ANTI-FOLATE RECEPTOR ANTIBODIES, COMPOSITIONS COMPRISING ANTI-FOLATE RECEPTOR ANTIBODIES AND METHODS OF MAKING AND USING ANTI-FOLATE RECEPTOR ANTIBODIES

Abstract

The present disclosure relates to antibodies that selectively bind to folate receptor alpha (FOLR1) and its isoforms and homologs, and compositions comprising the antibodies. Also provided are methods of using the antibodies, such as therapeutic and diagnostic methods.

Description

FIELD OF THE INVENTION

The present disclosure generally relates to antibodies with binding specificity for folate receptor alpha (FOLR1) and compositions comprising the antibodies, including pharmaceutical compositions, diagnostic compositions, and kits. Also provided are methods of making anti-folate receptor antibodies, and methods of using anti-folate receptor antibodies, for example, for therapeutic purposes, diagnostic purposes, and research purposes.

BACKGROUND

Folate receptors, or folate binding proteins (FBPs), include single chain glycoproteins that bind and contribute to the update of folates and other compounds in vivo. Elwood, 1989, J. Biol. Chem. 264:14893-14901. Certain folate receptors are single-chain glycoproteins with a high affinity binding site for folate and other compounds such as methotrexate. Elwood, p. 14893. The human FOLR1 gene encodes the adult folate receptor, a 30 kDa polypeptide with about 257 amino acids with three potential N-linked glycosylation sites. Elwood, p. 14893; Lacey et al., 1989, J. Clin. Invest. 84:715-720. Homologous genes and polypeptides have been identified in dozens of species.

The mature folate receptor glycoprotein has a size of about 42 kDa and has been observed to participate in the internalization of folates and antifolates into cells. Elwood et al., 1997, Biochemistry 36:1467-1478. Expression has been observed in human cerebellum and kidney cells, along with human cancer cell lines. Elwood et al., 1997, p. 1467. In addition to internalization of folate, a folate receptor has been shown to be a significant cofactor for cellular entry of viruses, particularly Marburg and Ebola viruses. Chan et al., 2001, Cell 106:117-126. Due to these internalization properties, the folate receptor has been proposed as a target for diagnostic and therapeutic agents. For instance, diagnostic and therapeutic agents have been linked to folate for internalization into cells expressing the folate receptor. See, e.g., Leamon, 2008, Curr. Opin. Investig. Drugs 9:1277-1286; Paulos et al., 2004, Adv. Drug Del. Rev. 56:1205-1217.

Folate receptor alpha (FolRα or FOLR1) is a glycosylphosphatidylinositol linked cell-surface glycoprotein that has high affinity for folates. Except for low levels in kidney and lung, most normal tissues do not express FOLR1, but high levels of FOLR1 have been found in serous and endometrioid epithelial ovarian cancer, endometrial adenocarcinoma, non-small cell lung carcinoma (NSCLC) of the adenocarcinoma subtype, and triple-negative breast cancer (TNBC). FOLR1 expression is maintained in metastatic foci and recurrent carcinomas in ovarian cancer patients, and FOLR1 expression has been observed after chemotherapy in epithelial ovarian and endometrial cancers. These properties, together with the highly restricted expression of FOLR1 on normal tissues, make FOLR1 a highly promising target for cancer therapy. As such, the folate receptor provides a potential target for diagnostics and therapeutics for cancers and inflammatory conditions. New antibodies are needed for specific binding and targeting of these folate receptors.

There is a need for improved methods of modulating the immune regulation of folate receptor alpha (FOLR1) and the downstream signaling processes activated by folate receptor alpha (FOLR1). Moreover, given the specific expression of folate receptor alpha (FOLR1) in cancer- and carcinoma-transformed cells and lower expression in non-cancer tissue, there is a need for improved therapeutics that can specifically target cells and tissues that overexpress folate receptor alpha (FOLR1).

SUMMARY

Provided herein are antibodies that selectively bind folate receptor alpha (FOLR1). In some embodiments, the antibodies bind human folate receptor alpha. In some embodiments, the antibodies also bind homologs of human folate receptor alpha. In some aspects, the homologs include a cynomolgus monkey homolog and mouse homolog.

In some embodiments, the antibodies comprise at least one CDR sequence defined by a consensus sequence provided in this disclosure. In some embodiments, the antibodies comprise an illustrative CDR, V_H, or V_L sequence provided in this disclosure, or a variant thereof. In some aspects, the variant is a variant with one or more conservative amino acid substitutions.

Also provided are compositions and kits comprising the antibodies. In some embodiments, the compositions are pharmaceutical compositions. Any suitable pharmaceutical composition may be used. In some embodiments, the pharmaceutical composition is a composition for parenteral administration.

This disclosure also provides methods of using the anti-folate receptor antibodies provided herein. In some embodiments, the method is a method of treatment. In some embodiments, the method is a diagnostic method. In some embodiments, the method is an analytical method. In some embodiments, the method is a method of purifying and/or quantifying folate receptor alpha (FOLR1).

In some embodiments, the antibodies are used to treat a disease or condition. In some aspects, the disease or condition is selected from a cancer, autoimmune disease, and infection.

These and other embodiments of the invention along with many of its features are described in more detail in conjunction with the text below and attached figures.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides a comparison of the Kabat and Chothia numbering systems for CDR-H1. Adapted from Martin A. C. R. (2010). Protein Sequence and Structure Analysis of Antibody Variable Domains. In R. Kontermann & S. Diibel (Eds.), Antibody Engineering vol. 2 (pp. 33-51). Springer-Verlag, Berlin Heidelberg.

FIGS. 2-4 provide alignments of the V_H sequences (SEQ ID NOs: 308-366) from the variant antibodies provided herein. CDRs according to Chothia are outlined/boxed, and CDRs according to Kabat are underlined.

FIG. 5 provides alignments of the V_L sequences (SEQ ID NOs: 367-369) from trastuzumab and the variant antibodies provided herein. CDRs according to Chothia are outlined/boxed, and CDRs according to Kabat are underlined.

DETAILED DESCRIPTION OF THE EMBODIMENTS

1. Definitions

Unless otherwise defined, all terms of art, notations and other scientific terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a difference over what is generally understood in the art. The techniques and procedures described or referenced herein are generally well understood and commonly employed using conventional methodologies by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 2nd ed. (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer-defined protocols and conditions unless otherwise noted.

As used herein, the singular forms “a,” “an,” and “the” include the plural referents unless the context clearly indicates otherwise.

The term “about” indicates and encompasses an indicated value and a range above and below that value. In certain embodiments, the term “about” indicates the designated value±10%, +5%, or +1%. In certain embodiments, the term “about” indicates the designated value±one standard deviation of that value.

The term “combinations thereof” includes every possible combination of elements to which the term refers to. For example, a sentence stating that “if” α₂ is A, then α₃ is not D; α₅ is not S; or α₆ is not S; or combinations thereof includes the following combinations when α₂ is A: (1) α₃ is not D; (2) α₅ is not S; (3) α₆ is not S; (4) α₃ is not D; α₅ is not S; and α₆ is not S; (5) α₃ is not D and α₅ is not S; (6) α₃ is not D and α₆ is not S; and (7) α₅ is not S and α₆ is not S.

The terms “folate receptor alpha” and “folate receptor 1” are used interchangeably herein. Folate receptor alpha is also known by synonyms, including FOLR1, FolRα, folate binding protein, FBP, adult folate binding protein, Folbp1, FR-alpha, FRα, KB cells FBP, and ovarian tumor-associated antigen MOv18, among others. Unless specified otherwise, the terms include any variants, isoforms and species homologs of human folate receptor alpha that are naturally expressed by cells, or that are expressed by cells transfected with a folate receptor alpha or FOLR1 gene. Folate receptor alpha proteins include, for example, human folate receptor alpha (SEQ ID NO: 1). In some embodiments, folate receptor alpha proteins include cynomolgus monkey folate receptor alpha (SEQ ID NO: 2). In some embodiments, folate receptor alpha proteins include murine folate receptor alpha (SEQ ID NO: 3).

The term “immunoglobulin” refers to a class of structurally related proteins generally comprising two pairs of polypeptide chains: one pair of light (L) chains and one pair of heavy (H) chains. In an “intact immunoglobulin,” all four of these chains are interconnected by disulfide bonds. The structure of immunoglobulins has been well characterized. See, e.g., Paul, Fundamental Immunology 7th ed., Ch. 5 (2013) Lippincott Williams & Wilkins, Philadelphia, Pa. Briefly, each heavy chain typically comprises a heavy chain variable region (VI) and a heavy chain constant region (C_H). The heavy chain constant region typically comprises three domains, abbreviated C_H1, C_H2, and C_H3. Each light chain typically comprises a light chain variable region (V_L) and a light chain constant region. The light chain constant region typically comprises one domain, abbreviated C_L.

