Anti-monomethyl aurestatin antibodies and antibody fragments

Abstract

The present invention provides compositions and methods for reducing off-target toxicity of antibody-drug conjugates (ADCs). The compositions comprise an ADC and an agent targeting a drug (payload) delivered by or derived from the ADC. The ADC and the agent targeting the payload may be delivered together or separately when the ADC treats various conditions (e.g., tumors). Examples of agents targeting a payload include antibodies, fragments thereof, or modifications thereof.

Description

[0001] CROSS REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 373,367 filed on August 24, 2022 and U.S. Provisional Application No. 63 / 520,689 filed on August 21, 2023, the entire contents of each application are incorporated herein by reference.

[0003] Sequence Listing

[0004] This application contains a sequence listing that has been submitted in .xml format, the entire contents of which are hereby incorporated by reference. The .xml copy was created on August 24, 2023, is named "011520_01784_Balthasar_PCT.xml" and is 183,732 bytes in size. Background Art

[0005] Anti-cancer antibody-drug conjugates (ADCs) are used to deliver drugs (which may be toxins or other cell growth inhibitors (referred to herein as drugs or payload molecules)) to cancer cells. Currently, 12 ADCs are available in the United States, and about 100 ADCs are under development (Chau et al., Lancet. 2019; 394(10200): 793-804; Coats et al., Clin Cancer Res. 2019. Epub 2019 / 04 / 14.doi: 10.1158 / 1078-0432.CCR-19-0272; Wolska-Washer et al., Drug Saf. 2019; 42(2): 295-314; Beck et al., Nat Rev Drug Discov. 2017; 16(5): 315-37). However, the clinical application of ADCs has been somewhat disappointing; many ADCs have failed in clinical trials due to severe off-target toxicity, which makes the tolerable dose lower than the level required to eradicate the tumor (Coats et al., Clin Cancer Res. 2019. Epub 2019 / 04 / 14. doi: 10.1158 / 1078-0432. CCR-19-0272; Kim et al., Biomol Ther (Seoul). 2015; 23(6): 493-509; de Goeij et al., Curr Opin Immunol. 2016; 40: 14-23; Khera et al., BioDrugs. 2018; 32(5): 465-80). The early promise of ADCs has not been fully realized due to the associated toxicity to non-target sites, so new methods need to be continuously developed in the field of cancer treatment to minimize the off-target toxicity of therapeutic payload drug molecules without compromising their anti-tumor efficacy. Summary of the invention

[0006] The present disclosure provides compositions and methods for reducing ADC off-target toxicity. For example, the present compositions and methods can be used to treat tumors with ADC while reducing ADC off-target toxicity. The drug in ADC may be referred to as "payload" herein. The composition includes an ADC and a pharmaceutical agent that targets ADC delivery or a payload derived from ADC. The pharmaceutical agent targeting the payload is referred to as "payload binder" or PBA herein. ADC and payload binders may be provided in the same composition or in different compositions. Payload binders may be peptides or antibodies or fragments or antibody mimetics or modifications thereof for ADC payloads, and they are combined with payloads. If the payload binder is an antibody or a fragment or modification thereof, it may be referred to as "anti-payload antibodies".

[0007] In one aspect, the present disclosure provides a method of inhibiting or preventing the growth of one or more tumors, comprising administering an ADC and a payload binder to an individual in need of treatment, wherein the payload binder has a specific affinity for the ADC payload. The ADC and payload binder can be administered in the form of the same composition or different compositions, by the same route or by different routes, or using the same regimen or different regimens.

[0008] In one aspect, the present disclosure provides peptides or antibodies or antibody fragments or modifications that are specific to ADC payload molecules. Anti-payload antibodies can be whole immunoglobulin molecules, such as polyclonal or monoclonal antibodies or chimeric antibodies including humanized antibodies. Antibody fragments or modifications can be antigen-binding fragments thereof, including but not limited to Fab, F(ab'), F(ab')2, Fv, dAb, Fd, CDR fragments, single-chain antibodies (scFv), bivalent single-chain antibodies, single-chain phage antibodies, double antibodies or single-domain antibodies (nano antibodies), etc. Antibody mimics can include affibodies, nanobodies, etc. Antibody fragments can be produced by synthesis, or by enzymatic or chemical cleavage of complete immunoglobulins, or can be genetically engineered by recombinant DNA technology. These techniques are well known in the art. Antibodies or their fragments or modifications can be modified to confer longer half-life, stability, etc. In one embodiment, the present disclosure provides antibodies or fragments or derivatives thereof directed against several ADC payloads, including full-length antibodies, scFvs, Fabs and other fragments specific for auristatins (e.g., monomethyl auristatin E [MMAE], monomethyl auristatin F [MMAF], monomethyl auristatin F methyl ester, monomethyl auristatin D, etc.). In a non-limiting embodiment, the anti-payload specifically binds to MMAE and / or MMAF. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 :The top 5 scFv clones identified after bio-panning and screening of immune mouse scFv phage library are provided. The binding score represents the ELISA signal of the phage bound to MMAF-biotin-streptavidin fixed on the ELISA plate for each clone (whether MMAE or T-vc-MMAE is added). The bars represent the average of the replicates with standard deviation error bars. The binding scores of clones 1B3, 1H2, 3B8, 2E8 and 2C2 decreased by> 50% after adding 30-100nM free MMAE, while there was no change in binding after adding 100nM T-vc-MMAE.

[0010] Figure 2: The sequences of the first five clones identified from the first panning are provided. The complementary determining regions identified using the Northern definition are shown in the boxed area. Clone 1H2 has Figure 1 The sequence in the top row is SEQ ID NO:31.

[0011] Figure 3 : From clones 1B3, 1H2 and 3B8 ( Figure 1 and 2 ) Random mutagenesis scFv library was constructed to identify clones with increased MMAE binding affinity. Shown are the fractional binding signals of four 96-well plates from two different panning dissociation steps (3 hours and 24 hours). The fractional binding signal of the 3-hour plate is the signal observed after adding 10 nM free MMAE divided by the control well without MMAE. The fractional signal of the 24-hour plate is the result after adding 1 nM free MMAE.

[0012] Figure 4 :right Figure 3 The clones of the screening results were sequenced. The parental scFv sequences (1B3, 1H2, 3B8) used to construct the mutagenesis library are provided, wherein 1H2 is set as the reference sequence. The complementary determining regions identified using the Northern definition are shown in the boxed area. The amino acid sequence of the variable domain of clone MA24E2 was used to guide the development of the humanized variant ABC3315. The sequence of the 1B3 construct in the top row is SEQ ID NO:31.

[0013] Figure 5 : Binding of ABC3315 to MMAF-biotin-streptavidin decreases with increasing concentrations of free MMAE and MMAF (IC50: ~1 nM), while trastuzumab-vc-MMAE (T-vc-MMAE) does not compete for binding. Points represent the mean of three replicates with standard deviation error bars.

[0014] Figure 6 : Kinetic titration binding SPR sensorgrams of ABC3315 with MMAF-peg11-biotin neutravidin (left), MMAE with ABC3315-peg12-biotin-streptavidin (middle), and polatuzumab vedotin (PV) with ABC3315-peg12-biotin-streptavidin (right) are shown. The best fit values ​​of the association rate constant (Kon), dissociation rate constant (koff), and equilibrium dissociation rate constant (KD) are provided in the inset. No binding signal was observed for PV with ABC3315-peg12-biotin-streptavidin.

[0015] Figure 7: ABC3315 selectively inhibits MMAE toxicity. (A) RAMOS cells were incubated with MMAE (10pM–100nM) with or without 500nM ABC3315. ABC3315 increased the IC50 of free MMAE by 800-fold. (B) Polatuzumab vedotin (PV), a clinically approved anti-CD79b ADC incorporating MMAE as a payload molecule, was incubated with CD79b+ Ramos cells at concentrations of 3pM–30nM with or without 500nM ABC3315. ABC3315 did not alter the on-target cytotoxicity of PV (IC50: 0.12nM alone vs 0.13nM with ABC3315). (C) ABC3315 (500nM) increased the IC50 of free MMAE against SKBR3 cells by more than 500-fold. (D) HER2+SKBR3 cells were incubated with T-vc-MMAE (with or without anti-MMAE fab) with negligible changes in cytotoxicity (IC50: 0.03 nM alone vs 0.04 nM ABC3315). Points represent the mean of triplicate wells with standard deviation error bars.

[0016] Figure 8 ABC3315 did not alter the efficacy of PV. 3 Nu / J mice were randomly assigned to receive PBS, 1 or 3 mg / kg PV (with or without 12-fold molar excess of ABC3315) (n=8 / group). (A) Changes in tumor volume over time for each group are shown with standard deviation error bars. 3 (B) The survival probability of each group is presented over time. According to the Log-Rank test, the combination treatment of ABC3315 did not significantly change the survival rate of mice treated with PV at a dose of 1 mg / kg (p=0.075) or 3 mg / kg (p=0.89).

[0017] Fig. 9: ABC3315 reduces body weight loss in mice treated with 120 mg / kg PV. Swiss Webster mice were injected intravenously with 120 mg / kg PV and PBS or a 3-fold molar excess of ABC3315 (n=5 / group). (A) Body weight loss over time for each group is presented; points represent mean percent change in body weight and error bars represent standard deviation. (B) Mean percent body weight change from the lowest point in each group is presented with standard deviation error bars. The group nadir body weight loss (nadir body weight loss) of mice administered ABC3315 with PV was significantly reduced from an average of 11.9±7.0% in mice treated with PV+PBS to 4.1±2.1% in mice treated with PV+ABC3315 (p=0.045).

[0018] Fig.10 : Evaluation of ABC3315-WEDD HSA binding. Graphs show ABC3315-WEDD injected onto an SPR chip with immobilized human serum albumin over a wide range of concentrations. Top: Observed binding and dissociation curves were fitted to a 1:1 Langmuir binding model to obtain the association rate constants. (Bottom) The observed Rmax for each concentration was used to estimate the equilibrium dissociation rate constant. The fitted values ​​of the binding parameters are provided in the inset of each panel.

[0019] Fig.11 : Evaluation of ABC3315-WE human serum albumin (HSA) binding. ABC3315-WE was injected onto an SPR chip with immobilized human serum albumin over a wide range of concentrations. Top: Observed binding and dissociation curves were fitted to a 1:1 Langmuir binding model to obtain the association rate constants. (Bottom) The observed Rmax for each concentration was used to estimate the equilibrium dissociation rate constant. The fitted values ​​of the binding parameters are provided in the inset of each panel.

[0020] Fig.12 : Affinity matured scFv sequences using ABC3315 as reference sequence. The sequence in the top row is SEQ ID NO: 31.

[0021] Fig.13 : ELISA comparison of ABC3315 mutants. The retained binding activity of ABC3315 mutants was evaluated using an indirect ELISA method. Except for VH_F37V, all mutants showed similar binding signals to MMAF-peg11-biotin-neutravidin at the Fab concentrations tested.

[0022] Fig.14Competitive cytotoxicity assay. The inhibition of MMAE-mediated cytotoxicity by the VH_F27L mutant was evaluated compared to ABC3315. Higher cell viability was observed with VH_F27L co-treatment compared to ABC3315 co-treatment over a wide range of MMAE concentrations.

[0023] Fig.15 : A graph showing free MMAE concentrations in plasma and erythrocytes after administration of 100 mg / kg TvcMMAE alone or in combination with ABC3315. ABC3315 reduced free MMAE concentrations in plasma by 85% and in erythrocytes by 73%.

[0024] Fig.16 : A graph showing the results of administering 80 mg / kg doses of TvcMMAE alone or in combination with ABC3315. Compared with mice administered with PBS vehicle, administration of TvcMMAE alone resulted in significant reductions in white blood cell counts and red blood cell counts. Compared with control mice, there was no significant reduction in white blood cells or red blood cells in mice administered with TvcMMAE in combination with ABC3315.

[0025] Fig.17 : Data showing the effect of ABC3315 on the efficacy of TvcMMAE in a HER2+ / HER2- bystander xenograft model were evaluated. No significant differences were observed between mice administered 3 mg / kg TvcMMAE alone or in combination with ABC3315. Representative tumors are provided on the right.

[0026] Fig.18 : A graph showing that ABC3320 reduces the toxicity of MMAE ADC to differentiated neutrophils. Human peripheral blood mononuclear cells were incubated with trastuzumab-vc-MMAE (TvcMMAE) alone or in combination with ABC3320. Flow cytometry was used to assess the number of neutrophils after TvcMMAE treatment, using CD66b+ as a neutrophil marker. ABC3320 increased the IC50 of TvcMMAE from 1.9nM to 27nM.

[0027] Fig.19 : The graph shows that ABC3320 co-treatment reduced the body weight loss observed in rats treated with 25 mg / kg Trastuzumab-vc-MMAE (ADC) compared to rats treated with TvcMMAE alone.

[0028] Fig. 20 : The graph shows that ABC3320 reduced hematological toxicity in rats after a 25 mg / kg dose of TvcMMAE.

[0029] Fig.21: The graph shows that ABC3320 reduces trastuzumab-vc-MMAE (25 mg / kg)-mediated hepatotoxicity in rats.

[0030] Fig. 22 : The graph shows that ABC3320 does not reduce the efficacy of polatuzumab vedotin (PV) in mice bearing Ramos xenografts, nor does it reduce the efficacy of trastuzumab-vc-MMAE (TvcMMAE) in mice bearing a mixed xenograft model of HER2+ NCI-N87 cells and HER2- MCF7 cells.

[0031] Fig.23 : Graph showing that ABC3320 reduces free MMAE concentration in plasma after administration of TvcMMAE. Swiss-Webster mice were intravenously injected with a 25 mg / kg dose of TvcMMAE in combination with phosphate buffered saline (labeled as PBS) or with ABC3320 (98 mg / kg, of which 65.3 mg / kg was co-administered with TvcMMAE at time = 0 and 32.7 mg / kg was administered at 24 hours). Mice were sacrificed at a series of time points and whole blood was collected. Plasma and red blood cells were separated by centrifugation. Plasma samples (100 μl) were placed in a rapid equilibrium dialysis device (Thermo Scientific TM , 90006) in the hole, and according to the manufacturer's advice, free MMAE was separated from bound MMAE (plasma protein binding or ABC3320 binding). Free and bound MMAE concentrations were determined using LC-MS / MS. Compared with samples obtained from mice using TvcMMAE and PBS, the MMAE free concentration of samples obtained from mice treated with ABC3320 was 200 times lower on average. For mice administered ABC3320, an increase in MMAE binding concentration in plasma was observed, which is consistent with pharmacokinetic expectations (wherein binding to ABC3320 is expected to reduce the apparent distribution volume of released MMAE). DETAILED DESCRIPTION

[0032] The present disclosure provides compositions and methods for treating diseases (e.g., cancer) using ADC, while reducing off-target toxicity associated with ADC. The method includes administering ADC and one or more payload binders for ADC drug moieties to individuals in need of treatment. The payload binder can effectively reduce the non-target toxicity of ADC or the non-target toxicity derived from the free drug dissociated therefrom. The composition includes antibodies (including fragments or modifications thereof) for ADC drugs. A composition comprising ADC and a medicament (e.g., an antibody) for a drug constituting ADC is also provided. The medicament is bound to the free drug, and when the drug is part of the ADC, it may or may not be bound to the drug. The present disclosure relates to PCT application PCT / US2020 / 063453 and publications thereof, which were published on June 10, 2021 as PCT publication WO 2021 / 113740, the entire contents of which are incorporated herein by reference.

[0033] ADC includes an antibody group, a linker group, and a drug group. The antibody group targets an antigen, such as a tumor cell antigen, and the linker group is used to connect the drug group to the antibody group, which is a drug that is cytotoxic to the target cell. The antibody group can be called an antibody or a name for an antibody. Similarly, the drug group can be called a drug or a name for a drug.

[0034] The term "treating" as used herein refers to reducing or delaying the presence of one or more symptoms or features associated with the specific condition being treated. Treatment does not necessarily imply a complete cure, nor does it preclude relapse, but can be used in connection with any such relapse.

[0035] The term "therapeutically effective amount" as used herein refers to an amount sufficient to achieve the intended purpose of treatment in a single dose or multiple doses. The exact amount required or desired will vary depending on the mode of administration, patient specific circumstances, etc. A person of ordinary skill in the art (e.g., a clinician) can determine an appropriate effective amount using this disclosure.

[0036] When a range of values ​​is provided in the present disclosure, it is to be understood that unless expressly stated otherwise, each intermediate value (to one-tenth of the lower limit of the range) and any other intermediate values ​​and ranges within the described range are included in the present disclosure. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges encompassed by the present disclosure.

[0037] As used in this disclosure, the singular includes the plural and vice versa, unless the context clearly indicates otherwise. Unless clearly indicated otherwise, the indefinite articles "a" and "an" used in the specification and claims should be understood to mean "at least one".

[0038] General references to antibodies in this disclosure are also intended to include all full-length antibodies, antibody fragments containing an antigen binding domain, and modified antibodies or fragments containing substitutions or modifications of amino acid residues, and include antibody fragments (whether modified or not) that can be linked together by covalent bonds (with or without linkers). When referring to an antibody "targeting a payload" or "directed to a payload" or similar phrases, this means that the antibody has a specific affinity for the payload when the payload is in free payload form, including when it is cleaved from the ADC. When the payload is bound to the antibody portion of the ADC, the anti-payload antibody may or may not bind to the payload.

[0039] The terms "off-target" and "non-target" in relation to toxicity refer to toxicities that accompany the administration of many chemotherapeutic agents. Although the intended purpose of administering a chemotherapeutic agent is to reduce or inhibit the growth of a tumor or any accompanying metastases, the growth, function and / or physiology of normal cells are often adversely affected during cancer treatment. Reduction of off-target or non-target toxicity is intended to reduce any adverse effects on non-tumor or non-metastatic cells.

[0040] The term "payload binder" (PBA) used in the present disclosure refers to an agent that specifically binds to the payload (drug) portion of an ADC. The PBA can be an antibody, a fragment or modification thereof, a peptide, an aptamer, a Spiegelmer, a fibronectin, a DARPin, a cyclodextrin or an affitin. When the PBA is an antibody, the PBA can be referred to herein as an anti-drug antibody or an anti-payload antibody. The anti-payload antibody can be a whole immunoglobulin molecule, such as a polyclonal or monoclonal antibody or a chimeric antibody (including a humanized antibody), or it can be an antigen-binding fragment thereof, including but not limited to Fab, F(ab'), F(ab')2, Fv, dAb, Fd, CDR fragments, single-chain antibodies (scFv), bivalent single-chain antibodies, single-chain phage antibodies, double antibodies, single-domain antibodies (nano antibodies), etc. Antibody fragments can be produced by synthesis, can be produced by enzymatic or chemical cleavage of intact immunoglobulins, or can be genetically engineered by recombinant DNA technology. These techniques are well known in the art.

