTVS SAS TKG 146 8ANC9HC QVQLVQSGAEVRKPGSSLKVSCKSSGGTFSRYWNWVRQA PGQGLEWMGG11PIEGIAKYAQKFQDRVTMTADE S KNTVY LDFSSLRSGDTAVYYCARDRGDTRLLDYGDYEDERYYYGM DVWGQGT TVTVS SAS TKG 147 12A10HC SQHLVQSGTQVKKPGASVRVSCQASGYTFTNYILHWWRQA PGQGLEWMGLIKPVFGAVNYARQFQGRIQLTRDIYREIAF LDLSGLRSDDTAVYYCARDESGDDLKWHLHPWGQGTQVIV SPASTKG 148 12A12HC SQQLVQSGTQVKKPGASVRISCQASGYSFTDYVLHWWRQA PGQGLEWMGWIKPVYGARNYARRFQGRINFDRDIYRE TAE MDLSGLRSDDTALYFCARDGSGDDTSWHLDPWGQGTLVIV SAASTKG 149 12A13HC SQQLVQSGTQVKKPGASVRISCQASGYSFTDYVLHWYRQA PGQGLEWMGWIKPVYGARNYARRFQGRINFDRDIYREIAF MDLSGLRSDDTALYFCARDGSGDDTSWYLDPWGQGTLVIV SAASTKG 150 12A16HC SQQLVQSGTQVKKPGASVRISCQASGYTFTDYVLHWWRQA PGQGLEWMGWIKPVYGARNYARRFQGRINFDRDIYRE TAY MDLSGLRSDDTARYFCARDGSGDDTSWHLHPWGQGTLVIV SAASTKG 151 12A17HC SQQLVQSGTQVKKPGASVRVSCQASGYTFMNYIIHWWRQA PGQRLEWMGWINPVFGARNYAHRFQGRINFDRDINRETFQ

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MELTGLRSDDTAVYYCARDGSGDARDWHLDPWGQGTLVIV SSASTKG 152 12A1HC SQHLVQSGTQVKKPGASVRVSCQASGYTFTNYILHWWRQA PGQGLEWMGLIKPVFGAVNYARQFQGRIQLTRDINREIAF LDLSGLRSDDTAVYYCARDESGDDLKWHLHPWGQGTQVIV SPASTKG 153 12A20HC SSASTKG SQQLVQSGTQVKKPGASVRVSCQASGYTFMNYIIHWWRQA 154 12A21HC SQHLVQSGTQVKKPGASVRVSCQASGYTFTNYILHWWRQA PGQGLEWMGLIKPVFGAVNYARQFQGRIQLTRDIYREIAF LDLSGLRSDDTAVYYCARDESGDDLKWHLHPWGQGTQVIV SPASTKG 155 12A22HC SQQLVQSGTQVKKTGASVRVSCQASGYDFTKYLIHWWRQA PGQGLEWMGWMKPVYGATNYAHRFQGRIS FTRDIYREIAF MDLNGLRSDDTAVYFCARDGGGDDRTWLLDAWGQGTLVIV SSASTKG 156 12A23HC SQHLVQSGTQVKKPGASVRVSCQASGYTFTNYILHWWRQA PGQGLEWMGLIKPVFGAVNYARQFQGRIQLTRDINREIAF LDLSGLRSDDTAVYYCARDESGDDLKWHLHPWGQGTQVIV SPASTKG 157 12A27HC SQQLVQSGTQVKKPGASVRISCQASGYTFTDYVLHWWRQA PGQGLEWMGWIKPVYGARNYARRFQGRINFDRDIYREIAF LDLSGLRSDDTARYFCARDGSGDDTSWHLHPWGQGTLVIV SAASTKG 158 12A2HC SQQLVQSGTQVKKPGASVRISCQASGYTFTDYVLHWWRQA PGQGLEWMGWIKPVYGARNYARRFQGRINFDRDIYREIAY MDLSGLRSDDTARYFCARDGSGDDTSWHLHPWGQGTLVIV SAASTKG 159 12A30HC SQQLVQSGTQVKKPGASVRISCQASGYTFTDYVLHWWRQA PGQGLEWMGWIKPVYGARNYARRFQGRINFDRDIYREIAY MDLSGLRSDDTARYFCARDGSGDDTSWHLHPWGQGTLVIV SAASTKG 160 12A37HC SQQLVQSGTQVKKTGASVRVSCQASGYDFTKYLIHWWRQA

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PGQGLEWMGWMKPVYGATNYAHRFQGRIS FTRDIYREIAF MDLNGLRSDDTAVYFCARDGGGDDRTWLLDAWGQGTLVIV SSASTKG 161 12A46HC SQQLVQSGAQVKKPGASVRVSCQASGYTFTNHFLHWWRQA PRQGLEWMGWINPVHGGRNYARRFQGRINFGRDVYQETAY MELSGLRNDDTATYFCARGGGDGRNWHLHPWGQGTLVIVS AASTKG 162 12A4HC SQHLVQSGTQVKKPGASVRVSCQASGYTFTNYILHWWRQA PGQGLEWMGLIKPVFGAVNYARQFQGRIQLTRDIYREIAF LDLSGLRSDDTAVYYCARDESGDDLKWHLHPWGQGTQVIV SPASTKG 163 12A55HC SQQLVQSGAQVKKPGASLRVSCQASGYTFMNYLLHWWRQA PGQGLEWMGWINPVYGAVNYAHRFQGRLTFSRDVYREIAY MDLNGLRSDDTAVYFCARDGSGDDRNWHLDPWGQGTLVIV SSASTKG 164 12A56HC SQQLVQSGTQVKKPGASVRVSCQASGYTFTNYILHWWRQA PGRGLEWMGLIKPVYGAVNYARQFQGRIQLTRDIYREIAF LDLSGLRPDDTAVYYCARDESGYDLNWHLDSWGQGTQVIV SPASTKG 165 12A6HC SQQLVQSGTQVKKPGASVRVSCQASGYTFTDYVLHWWRQA PGQGLEWMGWIKPVYGARNYAHRFQGRINFDRDVYREIAY MDLSGLRSDDTAVYFCARDGSGDATSWHLHPWGQGTLVIV SSASTKG 166 12A7HC SQQLVQSGTQVKKPGASVRVSCQASGYTFMNYIIHWWRQA PGQRLEWMGWINPVFGARNYAHRFQGRINFDRDINRETFQ MELTGLRSDDTAVYYCARDGSGDARDWHLDPWGQGTLVIV SSASTKG 167 12A9HC QVTLVQSGAEVKKPGASVRISCRASGFTFDDYSDYSFIPT TYLIHWFRQAPGQGLEWMAWINSVNGGRNIARQFQGRVTV ARDRSNSIAFLEFSGLRHDDTAVYFCARDRRDDDRAWLLD PWGQGTRVTVS SAS TKG 168 LSSB2339HC QVRLE Q S GAAMRKP GASVT L S C QAS GYN FVKYIVHWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLTSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG

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169 LSSB2351HC QVRLE Q S G TAVRKP GASVTISCQAS GYNFVKFFIHWVRQR PGQGFEWVGMIEPFRGRPWSAGNFQGRLSLSRDVSTETLY MTLNNLRSDDTAVYFCARLEAESDSHSRPIMFDHWGHGSL VTVSSASTKG 170 LSSB2361HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 171 LSSB2364HC QVRLEQSGAAVRKPGASVTLSCQASGYNFVNYIIHWVRQR PGLDFEWVGMIDPYRGRPWSAHKFQGRLSLSRDVSTEILY MTLSSLRSDDTATYFCARAEAESQSHSRPIMFDFWGQGSR VTVSSASTKG 172 LSSB2367HC QVRLSQSGAAIKKPGASVTISCETEGYTFINYIIHWVRQP PGRGFEWLGMIDPRNGRPWFGQSVQGRLSLRRDTYTEWY MTLSGLTSDDAGHYFCARNEPQYHDGNGHSLPGMFDYWGQ GTLVAVSSASTKG 173 LSSB2416HC QVRL S Q S GAAVKKP GASVTIVCE TE GYNFIDY11HWVRQP PGRGFEWLGMIDPRNGRPWSGQKVHGRLSLWRDTSTEKVY MTLTGLTSDDTGLYFCGRNEPQYHDDNGHSLPGMIDYWGQ GTMVTVSSASTKG 174 LSSB2434HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 175 LSSB2483HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 176 LSSB2490HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 177 LSSB2503HC QVRLE Q S GAAVRKP GASVT L S C QAS GYN FVRY11HWVRQR PGLDFEWVGMIDPYRGRPWSAHKFGGRLSLTRDVSTEILY MTLTSLRSDDTATYFCARAEAESQSHSRPIMFDSWGQGSR

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VTVSSASTKG 178 LSSB2525HC QVRLE Q S GNAVRKP GASVTISCQAS GYNFVKFFIHWVRQR PGQGFEWVGMIEPFRGRPWSAGNFQGRLSLSRDVSTETLY MTLNNLRSDDTAVYFCARLEAESDSHSRPIMFDHWGHGSL VTVSSASTKG 179 LSSB2530HC QVRLE Q S GAAMRKP GASVT L S C QAS GYN FVKY11HWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 180 LSSB2538HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFLNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 181 LSSB2554HC QVRLE Q S GAAMRKP GASVT L S C QAS GYN FVKY11HWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 182 LSSB2573HC QVRLSQSGAAIKKPGASVTISCETEGYTFINYIIHWVRQP PGRGFEWLGMIDPRNGRPWFGQSVQGRLSLRRDTYTEWY MTLSGLTSDDTGLYFCARNEPQYHDGNGHSLPGMFDSWGQ GTLVAVSSASTKG 183 LSSB2578HC QVQL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 184 LSSB2586HC QVRLE Q S GAAMRKP GASVT L S C QAS GYN FVKYIVHWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 185 LSSB2609HC QVRL FQS GAAMKKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDISTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 186 LSSB2612HC QVRLE Q S G TAMRKPGASVTL S CQAS GYNFVKYIVHWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY

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MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 187 LSSB2630HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 188 LSSB2640HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 189 LSSB2644HC QVRLSQSGAAIKKPGASVTISCETEGYTFINYIIHWVRQP PGRGFEWLGMIDPRNGRPWFGQSVQGRLSLRRDTYTEWY MTLSGLTSDDTGLYFCARNEPQYHDGNGHSLPGMFDSWGQ GTLVAVSSASTKG 190 LSSB2665HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 191 LSSB2666HC QVRLE Q S GAAMRKP GASVT L S C QAS GYN FVKY11HWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 192 LSSB2669HC QVRLE Q S GAAMRKP GASVT L S C QAS GYN FVKY11HWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 193 LSSB2680HC QVRLE Q S GVAMRKPGASVTL S CQAS GYNFVKY11HWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDIHSRPIILTGPGEYGL DLEHMDWTWRILCLLAVAPGCHSQ 194 LSSB2683HC QVRLE Q S GAAMRKP GASVT L S C QAS GYN FVKYIVHWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 195 LSSB344HC QVRLEQSGTAVRKPGASVTISCQASGYNFVKFFIHGVRQR

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PGQGFEWVGMIEPFRGRPWSAGNFQGRLSLSRDVSTETLY MTLNNLRSDDTAVYFCARLEAESDSHSRPIMFDHWGHGSL VTVSSASTKG 196 LSSNEC107HC QVRLVQSGPQVKTAGASMRVSCEASGYRFLDYIIVWIRQT HGQHFEYVGMINPRGGTPWPSSKFRDRLTLTRDIYTDTFY LGLNNLGSGDTAIYFCARLEADGDDYSPKMFDYWGQGTRI IVSAASTKG 197 LSSNEC108HC QVHT FQS GS SMKKS GASVTIS CEAT GYNIKNYILHWVRQK PGRGFEWVGMIDPINGRPWFGQPFRGRLTLTRDLSTETFY MSLSGLTSDDTATYFCARREADYHDGNGHTLPGMFDFWGP GTLITVSSASTKG 198 LSSNEC109HC QVSLVQSGPQVKTPGASMRVSCETSGYRFLDYIIVWIRQT HGQHFEYVGMINPRGGTPWPSSKFRDRLTMTRDIHTDTFY LGLNNLRSDDTAIYFCARLEADGDDYSPKMFDYWGQGTRI IVSAASTKG 199 LSSNEC110HC QVRLVQSGPQMKTPGASLRLSCEVSGYRFLDYFIVWVRQT GGQGFEYVGMINPRGGRPWSSWKFRDRLSLTRDIETDTFY LGLNNLRSDDTAIYFCARLEADGDNYSPKMVDYWGQGTKI IVSPASTKG 200 LSSNEC116HC QVRL S QS GAAWKT GAS VTIS CE TE GYNFVNY11HWVRRP PGRGFEWLGMIDPRNGHPWFAQTVRGRLSLRRDTFKETVY MTLSGLTSDDTGVYFCARNEPQYHSLPGMFDYWGHGTPVT VSSASTKG 201 LSSNEC117HC QVRLVQSGAQLKKPGASVTVSCEASGYNFVNYIINWVRQT PGRGFEWVGMIDPRRGRPWSAQKFQGRLTLTRDIDSEKLY MHLSGLRGDDTAVYYCARQDSDFHDGHGHTLRGMFDSWGQ GSPVTVSSASTKG 202 LSSNEC118HC QVRLVQSGPQVKTPGASMRISCEASGYRFQDYIIVWIRQT HGQGFEYVGMINPRGGTPWSSSKFRDRLSLTRDIYTDTFY LGLNNLGSDDTAIYFCARLEADGGDYSPKMFDYWGQGTRI IVSAASTKG 203 LSSNEC11HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG

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204 LSSNEC122HC QVRLVQSGPQVKRPGASIRLSCETSGYRFQDYIVAWIRQT RGQRFEFVGMVNPRGGRPWPSSKFRDRVTLTRDIESETFH LGLNDLTSDDTATYFCARLEADGADYSPKMFDFWGQGTKI WSPASTKG 205 LSSNEC123HC QVRLEQSGAAVRKPGASVTLSCQASGYNFVNYIIHWVRQR PGLDFEWVGMIDPYRGRPWSAHKFEGRLSLSRDVSTEVLY MTLSSLRSDDTATYFCARAEAESQSHSRPIMFDYWGQGSR VTVSSASTKG 206 LSSNEC127HC QVRLEQSGAAMRKPGASVTLSCQASGYNFVKYIIHWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 207 LSSNEC18HC QVRLSQSGAAVMKTGASVTISCETEGFNFVNYIIHWVRRP PGRGFEWLGMIDPRNGHPWFAQTVRGRLSLRRDT FNEIVY MTLSGLTTDDTGLYFCARNEPQYHSLPGMFDYWGQGTPVT VSSASTKG 208 LSSNEC24HC QVRLSQSGAAMKKPGASVTISCETEGYTFINYIIHWVRQP PGRGFEWLGMIDPRNGRPWFGQSVQGRLSLRRDTYTEWY MTLSGLTSDDAGLYFCARNEPQYHDGNGHSLPGMFDYWGQ GTLVAVSSASTKG 209 LSSNEC29HC QVRLSQSGAAWKTGASVTISCETEGYTFVNYIIHWVRQS PGRGFEWLGMIDPRNGHPWFGQRLRGRLSLRRDRSTETVF MTLSGLTSDTAIYFCARNEPQYYDGSGHSLPGMFDYWGQ GTRVWSSASTKG 210 LSSNEC2HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 211 LSSNEC33HC QVRLVQSGPQVKTPGASIRLSCEASGYRFLDYFIVWVRQT PGQGFEYVGMINPRGGRPWSSWKFRDRLSLTREIDTDTFY LGLSNLRSDDTAIYFCARLEADGDDYSPKMVDYWGQGTKI IVSAASTKG 212 LSSNEC34HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ

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GSLITVSSASTKG 213 LSSNEC3HC QVRLEQSGAAVRTPGASVTLSCQASGYKFVNYIIHWVRQR PGLAFEWVGMIDPYRGRPWSAHSFEGRLSLSRDVSMEILY MTLTSLRSDDTATYFCARAEAESQSHSRPIISTSGAR 214 LSSNEC46HC QVQ FFQS GS SMKKS GASVTIS CEAT GYNIKNHILHWVRQK PGRGFEWVGMIDPINGRPWFGQAFRGRLTLTRDLSTETFY MSLSGLTSDDTATYFCARREADYHDGNGHTLPGMFDFWGP GTLVTVSSASTKG 215 LSSNEC48HC QVRLSQSGAAWKTGASVTISCETEGYTFVNHI IHWVRQP PGRGFEWLGMIDPRNGHPWFGQRLRGRLSLRRDRSTETVF MTLSGLTSDDIGIYFCARNEPQYFDGSGHSLPGMFDYWGQ GTRVWSSASTKG 216 LSSNEC52HC QVRLSQSGAAWKTGASVTISCETEGYTFVNYI IHWVRQP PGRGFEWLGMIDPRNGHPWFGQRLQGRLSLRRDRSTETVF MTLSGLTSDDTGIYFCARNEPQYYDGSGHSLPGMFDYWGQ GTRVWSSASTKG 217 LSSNEC56HC QVRLVQSGPQVKTPGASMRVSCEASGYRFLDYIIVWIRQT HGQHFEYVGMINPRGGTPWPSSKFRDRLSLTRDIHTDTFY LGLNNLGSDDTAIYFCARLEADGDDYSPKMFDHWGQGTRI IVSAASTKG 218 LSSNEC60HC QVRLE Q S GAAVKKPGASVTISCQAS GYNFVKFFIHWVRQR PGQGFEWVGMIEPYRGRPWSAGNFQGRLSLSRDVSTETLY MTLNNLRSDDTAVYFCARLEAESDSHSRPIMFDHWGHGSL VTVSSASTKG 219 LSSNEC66HC QVRLS QS GAAVMKTGASVTIS SE TEGYNFVNY11HWVRRP PGRGFEWLGMIDPKNGHPWFAQAVRGRLSLRRDTFNEWY MTLSGLTSDDTGLYFCARNEPQYHDGNGHSLPGMFDFWGQ GTLVTVSSASTKG 220 LSSNEC70HC QVRLSQSGAAWKTGASVTISCETEGYTFVNYI IHWVRQP PGRGFEWLGMIDPRNGHPWFGQRFRGRLSLRRDRSTETVF MTLSGLTSDDNGIYFCARNEPQYYDGSGHSLPGMFDYWGQ GTRVWSSASTKG 221 LSSNEC72HC QVRLEQSGAAVRKPGASVTLSCQASGYNFVNYIIHWVRQR PGLDFEWVGMIDPYRGRPWSAHKFQGRLSLSRDVSTEILY MTLSSLRSDDTATYFCARAEAESQSHSRPIMFDFWGQGSR

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VTVSSASTKG 222 LSSNEC7HC QVRLEQSGAAVRKPGASVTLSCQASGYNFVNYIIHWVRQR PGLDFEWVGMIDPYRGRPWSAHKFQGRLSLSRDVSTEILY MTLNSLRSDDTATYFCARAEAESQSHSRPIMFDSWGQGSR VTVSSASTKG 223 LSSNEC82HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 224 LSSNEC89HC QVRLEQSGGALRKPGASVTLSCQASGYNFVKYIIHWVRQR PGLGFEWVGMIDPYRGRPWYAHSFAGRLSLSRDTSTETLY MTLSSLKSDDTATYFCARAEAASDSHSRPIMDWTWRILCL LAWPAST KG 225 LSSNEC8HC QVRL FQS GAAMRKPGASVTIS CEAS GYNFMNY FVHWVRQR PGRGFEWLGMINPRGGRPWSAQSVQGRLTLTRDTSTEMFY MRLDGLRSDDTATYFCARNEADYHDGNGHSLRGMFDYWGQ GSLITVSSASTKG 226 LSSNEC94HC QVRLEQSGAAMRKPGASVTLSCQASGYNFVKYIVHWVRQK PGLGFEWVGMIDPYRGRPWSAHKFQGRLSLSRDTSMEILY MTLTSLKSDDTATYFCARAEAASDSHSRPIMFDHWGQGSR VTVSSASTKG 227 LSSNEC95HC QVRLVQSGPQVKRPGASIRLSCESSGYRFQDYIVAWIRQT RGQGFEFVGMVNPRGGRPWPSSRFRDRVTLTRDIESETFY LGLNDLTSDTATYFCARLEADGSDYSPKMFDFWGQGTKI WS PAS TKG 228 LSSNEC9HC QVRLVQSGAQLKKPGASVTVSCEASGYNFVNYIINWVRQT PGRS FEWVGMIDPRRGRPWSAQKFQGRLTLTRDIDSEKLY MHLSGLRGDDTAVYYCARQDSDFHDGHGHTLRGMFDSWGQ GSPVTVSSASTKG 229 LSSB2055HC QVQLVQSGPELMKPGSSVKVSCRASGDNFLTSTFNWLRQA PGQRLEWMGRFIPSLGLITSAPKFSDRLTITADQATLTAY MELTGLTSEDTALYYCARGLCRGGNCRLGPSGWLDPWGRG TQVTVSSASTKG 230 LSSB2066HC QWLIQS GAEVKRPGS S VKVS CKAS GGS FPITWVRQAPGH GLEWMGGINPFFGTTNYAQKFQGRVSITADESTSTTYLHL