The term “antibody” describes a type of immunoglobulin molecule and is used herein in its broadest sense. An antibody specifically includes intact antibodies (e.g., intact immunoglobulins), and antibody fragments. Antibodies comprise at least one antigen-binding domain. One example of an antigen-binding domain is an antigen binding domain formed by a V_H-V_L dimer. A “folate receptor alpha antibody,” “anti-folate receptor alpha antibody,” “folate receptor alpha Ab,” “folate receptor alpha-specific antibody,” “anti-folate receptor alpha Ab,” “FOLR1 antibody,” “FolRα antibody,” “anti-FOLR1 antibody,” “anti-FolRα antibody,” “FOLR1 Ab,” “FolRα Ab,” “FOLR1-specific antibody,” “FolRα-specific antibody,” “anti-FolRα Ab,” or “anti-FOLR1 Ab” is an antibody, as described herein, which binds specifically to folate receptor alpha or FOLR1. In some embodiments, the antibody binds the extracellular domain of folate receptor alpha (FOLR1).

The V_H and V_L regions may be further subdivided into regions of hypervariability (“hypervariable regions (HVRs);” also called “complementarity determining regions” (CDRs)) interspersed with regions that are more conserved. The more conserved regions are called framework regions (FRs). Each V_H and V_L generally comprises three CDRs and four FRs, arranged in the following order (from N-terminus to C-terminus): FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. The CDRs are involved in antigen binding, and influence antigen specificity and binding affinity of the antibody. See Kabat et al., Sequences of Proteins of Immunological Interest 5th ed. (1991) Public Health Service, National Institutes of Health, Bethesda, Md., incorporated by reference in its entirety.

The light chain from any vertebrate species can be assigned to one of two types, called kappa and lambda, based on the sequence of the constant domain.

The heavy chain from any vertebrate species can be assigned to one of five different classes (or isotypes): IgA, IgD, IgE, IgG, and IgM. These classes are also designated α, δ, ε, γ, and μ, respectively. The IgG and IgA classes are further divided into subclasses on the basis of differences in sequence and function. Humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

The amino acid sequence boundaries of a CDR can be determined by one of skill in the art using any of a number of known numbering schemes, including those described by Kabat et al., supra (“Kabat” numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 (“Chothia” numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732-745 (“Contact” numbering scheme); Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 (“IMGT” numbering scheme); and Honegge and Plückthun, J. Mol. Biol., 2001, 309:657-70 (“AHo” numbering scheme), each of which is incorporated by reference in its entirety.

Table 1 provides the positions of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 as identified by the Kabat and Chothia schemes. For CDR-H1, residue numbering is provided using both the Kabat and Chothia numbering schemes.

TABLE 1
Residues in CDRs according to Kabat and Chothia
numbering schemes.
	CDR	Kabat	Chothia
	L1	L24-L34	L24-L34
	L2	L50-L56	L50-L56
	L3	L89-L97	L89-L97
	H1 (Kabat Numbering)	H31-H35B	H26-H32 or H34*
	H1 (Chothia Numbering)	H31-H35	H26-H32
	H2	H50-H65	H52-H56
	H3	H95-H102	H95-H102
	*The C-terminus of CDR-H1, when numbered using the Kabat numbering convention, varies between H32 and H34, depending on the length of the CDR, as illustrated in FIG. 1.

Unless otherwise specified, the numbering scheme used for identification of a particular CDR herein is the Kabat/Chothia numbering scheme. Where the residues encompassed by these two numbering schemes diverge (e.g., CDR-H1 and/or CDR-H2), the numbering scheme is specified as either Kabat or Chothia. For convenience, CDR-H3 is sometimes referred to herein as either Kabat or Chothia. However, this is not intended to imply differences in sequence where they do not exist, and one of skill in the art can readily confirm whether the sequences are the same or different by examining the sequences.

CDRs may be assigned, for example, using antibody numbering software, such as Abnum, available at http://www.bioinf.org.uk/abs/abnum/, and described in Abhinandan and Martin, Immunology, 2008, 45:3832-3839, incorporated by reference in its entirety.

The “EU numbering scheme” is generally used when referring to a residue in an antibody heavy chain constant region (e.g., as reported in Kabat et al., supra). Unless stated otherwise, the EU numbering scheme is used to refer to residues in antibody heavy chain constant regions described herein.

An “antibody fragment” comprises a portion of an intact antibody, such as the antigen binding or variable region of an intact antibody. Antibody fragments include, for example, Fv fragments, Fab fragments, F(ab′)₂ fragments, Fab′ fragments, scFv (sFv) fragments, and scFv-Fc fragments.

“Fv” fragments comprise a non-covalently-linked dimer of one heavy chain variable domain and one light chain variable domain.

“Fab” fragments comprise, in addition to the heavy and light chain variable domains, the constant domain of the light chain and the first constant domain (C_H1) of the heavy chain. Fab fragments may be generated, for example, by recombinant methods or by papain digestion of a full-length antibody.

“F(ab′)₂” fragments contain two Fab′ fragments joined, near the hinge region, by disulfide bonds. F(ab′)₂ fragments may be generated, for example, by recombinant methods or by pepsin digestion of an intact antibody. The F(ab′) fragments can be dissociated, for example, by treatment with β-mercaptoethanol.

“Single-chain Fv” or “sFv” or “scFv” antibody fragments comprise a V_H domain and a V_L domain in a single polypeptide chain. The V_H and V_L are generally linked by a peptide linker. See Plückthun A. (1994). In some embodiments, the linker is SEQ ID NO: 377. In some embodiments, the linker is SEQ ID NO: 378. Antibodies from Escherichia coli. In Rosenberg M. & Moore G. P. (Eds.), The Pharmacology of Monoclonal Antibodies vol. 113 (pp. 269-315). Springer-Verlag, New York, incorporated by reference in its entirety.

“scFv-Fc” fragments comprise an scFv attached to an Fc domain. For example, an Fc domain may be attached to the C-terminus of the scFv. The Fc domain may follow the V_H or V_L, depending on the orientation of the variable domains in the scFv (i.e., V_H-V_L or V_L-V_H). Any suitable Fc domain known in the art or described herein may be used. In some cases, the Fc domain comprises an IgG1 Fc domain. In some embodiments, the IgG1 Fc domain comprises SEQ ID NO: 370, or a portion thereof. SEQ ID NO: 370 provides the sequence of C_H1, C_H2, and C_H3 of the human IgG1 constant region.

The term “monoclonal antibody” refers to an antibody from a population of substantially homogeneous antibodies. A population of substantially homogeneous antibodies comprises antibodies that are substantially similar and that bind the same epitope(s), except for variants that may normally arise during production of the monoclonal antibody. Such variants are generally present in only minor amounts. A monoclonal antibody is typically obtained by a process that includes the selection of a single antibody from a plurality of antibodies. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, yeast clones, bacterial clones, or other recombinant DNA clones. The selected antibody can be further altered, for example, to improve affinity for the target (“affinity maturation”), to humanize the antibody, to improve its production in cell culture, and/or to reduce its immunogenicity in a subject.

The term “chimeric antibody” refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.

“Humanized” forms of non-human antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. A humanized antibody is generally a human immunoglobulin (recipient antibody) in which residues from one or more CDRs are replaced by residues from one or more CDRs of a non-human antibody (donor antibody). The donor antibody can be any suitable non-human antibody, such as a mouse, rat, rabbit, chicken, or non-human primate antibody having a desired specificity, affinity, or biological effect. In some instances, selected framework region residues of the recipient antibody are replaced by the corresponding framework region residues from the donor antibody. Humanized antibodies may also comprise residues that are not found in either the recipient antibody or the donor antibody. Such modifications may be made to further refine antibody function. For further details, see Jones et al., Nature, 1986, 321:522-525; Riechmann et al., Nature, 1988, 332:323-329; and Presta, Curr. Op. Struct. Biol., 1992, 2:593-596, each of which is incorporated by reference in its entirety.

A “human antibody” is one which possesses an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or derived from a non-human source that utilizes a human antibody repertoire or human antibody-encoding sequences (e.g., obtained from human sources or designed de novo). Human antibodies specifically exclude humanized antibodies.

An “isolated antibody” is one that has been separated and/or recovered from a component of its natural environment. Components of the natural environment may include enzymes, hormones, and other proteinaceous or nonproteinaceous materials. In some embodiments, an isolated antibody is purified to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence, for example by use of a spinning cup sequenator. In some embodiments, an isolated antibody is purified to homogeneity by gel electrophoresis (e.g., SDS-PAGE) under reducing or nonreducing conditions, with detection by Coomassie blue or silver stain. An isolated antibody includes an antibody in situ within recombinant cells, since at least one component of the antibody's natural environment is not present. In some aspects, an isolated antibody is prepared by at least one purification step.