[0041] The term "chimeric antibody" refers to an antibody having framework residues from one species (e.g., humans) and complementary determining regions (CDRs) from another species (which typically confer antigen binding), such as a mouse antibody that specifically binds to a payload. In a chimeric antibody, some portions of the heavy and / or light chains may be identical or homologous to sequences from a particular species, while other portions may be identical or homologous to sequences from different species. Chimeric antibodies typically exhibit reduced immunogenicity and increased stability. Techniques for cloning mouse immunoglobulin variable domains are known in the art - for example, see Orlandi et al., Proc. Natl Acad. Sci. USA 86:3833 (1989), and Leung et al., Hybridoma 13:469 (1994). As an example of a chimeric antibody, a polynucleotide encoding the variable domain of a light chain or heavy chain of an antibody from an animal other than human (e.g., mouse, rat, or chicken) can be linked to a polynucleotide encoding the constant domain of a light chain or heavy chain derived from a human antibody to produce a polynucleotide (e.g., DNA) encoding a chimeric antibody.

[0042] "Human" antibodies (also called "fully human" antibodies) are antibodies that include human framework regions and all CDRs from a single or different human immunoglobulin. Thus, a framework from one human antibody can be designed to include CDRs from different human antibodies. Methods for generating human antibodies are known in the art - for example, see Mancini et al., 2004, New Microbiol. 27:315-28; Conrad and Scheller, 2005, Comb. Chem. High Throughput Screen. 8:117-26.

[0043] "Humanized antibodies" are usually human antibodies that have one or more amino acid residues imported (i.e., introduced) from non-human sources. For example, a humanized antibody is a recombinant protein in which the CDRs of antibodies from species such as rodents, rabbits, dogs, goats, or horses are introduced into human heavy and light chain variable domains. The constant domains of antibody molecules (also called framework regions) are usually the same as those of human antibodies. The non-human immunoglobulin providing the CDRs may be referred to as "donors," and the human immunoglobulins providing the frameworks may be referred to as "acceptors." For example, in humanized immunoglobulins, all CDRs may come from donor immunoglobulins. Constant regions do not always have to be present, but if present, they may be substantially identical to human immunoglobulin constant regions, i.e., at least about 85-90%, such as about 95% or more identical. Humanized antibodies bind the same payload as the donor antibodies providing the CDRs. The acceptor framework of a humanized immunoglobulin or antibody may have a limited number of amino acid substitutions (taken from the donor framework). Humanized or other monoclonal antibodies may have additional conservative amino acid substitutions that have little effect on antigen binding or other immunoglobulin functions. Humanized immunoglobulins can be constructed by genetic engineering (see, e.g., U.S. Pat. No. 5,585,089 and U.S. Publication No. 2010 / 0196266). For example, a murine monoclonal antibody can be isolated or generated and then humanized.

[0044] Antibody fragments can be produced by enzymatic digestion. For example, papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, and an "Fc" fragment. The Fab fragment contains the entire L chain and the variable region domain of the H chain (VH), and the first constant domain of one heavy chain. Each Fab fragment is monovalent in terms of antigen binding, that is, it has a single antigen-binding site. Pepsin treatment of antibodies produces a single large F(ab')2 fragment, which roughly corresponds to two disulfide-linked Fab fragments with divalent antigen-binding activity and is capable of cross-linking antigens. "Fv" is the smallest antibody fragment containing a complete antigen recognition and binding site, and a single-chain Fv, also abbreviated as "sFv" or "scFv", is an antibody fragment containing VH and VL antibody domains connected to a single polypeptide chain. The term "diabody" refers to a small antibody fragment prepared by constructing an sFv fragment with a short linker between the VH and VL domains, thereby achieving interchain pairing of the V domains rather than intrachain pairing, thereby producing a bivalent fragment, i.e., a fragment with two antigen-binding sites. Single domain antibodies (sdAbs) are antibody fragments with a single monomeric variable antibody domain. sdAbs can be made from heavy chain antibodies in camelids. Antibody fragments can be single variable regions or peptides consisting of or comprising a single CDR. Single chain antibodies have a heavy chain variable domain and a light chain variable domain linearly connected by a joint. A polynucleotide (e.g., DNA) encoding a single chain antibody can be produced by combining a polynucleotide encoding a heavy chain variable domain, a polynucleotide encoding a joint (usually 10-20 nucleotides) and a polynucleotide encoding a light chain variable domain, wherein the heavy chain variable domain and the light chain variable domain are both derived from human antibodies.

[0045] Antibodies that can be used for the present method can be obtained from humans or non-human animals. Antibodies can be of any class (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). In one embodiment, single-domain antibodies or nanobodies produced by camelids in response to the introduction of APP cleavage products (or its peptide fragments) into camelids can be used. Nanobodies are usually heavy chain antibodies, and therefore contain heavy chain homodimers, without antibody light chains. These antibodies generally contain a single variable domain and two constant domains (CH2 and CH3).

[0046] The disclosure also provides sequences having homology to proteins or peptide sequences described herein (including antibody sequences). In various embodiments, the homologous sequences have at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to proteins or peptide sequences disclosed herein.

[0047] The payload molecule can be a drug molecule that causes cytotoxicity. For example, any molecule for treating cancer can be used. Examples include compounds, DNA, RNA, peptides, etc. In one embodiment, the drug molecule can be covalently bound to the N-terminal amino acid of the antibody through a reactive group or through a joint. Examples of reactive groups cross-linked with the N-terminal α-amine group of an antibody (such as a light chain or a heavy chain) include isothiocyanates, isocyanates, acyl azides, NHS esters, sulfonyl esters, aldehydes, glyoxal, epoxides, carbonates, aryl halides, imidoesters, carbodiimides, anhydrides, fluorophenyl esters, etc. Reactive group aldehydes or NHS esters are usually used. Reduced cysteine ​​(free sulfhydryl) can also be used. The method of combining payload molecules with antibodies is known. For example, the combination of payload molecules with antibodies is described in U.S. Patent No. 10,071,170, the description of which is incorporated herein by reference.

[0048] Examples of payload molecules include microtubule formation inhibitors, meiosis inhibitors, topoisomerase inhibitors, RNA polymerase inhibitors, DNA intercalators or alkylating agents, ribosomal inhibitors, siRNA, enzymes (carboxypeptidases, alkaline phosphatases, cytosine deaminases), immunocytokines (e.g., interleukin-2), and the like. Examples of cytotoxic drugs include, but are not limited to, maytansinoids, auristatins, dolastatins, tubulins, camptothecins, pyrrolobenzodiazepines, calicheamicin, baishunin, doxorubicin, polyamicin, carboplatin, cisplatin, cyclophosphamide, ifosfamide, nedran, bleomycin, mitomycin C, cytarabine, fluorouracil, methotrexate, trimetrexate, vinblastine, aritalin, hexamethylmelamine, procarbazine, paclitaxel, taxotere, diphtheria toxin, Pseudomonas exotoxin and its derivatives (e.g., PE38, PE40), alpha emitters (Ac-225, At-211, Th-227, Ra-223, Pb-212, Bi-212, Ra-224). Compounds include stereoisomers and derivatives thereof. In embodiments, the presently provided antibodies or antigen-binding fragments thereof specifically bind to an auristatin, which may be monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF).

[0049] In various embodiments, an ADC comprising a conjugate of an antibody and a cytotoxic agent can comprise any antibody or fragment or modification thereof that can be used to target a tumor antigen or to deliver a cytotoxic drug to a tumor cell. For example, several monoclonal antibodies have proven to be successful therapeutic agents for treating human cancers. These include rituximab, trastuzumab, cetuximab, panitumumab, bevacizumab, and many others. Examples of monoclonal antibodies against solid tumors include pertuzumab, ramucirumab, nivolumab, pembrolizumab, necitumumab, dinutuximab, oportuzumab, atezolizumab, avelumab, cemiplimab, carotuximab, margetuximab, bemarituzumab, naxitamab, relatlimab, brentuximab, lomvotuzumab, glembatumumab, BCD-100, spartalizumab, IBI308, CS1001, tremelimumab, TSR-042). Any of these antibodies can be used to prepare an antibody-drug conjugate.

[0050] Examples of antibodies and ADCs relevant to the present disclosure include, but are not limited to, gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine (TDM1), inotuzumab ozogamicin, polatuzumab vedotin, trastuzumab deruxtecan, trastuzumab duocarmazine, sacituzumab govitecan, loncastuximab tesirine, oportuzumab monatox, zolbetuximabclaudiximab, depatuxizumab mafodotin, mirvetuximab sorvatansine, rovalpituzumabtesirine, enfortumab vedotin, ladiratuzumab vedotin, zilovertamab vedotin, tisotumab vedotin, disitamab vedotin, BAT8001, L19IL2, and L19TNF.

[0051] In an embodiment, when the PBA is an antibody, the anti-drug antibody may be directed against the toxin (also referred to herein as a drug or payload) portion of the ADC. In an embodiment, the anti-drug antibody may be a fragment of a whole antibody. The fragment of the anti-drug antibody may be Fab, Fab', F(ab')2, Fv, scFv, single domain antibody or double antibody, or any other epitope binding fragment. The antibody fragment may be in the range of 0.5kDa-110kDa (Note: F(ab')2 is ~100kDa), including all Da values ​​and ranges therebetween. In one embodiment, the antibody fragment is about 15kDa. In one embodiment, the antibody is a single domain antibody (nanobody) containing only VHH (usually 13kDa to 15kDa). The anti-drug antibody has a binding affinity for the drug, which can be expressed by a dissociation constant (KD). In one embodiment, the KD of the anti-drug antibody is 1pM to 50nM, including all 0.1pM values ​​and ranges therebetween. In one embodiment, the KD is less than 1nM. In one embodiment, the KD is 1 pM to 100 pM. In one embodiment, the KD can be 1-100 pM. In one embodiment, the anti-drug antibody is a camelid, chimeric or humanized single domain antibody - also known as a nanobody. These antibodies have many properties that make them very suitable for current competitive inhibition methods. Single domain antibodies are a small antibody format (~15 kDa) that is very stable, can be expressed in E. coli, and can be humanized to limit immunogenicity. In addition, camelid immunization and phage display technology can quickly and cheaply develop new inhibitors.

[0052] Antibodies (including fragments or modifications) can be combined with free payloads or payloads cut from ADC. When the payload is cut from the ADC, for example, by enzymatic action, hydrolysis, oxidation or some other mechanism, the cut payload may include part of the linking group, all of the linking groups or no linking groups. The cut payload may also include a displacement group that can be added during or after the cut, or a functional group formed due to the cut process. Examples of displacement groups and / or functional groups formed by the cut process include, but are not limited to, alkyl groups (e.g., methyl, ethyl, propyl, isopropyl, etc.), alcohol groups, amines, thiols, sulfonic acids, sulfoxides, sulfides, sulfones, carboxylic acids, esters, amides, etc., and combinations thereof.

[0053] Various antibodies (including fragments or modifications) are described in the embodiments herein, and sequences are disclosed. The disclosure includes full-length antibodies, scFv, Fab and other antibodies specific for auristatin, including derivatives. The generation of antibodies, screening of antibodies to identify candidates with desired binding specificity and affinity, and their binding and inhibition properties are described.

[0054] The sequence of any antibody or fragment described herein may contain a polyhistidine tag or a linker, and sequences without polyhistidine tags may be used (e.g., to reduce immunogenicity), and when a linker is used, the binding construct is not limited to a specific linker. Any specific sequence disclosed herein with a polyhistidine tag also includes the corresponding sequence without a polyhistidine tag, and any sequence disclosed herein without a polyhistidine tag also includes a sequence with a polyhistidine tag. Variants of the antibody or fragment sequences disclosed herein include sequences with at least 85% identity to the disclosed sequence, as long as the binding affinity is not adversely affected. For example, the binding affinity of the variant may be 10% lower, the same or better than that of the disclosed sequence. In an embodiment, the variant may have at least 90%, at least 95%, at least 98%, at least 99% homology (identity) with the disclosed sequence without adversely affecting the binding affinity. The present disclosure also includes nucleotide sequences encoding amino acid sequences, or variants thereof, as described herein.

[0055] The present disclosure includes all polynucleotides encoding the antibodies and antigen-binding fragments thereof. Such polynucleotides include expression vectors comprising the encoding polynucleotides. The present disclosure includes cell cultures comprising expression vectors, and methods for producing the antibodies and antigen-binding fragments thereof by using cell cultures to produce antibodies or antigen-binding fragments and separating antibodies or antigen-binding fragments from cell cultures. Bacterial and mammalian cell cultures are included. The present disclosure also includes all polynucleotides that can hybridize with the polynucleotides encoding the antibodies or antigen-binding fragments. The hybridization can be such that the hybridization occurs in a solution at a temperature of about 20 degrees Celsius. The solution can contain a salt, such as a sodium salt, and its concentration can be about 0.15M.

[0056] PBAs can be peptides consisting of ten or more amino acids and have an equilibrium dissociation constant (K D ) is 50.0 nM or lower.

[0057] Surprisingly, the binder (e.g., MMAE) for the payload (drug) portion of the ADC does not adversely affect its efficacy in terms of cytotoxicity to cancer cells, but can reduce the toxicity associated with the unbound payload. Although not intended to be bound by any particular theory, it is possible that the entry and intracellular processing of the ADC may not be affected by the binding to the payload binder, and the cell entry of the unconjugated payload may be affected (reduced) by the binding to the payload binder. However, relative to the anticancer cell toxicity of the ADC, the payload binder can reduce the toxicity of the unconjugated payload, thereby enhancing the anticancer selectivity, which is unexpected. In one embodiment, when used in combination with a PBA (e.g., an antibody), the efficacy of the ADC may increase, in which case not only the non-target toxicity is reduced, but also the ADC efficacy is unexpectedly enhanced. In some embodiments, when the PBA is part of the ADC, it may not bind to the drug. Instead, it may only bind to the free drug. In this case, it is expected that there will be little or no effect on the efficacy of the ADC, but the non-target toxicity will be significantly reduced.

[0058] The present disclosure provides a composition for reducing off-target toxicity, which includes a means for reducing the toxicity of an unconjugated or cleaved payload (e.g., a drug) from an ADC without adversely affecting the efficacy of the ADC in the intended treatment (e.g., cancer cell treatment). The means for reducing off-target toxicity include anti-drug antibodies and fragments and modifications thereof, peptides, aptamers, Spiegelmers, fibronectin, DARPin, cyclodextrins and / or afitin.

[0059] In a non-limiting embodiment, a binding agent of the present disclosure specifically binds MMAE and comprises a light chain and a heavy chain comprising the following sequence:

[0060] ABC3319 sequence

[0061] Light chain (H55Y mutation in bold)

[0062]

[0063] Heavy chain (F27L mutation in bold, albumin binding sequence underlined)

[0064]

[0065] In one embodiment, ABC3319 (or any other binding partner of the present disclosure) may be adapted to bind to a substance in the blood of an individual, such as albumin or a blood cell surface protein, to increase its half-life. A non-limiting example of such a binding partner that binds to human serum albumin (which is an example of a substance in an individual) (and also specifically binds to MMAE) is referred to herein as ABC3320, wherein the light and heavy chains comprise the following sequences:

[0066] Light chain (H55Y mutation in bold)

[0067]

[0068] Heavy chain (F27L mutation in bold)

[0069]

[0070] In this example, the human serum albumin binding domain is shown in bold italics. The optional linker sequence can be replaced with any other suitable linker sequence, shown in italics. In an embodiment, a suitable linker can be 3-20 amino acids and can contain G, S or a combination thereof.

[0071] In embodiments, any binding partner described herein, including but not limited to the binding partners referred to herein as ABC3319, ABC3320, and ABC3315 (containing amino acid residues at the unchanged positions shown in Table 1 (i.e., in L_H55Y, H is the unchanged position and Y is the position where it is changed in the light chain)) can contain any one or combination of the amino acid substitutions (i.e., mutations) shown in Table 1. As described above, the positions of the point mutants shown in Table 1 are applicable to the sequences of any binding partner light and heavy chains having the amino acids at the same positions as in the ABC3315 binding partner. In Table 1, the prefix "L_" indicates a light chain and the prefix "H_" indicates a heavy chain. The same applies to all other amino acid changes described herein.

[0072]

[0073] Another amino acid change that can be used alone or in combination with the amino acid changes described in Table 1 includes the I101F change in the heavy chain. Therefore, the present disclosure explicitly includes all binding partners comprising one or more amino acid substitutions, as well as all sequences comprising or consisting of any one or any combination of said substitutions. In a non-limiting embodiment, the binding partner of the present disclosure comprises only Figure 2 , 4and 12. Thus, the present disclosure explicitly includes all binding partners comprising one or more amino acid substitutions, and all sequences comprising or consisting of any one or any combination of said substitutions. In addition, the present disclosure explicitly includes each binding partner comprising Figure 2 , Figure 4 , Fig.12 and any one or any combination of amino acid changes shown in Table 1. However, as described herein, the VH_F37V clone showed a significantly reduced binding signal relative to ABC3315. Therefore, in an embodiment, any binding partner described herein may exclude the mutation of F37 in its heavy chain.

[0074] As described above, in embodiments, PBAs can be engineered to bind to formed elements in the blood (e.g., albumin, erythrocyte membrane proteins, etc.). PBAs that have been engineered to bind to formed elements in the blood can be expected to exhibit reduced clearance (i.e., making administration more convenient) and increased blood: tumor exposure, thereby achieving greater pharmacokinetic selectivity (i.e., enabling a greater degree of inhibition of the release of payload molecules in the blood relative to inhibition of the release of payload molecules in the tumor). In embodiments, the components of the binder comprise an amino acid sequence that binds to albumin, further representative and non-limiting examples of which include the following sequences: QRLIEDICLPRWGCLWEDDF (SEQ ID NO: 6); QRLMEDICLPRWGCLWEDD (SEQ ID NO: 7); QRLMEDICLPRWGCLWE (SEQ ID NO: 8); and DICLPRWGCL (SEQ ID NO: 9).

[0075] In embodiments, the binding partners described herein are adapted to bind to erythrocytes. This adaptability may occur in the form of another binding partner that becomes a component of the anti-payload agent. In embodiments, the binding partners of the present disclosure thus comprise an amino acid sequence that confers the ability of the binding partner to bind to erythrocytes. In embodiments, the erythrocyte binding component comprises a single domain antibody.