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SDLRSEDTAVYFCARENREKWLVLRSWFAPWGQGTLVTVS SASTKG 231 LSSB2068HC EESGPGLVKPSQTLSLTCSVSGDSVSSGGYFWSWIRQHPT KGLECLGYVYYTGNTYYNPSLKSPPTIEVAMANNQVSLKL GSVTAADTAVYYCARIKRFRGGNYFDTWGHGLLVTVSSAS TKG 232 LSSB2080HC LAQLEQSGGGWKPGGSLRLPCAASGFTFIDYYMAWIRLA PGKGLEWLSYISKNGDYTKYSESLKGRFTISRDNAKNLVI LQLNRLRADDTAIYFCARADGLTYFGELLQYIFDLWGQGA RVIVSSASTKGPSVFPLAPSSKSTSGHASV 233 LSSB2133HC QVQLVQSGAEVKKPGASVKISCKASGYSFRNYAVHWVRQA PGQGLEWMGEINGGNGNTEYSQKSQGRLTITRDISATTAY MELSSLRSDDTAVYYCARVAYVHVVTTRSLDNWGQGTLVT VSSASTKG 234 LSSB2182HC QVQIRQSGPGLVKPLETLSLSCIVFGGSFIAYHWTWIRQA PLKGLEWIGDIDQGGDITYSPSLKSRVTMSVDRSKSQFSL KL S SVTAADAAVYYCVRG P PNRYAVT S FT S GTHRERS S YY FDYWGPGTLVTVS SASTKG 235 LSSB218HC KAPATLSLSPGERATLSCRASQSVGSDLAWYQQKPGQAPR LLIYDASNRATAIPARFSGSGSGTDFTLSISSLEPEDFAV YFCQQRYDKITFGQGTRLEIQRTVAAPSVFIFPPSDEQ 236 LSSB2277HC G 237 LSSB2288HC QVYLVQSGPELKKPGASVKISCKASGYNFPKYAIHWVRQA PGQGLQWMGWINGDNGDARYSQKLQGRVTPSTDTSASWY MELKRLRSEDTAVYYCARALYPWEIGGVPSTMGDDYWGQG TLITVSSASTKG 238 LSSB331HC QVHLQQWGAGLLKPSETLSLTCAVSGGSFSGFFWTWIRQS PGKGLEWIGEVNHSGFTHSNPSLESRATISVAASNTQFSL RLASVTAADTAIYFCALRYFDWSPFRRDTYGTDVWGQGTT VIVSSASTKG 239 LSSNEC101HC QVQLVQSGAELKKPGSSVKVSCKASGGTFNNHTFNWVRQA PGQGLEWMGRTIPILGSRDYAKT FQDRVT11ADKS T S TVY

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LELRRLKSEDTGVYYCATSMYYFDSGGYYRNTDLDKWGQG SLVTVSSASTKG 240 LSSNEC106HC GLDLEHDGHHKEEPRASVTVSCEASGYNFVNYIIHWVRLT PGRGFEWMGMIDPRRGRPWSAQKFQGRLTLTRDIDSERLY MQLSGLRGDDTAVYFCARQEPDFHDGHGHTLRGMFDSWGQ GSPVSVSSASTKG 241 LSSNEC112HC QVQLVQSGAELKKPGSSVKVSCKASGGTFSNYAINWVRQA PGQGFEWMGGIIPLFATPTYAQKFQGRVRITADDSTSTAY MELSSLRSDDTAVYFCARPNWRSALDYWGQGTLVTVSSA STKG 242 LSSNEC115HC QARLDQWGTGLLKPSETLSLKCAVFGVLFTDYNWTWVRQS PGKGLEWIGHLDHRGGGNYNPSLESRVTISLDYSKAQFSL HLKSVTVADTALYYCAGAVKGFWFDEVYNWFGPWSQGTLV TVASASTKG 243 LSSNEC124HC QVQLQESGPGLVKPSGTLSLTCAVSGASISSRNWWTWVRQ PPGKGLEWIGEIYESGATNYNPSLKSRVTISVDKSKNQFS LRLTSVTAADTAVYFCARLMTFGGLIGTLDYWGQGTLVTV LQPPPRAHRYHPRNLLQEHLCARVMP 244 LSSNEC125HC QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTYAISWVRQA PGQGLEWMGG11PS FSMSNYAQDFQGRLTITADE STS SVY MELNSLRSEDTAVYYCARDFPRFHRLVGNYDFWRGTLDRF SYMDLWGRGTAVTVS SAS TKG 245 LSSNEC126HC QVHLVQSGAEAKRPGSSVRVSCRASGGDFSSYTLSWVRQA PGQGIEWMGGWPMLDTVHYAQKFQGRLTLSVDEGTSTAY MELSSLRSEDTAMYYCTRGRQTFRAIWSGPPAVFDIWGQG TLVIVSSASTKG 246 LSSNEC14HC NGGSLRLSCRVSGFGFHLYEMNWVRQAPGKGLEWISSISG SGESTHYSDSITGRFSMSRDEAKDSLYLQMNNLRVEDTAV YYCTRGFSMGDGTGFSFDTWGRGTMVTVSSGLDTVSLAST KGPSVFPLAPCSRSTSDARLS 247 LSSNEC16HC AARLDQWGTGLVKPSETLSLKCAVFGVDFPDYTWTWARQA PGKGLEWIGHRDHRGGSSYNPSLSGRATISLDTSKAQFSL HIKSVTVADTATYYCAGAVAGLWFEDAYNWFGPWSQGTLV TVAAASTKGPSVFPLAPSSKSTSGHASVL 248 LSSNEC21HC QARLDQWGTGLLKPSETLSLKCAVFGVLFTDYNWTWVRQS

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PGKGLEWIGHLDHRGGGNYNPSLESRVTISLDYSKAQFSL HLKSVTVADTALYYCAGAVKGLWFDETYTWFGPWSQGTRV TVASASTKGPSVFPLAPSSKSTSGTRDLS 249 LSSNEC30HC QVQLVQSEAEVKKPGS SVKVS CKAS GGT FRGYTISWVRQA PGQGLEWMGRIIPILGKAIYAPSFQGRVTLTADKSTGTAY MELSRLRSDDTAVYYCAKVKMRGSSGYYYLFDDWGQGTLV TVSSASTKG 250 LSSNEC49HC QVHLVQSGAEVKKPGASVKVSCKVSGYTLSELSIHWVRQG PGRGLEWMANFDPEDGETIYAPQFQGRVTLTEDTSTDTAY MQLTSLRSEDTAVYYCATDRYTDTGRWGPGTLVTVSSAST KG 251 LSSNEC54HC QARLDQWGTGLLKPSETLSLKCAVFGVLFTDYNWTWVRQS PGKGLEWIGHLDHRGGGSYNPSLESRVSISLDYSKAQFSL HLKSVTVADTALYYCAGAVKGFWFDEPSTWFGPWSQGTMV TVASASTKG 252 LSSNEC55HC QARLDQWGTGLLKPSETLSLKCAVFGVLFTDYNWTWVRQS 253 LSSNEC57HC QARLDQWGTGLLKPSETLSLKCAVFGVLFTDYNWTWVRQS PGKELEWIGHLDHRGGGNYNPSLESRVTISLDYSKAQFSL HLKSVTVADTARYYCAGAVKGFWFDDPYTWFGPWSQGTLV TVASASTKG 254 LSSNEC5HC QVHLVQSGAEAKRPGSSVRVSCRASGGDFSSYTLSWVRQA PGQGLERMGGWPMLDTVHYAQKFQGRLTLSVDEGTSTAY MELSSLRSEDTAMYYCTRGRQTFRAIWSGPPWFDIWGQG TLVSVSSASTKG 255 LSSNEC67HC QFRLVQSGPEVKNPGSSVTVSCKASGGTFSGLGINWVRQA PGQGLEWLGDIKTMYGTTNYAPKFQGRVTITADESTSTSY MELSGLRSEDTAVFYCVRELFGHHPAFGVWGQGTSVIVSS ASTKG 256 LSSNEC74HC QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGVSWVRQA PGQGLEWMGWISPYSGNTNYAQRLQDRVTMTTDTSTNTAY MELRSLRSDDTAVYYCAARSYYYYSMDVWGQGTTVTVSSA STKG

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257 LSSNEC77HC QVQLVQSGADVKKPGASVKVSCKVSGYTVSELSIHWVRQA PGKGLEWMGGFDPEDGKTVSAQNFQGRVTMTEDKSTGTAN MELRSLRSEDTAVYYCATTVQLIVDFCNGGPCYNFDDWGQ GTLVTVSSASTKG 258 LSSNEC85HC VSSASTKG 259 LSSNEC91HC AS TKG 260 LSSNEC92HC QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQP PGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSL KLSSVTAADTAVYYCARLPIGSGWYGRDYWGQGTLVTVSS AS TKG 261 3A124HC EVQLLESGGGLVRPGGSLXLSCSASGFTFNSYAMSWVRQA PGKGLEWVS SVSASGEMTYYADSVRGRFTISRDNANNALH LQMNSLRAEXTAVYYCAKVGGTVWSGYSNYLDYWGPGTLV TVSSASTKG 262 3A125HC QVQLVQS GAEVKKPGASVKVS CKPSSNTFTSHYIHWVRQA PGQGLEWMGMINPGGSTRYAPKFQGRVTLTRDTSTRTVYM ELSSLRSEDTAVYYCARPQYNLGRDPLDVWGLGTMVTVSS AS TKG 263 3A140HC EVQLVESGGGLVKPGGSLRLSCADSGFTFRSYSMHWVRQA PGKGLAWVS SISSTSNYIYYADSVKGRFTISRDNAKNS LY LQMNSLRAEDTAVYYCARTFITASWFDSWGQGTLVTVSSA STKG 264 3A144HC VSGGRFSNYGLSWVRQAPGQGLEWMGRIVPAINRAKYAQK FQGRVILTADKITDTAYMELRSLRSEDTAIFYCARDPQIE IRGNAFDIWGQGTWTVSSASTKG 265 3A160HC QVQLQESGPGLVKPSGTLSLTCNVYGGSMISYYWSWIRQP PGKGLEWIGHVYNSGNTKYSPSLKNRVTISMDTSRNLFSL KVTSVTPADTAVYYCARADYDNIWDSRGGFDLWGQGTLVT VSSASTKG

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266 3A18HC FTADRS TI TAYMDLRGLRSDDTAVYYCVRDPQVEVRGNAFDIWGQGTM VTVSSASTKG 267 3A204HC QVQLVQSGAEMKKPGASVKVSCKASGHTFTNYYMHWVRQA PGQGLEWMGMINPTGDSTRYAQRFQGRVTMTRDTSTRTVY MELSSLRSDDTAVYYCARAHHDFWRAPVDVWGKGTTVTVS SASTKG 268 3A228HC EVQLVQS GAEVKKPGE SLRISCKTS GYNFNDDWIAWVRQR PDKGPEWMGIFYPGDSQATYSPSFQGHVTFSADTSISTAY LQWTSLKASDTAIYYCARTRCFGANCFNFMDVWGKGTALT VTVSSASTKG 269 3A233HC QVQLQESGPGPVKPSETLSLTTCTVSGGSMISYYWSWIRQP PGKGLEWIGYIFTNGRTTYSPSLRSRVTISLDTSTNHFSL RLKSVTAADTAIYYCARLDGEAFRYYLDLWGQGNLVTVSS ASTKG 270 3A244HC IRSFYWHWIRQSPGKGLEWLGSVFDNGLTTHNPSLKSRLT ISEDPSRNQISLKLRSMTAADTAVYYCARGDYDILTSSYQ FDYWGQGTLVAVS SASTKG 271 3A255HC QVQLQESGPGLVKPSETLSLTCTVFGASIRSFYWHWIRQS PGKGLEWLGSVFDNGLTTYNPSLKNRLSISEDPSRNQISL NLRSMTAADTAVYYCARADYDLLTSSYHFDSWGQGTLVTV SSASTKG 272 3A296HC ASTKG 273 3A334HC QVQLVQSGAEVKKPGASVKVSCKAPGYTFIGHYMHWIRQA PGQGLEWMGWINPNSGDTNYAQTFQGRVTMTRDTSISTAY MELTRLRSDDTAVYYCARDLRPMRGNWAMHVWGEGTTVTV SSASTKG 274 3A366HC CTVSGGSISSAGYYWTWIRQHPGKGLEFIGYIYYIGTTYY NPSLKSRLTISIDTSKNQFSLKLSSVTAADTAIYYCARDY TARGRHFFDYWGQGALVTVS SASTKG 275 3A381HC SSFAISWVRQAPGQGLEWMGGIIPIFEATSYAQKFQDRLT

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ITTDESTTTAYMDLSSLRSEDTAVYYCARAQGDILTEGYF DYWGQGTLVTVS SASTKG 276 3A384HC QVQLVQSGAEVKKPGSSVKVSCKVSFFSNYGISWVRQRPG QGLEWMGRIIPAIDDMTYAQT FRGRVT FSADKFT T TAYME LTGLTFEDTATYFCARDPQVNRRGNCFDHWGQGTLVTVSS ASTKG 277 3A419HC LEWMGRIIPAIDDVTYAQTFRGRVTFSAADKFTTTAYMDLT GLRSEDTATYFCARDPQVNRRGNCFDHWGQGTLVTVSSAS TKG 278 3A461HC QVQLVQSGAEVKKPGAAVKISCKASRFTFSSYYIHWVRQA PGQGLEWMGIINPSGGSTSNAQKFQDRVTLTRDMSTGTVY MELSRLTSEDTAVYYCATPEPSSIVAPLYYWGQGTLVTVS SASTKG 279 3A474HC EVQLLESGGGLVQPGGSLRLSCAVSGFTFGGHAVSWVRQA PGKGLEWLSQISGTGTGSRTDYADAVKGRFTVSRDNSKKTVY LQMNSLRVEDTALFYCATRSPGGGYAFDIWGQGAMVTVSS 280 3A518HC QVQLQESGPGLVKPSETLSLTCTVSGGSISSAGYYWSWIR QHPEKGLEFIGYIYYLGTTYYNPSLKSRVSISIDTSNNQF SLELSSVSAADTAIYYCARDYTASGRHFFDYWGQGTLVTV SSASTKG 281 3A539HC EVQLLESGGALVQPGGSLRLSCAASGFTFSTSSMSWVRQA PGKGLEWVSAIGSGRGS T FYADSVKGRFTISRDNSKNTLS LQMNSLTAEDTATYYCTKTGGLLRFPEVWGKGTTVTVSSA STKG 282 3A576HC QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYAISWVRQA PGQGLEWMGGIIPIFEAASYAQKFQDRLTITTDESTTTAY MDLSSLRSEDTAIYYCARAQGDILTEGYFDYWGQGTLVTV S SAS TKG 283 3A613HC QVQLQESGPGLVKPSETLSLTCTVSGGSISTYYWSWIRQP PGKGLEWIGYISYSGSTNYNPSLKSRVTISVDTSKNQFSL KLSSVTAADTAVYYCARHKSVLLWFRELDYWGQGTLVTVS SASTKG 284 3A64HC QVQLVQSGAEVKKPGSSVKVSCKTSGVRFSSNAISWVRQA PGQGLEWMGRTTPMLGGANHAPS FKGRVTISADE S TRTVY

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MEMSSLRYEDTAVYYCASGRREGLNFLLDYWGQGTLVTVS SASTKG 285 3A650HC QVQLVQS GAEVRKPGASVKVS CKTSGYTFTNSYIHWVRQA PGQGLEWMGIINPPGGNTYYAQKFHGRVTLTRDTSTSTVY MELNSLRSEDTAVYFCARPHSPTNIPSRPLDYWGQGTLVT VSSASTKG 286 3A67HC QVQLVQSGAEVKKPGASVKVSCKVSGYPLAELSVHWVRQV PGKGLEWVGGFDPEEGKTVYAQKFQGRVTMTEDRSTDTVY MELISLRYEDTAVYYCATDNPVLQLGELSSSLDYWGQGTL VTVSSASTKG 287 3A779HC PSETLSLTCRVSGASISNFYWTWIRQPAGKGLEWIGRLYS SDKTNYNPSLNGRVTMSLDTSKNQFSLRLTSMTDADTAIY YCAREKGQWVTLPPYYFDSWGQGILVTVSSASTKG 288 3A816HC NTFTSHYVHWVRQAPGQGLEWMGMINPGGTTRYAPKFQDR VTLTRDTSTRTVYMELRSLRSEDTAVYYCARPQYNLGREP LNVWGQGTMVTVS SAS TKG 289 3A869HC QVQLQESGPGLVKPSETLSLTCSVSGASISNFYWTWIRQP AGKGLEWVGRLYSSDRTNYNPSLNGRVTMSLDTSKNQFSL RLTSMTDADTAIYFCAREKGQWLTVPPYYFDSWGQGILVT VSSASTKG 290 3A93HC CTVSGGSIISYYWNWIRQSPGKGLEWLGYIFDGGRANYNP SLRSRLTMSVDTSKNQISLKVKSVTAADSAIYYCARLDGE AFRYYFDSWGQGTLVTVSSASTKG 291 3A966HC QTLSLTCSVSGGSISSAGYYWGWIRQHPGKGLEWIGHIYY SGNTNYNPSLKSRLSMSVETSKNQFSLNLASVTAADTAVY FCARDYSAAGRHL FDSWGQGILVTVS SAS TKG 292 3A978HC KPSQTLSLTCTVSGGSISSAGYYWTWIRHHPGKGLEFIGY IYHIGTPYYNPSLKSRLTISIDTSKNQFSLKLSSVTAADT AIYYCARDYTARGRHFFDYWGQGALVTVS SAS TKG 293 3ANC3HC QVQLVQSGADVKKPGASVTVSCKTDEDEDDFRAHLVQWMR QAPGQRLEWVGWIKPQTGQPSYAQKFQGRVTLTREVSTST VFLQLRNLRSDDTAVYYCARPRGGRDNWSFHVWGRGTLVT VSSASTKG 294 3ANC42HC QVQLVQSGAAVKKPGASVKVSCETYGYTFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY

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MELRGLKFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 295 3ANC66HC QVQLVQSGAAVKKPGASVKVSCETYGYKFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY MELRGLRFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 296 3ANC79HC QVQLVQSGAAVKKPGASVKVSCEAYGYKFTDHFMHWWRQA PGQGLEWMGWINPYTSAVNYSPKYQGRVTMTRDTFLETVY MELRGLRVDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 297 3B10HC ASTKG 298 ZV120NS LQQLQVPRLSMWRVFKVAAATGAQTLTVEEPGSSVKVSCK ASGGSSTAYGYSWVRQAPGQGFEWMGRIIPFYGIITYAPK FQGRVTITADRS T S TVYME L T S L T FADTAL FFCARD FGDP RNGYYFDSWDQGLWLTVSSASTKG 299 ZV126NS QVHLVQSGAEVKKPGSSVRVSCKASGWTFGDSVNSAITWV RQAPGQGLEWMGRFIPILGLSNYAQKFQDRVTINVDRSTN TAYMELSGLRSEDTAVYYCARLITGMNAPWFYYMDVWGKG TTITVSSASTKG 300 ZV129NS FICFSVWRLLEFGGRLVQPGGSLRLSCSASGFTFSNSAM SWVRQAPGKGLEWVS SIL S S GVGT FYADSVKGRFTVSRDN SRNTLYLQMKSLRAEDTALYYCAKVQIQQLNFGVITDAGL DVWGKGT TLIVS SASTKG 301 ZV142NS QVQLGQSGTEVKKPGFSVKVSCKASGGSSTAYGYSWVRQA PGQGFEWMGRIIPFYGIITYAPKFQGRVTITADRSTSTVY MELTSLTFADTALFFCARDFGDPRNGYYFDSWDQGLWLTV S SASTKG 302 ZV154NS QVQLVQSGGEVRKPGSSVKVPCKISGNAFSNYGVNWVRQA PGQGLEWVGRIIPVIGVAQHAPKFQGRVTITADKS TTTAY LELSSLRSDDTAVYFCAKDHGDPRTGYYFDYWGQGALVTV SSASTKG 303 ZV165NS QVQLLQSGTEVKKPGSSVKVSCRASGWTLGNSPNSAIGWV