In some embodiments, an isolated antibody is purified to at least 80%, 85%, 90%, 95%, or 99% by weight. In some embodiments, an isolated antibody is purified to at least 80%, 85%, 90%, 95%, or 99% by volume. In some embodiments, an isolated antibody is provided as a solution comprising at least 85%, 90%, 95%, 98%, 99% to 100% by weight. In some embodiments, an isolated antibody is provided as a solution comprising at least 85%, 90%, 95%, 98%, 99% to 100% by volume.

“Affinity” refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of a molecule X for its partner Y can be represented by the dissociation constant (K_D). Affinity can be measured by common methods known in the art, including those described herein. Affinity can be determined, for example, using surface plasmon resonance (SPR) technology, such as a Biacore® instrument. In some embodiments, the affinity is determined at 25° C.

With regard to the binding of an antibody to a target molecule, the terms “specific binding,” “specifically binds to,” “specific for,” “selectively binds,” and “selective for” a particular antigen (e.g., a polypeptide target) or an epitope on a particular antigen mean binding that is measurably different from a non-specific or non-selective interaction. Specific binding can be measured, for example, by determining binding of a molecule compared to binding of a control molecule. Specific binding can also be determined by competition with a control molecule that mimics the antibody binding site on the target. In that case, specific binding is indicated if the binding of the antibody to the target is competitively inhibited by the control molecule.

The term “k_d” (sec⁻¹), as used herein, refers to the dissociation rate constant of a particular antibody-antigen interaction. This value is also referred to as the k_off value.

The term “k_a” (M⁻¹×sec⁻¹), as used herein, refers to the association rate constant of a particular antibody-antigen interaction. This value is also referred to as the k_on value.

The term “K_D” (M), as used herein, refers to the dissociation equilibrium constant of a particular antibody-antigen interaction. K_D=k_d/k_a.

The term “K_A” (M⁻¹), as used herein, refers to the association equilibrium constant of a particular antibody-antigen interaction. K_A=k_a/k_d.

An “affinity matured” antibody is one with one or more alterations in one or more CDRs or FRs that result in an improvement in the affinity of the antibody for its antigen, compared to a parent antibody which does not possess the alteration(s). In one embodiment, an affinity matured antibody has nanomolar or picomolar affinity for the target antigen. Affinity matured antibodies may be produced using a variety of methods known in the art. For example, Marks et al. (Bio/Technology, 1992, 10:779-783, incorporated by reference in its entirety) describes affinity maturation by V_H and V_L domain shuffling. Random mutagenesis of CDR and/or framework residues is described by, for example, Barbas et al. (Proc. Nat. Acad. Sci. U.S.A., 1994, 91:3809-3813); Schier et al., Gene, 1995, 169:147-155; Yelton et al., J. Immunol., 1995, 155:1994-2004; Jackson et al., J. Immunol., 1995, 154:3310-33199; and Hawkins et al, J. Mol. Biol., 1992, 226:889-896, each of which is incorporated by reference in its entirety.

When used herein in the context of two or more antibodies, the term “competes with” or “cross-competes with” indicates that the two or more antibodies compete for binding to an antigen (e.g., folate receptor alpha, or FOLR1). In one exemplary assay, FOLR1 is coated on a plate and allowed to bind a first antibody, after which a second, labeled antibody is added. If the presence of the first antibody reduces binding of the second antibody, then the antibodies compete. In another exemplary assay, a first antibody is coated on a plate and allowed to bind the antigen, and then the second antibody is added. The term “competes with” also includes combinations of antibodies where one antibody reduces binding of another antibody, but where no competition is observed when the antibodies are added in the reverse order. However, in some embodiments, the first and second antibodies inhibit binding of each other, regardless of the order in which they are added. In some embodiments, one antibody reduces binding of another antibody to its antigen by at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%.

The term “epitope” means a portion of an antigen capable of specific binding to an antibody. Epitopes frequently consist of surface-accessible amino acid residues and/or sugar side chains and may have specific three dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents. An epitope may comprise amino acid residues that are directly involved in the binding, and other amino acid residues, which are not directly involved in the binding. The epitope to which an antibody binds can be determined using known techniques for epitope determination such as, for example, testing for antibody binding to variants of folate receptor alpha (FOLR1) with different point-mutations.

Percent “identity” between a polypeptide sequence and a reference sequence, is defined as the percentage of amino acid residues in the polypeptide sequence that are identical to the amino acid residues in the reference sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

A “conservative substitution” or a “conservative amino acid substitution,” refers to the substitution of an amino acid with a chemically or functionally similar amino acid. Conservative substitution tables providing similar amino acids are well known in the art. Polypeptide sequences having such substitutions are known as “conservatively modified variants.” Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles. By way of example, the groups of amino acids provided in Tables 2-4 are, in some embodiments, considered conservative substitutions for one another.

TABLE 2
Selected groups of amino acids that are considered conservative
substitutions for one another, in certain embodiments.
Acidic Residues                  D and E
Basic Residues                   K, R, and H
Hydrophilic Uncharged Residues   S, T, N, and Q
Aliphatic Uncharged Residues     G, A, V, L, and I
Non-polar Uncharged Residues     C, M, and P
Aromatic Residues                F, Y, and W
Alcohol Group-Containing Residues S and T
Aliphatic Residues               I, L, V, and M
Cycloalkenyl-associated Residues F, H, W, and Y
Hydrophobic Residues             A, C, F, G, H, I, L, M, R, T, V,
                                 W, and Y
Negatively Charged Residues      D and E
Polar Residues                   C, D, E, H, K, N, Q, R, S, and T
Positively Charged Residues      H, K, and R
Small Residues                   A, C, D, G, N, P, S, T, and V
Very Small Residues              A, G, and S
Residues Involved in Turn Formation A, C, D, E, G, H, K, N, Q, R, S,
                                 P, and T
Flexible Residues                Q, T, K, S, G, P, D, E, and R

TABLE 3
Additional selected groups of amino acids that are considered
conservative substitutions for one another, in certain embodiments.
Group 1 A, S, and T
Group 2 D and E
Group 3 N and Q
Group 4 R and K
Group 5 I, L, and M
Group 6 F, Y, and W

TABLE 4
Further selected groups of amino acids that are considered
conservative substitutions for one another, in certain embodiments.
Group A A and G
Group B D and E
Group C N and Q
Group D R, K, and H
Group E I, L, M, V
Group F F, Y, and W
Group G S and T
Group H C and M

Additional conservative substitutions may be found, for example, in Creighton, Proteins: Structures and Molecular Properties 2nd ed. (1993) W. H. Freeman & Co., New York, N.Y. An antibody generated by making one or more conservative substitutions of amino acid residues in a parent antibody is referred to as a “conservatively modified variant.”

The term “amino acid” refers to the twenty common naturally occurring amino acids. Naturally occurring amino acids include alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine (Cys; C); glutamic acid (Glu; E), glutamine (Gln; Q), Glycine (Gly; G); histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Val; V).

“Treating” or “treatment” of any disease or disorder refers, in certain embodiments, to ameliorating a disease or disorder that exists in a subject. In another embodiment, “treating” or “treatment” includes ameliorating at least one physical parameter, which may be indiscernible by the subject. In yet another embodiment, “treating” or “treatment” includes modulating the disease or disorder, either physically (e.g., stabilization of a discernible symptom) or physiologically (e.g., stabilization of a physical parameter) or both. In yet another embodiment, “treating” or “treatment” includes delaying or preventing the onset of the disease or disorder.

As used herein, the term “therapeutically effective amount” or “effective amount” refers to an amount of an antibody or composition that when administered to a subject is effective to treat a disease or disorder. In some embodiments, a therapeutically effective amount or effective amount refers to an amount of an antibody or composition that when administered to a subject is effective to prevent or ameliorate a disease or the progression of the disease, or result in amelioration of symptoms.

As used herein, the term “subject” means a mammalian subject. Exemplary subjects include, but are not limited to humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, avians, goats, and sheep. In certain embodiments, the subject is a human. In some embodiments, the subject has a disease that can be treated or diagnosed with an antibody provided herein. In some embodiments, the disease is gastric carcinoma, colorectal carcinoma, renal cell carcinoma, cervical carcinoma, non-small cell lung carcinoma, ovarian cancer, breast cancer, triple-negative breast cancer, endometrial cancer, prostate cancer, and/or a cancer of epithelial origin.

2. Antibodies

Provided herein are antibodies that selectively bind human folate receptor alpha. In some aspects, the antibody selectively binds to the extracellular domain of human folate receptor alpha (human FOLR1).

In some embodiments, the antibody binds to a homolog of human FOLR1. In some aspects, the antibody binds to a homolog of human FOLR1 from a species selected from monkeys, mice, dogs, cats, rats, cows, horses, goats and sheep. In some aspects, the homolog is a cynomolgus monkey homolog. In some aspects, the homolog is a mouse or murine analog.

In some embodiments, the antibodies comprise at least one CDR sequence defined by a consensus sequence provided in this disclosure. In some embodiments, the antibodies comprise an illustrative CDR, V_H, or V_L sequence provided in this disclosure, or a variant thereof. In some aspects, the variant is a variant with a conservative amino acid substitution.