[0076] In one aspect, the present disclosure provides a pharmaceutical composition comprising or consisting essentially of an ADC and an anti-payload agent as described herein. The formulation typically comprises a physiologically acceptable carrier, excipient or stabilizer, and may be in the form of an aqueous solution, lyophilized or other dry or solid formulation. Examples of suitable pharmaceutical formulation components can be found in Remington: The Science and Practice of Pharmacy 22th edition (2012). Acceptable carriers, excipients or stabilizers are non-toxic to the recipient at the doses and concentrations employed, including buffers such as phosphates, citrates, histidine and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzyl ammonium chloride; hexamethylammonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens, such as methyl or propyl parabens; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues); yl) polypeptides; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrins; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions, such as sodium; metal complexes (such as Zn-protein complexes); and / or nonionic surfactants, such as TWEEN TM 、PLURONICS TM , polyethylene glycol (PEG), etc. In one embodiment, the pharmaceutical composition may include a buffer component and a stabilizer, including but not limited to sucrose, polysorbate 20, NaCl, KCl, sodium acetate, sodium phosphate, arginine, lysine, trehalose, glycerol, and maltose. In an embodiment, the ADC and the anti-payload antibody or its fragment or modification are the only protein molecules present in the composition. In an embodiment, the ADC and the anti-payload antibody or its fragment or modification are the only antibodies present in the composition.

[0077] Compositions comprising ADC and payload binders can be administered together, or individually and independently using any suitable route, including parenteral, subcutaneous, intraperitoneal, intrapulmonary and intranasal. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. Administration can be performed in a continuous manner or intermittently. The appropriate dose will depend on the specific tumor being treated, the specific circumstances and conditions of the individual patient, the mode of administration, etc. The determination of the appropriate dose is within the purview of those skilled in the art (e.g., treating physicians). In one embodiment, ADC can be delivered locally, while PBA can be delivered so that it is available throughout the body. For example, ADC can be delivered at or near the tumor site or delivered intraperitoneally, while PBA can be delivered intravenously. In one embodiment, ADC can be delivered systemically, while anti-payload antibodies are delivered so that they are available throughout the body. For example, ADC can be delivered intravenously, while anti-payload antibodies can be delivered subcutaneously.

[0078] ADC and PBA can be administered as a single composition or as separate compositions. When administered as separate compositions, they can be administered sequentially or simultaneously. The two compositions can be administered at the same or different times, by the same or different routes, for the same or different lengths of time, and according to the same or different regimens.

[0079] In one embodiment, the amount of ADC and PBA alone or together is sufficient to reduce the non-target toxicity of ADC by at least 5% relative to the expected toxicity of the same concentration of ADC used alone and in combination. In an embodiment, the reduction in toxicity can be 10%, 20%, 30%, 40%, 50% or more. The reduction in non-target toxicity can be assessed by methods known in the art. For example, the reduction can be classified as a reduction in the percentage of patients experiencing grade 3 or higher adverse reactions at a specific dose (regardless of whether PBA is used) (Clin. Invest. (2013) 3 (12), 1157–1165). In addition, in a single patient, the reduction in non-target toxicity can be classified as a reduction in the severity of adverse reactions (e.g., neutropenia) with or without PBA.

[0080] ADC and PBA can be administered to individuals in need of treatment at doses effective for treating solid tumors. Typically, the suitable dosage range of PBA and ADC may be about 0.1 mg / kg to 100 mg / kg, including all 0.1 mg / kg values ​​and the range therebetween. Examples of dosages include 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 and 100 mg / kg. A variety of dosage regimens are contemplated, including dosage regimens (e.g., maintenance therapy) in which ADC and PBA can be repeatedly administered (e.g., on a daily, weekly or monthly schedule) in a short time or extended time (e.g., months to years). The application range of PBA may be 0.01–100 mg / kg, including all 0.01 mg / kg values ​​and the range therebetween. One skilled in the art can determine the appropriate ratio of ADC to PBA. For example, the molar ratio may be 1:1 to 1:100 ADC:PBA.

[0081] The amount of ADC and anti-payload antibody can be administered to individuals in need of treatment at a dosage effective for treating solid tumors. Typically, the appropriate dosage of an antibody or its fragment (for ADC and anti-payload antibody) can be about 0.1 mg / kg to 100 mg / kg, including all 0.1 mg / kg values ​​and the range therebetween. Examples of dosages include 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 and 100 mg / kg. A variety of dosing regimens are contemplated, including dosing regimens that can be repeatedly administered with ADC and anti-payload antibodies, for example, on a daily, weekly or monthly schedule, in a short period of time or over an extended period of time, such as months to years (e.g., maintenance therapy). The application range of anti-payload antibodies can be 0.01-100 mg / kg, including all 0.01 mg / kg values ​​and the range therebetween. One skilled in the art can determine the appropriate ratio of ADC to anti-drug antibody. In one embodiment, the molar ratio can be 1:1 to 1:100 ADC:anti-drug antibody.

[0082] As described above, in embodiments, half-life extension strategies can be used to increase the plasma half-life of anti-payload antibodies, including fusion of moieties to sdAbs or Fabs that bind blood components (e.g., albumin, red blood cells (e.g., band 3 of RBCs), or endogenous IgGs) and PASylation and PEGylation. Thus, in embodiments, anti-payload antibodies can be fused to moieties that bind albumin, red blood cells, or endogenous IgGs or fragments thereof, or can be PASylated and / or PEGylated. For example, a bispecific antibody can have an arm that binds to a payload and another arm that binds to albumin or red blood cells (e.g., band 3 of RBCs). In embodiments, the antibody or fragment does not contain a polyhistidine tag. Therefore, it is believed that sdAb-sdAb fusion proteins that combine anti-payload binding activity and anti-albumin or anti-red blood cell binding activity may exhibit ideal pharmacokinetic properties (restricted distribution in tissues and tumors relative to distribution in the blood, elimination by renal filtration due to low molecular weight, and long half-life), and can provide optimal enhancement of ADC therapeutic selectivity (increasing the ratio of efficacy to off-site toxicity). It is also believed that sdAb-peptide or Fab-peptide fusion proteins that combine anti-payload binding affinity (via sdAb or Fab) and albumin binding affinity (via peptide) may exhibit desirable pharmacokinetic properties (restricted distribution within tissues and tumors relative to distribution within blood, elimination by renal filtration due to low molecular weight, long half-life) and may provide optimal enhancement of ADC therapeutic selectivity (increased ratio of efficacy to off-site toxicity).

[0083] Representative examples of single domain antibodies that have been demonstrated to bind to erythrocytes via band 3 are as follows:

[0084] >RMC1

[0085] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:10)

[0086] >RA8

[0087] QVQLVQSGGGLVQAGGSLRLSCAASERTFSTYAMGWFRQIPGKERLFVAAVNWNGKTIRYADSVKGRFTISRDNAKNTIALQMNSLKPEDTAVYYCALRSTPMYFTNLASQESYNYWGPGTQVTVSS(SEQ ID NO:11)

[0088] >RB12

[0089] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYEYRTYDYWGQGTQVTVSS(SEQ ID NO:12)

[0090] >RD1

[0091] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYEYRTYDYWGQGTQVTVSS(SEQ ID NO:13)

[0092] >RD11

[0093] EVQLVESGGGLVQPGGSLRLSCAASGRIFSISNMGWYRQAPGKQRELVATITSGGSTNYGDSVKGRFTISMANAKNAVYLQMNSLKPEDTAVYYCNAGISRRTGTYSGGRYSDYWAQGTQVTVSS(SEQ ID NO:14)

[0094] >RE8

[0095] QVQLVQSGGGLVQPGGSLRLSCAASGRVSEINTMGWYRQAPGKQRELVALITSTSSTMYSDSVKGRFTVSRDVAKNMVYLQMNSLKPEDTAVYYCNARQTKWYAAGYEYRTYDYWGQGTQVTVSS(SEQ ID NO:15)

[0096] >RG10

[0097] QVQLQESGGGLVQAGGSLRLSCAASERTFSTYAMGWFRQTPGKERLFVAAVNWNGKTIRYADSVKGRFTISRDNAKNTMSLQMNSLKPEDTAVYYCALRSTPMYFTNLASQESYNYWGPGTQVTVSS(SEQ ID NO:16)

[0098] >RH5

[0099] QVQLVQSGGGLVQPGGSLRLSCAASGRIFSISNMGWYRQAPGKQRELVATITSGGSTNYGDSVKGRFTISMVNAKNAVYLQMNSLKPEDTAVYYCNAGISRRTGTYSGGRYSDYWGQGTQVTVSS(SEQ ID NO:17)

[0100] >RMA1

[0101] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYTDSVKGRFTISRDKAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYVYRTYDYWGQGTQVTVSS(SEQ ID NO:18)

[0102] >RMD1

[0103] QVQLVQSGGGLVQPGGSLRLGCAASGRVSEINTMGWYRQAPGKQRGLVALITSTSSTMYSDSVKGRFTVSRDVAKNMVYLQMNSLKPEDTAVYYCNARLTRWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:19)

[0104] >RMF1

[0105] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVNGRFTISRDNAKNTVYLQMTSLEPEDTAVYYCHARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:20)

[0106] >RMG1

[0107] QVQLVQSGGGLVQPGGSLRLSCAASGRVSEINTMGWYRLAPGKQRELVALITSTSSTMYSDSVKGRFTVSRDVAKNMVYLQMNSLKPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:21)

[0108] >RMC2

[0109] QVQLVQSGGGLVQPGGSLRLSCAASGRVSEINTMGWYRQAPGKQRELVALITSTSSTMYSDSVKGRFTVSRDVAKNMVYLQMNSLKPEDTAVYYCNARQTIWYAAGYKYRTYDYWGLGTQVTVSS(SEQ ID NO:22)

[0110] >RME2

[0111] QVQLVQSGGGLVQSGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYEYRTYDYWGQGTQVTVSS(SEQ ID NO:23)

[0112] >RMF2

[0113] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNNDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:24)

[0114] >RMG2

[0115] QVQLVQSGGGQVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:25)

[0116] >RMH2

[0117] QVQLVQSGGGLAQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:26)

[0118] >RMB3

[0119] QVQLVQSGGGLVQPGGSLRLSCAASGRVSEINTMGWFRQAPGKQRELVALITSTSSTMYSDSVKGRFTVSRDVAKNMVYLQMNSLKPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:27)

[0120] >RMD3

[0121] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLQMASLEPEDTAVYYCNARQTRWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:28)

[0122] >RME3

[0123] QVQLVQSGGGLVQPGGSLRLSCAASGRVSEINTVGWYRQAPGKQRELVALFTSTSSTMYSDSVKGRFTVSRDVAKNMVYLQMNSLRPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:29)

[0124] >RMG3

[0125] QVQLVQSGGGLVQAGGSLRLSCAASGSSSSIIAMGWYRQAPGKQRELVATISNGDTTNYIDSVKGRFTISRDNAKNTVYLRMASLEPEDTAVYYCNARQTKWYAAGYKYRTYDYWGQGTQVTVSS(SEQ ID NO:30)

[0126] The compositions of the invention may be administered alone or in combination with other types of treatment, such as surgical resection, radiation therapy, chemotherapy, hormone therapy, immunotherapy or other anti-tumor agents.

[0127] The compositions of the present invention can be used for any type of cancer, including carcinoma, lymphoma, sarcoma, melanoma and leukemia. Non-limiting examples include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, peritoneal cancer, myeloma (including multiple myeloma), hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, glioblastoma / glioma (e.g., anaplastic astrocytoma, glioblastoma multiforme, anaplastic oligodendroglioma, anaplastic oligoastrocytoma), cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, brain cancer, colon cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer, liver cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer and various types of head and neck cancer.

[0128] In one aspect, the present disclosure provides a method for generating and identifying anti-payload antibodies suitable for use in combination with ADC therapy to reduce non-target cytotoxicity. The method comprises generating or obtaining an antibody library, identifying specific antibodies based on positive binding to the payload molecule by ELISA and / or surface plasmon resonance technology, identifying antibodies with a desired affinity (e.g., K D The invention relates to antibodies with a binding affinity of 50 nM or less, determining binding kinetic parameters, and determining in vivo efficacy. Any auristatin described herein may be a monomethyl auristatin.

[0129] In one embodiment, the present disclosure provides compositions and methods for treating cancer using ADC, wherein the drug portion of the ADC is auristatin E (MMAE) or auristatin E derivative, analog or metabolite, or auristatin F, or auristatin F derivative, analog or metabolite, or a related auristatin derivative. The method includes administering an ADC to an individual in need of treatment, wherein the drug portion is the aforementioned auristatin, and a PBA directed to the auristatin portion of the ADC. The ADC and the PBA can be administered in the same composition or in separate compositions as described elsewhere in the present disclosure. An example of an ADC whose drug portion is auristatin E is brentuximab vedotin. Another example of an ADC whose drug portion is auristatin E is polatuzumab vedotin. When the drug portion of the ADC is auristatin E, examples of PBAs that can be used are IgM D9, sdAb MA3, MB2, and MC7. Also provided is a method for treating cancer (eg, solid tumors), comprising administering an ADC to an individual in need of treatment, wherein the drug is auristatin E and the antibody, which may be IgM D9, sdAb MA3, MB2, and MC7.

[0130] When the drug moiety of the ADC is an auristatin, examples of PBAs that can be used are described herein and in the figures. In an embodiment, the PBA is any one of ABC3320, ABC3319, ABC3317, ABC3315, 1B3, 1H2, 3B8, 2E8, or 2C2, or a combination thereof.

[0131] In a non-limiting embodiment, the present disclosure is used in conjunction with an ADC that is Polatuzumab Vedotin (PV).

[0132] Non-limiting examples of some PBA sequences provided by the present disclosure are as follows.

[0133] Cloned sequences identified from immunolibrary screening

[0134] 1B3

[0135] DIVMTQSHKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKIEESGGGLVKAGGSLKLSCAASGFTFSRYDMSWVRQTPEKRLEWVATISSGGRHTYYPDSVKGRFTISRDNVKNTLYLQMNSLRSVDTAMFYCLASMLTTDYFEYWGQGTSLTVSS (SEQ ID NO:31)

[0136] 1H2

[0137] DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCQQYNSYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL (SEQ ID NO:32)

[0138] 3B8

[0139] DVVMTQTQKFMSTSVGDRVSITCKASQNVGTAVAWYQQKPGQSPKLLIYSASNRYTGVPDRFTGSGSGTDFTLTISNMQSEDLADYFCQQYSSYPYTSGGGTKLEIKRGGGGSGGGGSGGGGSSGGGSQVQLVETGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQTPEKRLEWVATISSGGSYTYYPDSVKGRFTSSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVL(SEQ ID NO:33)

[0140] Sequence m in the mutagenesis library = mutagenesis, A / B = plate, 3 / 24 = dissociation time, letter # = well (see Example)

[0141] [1]MA24G1

[0142] [2]MDIVMTQSHKFMSTSVGDRVSITCKASQDVDTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:34)

[0143] [3]MA24E2

[0144] [4]MDVVMTQTQKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRCGGGSGGGGSGGGGSGGGGSQVQLVETGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:35)

[0145] [5]MA24C5

[0146] [6]MDVVMTQTQKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPVRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLVETGGGLVKPGGSLKLSCAASSSTFSGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:36)

[0147] [7]MA24B5

[0148] [8]MDIVMTQSHKFMSTSVGDRVSITCKASQDVDTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:37)

[0149] [9]MA24H7

[0150]

[10] MDIVMTQSQKFMSTSVGDRASVTCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISEVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:38)

[0151]

[11] MA24G6

[0152]

[12] MDIVMTQSHKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTPHTGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCQQYNSYPYTFGGGTKLEIKRGGGGSGGGGSGGGDSGGGGSEVKLVESGGGLVIPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDIAKNTLYLQMSSLRSEDTAMYYCLASLFTTDYFEYWGQGTTVTVPL(SEQ ID NO:39)

[0153]

[13] MA24B10

[0154]

[14] MDIVMTQSHKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAARGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:40)

[0155]

[15] MA24B2

[0156]

[16] MDIVMTQSQKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLQIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPENRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:41)

[0157]

[17] MB24E2

[0158]

[18] MDIVMTQSHKFMSTSIGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLVETGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:42)

[0159]

[19] MB24D9

[0160]

[20] MDIVMTQSHKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTLNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRNEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:43)

[0161]

[21] MB24E4

[0162]

[22] MDIVMTQSHKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKIEESGGGLVKAGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:44)

[0163]

[23] MB24E3

[0164]

[24] MDVVMTQTQKFMSTSVGDRVSITCKASQNVGTAVAWYQQKPGQSPKLLIYSASNRYTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPENRLEWVATISSGGSYTYYPDSVKGRFTSSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:45)

[0165] MB24D10

[0166] MDIVMTQSHKFVSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTLHTGVPDRFTGSGSGTDFTLTISNVQNEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYSEYWGQGTTVTVLP(SEQ ID NO:46)

[0167] MA3B10

[0168] MDVVMTQTQKFMSTSVGDRVSITCKASQNVGTAVAWYQQKPGQSPKLLIYSASNRYTGVPDRYTGSGSGTDFTLTISNMQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTFYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASMLTTDYFEYWGQGTTVTVLL(SEQ ID NO:47)

[0169] MA3B11

[0170] MDIVMTQSHKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSGGGGSEVKLVESGGGLVKPGGSLKLSCAASGFTFNRYALSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:48)

[0171] MB3G6

[0172] MDIVMTQSHKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLVETGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQTPEKRLEWVATISSGGSYSYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYYCLASLITTDYFEYWGQGTTVTVLL(SEQ ID NO:49)

[0173] MB3E8

[0174] MDVVMTQTQKFMSTSVGDRVSITCKASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSQVQLVETGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNAKNTLYLQMSSLRSEDTAKYYCLASLFTTDYFEYWGQGTTVTVLL(SEQ ID NO:50)

[0175] ABC3315 humanized Fab sequence (predicted CDR sequences are shown in bold)

[0176] Light chain

[0177]

[0178] Heavy chain

[0179]

[0180] The present disclosure includes antibodies and antigen-binding fragments thereof having Figure 4 All amino acid substitutions (individually and in all combinations) are shown, including but not limited to those specified within the boxes.

[0181] In one aspect, the present disclosure provides a kit comprising an ADC and a component for reducing the non-target toxicity of the ADC. The kit may comprise the same or different compositions, i) ADC, and ii) an anti-payload antibody, wherein the anti-payload antibody is specific to the drug portion of the ADC. The ADC and the anti-payload antibody may be provided in powdered, lyophilized form, together with a reconstitution medium, wherein the antibody and ADC may be reconstituted prior to use. The kit may also optionally include instructions for administering a composition comprising an ADC and a composition comprising an anti-payload antibody, which compositions may be administered by different routes.

[0182] The following paragraphs provide some non-limiting embodiments.

[0183] Embodiment 1. A method of reducing off-target toxicity of an antibody-drug conjugate (ADC), comprising administering to an individual in need of treatment an ADC and a payload binding agent (PBA) directed to a drug moiety of the ADC. In various embodiments, the drug moiety is a payload cleaved from the ADC.

[0184] Embodiment 1a. The method of embodiment 1, wherein the PBA is an antibody or fragment or modification thereof directed against the drug portion of the ADC.