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RQAPGQGLEWIGRIIPILDVTNYAQKFQGRVTISADKS TN IAYMEISSLGSEDTAFYYCARVITGMTSPWYFYMDVWGEG TTVIVSSASTKG 304 3B171HC VQS QVYLVQS GGEVKKPGS SVKVS CKAS GDS FS S SVITWV RQAPGQGPEWMGRIIPVLGVAAYAQNFYGRVTISADTS SN TAYMELSSLRFEDTAVFYCARETGRGGNLALRQYFFDSWG QGTLVTVSSPSTKG 305 3B17HC EVQLVESGGGLVQPGGSLRISCSATGFTFSTHAMHWVRQA PGKGLEYVSAINSNGRSAFYADSVKGRVTISRDNSKNTLF LQMTSLRAEDTAVYYCVKGPLLRYLDSWGQGTLVTVSSAS TKG 306 3B186HC QVQLVESGGGLVKPGGSLRLSCAASGFSFNEYYMSWIRQA PGQGLEWVANIGSSDAYTIYADSVKGRFTISRDNAENTVY LQMNSLRGEDTAVYYCARIEGYCSNSRCSNYFDPWGQGAL VTVSSASTKG 307 3B193HC MFLFLVAGATGVQSQVYLVPFGPEVKKPGSSVKVSCKASG DS FTS SVITWVRQAPGQGPEWMGRVIPVLGVAAYAQKFYG RVTITADTSSNTAYMEVNSLRFEDTAVYYCARETGRGGNL ALRQYFFDSWGQGTLVTVSSPSTKG 308 3B22HC CQVQLVESGGGWQPGRSLRLSCVGSGFTFSSSGMHWVRQ APGKGLEWVAVISSDGSDEYYGDSVEGRFTISRDNSKNTL FLQLDSLEAEDSAVYYCAKTPPHYDALTGYPSSVLEFWGL GTLVTVSSASTKG 309 3B27HC EVQLVESGGGLVQPGGSLRISCSATGFTFSTHAMHWVRQA PGKGLEYVSAINSNGRSAFYADSVKGRVTISRDNSKNTLF LQMTSLRAEDTAVYYCVKGPLLRYLDSWGQGTLVTVSSAS TKG 310 3B29HC QVHLVQSGAEVKKPGSSVRVSCKASGWTFGDSVNSAITWV RQAPGQGLEWMGRFIPILGLSNYAQKFQDRVTINVDRSTN TAYMELSGLRSEDTAVYYCARLITGMNAPWFYYMDVWGKG TTITVSSASTKG 311 3B2HC SGGRLVQPGGSLRLSCSASGFTLSNSAMSWVRQAPGKGLE WVSSILSSGVGTFYADSVKGRFTVSRDNSRNTLYLQMKSL RAEDTALYYCAKVQIQQLNFGVITDAGLDVWGKGTTLIVS SASTKG

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312 3B31HC EVQLVQSGAEVKKPGSSVKVSCKASGGTFTTYDISWVRQA PGQGLEWIGGILPDFGAPSYAQKFQDRVTITTDESSRTAY MELNSLRSEDTAIYYCARGRGDDFWSGESPSWYFDYWGQG TQVTVSSASTKG 313 ZVZZNS PLVQLE P S GVEVKKRGASVKVS CKVS GYS L TE L SMHWVRQ APGKGLEWMSFDPLDGDTIYAQKFQGRVTMTVDTSTDTA YMDLSSLRFEDTAVYYCATPSKAYYYDSPNYEGDFYMDVW GKGTTVIVSSASTKG 314 ZV40NS QVQLVE S GGGWQPGRS LRL S CVGS G FT ES S S GMHWVRQA 315 ZV41NS EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQA PGKGLEWMGVFDPLEGDGVYAEKFRGRVIMTEDTSTDTGY MELTSLRSEDTAIYYCATKAKDYYYESSDYSPYYYYYMDV WGKGT TVTVS SAS TKG 316 ZV44NS EVRLLESGGGLVQPGGSLRLSCSASGFTFSNSALSWVRQA PGKGLEWVSSWSSGGDTFYADSVKGRFTISRDNSRNTLY LQMKSLRAEDTALYYCAKVQIQQLNFGVITDAGMDVWGKG TTVIVSSASTKG 317 ZV45NS VEEPGSSVKVSCKASGGSSTAYGYSWVRQAPGQGFEWMGR IIP FYG11TYAPKFQGRVTITADRS T S TVYMELTRLT FAD TALFFCARDYGDPRNGYYFDSWDQGLWLTVSSASTKG 318 ZV48NS QVQLVESGGGLVQPGGSLRISCSATGFTFSTHAMHWVRQA PGKGLEYVSAINSNGRSAFYADSVKGRVTISRDNSKNTLF LQMTSLRAEDTAVYYCVKGPLLRYLDSWGQGTLVTVSSAS TKG 319 ZV50NS QVQLVQSGPGLVKPSETLSLTCSVSNGSISSGGYYWSWLR QFPGKGLEWIGSIHYTGRTFYNPSLMGRTALSMDTSNNQF SLKVSSVTAADTALYYCARELQWMFVVDPWGQGTLVTVSS AS TKG 320 ZV51NS QVQLLQSGTEVKKPGSSVKVSCRASGWTLGNSPNSAIGWV RQAPGQGLEWIGRIIPILDVTNYAQKFQGRVTISADKS TN IAYMEISSLGSEDTAFYYCARVITGMTSPWYFYMDVWGEG TTVIVSSASTKG

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321 3B56HC QVQLVQSGGEVKKPGASVKVSCKVSGYSLTELSMHWVRQA PGKGLEWMGVFDPLEGDGVYVQKFRGRVIMTEDTSTDTAY MELTSLRSEDTAIYYCATKAKDYYYESSDYSPYYYYYMDV WGKGT TVTVS SAS TKG 322 3B57HC G S EVQLVE S GAEVKKRGASVKVS CKVSGYSLTEL SMHWVR QAPGKGLEWMGSFDPLDGDTIYAQKFQGRVTMTVDTSTDT AYMDLSSLRFEDTAVYYCATPSKAYYYDSPNYEGDFYMDV WGKGTTVIVSSASTKG 323 3B5HC SWQLVESGPGLVKPSETLSLTCSVSNGSISSGGYYWSWL RQFPGKGLEWIGSIHYTGRTMYNPSLMGRPALSMDTSNNQ FSLKLRSVTAADTALYFCARDLQWIFWDPWGQGTLVTVS SASTKG 324 3B61HC SVDERLLEFGGRLVQPGGSLRLSCSASGFTFSNSAMSWVR QAPGKGLEWVS SILS S GVGT FYADSVKGRFTVSRDNSRNT LYLQMKSLRAEDTALYYCAKVQIQQLNFGVITDAGLDVWG KGTTLIVSSASTKG 325 3B6HC QLQLKESGPGMVKPSETLSLTCSVSGASWSANDYWGWIR QAPGKGLECIGIILYTGSTFYNPSLQSRVTISRDPSKNHV SLTLTSVTAADSAVYYCARIPYHSESYYNWIGGFDVWGQ GTRVTVSSASTKG 326 3B77HC QVHLVQSGAEVKKPGSSVRVSCKASGWTFGDSVNSAITWV RQAPGQGLEWMGRFIPILGLSNYAQKFQDRVTINVDRSTN TAYMELSGLRSEDTAVYYCARLITGMNAPWFYYMDVWGKG TTITVSSASTKG 327 3B79HC QVQLGQSGTEVKKPGFSVKVSCKASGGSSTAYGYSWVRQA PGQGFEWMGRIIP FYG11TYAPKFQGRVTITADRS T S TVY MELTSLTFADTALFFCARDFGDPRNGYYFDSWDQGLWLTV SSASTKG 328 3B84HC SQVQLVESGPGLVKPSETLSLTCSVSNGSISSGGYYWSWL RQFPGKGLEWIGSIHYTGRTMYNPSLMGRPALSMDTSNNQ FSLKLSSVTAADTALYFCARDLQWIFWDPWGQGTLVTVS SASTKG 329 3B86HC RVHSQVQLVESGPGLVKPSQTLSLTCTVSGGSISNGGHYW NWIRQHPGKGLEWIGHIYNIATTYYNPSLKSRVSISVDTS KNQFSLKLSSVTAADTAVYYCARGSGRWTIGARIYFDNWG

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QGALVAVSSASTKG 330 3B8HC QVQLVQSGGEVRKPGSSVKVPCKISGNAFSNYGVNWVRQA PGQGLEWVGRIIPVIGVAQHAPKFQGRVTITADKS TTTAY LELSSLRSDDTAVYFCAKDHGDPRTGYYFDYWGQGALVTV SSASTKG 331 3B93HC QVHLVQSGAEVKKPGSSVRVSCEASGWTFGSVNSAITWVR QAPGQGLEWMGRTIPFLGISNYAQKFQGRVTITADKS TNI AYVDVT S L T S QDTAVYYCARLIT GMTAPW FYYMDVWGKGT TVTVSSASTKG 332 3BNC101HC EVQLVQSGSDVKKPGTTVTISCKADEDEDDFTAYNYFMHW VRQAPGQGLEWIGWINPRTGQPNHAKQLQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LIVSSASTKG 333 3BNC124HC QSQVHLVQSGAEVKKPGSSVKVSCQASGGTFNTFAINWVR QAPGQGLEWVGGIIPVFGTASYAQKFQGRVTVTTDESRGT AYMELNSLRSEDTAVYYCARGQTDLNDDLWSDYSTPGFDY WGQGTLVTVS SAS TKG 334 3BNC130HC VSSASTKG 335 3BNC149HC QPQLVQSGSGAEVKKPGASVRISCEASEYNVFDHFMQWVR QAPMEGLEWMGWINPRGGYPSYSPTFQGRLTFTRQPSWDD S TIT FHMELRGLRHDDTAVYYCARPHS PDDAWS LDVWGRG TLVTVSSASTKG 336 3BNC177HC LQPRVHSEVQLVESGAEVKKPGASVKVSCKVSGYTLSDLS MHWVRQAPGKGLEWMGGFDEEDGEITYAQKFQGRVSMTED TSRDTAYMELSSLRSEDTAVYYCATAPRLELGELSSGFHY WGLGTLVTVS SAS TKG 337 3BNC17HC VSSASTKG 338 3BNC48HC IWAPLIAVTFLVLHCESLGTCCCCQASGGTFNTFAINWVR QAPGQGLEWVGGIIPVFGTASYAQKFQGRVTVTTDESRGT

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AYMELNSLRSEDTAVYYCARGQTDLNDDLWSDYSTPGFDY WGQGTLVTVSSASTKG 339 3BNC58HC EVQLVE S GAEVKKPGASVKVS CKVS GYT L S DL SMHWVRQA PGKGLEWMGGFDEEDGEITYAQKFQGRVSMTEDTSRDTAY MELSSLRSEDTAVYYCATAPRLELGELSSGFHYWGLGTLV TVSSASTKG 340 3BNC78HC EVQLVE S GAEVKKPGASVKVACKVS GKKL S DL 341 3BNC82HC CNPRVHS EVQLVE S GAEVKKPGASVKVACKVS GKKL S DL S IHWVRQAPGKGLEWMGGFDEEDGKISYERKFQGRVSMTED TARDTAFMEMSSLRSDDTAVYFCAAAPRLDLGELSSGFHF WGLGTLVTVSSASTKG 342 3BNC8HC EVQLVE S GAEVKKPGASVKVS CKVS GNSLTEFSIHWVRQA PGKGLEWMGGFDPEEGETVPAQKFKGRLTMTEDTSTNTAY MELSSLRSEDTAVYYCSTEPREMGTLTAGFEYWGQGTLVT VSSASTKG 343 3a426hc QVQLQESGPGLVKPSETXSLTCSVSNGSISSGGYYWSWLR QFPGKGLEWIGSIHYTGRTMYNPSLMGRPALSMDTSNNQF SLKLSSVTAADTALYFCARDLQWIFWDPWGQGTLVTVSS ASTKG 344 3a515hc QVQLVQSGAEVKKPGSSVKVSCKASGGTFTTYDISWVRQA PGQGLEWMGGILPDFGAPSYAQKFQDRVTITTDESSSTAY MELNSLRSEDTAIYYCARGRGDDFWSGESPSWYFDYWGQG TLVTVSSASTKG 345 3b46HC GYSEVQLVQSGPGLVKPSQTLSLTCTVSGGSISNGGHYWN WIRQHPGKGLEWIGHIYNIATTYYNPSLKSRVSISVDTSK NQFSLKLSSVTAADTAVYYCARGSGRWTIGARIYFDNWGQ GALVAVSSASTKG 346 3ANC32HC QVQLVQSGADVKKPGATVTVSCKTDEDEDDFRAHLMQWMR QAPGQRLEWVGWIKPQTGQPSYGQKFQGRVTLTREVSTST VFLQLRNLRSDDTAVYYCARPRGGRDNWSFHVWGRGTLVT VSSASTKG 347 3ANC3HC QVQLVQSGADVKKPGASVTVSCKTDEDEDDFRAHLVQWMR

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QAPGQRLEWVGWIKPQTGQPSYAQKFQGRVTLTREVSTST VFLQLRNLRSDDTAVYYCARPRGGRDNWSFHVWGRGTLVT VSSASTKG 348 3ANC41HC QVQLVQSGAAVKKPGASVKVSCETYGYTFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY MELRGLKFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 349 3ANC42HC QVQLVQSGAAVKKPGASVKVSCETYGYTFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY MELRGLKFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 350 3ANC66HC QVQLVQSGAAVKKPGASVKVSCETYGYKFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY MELRGLRFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 351 3ANC70HC QVQLVQSGAAVKKPGASVKVSCETYGYKFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY MELRGLRFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 352 3ANC75HC QVQLVQSGAAVKKPGASVKVSCETYGYTFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY MELRGLKFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 353 3ANC79HC QVQLVQSGAAVKKPGASVKVSCEAYGYKFTDHFMHWWRQA PGQGLEWMGWINPYTSAVNYSPKYQGRVTMTRDTFLETVY MELRGLRVDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 354 3ANC87HC QVQLVQSGGAVKKPGASVKVSCETYGYTFTDHFMHWWRQA PGQGLEWMGWINPYSSAVSYSPRYQGRVTMTRDTFLETVY MELRGLKFDDTAIYYCATPKSGRDYWSFDLWGQGTLVTVS SASTKG 355 3ANC8HC QVQLVQSGADVKKPGASVTVSCKTDEDEDDFRAHLVQWMR QAPGQRLEWVGWIKPQTGQPSYAQKFQGRVTLTREVSTST VFLQLRNLRSDDTAVYYCARPRGGRDNWSFHVWGRGTLVT VSSASTKG

- 66 042696

356 3ANC96HC QVQLVQSGADVKKPGASVTVSCKTDEDEDDFRAHLVQWMR QAPGQRLEWVGWIKPQTGQPSYAQKFQGRVTLTREVSTST VFLQLRNLRSDDTAVYYCARPRGGRDNWSFHVWGRGTLVT VSSASTKG 357 3B106HC QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRHASWDFDT FSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVI VSSASTKG 358 3B16HC QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPCQFQGRVSLTRHASWDFDT FSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTK 359 3B180HC QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPCQFQGRVSLTRQASWDFDT ISFYMDLKALRLDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 360 3B183HC QVRLLQSGAAVTKPGASVRVSCEASGYEIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDS YSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGSQVT VSSASTKG 361 3B191HC QVRLLQSGAAVTKPGASVRVSCEASGYEIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDS YSFYMDLKALRSDDTGVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 362 3B21HC QVRLLQSGAAVTKPGASVRVSCEASGYEIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDS YSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 363 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGPQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 364 3BBM60 QVHLSQSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARQRSDYRDFDVWGSGTQVT

- 67 042696

VSSASTKG 365 3BBM60 QVHLSQSGAAVTKPGASVRVSCEASGYKIRDYSIHWWRQA PGQGLQWVGWINPQTGQPNIPRPFQGRISLTRQASWDFDT FSFYMDLEALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 366 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLEALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 367 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 368 3BBM60 QVHLSQSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDT YSFYMGLKAVRSDDTAIYFCARQRSDFWDFDVWGSGTQVT VSSASTKG 369 3BBM60 QVHLSQSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 370 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISGHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 371 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNIPRQFQGRISLTRQASGDFDT FSFYMDLKALRSDDTAVYFCARQRSDYWDFGVWGSGTQVT VSSASTKG 372 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDIDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 373 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT

- 68 042696

FSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 374 3BBM60 QVHLSHSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 375 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 376 3BBM60 QVHLSQSGAWTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 377 3BBM60 QVHLSQSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARHRSDYWDFDVWGSGTQVT VSSASTKG 378 3BNC101HC EVQLVQSGSDVKKPGTTVTISCKADEDEDDFTAYNYFMHW VRQAPGQGLEWIGWINPRTGQPNHAKQLQGRVTLTRERST STVEMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LIVSSASTKG 379 3BNC102HC QPQLVQSGSGAEVKKPGASVRISCEASEYNVFDHFMQWVR QAPGQGLEWMGWINPRGGYPSYSPRFQGRLTFTRQPSWDD S SVT FHMELRGLRHDDTAVYYCARPHS PDDAWS LDVWGRG TLVTVSSASTKG 380 3BNC104HC EVQLVQS GS DVRKPGATVTVS CKADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 381 3BNC105HC HVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDT FSFYMDLKALRLDDTAIYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 382 3BNC106HC WQLVQS GS DVRKPGAT VT VS CKADE DE DD FTAYNY FMHW

- 69 042696

VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 383 3BNC107HC QVQLVQSGAALKKPGASLRISCQAYGYKFTDYLIHWWRQA PGQGLEWIGWIKPETGQPSYSYKFQGRVSLTRDTFEEILF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGGGSQVLVSSA STKG 384 3BNC108HC QVQLVQSGTAVKKPGASVRVSCQASGYTFTDYFIYWWRQA PGQGLEWLGWINPRTSQPSYPYRFQGRVTLTRDIFEEMLY MDLRGLRSDDTGIYFCARRHSDYCDFDIWGSGTQIIVSSA STKG 385 3BNC10HC EVQLVQSGSDVRKPGATVTVSCKADEDEDDFTAYNYFMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 386 3BNC114HC EVQLVQSGSDVRKPGATVTVSCKADEDEDDFTAYNYFMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 387 3BNC117HC QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRHASWDFDT FSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 388 3BNC126HC QPQLVQSGSGAEVKKPGASVRISCEASEYNVFDHFMQWVR QAPGQGLEWMGWINPRGGYPSYSPTFQGRLTFTRQPSWDD S TIT FHMELRGLGHDDTAVYYCARPHS PDDAWS LDVWGRG TLVTVSSASTKG 389 3BNC127HC EVQLVESGSDVRKPGATVTVSCKADEDEDDFTAYNYFMHW VRQAPGQGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 390 3BNC134HC QVQLVQSGAALKKPGASLRISCQAYGYKFTDHLIYWWRQA PGQGLEWIGWIKPETGQPSYSYKFQGRVSLTRDTFQEILF MNLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQILVSSA STKG

- 70 042696

391 3BNC140HC EVQLVQS GS DVRKPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 392 3BNC141HC WQLVQS GS DVRKPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 393 3BNC142HC QVQLVQSGAALRRPGASVRISCQAYGYRFTDHLIYWWRQA PGQGLEWIGWIRPETGQPSYSYRFQGRVTLTRDTFEEIHF MDLRGLRYDDTATYFCARRHSDYCDFDVWGSGSQVSVSSA STRG 394 3BNC148HC QVQLVQS GS DVRRPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSRGRGTS LTVSSASTRG 395 3BNC149HC QPQLVQSGSGAEVRRPGASVRISCEASEYNVFDHFMQWVR QAPMEGLEWMGWINPRGGYPSYSPTFQGRLTFTRQPSWDD S TIT FHMELRGLRHDDTAVYYCARPHS PDDAWS LDVWGRG TLVTVSSASTRG 396 3BNC151HC QVQLVQSGATLRRPGASVRISCQAYGYRFTDHLIHWWRQA PGQGLEWIGWIRPETGQPSYAYRFQGRVSLTRDTFEEILF MDLRGLRSDDTAIYFCARRHSDYCDLDVWGGGTQLLVSSA STRG 397 3BNC153HC QVQLVQSGAALRRPGASLRISCLTYGYRFTDHLIYWWRQA PGQGLEWIGWIRPETGQPSYSYRFQGRVSLTRDTFEEIVF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVIVSSA STRG 398 3BNC156HC QVQLVQSGAALRRPGASLRISCQTYGYRFTDHLIYWWRQA PGQGLEWIGWIRPETGQPSYSYRFQGRVSLTRDTFEEIVF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGGGSQVIVSSA STRG 399 3BNC158HC QVQLVQSGAALRRPGASLRISCQTYGYRFTDHLIYWWRQA PGQGLEWIGWIRPETGQPSYSYRFQGRVSLTRDTFEEIVF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVIVSSA

- 71 042696

STKG 400 3BNC159HC QVQLVQSGAALRRPGASVRISCQTYGYRFTDHLIHWWRQA PGQGLEWIGWIKPDTGQPSYSSRFQGRVSLTRDTFEEIVF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVLVSSA STKG 401 3BNC15HC QVQLVQSGAALKKPGASLRISCQTYGYKFTDHLIYWWRQA PGQGLEWIGWIKPETGQPSYSYRFQGRVSLTRDTFEEIVF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVLVSSA STKG 402 3BNC173HC QVQLVQS GS DVRKPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 403 3BNC175HC EVQLVQS GS DVRKPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 404 3BNC176HC QVQLLQSGAAVTRPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPRTGQPNNPRQFQGRVSLTRHASWDFDT FSFYMDLRGLRSDDTAIYFCARQRSDYWDFDVWGSGTQVT VSSASTRG 405 3BNC181HC EVQLVQS GS DVRRPGATVTVS GRADE DE DD FTAYDY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 406 3BNC186HC EVQLVQS GS DVRRPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 407 3BNC18HC EVQLVQS GS DVRRPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 408 3BNC193HC QVQLVQSGTAVRRPGASVRVSCQASGYTFTDYFIYWWRQA PGQGLEWLGWINPRTSQPSYPYRFQGRVTLTRDIFEEMLY