In some embodiments, the antibody has one or more CDRs having particular lengths, in terms of the number of amino acid residues. In some embodiments, the Chothia CDR-H1 of the antibody is 6, 7, or 8 residues in length. In some embodiments, the Kabat CDR-H1 of the antibody is 4, 5, or 6 residues in length. In some embodiments, the Chothia CDR-H2 of the antibody is 5, 6, or 7 residues in length. In some embodiments, the Kabat CDR-H2 of the antibody is 16, 17, or 18 residues in length. In some embodiments, the Kabat/Chothia CDR-H3 of the antibody is 13, 14, 15, 16, or 17 residues in length.

In some aspects, the Kabat/Chothia CDR-L1 of the antibody is 11, 12, 13, 14, 15, 16, 17, or 18 residues in length. In some aspects, the Kabat/Chothia CDR-L2 of the antibody is 6, 7, or 8 residues in length. In some aspects, the Kabat/Chothia CDR-L3 of the antibody is 8, 9, or 10 residues in length.

In some embodiments, the antibody comprises a light chain. In some aspects, the light chain is a kappa light chain. In some aspects, the light chain is a lambda light chain.

In some embodiments, the antibody comprises a heavy chain. In some aspects, the heavy chain is an IgA. In some aspects, the heavy chain is an IgD. In some aspects, the heavy chain is an IgE. In some aspects, the heavy chain is an IgG. In some aspects, the heavy chain is an IgM. In some aspects, the heavy chain is an IgG1. In some aspects, the heavy chain is an IgG2. In some aspects, the heavy chain is an IgG3. In some aspects, the heavy chain is an IgG4. In some aspects, the heavy chain is an IgA1. In some aspects, the heavy chain is an IgA2.

In some embodiments, the antibody is an antibody fragment. In some aspects, the antibody fragment is an Fv fragment. In some aspects, the antibody fragment is a Fab fragment. In some aspects, the antibody fragment is a F(ab′)₂ fragment. In some aspects, the antibody fragment is a Fab′ fragment. In some aspects, the antibody fragment is an scFv (sFv) fragment. In some aspects, the antibody fragment is an scFv-Fc fragment.

In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a polyclonal antibody.

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

In some embodiments, the antibody is an affinity matured antibody. In some aspects, the antibody is an affinity matured antibody derived from an illustrative sequence provided in this disclosure.

The antibodies provided herein may be useful for the treatment of a variety of diseases and conditions including cancers. In some embodiments, the antibodies provided herein may be useful for the treatment of cancers of solid tumors. For example, the antibodies provided herein can be useful for the treatment of colorectal cancer.

2.1 CDR-H3 Sequences

In some embodiments, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of a CDR-H3 sequence of an illustrative antibody or V_H sequence provided herein. In some aspects, the CDR-H3 sequence is a CDR-H3 sequence of a V_H sequence provided in SEQ ID NOs.: 308-366.

In some embodiments, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs.: 240-298. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 240. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 241. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 242. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 243. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 244. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 245. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 246. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 247. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 248. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 249. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 250. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 251. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 252. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 253. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 254. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 255. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 256. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 257. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 258. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 259. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 260. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 261. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 262. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 263. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 264. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 265. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 266. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 267. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 268. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 269. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 270. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 271. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 272. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 273. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 274. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 275. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 276. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 277. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 278. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 279. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 280. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 281. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 282. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 283. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 284. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 285. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 286. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 287. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 288. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 289. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 290. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 291. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 292. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 293. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 294. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 295. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 296. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 297. In some aspects, the antibody comprises a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 298.

In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-H3 sequence provided in this disclosure. In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-H3 sequences provided in this disclosure. In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-H3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.2 V_H Sequences Comprising Illustrative CDRs

In some embodiments, the antibody comprises a V_H sequence comprising one or more CDR-H sequences comprising, consisting of, or consisting essentially of one or more illustrative CDR-H sequences provided in this disclosure, and variants thereof. In some embodiments, the CDR-H sequences comprise, consist of, or consist essentially of one or more CDR-H sequences provided in a V_H sequence selected from SEQ ID NOs: 308-366.

2.2.1. V_H Sequences Comprising Illustrative Kabat CDRs

In some embodiments, the antibody comprises a V_H sequence comprising one or more Kabat CDR-H sequences comprising, consisting of, or consisting essentially of one or more illustrative Kabat CDR-H sequences provided in this disclosure, and variants thereof.

2.2.1.1. Kabat CDR-H3

In some embodiments, the antibody comprises a V_H sequence comprising a CDR-H3 sequence, wherein the CDR-H3 sequence comprises, consists of, or consists essentially of a Kabat CDR-H3 sequence of an illustrative antibody or V_H sequence provided herein. In some aspects, the Kabat CDR-H3 sequence is a Kabat CDR-H3 sequence of a V_H sequence provided in SEQ ID NOs: 308-366.

In some embodiments, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs.: 240-298. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 240. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 241. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 242. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 243. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 244. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 245. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 246. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 247. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 248. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 249. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 250. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 251. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 252. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 253. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 254. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 255. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 256. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 257. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 258. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 259. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 260. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 261. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 262. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 263. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 264. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 265. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 266. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 267. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 268. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 269. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 270. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 271. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 272. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 273. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 274. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 275. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 276. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 277. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 278. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 279. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 280. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 281. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 282. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 283. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 284. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 285. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 286. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 287. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 288. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 289. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 290. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 291. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 292. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 293. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 294. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 295. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 296. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 297. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 298.

2.2.1.2. Kabat CDR-H2

In some embodiments, the antibody comprises a V_H sequence comprising a CDR-H2 sequence, wherein the CDR-H2 sequence comprises, consists of, or consists essentially of a Kabat CDR-H2 sequence of an illustrative antibody or V_H sequence provided herein. In some aspects, the Kabat CDR-H2 sequence is a Kabat CDR-H2 sequence of a V_H sequence provided in SEQ ID NOs: 308-366.

In some embodiments, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 181-239. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 181. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 182. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 183. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 184. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 185. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 186. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 187. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 188. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 189. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 190. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 191. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 192. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 193. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 194. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 195. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 196. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 197. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 198. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 199. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 200. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 201. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 202. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 203. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 204. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 205. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 206. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 207. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 208. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 209. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 210. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 211. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 212. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 213. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 214. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 215. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 216. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 217. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 218. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 219. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 220. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 221. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 222. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 223. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 224. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 225. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 226. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 227. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 228. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 229. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 230. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 231. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 232. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 233. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 234. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 235. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 236. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 237. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 238. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 239.

2.2.1.3. Kabat CDR-H1

In some embodiments, the antibody comprises a V_H sequence comprising a CDR-H1 sequence, wherein the CDR-H1 sequence comprises, consists of, or consists essentially of a Kabat CDR-H1 sequence of an illustrative antibody or V_H sequence provided herein. In some aspects, the Kabat CDR-H1 sequence is a Kabat CDR-H1 sequence of a V_H sequence provided in SEQ ID NOs: 308-366.

In some embodiments, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 63-121. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 63. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 64. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 65. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 66. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 67. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 68. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 69. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 70. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 71. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 72. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 73. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 74. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 75. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 76. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 77. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 78. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 79. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 80. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 81. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 82. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 83. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 84. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 85. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 86. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 87. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 88. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 89. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 90. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 91. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 92. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 93. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 94. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 95. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 96. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 97. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 98. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 99. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 100. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 101. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 102. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 103. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 104. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 105. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 106. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 107. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 108. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 109. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 110. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 111. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 112. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 113. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 114. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 115. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 116. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 117. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 118. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 119. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 120. In some aspects, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 121.

2.2.1.4. Kabat CDR-H3+Kabat CDR-H2

In some embodiments, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 240-298, and a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 181-239. In some aspects, the Kabat CDR-H3 sequence and the Kabat CDR-H2 sequence are both from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Kabat CDR-H3 and Kabat CDR-H2 are both from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.1.5. Kabat CDR-H3+Kabat CDR-H1

In some embodiments, the antibody comprises a V_H sequence comprising a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 240-298, and a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 63-121. In some aspects, the Kabat CDR-H3 sequence and the Kabat CDR-H1 sequence are both from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Kabat CDR-H3 and Kabat CDR-H1 are both from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.1.6. Kabat CDR-H1+Kabat CDR-H2

In some embodiments, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 63-121 and a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 181-239. In some aspects, the Kabat CDR-H1 sequence and the Kabat CDR-H2 sequence are both from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Kabat CDR-H1 and Kabat CDR-H2 are both from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.1.7. Kabat CDR-H1+Kabat CDR-H2+Kabat CDR-H3

In some embodiments, the antibody comprises a V_H sequence comprising a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 63-121, a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 181-239, and a Kabat CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 240-298. In some aspects, the Kabat CDR-H1 sequence, Kabat CDR-H2 sequence, and Kabat CDR-H3 sequence are all from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Kabat CDR-H1, Kabat CDR-H2, and Kabat CDR-H3 are all from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.1.8. Variants of V_H Sequences Comprising Illustrative Kabat CDRs

In some embodiments, the V_H sequences provided herein comprise a variant of an illustrative Kabat CDR-H3, CDR-H2, and/or CDR-H1 sequence provided in this disclosure.