[0185] Embodiment 1b. The method of embodiment 1, wherein the PBA is a peptide.

[0186] Embodiment 2. The method of embodiment 1, wherein the ADC and the PBA are administered in the same composition.

[0187] Embodiment 3. The method of embodiment 1, wherein the ADC and the PBA are administered in different compositions.

[0188] Embodiment 4. The method of embodiment 3, wherein the ADC and the PBA are administered by different routes.

[0189] Embodiment 4a. The method of embodiment 4, wherein the ADC is administered by an intraperitoneal route and the PBA is administered by an intravenous route.

[0190] Embodiment 5. The method of embodiment 1, wherein the drug is auristatin E or auristatin F.

[0191] Embodiment 6. The method of embodiment 1, wherein the K of PBA D Less than or equal to 50nM.

[0192] Embodiment 6a. The method of embodiment 1, wherein the K of the anti-drug antibody is D Less than or equal to 50nM.

[0193] Embodiment 7. The method of embodiment 6 or 6a, wherein the K of the PBA (embodiment 6) or the anti-drug antibody (embodiment 6a) is D Less than 1nM.

[0194] Embodiment 8. The method of embodiment 7, wherein the K of the PBA or anti-drug antibody D 1pM to 100pM.

[0195] Embodiment 9. A composition comprising an ADC and a PBA directed against the drug portion of the ADC.

[0196] Embodiment 9a. The composition of embodiment 9, wherein the PBA is an antibody or a fragment or modification thereof directed against the drug portion of an ADC (anti-drug antibody).

[0197] Embodiment 10. A kit comprising: i) a composition comprising an ADC; ii) a composition comprising an anti-drug antibody specific for a drug in the ADC; iii) optionally, instructions for use, including instructions for administration of i) and ii).

[0198] The invention is further illustrated by the figures and data provided herein, whether provided in the detailed description or in the figures.

[0199] Example 1

[0200] This example provides a description of the compositions and methods used in the present disclosure, particularly for identifying mouse antibodies against MMAE.

[0201] method

[0202] immunity

[0203] Monomethyl auristatin F (MMAF) is conjugated to Aplysia hemocyanin (KLH) or BSA via an EDC linker. Briefly, 0.5 mg MMAF was dissolved in MES buffer (0.1 M MES, pH 4.7) / 30% DMF and mixed with 2 mg KLH or BSA in MES buffer, followed by the addition of 0.5 mg EDC dissolved in water. The solution was incubated overnight at room temperature. Single animal immunization was performed using approximately 50 μg KLH-MMAF immunogen emulsified in Freund's incomplete adjuvant. Female Balb / c mice were subcutaneously injected with 200 μL of emulsion and boosted every three weeks. Anti-MMAF-BSA plasma titers were assessed by ELISA using an anti-mouse Fc secondary antibody coupled to alkaline phosphatase.

[0204] RNA Isolation

[0205] The spleen was surgically removed from the immunized mice and separated using forceps and a needle. The released splenocytes were collected in a 15 ml conical tube. Lysis and homogenization of the lymphocyte sample in 5 ml TRIzol reagent followed by incubation for 5 min and the addition of 1 ml chloroform. The mixture was vortexed for 30 s, incubated for 3 min, and centrifuged at 12,000 × g at 4°C for 15 min. The mixture separated into 3 layers: a lower red phenol-chloroform layer, an intermediate layer, and a colorless upper aqueous phase. The aqueous phase containing RNA was transferred to a new tube, 2.5 ml isopropanol was added, and incubated for 10 min. Total RNA was precipitated as a white gelatinous precipitate by centrifugation at 12,000 × g at 4°C for 10 min. The supernatant was discarded and the pellet was suspended in 5 ml 75% ethanol. The sample was briefly vortexed and then centrifuged at 10,000 × g at 4°C for 5 min. The supernatant was discarded and the pellet was air-dried for 15 min. Resuspend the pellet in 200 μL of RNAse-free water and incubate at 60 °C for 10 min to ensure that the total RNA is completely dissolved. Store the RNA samples at -80 °C for subsequent processing.

[0206] cDNA synthesis

[0207] Mix the following reagents, heat at 65°C for 5 minutes, and then incubate on ice for at least 1 minute: 1 μl 50 μM Oligo d(T)20 primer, 1 μl 10 mM dNTP mix (10 mM each), 2.5 μl template RNA, and 9.5 μl nuclease-free water. Add the following reagents to the RNA-primer mixture: 4 μl 5× SSIV buffer, 1 μl 100 mM DTT, 1 μl RNaseOUT TM Recombinant RNase inhibitor and 1 μl IV reverse transcriptase (200 U / μL). The combined reaction mixture was incubated at 50-55°C for 15 minutes and then inactivated at 80°C for 10 minutes.

[0208] DNA amplification and scFv

[0209] Mouse variable heavy (VH) and light (VL) chains were amplified by polymerase chain reaction (PCR) using the following reagents: 25 μl OneTaqGC 2X master mix, 2 μl cDNA, 0.5 μl forward primer, 0.5 μl reverse primer, and 22 μl nuclease-free water. PCR products were obtained after 30 cycles: denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 68°C for 45 seconds, and final extension for 5 minutes. Amplified VH and VL products (~400 bp) were size-selected and purified on 1% agarose gel, and then mouse scFv with added SfiI cloning sites was assembled by overlap extension PCR, with the following reagents: 25 μl OneTaqGC 2X master mix, 10 ng each of purified VH and VL, 1 μl each of forward and reverse primers, 5 μl GC Enhancer, and 50 μl of water. PCR conditions were as follows: 95°C denaturation for 1 minute, 63°C annealing for 1 minute, 72°C extension for 1 minute, 5 cycles; then 95°C denaturation for 1 minute, 56°C annealing for 30 seconds, 72°C extension for 1 minute, 5 cycles; followed by 25 cycles: 95°C denaturation for 1 minute, 72°C annealing and extension for 90 seconds, and a final extension for 5 minutes. The amplified scFv product (~850 bp) was size selected and purified on a 1% agarose gel.

[0210] Construction of immune phage display library

[0211] For test library construction, the amplified PCR product (1 μg / replicate) and pADL-10b phagemid vector (2 μg / replicate) were digested three times with SfiI restriction enzyme according to the manufacturer's recommendations. The reaction was incubated at 50°C for 16 hours. The cleavage products were recovered by DNA gel electrophoresis using 1% agarose gel. The test library was first developed by ligating the digested DNA into the phagemid vector. The ligated vector was purified with the EZNA Cycle Pure Kit. Electroporation transformed electrocompetent TG-1 cells using 2 μL DNA with the following conditions: 1.0 mm cuvette, 10 μF, 600 Ohms, 1800 V, time constant 3.5 to 4.5 milliseconds. Immediately after pulsing the cells, 975 μL recovery medium was added to the cells. The cell suspension was then transferred to a 3 mL tube and incubated at 37°C at 250 rpm for 1 hour. The cell suspension was then serially diluted to 1:1000 on LB agar plates containing 100 μg / ml ampicillin and 2% (wt / v) glucose and incubated overnight at 37°C. The electroporation efficiency was determined by counting colonies and multiplying by the corresponding dilution factor. Approximately 10-20 clones were isolated, resuspended in water, and incubated at 95°C for 5 minutes, followed by centrifugation at 4°C for 5 minutes. DNA in the supernatant was amplified as previously described using 5' and 3' sequencing primers. PCR products were purified and sequenced to assess the functionality and size of the library.

[0212] To construct the complete library, ligate enough DNA into pADL-10b to generate at least 10 7 Libraries of individual transformants. Perform several separate electroporations using 3 μL of ligated DNA and 25 μL of TG1 cells. All electroporations are combined into a 50 mL conical tube. Serially dilute the transformed bacteria to 1:10,000 on LB agar plates containing 100 μg / ml ampicillin and 2% (wt / v) glucose and grow overnight at 37°C. Estimate library size by colony counts. Perform fragment PCR and sequencing as described above. Inoculate the remaining bacterial suspension onto four 245 mm square LB agar plates containing 100 μg / ml ampicillin and 2% (wt / v) glucose and grow overnight at 37°C. Repeat this process until the library size is ≥10 7 The library was recovered by first scraping bacteria from a 245 mm petri dish and placing them into 8 mL of LB medium, which was then combined into a 50 ml conical flask. The collected cell suspension was mixed with sterile glycerol (20%, v / v). Cell library aliquots were stored at -80°C.

[0213] The thawed library aliquot was inoculated into 60 mL of 2xYT medium supplemented with 100 μg / mL ampicillin and 2% (wt / v) glucose in a baffled 250 mL Erlenmeyer flask. The inoculated culture was grown at 37°C until OD600≈0.5. To rescue phage, 10 mL of the culture was transferred to a 50 mL Erlenmeyer flask, 1 μL of CM13 helper phage was added, and incubated at 37°C at 250 rpm for 1 hour. The infected cells were then separated by centrifugation at 2,800xg for 10 minutes at room temperature. The cells were suspended in 50 ml of 2×YT supplemented with 100 μg / mL ampicillin and 50 μg / mL kanamycin in a baffled 250 ml Erlenmeyer flask and incubated overnight at 30°C and 200 rpm. Transfer the bacterial culture to two 50 ml conical tubes and centrifuge at 3,200 x g and 4 ° C for 15 minutes to pellet the cells. Separate the supernatant into a new 50 ml conical tube, add 6 ml of 20% (wt / v) PEG6000 / 2.5M NaCl solution to it, invert to mix, and cool on ice for 30 minutes. Centrifuge at 3,200 x g and 4 ° C for 10 minutes to precipitate the phage particles. Resuspend the precipitated phage particles in 1 mL of ice-cold PBS and transfer to a microcentrifuge tube. Centrifuge the phage particle suspension at 16,000 x g and 4 ° C for 90 seconds to precipitate any residual bacteria. Reprecipitate the phage particles in the supernatant by adding 250 μl of ice-cold 20% (wt / v) PEG6000 / 2.5M NaCl solution, invert to mix, and incubate on ice for 10 minutes. The phage particles were centrifuged at 16,000xg and 4°C for 20 minutes and then resuspended in 0.5mL ice-cold PBS. The remaining bacterial debris was removed by centrifugation at 16,000xg and 4°C for 90 seconds. The phage concentration was determined by titration before panning. In brief, the phage was diluted 10 times in PBS. 10μL of each dilution was added to a single well of a low-binding 96-well round-bottom culture plate, each containing 90μL TG1 cells (pre-culture suspension). The culture plate was incubated at 37°C for 15 minutes to infect TG1 cells. Approximately 5μL of infected TG1 cells in each dilution were transferred to solid selective medium (LB+100μg / mL ampicillin+2% glucose) and grown overnight at 37°C. The phage concentration was determined by counting the colonies grown on the highest dilution culture plate.

[0214] Biopanning and screening of mouse scFv

[0215] MMAF was conjugated to biotin-PEG2-amine for biopanning and screening of anti-MMAE scFv. 0.5 mg MMAF was dissolved in MES buffer (0.1 M MES, pH 4.7) / 30% DMF and mixed with 100 μg biotin-PEG2-amine in MES buffer, followed by the addition of 1.5 mg EDC dissolved in water. The reaction was incubated overnight at room temperature and stored at 4 °C. The streptavidin beads were washed three times with PBST (PBS + 0.05% Tween 20) and blocked with MPBS (PBS + 5% skim milk powder) for 2 hours. After the blocking step, the streptavidin beads were incubated with 1 mL 1 μM biotin-MMAF for 15 minutes and then washed three times with PBST. For the first panning input, the phage stock was diluted to 10 12 cfu (colony forming units) / mL of blocking buffer (2% milk PBS), 1 ml of diluted phage was added to MMAF-biotin coated streptavidin beads and incubated for 2 hours. For subsequent panning, the original phage was diluted 1: 1 in blocking buffer. After incubation, the beads were washed 5 times, 10 times, 15 times and 15 times with PBST in the 1st, 2nd, 3rd and 4th rounds of panning. Then, in the 1st, 2nd, 3rd and 4th rounds of panning, the bound phage was incubated in PBS with free MMAE at concentrations of 1 μM, 100nM, 10nM and 1nM for more than 1 hour to elute. The output phage was titrated and reinfected with TG-1 cells to produce phage for the next round of panning.

[0216] After the fourth round of panning, phage-infected TG1 cells were grown overnight, serially diluted in 2xYT medium, and spread on individual plates containing selective medium (LB agar + 100 μg / mL ampicillin + 2% wt / v glucose) and incubated overnight at 37°C. Master plates were generated by inoculating single colonies into wells of a 96-well round-bottom culture plate containing 100 μL 2xTY (supplemented with 100 μg / mL ampicillin, 2% (wt / vol) glucose, and 15% (vol / vol) glycerol) and growing overnight at 37°C, 300 rpm. These wells were then used to inoculate wells of a 96-well deep-well plate containing 1 mL 2xTY medium (100 μg / mL ampicillin per well). The plate was incubated at 37°C and 300 rpm for 4 hours until OD600≈0.5, and then 1 μL of reserve helper phage was added to each well and incubated for another 1 hour. Kanamycin was added to a final concentration of 50 μg / mL and the plates were incubated overnight at 30°C, 250 rpm.

[0217] Nunc Maxisorp 96-well ELISA plates were coated overnight at 4 °C with 4 μg / ml NeutrAvidin. The plates were washed five times with PBST, blocked with MPBS for 2 h at room temperature, and 100 μL of 1 μM Biotin-MMAF was added for 30 min. The plates were then washed five times with PBST and incubated with 4-fold diluted phage supernatant for 2 h, with pre-incubation with free MMAE at concentrations of 10 nM and 100 nM, and 100 nM MMAE-ADC. The plates were then washed five times with PBST, and bound scFv-displaying phages were detected for 1 h using anti-M13 phage HRP-conjugated antibody (Antibody Design Labs, San Diego, CA) diluted 1:1000 in MPBST. After washing five times with PBST, 100 μL of 1-Step Turbo TMB-ELISA solution was added to each well and incubated for 15 min. The reaction was quenched by adding 100 μL of stop solution to each well, and absorbance was measured at 450 nm. DNA isolated from positive clones was sent to the Roswell Park sequencing core facility (Buffalo, NY) for sequencing.

[0218] mutagenized library

[0219] The genes of scFv clones 3B8, 1H2, and 1B3 were codon-optimized for Escherichia coli and synthesized by GeneArt. The heavy and light chains of each clone were amplified by PCR and subsequently mutated using a PCR-based random mutagenesis method. The purified mutagenized products were combined, and the heavy and light chains were ligated using the overlap extension PCR method. The complete library was constructed following the method for constructing an immunophage library, using the phagemid vector pComb3XSS instead of the pADL-10b phagemid vector.

[0220] Panning and screening of the mutagenized library

[0221] Phages were panned against MMAF-peg11-biotin-streptavidin-coated beads for three rounds with increasing stringency. After the third round of panning, phages displaying scFv were dissociated with 1 μM MMAE for 3 h, the supernatant was removed, and the remaining phages were eluted with 1 μM MMAE for 24 h. Single colonies from the 24-h elution were inoculated into two 96-deep-well plates and phages were expressed in deep-well plates following the above protocol. The next day, the supernatant containing phages was diluted 1:10 in 2% milk-PBS in individual wells of a 96-well plate with or without 1 nM free MMAE for 1 h. Subsequently, the solution containing phages was transferred to the wells of a Nunc Maxisorb plate containing immobilized MMAF-peg11-biotin-neutravidin and incubated in a shaking incubator at room temperature for 2 h. The plates were washed and bound phages were detected. Colonies with >70% signal knockdown were sent for DNA sequencing.

[0222] Humanization

[0223] The amino acid sequences of the variable heavy and variable light chains of clone MA24E2 were imported into Abysis. Murine framework residues that occurred less frequently in human antibodies were conservatively mutated to amino acids with higher frequency. The murine and humanized sequences were imported into Abodybuilder to create structural model predictions for the human and murine sequences. The predicted structures were superimposed in ChimeraX to ensure that the predicted human sequence had a similar structure to the murine sequence. The humanized sequences were expressed as fab fragments (ABC3315) in ExpiCHO-S cells and expressed using CaptureSelect according to the manufacturer's recommendations. TM The samples were purified by CH1-XL affinity resin.

[0224] Competitive ELISA

[0225] Anti-MMAE fab was diluted to 1nM and incubated with a series of concentrations of MMAE, MMAF, and trastuzumab-vc-MMAE. The solution was added in triplicate to each well of an ELISA plate fixed with MMAF-peg11-biotin-streptavidin. The plate was incubated for two hours at room temperature on a shaking platform set to 300rpm. The wells were washed 4 times with PBST, and 250μL of a 1:1,000 dilution of anti-human AP secondary antibody was added to each well and incubated for 1.5 hours. The wells were washed 2 times with PBST and 2 times with distilled water. 250μL of 4mg / ml PnPP (dissolved in diethanolamine) was added to each well, and the absorbance at 405nm was evaluated over time within 10 minutes. The absorbance changes over time of MMAE / MMAF and trastuzumab-vc-MMAE wells were normalized to the wells treated with anti-MMAE fab alone to determine the binding score of anti-MMAE fab.

[0226] Surface plasmon resonance

[0227] Binding of anti-MMAE fabs was assessed using SR7500DC surface plasmon resonance. MMAF-peg11-biotin was flowed through the left channel of a neutravidin-immobilized SPR chip. Unbound sites on both channels were subsequently blocked by injection of free biotin. ABC3315 fab was injected continuously for 3 minutes at concentrations of 1.23, 3.70, 11.11, 33.33, and 100 nM, with a 3-hour dissociation step after the last 100 nM injection. To assess binding of ABC3315 to free MMAE and PV, ABC3315 was conjugated to nhs-peg12-biotin and injected onto the left channel of a streptavidin SPR chip. Unbound sites on both channels were subsequently blocked by injection of free biotin. MMAE was injected continuously for 3 minutes at concentrations of 0.37, 1.11, 3.33, 10, and 30 nM, with a 3-hour dissociation step after the last 30 nM injection. A second kinetic titration was performed when PV was injected at bound MMAE concentrations of 1.23, 3.70, 11.11, 33.33, and 100 nM, with a 3-hour dissociation step after the final 100 nM injection. The observed sensorgrams were fitted using the Kinetic Titration Module of ClampXP to obtain the association rate constant, dissociation rate constant, and equilibrium dissociation rate constant.