- 72 042696

MDLRGLRSDDTGIYFCARRHSDYCDFDIWGSGTQIIVSSA STKG 409 3BNC196HC QVQLLQSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRISLTRQASWDFDT FSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 410 3BNC20HC QVQLVQS GS DVRKPGATVTVS CKADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 411 3BNC29HC WQLVQSGSDVRKPGATVTVSCKADEDEDDFTAYNYFMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 412 3BNC31HC QVQLVQSGAALKKPGASVRISCQTYGYKFTDHLIYWWRQA PGQGLEWIGWIKPETGQPSYSYRFQGRVSLTRDTFEEIVF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVLVSSA STKG 413 3BNC33HC WQLVQSGSDVRKPGATVTVSCKADEDEDDFTAYNYFMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 414 3BNC42HC QVQLVQSGAALKKPGASVRISCQAYGYKFTDYLIHWWRQA PGQGLEWIGWIKPETGQPSYSYKFQGRVTLTRDTFEEILF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVIVSSA STKGA 415 3BNC44HC EVQLVESGSDVRKPGATVTVSCKADEDEDDFTAYNYFMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 416 3BNC45HC WQLVQSGSDVRKPGATVTVSCKADEDEDDFTAYNYFMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 417 3BNC53HC QVQLVQSGAALKKPGASVRISCQAYGYKFTDHLIYWWRQA

- 73 042696

PGQGLEWIGWIKPETGQPSYAYKFQGRVTLTRDTFEEIHF MDLRGVRNDDTATYFCARRHSDYCDFDVWGSGSQVIVSSA STKG 418 3BNC54HC EVQLVE S GS DVRKPGATVTVS CKADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 419 3BNC55HC QVQLVQSGTAVKRPGASVRVSCQASGYTFTDYFIYWWRQA PGQGLEWLGWINPLTSQPSYPSRFQGRLTLTRDTFDEMLY MDLRGLRSDDTGIYFCARRHSDYCDFDIWGSGTQIIVSSA STKG 420 3BNC59HC EVQLVQS GS DVRKPGATVTVS CKADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMKLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 421 3BNC60HC QVHLSQSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDT YSFYMDLKAVRSDDTAIYFCARQRSDFWDFDVWGSGTQVT VSSASTKG 422 3BNC62HC QVRLLQSGAAVTKPGASVRVSCEASGYEIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDS YSFYMDLKALRSDDTGVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 423 3BNC64HC QVHLSQSGAAVTKPGASVRVSCEASGYKISDHFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDT YSFYMDLKALRSDDTAIYFCARQRSDFWDFDVWGSGTQVT VSSASTKG 424 3BNC65HC QVQLL P FGGAVTKPGASVRVS CEAS GYNIRDY FIHWWRQA PGQGLQWVGWINPKTGQPNNPCQFQGRVSLTRPASWDFDT ISFYMDLKALRLDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 425 3BNC66HC QVQLVQSGAALKKPGASLRISCQTYGYKFTDHLIYWWRQA PGQGLEWIGWIKPETGQPSYSYRFQGRVSLTRDTFEEIAF MDLRGLRSDDTAIYFCARRHTDYCVFDVWGSGSQIIVSSA STKG

- 74 042696

426 3BNC6HC QVQLVE S GS DVRKPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHAKQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTKG 427 3BNC72HC QVQLVQSGAALKKPGASLRISCQTYGYKFTDHLIYWWRQA PGQGLEWMGWIKPETGQPSYSYRFQGRVSLTRDTFEEIVF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVIVSSA STKG 428 3BNC75HC QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRHASWDFDT FSFYMDLKALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VYSASTKG 429 3BNC79HC QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRQASWDFDT ISFYMDLKALRLDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTKG 430 3BNC81HC RQVQLVQSGAALKKPGASLRISCQAYGYKFTDHLIYWWRQ APGQGLEWIGWIKPETGQPSYSYKFQGRVSLTRDTFQEIL FMDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQILVSS AS TKG 431 3BNC84HC QVQLVQSGAALKKPGASLRISCQAYGYKFTDHLIYWWRQA PGQGLEWIGWIKPETGQPSYSYKFQGRVSLTRDTFQEILF MDLRGLRSDDTAIYFCARRHSDYCDFDVWGSGSQVIVSSA STKG 432 3BNC86HC QVQLVQS GS DVRKPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 433 3BNC87HC QVQLLQSGAAVTRPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPRTGQPNNPRQFQGRVSLTRHASWDFDT FSFYMDLRALRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTRG 434 3BNC89HC QVQLVQ S G TAVRRP GAS VRVS C QAS GYTFIDHFIYWWRQA PGQGLEWLGWINPLTSQPSYPSRFQGRLTLTRDTFDEMLY MDLRGLRSDDTGIYFCARRHSDYCDFDIWGSGTQIIVSSA STRG 435 3BNC91HC QVQLLQSGAWTRPGASVRVSCEASGYRIRDYFIHWWRQA PGQGLQWVGWINPQTGQPNIPRPFQGRVTLTRHASWDFDT FSFYMDLRALRSDDTAIYFCARRRSDYCDFDVWGSGTHVT VSSASTRG 436 3BNC92HC EVQLVQSGSDVRRPGATVTVSCRADEDEDDFTAYNYFMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 437 3BNC94HC QVQLVQS GS DVRRPGATVTVS GRADE DE DD FTAYNY FMHW VRQAPGHGLEWIGWINPRTGQPNHARQFQGRVTLTRERST STVFMRLTNLRLDDTAVYFCARPLRGGDTWHYHSWGRGTS LTVSSASTRG 438 3BNC95HC QVQLLQSGAAVTRPGASVRVSCEASGYNIRDYFIHWWRQA PGQGLQWVGWINPRTGQPNNPRLFQGRVSLTRHASWDFDT FSFYMDLRAVRSDDTAVYFCARQRSDYWDFDVWGSGTQVT VSSASTRG

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Table B

SSEQ ID NO: Antibody Amino acid sequence of a light chains 439 8ANC131KC EIVLTQSPATLSLSPGERATLSCRASQGLNFWWYQQKRGQ APRLLIHAPSGRAPGVPDRFSARGSGTEFSLVISSVEPDDF AIYYCQEYSSTPYNFGPGTRVDRKRTVAAPSVFIFPPSDEQ 440 8ANC134KC EIVLTQSPATLSLSPGERATLSCRASQGLNFWWYQQKGGQ APRLLIHGPTDRAPGVPDRFSARGSGTEFSLVISSVEPDDF ALYYCQEYSSTPYNFGPGTRVDRKRTVAAPSVFIFPPSDEQ 441 8ANC13KC EIVLTQSPATLSLSPGERATLSCRASQGLNFWWYQQKRGQ APRLLIHGPSHRAPGVPDRFSARGSGTEFSLVISSVEPDDF AIYYCQEYSSTPYNFGPGTRVDRKRTVAAPSVFIFPPSDEQ 442 8ANC45KC EIVLTQSPATLSLSPGERATLSCRASQGVNFWWYQQKRGQ APRLLIYGPSNRAPGVPDRFSARGSGTEFSLVISSVEPDDF ALYYCQEYSSTPYNFGPGTRVDRKRTVAAPSVFIFPPSDEQ 443 8ANC50KC EIVLTQSPTTLSLSPGERATLSCRASQGVNLWWYQQKRGQ APRLLIYGPSDRAPGVPDRFSARGSGTEFSLVISSVEPDDF ALYYCQEYSSTPYNFGTGTRVDRKRTVAAPSVFIFPPSDEQ 444 8ANC88KC EIVLTQSPATLSLSPGERATLSCRASQGLNFWWYQQKRGQ APRLLIHAPSDRAPGVPDRFSARGSGTDFSLVISSVEPDDF AIYYCQEYSSTPYNFGPGTRVDRKRTVAAPSVFIFPPSDEQ 445 8ancl82kc EIVLTQSPATLSLSPGERATLSCRASQGVNFWWYQQKRGQ APRLLIYGPSDRAPGVPDRFSARGSGTEFSLVISSVEPDDF ALYYCQEYS S T PYNFGTGTRVDRKRTVAAP 446 8ancl92kc EIVLTQSPATLSLSPGERATLSCRASQGVNFWWYQQKRGQ APRLLIYGNSDRVPGVPDRFSARGSGTEFSLVISSVEPDDF ALYYCQEYSSTPYNFGPGTRVDRKRTVAA 447 8ANC14KC SEIVLTQSPATLSLSPGERATLSCRASQSINNYLAWYQQKP GQAPRLLIYDASNRATGIPARFSGGGSGTDFTLTISSLEPE DFAVYYCQQRANWRLLTFGGGTKVEIKRTVAAPSVFIFPPS DEQ 448 8ANC16KC EIVMTQSPDTLSVSPGERATLSCRASQSVNSNLAWYQQKPG QAPRLLIYGASTRATAVPARFSGSGSGTEFTLTISSLQSED SAVYYCQQYYQWLSYTFGQGTKLEIKRTVAAPSVFIFPPSD EQ 449 8ANC195KC EQ 450 8ANC24KC SEIVMTQSPATLSMSPGERATLSCRASLSVNTNLAWYQQKP GQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSE DFALYYCQQYNHWPQTFGQGTKVEIKRTVAAPSVFIFPPSD EQK 451 8ANC5KC DIQMTQSPPSLSASVGDRVTITCQASQDINNFLNWYQQKPG KAPRLLIYDASNLESGVSSRFSGSRSGTDFTLTISSLLPED IATYSCQQYSNLPYTFSQGTKLEIKRTVAAPSVFIFPPSDE Q 452 12a12x DIQMTQSPSSLSASVGDRVTITCQAGQGIGSSLQWYQQKPG KAPKLLVHGASNLHRGVPSRFSGSGFHTTFSLTISGLQRDD FATYFCAVLEFFGPGTKVEIKRTVAAPSVFIFPPSDEQLKS 453 12al3kc DIQMTQSPSSLSASVGDRVTITCQAGQGIGSSLQWYQQKPG

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KAPKLLVHGASNLHRGVPSRFSGSGFHTTFSLTISGLQRDD FATYFCAWEFFGPGTKVDIKRTVAAPSVFIFPPSDEQL 454 12a16x DIQMTQSPSSLSASVGDRVTITCQASQGIGSSLQWYQQKPG RAPNLLVHGASKLHRGVPSRFSGSGFHTTFSLTISGLQRDD FATYFCAVLEFFGPGTKVEIKRTVAAPSVFIFPPSDEQLK 455 12alkc DIQMTQSPSSLSASVGDRVSINCQAGQGLGSSLNWYQQKPG RAPKLLVHGASNLQRGVPSRFSGSGFHTTFTLTISSLQPDD VATYFCAAFQWFGPGTKVEIKRT 456 12a20kc DIQMTQSPSSLSASVGDRVSIHCQAGQGIGSSLNWYQQKPG RAPRLLVHGASNLQRGVPSRFSGSGFHTTFTLTISSLQPDD VATYWCAALEFFGPGTKVEI 457 12a21kc DIQMTQSPSSLSASVGDRVTINCQAGQGIGSSLNWYQKKPG RAPKLLVHGASNLQRGVPSRFSGSGFHTTFTLTISSLQPDD VATYFCAVFQWFGPGTKVDIKRTVAAPSVFIFPPSDEQLK 458 12a22kc DIQMTQSPSSLSASVGDRVTITCQAGQGIGSSLNWYQQKPG RAPKLLVYGASNLQRGVPSRFSGSGFHTTFTLTISSLQPED FATYFCSVYEFLGPGTKVEIKRTVAAPSVFIFPPSDEQ 459 12a23kc DIQMTQSPSSLSVSVGDRVSITCRATQGIGNSLNWYQQKPG KAPKVLIYGTTKLHGGVPSRFSGGGSGSTGTLTIDSLQPED IATYFCQLFEFFGPGTKVEIKRTVAAPSVFIFPPSDEQ 460 12a27kc DIQMTQSPSSLSASVGDRVTITCQASQGIGSSLQWYQQKPG RAPNLLVHGASNLHRGVPSRFSGSGFHTTFSLTISGLQRDD FATYFCAVLEFFGPGTKVDIKRTVAAPSVFIFPPSDEQ 461 12a4 6kc DIQMTQSPSSLPASVGDTVTITCQAGQGIGSSLQWYQQRPG RAPNLLVYDASNLQRGVPSRFTGTGFHTTFTLTIRGLRPED FGTYFCASLEFFGPGTKVDIKRTVAAPSVFIFPPSDEQ 462 12a55kc YIQMTQSPSSLSASIGDRVTITCQAGQGIGSSLNWYQQKPG KAPKLLVHGASNLQRGVSSRFSGSGFHTTFTLTISSLRPED VGTYFCEVYEFIGPGTKVDIKRTVAAPSVFIFPPSDEQ 463 12a56kc DIQMTQSPSSLSASVGDRVSINCQAGQGIGSSLNWYQQKRG KAPKLLVHGASTLQRGVPSRFSGSGFHTTFTLTISSLQPDD VATYFCESFQWFGPGTKVEIKRTVAAPSVFIFPPSDEQ 464 12a6kc DIQMTQSPSSLSASVGDRVTITCQASQGIGSSLQWYQQKPG RAPKLLVHGASNLHRGVPSRFSGSGFHTSFTLTISSLQPDD VATYFCAVLEFFGPGTKVEIKRTVAAPSVFIFPPSDEQ

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465 12a7kc DIQMTQSPSSLSASVGDRVSIHCQAGQGIGSSLKWYQQKSG RAPRLLVHGASNLQRGVPSRFSGSGFHTTFTLTISSLQPDD VATYWCAVLEFFGPGTKVEIKRTVAAPSVFIFPPSDEQ 466 LSSB2339LC QSVLTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPNLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEAFYFCATYDSDGSVRLFGGGTTLTVLSQPKAAPSVTLFP PSNGGR 467 LSSB2351LC QSALTQTPSVSGAPGQRVTISCSGGPSNVGGNYVYWYQQFP GAAPKLLIRRDDQRPSGVPDRFSGSKSGNSASLAISGLRLD DEAYYFCATYDSGWSIRLFGGGTRLTVLSQPKAAPSVTLFP PSSEEL 468 LSSB2364LC PP SQAWTQPPSVSGAPGQRVTISCSGGPSNVGGNLVYWYKQF 469 LSSB2367LC QTWTQPPSASGTPGQRVTISCSGGGSNIGGNLVSWYQHFP GAAPKLLIYRNDQRPSGVPDRFSGSKSGTSASLTISGLRSD DEATYFCAAYDCTLSLRLFGGGTTLNVLSQPKAAPSVTLFP PSSEEL 470 LSSB2490LC QSALTQPPSVSGTPGQNVTISCSGGGSNVGGNLVSWYQHFP GAAPKLLIHRDNQRPSGVPDRFSVLKSGNSASLAISGPRSD DEAFYFCAVYDSSLSLGLFGGGTKLTVLSQPKAAPSVTLFP PSSEEL 471 LSSB2530LC QSALTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPTLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEGFYFCATYDSDGSIRLFGGGTALTVLSQPKAAPSVTLFP PSSEELK 472 LSSB2554LC NFMLTQAPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQYP GTAPKLLILRDDQRPSGVPDRFSASKSGNSASLAISELRPD DEAFYFCATYDSDGSIRLFGGGTALTVLSQPKAAPSV 473 LSSB2586LC NFMLTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPNLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEAFYFCATYDSDGSIRLFGGGTTLTVLSQPKAAPSVTLFP P 474 LSSB2612LC QSVLTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP

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GTAPKLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEAFYFCATYDSDGSIRLFGGGTALTVLSQPKAAPS 475 LSSB2640LC QLVLTQPPSVSGTPGQNVTISCSGGGSHVGGNLVSWYQHFP GAAPKLLIHRDNQRPSGVPDRFSALKSGNSASLAISGLRSD DEAFYFCAVYDSSLSLGLFGGGTKLTVLSQPKAAPSVT 476 LSSB2644LC RTWTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQL PGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQA EDEADYYCQSYDSSLSGSGVFGTGTKVTVLGQPKANPTVTL FPPSSEEL 477 LSSB2666LC QSALTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPKLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEALYFCATYDSDGSIRLFGGGTALTVLSQPKAAPSVTLFP PGWEE 478 LSSB2680LC QPVLTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPNLLILRDDQRPSGVPDRFSASKSGNSASLAITGLRPD DEAFYFCATYDSDGSIRLFGGGTALTVLSQPKAAPSVTLFP P 479 LSSB2683LC QSALTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPNLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEAFYFCATYDSDGSIRLFGGGTTLTVLSQPKAAPSVTLF 480 LSSB344LC QSALTQTPSVSGAPGQRVTISCSGGPSNVGGNYVYWYQQFP GAAPKLLIRRDDQRPSGVPDRFSGSKSGNSASLAISGLRLD DEAYYFCATYDSGWSIRLFGGGTRLTVLSQPKAAPSVTLFP PSSEEL 481 LSSNEC107LC QLVLTQPPSVSATPGQTVTISCSGSGSNVGGNHVYWYRQLP GAAPTLVISKTDHRPSRVPDRFSGSKSGNSASLAISGLRPD DEAAYFCATYDTGLSLRLFGGGTRLAVLSQPKAAPSVTLFP PSSEEL 482 LSSNEC108LC QSALTQPPATSGTPGQRVTISCSGGGSNVGGNLVSWYQQFP GAAPKLILHRDGQRPSGVPDRFSASKSGTSASLTISGLRSD DEATYFCAAFDSALSLPLFGGGTKLTVLSQPKAAPSVTLFP PSSEEL 483 LSSNEC117LC QSVLTQVLSVSGTPGQRVIISCSGTSSNVGGNLVSWYQHLP GAAPRLLIHRDDQRPSGVPDRFSGSKSGNSASLVISGLRSD DEADYFCGAYDSTFSLPVFGGGTRLTVLSQPKAAPSVTLFP

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PSSEEL 484 LSSNEC118LC NFMLTQPPSVSATPGQTVTISCSGSGSNVGGNHVYWYRQLP GAAPTLVISKTDHRPSRVPDRFSGSKSGNSASLAISGLRPD DEAVYFCATYDTGLSLRLFGGGTRLTVLSQPKAAPSVTQFP PSSEE 485 LSSNEC122LC QSALTQPPSVSATPGQTVTISCSGSGSNVGGNHVYWYRQLP GAAPTLLISKTNHRPSQVPDRFSASKSGNSASLAISGLRPD DEADYFCGTYDTSLSLRLFGGGTRLTVLSQPKAAPSVTLFP PSSEEL 486 LSSNEC24LC QSALTQPPSASGTPGQRVTISCSGGGSNIGGNLVSWYQHFP GTAPKLLIYRNDQRPSGVPDRFSGSKSGTSASLTISGLRSD DEATYFCAAYDSSLSLRLFGGGTTLNVLSQPKAAPSVTLFP PSSEEL 487 LSSNEC2LC QSALTQPPSVSGTPGQNVTISCSGGGSDVGGNLVSWYQHFP GAAPKLLIHRDNQRPSGVPDRFSALKSGNSASLAISGLRSD DEAFYFCAVYDSSLSLGLFGGGTKLTVLSQPKAAPSVTLFP PSSEEL 488 LSSNEC33LC QAVVTQPPSVSATPGQTVTISCSGSGSNVGGNHVYWYRQLP GAAPTLLISKTNRRPSQVPDRFSGSKSGNSASLAISGLRPD DEADYFCATYDTDLSLRLFGGGTRLTVLSQPKAAPSVTLFP PSSEEL 489 LSSNEC46LC QSALTQPPAASGAPGQRVTISCSGGGSNVGGNLVSWYQQFP GAAPKLILHRDGQRPSGVPDRFSASKSGTSASLTISGLRSD DEATYFCAAYDSAVSLPVFGGGTKLTVLSQPKAAPLVT 490 LSSNEC48LC NFMLTQPPSASGTPGQRVTISCSGGGSNIGGNLVSWYQHFP GAAPKLLIYRNDQRPSGVPDRFSGSKSGTSASLAISGLRSD DKATYFCAAYDSTLSLRLFGGGTTLTVLSQPKAAPSVTLFP PSSEE 491 LSSNEC52LC QSVLTQVLSVSGTPGQRVIISCSGTSSNVGGNLVSWYQHLP GAAPRLLIHRDDQRPSGVPDRFSGSKSGNSASLVISGLRSD DEADYFCAAYDSTFSLPVFGGGTRLTVLSQPKAAPSVTLFP PSSE 492 LSSNEC56LC QSALTQPPSVSATPGQTVTISCSGSGSNVGGNHVYWYRQLP GAAPTLLISKTDHRPSRVPDRFSASKSGNSASLAISGLRPD DEAIYFCATYDTGLSLRLFGGGTRLTVLSQPKAAPSVTLFP