In some aspects, the Kabat CDR-H3 sequence comprises, consists of, or consists essentially of a variant of an illustrative Kabat CDR-H3 sequence provided in this disclosure. In some aspects, the Kabat CDR-H3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Kabat CDR-H3 sequences provided in this disclosure. In some aspects, the Kabat CDR-H3 sequence comprises, consists of, or consists essentially of any of the illustrative Kabat CDR-H3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the Kabat CDR-H2 sequence comprises, consists of, or consists essentially of a variant of an illustrative Kabat CDR-H2 sequence provided in this disclosure. In some aspects, the Kabat CDR-H2 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Kabat CDR-H2 sequences provided in this disclosure. In some aspects, the Kabat CDR-H2 sequence comprises, consists of, or consists essentially of any of the illustrative Kabat CDR-H2 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the Kabat CDR-H1 sequence comprises, consists of, or consists essentially of a variant of an illustrative Kabat CDR-H1 sequence provided in this disclosure. In some aspects, the Kabat CDR-H1 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Kabat CDR-H1 sequences provided in this disclosure. In some aspects, the Kabat CDR-H1 sequence comprises, consists of, or consists essentially of any of the illustrative Kabat CDR-H1 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.2.2. V_H Sequences Comprising Illustrative Chothia CDRs

In some embodiments, the antibody comprises a V_H sequence comprising one or more Chothia CDR-H sequences comprising, consisting of, or consisting essentially of one or more illustrative Chothia CDR-H sequences provided in this disclosure, and variants thereof.

2.2.2.1. Chotia CDR-H3

In some embodiments, the antibody comprises a V_H sequence comprising a CDR-H3 sequence, wherein the CDR-H3 sequence comprises, consists of, or consists essentially of a Chothia CDR-H3 sequence of an illustrative antibody or V_H sequence provided herein. In some aspects, the Chothia CDR-H3 sequence is a Chothia CDR-H3 sequence of a V_H sequence provided in SEQ ID NOs: 308-366.

In some embodiments, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 240-298. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 240. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 241. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 242. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 243. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 244. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 245. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 246. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 247. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 248. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 249. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 250. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 251. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 252. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 253. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 254. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 255. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 256. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 257. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 258. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 259. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 260. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 261. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 262. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 263. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 264. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 265. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 266. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 267. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 268. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 269. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 270. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 271. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 272. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 273. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 274. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 275. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 276. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 277. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 278. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 279. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 280. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 281. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 282. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 283. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 284. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 285. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 286. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 287. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 288. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 289. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 290. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 291. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 292. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 293. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 294. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 295. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 296. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 297. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 298.

2.2.2.2. Chothia CDR-H2

In some embodiments, the antibody comprises a V_H sequence comprising a CDR-H2 sequence, wherein the CDR-H2 sequence comprises, consists of, or consists essentially of a Chothia CDR-H2 sequence of an illustrative antibody or V_H sequence provided herein. In some aspects, the Chothia CDR-H2 sequence is a Chothia CDR-H2 sequence of a V_H sequence provided in SEQ ID NOs: 308-366.

In some embodiments, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 122-180. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 122. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 123. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 124. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 125. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 126. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 127. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 128. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 129. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 130. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 131. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 132. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 133. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 134. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 135. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 136. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 137. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 138. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 139. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 140. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 141. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 142. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 143. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 144. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 145. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 146. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 147. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 148. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 149. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 150. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 151. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 152. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 153. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 154. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 155. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 156. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 157. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 158. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 159. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 160. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 161. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 162. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 163. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 164. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 165. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 166. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 167. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 168. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 169. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 170. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 171. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 172. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 173. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 174. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 175. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 176. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 177. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 178. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 179. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 180.

2.2.2.3. Chothia CDR-H1

In some embodiments, the antibody comprises a V_H sequence comprising a CDR-H1 sequence, wherein the CDR-H1 sequence comprises, consists of, or consists essentially of a Chothia CDR-H1 sequence of an illustrative antibody or V_H sequence provided herein. In some aspects, the Chothia CDR-H1 sequence is a Chothia CDR-H1 sequence of a V_H sequence provided in SEQ ID NOs: 308-366.

In some embodiments, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 4-62. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 4. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 5. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 6. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 7. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 8. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 9. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 10. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 11. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 12. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 13. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 14. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 15. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 16. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 17. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 18. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 19. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 20. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 21. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 22. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 23. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 24. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 25. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 26. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 27. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 28. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 29. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 30. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 31. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 32. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 33. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 34. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 35. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 36. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 37. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 38. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 39. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 40. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 41. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 42. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 43. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 44. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 45. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 46. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 47. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 48. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 49. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 50. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 51. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 52. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 53. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 54. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 55. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 56. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 57. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 58. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 59. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 60. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 61. In some aspects, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 62.

2.2.2.4. Chothia CDR-H3+Chothia CDR-H2

In some embodiments, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 240-298, and a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 122-180. In some aspects, the Chothia CDR-H3 sequence and the Chothia CDR-H2 sequence are both from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Chothia CDR-H3 and Chothia CDR-H2 are both from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.2.5. Chothia CDR-H3+Chothia CDR-H1

In some embodiments, the antibody comprises a V_H sequence comprising a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 240-298, and a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 4-62. In some aspects, the Chothia CDR-H3 sequence and the Chothia CDR-H1 sequence are both from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Chothia CDR-H3 and Chothia CDR-H1 are both from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.2.6. Chothia CDR-H1+Chothia CDR-H2

In some embodiments, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 4-62 and a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 122-180. In some aspects, the Chothia CDR-H1 sequence and the Chothia CDR-H2 sequence are both from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Chothia CDR-H1 and Chothia CDR-H2 are both from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.2.7. Chothia CDR-H1+Chothia CDR-H2+Chothia CDR-H3

In some embodiments, the antibody comprises a V_H sequence comprising a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 4-62, a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 122-180, and a Chothia CDR-H3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 240-298. In some aspects, the Chothia CDR-H1 sequence, Chothia CDR-H2 sequence, and Chothia CDR-H3 sequence are all from a single illustrative V_H sequence provided in this disclosure. For example, in some aspects, the Chothia CDR-H1, Chothia CDR-H2, and Chothia CDR-H3 are all from a single illustrative V_H sequence selected from SEQ ID NOs: 308-366.

2.2.2.8. Variants of V_H Sequences Comprising Illustrative Chothia CDRs

In some embodiments, the V_H sequences provided herein comprise a variant of an illustrative Chothia CDR-H3, CDR-H2, and/or CDR-H1 sequence provided in this disclosure.

In some aspects, the Chothia CDR-H3 sequence comprises, consists of, or consists essentially of a variant of an illustrative Chothia CDR-H3 sequence provided in this disclosure. In some aspects, the Chothia CDR-H3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Chothia CDR-H3 sequences provided in this disclosure. In some aspects, the Chothia CDR-H3 sequence comprises, consists of, or consists essentially of any of the illustrative Chothia CDR-H3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the Chothia CDR-H2 sequence comprises, consists of, or consists essentially of a variant of an illustrative Chothia CDR-H2 sequence provided in this disclosure. In some aspects, the Chothia CDR-H2 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Chothia CDR-H2 sequences provided in this disclosure. In some aspects, the Chothia CDR-H2 sequence comprises, consists of, or consists essentially of any of the illustrative Chothia CDR-H2 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the Chothia CDR-H1 sequence comprises, consists of, or consists essentially of a variant of an illustrative Chothia CDR-H1 sequence provided in this disclosure. In some aspects, the Chothia CDR-H1 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Chothia CDR-H1 sequences provided in this disclosure. In some aspects, the Chothia CDR-H1 sequence comprises, consists of, or consists essentially of any of the illustrative Chothia CDR-H1 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.3. V_H Sequences

In some embodiments, the antibody comprises, consists of, or consists essentially of a V_H sequence provided in SEQ ID NOs: 308-366.

In some embodiments, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 308-366. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 308. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 309. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 310. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 311. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 312. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 313. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 314. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 315. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 316. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 317. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 318. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 319. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 320. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 321. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 322. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 323. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 324. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 325. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 326. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 327. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 328. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 329. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 330. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 331. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 332. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 333. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 334. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 335. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 336. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 337. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 338. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 339. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 340. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 341. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 342. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 343. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 344. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 345. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 346. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 347. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 348. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 349. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 350. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 351. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 352. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 353. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 354. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 355. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 356. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 357. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 358. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 359. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 360. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 361. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 362. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 363. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 364. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 365. In some aspects, the antibody comprises a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 366.