[0228] Cell viability assay

[0229] Ramos cells at a density of 50,000 cells / mL were divided into individual wells of a 96-well flat-bottom culture plate. Culture medium containing MMAE or PV dilutions (with or without 500nM anti-MMAE fab) was added to each well, and the cells were incubated in a humidified incubator at 37°C (containing 5% CO2) for 4 days. On the fourth day, 25μL 4mg / ml MTT was added to each well, and the culture plate was incubated for 2 hours. Subsequently, MTT was dissolved overnight after adding 100μL 10% SDS, 0.1M HCL. The absorbance of each well was read at 550nm and 690nm wavelengths. A similar protocol was followed to evaluate the effect of anti-MMAE fab on the efficacy of trastuzumab-vc-MMAE and MMAE on SKBR3 cells. SKBR3 was trypsinized and diluted to a concentration of 40,000 cells / mL. 100 μL of cell suspension was added to each well of a 96-well U-bottom culture plate and cells were allowed to attach overnight. The next day, the medium was aspirated and replaced with fresh medium containing MMAE or trastuzumab-vc-MMAE with or without 500 nM anti-MMAE fab. Cells were incubated with fresh medium for 6 days, and drug dilutions were added on days 3 and 5. Cell viability was assessed using the same MTT protocol as for RAMOS cells. Cell viability was determined by dividing the difference in absorbance at 550 nm and 690 nm for treated wells by the difference in absorbance for untreated wells. The observed cell viability for each group was fitted to the four-parameter inhibitor-response equation in GraphPad Prism7.

[0230] Xenotransplantation efficacy study

[0231] Male and female Nu / J mice (Jackson Laboratories) were injected above the right hind leg with 5x10 6 Ramos cells. The growth of xenografts was monitored using a digital vernier caliper and measured with W 2 Tumor volume was calculated as xL / 2, where L is the longest diameter of the tumor and W is the diameter of the tumor perpendicular to L. When the tumor volume is 200-300mm 3 (Average ~250mm 3), mice were randomly divided into 8 groups to receive (i) PBS+PBS control, (ii) 1 mg / kg PV+PBS, (iii) 1 mg / kg+12xABC3315, (iv) 3 mg / kg PV+PBS (v) 3 mg / kg+12x ABC3315. PV was administered by injection into the retro-orbital venous sinus. ABC3315 was administered by intraperitoneal injection, divided into 5 injections, of which 30% of the dose was administered immediately after PV injection, 25% was administered 8 hours after PV injection, and 15% was administered at 24 hours, 32 hours, and 48 hours. Tumor volume and body weight were monitored daily, and the tumor volume reached 2000mm 3 Mice were sacrificed at 4 hr. Kaplan-Meier survival curves were generated in GraphPad Prism 7 and compared using the log-rank test at a significance level of p ≤ 0.05.

[0232] Toxicity studies

[0233] Toxicity evaluation of PV following simultaneous and nonsimultaneous administration of 3x ABC3315 at a dose of 120 mg / kg was contracted by Champions Oncology (Rockville, MD). Swiss Webster mice (5 per group) were injected with PV via tail vein injection. ABC3315 was administered via intraperitoneal injection with the same administration regimen as outlined in the xenograft study methods. Control mice received an equal amount of PBS each time. Mouse body weights were measured daily for 14 days following PV injection. Mice with greater than 10% weight loss were given ad libitum access to dietary gel.

[0234] Example 2

[0235] This example provides a description of the results obtained using the materials and methods of Example 1.

[0236] A scFv phage library was developed from spleen cells obtained from immunized mice and screened to identify scFvs with selective binding to free MMAE relative to vc-MMAE conjugate. Figure 1 The top 5 hits showing binding to free MMAE and negligible binding to trastuzumab-vc-MMAE (T-vc-MMAE) are shown. The bar graph represents the proportion of phage displayed scFvs bound to MMAF-peg11-biotin-streptavidin spiked with MMAE or T-vc-MMAE compared to control wells. Figure 2 The amino acid sequences of the clones are provided.

[0237] Random mutagenesis phage libraries were developed from clones 1H2, 1B3, and 3B8 and subsequently panned and screened to identify clones with increased affinity for MMAE. Figure 3 The fractional binding signals observed for individual clones from four 96-well plates with and without incubation with 10 nM or 1 nM free MMAE are shown. The clones were sequenced and the amino acid sequences were aligned ( Figure 4 ).

[0238] A humanized anti-MMAE fab (ABC3315) was generated by the remodeling protocol, where the z-score of the variable domain of the heavy chain increased from -0.624 to 0.872 and the z-score of the light chain increased from 0.120 to 0.989 after humanization. ABC3315 was characterized using a competitive ELISA in which increasing concentrations of free MMAE / MMAF and T-vc-MMAE were present ( Figure 5 ). Addition of free MMAE and MMAF reduced the binding of ABC3315 with an IC50 of ~1 nM, whereas no reduction in binding signal was observed after addition of T-vc-MMAE. For high affinity interactions, IC50 values ​​observed in competition ELISA experiments are generally greater than the equilibrium dissociation constant. To better characterize the binding affinity of ABC3315, a series of kinetic titration surface plasmon resonance (SPR) analyses were performed. The rate constant for binding of ABC3315 to MMAF-peg11-biotin bound to a neutravidin SPR chip was estimated. Figure 6 The left panel provides the observed sensorgram. The estimated equilibrium dissociation constant for ABC3315 binding to MMAF-peg11-biotin-neutravidin is 44.2 pM. Subsequent SPR experiments were performed using biotinylated ABC3315 immobilized on a streptavidin chip. Figure 6 The middle panel shows the sensorgram observed upon injection of free MMAE on immobilized ABC3315. The fitted equilibrium dissociation constant for free MMAE binding to ABC3315 is 7.7 pM. ABC3315 has a faster binding rate constant to free MMAE (2.67x105 M-1sec-1) compared to immobilized MMAF-peg11-biotin (4.11x104 M-1sec-1), thus having a higher affinity for free MMAE. Figure 6 The right panel shows the sensorgram observed after kinetic titration of polatuzumab vedotin (PV) on immobilized ABC3315. Consistent with the selective binding of ABC3315 to free MMAE, no binding signal was observed and the change in response units was minimal (±2 μRU) during the binding assay.

[0239] The Burkitt lymphoma cell line (Ramos) was incubated with free MMAE or PV, with or without 500 nM ABC3315. The cell survival fraction was determined after 4 days of incubation and calculated as the quotient of the MTT signal of the treated wells divided by the control wells. The addition of ABC3315 increased the IC50 of free MMAE in RAMOS cells from 0.12 nM to 95.96 nM. The IC50 of PV-treated cells was 0.12 nM, and the IC50 of PV+ABC3315-treated cells was 0.13 nM. Similar results were obtained in the HER2+SKBR3 cell line after treatment with free MMAE or trastuzumab-vc-MMAE. After the use of ABC3315, the IC50 of free MMAE increased from 0.09 nM to 46.24 nM, and the IC50 of T-vc-MMAE was 0.04 nM and 0.03 nM when co-cultured with ABC3315 and without co-culture, respectively. ABC3315 increased the ADC potency / payload potency ratio of PV by 738-fold in Ramos cells and by 385-fold in SKBR3 cells. Figure 7 Cell viability curves of free MMAE, PV, and T-vc-MMAE are provided.

[0240] To evaluate the effect of ABC3315 on PV efficacy in vivo, NU / J mice bearing RAMOS xenografts were injected with a single dose of PV alone or co-administered with ABC3315 at a 12-fold molar ratio relative to bound MMAE. Figure 8 A shows the tumor growth curves of each group. Figure 8 Survival curves for each group are provided in B. Using the Log-Rank test, co-treatment with ABC3315 did not significantly alter the survival of mice treated with PV at a dose of 1 mg / kg (p=0.075) or 3 mg / kg (p=0.89).

[0241] To evaluate the effect of ABC3315 on PV toxicity, Swiss-Webster mice were intravenously injected with 120 mg / kg PV together with PBS vehicle or co-administered with ABC3315 at a 3-fold molar ratio relative to bound MMAE. Mouse body weights were measured daily for 14 days after administration. Fig. 9 A provides the observed mean weight change over time for each group, Fig. 9 B provides the minimum body weight of each group. Administration of ABC3315 together with 120 mg / kg PV reduced the minimum body weight loss of mice from 11.9% (SD ± 7.0%) for mice treated with ADC plus PBS to 4.1 ± 2.1% (p = 0.045 for mice treated with ADC plus ABC3315).

[0242] Example 3

[0243] method

[0244] Albumin-bound anti-MMAE Fab

[0245] The gene sequence of a previously reported albumin binding peptide (abandoned Genentech patent application: US20050287153A1) was added to the C-terminus of the ABC3315 heavy chain. The variants were expressed in ExpiCho-S cells as recommended by the manufacturer. Human serum albumin binding affinity was assessed using the surface plasmon resonance (SPR) method. For SPR evaluation, human serum albumin (HSA) was immobilized on a carboxymethyl dextran chip. ABC3315-WEDD with a concentration range of 61.2nM to 11.4μM was injected onto the HSA chip for 2 minutes with a dissociation time of 1 minute. ABC3315-WE with a concentration range of 637nM to 30μM was injected onto the HSA chip with an injection time of 2 minutes and a dissociation time of 1 minute. The observed sensorgrams were fitted to obtain the association rate constant, dissociation rate constant, and equilibrium dissociation rate constant for HSA binding.

[0246] Albumin binding variant sequences

[0247] *Albumin binding peptide is underlined. The light chain sequence of ABC3315 was not modified for the albumin binding variant.

[0248] ABC3315-WEDD

[0249] EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCGGGSDICLPRWGCLWEDD(SEQ ID NO:51)

[0250] ABC3315-WE

[0251] EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCGGGSDICLPRWGCLWE(SEQ ID NO:52)

[0252] ABC3315 mutant

[0253] The ABC3315 variants with improved affinity or humanization have been generated. In order to identify the ABC3315 mutants with increased affinity relative to ABC3315, error-prone PCR was used to generate mutant scFv phage libraries based on ABC3315 scFv sequences. The phages displaying the scFv mutants were combined with MMAF-peg11-biotin-streptavidin magnetic beads for one hour, and the phages displaying the scFv were subsequently dissociated from the magnetic beads for 24 hours, 72 hours and 120 hours, and the 1st, 2 and 3 rounds of panning were performed, wherein the dissociation buffer contained 1 μM free ABC3315 to prevent the scFv phages from re-combining during the dissociation process. After each round of dissociation, the remaining phages displaying the scFv were eluted using 10mg / mL trypsin for 20 minutes. The phages eluted after each round were amplified, and the amplified phages were used as the input titer for the next round of panning. After the third round of panning, a small amount of phage eluate was titrated and the remaining phage was amplified. From the titrated phage, a 2-96 well master plate was generated and phage was screened using a dissociation ELISA. Briefly, 10 ug / ml streptavidin was immobilized on Immobilizer TMAmino (Thermo Scientific) plates overnight. The next day, unreacted sites were blocked with 10mM ethanolamine in pH 9.5 sodium bicarbonate buffer, followed by binding of MMAF-peg11-biotin to immobilized streptavidin. Amplified phages from each well of the 2 master plates were diluted 4-fold into PBS and added to the plates in duplicate and incubated in a shaking incubator for one hour. The plates were washed 3 times, and then PBS or PBS containing 1μM MMAF was added to the wells. The plates were incubated for 24 hours, washed 3 times, and then anti-CM13 phage secondary antibodies were added to the wells. Phages bound to 1μM MMAF were normalized to bound phages for each clone treated with PBS alone. Wells with minimal changes observed under MMAF incubation were predicted to have slower dissociation rates and were sequenced. Phagemid DNA was isolated from phages amplified after the third round of panning, digested using NdeI and XhoI restriction enzymes, scFv DNA was isolated using agarose gel electrophoresis, scFv DNA was ligated into pet22b and transformed into shuffle E. coli cells. The next day, a single transformed cell was inoculated into the inner well of an ELISA plate and cultured overnight. The next day, 10 μL of the culture medium from each well was transferred to a 96-well deep-well plate and placed in a shaker incubator at 30°C for 4 hours. 1 mM IPTG was added to each well, and the cells were incubated in a shaker incubator at 16°C overnight. The next day, the cells were lysed and diluted in PBS containing HRP-labeled ABC3315. The solution was transferred to a plate coated with MMAF-peg11-biotin-neutravidin pre-blocked with 2% milk. The plate was incubated for 2 hours, washed and the bound ABC3315-HRP was assessed after the addition of turbo TMB substrate. Clones with reduced ABC3315-HRP signal were sequenced to identify scFv clones that effectively compete with ABC3315 for substrate binding. To generate mutants with higher humanization, The murine residues in the variable heavy chain of ABC3315 that were retained during humanization (due to their location in positions that may affect MMAE binding) were mutated to amino acid residues commonly found in the human sequence using a site-directed mutagenesis kit (New England BioLabs). Mutants were expressed in ExpiCho-S cells following the manufacturer's recommendations. The binding activity of mutant ABC3315 was compared using an indirect ELISA method with MMAF-peg11-biotin-neutravidin as the capture substrate and anti-human Fab alkaline phosphatase as the detection antibody.

[0254] result

[0255] Human serum albumin binding

[0256] Both ABC3315-WEDD and ABC3315-WE were observed to bind to immobilized human serum albumin. Fig.10 and Fig.11 The observed sensorgrams for ABC3315-WEDD and ABC3315-WE are provided, respectively. The best fit association rate constant, dissociation rate constant, and equilibrium dissociation rate constant are provided in the inset of each figure. The estimated equilibrium dissociation rate constant for ABC3315-WEDD towards human serum albumin is 2.34 μM. The estimated equilibrium dissociation rate constant for ABC3315-WE towards human serum albumin is 10.50 μM. Fig.10 and Fig.11 As shown in the bottom panel of , similar values ​​of the equilibrium dissociation rate constant were obtained using the maximum signal observed at each concentration.

[0257] ABC3315 mutant

[0258] After panning and screening of the mutant scFv phage display library, unique ABC3315 mutants (e.g. Fig.12 As shown). Several clones with CDR mutations were identified, which are of great significance for the development of ABC3315 derivatives with increased MMAE binding affinity. Four clones were identified, in which the phenylalanine (F) at the 27th position of the heavy chain was mutated to leucine (L), and therefore, this substitution was selected for further characterization. The mutant ABC3315Fab with increased humanization (VH_E42G, VH_R44G, VH_L61A, VH_F37V) or potential increased MMAE affinity (VH_F27L) was compared using an indirect ELISA method. The binding signals of most ABC3315 mutants to fixed MMAF-peg11-biotin were similar to those of ABC3315, indicating that the binding function was retained after mutagenesis. The VH_F37V clone showed a greatly reduced binding signal relative to ABC3315, indicating that the phenylalanine residue at the 37th position of the heavy chain is essential for high-affinity binding. ABC3315 and VH_F27L were compared in a competitive SKBR3 cell cytotoxicity assay as described, however, the concentration of Fab was maintained at a 5-fold excess relative to MMAE for all MMAE dilutions, rather than a constant 500 nM Fab concentration. It was observed that SKBR3 cells treated with MMAE and VH_F27L had greater cell viability ( Fig.14 The observed results are consistent with VH-F27L having a higher affinity for free MMAE than ABC3315.

[0259] Mutated heavy chain sequence

[0260] VH_E42G (Humanized)

[0261]

[0262] VH_R44G (User-friendly)

[0263]

[0264] VH_L61A (User-friendly)

[0265]

[0266] VH_F37V (User-friendly)

[0267]

[0268] VH_F27L (Affinity)

[0269] EVQLVESGGGLVKPGGSLKLSCAASG L TFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC (SEQ ID NO:4)

[0270] Mutated scFv sequence

[0271] >C1B11

[0272] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYRASTRHTGVPDRFSGNGSGTDLTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFSFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS (SEQ ID NO:57)

[0273] >C1C8

[0274] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRPEWVATISSGGSYTYYLDSVKGRFTISRDNNKNTLFLQMSSLRSEDTAVYYCLASMITTDYFEYWGQGTLVTVSS(SEQ ID NO:58)

[0275] >C1D7

[0276] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTELEIKRGGSGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLVTTDYFEYWGQGTLVTVSS(SEQ ID NO:59)

[0277] >C1D10

[0278] MDIVMTQSTSSMRASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:60)

[0279] >C1E2

[0280] MDIVMTQSPSSLSASVGDRVIFTCRTSQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTELEVKRSGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSSGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLMTTDYFEYWGQGTLVTVSS(SEQ ID NO:61)

[0281] >C1G3

[0282] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQFSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKKTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:62)

[0283] >C2C2

[0284] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSGVQLVESGGGLVKPGGSLKLSCAASGFTISGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTTSRDNSINTLYLQMSSLRSEDTAVYYCLASLFTTDYFEYWGQGTLVTVNS(SEQ ID NO:63)

[0285] >C2C4

[0286] MDIVMTQSPSSLSASVGDRVTISCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGSKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:64)

[0287] >C2D11

[0288] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVQDRFSGSGSGTVFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPGKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:65)

[0289] >C2F5

[0290] MDIVMTQSPSSLGASVGDRVTITCRASQDIGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGAGTKLEIRRGGGGSGGGGSGGGGSGGGGSEVQLDESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:66)

[0291] >DBG6

[0292] MDIVMTQSPSSLSTSVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPGRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEINRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:67)

[0293] >DAD3

[0294] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVERGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLLTTDYFEYWGQGTLVTVSS(SEQ ID NO:68)

[0295] >DAD4

[0296] MDIVMTQSPSSLNASVGGRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVGTISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:69)

[0297] >DAG4

[0298] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYLEYWGQGTLVTVSS(SEQ ID NO:70)

[0299] >DAC2

[0300] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTHYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:71)

[0301] >DBF5

[0302] MDIVMTQSPSSQSASIGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIERGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQTSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:72)

[0303] >DAD10

[0304] MDIVMTQSPSGLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGSDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLFESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRPEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:73)

[0305] >AC10

[0306] MDIVMTQSPSSLSASVGDRVTITCSASQDVGTAGAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGDSGSGGSEVQLVESGGGLVKRGGSLKLSCAASGFTLSGYAMSWFRQAPGKRPEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:74)

[0307] >DBB10

[0308] MDIVMTQSPSSLSASVGDRVTFTCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:75)

[0309] >DBD2

[0310] MDIVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFNLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGGYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGALVTVSS(SEQ ID NO:76)

[0311] >DBG2

[0312] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSKVQLVESGGGLVKRGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQLSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:77)

[0313] >DAD5

[0314] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGGGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:78)

[0315] >DAG2

[0316] MDIVMTQSPSSLSASVGDRVTITCRTSQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGAGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:79)

[0317] >DAG7

[0318] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYRQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKGGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:80)

[0319] Example 4

[0320] This example includes the additional described antibody sequences, as well as the characterization of the antibody herein designated as ABC3315.