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PSSEEL 493 LSSNEC60LC QSALTRTPSVSGAPGQRVTISCSGGPSNVGGNYVYWYQQFP GAAPKLLIRRDDQRPSGVPDRFSGSKSGNSASLAISGLRLD DEAYYFCATYDSGWSIRLFGGGTRLTVLSQPKAAPSVTLFP PSSEEL 494 LSSNEC70LC QSALTQAPSASGTPGQRVTISCSGGGSNIGGNLVSWYQHFP GAAPKLLIYRNDQRPSGVPDRFSASKSGTSASLAISGLRSD DEATYFCAAYDSTLSLRLFGGGTTLAVLSQPKA 495 LSSNEC72LC NFMLTQPPSVSGAPGQRVTISCSGGPSNVGGNLVYWYKQFP GTAPKLLIRRDDQRPSGVPDRFSGSKSGNSASLAISGLRPD DEAFYFCATYDSHGSIRLFGGGTLLTVLSQPKAAPSVTLFP PSSEEL 496 LSSNEC7LC QLVLTQPPSVSGAPGQRVTISCSGGPSNVGGNLVYWYKQFP GTAPKLLIRRDDQRPSGVPDRFSGSKSGNSASLTISGLRPD DEAFYFCATYDSQGSTRLFGGGTVLTVLSQPKAAPSVTLFP PSSEEL 497 LSSNEC89LC QSALTQPPSVSGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPKLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEAFYFCATYDSQGSFRVFGGGTALTVLSQPKAAPSVTLYP PSSEE 498 LSSNEC94LC NFMLTQPPSASGAPGQRVTISCSGGPSNVGGNYVYWYRQFP GTAPNLLILRDDQRPSGVPDRFSASKSGNSASLAISGLRPD DEAFYFCATYDSDGSIRLFGGGTTLTVLSQPKAAPSVTLFP PSSEEL 499 LSSNEC9LC QVLSVSGTPGQRVIISCSGTSSNVGGNLVSWYQHLPGAAPR LLIHRDDQRPSGVPDRFSGSKSGNASLVISGLRSDDEADY FCAAYDSTFSLPVFGGGTRLTVLSQPKAAPSVTLYAPSSEE 500 LSSB2066KC PVTLSASVGDRVTITCRASEDISKYLNWYQHKPGKAPKLLI YTASSLETGVPSRFSGSGSGTDFSLTISSLQPDDFATYYCQ QSYTSSVTFGQGTRVEVKRTVAAPSVFIFPPSDEQ 501 LSSB2080KC L 502 LSSB2133KC TLSASVGDRVTITCRASQSINNYLNWYQQKPGKAPKLLIYA

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ASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFVTYYCQQT YSNPRMFGQGTKVEIKRTVAAPSVFIFPPSDEQ 503 LSSB2182KC KAPATLSLSPGERATLSCRASQSVGSDLAWYQQKPGQAPRL LIYDASNRATAIPARFSGSGSGTDFTLSISSLEPEDFAVYF CQQRYDKITFGQGTRLEIQRTVAAPSVFIFPPSDEQ 504 LSSB331KC EQL 505 3A124KC SEIVLTQSPATLSLSPGESATLSCRASQSLSSSLAWYQQKP GQAPRLLIYDTSDRATGIPARFSGRGSGTDFTLTISSLEPE DFAVYYCQQRSNWAITFGQGTRLEIKRTVAAPSVFIFPPSD 506 3A125KC EIVLTQSPGTLSLSPGEXATLSCRASQTISNNYLXWYQQKA GQAPRLLIYGASSGATGIPDRFSGSGSGTDFTLTISRLEPE DFAVYYCQQYGLSPWTFGRGTKVEIKRTVAAPSVFIFPPSD 507 3A140LC QSALTQPRSVSGSPGQSVTISCTGTSSDVGAYNYVSWYRQH PGKAPKLMINDVSKRPSGVPDRFSGSKSGNTASLTISGLQA EDEADYYCCSYAGTYSYVFGTGTKVTVLGQPKANPTVTLFP PSSEEL 508 3A144KC APVTLSASVGDTVTITCRASQPIATFLNWYQHKPGQAPKLL IYAASTFQRGAPSRYSGSGSGTDFTLTINSLQPEDLATYYC QQTFTDPVTFGQGTRLEIKRTVAAPSVFIFPPSD 509 3A160KC DIQMTQSPASLSASVGDRVTITCRASQGISHYLAWYQQKPG KVPRLLIYAASRLQSGVTSRFSGSGSGTEFTLTISSLLPED AAVYFCQKYDTDPMTFGQGTRLEIKRTVAAPSVFIFPPSD 510 3A18KC DIQMTQSPSSLSASIGDRVTITCRANQHIRSFLNWYQQTPG KAPKLLIYAASTLQRGVPSRFSGSGSGTDFTLTITSLERED LATYYCQQTYTSPITFGQGTRLEIKRTVAAPSVFIFPPSDE 511 3A204KC EIVLTQSPGTLSLSPGERATLSCRASQSVSNNYLAWYQQKP GQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPE DFAVYYCQQYATSSLYTFGQGTKLEIKRTVAAPSVFIFPPS D 512 3A228KC LSVSLGERATINCKSSQSILYSSDKKNYLAWYQQKIGQPPK LLLYWASTRESGIPDRFSGSGSGSDFTLTISSLQPEDVAVY YCQQYYISPFTFGPGTKVDLKRTVAAPSVFIFPPSD

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513 3A233LC NFMLTQPASVSGSPGQSITLSCTGTTSDVRDSNFVSWYQQV PGKAPKLIIYDVSARPSGVSFRFSGSKSGNTASLTISGLQA EDEALYYCSSFTPTNTLVFGGGTKLTVLGQPKAAPSVT 514 3A244LC SQSVVTQEPSLTVSPGGTVTLTCGPSTGAVTSGFYPHWFQQ KPGQAPRALIYSTSNKYSWTPARFSGSLLGGKAVLTLSDVQ PDDEAEYYCLLLLYYGGPWIFGGGTKLTVLVS 515 3A255LC QAWTQEPSLTVSPGGTVTLTCASSTGAVTSGFYPHWFQQK PGQAPRALIYSTSNRYSWTPARFSGSLLGGKAALTLSGVQP EDEAEYYCLLLPYYGGPWIFGGGTKLTVLGQPKAAPSVTLF PPSSEEL 516 3A296KC EIVMTQSPATLSVSPGDRATLSCRASQSVSTNLAWYQQKPG QAPRLLIYGASTRATGIPATFSGSGFATEFTLTISSLQSED FAVYYCQQYNNWPPAFGQGTKVEIKRTVAAPSVFIFPPSD 517 3A334LC QSVLTQPPSASGSPGQSITISCTGTSSDVGGYNYVSWYQQP PGKAPKVIIYEVSKRPSGVPDRFSGSKSGNTASLTVSGLQA EDEADYYCSSYAGSNNFVFGTGTEVTWGQPKANPTVTLFP PSSEELL 518 3A366KC S L SASVGDRVTITCRASE SIS FYLNWYQQKPGKAPELLIFA TSTLHSGVPSRFSGSGSGTDFTLTISSLQLEDFATYYCQQS SSTPFTFGGGTKVEIKRTVAAPSVFIFPPSD 519 ZAZ84KS DIQMTQSPSSLSAYVGDRVTITCRASQNINTYLNWYQQRPG KAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISNLETED FAVYYCQQTYRSVTFGQGTKLEIKRTVAAPSVFIFPPSD 520 ZA419KS LSAYVGDRVTITCRASQNINTYLNWYQQRPGKAPKLLIYAA STLQSGVPSRFSGSGSGTDFTLTISNLETEDFAVYYCQQTY SSVTFGQGTKLETRRTVAAPSVFIFPPSD 521 ZA461KS SEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQK PVQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEP EDFAVYYCQQYGTLHPRTFGQGTKVEIKRTVAAPSVFIFPP SD 522 ZA474KS EIVLTQSPGTLSLSPGERATLSCRASQSISSNYLAWYQQKP GQAPRLLIYGASTRATGIPDRFSGSGTDFTLSISRLEPE DIAVYYCHQYGSSQRFGQGTKVEIKRTVAAPSVFIFPPSD 523 ZA518KS DIQMTQS PS SLSASVGDRVTITCRASQSISRYLNWYQQKPG KAPKLLIYAASSLQGGVPSRFSGSGSGTDFTLTISSLQPED

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FATYYCQQSSSKPFTFGGGTKVEIKRTVAAPSVFIFPPSD 524 3A539LC NFMLTQPASVSGSPGQSITISCSGTGSDIGVYNYVSWYQQH PGKAPRLMIYDVTNRPSGVSNRFSGSKSGFTASLTISGLQG DDEADYYCSSYSSTNTYVFGTGTHVTVLGQPKANPTVTLFP PSSEEL 525 3A576LC QSALTQPPSASGTPGQRVTISCSGSYHNIGSNAVNWYQQLP GTAPKLLIYSNDQRPSGVPDRFSGSKSGTSASLAISGLQSE DEADYYCAAWDDSLHVFGTGTKVTVLGQPKANPTVTLFPPS SEEL 526 3A613LC QSALTQPPSASGTPGQRVTISCSGSYHNIGSNAVNWYQQLP GTAPKLLIYSNDQRPSGVPDRFSGSKSGTSASLAISGLQSE DEADYYCAAWDDSLHVFGTGTKVTVLGQPKANPTVTLFPPS SEEL 527 3A64KC DIQMTQSPSSLSASVGDRVTITCRASQDITTYLAWLQQKPG KAPKSLIYSASTVQSGVPSRFSGSGSGTEFTLTISGLQPED FATYYCQQYNYYPITFGLGTRLEIKRTVAAPSVFIFPPSDE 528 3A650KC IILFLVATATGSWAQSALTQPRSVSGSLGQSVTISCTGSSS DVGRYNYVSWYQHHPGKAPKLMISDVNKRPSGVPDRFSGSK SGNTASLTISGLQAEDETDYYCCSYAGSYIWVFGG 529 3A67KC EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPG QAPRLLIYDASNRATGIPARFSGSGSDTDFTLTISSLEPED FAVYYCQQRGIWPLQITFGQGTRLEIKRTVAAPSVFIFPPS DE 530 3A779KC LSASVGDRVTITCRASQSIDRYLNWYQQKPGKAPKLLIYAA SSLHTDVPSRFSGSGAGTYFTLTITSLQPEDFATYYCQQSH SPSFGQESYSITFGQGTRLEIKRTVAAPSVFIFPPSD 531 3A816KC VTLSLSPGERATLSCRASQTISNNYLAWYQQKPGQAPRLLI YGASSGATGLPDRFSGSGSGTDFTLTISRLEPEDFAVYYCH QYALSPWTFGRGTKVEIKRTVAAPSVFIFPPSD 532 3A869KC 11L FLVATAT GVHS DIQMTQSPSSL SASVGDRVTIT GRAS Q 533 3A93LC QSVLTQPASVSGSPGQSITISCTGTNSDVGYSYVSWFQQHP GKVPKLLIYDVSRRSSGVSNRFSGSRSGNTASLTISGLRAE

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DEADYYCGSFTTSLTLVFGGGTKLAVLVSPS 534 For 42 bks EIVLTQSPGTLSLSPGERATLSCRASQSVSSRYLAWYQQKP GQAPRLIIYDASSRASGIPDRFSGSGSETDFTLTITRLEPE DFAVYYCQLYGTSPKFTFGQGTKLEIKRTVAAPSVFIFPPS D 535 Over515x DWMTQSPLSLPVTLGQPASISCRSSQSLVYSHGDTYLKCF QQRPGQSPRRPIYKVSNRDSGVPDRFSGSGSGTDFTLKISR VEAEDVGV 536 ZY29x GPATLSVSPGERATLSCRASQSLRNNLAWYQQKTGQSPRLL IYAVSTRATGIPPRFSGGGSGTEFTLTIDSLQSEDFAVYFC QQYDSPQWTFGQGTKVEIKRTVAAPSVFIFPPSD 537 ZY711S QSVLTQPASVSGSPGQSITISCTGTSNDVGGQNFVSWYQQH PGTAPQLLIYDVTNRPAGVSSRFSGSKSGNTASLTISGLRT EDEADYYCASFTILNGVDYVFGTGTKVTVLLSPSQPYL 538 Zb27x EIVLTQSPATLSVSPGERATLSCRAGQSVSSDLAWYQHKPG QAPRLLIYDASKRATGIPARFSGSGSGTDFTLTISSLEPED FAVYYCQHRTNWPPSIТ FGQGTRLEIKRTVAAPSVFIFPPS D 539 Zb41x EIVLTQSPGTLSLSPGERATLSCRASQSVSSNYLAWYQQKP GQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLSISRLEPE DFAVYYCQQYGTSSCTFGQGTKLEIKRTVAAPSVFIF 540 ZL45x EIVLTQSPGTLSLSPGDRAALSCRASETLSGNSLAWYQQKR GQPPRLLIFAASSRATGIPERFSGGGSGTDFTLTITRLEPE DFAVYFCQQYVDAPITFGQGTRLEIKRTVAAPSVFIFPPSD 541 Zb46x EIVLTQSPGTLSLSPGERATLSCRASQSVSSNNLAWYQQKP GQAPRLLMSGASSRATGIPDRFSGSGSGTDFTLTISRLEPE DFAVYHCQQYGSSPPTFGQGTKVEIKRTVAAPSVFIFPP 542 3b571c QSVLTQPRSVSGSPGQSVTISCTGTSSDVGGYNYVSWYQQH PGKAPKTMIFDVTKRPSGVPDRFSGSKSGNTASLTISGLQA E DEADYYC S S YAGRNT FYVFG T G T TVTVQVS P S Q R R R 543 Zb8x EIVLTQSPGTLSLSPGERATLSCRASQSVSSNLAWYAQKPG QAPRLIIYGASSRASAIPDRFRGSGSGTDFTLTISRLEPED FAVYYCQQYDDAPITFGHGTRLEIKRTVAAPSVFIFPPSDE 544 3BNC55KC DIQMTQSPSSLSASVGDKVTITCQTSAGYLNWYQQRRGRAP KLLMYDGSRLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT

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YYCQVYEFFGPGTRDLDLKSTVAA 545 3BNC60KC DIQMTQSPSSLSARVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPARFSGRRWGQEYNLTINNLQPEDVATY FCQVYEFIVPGTRRLDLKRTVAA 546 ZapsZks DIQMTQSPSSVSASVGDRVTITCQASRDTDNSLTWYQQKPG RPPKLLIYHWNLGPGVPSRFSGSASSATQSTLIISDFQPD DVATYFCQNYEFFGPGTKVEIKRTVAAPSVFIFPPSDEQ 547 ZYObx DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFWPGTRDLLKRTVAAPSVFIFPPSD 548 PS bks DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFWPGTRDLLKRTVAAPSVFI FPPSD 549 PS 8 Oke DIQMTQSPSSLSARVGDTVTFTCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRGWGQEYNLTINNLQPEDIATY FCQVYEFAVPGTRDLDLKRTVAAPSVFIFPPSD 550 ZY83x DIQMTQSPSSLSARVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLETGVPSRFTGRRWGQEYNLTINNLQPEDIATY FCQVYEFIVPGTRRLDLKRTVAAPSVFIFPPSD 551 ZY91x DIQMTHSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLETGVPSRFTGRRWGQEYNLTINNLQPEDIATY FCQVYEFIVPGTRLDLKRTVAAPSVFIFPSD 552 Zb21x DIQMTQSPSSLSARVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLETGVPSRFTGRRWGQEYNLTINNLQPEDIATY FCQVYEFIVPGTRRLDLKRTVAAPSVFIFPPSD 553 Zps102x DIQMTQSPSSLSASVGDRVTITCQASQGISNSLNWYQQKPG KAPRLLIYGTSTLQRGVPSRFSGSGSGTRFTVTINSLQPED IATYFCQHNEFFGRGTKVDIKRTVAAPSVFIFPPSDEQL 554 Zps104x DIQMTQSPSSLSASIGDRVNITCQASRDTGSALNWYQQKVG RPPRRLLISAAVSNLGAGVPSRFSGRRSGTQSTLTINTLQPED IATYFCQHYEFFGPGTKVDIKRTVAAPSVFIFPPSDEQ 555 3bncl05ks DIQMTQSPSSLSASVGDTVTFTCQANGYLNWYQQRRGKAPK LLIYDGSRLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFAVPGTRLDLKRTVAAPSVFIFPSD 556 Zps107x DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP

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KLLMYDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRRLDLKRTVAAPSVFIFPPSD 557 3bncl08kc DIQMTQSPSSLSARVGDKVTITYQTSAGYLNWYQQRRGRAP KLLMYDGSRLVTGAPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRLDLKRTVAAPSVFIFPPSD 558 3bncll7kc DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFWPGTRDLLKRTVAAPSVFIFPPSD 559 3bncl34kc DIQMTQSPSSLSASVGDTVTINCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 560 3bncl42kc DIQMTQSPSSLSASVGDTVTITCHTNKGYLNWYQQRRGRAP KLLMFDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEVFGPGTRLDLKRTVAAPSVFIFPSD 561 3bncl51kc DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 562 3bncl53kc DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRLSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 563 3bncl56kc DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQKRGRAP KLLMYDGSKLVTGVPSRLSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 564 3bncl58kc DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRLSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 565 3bncl59kc DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 566 3bncl5kc DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRLSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 567 3bncl7 6kc DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQAEDIATY FCQVYEFAVPGTRDLLKRTVAAPSVFIFPPSD

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568 3bncl93kc DIQMTQSPSSLSARVGDKVTITCQTSAGYLNWYQQRRGRAP KLLMYDGSRLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 569 3bncl96kc DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPK LLMYDGSTLERGVPARFSGRRWGQEYNLTINNLQPEDVATY FCQVYEFIVPGTRRLDLKRTVAAPSVFIFPSD 570 3bnc31kc DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP KLLMCDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRLDLKRTVAAPSVFIFPPSD 571 3bnc42kc DIQMTQSPSSLSASVGDTVTITCQTTKGYLNWYQQRRGRAP KLLMFDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDLAT YYCQVYEFFGPGTRLDLKRTVAAPSVFIFPPSD 572 3bnc53kc DIQMTQSPSSLSASVGDTVTITCHTNKGYLNWYQQRRGRAP KLLMFDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEVFGPGTRLDLKRTVAAPSVFIFPSD 573 3bnc62kc DIQMTQSPSSLSARVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLETGVPSRFTGRRWGQEYNLTINNLQPEDIATY FCQVYEFIVPGTRRLDLKRTVAAPSVFIFPPSD 574 3bnc65kc DIQMTQSPSSLSARVGDTVTFTCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFAVPGTRDLDLKRTVAAPSVFIFPSD 575 3psbbx DIQMTQSPSSLSASVGDTVTITCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRLSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 576 3bnc75kc DIQMTQSPSSLSARVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFWPGTRDLLKRTVAAPSVFI FPPSD 577 3bnc7 9kc DIQMTQSPSSLSARVGDTVTFTCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFAVPGTRDLDLKRTVAAPSVFIFPSD 578 3bnc81kc DIQMTQSPSSLSASVGDTVTINCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRLDLKRTVAAPSD 579 3bnc84kc DIQMTQSPSSLSASVGDTVTINCQTNKGYLNWYQQRRGRAP KLLMYDGSKLVTGVPSRFSGRRWGTQYNLTIGSLQPEDIAT YYCQVYEFFGPGTRDLDLKRTVAAPSVFIFPPSD 580 3bnc87kc DIQMTQSPSSLSARVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFWPGTRDLLKRTVAAPSVFI FPPSD 581 3bnc89kc DIQMTQSPSSLSASVGDKVTITCQTSAGYLNWYQQRRGRAP KLLMYDGSRLVTGVPSRFSGRRWGTQYNLTIGSLQPEDVAT YYCQVYEFFGPGTRDLLKRTVAAPSVFIFPPSD 582 3bnc91kc DIQMTQSPSSLSARVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFAVPGTRLDLKRTVAAPSVFIFPSD 583 3bnc95kc DIQMTQSPSSLSASVGDTVTITCQANGYLNWYQQRRGKAPK LLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATY FCQVYEFIVPGTRRLDLKRTVAAPSVFIFPSD

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Table 1

Forward leader sequence primers VH1 LEADER-A ATGGACTGGACCTGGAGGAT SEQ ID NO:591 VH1 LEADER-B ATGGACTGGACCTGGAGCAT SEQ ID NO:592 VH1 LEADER-C ATGGACTGGACCTGGACAAT SEQ ID NO:593 VH1 LEADER-D GGCCTTCTCTTTGTGGTGGC SEQ ID NO:594 VH1 LEADER-E ATGGACTGGACCTGGAGGGT SEQ ID NO:595 VH1 LEADER-F ATGGACTGGATTTGGAGGAT SEQ ID NO:596 VH1 LEADER-G AGGTTCCTCTTTGTGGTGGCAG SEQ ID NO:597 VH3 LEADER-A TAAAAGGTGTCCAGTGT SEQ ID NO:598 VH3 LEADER-B TAAGAGGTGTCCAGTGT SEQ ID NO:599 VH3 LEADER-C TAGAAGGTGTCCAGTGT SEQ ID NO:600 VH3 LEADER-D GCTATTTTTAAAGGTGTCCAGTGT SEQ ID NO:601 VH3 LEADER-E TAGAAGGTGTCCAGTGT SEQ ID NO:602 VH3 LEADER-F TTAAAGCTGTCCAGTGT SEQ ID NO:603 VH4 LEADER-A ATGAAACACCTGTGGTTCTTCC SEQ ID NO:604 VH4 LEADER-B ATGAAACACCTGTTTCTT SEQ ID NO:605 VH4 LEADER-C ATGAAGCACCTGTGGTTCTT SEQ ID NO:606 VH4 LEADER-D ATGAAACATCTGTGGTTCTT SEQ ID NO:607 VH5 LEADER-A TTCTCCAAGGAGTCTGT SEQ ID NO:608 VH5 LEADER-B CCTCCACAGTGAGAGTCTG SEQ ID NO:609 VH6 LEADER-A ATGTCTGTCTCCTTCCTCATC SEQ ID NO:610 VH7 LEADER-A GGCAGCAGCAACAGGTGCCCA SEQ ID NO:611 Reverse constant region primers 3' Cg CHI (g amma) GGAAGGTGTGCACGCCGCTGGTC SEQ ID NO:612 3' IgG (internal) GTTCGGGGAAGTAGTCCTTGAC SEQ ID NO:613

table 2

Floor Treasure Year of birth Year of diagnosis CD4+ T cells/µl Virus copies/ml Clinical status ptl Male IN 1948 1985 354 4722 nonprogressor pt3 Male IN 1965 2002 427 880 nonprogressor pt8 Male IN 1962 1989 580 <50 selective controller pt!2 Male No data No data No data No data No data No data