2.3.1. Variants of V_H Sequences

In some embodiments, the V_H sequences provided herein comprise, consist of, or consist essentially of a variant of an illustrative V_H sequence provided in this disclosure.

In some aspects, the V_H sequence comprises, consists of, or consists essentially of a variant of an illustrative V_H sequence provided in this disclosure. In some aspects, the V_H sequence comprises, consists of, or consists essentially of a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with any of the illustrative V_H sequences provided in this disclosure.

In some embodiments, the V_H sequence comprises, consists of, or consists essentially of any of the illustrative V_H sequences provided in this disclosure having 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer, 16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 or fewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.4. CDR-L3 Sequences

In some embodiments, the antibody comprises a CDR-L3 sequence comprising, consisting of, or consisting essentially of a CDR-L3 sequence of an illustrative antibody or V_L sequence provided herein. In some aspects, the CDR-L3 sequence is a CDR-L3 sequence of a V_L sequence provided in SEQ ID NOs.: 367-369.

In some embodiments, the antibody comprises a CDR-L3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 305-307. In some aspects, the antibody comprises a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 305. In some aspects, the antibody comprises a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 306. In some aspects, the antibody comprises a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 307.

In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L3 sequence provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L3 sequences provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.5. V_L Sequences Comprising Illustrative CDRs

In some embodiments, the antibody comprises a V_L sequence comprising one or more CDR-L sequences comprising, consisting of, or consisting essentially of one or more illustrative CDR-L sequences provided in this disclosure, and variants thereof.

2.5.1. CDR-L3

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L3 sequence, wherein the CDR-L3 sequence comprises, consists of, or consists essentially of a CDR-L3 sequence of an illustrative antibody or V_L sequence provided herein. In some aspects, the CDR-L3 sequence is a CDR-L3 sequence of a V_L sequence provided in SEQ ID NOs.: 367-369.

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 305-307. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 305. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 306. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 307.

2.5.2. CDR-L2

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L2 sequence, wherein the CDR-L2 sequence comprises, consists of, or consists essentially of a CDR-L2 sequence of an illustrative antibody or V_L sequence provided herein. In some aspects, the CDR-L2 sequence is a CDR-L2 sequence of a V_L sequence provided in SEQ ID NOs.: 367-369.

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 302-304. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 302. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 303. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L2 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 304.

2.5.3. CDR-L1

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L1 sequence, wherein the CDR-L1 sequence comprises, consists of, or consists essentially of a CDR-L1 sequence of an illustrative antibody or V_L sequence provided herein. In some aspects, the CDR-L1 sequence is a CDR-L1 sequence of a V_L sequence provided in SEQ ID NOs.: 367-369.

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 299-301. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 299. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 300. In some aspects, the antibody comprises a V_L sequence comprising a CDR-L1 sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 301.

2.5.4. CDR-L3+CDR-L2

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 305-307 and a CDR-L2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 302-304. In some aspects, the CDR-L3 sequence and the CDR-L2 sequence are both from a single illustrative V_L sequence provided in this disclosure. For example, in some aspects, the CDR-L3 and CDR-L2 are both from a single illustrative V_L sequence selected from SEQ ID NOs.: 367-369.

2.5.5. CDR-L3+CDR-L1

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 305-307 and a CDR-L1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 299-301. In some aspects, the CDR-L3 sequence and the CDR-L1 sequence are both from a single illustrative V_L sequence provided in this disclosure. For example, in some aspects, the CDR-L3 and CDR-L1 are both from a single illustrative V_L sequence selected from SEQ ID NOs.: 367-369.

2.5.6. CDR-L1+CDR-L2

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 299-301 and a CDR-L2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 302-304. In some aspects, the CDR-L1 sequence and the CDR-L2 sequence are both from a single illustrative V_L sequence provided in this disclosure. For example, in some aspects, the CDR-L1 and CDR-L2 are both from a single illustrative V_L sequence selected from SEQ ID NOs.: 367-369.

2.5.7. CDR-L1+CDR-L2+CDR-L3

In some embodiments, the antibody comprises a V_L sequence comprising a CDR-L1 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 299-301, a CDR-L2 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 302-304, and a CDR-L3 sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs: 305-307. In some aspects, the CDR-L1 sequence, CDR-L2 sequence, and CDR-L3 sequence are all from a single illustrative V_L sequence provided in this disclosure. For example, in some aspects, the CDR-L1, CDR-L2, and CDR-L3 are all from a single illustrative V_L sequence selected from SEQ ID NOs.: 367-369.

2.5.8. Variants of V_L Sequences Comprising Illustrative CDR-Ls

In some embodiments, the V_L sequences provided herein comprise a variant of an illustrative CDR-L3, CDR-L2, and/or CDR-L1 sequence provided in this disclosure.

In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L3 sequence provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L3 sequences provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L2 sequence provided in this disclosure. In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L2 sequences provided in this disclosure. In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L2 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L1 sequence provided in this disclosure. In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L1 sequences provided in this disclosure. In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L1 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.6. V_L Sequences

In some embodiments, the antibody comprises, consists of, or consists essentially of a V_L sequence provided in SEQ ID NOs.: 367-369.

In some embodiments, the antibody comprises a V_L sequence comprising, consisting of, or consisting essentially of a sequence selected from SEQ ID NOs.: 367-369. In some aspects, the antibody comprises a V_L sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 367. In some aspects, the antibody comprises a V_L sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 368. In some aspects, the antibody comprises a V_L sequence comprising, consisting of, or consisting essentially of SEQ ID NO: 369.

2.6.1. Variants of V_L Sequences

In some embodiments, the V_L sequences provided herein comprise, consist of, or consist essentially of a variant of an illustrative V_L sequence provided in this disclosure.

In some aspects, the V_L sequence comprises, consists of, or consists essentially of a variant of an illustrative V_L sequence provided in this disclosure. In some aspects, the V_L sequence comprises, consists of, or consists essentially of a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with any of the illustrative V_L sequences provided in this disclosure.

In some embodiments, the V_L sequence comprises, consists of, or consists essentially of any of the illustrative V_L sequences provided in this disclosure having 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer, 16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 or fewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.7. Pairs

2.7.1. CDR-H3-CDR-L3 Pairs

In some embodiments, the antibody comprises a CDR-H3 sequence and a CDR-L3 sequence. In some aspects, the CDR-H3 sequence is part of a V_H and the CDR-L3 sequence is part of a V_L.

In some aspects, the CDR-H3 sequence is a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 240-298, and the CDR-L3 sequence is a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 305-307.

In some aspects, the CDR-H3-CDR-L3 pairs are selected from SEQ ID NO: 305 and SEQ ID NO: 240; SEQ ID NO: 305 and SEQ ID NO: 241; SEQ ID NO: 305 and SEQ ID NO: 242; SEQ ID NO: 305 and SEQ ID NO: 243; SEQ ID NO: 305 and SEQ ID NO: 244; SEQ ID NO: 305 and SEQ ID NO: 245; SEQ ID NO: 305 and SEQ ID NO: 246; SEQ ID NO: 305 and SEQ ID NO: 247; SEQ ID NO: 305 and SEQ ID NO: 248; SEQ ID NO: 305 and SEQ ID NO: 249; SEQ ID NO: 305 and SEQ ID NO: 250; SEQ ID NO: 305 and SEQ ID NO: 251; SEQ ID NO: 305 and SEQ ID NO: 252; SEQ ID NO: 305 and SEQ ID NO: 253; SEQ ID NO: 305 and SEQ ID NO: 254; SEQ ID NO: 305 and SEQ ID NO: 255; SEQ ID NO: 305 and SEQ ID NO: 256; SEQ ID NO: 305 and SEQ ID NO: 257; SEQ ID NO: 305 and SEQ ID NO: 258; SEQ ID NO: 305 and SEQ ID NO: 259; SEQ ID NO: 305 and SEQ ID NO: 260; SEQ ID NO: 305 and SEQ ID NO: 261; SEQ ID NO: 305 and SEQ ID NO: 262; SEQ ID NO: 305 and SEQ ID NO: 263; SEQ ID NO: 305 and SEQ ID NO: 264; SEQ ID NO: 305 and SEQ ID NO: 265; SEQ ID NO: 305 and SEQ ID NO: 266; SEQ ID NO: 305 and SEQ ID NO: 267; SEQ ID NO: 305 and SEQ ID NO: 268; SEQ ID NO: 305 and SEQ ID NO: 269; SEQ ID NO: 305 and SEQ ID NO: 270; SEQ ID NO: 305 and SEQ ID NO: 271; SEQ ID NO: 305 and SEQ ID NO: 272; SEQ ID NO: 305 and SEQ ID NO: 273; SEQ ID NO: 305 and SEQ ID NO: 274; SEQ ID NO: 305 and SEQ ID NO: 275; SEQ ID NO: 305 and SEQ ID NO: 276; SEQ ID NO: 305 and SEQ ID NO: 277; SEQ ID NO: 305 and SEQ ID NO: 278; SEQ ID NO: 305 and SEQ ID NO: 279; SEQ ID NO: 305 and SEQ ID NO: 280; SEQ ID NO: 305 and SEQ ID NO: 281; SEQ ID NO: 305 and SEQ ID NO: 282; SEQ ID NO: 305 and SEQ ID NO: 283; SEQ ID NO: 305 and SEQ ID NO: 284; SEQ ID NO: 305 and SEQ ID NO: 285; SEQ ID NO: 305 and SEQ ID NO: 286; SEQ ID NO: 305 and SEQ ID NO: 287; SEQ ID NO: 305 and SEQ ID NO: 288; SEQ ID NO: 305 and SEQ ID NO: 289; SEQ ID NO: 305 and SEQ ID NO: 290; SEQ ID NO: 305 and SEQ ID NO: 291; SEQ ID NO: 305 and SEQ ID NO: 292; SEQ ID NO: 305 and SEQ ID NO: 293; SEQ ID NO: 305 and SEQ ID NO: 294; SEQ ID NO: 305 and SEQ ID NO: 295; SEQ ID NO: 305 and SEQ ID NO: 296; SEQ ID NO: 305 and SEQ ID NO: 297; and SEQ ID NO: 305 and SEQ ID NO: 298.