[0321] >DAB6

[0322] MDIVMTQSPSSLSASVGDRVTIACRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEVKRGGGGSGGGGSGSGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGSTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRYTISRDNSKNTLYLQMSSLRSEDTAVYYCLASRITTDYFEYWGQGTLVTVSS(SEQ ID NO:81)

[0323] >DAC3

[0324] MDIVMTQSPSSLSASVGDRVTITCRASQDVGAAVAWYKKKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:82)

[0325] >DAC4

[0326] MDIVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYSCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGRVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:83)

[0327] >DAC9

[0328] MDIVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:84)

[0329] >DAD9

[0330] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPGRFSGSGSGTDFTLTIRGLQSEDEADYFCQQYSNYPYTFGGGTKLDIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLFTTDYFEYWGQGTLVTVSS(SEQ ID NO:85)

[0331] >DAE2

[0332] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIHWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKMEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPGKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:86)

[0333] >DAG5

[0334] MDIVMTQSPSSLSASVGGRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRLTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:87)

[0335] >DAG10

[0336] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:88)

[0337] >DBC8

[0338] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGADFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMNSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:89)

[0339] >DBD5

[0340] MDIVMTQSPGSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRLSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGSTFSGYAMSWFRQAPEKRPEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:90)

[0341] >DBD9

[0342] MDIVMTQSPSSLRASVGGRVTITCRASQDVGTAVAWYQQKPGQSPKLLIHWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGSTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSINTLYLQMSSLRSEDTAVYYCLASQFTTDYFEYWGQGNLVTVSS(SEQ ID NO:91)

[0343] >DBD11

[0344] MDNVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGRSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSVDGADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:92)

[0345] >DBG3

[0346] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEINRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTLSGYAMSWFRQAREKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTPVTVSS(SEQ ID NO:93)

[0347] >DBG4

[0348] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLTYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYSCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSDGGGTGGGGSEVQLVESGGGLIKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:94)

[0349] >DBG8

[0350] MDIVMTQSLSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGNYTYYLDSVKGRFTISRDNSKNILYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:95)

[0351] >DBG11

[0352] MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQNPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYTMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKKTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:96)

[0353] ABC3317 sequence

[0354] Light chain

[0355] DIVMTQSPSSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:1)

[0356] Heavy chain (albumin binding sequence in italics)

[0357]

[0358]

[0359] ABC3320 sequence

[0360] Light chain (H55Y mutation in bold)

[0361]

[0362] Heavy chain (F27L mutation in bold, albumin binding sequence in italics)

[0363]

[0364] Newly identified sequences from a naive human Fab library

[0365] Human Fab (VL and VH listed)

[0366] >hFM1

[0367] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAYYVHWYQQVPGTAPRLLIFDNDNRPSGVPDRFSASKSGTSASLAIIGLQAEDEAEYYCQSVDYSLGDGVVFGGGTKLTVL(SEQ ID NO:177)

[0368] QVQLVESGPGLVKPSETLSLTCAVSGGSISSNNWWSWVRQTPGKGLEWIGEIYHSGSTNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCANLRGQWGQGTLVTVSS(SEQ ID NO:100)

[0369] >hFM2

[0370] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPRLLISGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGHVVFGGGTKLTVL(SEQ ID NO:101)

[0371] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGKRGNFDYWGQGTLVTVSS(SEQ ID NO:102)

[0372] >hFM3

[0373] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0374] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0375] >hFM4

[0376] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0377] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0378] >hFM5

[0379] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0380] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0381] >hFM6

[0382] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129)

[0383] QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130)

[0384] >hFM7

[0385] QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVL(SEQ ID NO:137)

[0386] QVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:138)

[0387] >hFM8

[0388] QAVLTQPSSVSGAPGQRVTISCTGSSTNIGADFDVHWYQQLPGTAPKLLIHGDNNRPSGVPDRFSGSKSGTSAYLAISGLQAEDEADYYCQTYDSRSSGSRVFGGGTKVTVL(SEQ ID NO:139)

[0389] QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107)

[0390] >hFM9

[0391] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0392] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0393] >hFM11

[0394] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131)

[0395] QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132)

[0396] >hFM12

[0397] QAVLTQPSSVSGAPGQRVAISCTGSSSNIAAGYDVQWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSVLYVFGTGTKVTVL(SEQ ID NO:156)

[0398] QLQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEIYHSGSTNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCAKRPRNSGYLGAFDIWGQGTMVTVSS(SEQ ID NO:157)

[0399] >hFM13

[0400] DVVMTQSPLSLAVTLGQPASISCRSSQSLLHSSGYKFLNWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQSPTFGGGTKVEIK(SEQ ID NO:158)

[0401] EVQLVESGGGLVQPGGSLRLSCTASGFTFSSFSMNWVRQAPGKGPEWVSAISGSGGGTYYADSVKGRFTISRDNAKNSLYLQMSSLRAEDTAVYYCARVRVPQAFDIWGQGTMVTVSS(SEQ ID NO:159)

[0402] >hFM14

[0403] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0404] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0405] >hFM15

[0406] QAVLTQPSSVSGAPGQRVTISCTGSSTNIGADFDVHWYQQLPGTAPKLLIHGDNNRPSGVPDRFSGSKSGTSAYLAISGLQAEDEADYYCQTYDSRSSGSRVFGGGTKVTVL(SEQ ID NO:139)

[0407] QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107)

[0408] >hFM16

[0409] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:177)

[0410] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTISVDTSKNQFSLKLNSVTAADTAVYYCARVGEGAFKDLGQGTLVTVSS(SEQ ID NO:178)

[0411] >hFM18

[0412] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0413] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0414] >hFM19

[0415] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117)

[0416] QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118)

[0417] >hFM20

[0418] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGRAPKLLIYGNNQRPSGVPDRFSGSTSGTSASLAITGPQAEDEADYYCQSYDSSLNGIWVFGGGTKLTVL(SEQ ID NO:119)

[0419] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEINPSGSTNYNPSLKSRVTMSLDTSKNQFSLKLRSVTAADTALYYCATRDYWGQGTLVTVSS(SEQ ID NO:120)

[0420] >hFM21

[0421] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0422] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0423] >hFM23

[0424] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0425] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0426] >hFM24

[0427] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131)

[0428] QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132)

[0429] >hFM28

[0430] QSVLTQPPSVSAAPGQKVTISCSGSSSDIGNNFVSWYQQLPGTAPKRLIYDNSKRPSGIPERFSGSKSGTSATLGITGLQTGDEADYYCGAWDTSLSAYVFGTGTKVTVL(SEQ ID NO:108)

[0431] QVQLQQWGPGLVKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARYSNYRHYYYGMDVWGQGTLVTVSS

[0432] >hFM29

[0433] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0434] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0435] >hFM30

[0436] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0437] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0438] >hFM31

[0439] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0440] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0441] >hFM32

[0442] QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155)

[0443] QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127)

[0444] >hFM33

[0445] QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYHVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLSAVLFGRGTKLTVL(SEQ ID NO:169)

[0446] QLQLQESGPGLVKPSGTLSLTCAVSGGSISSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLRGSGRWGQGTLVTVSS(SEQ ID NO:170)

[0447] >hFM34

[0448] QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYHVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLSAVLFGRGTKLTVL(SEQ ID NO:169)

[0449] QLQLQESGPGLVKPSGTLSLTCAVSGGSISSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLRGSGRWGQGTLVTVSS(SEQ ID NO:170)

[0450] >hFM36

[0451] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0452] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0453] >hFM39

[0454] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLYVFGTGTKVTVL(SEQ ID NO:145)

[0455] QVQLQESGPGLVKPSETLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLKSRVTISVDKAKNQFSLMLNSVTAADTAVYYCARGYGMDVWGQGTMVTVSS(SEQ ID NO:146)

[0456] >hFM40

[0457] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117)

[0458] QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118)

[0459] >hFM41

[0460] QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155)

[0461] QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127)

[0462] >hFM42

[0463] QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155)

[0464] QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127)

[0465] >hFM43

[0466] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117)

[0467] QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118)

[0468] >hFM44

[0469] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0470] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0471] >hFM45

[0472] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAYYVHWYQQVPGTAPRLLIFDNDNRPSGVPDRFSASKSGTSASLAIIGLQAEDEAEYYCQSVDYSLGDGVVFGGGTKLTVL(SEQ ID NO:99)

[0473] QVQLVESGPGLVKPSETLSLTCAVSGGSISSNNWWSWVRQTPGKGLEWIGEIYHSGSTNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCANLRGQWGQGTLVTVSS(SEQ ID NO:100)

[0474] >hFM46

[0475] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKVLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:148)

[0476] QVQLQQWGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQFPGKGLEWIGEMSHTGSTNYNPSFKSRVTISVDKSKNQFSLKLSPVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:147)

[0477] >hFM47

[0478] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:134)

[0479] QLQLQESGPGLVKPSETLSLTCTVSGGSISSGNYWSWVRQSPEKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCVGTLRTWFDYWGQGTLVTVSS(SEQ ID NO:105)

[0480] >hFM48

[0481] QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYYVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGVTGLQTGDEADYYCGTWDSSLSAYVFGTGTKVTVQQVQLQESGPGLVKSSETLSLICAVSGGSISSNNWWSWVRQPPGKGLEWIGEIHHSGTTINYNPSLKSRVTISVDKSKNQFSLQLNSVTPEDTAVYFCARSASGAFDIWGQGTMVTVSS(SEQ ID NO:142)

[0482] >hFM49

[0483] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVP(SEQ ID NO:143)

[0484] QVQLQESGPGLVELSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISIDRSQNQFSLKLTSMTAADTAVYYCARLYSGYGHGMDVWGQGTTVTVSS(SEQ ID NO:144)

[0485] >hFM51

[0486] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLYVFGTGTKVTVL(SEQ ID NO:145)

[0487] QLQLQESGPGLVKPSGTLSLNCAVSGVSISSTNWWSWVRQFPGKGLEWIGEINHSGTTNYNPSLKSRVTISVDTSKNQFSLQLNSVTPEDTAVYFCAQHLTVWGQGTLVTVSS(SEQ ID NO:179)

[0488] >hFM52

[0489] QAVLTQPSSLSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLISGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSFVFGTGTKVTVL(SEQ ID NO:164)

[0490] QLQLQESGPGLVEPSGTLSLTCAVSGVSISTRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISVDASKNQLSLKLTSVTVADTAVYYCSRKGVDAFDIWGQGTMVTVSS(SEQ ID NO:165)

[0491] >hFM53

[0492] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0493] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0494] >hFM57

[0495] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0496] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0497] >hFM58

[0498] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVP(SEQ ID NO:143)

[0499] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISIDRSQNQFSLKLTSMTAADTAVYYCARLYSGYGHGMDVWGQGTTVTVSS

[0500] >hFM59

[0501] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0502] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0503] >hFM60

[0504] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGARYDVHWYQQLPGGAPKLLIHSNSNRPSGVPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLNSYVFGTGTKVTVLQVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLRRGYWGQGTLVTVSS

[0505] >hFM62

[0506] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0507] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0508] >hFM64

[0509] QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNYYVAWYQQVPGAAPKLLIYDTNKRPSGIPDRFSGSKSGTSATLDITGLRTGDEADYYCGTWDSSLDTDVVFGGGTKLTVL(SEQ ID NO:167)

[0510] QVQLQESGPGLVKPSETLSLTCAVSGGSISSGSWWSWVRQAPGKGLEWIGEISHSGTTTYNPSLKSRVTISLDKSTSHLSLSLKSVTAADTAVYYCARELGGGAYDIWGQGTIVTVSS(SEQ ID NO:168)

[0511] >hFM65

[0512] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131)

[0513] QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132)

[0514] >hFM66

[0515] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0516] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0517] >hFM68

[0518] QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155)

[0519] QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127)

[0520] >hFM69

[0521] QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVL(SEQ ID NO:137)

[0522] QVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:140)

[0523] >hFM70

[0524] QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVL(SEQ ID NO:137)

[0525] QVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:140)

[0526] >hFM71

[0527] QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYYVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGVTGLQTGDEADYYCGTWDSSLSAYVFGTGTK VTVQ(SEQ ID NO:141)

[0528] QVQLQESGPGLVKSSETLSLICAVSGGSISSNNWWSWVRQPPGKGLEWIGEIHHSGTTINYNPSLKSRVTISVDKSKNQFSLQLNSVTPEDTAVYFCARSASGAFDIWGQGTMVTVSS(SEQ ID NO:142)

[0529] >hFM72

[0530] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0531] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0532] >hFM73

[0533] QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155)

[0534] QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127)

[0535] >hFM74

[0536] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0537] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0538] >hFM75

[0539] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVNWHQQFPGTAPKVLIFGDNARPSGVPDRFYASKSGTSASLTIIGVQSDDEADYYCSTWDDSLNAVVFGGGTTLTVL(SEQ ID NO:149)

[0540] QLQESGPGLVKPSGTLSLTCAVSGGSISSGNWWSWVRQPPGRGLEWIGEISHSGTINYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARRADGYFQPWGQGTLVTVSS

[0541] >hFM76

[0542] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0543] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0544] >hFM77

[0545] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129)

[0546] QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130)

[0547] >hFM78

[0548] QAVLTQPSSVSGAPGQRVAISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLRVFGGGTKLTVL(SEQ ID NO:162)

[0549] QLQLQESGPGLVKPSQTLSLTCTVSGASISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKVSSVTAADTAVYYCAREPRYWGQGTLVTVSS(SEQ ID NO:163)

[0550] >hFM80

[0551] QAVLTQPSSLSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLISGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSFVFGTGTKVTVL(SEQ ID NO:164)

[0552] QLQLQESGPGLVEPSGTLSLTCAVSGVSISTRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISVDASKNQLSLKLTSVTVADTAVYYCSRKGVDAFDIWGQGTMVTVSS(SEQ ID NO:165)

[0553] >hFM81

[0554] QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNYYVAWYQQVPGAAPKLLIYDTNKRPSGIPDRFSGSKSGTSATLDITGLRTGDEADYYCGTWDSSLDTDVVFGGGTKLTVL(SEQ ID NO:167)

[0555] QVQLQESGPGLVKPSETLSLTCAVSGGSISSGSWWSWVRQAPGKGLEWIGEISHSGTTTYNPSLKSRVTISLDKSTSHLSLSLKSVTAADTAVYYCARELGGGAYDIWGQGTIVTVSS(SEQ ID NO:168)

[0556] >hFM82

[0557] QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYHVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLSAVLFGRGTKLTVL(SEQ ID NO:169)

[0558] QLQLQESGPGLVKPSGTLSLTCAVSGGSISSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLRGSGRWGQGTLVTVSS(SEQ ID NO:170)

[0559] >hFM84

[0560] QAVLTQPSSVSGAPGQRLTISCTGSTSNIGAGYDVQWYQKLPGAAPKLLVYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEGHYYCQSYDSSLSGWVVFGGGTKLTVL(SEQ ID NO:171)

[0561] QLVQSGPGLVKPSGILSLTCAVSGGSITSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTMSVDKSKNQLSLKLSSLTAADTAVYYCARGGSSLPIWGQGTTVTVSS(SEQ ID NO:172)

[0562] >hFM85

[0563] QAVLTQPSSVSGAPGQTVTISCTGSSSNIGADYDVHWYQQLPGTAPKLLIYGNNNRPSGVPDRFSGSKSGASASLAITGLQADDEADYYCHSYDSTRSGLYIFGTGTRVIV(SEQ ID NO:173)

[0564] QLQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTISVDKSKNQFSLKLSSVTAEDTAVYYCTTGSSGYWGQGTLVTVSS(SEQ ID NO:174)

[0565] >hFM87

[0566] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGADYDVQWYQQLPGTAPKLLIYANNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLRVFGGGTKLTVL(SEQ ID NO:175)

[0567] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARSGRRSSWIDYWGQGTLVTVSS(SEQ ID NO:176)

[0568] >hFM88

[0569] QAVLTQPSSVSAAPGQKVTISCSGSDSNIGNYYVWWYQQLPGAAPKLLIYDNHRRPSGVPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDGSLTGYVFGPGTKVTVL(SEQ ID NO:125)

[0570] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRATISVDTSKNQFSLKLSSVTAADTAVYYCARGRVRGRWLPYYWGQGTLVTVSS(SEQ ID NO:126)

[0571] >hFM90

[0572] QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVL(SEQ ID NO:137)

[0573] QVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:140)

[0574] >hFM91

[0575] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0576] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0577] >hFM92

[0578] DIQLTQSPSSLSASVGDRVTITCRASQNINNFLNWYQQKPGNVPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSPLSTFGQGTKVEIK(SEQ ID NO:160)QVQLVQSGAEVKKPGSSVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGRINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARSVSVVTDAFDIWGQGTTVTVSS(SEQ ID NO:161)

[0579] >hFM93

[0580] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0581] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0582] >hFM94

[0583] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:153)

[0584] QVQLQESGPGLVKPSGTLSLTCDVSGGSISSNNWWSWVRQSPGKGLEWIGEIIHTGRTNYNPSLTSRVTILIDKSKNQFSLKLTSVTPEDTALYYCARLRGPFDIWGQGTMVTVSS(SEQ ID NO:154)

[0585] >hFM96

[0586] QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155)

[0587] QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127)

[0588] >hFM97

[0589] QAVLTQPSSVSGAPGQRVTISCTGSSTNIGADFDVHWYQQLPGTAPKLLIHGDNNRPSGVPDRFSGSKSGTSAYLAISGLQAEDEADYYCQTYDSRSSGSRVFGGGTKVTVL(SEQ ID NO:139)

[0590] QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107)

[0591] >hFM98

[0592] DVVMTQSPLSLAVTLGQPASISCRSSQSLLHSSGYKFLNWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQSPTFGGGTKVEIK(SEQ ID NO:158)

[0593] EVQLVESGGGLVQPGGSLRLSCTASGFTFSSFSMNWVRQAPGKGPEWVSAISGSGGGTYYADSVKGRFTISRDNAKNSLYLQMSSLRAEDTAVYYCARVRVPQAFDIWGQGTMVTVSS(SEQ ID NO:159)

[0594] >hFM99

[0595] QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121)

[0596] QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122)

[0597] >hFM100

[0598] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYEVHWYQQLPGRAPRLLIFDNNNRPSGVPDRFSASKSGTSASLAITGLRAEDEGDYYCQSYDSKRTPPYVFGTGTRVTVL(SEQ ID NO:123)

[0599] QVQLQQSGAEVKKPGASVKVSCKASGYSFSKYGMSWVRQAPGQGLEWMGWINAGNGDTKYSQKFQGRVTITRDTSASTAYMELSSLKYEDTAIYYCARRLSYYGMDVWGQGTTVTVSS(SEQ ID NO:124)

[0600] >hFM101

[0601] QAVLTQPSSVSAAPGQKVTISCSGSDSNIGNYYVWWYQQLPGAAPKLLIYDNHRRPSGVPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDGSLTGYVFGPGTKVTVL(SEQ ID NO:125)

[0602] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRATISVDTSKNQFSLKLSSVTAADTAVYYCARGRVRGRWLPYYWGQGTLVTVSS(SEQ ID NO:126)