Table FOR

Name AT vh D JH (-) CDR3 (a.k.) SEQ ID NO 3BNC4 1-2 7-27 2/6 3 RHSDYCDFDV 614 3BNC23 1-2 6-25/3-3 2/6 3 QRSDFWDFDV 615 3BNC42 1-2 7-27 2/6 3 RHSDYCDFDV 616 3BNC53 1-2 3-3 2/6 3 RHSDYCDFDV 617 3BNC55 1-2 3-3/6-19/5-12 2/6 3 RHSDYCDFDI 618 3BNC62 1-2 6-25/6-13/6-6 2/6 3 QRSDYWDFDV 619

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3BNC65 1-2 6-25/6-6 2/6 3 QRSD YWDFD V 620 3BNC66 1-2 7-27 2/6 3 RHTDYCDFDV 621 3BNC72 1-2 7-27 2/6 3 RHSDYCDFDV 622 3BNC79 1-2 6-25/6-6 2/6 3 QRSD YWDFD V 623 3BNC81 1-2 7-27 2/6 3 RHSDYCDFDV 624 3BNC89 1-2 3-3/6-19/5-12 2/6 3 RHSDYCDFDI 625 3BNC91 1-2 2-21/6-25 2/6 3 RRSDYCDFDV 626 3BNC95 1-2 6-25/2-8 2/6 3 QRSD YWDFD V 627 3BNC105 1-2 6-6/6-25 2/6 3 QRSD YWDFD V 628 3BNC107 1-2 7-27/3-3 2/6 3 RHSDYCDFDV 629 3BNC108 1-2 3-3/6-19/6-25 2/6 3 RHSDYCDFDI 630 3BNC117 1-2 6-25/2-8 2/6 3 QRSD YWDFD V 631 3BNC134 1-2 7-27 2/6 3 RHSDYCDFDV 632 3BNC142 1-2 3-3 2/6 3 RHSDYCDFDV 633 3BNC151 1-2 7-27/4-17/3-3 2/6 3 RHSDYCDLDV 634 3BNC156 1-2 3-3/7-27 2/6 3 RHSDYCDFDV 635 3BNC159 1-2 7-27 2/6 3 RHSDYCDFDV 636 3BNC176 1-2 6-25/6-6 2/6 3 QRSD YWDFD V 637 3BNC196 1-2 6-25/6-6/6-13 2/6 3 QRSD YWDFD V 638 3BNC6 1-2 3-16/1-7 2 1 PLRGGDTWHYH S 639 3BNC101 1-2 1-7/3-16 2 1 PLRGGDTWHYH S 640 3BNC102 1-2 3-22/1-26/1-20 2 3 PHSPDDAWSLD V 641 3BNC126 1-2 3-22/1-26/1-20 2 3 PHSPDDAWSLD V 642 3BNC149 1-2 3-22/1-26/1-20 2 3 PHSPDDAWSLD V 643 3ANC3 1-2 2-21/2-15 1/2 1 PRGGRDNW SFH V 644 3ANC42 1-2 ND 2 2 PKSGRDYWSFD L 645 3BNC3 1-69 5-5/5-18/5-24 3 2 ATGYSYGYLDA 646

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FDI 3BNC8 1-24 5-24/4-17 4 3 EPREMGTLT AG FEY 647 3BNC48 1-69 3-3 4 5 GQTDLNDDLWS DYSTPGFDY 648 3ANC38 1-69 3-3 4 5 GQTDLNDDFWS EYSTPGFDY 649 3BNC49 1-69 3-22/6-19/5-12 6 3 GEFDSSGFDYE SWYPYYMDV 650 3BNC58 1-24 3-16/3-10 4/5 2 APRLELGEL S SG FH Y 651 3BNC78 1-24 4/5 2 APRLDLGELSS GFHF 652 3BNC78 1-24 4/5 2 APRLDLGEL S S GFHF 653 3BNC71 1-24 1-24 4/5 3 DNPLLQ S GEF S S SLDN 654 3BNC71 1-24 1-24 4/5 3 DNPLLQ S GEF S S SLEN 655 3BNC144 1-69 3-9/5-5 4 3 AQ GDILTEGYF DY 656

Name AT (+) Length TC mutations Primer set k/1 Vk/1 Jk/1 (-) 3BNC4 1 10 72 new To 1D-33 3 1 3BNC23 1 10 79 new To 1D-33 3 1 3BNC42 2 10 69 new To 1D-33 3 1 3BNC53 2 10 74 new To 1D-33 3 1 3BNC55 2 10 64 new To 1D-33 1/3 1 3BNC62 1 10 81 new To 1D-33 3 1 3BNC65 1 10 82 new To 1D-33 3 1 3BNC66 2 10 69 new To 1D-33 3 1 3BNC72 1 10 72 new To 1D-33 3 1

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3BNC79 1 10 76 new To 1D-33 3 1 3BNC81 2 10 71 new To 1D-33 3 1 3BNC89 2 10 68 new To 1D-33 3 1 3BNC91 2 10 76 new To 1D-33 3 1 3BNC95 1 10 72 new To 1D-33 3 1 3BNC105 1 10 77 new To 1D-33 3 1 3BNC107 2 10 69 new 3BNC108 2 10 62 new To 1D-33 3 1 3BNC117 1 10 72 new To 1D-33 3 1 3BNC134 2 10 71 new To 1D-33 3 1 3BNC142 2 10 72 new To 1D-33 3 1 3BNC151 2 10 69 new To 1D-33 3 1 3BNC156 2 10 72 new To 1D-33 3 1 3BNC159 2 10 71 new To 1D-33 3 1 3BNC176 1 10 72 new To 1D-33 3 1 3BNC196 1 10 78 new To 1D-33 3 1 3BNC6 3 12 55 new To 1D-33 1/3 1 3BNC101 3 12 54 new 3BNC102 1 12 63 new To 1D-33 1/3 1 3BNC126 1 12 65 new 3BNC149 1 2 68 new 3ANC3 3 12 59 new To 1D-33 3 1 3ANC42 2 12 53 new To 1D-33 3 1 3BNC3 0 14 22 new L 1-44 1 2 3BNC8 1 14 21 old To 3-11 2 0 3BNC48 0 20 18 new 3ANC38 0 20 12 new 1 1-47 1/6 2 3BNC49 0 20 23 old To 3-20 3 3BNC58 1 15 16 old To 3-11 2 0 3BNC78 2 15 38 old 3BNC78 2 15 39 old 3BNC71 0 16 22 old To 3-11 5 3BNC71 0 16 17 old To 3-11 5 3BNC144 0 13 15 old k/1 1-44/1-47 1 2

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data 3BNC6 QHYEF 681 1 5 44 + 24 3BNC101 No data No data 1 3BNC102 QHYEF 682 1 5 34 - 1 3BNC126 No data No data 1 3BNC149 No data No data 1 3ANC3 QHYEF 683 0 5 47 + 1 3ANC42 QQYEF 684 1 5 41 No data 4 3BNC3 AAWDDT LYV 685 0 9 19 CD4i + 7 3BNC8 QHRSIWP LMCT 686 2 eleven 10 CD4i + 3 3BNC48 No data No data 3ANC38 GAWDDT LYV 687 0 9 8 CD4i - 2 3BNC49 No data CD4i No data 2 3BNC58 QQRTIWP PGCS 686 1 eleven 10 CD4i No data 2 3BNC78 No data No data 1 3BNC78 No data No data 2 3BNC71 No data CD4i No data 1 3BNC71 CD4i No data 1 3BNC144 No data 1 9 CD4i No data 1

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Table 3B

Name vh D JH (-) CDR3 (a.k.) SEQ ID At NO 1NC2 1-46 3-22/5-5 4/5 4 NEADYHDGNGHHSLR GMFD Y 687 1NC3 1-46 6-19 4/5 3 AEAESQSHSRPIMFDF 688 1NC7 1-46 6-19/1- 14 4/5 3 AEAESQSHSRPIMFDS 689 1NC9 1-46 5-12/2-8 4/5 4 QDSDFHDGHGHTLRG MFD S 690 INC 18 1-46 1-14/2- 21 4/5 2 NEPQYHSLPGMFDY 691 1NC24 1-46 3-16 4/5 3 NEPQYHDGNGHSLPG MFD Y 692 1NC29 1-46 3-16/6- 19 4/5 3 NEPQYYDGSGHSLPG MFD Y 693 1NC33 1-46 5-12 4/5 5 LEADGDDYSPKMVD Y 694 1NC46 1-46 3-9/3-16 4/5 3 READYHDGNGHTLPG MFDF 695 1NC48 1-46 3-9/6-19 4/5 2 NEPQYFDGSGHSLPG MFD Y 696 1NC52 1-46 3-16/6- 19 4/5 3 NEPQYYDGSGHSLPG MFD Y 697 1NC56 1-46 5-12/3-9 4/5 5 LEADGDDYSPKMFDH 698 1NC60 1-46 3-22/1- 26 1/5 4 LEAESDSHSRPIMFD H 699 1NC66 1-46 3-16 4/5 2 NEPQYHDGNGHSLPG MFDF 700 1NC70 1-46 3-16/6- 19 4/5 3 NEPQYYDGSGHSLPG MFD Y 701 1NC72 1-46 6-19/1- 14 4/5 3 AEAESQSHSRPIMFDF 702 1NC94 1-46 6-13/6- 19 4/5 3 AEAASD SHSRPIMFD H 703 1NC95 1-46 3-16/6- 4/5 4 LEADGSDYSPKMFDF 704

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19 I.N.C. 107 1-46 3-3/5-12 4/5 5 LEADGDDYSPKMFDY 705 I.N.C. 108 1-46 3-9/3-16 4/5 4 READYHDGNGHTLPG MFDF 706 I.N.C. 109 1-46 5-1/6-19 4/5 5 LEADGDDYSPKMFDY 707 1NC110 1-46 5-24/6- 19 4/5 4 LEADGDNYSPKMVD Y 708 1NC116 1-46 2-21 4 2 NEPQYHSLPGMFDY 709 1NC118 1-46 3-9/5-12 4 3 LEADGGDYSPKMFDY 710 I.N.C. 122 1-46 3-16/3-3 4 4 LEADGADYSPKMFDF 711 1NC123 1-46 6-19 4 3 AEAESQ SHSRPIMFD 712 1NC127 1-46 6-13/6- 19 4/5 3 AEAASDSHSRPIMFD H 713 1B344 1-46 3-22/1- 26 1/5 4 LEAESDSHSRPIMFD H 714 1B2416 1-46 1-14/3- 16 4 4 NEPQYHDDNGHSLPG MID-Y 715 1B2503 1-46 6-19 5 3 AEAESQSHSRPIMFDS 716 1B2573 1-46 3-22 4/5 2 NEPQYHDGNGHSLPG MFD S 717 1NC5 1-69 3-3 3 1 GRQTFRAIWSGPPVV FDI 718 1NC126 1-69 3-3 3 1 GRQTFRAIWSGPPAV FDI 719 I.N.C. 16 4-34 3-10 5 2 AVAGLWFEDAYNWF GP 720 1NC21 4-34 3-10 5 2 AVKGLWFDETYTWF GP 721 1NC54 4-34 3-10 5 2 AVKGFWFDEPSTWFG P 722 1NC57 4-34 3-10 5 2 AVKGFWFDDPYTWF GP 723 1NC115 4-34 3-10 5 2 AVKGFWFDEVYNWF 724 GP

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Name AT (+) Length TC mutations Primer set k/1 Vk/1 Jk/1 ABOUT 1NC2 2 19 74 new 1 1-47 3 1 1NC3 2 16 86 new 1 1-47 6/7 1 1NC7 2 16 77 new 1 1-47 6/7 1 1NC9 4 19 67 new 1 1-47 3 1 INC 18 1 14 85 new 1NC24 2 19 79 new 1 1-47 3 1 1NC29 1 19 87 new 1NC33 0 15 84 new 1 1-47 3 2 1NC46 3 19 85 new 1 1-47 3 1 1NC48 1 19 88 new 1 1-47 3 1 1NC52 1 19 82 new 1 1-47 3 1 1NC56 2 15 91 new 1 1-47 3 1 1NC60 3 16 72 new 1 1-47 3 1 1NC66 2 19 91 new 1 1-47 3 1 1NC70 1 19 85 new 1 1-47 3 1 1NC72 2 16 77 new 1 1-47 6/7 1 1NC94 3 16 81 new 1 1-47 3 2 1NC95 0 15 93 new I.N.C. 107 1 15 90 new 1 1-47 3 1 I.N.C. 108 3 19 85 new 1 1-47 3 1 I.N.C. 109 1 15 85 new 1NC110 1 15 88 new 1NC116 1 14 83 new 1NC118 0 15 86 new 1 1-47 3 1 I.N.C. 122 1 15 94 new 1 1-47 3 1 1NC123 2 16 78 new 1 1-47 3 1 1NC127 3 16 81 new 1 1-47 3 2 1B344 3 16 72 new 1 1-47 3 1 1B2416 2 19 81 new 1B2503 1 16 78 new 1 1-47 3 1 1B2573 2 19 81 new 1NC5 2 18 47 new To 3-11 2 0 1NC126 2 18 47 new I.N.C. 16 0 16 75 new To 1D-39 2/3 0 1NC21 1 16 58 new 1NC54 1 16 59 new 1NC57 1 16 61 new 1NC115 1 16 58 new

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AT name CDR3 (a.k.) SEQID NO (+) Length LC mutations Binding NEUT # relatives 1NC2 AVYDSS LSLGL 725 0 eleven 47 + 15 1NC3 ATYD SQ RSIRL 726 2 eleven 55 + 1 1NC7 ATYD SQ G STRL 727 1 eleven 51 + 1 1NC9 AAYDST FSLPV 728 0 eleven 53 ? + 2 1NC18 No data No data 1 1NC24 AAYDSS LSLRL 729 0 eleven thirty + 2 1NC29 No data No data 1 1NC33 ATYDTD LSLRL 730 1 eleven 49 + 1 1NC46 AAYDSA VSLP V 731 0 eleven 52 No data 1 1NC48 AAYDST LSLRL 732 1 eleven 37 No data 1 1NC52 AAYDST FSLPV 733 0 eleven 54 No data 1 1NC56 ATYDTG LSLRL 734 1 eleven 58 No data 1 1NC60 ATYD SG WSIRL 735 1 eleven 46 + 3 1NC66 AAYDST LSLRL 736 1 eleven 33 No data 1

- 97 042696

1NC70 AAYDST LSLRL 737 1 eleven 40 No data 1 1NC72 ATYD SQ GSTRL 738 1 eleven 51 + 2 1NC94 ATYD SD GSIRL 739 1 eleven 41 - 5 1NC95 No data No data 1 1NC107 ATYDTG LSLRL 740 1 eleven 58 No data 1 1NC108 AAFD SA LSLPL 741 0 eleven 51 + 1 1NC109 No data No data 1 1NC110 No data No data 1 1NC116 No data No data 1 1NC118 ATYDTG LSLRL 742 1 eleven 54 No data 1 1NC122 GTYDTS LSLRL 743 1 eleven 57 No data 1 1NC123 ATYD SH GSIRL 744 2 eleven 48 - 1 1NC127 ATYD SD GSIRL 745 1 eleven 41 ? + 5 1B344 ATYD SG WSIRL 746 1 eleven 46 + 1 1B2416 No data No data 1 1B2503 GTYD SQ GSTRL 743 1 eleven 49 No data 1 1B2573 No data - 2 1NC5 QHRSNW PWT 744 2 9 CD4BS + 1 1NC126 No data No data 1 1NC16 QQ SF AV PYT 745 0 9 35 No data No data 1 1NC21 No data No data No data 1 1NC54 No data No data No data 1 1NC57 No data No data No data 1 1NC115 No data No data No data 1

- 98 042696

Table 3C

Name AT vh D JH (-) CDR3 (a.k.) SEQ ID NO 8ANC13 1-46 3-16 6 4 DGLGEVAPDYRYGID V 746 8ANC22 1-46 3-16 6 3 DGLGEVAPAYLYGID A 747 8ANC26 1-46 3-16 6 3 DGLGEVAPAYLYGID A 748 8ANC37 1-46 3-16 6 3 DGLGEVAPAYLYGID A 749 8ANC41 1-46 3-16 6 3 DGLGELAPAYHYGID V 750 8ANC50 1-46 3-16 6 3 DGLGELAPAYQYGID V 751 8ANC88 1-46 3-16 6 4 DGLGEVAPDYRYGID V 752 8ANC127 1-46 3-16 6 3 DGLGEVAPAYLYGID A 753 8ANC131 1-46 3-16 6 3 DGLGEVAPDYRYGID V 754 8ANC142 1-69 3-3 No data 2 TSTYDQWSGLHHDG VMAF S S 755 8ANC46 1-69 3-22/2- 15 3 2 S SGNFEFAFEI 756 8ANC191 1-69 3-22/2- 15 3 2 S S GNYDFAYDI 757 8ANC196 1-69 3-22/2- 15 3 2 S S GNYDFAFDI 758 8ANC14 1-24 6-13/5- 5 4 4 ADRFKVAQDEGLFVI F.D.Y 759

- 99 042696

8ANC34 1-24 6-13/5- 5 4 4 ADPFKVAQDEGLYVI F.D.Y 760 8ANC58 1-24 6-13/5- 5 4 4 ADPFKVAQDEGLYVI F.D.Y 761 8ANC168 1-24 6-13/5- 5 4 4 ADPFKVAQDEGLFVI F.D.Y 762 8ANC5 1-69 4-17/3- 10 6 8 DRGDTRLLDYGDYED ERYYYGMDV 763 8ANC7 1-69 4-17/3- 10 6 8 DRGDTRLLDYGDYED ERYYYGMDV 764 8ANC9 1-69 4-17/3- 10 6 8 DRGDTRLLDYGDYED ERYYYGMDV 765 8ANC77 1-69 4-17/3- 10 6 8 DRGDTRLLDYGDYED ERYYYGMDV 766 8ANC107 1-69 4-17/3- 10 6 8 DRGDTRLLDYGDYED ERYYYGMDV 767 8ANC108 1-69 4-17/3- 10 6 8 DRGDTRLLDYGDYED ERYYYGMDV 768 8ANC137 1-69 4-17/3- 10 6 8 DRGDTRLLDYGDYED ERYYYGMDV 769 8ANC16 1-69 2-2 3 2 DRS AIGYC S SISC YK GSFDI 770 8ANC24 1-24 2-2 6 1 GGLYCSSISCIMDV 771 8ANC25 1-24 2-2 6 1 GGLYCSSISCIMDV 772 8ANC38 3-43 3-16 5 1 NGFD V 773 Name AT (+) Length TC mutations Primer set k/1 Vk/1 Jk/1 (-) 8ANC13 1 16 75 new To 3-11 2/3 1 8ANC22 0 16 85 new 8ANC26 0 16 76 new To 3-11 2/3 1 8ANC37 0 16 82 new To 3-11 2/3 1 8ANC41 1 16 71 new To 3-11 2/3 1

- 100 042696

8ANC50 0 16 71 new To 3-11 2/3 1 8ANC88 0 16 73 new To 3-11 2/3 1 8ANC127 0 16 86 new 8ANC131 1 16 75 new To 3-11 2/3 1 8ANC142 2 20 72 new To 1-5 1/5 1 8ANC46 0 And thirty old 1 1-40 3 1 8ANC191 0 And 28 old 8ANC196 0 And 25 old 8ANC14 1 18 AND old To 3-11 4 0 8ANC34 0 18 10 new 8ANC58 0 18 18 new 8ANC168 1 18 AND new 8ANC5 3 24 40 old To 1D-33 2 0 8ANC7 3 24 37 new 8ANC9 3 24 35 old 8ANC77 3 24 50 old 8ANC107 3 24 38 old 8ANC108 3 24 37 old 8ANC137 3 24 37 new 8ANC16 1 21 12 old To 3-15 2 0 8ANC24 0 14 12 old To 3-15 1 0 8ANC25 0 14 6 old 8ANC38 0 5 70 new 1 2-11 3 0