In some aspects, the CDR-H3-CDR-L3 pairs are selected from SEQ ID NO: 306 and SEQ ID NO: 240; SEQ ID NO: 306 and SEQ ID NO: 241; SEQ ID NO: 306 and SEQ ID NO: 242; SEQ ID NO: 306 and SEQ ID NO: 243; SEQ ID NO: 306 and SEQ ID NO: 244; SEQ ID NO: 306 and SEQ ID NO: 245; SEQ ID NO: 306 and SEQ ID NO: 246; SEQ ID NO: 306 and SEQ ID NO: 247; SEQ ID NO: 306 and SEQ ID NO: 248; SEQ ID NO: 306 and SEQ ID NO: 249; SEQ ID NO: 306 and SEQ ID NO: 250; SEQ ID NO: 306 and SEQ ID NO: 251; SEQ ID NO: 306 and SEQ ID NO: 252; SEQ ID NO: 306 and SEQ ID NO: 253; SEQ ID NO: 306 and SEQ ID NO: 254; SEQ ID NO: 306 and SEQ ID NO: 255; SEQ ID NO: 306 and SEQ ID NO: 256; SEQ ID NO: 306 and SEQ ID NO: 257; SEQ ID NO: 306 and SEQ ID NO: 258; SEQ ID NO: 306 and SEQ ID NO: 259; SEQ ID NO: 306 and SEQ ID NO: 260; SEQ ID NO: 306 and SEQ ID NO: 261; SEQ ID NO: 306 and SEQ ID NO: 262; SEQ ID NO: 306 and SEQ ID NO: 263; SEQ ID NO: 306 and SEQ ID NO: 264; SEQ ID NO: 306 and SEQ ID NO: 265; SEQ ID NO: 306 and SEQ ID NO: 266; SEQ ID NO: 306 and SEQ ID NO: 267; SEQ ID NO: 306 and SEQ ID NO: 268; SEQ ID NO: 306 and SEQ ID NO: 269; SEQ ID NO: 306 and SEQ ID NO: 270; SEQ ID NO: 306 and SEQ ID NO: 271; SEQ ID NO: 306 and SEQ ID NO: 272; SEQ ID NO: 306 and SEQ ID NO: 273; SEQ ID NO: 306 and SEQ ID NO: 274; SEQ ID NO: 306 and SEQ ID NO: 275; SEQ ID NO: 306 and SEQ ID NO: 276; SEQ ID NO: 306 and SEQ ID NO: 277; SEQ ID NO: 306 and SEQ ID NO: 278; SEQ ID NO: 306 and SEQ ID NO: 279; SEQ ID NO: 306 and SEQ ID NO: 280; SEQ ID NO: 306 and SEQ ID NO: 281; SEQ ID NO: 306 and SEQ ID NO: 282; SEQ ID NO: 306 and SEQ ID NO: 283; SEQ ID NO: 306 and SEQ ID NO: 284; SEQ ID NO: 306 and SEQ ID NO: 285; SEQ ID NO: 306 and SEQ ID NO: 286; SEQ ID NO: 306 and SEQ ID NO: 287; SEQ ID NO: 306 and SEQ ID NO: 288; SEQ ID NO: 306 and SEQ ID NO: 289; SEQ ID NO: 306 and SEQ ID NO: 290; SEQ ID NO: 306 and SEQ ID NO: 291; SEQ ID NO: 306 and SEQ ID NO: 292; SEQ ID NO: 306 and SEQ ID NO: 293; SEQ ID NO: 306 and SEQ ID NO: 294; SEQ ID NO: 306 and SEQ ID NO: 295; SEQ ID NO: 306 and SEQ ID NO: 296; SEQ ID NO: 306 and SEQ ID NO: 297; and SEQ ID NO: 306 and SEQ ID NO: 298.

In some aspects, the CDR-H3-CDR-L3 pairs are selected from SEQ ID NO: 307 and SEQ ID NO: 240; SEQ ID NO: 307 and SEQ ID NO: 241; SEQ ID NO: 307 and SEQ ID NO: 242; SEQ ID NO: 307 and SEQ ID NO: 243; SEQ ID NO: 307 and SEQ ID NO: 244; SEQ ID NO: 307 and SEQ ID NO: 245; SEQ ID NO: 307 and SEQ ID NO: 246; SEQ ID NO: 307 and SEQ ID NO: 247; SEQ ID NO: 307 and SEQ ID NO: 248; SEQ ID NO: 307 and SEQ ID NO: 249; SEQ ID NO: 307 and SEQ ID NO: 250; SEQ ID NO: 307 and SEQ ID NO: 251; SEQ ID NO: 307 and SEQ ID NO: 252; SEQ ID NO: 307 and SEQ ID NO: 253; SEQ ID NO: 307 and SEQ ID NO: 254; SEQ ID NO: 307 and SEQ ID NO: 255; SEQ ID NO: 307 and SEQ ID NO: 256; SEQ ID NO: 307 and SEQ ID NO: 257; SEQ ID NO: 307 and SEQ ID NO: 258; SEQ ID NO: 307 and SEQ ID NO: 259; SEQ ID NO: 307 and SEQ ID NO: 260; SEQ ID NO: 307 and SEQ ID NO: 261; SEQ ID NO: 307 and SEQ ID NO: 262; SEQ ID NO: 307 and SEQ ID NO: 263; SEQ ID NO: 307 and SEQ ID NO: 264; SEQ ID NO: 307 and SEQ ID NO: 265; SEQ ID NO: 307 and SEQ ID NO: 266; SEQ ID NO: 307 and SEQ ID NO: 267; SEQ ID NO: 307 and SEQ ID NO: 268; SEQ ID NO: 307 and SEQ ID NO: 269; SEQ ID NO: 307 and SEQ ID NO: 270; SEQ ID NO: 307 and SEQ ID NO: 271; SEQ ID NO: 307 and SEQ ID NO: 272; SEQ ID NO: 307 and SEQ ID NO: 273; SEQ ID NO: 307 and SEQ ID NO: 274; SEQ ID NO: 307 and SEQ ID NO: 275; SEQ ID NO: 307 and SEQ ID NO: 276; SEQ ID NO: 307 and SEQ ID NO: 277; SEQ ID NO: 307 and SEQ ID NO: 278; SEQ ID NO: 307 and SEQ ID NO: 279; SEQ ID NO: 307 and SEQ ID NO: 280; SEQ ID NO: 307 and SEQ ID NO: 281; SEQ ID NO: 307 and SEQ ID NO: 282; SEQ ID NO: 307 and SEQ ID NO: 283; SEQ ID NO: 307 and SEQ ID NO: 284; SEQ ID NO: 307 and SEQ ID NO: 285; SEQ ID NO: 307 and SEQ ID NO: 286; SEQ ID NO: 307 and SEQ ID NO: 287; SEQ ID NO: 307 and SEQ ID NO: 288; SEQ ID NO: 307 and SEQ ID NO: 289; SEQ ID NO: 307 and SEQ ID NO: 290; SEQ ID NO: 307 and SEQ ID NO: 291; SEQ ID NO: 307 and SEQ ID NO: 292; SEQ ID NO: 307 and SEQ ID NO: 293; SEQ ID NO: 307 and SEQ ID NO: 294; SEQ ID NO: 307 and SEQ ID NO: 295; SEQ ID NO: 307 and SEQ ID NO: 296; SEQ ID NO: 307 and SEQ ID NO: 297; and SEQ ID NO: 307 and SEQ ID NO: 298.

2.7.1.1. Variants of CDR-H3-CDR-L3 Pairs

In some embodiments, the CDR-H3-CDR-L3 pairs provided herein comprise a variant of an illustrative CDR-H3 and/or CDR-L1 sequence provided in this disclosure.