[0603] >hFM102

[0604] QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155)QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNY

[0605] NPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127)

[0606] >hFM103

[0607] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0608] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0609] >hFM104

[0610] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGFWVFGGGTKLTVL(SEQ ID NO:112)

[0611] QVQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDNSKNQFSLQLSSVTAADTAVYYCVSNASGRRGFAWGQGTLVTVSS(SEQ ID NO:113)

[0612] >hFM105

[0613] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVNWHQQFPGTAPKVLIFGDNARPSGVPDRFYASKSGTSASLTIIGVQSDDEADYYCSTWDDSLNAVVFGGGTTLTVL(SEQ ID NO:149)

[0614] VQLQESGPGLVKPSGTLSLTCAVSGGSISSGNWWSWVRQPPGRGLEWIGEISHSGTINYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARRADGYFQPWGQGTLVTVSS(SEQ ID NO:150)

[0615] >hFM106

[0616] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKVLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:148)

[0617] QVQLQQWGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQFPGKGLEWIGEMSHTGSTNYNPSFKSRVTISVDKSKNQFSLKLSPVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:147)

[0618] >hFM107

[0619] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0620] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0621] >hFM108

[0622] VLTQPPSVSGAPGQRVTISCTGSDSNIGAGYDVHWYQQYPGIAPKLLIYAHHKRPSGVPDRFSGSTSGTSASLAITGLQAEDEADYYCQSYDSSLSGHYVFGTGTQVSVL(SEQ ID NO:151)

[0623] QVQLQESGPGLVKPSETLSLTCAVSGGSISSNNWWSWVRQTPGKGLEWIGEIYHSGNTNYNPSLKSRVTISVDKSKNQFSLKLNSVTAADTAVYYCARGRQGAFDPWGQGTLVTVSS(SEQ ID NO:152)

[0624] >hFM109

[0625] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129)

[0626] QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130)

[0627] >hFM110

[0628] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGFWVFGGGTKLTVL(SEQ ID NO:112)

[0629] QVQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDNSKNQFSLQLSSVTAADTAVYYCVSNASGRRGFAWGQGTLVTVSS(SEQ ID NO:113)

[0630] >hFM112

[0631] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0632] QVQLQESGPGLVELSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:114)

[0633] >hFM113

[0634] QSVLTQPPSVSGAPGQRITISCTGSSSNIGAGYDVQWYQQVPGKAPKHLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYFCQSYDSSLSGYVVFGGGTKLTVL(SEQ ID NO:115)

[0635] QVQLQESGPGLVKPSETLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHTGSPNYNPSLASRVTISMDKSKNQFSLNLRSVTAADTSVYYCARYGRGAFDIWGQGTMVTVSS(SEQ ID NO:116)

[0636] >hFM114

[0637] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117)

[0638] QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118)

[0639] >hFM115

[0640] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0641] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0642] >hFM116

[0643] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:153)

[0644] QVQLQESGPGLVKPSGTLSLTCDVSGGSISSNNWWSWVRQSPGKGLEWIGEIIHTGRTNYNPSLTSRVTILIDKSKNQFSLKLTSVTPEDTALYYCARLRGPFDIWGQGTMVTVSS(SEQ ID NO:154)

[0645] >hFM117

[0646] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117)

[0647] QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118)

[0648] >hFM118

[0649] VLTQPPSVSGAPGQRVTISCTGGSTNIGAGYDVHWYQQLPGTAPKLLIYGNNNRPSGVPDRFSGSQSGASASLAITGLQADDEADYYCQSYDSRLDGSKVFGTGTKVTVL(SEQ ID NO:135)

[0650] QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEMSHSGIPNYNPSLESRVTISLDKSKNQFSLILRSVTAADTAMYYCVGGSGSYSYWGQGTLVTVSS(SEQ ID NO:136)

[0651] >hFM120

[0652] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGFWVFGGGTKLTVL(SEQ ID NO:112)

[0653] QVQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDNSKNQFSLQLSSVTAADTAVYYCVSNASGRRGFAWGQGTLVTVSS(SEQ ID NO:113)

[0654] >hFM121

[0655] QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128)

[0656] QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111)

[0657] >hFM122

[0658] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0659] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0660] >hFM123

[0661] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129)

[0662] QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130)

[0663] >hFM124

[0664] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131)

[0665] QVQLQQSGPGLVKPFGRPCPLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:133)

[0666] >hFM125

[0667] QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131)

[0668] QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132)

[0669] >hFM126

[0670] QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103)

[0671] QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104)

[0672] >hFM127

[0673] QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:134)

[0674] QLQLQESGPGLVKPSETLSLTCTVSGGSISSGNYWSWVRQSPEKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCVGTLRTWFDYWGQGTLVTVSS(SEQ ID NO:105)

[0675] >hFM128

[0676] QAVLTQPSSVSGAPGQRVTISCTGSSTNIGAGFDVHWYQQLPGTAPKLLIYGDKNRPSGVPDRFSGSKSGTSAYLAITGLQAEDEADYYCQTYDSRLSGSKVFGGGTKVTVL(SEQ ID NO:106)

[0677] QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISSVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107)

[0678] >hFM130

[0679] QSVLTQPPSVSAAPGQKVTISCSGSSSDIGNNFVSWYQQLPGTAPKRLIYDNSKRPSGIPERFSGSKSGTSATLGITGLQTGDEADYYCGAWDTSLSAYVFGTGTKVTVL(SEQ ID NO:108)

[0680] QVQLQQWGPGLVKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISSVDTSKNQFSLKLSSVTAADTAVYYCARYSNYRHYYYGMDVWGQGTLVTVSS

[0681] Although the invention has been described by way of illustrative embodiments, routine modifications will be apparent to those skilled in the art and are intended to fall within the scope of this disclosure.

Claims

1. A binding partner that specifically binds to a drug used as the drug component of an antibody-drug conjugate (ADC).

2. The binding partner of claim 1, wherein the binding partner preferentially binds the drug removed from the ADC relative to the binding of the binding partner to the drug when the drug is present in the ADC.

3. The binding partner of claim 2, wherein the binding partner comprises an antibody or an antigen-binding fragment thereof, and wherein the antigen-binding fragment is optionally Fab, Fab' or F(ab')2.

4. The binding partner of claim 3, wherein the drug is monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF).

5. The binding partner of claim 4, wherein the binding partner comprises a light chain and a heavy chain, the light chain comprising the sequence: DIVMTQSPSSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:1) and the heavy chain comprises the sequence: EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGT LVTVSSASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:2) in, Optionally the sequence of said light chain and / or said heavy chain comprises at least one amino acid substitution, wherein the at least one amino acid substitution in the light chain is selected from H55Y, R24H and L60A; And / or wherein the at least one amino acid substitution in the sequence of the heavy chain is selected from F27L, R44G, E42D, E42G, L61A and I101F.

6. The binding partner of claim 5, wherein the light chain comprises a H55Y amino acid substitution or wherein the heavy chain comprises a F27L mutation.

7. The binding partner of claim 6, wherein the light chain comprises a H55Y amino acid substitution, and wherein the heavy chain comprises a F27L mutation.

8. The binding ligand of claim 7, wherein the light chain comprises the sequence DIVMTQSPSSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:3) And the heavy chain comprises the sequence: EVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGT LVTVSSASTKGPSVFPLAPSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:4).

9. The binding partner of claim 8, wherein the heavy chain or the light chain or a combination thereof is adapted to further bind to a substance other than the drug in the blood of a subject to which the binding partner is administered.

10. The binding partner of claim 9, wherein binding of the binding partner to the substance in the blood increases the half-life of the binding partner in the circulation of the individual, and wherein optionally the heavy chain is adapted to bind to the substance.

11. A method for reducing off-target toxicity of an antibody-drug conjugate (ADC), comprising administering the binding partner of any one of claims 1 to 10 to an individual who has received or is concurrently receiving the ADC.

12. The method of claim 11, wherein the binding partner preferentially binds the drug removed from the ADC relative to the binding of the binding partner to the drug when the drug is present in the ADC.

13. The method of claim 12, wherein the binding partner comprises an antibody or an antigen-binding fragment thereof, and wherein the antigen-binding fragment is optionally Fab, Fab' or F(ab')2.

14. The method of claim 13, wherein the drug is monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF).

15. The method of claim 14, wherein the binding partner comprises a light chain and a heavy chain, the light chain comprising the sequence: DIVMTQSPSSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:97) and the heavy chain comprises the sequence: EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGT LVTVSSASTKGPSVFPLAPSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:2) in, Optionally the sequence of said light chain and / or said heavy chain comprises at least one amino acid substitution, wherein the at least one amino acid substitution in the light chain is selected from H55Y, R24H and L60A; and / or the at least one amino acid substitution in the sequence of the heavy chain is selected from F27L, R44G, E42D, E42G, L61A and I101F.

16. The method of claim 15, wherein the light chain comprises a H55Y amino acid substitution or wherein the heavy chain comprises a F27L mutation.

17. The method of claim 16, wherein the light chain comprises a H55Y amino acid substitution, and wherein the heavy chain comprises a F27L mutation.

18. The method of claim 17, wherein the light chain comprises the sequence DIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 3) And the heavy chain comprises the sequence: EVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGT LVTVSSASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:4).

19. The method of claim 1, wherein the heavy chain or the light chain, or a combination thereof, is adapted to further bind to a substance other than the drug in a subject to whom the binding partner is administered.

20. The method of claim 19, wherein binding of the binding partner to the substance increases the half-life of the binding partner in the circulation of the individual, and wherein optionally the heavy chain is adapted to bind to the substance.