- 101 042696

8ANC41 QEYS STP YN 777 0 9 42 + 2 8ANC50 QEYS STP YN 778 0 9 46 CD4BS + 2 8ANC88 QEYS STP YN 779 0 9 46 No data 1 8ANC127 No data No data 1 8ANC131 QEYS STP YN 780 0 9 45 CD4BS + 1 8ANC142 QQYDTYP GT 781 0 9 43 ? + 2 8ANC46 QSYDRSL RGS V 782 1 eleven thirty No data No data 1 8ANC191 No data No data 1 8ANC196 No data No data 1 8ANC14 QQRANWR LLT 783 2 10 9 CD4i + 2 8ANC34 No data No data 5 8ANC58 No data No data 3 8ANC168 No data No data 1 8ANC5 QQYSNLP YT 784 0 9 17 CD4i - 2 8ANC7 No data No data 2 8ANC9 No data No data 1 8ANC77 No data No data 3 8ANC107 No data No data 2 8ANC108 No data No data 4 8ANC137 No data No data 1 8ANC16 QQYYQWL 785 0 10 13 No data No 8 S YT data 8ANC24 QQYNHWP QT 786 0 9 7 CD4i + 1 8ANC25 No data No data 1 8ANC38 CLKKTS S YV 787 2 9 41 CORE + 2

- 102 042696

Table 3D

Name At vh D JH (-) CDR3 (a.k.) SEQ ID NO 12A1 1-2 5-12/3-10 4/5 4 DESGDDLKWHL HP 787 12A2 1-2 4-17 4/5 3 DGSGDDTSWHLH R 788 12A4 1-2 5-12/3-10 4/5 4 DESGDDLKWHL HP 789 12 Ab 1-2 1-26/3-10 4/5 2 DGSGDATSWHLH R 790 12A7 1-2 1-26 4/5 4 DGSGDARDWHL D.P. 791 12A9 1-2 3-3 4/5 5 DRRDDDRAWLL D.P. 792 12А12 1-2 1-26/3-10 4/5 4 DGSGDDTSWHLD P 793 12A13 1-2 1-26 4/5 4 DGSGDDTSWYLD P 794 12A20 1-2 1-26 4/5 3 DGSGDARDWHL HP 795 12A22 1-2 3-16 4/5 4 DGGGDDRTWLL DA 796 12A23 1-2 3-3 4/5 5 DRRDDGLDWLL D.P. 797 12A27 1-2 1-26/3-10 4/5 3 DGSGDDTSWHLH P 798 12A46 1-2 3-10 4/5 1 GGGDGRNWHLH P 799 12А55 1-2 1-26 4/5 4 DGSGDDRNWHL D.P. 800 12A56 1-2 1-26 4/5 4 DESGYDLNWHL D S 801 Name At (+) Length # TC mutations Primer set k/1 Vk/1 Jk/1 (-) 12A1 2 13 60 new k ID-33 3 0 12A2 2 13 67 new k ID-33 3 10 12A4 2 13 59 new k ID-33 3 0 12 Ab 2 13 61 new k ID-33 3 1 12A7 1 13 62 new k ID-33 3 1 12A9 3 13 62 new k ID-33 3 1 12А12 1 13 60 new k ID-33 3 1 12A13 0 13 61 new k ID-33 3 1 12A20 3 13 61 new k ID-33 3 1 12A22 1 13 61 new k ID-33 3 1 12A23 2 13 51 new k ID-33 3 1 12A27 2 13 68 new k ID-33 3 1 12A46 3 13 62 new k ID-33 3 1 12А55 1 13 63 new k ID-33 3 2 12A56 1 13 66 new k ID-33 3 1

- 103 042696

W 12 Ab A VLEF 804 0 5 39 + 1 12A7 A VLEF 805 0 5 41 No data 2 12A9 QLFEF 806 0 5 39 No data 1 12А12 A VLEF 807 0 5 41 CD4BS + 1 12А13 AVVEF 808 0 5 41 No data 1 12А20 AALEF 809 0 5 40 + 1 12A22 S VYEF 810 0 5 39 + 2 12A23 QLFEF 811 0 5 39 + 1 12A27 A VLEF 812 0 5 40 No data 1 12A46 ASLEF 813 0 5 43 + 1 12А55 EVYEF 814 0 5 37 + 1 12A56 ESFQW 815 0 5 37 No data 1

Table 3E

Name At vh D JH ABOUT CDR3 (a.k.) SEQ ID NO ZV191 1-2 6-25/6-13/6-6 2/6 3 QRSDYWDFDV 816 SW6 4-39 3-9/3-10 3 2 IP YHSES YYK VVI GGFD V 817 SG8 1-69 4-17/3-22 4 3 DHGDPRTGYYFD Y 818 ZV27 3-64 3-9/1-26/4-17 5 1 GPLLRYLD S 819 ZV41 1-24 3-16 6 4 KAKDYYYES SD Y SPYYYYYMDV 820 ZV46 4-31 3-3/2-8 4/5 0 GSGRWTIGARIY FDN 821 SW 144 3-30 3-3/3-10/3-16 4/5 2 TPPHYDVLTGYP S SVLEF 822 ZV117 1-69 5-5/5-18/5-24 3 2 ATGYSYGYLDAF DI 823 ZA869 4-4/4-59 6-19/5-12/1- 26 4 2 EKGQWLTVPPYY FD S 824 ZA228 5-51 3-3/2-2 6 1 TRCF GANCFNFM D V 825 ZA461 1-46 2-2 4 1 PEP S SIVAPLYY 826 FOR18 1-69 3-10/5-24 3 3 DPQVEVRGNAFD I 827 FOR125 1-46 1-20/1-7/3-10 3 2 PQYNLGRDPLDV 828 ZA255 4-59 3-3/3-9 4 3 ADYDLLT S SYHF D S 829 3A233 4-59/4-61 3-3/4-17 4/5 3 LDGEAFRYYLDL 830

- 104 042696

Name At (+) Length # TC mutations Primer set k/1 Vk/1 Jk/1 (-) ZV191 1 10 81 new To 1D-33 3 1 SW6 1 18 50 new To 1-9 1/3 0 SG8 2 13 50 new To 3-20 1/5 2 ZV27 0 9 18 old To 3-11 1/5 0 ZV41 2 22 17 old To 3-20 2 0 ZV46 2 15 22 old To 3-20 1/4 0 ZV144 1 18 23 old To 3-15 1/5 0 ZV117 0 14 22 new 1 1-44 1 2 ZA869 1 1 33 old To 1D-39 5 0 ZA228 1 1 34 old To 4-1 3 0 ZA461 0 1 15 old To 3-20 1 0 FOR18 1 1 40 old To 1D-39 5 0 FOR125 1 1 22 old To 3-20 1 0 ZA255 1 1 35 old 1 7-43 3 0 3A233 1 1 32 old 1 2-14 2/3 0

Name At CDR3 (a.k.) SEQ ID NO (+) Length Binding NEUT # kinship nnikov ZV191 QVYEF 831 0 5 CD4BS + 7 SW6 QQLAT 832 0 5 GP41 + eleven SG8 QQYDD APIT 833 0 9 GP41 - 9 ZV27 QHRTNWPPSIT 834 2 eleven CD4i - 3 ZV41 QQYGTSSCCT 835 0 9 CD4i - 2 ZV46 QQYGSSPPT 836 0 9 GP41 No data 2 SW 144 QQYNNWPPIT 837 0 10 No data No data 4 ZV117 AAWDDTLYV 838 0 9 No data No data 1 ZA869 QQSHSPS 839 1 7 CD4BS + 1 ZA228 QQYYISP 840 0 7 VAR + 4 ZA461 QQYGTLHPRT 841 2 10 GP41 - 3 FOR18 QQTYTSPIT 842 0 9 GP41 - 2 FOR125 QQYGLSPWT 843 0 9 GP41 - 4 ZA255 LLLPYYGGPWI 844 0 eleven GP41 - 2 3A233 S SFTPTNTLV 845 0 10 GP41 - 2

Table 3F

Name At vh D JH (-) CDR3 (a.k.) SEQID NO 1B2434 15341 3-22/5-5 1 4 NEADYHDGNGHHSLRGM F.D.Y 846 1B218 1-69 3-3 3 1 GRQTFRAIWSGPPVVF DI 847 1B331 4-34 3-9/3-3 6 3 RYFDWSPFRRDTYGTD V 848 1B2174 4-34 3-9/3-3 6 3 RYLDWSPIGRDTYGTD V 849 1B2055 1-69 2-21 2/5 1 GLCRGGNCRLGPSGWL D.P. 850 1B2133 1-3 4-17/2- 21 4 1 VAYVHVVTTRSLDN 851 1A64 4-59 5-5/5-18 6 2 HEAPRYSYAFRRYYHY 852

- 105 042696

GLD V 1A621 4-59 3-3/3-9 6 1 VISGRITIFYYNYIDV 853 1A577 3-48 3-10/3- 16 1 3 GTLWFGESGLRDDH 854 1A732 3-7/3- 73 3-22/3- 10 6 2 NRRVAMPEAMILSFYM D V 855 1A74 4-34 3-3/3-9 4 1 VVPMFSIFGVVKANYF DY 856 1A695 4-59 3-3/3-9 3 2 AGLDYNFWNGKGRKG AFD V 857 1A479 1-69 3-22 4 1 GFRGSPFSSGSLYFDS 858 1A182 1-69 4-17/1- 26 6 6 AVITDLHTFGDYELEDP SYYYMDV 859 1A693 3-23 7-27/3- 22 4 1 RGRRQIGDY 860 1A79 5-51 3-9/3-3 3 4 SYYDFSIGDGNDAFDV 861 1A27 3-11 3-6/5-5 5 2 DTTTFTTFGGGPNMGG FDP 862

Name At (+) Length # TC mutations Primer set k/1 Vk/1 Jk/1 (-) 1B2434 2 19 74 new 1 1-47 3 1 1B218 2 18 47 new To 3-11 2 0 1B331 3 17 40 new To 4-1 1/4 0 1B2174 2 17 41 new To 4-1 1/4 0 1B2055 2 18 62 new To 3-15 1 2 1B2133 1 14 22 new To 1D-39 1 0 1A64 5 20 20 old 1 1-44 3 2 1A621 1 16 thirty old 1 1-47 3 1 1A577 1 14 15 old To 1-16 2 0 1A732 2 18 9 old To 3-20 3 0 1A74 1 18 23 old 1 1-51 3 1 1A695 3 19 9 old To 1-5 1 1 1A479 1 16 25 old To 3-20 1 0 1A182 1 24 28 old To 1-5 1 0 1A693 3 9 17 old To 1D-39 2 0 1A79 0 16 thirty old 1 1-47 1 3 1A27 0 19 50 old 1-9 1 0

- 106 042696

Name At CDR3 (a.k.) SEQ ID NO (+) Length Binding NEUT # relatives 1B2434 AVYDSSLSLGL 863 0 eleven CD4BS + 7 1B218 QHRSNWPWT 864 2 9 CD4BS + 10 1B331 HQYFSTPRT 865 2 9 CORE + 4 1B2174 HQYFNTPRT 866 2 9 No data 1 1B2055 QQYEDPPWT 867 0 9 No data No data 3 1B2133 QQTYSNPRM 868 1 9 CD4i - 2 1A64 ASWDDSLSGW V 869 0 eleven CD4BS + 24 1A621 ASWDNSLSGPV 870 0 eleven CD4BS + 3 1A577 QQYNSFPPT 871 0 9 CD4BS + 8 1A732 QQYGRSP 872 1 7 CD4BS + 1 1A74 GTWDSSLSAVL 873 0 eleven CORE + 2 1A695 QQYD S 874 0 uh CORE + 2 1A479 HQYAYSPRT 875 2 9 CORE + eleven 1A182 QQYKSYSGT 876 0 9 CD4i + 3 1A693 QHSFGSPPWT 877 1 eleven CD4i - 1 1A79 AAWDD SFDYV 878 0 10 V3 + 27 1A27 QQLRT 879 1 5 GP41 - 8

Table 4A

Patient 3, Clone RU01

3BNC62 3BNC176 3BNC60 3BNC117 3BNC95 3BNC104 MW965.26 <0.09 <0.10 <0.04 <0.09 <0.07 >50 BaL.26 <0.09 <0.10 <0.04 <0.09 <0.07 0.025 DJ263.8 <0.09 <0.10 <0.04 <0.09 <0.07 0.054 6535.3 0.68 0.46 0.54 0.55 1.0 >50 RHPA4259.7 <0.09 <0.10 <0.05 0.041 <0.07 0.0252 TRO.11 <0.09 <0.10 <0.05 0.077 <0.07 3.791 PVO.4 <0.09 <0.10 0.09 <0.09 <0.07 0.348 YU2.DG <0.09 <0.10 <0.05 0.054 <0.07 0.034 3BNC91 3BNC55 3BNC89 3ANC3 3BNC53 3BNC72 MW965.26 <0.08 0.04 >0.05 0.18 0.09 <0.06 BaL.26 >178 >30 >110 >50 >30 >139 DJ263.8 >178 >30 >110 >50 >30 >139 6535.3 1 2, 6 1.7 >50 13, 6 8.49 RHPA4259.7 <0.08 2.2 12.4 7, 66 100.6 >139 TRO.11 3, 06 18.4 52.4 10.76 >155 >139 PVO.4 0.44 3, 9 2.7 36, 77 >155 >139 YU2.DG <0.08 0.9 0.39 35, 01 >155 >139 3BNC156 3BNC158 3BNC153 3BNC108 MW965.26 0.08 0.11 0.15 No data BaL.26 >111 >109 >100 20.6 DJ263.8 >111 >109 >100 >55 6535.3 11.1 9, 9 28, 9 >55 RHPA4259.7 >111 >109 >100 45, 91 TRO.11 >111 >109 >100 >55 PVO.4 >111 >109 >100 >55 YU2.DG >111 >109 >100 25.5 3BNC142 3BNC66 3BNC42 3BNC102

- 107 042696

MW965.26 0.14 1.24 No data >50 BaL.26 >172 >189 >2 6 >50 DJ263.8 >172 >189 >2 6 >50 6535.3 >172 >189 >2 6 >50 RHPA4259.7 >172 >189 >2 6 >50 TRO.11 >172 >189 >2 6 >50 PVO. 4 >172 >189 NF >50 YU2.DG >172 >189 >2 6 >50

Patient 3, Clones RU02-07

3A67 3A383 3BNC8 3ANC44 ZA576 3ANC38 MW965.26 0, 1 0.5 0.74 25.49 >50 >50 BaL.26 19, 2 5, 3 >50 27.91 27 >50 DJ263.8 >50 >50 >50 >50 >50 >50 6535.3 >50 No data >50 >50 >50 >50 RHPA4259.7 >50 No data >50 >50 >50 >50 TRO.11 >50 No data >50 >50 >50 >50 PVO. 4 >50 No data >50 >50 >50 >50 YU2.DG >50 No data >50 >50 >50 >50

Clone B12 and NIH 45

B12 VRC01 NIH45-46 MW965.26 0.2 <0.08 0.04 BaL.26 0.2 0, 1 <0.04 DJ263.8 >50 0.08 <0.04 6535.3 1.4 0.539 0.14 RHPA4259.7 0, 1 0.06 0.034 TRO.11 >50 0.2 1.9 PVO. 4 >50 0.2 0.17 YU2.DG 2.2 0.12 <0.05

Patient 1, Clone RU08

Table 4B

1B2640 1B2530 1B2364 1NC2 1NC9 1B2490 MW965.26 41.76 0.762 1.85 >50 >50 >50 BaL.26 0.08 >50 >25 0.11 1.37 0.058 DJ263.8 >50 2.71 3.75 >50 >50 >50 6535.3 >50 >50 >25 >50 >50 >50 RHPA4259.7 0.04 3, 6 2.18 0.59 0.09 0.414 TRO.11 0.23 0.516 0.27 0.17 0.2 1.06 PVO. 4 1.05 0.275 0.161 0.37 0.34 2.97 YU2.DG 0.2 0.209 2.46 0.12 0.13 0.125

- 108 042696

1B2351 1B344 1NC24 1NC3 1NC7 1NC33 MW965.26 >50 >50 >50 >25 >50 >50 BaL.26 >50 >50 >50 >25 >50 >50 DJ263.8 8.46 12, 62 >50 >25 >50 >50 6535.3 >50 >50 >50 >25 >50 22.04 RHPA4259.7 36.48 29, 98 >50 >25 34.27 >50 TRO.11 0.331 0.27 0.2 3.37 16.57 >50 PVO.4 0.25 0.27 0.19 6, 68 1.39 1.84 YU2.DG 0.058 0.25 0.16 18.26 >50 >50 1NC108 1B2644 1B2339 1NC123 MW965.26 >50 >25 >25 >50 BaL.26 >50 >25 >25 >50 DJ263.8 >50 >25 >25 >50 6535.3 >50 >25 >25 >50 RHPA4259.7 >50 >25 >25 >50 TRO.11 19.37 >25 >25 >50 PVO.4 3.13 >25 >25 >50 YU2.DG >50 >25 >25 >50

Patient 1, Clone RU09

1B218 MW965.26 >119 BaL.26 eleven DJ263.8 >119 6535.3 3, 6 RHPA4259.7 >100 TRO.11 >100 PVO.4 >100 YU2.DG >100

Table 4C

Patient 8, Clone RU10

8ANC192 8ANC134 8ANC13 8ANC131 8ANC182 8ANC50 8ANC45 MW965.26 >73 >50 >50 >50 >115 >50 >50 BaL.26 0.08 0.02 0.04 0.06 0.08 0.17 0.296 DJ263.8 <0.03 0.003 0.008 0.004 <0.05 0.04 0.041 6535.3 0.34 0.06 0.27 0.2 0.89 2.27 0.813 RHPA4259.7 >50 >50 >50 >50 >100 >50 >50 TRO.ll >100 >50 >50 >50 >100 >50 >50 PVO.4 0.89 0.46 0.63 0.81 1.2 3.89 4.259 YU2.DG 0.09 0.15 0.21 0.18 0.22 0.42 0.499

Patient 8, Clones RU11-15

8ANC57 8ANC195 8ANC24 8ANC14 8ACN5 MW965.26 24.1 >50 0.29 2.01 >50 BaL.26 4.35 >50 47.53 >50 >50 DJ263.8 30, 19 >50 >50 >50 >50 6535.3 >103 0.2 >50 >50 >50 RHPA4259.7 1.65 0.34 >50 >50 >50 TRO.ll 32.07 0.18 >50 >50 >50 PVO.4 101.15 0.52 >50 >50 >50 YU2.DG 27.52 0.79 >50 >50 >50

- 109 042696

Table 4D

Patient 12, Clone RU16

12 Al 2 12A21 12A4 12 A3 7 12A22 12Al6 MW965.26 0.042 0.075 0.098 0.056 0.06 0.167 BaL.26 0.017 <0.001 <0.001 0.005 0.04 0.042 DJ263.8 0.002 0.035 0.017 0.013 0.08 0.012 6535.3 21.97 >50 >50 >50 >25 15, 44 RHPA4259.7 0.086 0.038 0.041 0.042 0.04 0.207 TRO.11 0.288 0.164 0.257 0.827 0.56 0.751 PVO. 4 0.928 0.584 0.819 0.516 0.45 2.44 YU2.DG 0.084 0.015 0.018 0.019 0.11 0.234 12A20 12A6 12A23 12A46 12A55 MW965.26 0.192 0.112 5, 1 >50 0.58 BaL.26 0.035 0.072 0.57 0.013 2.87 DJ263.8 0.05 0.004 0.63 5, 79 >50 6535.3 48.73 >2 4 14.73 48.85 >50 RHPA4259.7 0.109 0.227 0.496 >50 >50 TRO.11 0.689 1.52 2.88 >50 21.45 PVO. 4 3, 04 3.32 2.24 2, 18 0.99 YU2.DG 0.142 0.222 0.053 0.49 0, 1

Clone B12 and NIH45

B12 VRC01 NIH45-46 MW965.26 0.2 <0.08 0.04 BaL.26 0.2 0, 1 <0.04 DJ263.8 >50 0.08 <0.04 6535.3 1.4 0.539 0.14 RHPA4259.7 0, 1 0.06 <0.05 TRO.11 >50 0.2 19 PVO. 4 >50 0.2 0.17 YU2.DG 2.2 0.12 <0.05

Table 4E

Patient 3, clone RU01

3BNC62 3BNC176 3BNC60 3BNC117 3BNC95 3BNC104 MW965.26 <0.09 <0.10 0.09 <0.09 <0.07 >50 BaL.26 <0.09 <0.10 <0.04 <0.09 <0.07 0.09 DJ263.8 0, 1 <0.10 0, 1 0, 1 0, 1 0.187 6535.3 2.24 1.7 1.77 2.44 4.5 >50 RHPA4259.7 <0.09 <0.10 0.07 0.137 <0.07 0.06 TRO.11 <0.09 <0.10 0.12 0.077 <0.07 30.847 PVO. 4 0.23 0.16 0.27 0.19 0.23 0.901 YU2.DG <0.09 <0.10 0.07 0.054 <0.07 0.097 3BNC91 3BNC55 3BNC89 3ANC3 3BNC53 3BNC72 3BNC156 MW965.26 <0.08 0.15 0.16 0.64 0.61 0.37 0.47 BaL.26 >178 >30 >110 >50 >30 >139 >111 DJ263.8 >178 >30 >110 >50 >30 >139 >111 6535.3 6.7 5.53 5.92 >50 73.38 133.665 69.66 RHPA4259.7 0.52 8.03 >110 >50 >155 >139 >111 TRO.ll 32.31 41.67 >110 >50 >155 >139 >111 PVO.4 2.65 6.5 10.18 >50 >155 >139 >111 YU2.DG <0.08 1.07 1.49 >50 >155 >139 >111

- 110 042696

3BNC158 3BNC153 3BNC108 3BNC142 3BNC66 3BNC42 3BNC102 MW965.26 0.6 0.63 No data 0.8 29.98 No data >50 BaL.26 >109 >100 >55 >172 >189 >26 >50 DJ263.8 >109 >100 >55 >172 >189 >26 >50 6535.3 97.75 >100 >55 >172 >189 >26 >50 RHPA4259.7 >109 >100 >55 >172 >189 >26 >50 TRO.11 >109 >100 >55 >172 >189 >26 >50 PVO.4 >109 >100 >55 >172 >189 No data >50 YU2.DG >109 >100 >55 >172 >189 >26 >50

Patient 3, Clones RU02-07

3A67 ZAZ 83 3BNC8 3ANC44 ZA576 3ANC38 MW965.26 16 >25 0.74 >50 >50 >50 BaL.26 >50 >25 >50 >50 >50 >50 DJ263.8 >50 >25 >50 >50 >50 >50 6535.3 >50 No data >50 >50 >50 >50 RHPA4259.7 >50 No data >50 >50 >50 >50 TRO.11 >50 No data >50 >50 >50 >50 PVO.4 >50 No data >50 >50 >50 >50 YU2.DG >50 No data >50 >50 >50 >50

Clone B12 and NIH 45

B12 VRC01 45-46 MW965.26 No data <0.08 0.21 BaL.26 No data 0, 1 0.06 DJ263.8 No data 0.553 0.06 6535.3 No data 2.7 0.28 RHPA4259.7 0.39 0.185 0.146 TRO.11 >50 0.832 9.56 PVO.4 >50 1.2 0.47 YU2.DG 7, 8 0.372 0.08

Patient 1, Clone RU08

Table 4F

1B2640 1B2530 1B2364 1NC2 1NC9 1B2490 1B2351 MW965.26 >50 >50 >2 5 >50 >50 >50 >50 BaL.26 0.32 >50 >2 5 0.51 19, 92 0.3 >50 DJ263.8 >50 >50 >2 5 >50 >50 >50 >50 6535.3 >50 >50 >2 5 >50 >50 >50 >50 RHPA4259.7 0.25 >50 >2 5 4.33 0.4 1.97 >50 TRO.11 1, 62 2.46 1.77 0.55 0.65 3.58 1.13 PVO.4 2.97 1.25 0.65 1, 08 1.32 10.57 0.88 YU2.DG 0.7 7.74 >2 5 0.39 0.56 0.59 0.48 1B344 1NC24 1NC3 1NC7 1NC33 1NC108 1B2644 MW965.26 >50 >50 >25 >50 >50 >50 >25 BaL.26 >50 >50 >25 >50 >50 >50 >25 DJ263.8 >50 >50 >25 >50 >50 >50 >25 6535.3 >50 >50 >25 >50 >50 >50 >25 RHPA4259.7 >50 >50 >25 >50 >50 >50 >25 TRO.11 0.89 0.66 >25 >50 >50 >50 >25 PVO.4 0.94 0.6 >25 7.17 10, 12 25, 08 >25 YU2.DG 1.29 0.55 >25 >50 >50 >50 >25

- 111 042696

1B2339 1NC123 MW965.26 >25 >50 BaL.26 >25 >50 DJ263.8 >25 >50 6535.3 >25 >50 RHPA4259.7 >25 >50 TRO.11 >25 >50 PVO. 4 >25 >50 YU2.DG >25 >50

Patient 1, Clone RU09

1B218 MW 9 65.26 >119 BaL.26 5, 61 DJ263.8 >119 6535.3 35, 12 RHPA4259.7 >100 TRO.11 >100 PVO. 4 >100 YU2.DG >100

Table 4G

Patient 8, Clone RU 10

8ANC192 8ANC134 8ANC13 8ANC131 8ANC182 8ANC50 8ANC45 TR. eleven >73 >50 >50 >50 >115 >50 >50 BaL.26 0.43 0.11 0.18 0.31 0.73 0.77 7.45 DJ263.8 0.1 0.044 0.069 0.046 0.11 0.15 0.166 6535.3 1.43 2 2.3 1.9 3.93 13.65 10.473 RHPA4259.7 >100 >50 >50 >50 >100 >50 >50 TR. eleven >100 >50 >50 >50 >100 >50 >50 PVO.4 3.94 2.5 3.7 4.9 4.43 14.99 17.315 YU2.DG 0.51 0.616 1.07 0.92 1.46 1.59 2.942

Patient 8, Clones RU11-15

8AN57 8AN195 8AN24 8AN14 8AN5 TRO.11 >103 >50 0.76 6, 64 >50 BaL.26 24.76 >50 >50 >50 >50 DJ263.8 >103 >50 >50 >50 >50 6535.3 >103 0.91 >50 >50 >50 RHPA4259.7 14.44 1.56 >50 >50 >50 TRO.11 >103 0.89 >50 >50 >50 PVO.4 >103 1.87 >50 >50 >50 YU2.DG 91.49 2.77 >50 >50 >50

Table 4H

Patient 12, Clone RU16

12 Al 2 12A21 12A4 12 A3 7 12A22 12Al6 MW965.26 0.2 0.85 1.24 0.3 0.21 0.58 BaL.26 0.08 0.004 0.007 0.03 0.14 0.25 DJ263.8 0.31 0.42 1.06 0.57 1.86 0.12 6535.3 >50 >50 >50 >50 >25 >42 RHPA4259.7 0.4 0.13 0.19 0.19 0.13 0.93 TRO.11 0.98 0.57 1.12 3, 81 1.94 2.57 PVO.4 3, 15 2.09 2.95 1.8 1.49 8.72 YU2.DG 0.31 0.06 0, 1 0.07 0.36 1.13

A2 0 K 12A6 12A23 12A46 12A55

- 112 042696

MW965.26 2.2 0.52 >50 >50 4.49 BaL.26 0.23 0.47 3.47 0.08 >50 DJ263.8 No data 0.08 30, 81 >50 >50 6535.3 No data >24 >50 >50 >50 RHPA4259.7 0.49 1.02 1.69 >50 >50 TRO.11 2.41 5, 15 10, 11 >50 >50 PVO. 4 11.2 17.34 7.81 797 4.3 YU2.DG 0.67 1.2 0.19 0.25 0.29

Clone B12 and NIH45

B12 VRC01 NIH45-46 MW965.26 0.2 <0.08 0.04 BaL.26 0.2 0, 1 <0.04 DJ263.8 >50 0.08 <0.04 6535.3 1.4 0.539 0.14 RHPA4259.7 0, 1 0.06 <0.05 TRO.11 >50 0.2 19 PVO. 4 >50 0.2 0.17 YU2.DG 2.2 0.12 <0.05

Table 5A

Tzm-bl in vitro neutralization assay, extended panel _IC50 values

B12 VRC01 NIH45- 46 3BNC60 3BNC62 3BNC117 3BNC55 Q842.dl2 >50 0.03 0.008 0.01 <0.01 <0.01 0.011 3415.vl.cl 2.5 0.06 0.017 0.1 0.17 0.17 0.11 3365.v2.c20 >50 0.03 0.029 0.02 0.03 0.03 0.221 H086.8* >50 >50 >30 >15 >15 >15 >30 ZM53M.PB12 >50 1.3 0.187 0.22 0.3 0.21 12.549 Dul72.17* 0.3 >50 >30 3.81 1.72 1.19 3.518 ZM109F.PB4 >50 0.128 0.059 0.22 0.14 0.14 0.083 3016.v5.c45 1.1 0.16 >30 1.4 0.42 1.38 >30 231965.cl 0.07 0.34 0.021 0.07 0.05 0.05 0.505 X1254_c3 >50 0.07 0.027 0.09 0.08 0.08 0.138 250-4* >50 >50 >30 >15 >15 >15 0.236 251-18 >50 2.5 1.445 0.35 0.32 0.26 >30 278-50* >50 >50 >30 >15 >15 >15 >30 620345.cl* >50 >50 >30 >15 >15 >15 >30 R1166.C1 >50 1.7 0.445 0.14 0.32 0.17 0.298 1NC9 1B2530 8ANC131 8ANC134 8ANC195 12A12 12A21 Q842.dl2 0.02 0.249 0.053 0.061 >30 0.014 0.015 3415.vl.cl 0.266 0.065 0.299 0.323 2.404 0.121 0.82 3365.v2.c20 0.329 4.357 >30 >30 >30 0.068 0.045 H086.8* >30 >30 >50 >50 0.095 >30 >30 ZM53M.PB12 0.705 0.912 >30 >30 9.626 0.593 0.42 Dul72.17* >30 >30 >30 >30 10.797 0.196 0.126 ZM109F.PB4 0.023 >30 >30 >30 >30 0.148 2.104 3016.v5.c45 >30 >30 >30 >30 0.195 1.163 0.097 231965.cl 0.393 0.168 6.346 >30 0.514 2.217 >30 X1254_c3 >30 >30 >30 >30 1.524 1.032 26.793 250-4* >30 >30 >50 >50 >50 >30 >30 251-18 1.234 9.847 0.968 1.56 0.284 2.622 1.713 278-50* >30 >30 >50 >50 >50 >30 >30 620345.cl* >30 >30 >50 >50 >50 >30 >30 R1166.C1 0.651 0.119 >30 >30 0.986 0.342 0.292

- 113 042696

Table 5B

In vitro Tzm-bl neutralization assay, extended panel IC ₈₀ values

B12 VRC01 45-46 3BNC60 3BNC62 3BNC117 3BNC55 Q842.dll >50 0.096 0.026 0.03 0.03 0.01 0.062 3415.vl.cl 14.1 0.15 0.069 0.37 0.4 0.47 0.388 3365.v2.c20 >50 0.17 0.114 0.08 0.09 0.1 2.341 H086.8* >50 >50 >30 >15 >15 >15 >30 ZM53M.PB12 >50 4 0.652 0.76 1.1 0.85 >30 Dul72.17* 2.6 >50 >30 >15 12.18 8.9 >30 ZM109F.PB4 >50 0.754 0.22 1.23 0.78 0.88 0.396 3016.v5.c45 4 0.42 >30 7.38 2.35 >15 >30 231965.С1 0.16 1.2 0.1 0.25 0.22 0.22 2.78 X1254 sz >50 0.19 0.078 0.29 0.27 0.27 0.571 250-4* >50 >50 >30 >15 >15 >15 1.922 251-18 >50 H.2 5.255 0.96 1 0.82 >30 278-50* >50 >50 >30 >15 >15 >15 >30 620345.С1* >50 >50 >30 >15 >15 >15 >30 R1166.C1 >50 4.6 1.679 0.51 0.89 0.64 2.351 1NC9 1B2530 8ANC131 8ANC134 8ANC195 12A12 12A21 Q842.dl2 0.133 2.191 0.179 0.205 >30 0.06 0.066 3415.vl.cl 1.002 0.35 1.555 2.643 17.743 0.418 0.296 3365.v2.c20 2.163 >30 >30 >30 >30 0.192 0.166 H086.8* >30 >30 >50 >50 5.328 >30 >30 ZM53M.PB12 2.771 4.022 >30 >30 >30 2.069 1.458 Dul72.17* >30 >30 >30 >30 >30 0.992 0.637 ZM109F.PB4 0.146 >30 >30 >30 >30 0.698 13.686 3016.v5.c45 >30 >30 >30 >30 0.872 11,864 0.358 231965.C1 2.276 0.963 >30 >30 2.355 15.102 >30 X1254_c3 >30 >30 >30 >30 6.949 5.777 >30 250-4* >30 >30 >50 >50 >50 >30 >30 251-18 6.291 >30 5.55 6.281 1.511 9.39 6.063 278-50* >30 >30 >50 >50 >50 >30 >30 620345.C1* >30 >30 >50 >50 >50 >30 >30 R1166.C1 2.669 0.684 >30 >30 4.83 1.85 2.137

- 114 042696

Table 6

Affinity of IgG antibodies to YU-2 ligands gp140 and 2CC-core measured by surface plasmon resonance

gp!40 2SS-Core cDM^C ¹ ) k _d (with ¹ ) K _D (M) ka (MV) ka (from ¹ ) K _D (M) 12А12 4.59E+04 1.44Е-05 ZD5E-10 6,33E+04 1.70E-06 2.69Е-11 12A21 9D8E+04 ZD4E-07 3.75Е12 1.82E+05 3,ZOE-04 1.81E-09 12AGL / / / / / / 3BNC60 2.73E+04 1.86Е-04 6.81Е-09 3.02E+04 1.64Е-03 5.45Е-08 3BNC117 3.04E+04 1.99E-04 6.54Е-09 1.49Е-03 4.05E+04 3.68E-08 3BNC55 1D1E+04 7D5E-04 5.78Е-08 8D5E-04 ZD6E+04 2.57Е-08 3BNC66 1.60E+04 1.41Е-03 8.81Е-08 3.96E+04 1,ZZE-OZ 3.36E-08 3BNC156 1DZE+04 1.98E-03 1.75Е-07 1.88E+04 1.53Е-03 8D2E-08 3BNC108 / / / / / / 3BNC60GL / / / / / / 8ANC131 6D9E+04 1.09E-03 1.65Е-08 4.88E+04 3.23E-03 6.61E-08 8ANC134 1.55E+04 1.74Е-03 1DZE-07 2.08E+04 9.57Е-04 4.61E-08 8AGL / / / / / / 8ANC195 4.88E+04 1.67Е-03 3.43Е-08 2.41E+04 1.32E-03 5.47Е-08 1NC9 4.83E+04 5.81E-04 1.20E-08 5D1E+04 2D6E-04 4.61E-09 1B2530 4.74E+04 1.62E-03 3.42E-08 6.83E+04 4.02E-04 5.90E-09 1GL / / / / / / 4546 4.26E+04 2.87Е-04 6.75Е-09 1D2E+05 4.94Е-04 4.40E-09 VRC01 1.83E+04 8.08Е-06 4.41Е-10 2.84E+04 3.25E-05 1D5E-09

Table 7A

Substitution ratio/silent mutations for heavy chain sequences of 10 selected antibodies

All nucleotides Consensus nucleotides non-consensual nucleotides 3BNC117HC 1.8 1.0 3.5 3BNC60HC 2.0 1.1 4.4 12A12NS 2.8 1.7 6, 3 12A21NS 2, 6 1.5 4.8 NIH4546HC 1.7 0.9 5, 5 VRCO1HC 2.2 1.1 22.0 8ANC131HC 2.7 1.3 8.0 8ANC134HC 2.2 1.5 3.7 1V2530NS 2.0 0.9 11.0 1NC9HC 1.9 0.7 12.0

Table 7B

Substitution ratio/silent mutations for light chain sequences of 10 selected antibodies

All nucleotides Consensus nucleotides non-consensual nucleotides 3BNC117KC 1.7 0.8 2, 8 3BNC60KC 1.7 0.7 4.0 12A12KS 1.7 0.6 4.0 12A21KS 2.5 1.4 4.3 NIH4546KC 1.7 0.9 thirty VRCO1KC 1.8 0.8 4.0 8ANC131KC 1.5 0.5 4.2 8ANC134KC 1.5 0.5 4.2 1B2530LC 1.9 20 18 1NC9LC 1.2 0.9 18

- 115 -

Claims (22)

Table 8 Collection of data on crystallization and refinement of statistics Crystal Fab 3BN60 Data collection* Wavelength (A) 0.9537 Symmetry group R21 Unit cell dimensions a (A) 63.6 b (A) 155.7 s (A) 74.8 α,β,γ (Υ) 90.0, 110.4, 90.0 Resolution, (A) 39.172.65 Rmrgd-F ( % ) ^§ 8.3 (55.5) Rmeas ( ) § 7.7 (53.4) Ι/σΙ 15.7 (2.5) Completion (%) 96.0 (68.1) multiplicity 5.0 (3.6) Reflections 192709 Unique Reflections 38111 Clarification Resolution (A) 39.172.65 Number of reflections 37086 Rwork/Rfree (%) ⁺ 20.7/25.7 RMSD bond lengths (A) 0.01 RMSD bond angles (Y) 1.3 Average B-factor A ² 64.9 Ramachandran's analysis Favorable (%) 91.9 Permissible (%) 7, 6 Emissions (%) 0.5 CLAIM 1. An isolated anti-HIV antibody or antigen-binding fragment thereof, containing the CDR1, CDR2 and CDR3 regions in the specified variable region of the heavy chain, which are NCPIN, WLKPRGGAVNYARKFQG and GKYCTARDYYNWDFEH, respectively; and CDR1, CDR2, and CDR3 regions in said light chain variable region, which are RTSQSGSL, SGSTRAA, and QQYEF, respectively. 2. The isolated anti-HIV antibody or antigen-binding fragment thereof according to claim 1, wherein the anti-HIV antibody or antigen-binding fragment neutralizes the HIV virus SM53M.PB12 at an IC ₅ o concentration of less than 1.0 μg/ml, or the HIV R1166 virus .c1 at an IC ₅ o concentration of less than 1.0 µg/mL. 3. An isolated anti-HIV antibody or antigen-binding fragment thereof according to claim 1, wherein the anti-HIV antibody or antigen-binding fragment thereof neutralizes a VRC01-resistant HIV virus at an IC ₅₀ concentration of less than 30 μg/ml. 4. The isolated anti-HIV antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein the antibody is a chimeric antibody, a humanized antibody, a recombinant antibody, or a human antibody. 5. The isolated anti-HIV antibody or antigen-binding fragment thereof according to claim 4, wherein the antibody is a human antibody. 6. The isolated anti-HIV antibody or antigen-binding fragment thereof according to claim 4, wherein the antibody is a recombinant antibody. 7. Composition for the prevention or treatment of HIV infection or a disease associated with HIV, containing an isolated anti-HIV antibody or an antigen-binding fragment according to any one of claims 1 to 6 and a second antibody other than an anti-HIV antibody, or an antigen-binding fragment thereof . 8. A nucleic acid molecule encoding an isolated anti-HIV antibody or an antigen-binding fragment thereof according to any one of claims 1-6. 9. A vector containing a nucleic acid molecule according to claim 8. - 116 042696 10. A cell for expressing an anti-HIV antibody or an antigen-binding fragment thereof, containing the vector according to claim 9. 11. A pharmaceutical composition comprising at least one antibody or antigen-binding fragment thereof according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier. 12. Pharmaceutical composition according to claim 11, further comprising a second therapeutic agent. 13. The pharmaceutical composition of claim 12, wherein said second therapeutic agent is an antiviral agent. 14. The pharmaceutical composition according to any one of claims 11 to 13, wherein the anti-HIV antibody or antigen-binding fragment thereof is a recombinant antibody or antigen-binding fragment thereof. 15. The use of a pharmaceutical composition according to any one of claims 11 to 14 for the prevention or treatment of HIV infection or an HIV-related disease. 16. A method for producing an anti-HIV antibody or antigen-binding fragment according to claims 1 to 6, wherein the method includes culturing a cell containing a vector containing a nucleic acid encoding the heavy and light chains of said antibody or fragment under nucleic acid expression conditions; and isolating said anti-HIV antibody or fragment thereof. 17. A method for detecting an anti-HIV antibody or antigen-binding fragment thereof according to any one of claims 1 to 6 in a patient, wherein the method comprises contacting a biological sample obtained from the patient with an agent that binds to said antibody or antigen-binding fragment thereof, or which hybridizes to DNA or mRNA encoding an antibody or fragment; and analyzing the biological sample for the presence of an anti-HIV antibody or antigen-binding fragment thereof, or a cell that contains at least one DNA or mRNA encoding the antibody or antigen-binding fragment thereof. 18. A kit for the prevention or treatment of an HIV infection or an HIV-related disease, comprising a pharmaceutically effective amount of at least one isolated anti-HIV antibody or antigen-binding fragment thereof according to any one of claims 1 to 6 and a pharmaceutically effective amount of an anti-HIV agent selected from the group consisting of a non-nucleoside reverse transcriptase inhibitor, a protease inhibitor, an entry or fusion inhibitor, and an integrase inhibitor. 19. The kit of claim 18, wherein the isolated anti-HIV antibody or antigen-binding fragment thereof and the anti-HIV agent may optionally take the form of a single pharmaceutical composition. 20. A kit for diagnosing, predicting, or monitoring HIV treatment in a patient, comprising one or more detection reagents that specifically bind to an anti-HIV antibody according to any one of claims 1-6 in a biological sample of an individual. 21. A kit according to claim 20, additionally containing reagents for PCR. 22. The kit according to claim 20, additionally containing reagents for mass spectrometry. - 117 -