21. A binding partner comprising or consisting of a pair of light chains and heavy chains, wherein the sequences of the light chain and the heavy chain are selected from the following sequences: VH_E42G (human) EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAP G KRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:53) VH_R44G(human) EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEK G LEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:54) >VH_L60A(human) EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYADSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGT LVTVSSASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:55) >VH_F37V(human) EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWVRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGT LVTVSSASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:56) VH_F27L(affinity) EVQLVESGGGLVKPGGSLKLSCAASG L TFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:4) >C1B11 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYRASTRHTGVPDRFSGNGSGTDLTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFSFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:57) >C1C8 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRPEWVATISSGGSYTYYLDSVKGRFTISRDNNKNTLFLQMSSLRSEDTAVYYCLASMITTDYFEYWGQGTLVTVSS(SEQ ID NO:58) >C1D7 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTELEIKRGGSGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLVTTDYFEYWGQGTLVTVSS(SEQ ID NO:59) >C1D10 MDIVMTQSTSSMRASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:60) >C1E2 MDIVMTQSPSSLSASVGDRVIFTCRTSQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTELEVKRSGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSSGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLMTTDYFEYWGQGTLVTVSS(SEQ ID NO:61) >C1G3 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQFSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKKTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:62) >C2C2 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSGVQLVESGGGLVKPGGSLKLSCAASGFTISGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTTSRDNSINTLYLQMSSLRSEDTAVYYCLASLFTTDYFEYWGQGTLVTVNS(SEQ ID NO:63) >C2C4 MDIVMTQSPSSLSASVGDRVTISCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGSKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQTPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:64) >C2D11 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVQDRFSGSGSGTVFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPGKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:65) >C2F5 MDIVMTQSPSSLGASVGDRVTITCRASQDIGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGAGTKLEIRRGGGGSGGGGSGGGGSGGGGSEVQLDESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:66) >DBG6 MDIVMTQSPSSLSTSVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPGRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEINRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTVSRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:67) >DAD3 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVERGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLLTTDYFEYWGQGTLVTVSS(SEQ ID NO:68) >DAD4 MDIVMTQSPSSLNASVGGRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVGTISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:69) >DAG4 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYLEYWGQGTLVTVSS(SEQ ID NO:70) >DAC2 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTHYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:71) >DBF5 MDIVMTQSPSSQSASIGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIERGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQTSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:72) >DAD10 MDIVMTQSPSGLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGSDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLFESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRPEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:73) >AC10 MDIVMTQSPSSLSASVGDRVTITCSASQDVGTAGAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGDSGSGGSEVQLVESGGGLVKRGGSLKLSCAASGFTLSGYAMSWFRQAPGKRPEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:74) >DBB10 MDIVMTQSPSSLSASVGDRVTFTCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:75) >DBD2 MDIVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFNLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGGYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGALVTVSS(SEQ ID NO:76) >DBG2 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSKVQLVESGGGLVKRGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQLSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:77) >DAD5 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGGGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:78) >DAG2 MDIVMTQSPSSLSASVGDRVTITCRTSQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGAGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:79) >DAG7 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYRQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKGGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:80) >DAB6 MDIVMTQSPSSLSASVGDRVTIACRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEVKRGGGGSGGGGSGSGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGSTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRYTISRDNSKNTLYLQMSSLRSEDTAVYYCLASRITTDYFEYWGQGTLVTVSS(SEQ ID NO:81) >DAC3 MDIVMTQSPSSLSASVGDRVTITCRASQDVGAAVAWYKKKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:82) >DAC4 MDIVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYSCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGRVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:83) >DAC9 MDIVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:84) >DAD9 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPGRFSGSGSGTDFTLTIRGLQSEDEADYFCQQYSNYPYTFGGGTKLDIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPDKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLFTTDYFEYWGQGTLVTVSS(SEQ ID NO:85) >DAE2 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIHWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKMEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPGKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:86) >DAG5 MDIVMTQSPSSLSASVGGRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRLTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:87) >DAG10 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:88) >DBC8 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGADFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMNSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:89) >DBD5 MDIVMTQSPGSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRLSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGSTFSGYAMSWFRQAPEKRPEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:90) >DBD9 MDIVMTQSPSSLRASVGGRVTITCRASQDVGTAVAWYQQKPGQSPKLLIHWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGSTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSINTLYLQMSSLRSEDTAVYYCLASQFTTDYFEYWGQGNLVTVSS(SEQ ID NO:91) >DBD11 MDNVMTQSPSSLSASVGDRVTITCHASQDVGTAVAWYQQKPGRSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSVDGADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:92) >DBG3 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEINRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTLSGYAMSWFRQAREKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTPVTVSS(SEQ ID NO:93) >DBG4 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLTYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYSCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSDGGGTGGGGSEVQLVESGGGLIKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:94) >DBG8 MDIVMTQSLSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGNYTYYLDSVKGRFTISRDNSKNILYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:95) >DBG11 MDIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQNPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVKPGGSLKLSCAASGFTFSGYTMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKKTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSS(SEQ ID NO:96) ABC3317 sequence Light chain DIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:97) Heavy chain EVQLVESGGGLVKPGGSLKLSCAASGFTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCGGGSRLIEDICLPRWGCLWEDD(SEQ ID NO:98) >ABC3320 Light chain DIVMTQSPSSLSASVGDRVTITCRASQDVGTAVAWYQQKPGQSPKLLIYWASTRYTGVPDRFSGSGSGTDFTLTISGLQSEDEADYFCQQYSNYPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:3) Heavy chain EVQLVESGGGLVKPGGSLKLSCAASGLTFSGYAMSWFRQAPEKRLEWVATISSGGSYTYYLDSVKGRFTISRDNSKNTLYLQMSSLRSEDTAVYYCLASLITTDYFEYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC(SEQ ID NO:4) >hFM1 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAYYVHWYQQVPGTAPRLLIFDNDNRPSGVPDRFSASKSGTSASLAIIGLQAEDEAEYYCQSVDYSLGDGVVFGGGTKLTVL(SEQ ID NO:99) QVQLVESGPGLVKPSETLSLTCAVSGGSISSNNWWSWVRQTPGKGLEWIGEIYHSGSTNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCANLRGQWGQGTLVTVSS(SEQ ID NO:100) >hFM2 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPRLLISGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGHVVFGGGTKLTVL(SEQ ID NO:101) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGKRGNFDYWGQGTLVTVSS(SEQ ID NO:102) >hFM3 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM4 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM5 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM6 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129) QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130) >hFM7 QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVL(SEQ ID NO:137) QVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:140) >hFM8 QAVLTQPSSVSGAPGQRVTISCTGSSTNIGADFDVHWYQQLPGTAPKLLIHGDNNRPSGVPDRFSGSKSGTSAYLAISGLQAEDEADYYCQTYDSRSSGSRVFGGGTKVTVL(SEQ ID NO:139) QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107) >hFM9 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM11 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131) QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132) >hFM12 QAVLTQPSSVSGAPGQRVAISCTGSSSNIAAGYDVQWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSVLYVFGTGTKVTVL(SEQ ID NO:156) QLQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEIYHSGSTNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCAKRPRNSGYLGAFDIWGQGTMVTVSS(SEQ ID NO:157) >hFM13 DVVMTQSPLSLAVTLGQPASISCRSSQSLLHSSGYKFLNWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQSPTFGGGTKVEIK(SEQ ID NO:158) EVQLVESGGGLVQPGGSLRLSCTASGFTFSSFSMNWVRQAPGKGPEWVSAISGSGGGTYYADSVKGRFTISRDNAKNSLYLQMSSLRAEDTAVYYCARVRVPQAFDIWGQGTMVTVSS(SEQ ID NO:159) >hFM14 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM15 QAVLTQPSSVSGAPGQRVTISCTGSSTNIGADFDVHWYQQLPGTAPKLLIHGDNNRPSGVPDRFSGSKSGTSAYLAISGLQAEDEADYYCQTYDSRSSGSRVFGGGTKVTVL(SEQ ID NO:139) QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107) >hFM16 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:177) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTISVDTSKNQFSLKLNSVTAADTAVYYCARVGEGAFKDLGQGTLVTVSS(SEQ ID NO:178) >hFM18 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM19 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117) QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118) >hFM20 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGRAPKLLIYGNNQRPSGVPDRFSGSTSGTSASLAITGPQAEDEADYYCQSYDSSLNGIWVFGGGTKLTVL(SEQ ID NO:119) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEINPSGSTNYNPSLKSRVTMSLDTSKNQFSLKLRSVTAADTALYYCATRDYWGQGTLVTVSS(SEQ ID NO:120) >hFM21 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM23 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM24 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131) QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132) >hFM28 QSVLTQPPSVSAAPGQKVTISCSGSSSDIGNNFVSWYQQLPGTAPKRLIYDNSKRPSGIPERFSGSKSGTSATLGITGLQTGDEADYYCGAWDTSLSAYVFGTGTKVTVL(SEQ ID NO:108) QVQLQQWGPGLVKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARYSNYRHYYYGMDVWGQGTLVTVSS(SEQ ID NO:109) >hFM29 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:110) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM30 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM31 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM32 QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127) >hFM33 QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYHVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLSAVLFGRGTKLTVL(SEQ ID NO:169) QLQLQESGPGLVKPSGTLSLTCAVSGGSISSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLRGSGRWGQGTLVTVSS(SEQ ID NO:170) >hFM34 QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYHVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLSAVLFGRGTKLTVL(SEQ ID NO:169) QLQLQESGPGLVKPSGTLSLTCAVSGGSISSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLRGSGRWGQGTLVTVSS(SEQ ID NO:170) >hFM36 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM39 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLYVFGTGTKVTVL(SEQ ID NO:145) QVQLQESGPGLVKPSETLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLKSRVTISVDKAKNQFSLMLNSVTAADTAVYYCARGYGMDVWGQGTMVTVSS(SEQ ID NO:146) >hFM40 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117) QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118) >hFM41 QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127) >hFM42 QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127) >hFM43 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117) QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118) >hFM44 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM45 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAYYVHWYQQVPGTAPRLLIFDNDNRPSGVPDRFSASKSGTSASLAIIGLQAEDEAEYYCQSVDYSLGDGVVFGGGTKLTVL(SEQ ID NO:99) QVQLVESGPGLVKPSETLSLTCAVSGGSISSNNWWSWVRQTPGKGLEWIGEIYHSGSTNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCANLRGQWGQGTLVTVSS(SEQ ID NO:100) >hFM46 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKVLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:148) QVQLQQWGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQFPGKGLEWIGEMSHTGSTNYNPSFKSRVTISVDKSKNQFSLKLSPVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:147) >hFM47 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:134) QLQLQESGPGLVKPSETLSLTCTVSGGSISSGNYWSWVRQSPEKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCVGTLRTWFDYWGQGTLVTVSS(SEQ ID NO:105) >hFM48 QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYYVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGVTGLQTGDEADYYCGTWDSSLSAYVFGTGTKVTVQ QVQLQESGPGLVKSSETLSLICAVSGGSISSNNWWSWVRQPPGKGLEWIGEIHHSGTTINYNPSLKSRVTISVDKSKNQFSLQLNSVTPEDTAVYFCARSASGAFDIWGQGTMVTVSS(SEQ ID NO:142) >hFM49 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVP(SEQ ID NO:143) QVQLQESGPGLVELSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISIDRSQNQFSLKLTSMTAADTAVYYCARLYSGYGHGMDVWGQGTTVTVSS(SEQ ID NO:144) >hFM51 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLYVFGTGTKVTVL(SEQ ID NO:145) QLQLQESGPGLVKPSGTLSLNCAVSGVSISSTNWWSWVRQFPGKGLEWIGEINHSGTTNYNPSLKSRVTISVDTSKNQFSLQLNSVTPEDTAVYFCAQHLTVWGQGTLVTVSS(SEQ ID NO:179) >hFM52 QAVLTQPSSLSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLISGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSFVFGTGTKVTVL(SEQ ID NO:164) QLQLQESGPGLVEPSGTLSLTCAVSGVSISTRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISVDASKNQLSLKLTSVTVADTAVYYCSRKGVDAFDIWGQGTMVTVSS(SEQ ID NO:165) >hFM53 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM57 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM58 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVP(SEQ ID NO:143) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISIDRSQNQFSLKLTSMTAADTAVYYCARLYSGYGHGMDVWGQGTTVTVSS >hFM59 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM60 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGARYDVHWYQQLPGGAPKLLIHSNSNRPSGVPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLNSYVFGTGTKVTVLQVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCASLRRGYWGQGTLVTVSS>hFM62 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM64 QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNYYVAWYQQVPGAAPKLLIYDTNKRPSGIPDRFSGSKSGTSATLDITGLRTGDEADYYCGTWDSSLDTDVVFGGGTKLTVL(SEQ ID NO:167) QVQLQESGPGLVKPSETLSLTCAVSGGSISSGSWWSWVRQAPGKGLEWIGEISHSGTTTYNPSLKSRVTISLDKSTSHLSLSLKSVTAADTAVYYCARELGGGAYDIWGQGTIVTVSS(SEQ ID NO:168) >hFM65 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131) QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132) >hFM66 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM68 QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127) >hFM69 QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVLQVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:140) >hFM70 QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVLQVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:140) >hFM71 QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYYVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGVTGLQTGDEADYYCGTWDSSLSAYVFGTGTKVTVQ QVQLQESGPGLVKSSETLSLICAVSGGSISSNNWWSWVRQPPGKGLEWIGEIHHSGTTINYNPSLKSRVTISVDKSKNQFSLQLNSVTPEDTAVYFCARSASGAFDIWGQGTMVTVSS(SEQ ID NO:142) >hFM72 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM73 QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127) >hFM74 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM75 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVNWHQQFPGTAPKVLIFGDNARPSGVPDRFYASKSGTSASLTIIGVQSDDEADYYCSTWDDSLNAVVFGGGTTLTVL(SEQ ID NO:149) QLQESGPGLVKPSGTLSLTCAVSGGSISSGNWWSWVRQPPGRGLEWIGEISHSGTINYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARRADGYFQPWGQGTLVTVSS >hFM76 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM77 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129) QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130) >hFM78 QAVLTQPSSVSGAPGQRVAISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLRVFGGGTKLTVL(SEQ ID NO:162) QLQLQESGPGLVKPSQTLSLTCTVSGASISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKVSSVTAADTAVYYCAREPRYWGQGTLVTVSS(SEQ ID NO:163) >hFM80 QAVLTQPSSLSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLISGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSFVFGTGTKVTVL(SEQ ID NO:164) QLQLQESGPGLVEPSGTLSLTCAVSGVSISTRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISVDASKNQLSLKLTSVTVADTAVYYCSRKGVDAFDIWGQGTMVTVSS(SEQ ID NO:165) QLQLQESGPGLVEPSGTLSLTCAVSGVSISTRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSFVSRVTISVDASKNQLSLKLTSVTVADTAVYYCSRKGVDAFDIWGQGTMVTVS(SEQ ID NO:166) >hFM81 QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNYYVAWYQQVPGAAPKLLIYDTNKRPSGIPDRFSGSKSGTSATLDITGLRTGDEADYYCGTWDSSLDTDVVFGGGTKLTVL(SEQ ID NO:167) QVQLQESGPGLVKPSETLSLTCAVSGGSISSGSWWSWVRQAPGKGLEWIGEISHSGTTTYNPSLKSRVTISLDKSTSHLSLSLKSVTAADTAVYYCARELGGGAYDIWGQGTIVTVSS(SEQ ID NO:168) >hFM82 QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNYHVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDSSLSAVLFGRGTKLTVL(SEQ ID NO:169) QLQLQESGPGLVKPSGTLSLTCAVSGGSISSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARLRGSGRWGQGTLVTVSS(SEQ ID NO:170) >hFM84 QAVLTQPSSVSGAPGQRLTISCTGSTSNIGAGYDVQWYQKLPGAAPKLLVYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEGHYYCQSYDSSLSGWVVFGGGTKLTVL(SEQ ID NO:171) QLVQSGPGLVKPSGILSLTCAVSGGSITSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTMSVDKSKNQLSLKLSSLTAADTAVYYCARGGSSLPIWGQGTTVTVSS(SEQ ID NO:172) >hFM85 QAVLTQPSSVSGAPGQTVTISCTGSSSNIGADYDVHWYQQLPGTAPKLLIYGNNNRPSGVPDRFSGSKSGASASLAITGLQADDEADYYCHSYDSTRSGLYIFGTGTRVIV(SEQ ID NO:173) QLQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTISVDKSKNQFSLKLSSVTAEDTAVYYCTTGSSGYWGQGTLVTVSS(SEQ ID NO:174) >hFM87 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGADYDVQWYQQLPGTAPKLLIYANNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLRVFGGGTKLTVL(SEQ ID NO:175) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARSGRRSSWIDYWGQGTLVTVSS(SEQ ID NO:176) >hFM88 QAVLTQPSSVSAAPGQKVTISCSGSDSNIGNYYVWWYQQLPGAAPKLLIYDNHRRPSGVPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDGSLTGYVFGPGTKVTVL(SEQ ID NO:125) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRATISVDTSKNQFSLKLSSVTAADTAVYYCARGRVRGRWLPYYWGQGTLVTVSS(SEQ ID NO:126) >hFM90 QAVLTQPSSVSGAPGQRVTISCTGTNSNIGAGYDVHWYQQFPGAAPKVLIYGNTNRPSGVPGRFSASKSGTSASLAITGLQAEDEADYYCQTYDSSLSGTWVFGGGTKLTVLQVQLQQWGPGLVKPSGTLSLTCAVSGVSISSSNWWSWVRQTPGKGLEWIGEISHSGSPNYNPSLKSRVTISVDTSKNQFSLKLTSVTAADSAVYYCARGGGPIPHIWGQGTTVTVSS(SEQ ID NO:140) >hFM91 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM92 DIQLTQSPSSLSASVGDRVTITCRASQNINNFLNWYQQKPGNVPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSPLSTFGQGTKVEIK(SEQ ID NO:160) QVQLVQSGAEVKKPGSSVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGRINPNSGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARSVSVVTDAFDIWGQGTTVTVSS(SEQ ID NO:161) >hFM93 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM94 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:153) QVQLQESGPGLVKPSGTLSLTCDVSGGSISSNNWWSWVRQSPGKGLEWIGEIIHTGRTNYNPSLTSRVTILIDKSKNQFSLKLTSVTPEDTALYYCARLRGPFDIWGQGTMVTVSS(SEQ ID NO:154) >hFM96 QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127) >hFM97 QAVLTQPSSVSGAPGQRVTISCTGSSTNIGADFDVHWYQQLPGTAPKLLIHGDNNRPSGVPDRFSGSKSGTSAYLAISGLQAEDEADYYCQTYDSRSSGSRVFGGGTKVTVL(SEQ ID NO:139) QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107) >hFM98 DVVMTQSPLSLAVTLGQPASISCRSSQSLLHSSGYKFLNWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQSPTFGGGTKVEIK(SEQ ID NO:158) EVQLVESGGGLVQPGGSLRLSCTASGFTFSSFSMNWVRQAPGKGPEWVSAISGSGGGTYYADSVKGRFTISRDNAKNSLYLQMSSLRAEDTAVYYCARVRVPQAFDIWGQGTMVTVSS(SEQ ID NO:159) >hFM99 QAVLTQPSSVSGAPGQRVTISCTGSRSNIGANYDVHWYQQLPGTAPKLLIFNDNIRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:121) QVQLQESGPGLVKPSGTLSLTCAVSGVSISTTNWWSWVRQPPGKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCASYWLGNFDYWGQGTLVTVSS(SEQ ID NO:122) >hFM100 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYEVHWYQQLPGRAPRLLIFDNNNRPSGVPDRFSASKSGTSASLAITGLRAEDEGDYYCQSYDSKRTPPYVFGTGTRVTVL(SEQ ID NO:123) QVQLQQSGAEVKKPGASVKVSCKASGYSFSKYGMSWVRQAPGQGLEWMGWINAGNGDTKYSQKFQGRVTITRDTSASTAYMELSSLKYEDTAIYYCARRLSYYGMDVWGQGTTVTVSS(SEQ ID NO:124) >hFM101 QAVLTQPSSVSAAPGQKVTISCSGSDSNIGNYYVWWYQQLPGAAPKLLIYDNHRRPSGVPDRFSGSKSGTSATLDITGLQTGDEADYYCGTWDGSLTGYVFGPGTKVTVL(SEQ ID NO:125) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRATISVDTSKNQFSLKLSSVTAADTAVYYCARGRVRGRWLPYYWGQGTLVTVSS(SEQ ID NO:126) >hFM102 QAVLTQPSSVSGAPGQRVIISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGNYVFGPGTKVTVL(SEQ ID NO:155) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEISHSGITNYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:127) >hFM103 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM104 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGFWVFGGGTKLTVL(SEQ ID NO:112) QVQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDNSKNQFSLQLSSVTAADTAVYYCVSNASGRRGFAWGQGTLVTVSS(SEQ ID NO:113) >hFM105 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVNWHQQFPGTAPKVLIFGDNARPSGVPDRFYASKSGTSASLTIIGVQSDDEADYYCSTWDDSLNAVVFGGGTTLTVL(SEQ ID NO:149) VQLQESGPGLVKPSGTLSLTCAVSGGSISSGNWWSWVRQPPGRGLEWIGEISHSGTINYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARRADGYFQPWGQGTLVTVSS(SEQ ID NO:150) >hFM106 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKVLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSVVFGGGTKLTVL(SEQ ID NO:148) QVQLQQWGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQFPGKGLEWIGEMSHTGSTNYNPSFKSRVTISVDKSKNQFSLKLSPVTAADTAVYYCARGSTGAFDVWGQGTMVTVSS(SEQ ID NO:147) >hFM107 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM108 VLTQPPSVSGAPGQRVTISCTGSDSNIGAGYDVHWYQQYPGIAPKLLIYAHHKRPSGVPDRFSGSTSGTSASLAITGLQAEDEADYYCQSYDSSLSGHYVFGTGTQVSVL(SEQ ID NO:151) QVQLQESGPGLVKPSETLSLTCAVSGGSISSNNWWSWVRQTPGKGLEWIGEIYHSGNTNYNPSLKSRVTISVDKSKNQFSLKLNSVTAADTAVYYCARGRQGAFDPWGQGTLVTVSS(SEQ ID NO:152) >hFM109 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129) QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130) >hFM110 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGFWVFGGGTKLTVL(SEQ ID NO:112) QVQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDNSKNQFSLQLSSVTAADTAVYYCVSNASGRRGFAWGQGTLVTVSS(SEQ ID NO:113) >hFM112 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVELSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:114) >hFM113 QSVLTQPPSVSGAPGQRITISCTGSSSNIGAGYDVQWYQQVPGKAPKHLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYFCQSYDSSLSGYVVFGGGTKLTVL(SEQ ID NO:115) QVQLQESGPGLVKPSETLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHTGSPNYNPSLASRVTISMDKSKNQFSLNLRSVTAADTSVYYCARYGRGAFDIWGQGTMVTVSS(SEQ ID NO:116) >hFM114 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117) QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118) >hFM115 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM116 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:153) QVQLQESGPGLVKPSGTLSLTCDVSGGSISSNNWWSWVRQSPGKGLEWIGEIIHTGRTNYNPSLTSRVTILIDKSKNQFSLKLTSVTPEDTALYYCARLRGPFDIWGQGTMVTVSS(SEQ ID NO:154) >hFM117 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQQLPGTAPKLLIYADSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKLTVL(SEQ ID NO:117) QVQLVESGPGLVKPSGTLSLTCAVSGGSISSSDWWSWVRQPPGKGLEWIGEMRLSGTTNYNPSLKSRVAISMDKSKNHFSLNLSSVTAADTAVYYCANADYYTHHYWGQGTLVTVSS(SEQ ID NO:118) >hFM118 VLTQPPSVSGAPGQRVTISCTGGSTNIGAGYDVHWYQQLPGTAPKLLIYGNNNRPSGVPDRFSGSQSGASASLAITGLQADDEADYYCQSYDSRLDGSKVFGTGTKVTVL(SEQ ID NO:135) QVQLQESGPGLVKPSETLSLTCAVSGVSISSHNWWSWVRQTPGKGLEWIGEMSHSGIPNYNPSLESRVTISLDKSKNQFSLILRSVTAADTAMYYCVGGSGSYSYWGQGTLVTVSS(SEQ ID NO:136) >hFM120 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGFWVFGGGTKLTVL(SEQ ID NO:112) QVQLQESGPGLVKPSGTLSLTCAVSGGSISSSNWWSWVRQPPGKGLEWIGEISHSGSTNYNPSLKSRVTISVDNSKNQFSLQLSSVTAADTAVYYCVSNASGRRGFAWGQGTLVTVSS(SEQ ID NO:113) >hFM121 QSVLTQPPSVSAAPGQKVTISCSGSTSNVGNYYVAWYQKLPGEAPKVLIHDNNRRPSGIPVRFSGSKSGTSATLGITGLQTGDEAEYYCAVWDSSLNGYVFGGGTKLTVL(SEQ ID NO:128) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSLTAADTAVYYCARVNGGESDYWGQGTLVTVSS(SEQ ID NO:111) >hFM122 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM123 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLSVFGTGTKVTVL(SEQ ID NO:129) QLQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGIPNYNPSLASRVTISIDRSQNQFSLKLTSVTAADTAVYYCARGGGRFDFWGQGTLVTVSS(SEQ ID NO:130) >hFM124 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131) QVQLQQSGPGLVKPFGRPCPLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:133) >hFM125 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGANYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSSVFGGGTKLTVL(SEQ ID NO:131) QVQLQQSGPGLVKPSGTLSLTCDVSGGSISSRNWWTWVRQPPGKGLEWIGEIYHSGSTNYNPSLESRVAMSVDKSRYQFSLRLSSVTAADTAVYYCARRRDGYFDYWGQGTLVTVSS(SEQ ID NO:132) >hFM126 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNINRPSGVPDRFSGSKSGASASLAITGLQAEDEADYYCQSYDSSLSGLRVFGTGTKVTVL(SEQ ID NO:103) QVQLQESGPGLVEPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGSDMDVWGKGTMVTVSS(SEQ ID NO:104) >hFM127 QAVLTQPSSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNRNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSKVFGTGTKVTVL(SEQ ID NO:134) QLQLQESGPGLVKPSETLSLTCTVSGGSISSGNYWSWVRQSPEKGLEWIGEISHSGITNYNPSLQSRVTIALDKSKNHFSLNVNSVTAADTAVYYCVGTLRTWFDYWGQGTLVTVSS(SEQ ID NO:105) >hFM128 QAVLTQPSSVSGAPGQRVTISCTGSSTNIGAGFDVHWYQQLPGTAPKLLIYGDKNRPSGVPDRFSGSKSGTSAYLAITGLQAEDEADYYCQTYDSRLSGSKVFGGGTKVTVL(SEQ ID NO:106) QLQLQESGPGLVKPSGTLSLTCAVSGVSISSRNWWSWVRQTPGKGLEWIGEISHSGSTNYNPSLKSRVTISSVDTSKNQFSLKLSSVTAADTAVYYCARNAGDIWGQGTMVTVSS(SEQ ID NO:107) >hFM130 QSVLTQPPSVSAAPGQKVTISCSGSSSDIGNNFVSWYQQLPGTAPKRLIYDNSKRPSGIPERFSGSKSGTSATLGITGLQTGDEADYYCGAWDTSLSAYVFGTGTKVTVL(SEQ ID NO:108) QVQLQQWGPGLVKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISSVDTSKNQFSLKLSSVTAADTAVYYCARYSNYRHYYYGMDVWGQGTLVTVSS (SEQ ID NO: 109).

22. A method comprising administering the binding partner of claim 21 to an individual who has received an antibody-drug conjugate (ADC), thereby reducing off-target toxicity of the ADC.

23. A polynucleotide encoding the binding partner of claim 21.

24. A polynucleotide that hybridizes to the polynucleotide of claim 23.

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