In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-H3 sequence provided in this disclosure. In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-H3 sequences provided in this disclosure. In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-H3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L3 sequence provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L3 sequences provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.7.2. CDR-H1-CDR-L1 Pairs

In some embodiments, the antibody comprises a CDR-H1 sequence and a CDR-L1 sequence. In some aspects, the CDR-H1 sequence is part of a V_H and the CDR-L1 sequence is part of a V_L.

In some aspects, the CDR-H1 sequence is a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 4-62, and the CDR-L1 sequence is a CDR-L1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 299-301.

In some aspects, the CDR-H1 sequence is a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 63-121, and the CDR-L1 sequence is a CDR-L1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 299-301.

2.7.2.1. Variants of CDR-H1-CDR-L1 Pairs

In some embodiments, the CDR-H1-CDR-L1 pairs provided herein comprise a variant of an illustrative CDR-H1 and/or CDR-L1 sequence provided in this disclosure.

In some aspects, the CDR-H1 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-H1 sequence provided in this disclosure. In some aspects, the CDR-H1 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-H1 sequences provided in this disclosure. In some aspects, the CDR-H1 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-H1 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L1 sequence provided in this disclosure. In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L1 sequences provided in this disclosure. In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L1 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.7.3. CDR-H2-CDR-L2 Pairs

In some embodiments, the antibody comprises a CDR-H2 sequence and a CDR-L2 sequence. In some aspects, the CDR-H2 sequence is part of a V_H and the CDR-L2 sequence is part of a V_L.

In some aspects, the CDR-H2 sequence is a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 122-180, and the CDR-L2 sequence is a CDR-L2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 302-304.

In some aspects, the CDR-H1 sequence is a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 181-239, and the CDR-L2 sequence is a CDR-L2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 302-304.

2.7.3.1. Variants of CDR-H2-CDR-L2 Pairs

In some embodiments, the CDR-H2-CDR-L2 pairs provided herein comprise a variant of an illustrative CDR-H2 and/or CDR-L2 sequence provided in this disclosure.

In some aspects, the CDR-H2 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-H2 sequence provided in this disclosure. In some aspects, the CDR-H2 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-H2 sequences provided in this disclosure. In some aspects, the CDR-H2 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-H2 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L2 sequence provided in this disclosure. In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L2 sequences provided in this disclosure. In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L2 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.7.4. V_H-V_L Pairs

In some embodiments, the antibody comprises a V_H sequence and a V_L sequence.

In some aspects, the V_H sequence is a V_H sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 308-366, and the V_L sequence is a V_L sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 367-369.

In some aspects, the V_H-V_L pairs are selected from SEQ ID NO: 367 and SEQ ID NO: 308; SEQ ID NO: 367 and SEQ ID NO: 309; SEQ ID NO: 367 and SEQ ID NO: 310; SEQ ID NO: 367 and SEQ ID NO: 311; SEQ ID NO: 367 and SEQ ID NO: 312; SEQ ID NO: 367 and SEQ ID NO: 313; SEQ ID NO: 367 and SEQ ID NO: 314; SEQ ID NO: 367 and SEQ ID NO: 315; SEQ ID NO: 367 and SEQ ID NO: 316; SEQ ID NO: 367 and SEQ ID NO: 317; SEQ ID NO: 367 and SEQ ID NO: 318; SEQ ID NO: 367 and SEQ ID NO: 319; SEQ ID NO: 367 and SEQ ID NO: 320; SEQ ID NO: 367 and SEQ ID NO: 321; SEQ ID NO: 367 and SEQ ID NO: 322; SEQ ID NO: 367 and SEQ ID NO: 323; SEQ ID NO: 367 and SEQ ID NO: 324; SEQ ID NO: 367 and SEQ ID NO: 325; SEQ ID NO: 367 and SEQ ID NO: 326; SEQ ID NO: 367 and SEQ ID NO: 327; SEQ ID NO: 367 and SEQ ID NO: 328; SEQ ID NO: 367 and SEQ ID NO: 329; SEQ ID NO: 367 and SEQ ID NO: 330; SEQ ID NO: 367 and SEQ ID NO: 331; SEQ ID NO: 367 and SEQ ID NO: 332; SEQ ID NO: 367 and SEQ ID NO: 333; SEQ ID NO: 367 and SEQ ID NO: 334; SEQ ID NO: 367 and SEQ ID NO: 335; SEQ ID NO: 367 and SEQ ID NO: 336; SEQ ID NO: 367 and SEQ ID NO: 337; SEQ ID NO: 367 and SEQ ID NO: 338; SEQ ID NO: 367 and SEQ ID NO: 339; SEQ ID NO: 367 and SEQ ID NO: 340; SEQ ID NO: 367 and SEQ ID NO: 341; SEQ ID NO: 367 and SEQ ID NO: 342; SEQ ID NO: 367 and SEQ ID NO: 343; SEQ ID NO: 367 and SEQ ID NO: 344; SEQ ID NO: 367 and SEQ ID NO: 345; SEQ ID NO: 367 and SEQ ID NO: 346; SEQ ID NO: 367 and SEQ ID NO: 347; SEQ ID NO: 367 and SEQ ID NO: 348; SEQ ID NO: 367 and SEQ ID NO: 349; SEQ ID NO: 367 and SEQ ID NO: 350; SEQ ID NO: 367 and SEQ ID NO: 351; SEQ ID NO: 367 and SEQ ID NO: 352; SEQ ID NO: 367 and SEQ ID NO: 353; SEQ ID NO: 367 and SEQ ID NO: 354; SEQ ID NO: 367 and SEQ ID NO: 355; SEQ ID NO: 367 and SEQ ID NO: 356; SEQ ID NO: 367 and SEQ ID NO: 357; SEQ ID NO: 367 and SEQ ID NO: 358; SEQ ID NO: 367 and SEQ ID NO: 359; SEQ ID NO: 367 and SEQ ID NO: 360; SEQ ID NO: 367 and SEQ ID NO: 361; SEQ ID NO: 367 and SEQ ID NO: 362; SEQ ID NO: 367 and SEQ ID NO: 363; SEQ ID NO: 367 and SEQ ID NO: 364; SEQ ID NO: 367 and SEQ ID NO: 365; and SEQ ID NO: 367 and SEQ ID NO: 366.

In some aspects, the V_H-V_L pairs are selected from SEQ ID NO: 368 and SEQ ID NO: 308; SEQ ID NO: 368 and SEQ ID NO: 309; SEQ ID NO: 368 and SEQ ID NO: 310; SEQ ID NO: 368 and SEQ ID NO: 311; SEQ ID NO: 368 and SEQ ID NO: 312; SEQ ID NO: 368 and SEQ ID NO: 313; SEQ ID NO: 368 and SEQ ID NO: 314; SEQ ID NO: 368 and SEQ ID NO: 315; SEQ ID NO: 368 and SEQ ID NO: 316; SEQ ID NO: 368 and SEQ ID NO: 317; SEQ ID NO: 368 and SEQ ID NO: 318; SEQ ID NO: 368 and SEQ ID NO: 319; SEQ ID NO: 368 and SEQ ID NO: 320; SEQ ID NO: 368 and SEQ ID NO: 321; SEQ ID NO: 368 and SEQ ID NO: 322; SEQ ID NO: 368 and SEQ ID NO: 323; SEQ ID NO: 368 and SEQ ID NO: 324; SEQ ID NO: 368 and SEQ ID NO: 325; SEQ ID NO: 368 and SEQ ID NO: 326; SEQ ID NO: 368 and SEQ ID NO: 327; SEQ ID NO: 368 and SEQ ID NO: 328; SEQ ID NO: 368 and SEQ ID NO: 329; SEQ ID NO: 368 and SEQ ID NO: 330; SEQ ID NO: 368 and SEQ ID NO: 331; SEQ ID NO: 368 and SEQ ID NO: 332; SEQ ID NO: 368 and SEQ ID NO: 333; SEQ ID NO: 368 and SEQ ID NO: 334; SEQ ID NO: 368 and SEQ ID NO: 335; SEQ ID NO: 368 and SEQ ID NO: 336; SEQ ID NO: 368 and SEQ ID NO: 337; SEQ ID NO: 368 and SEQ ID NO: 338; SEQ ID NO: 368 and SEQ ID NO: 339; SEQ ID NO: 368 and SEQ ID NO: 340; SEQ ID NO: 368 and SEQ ID NO: 341; SEQ ID NO: 368 and SEQ ID NO: 342; SEQ ID NO: 368 and SEQ ID NO: 343; SEQ ID NO: 368 and SEQ ID NO: 344; SEQ ID NO: 368 and SEQ ID NO: 345; SEQ ID NO: 368 and SEQ ID NO: 346; SEQ ID NO: 368 and SEQ ID NO: 347; SEQ ID NO: 368 and SEQ ID NO: 348; SEQ ID NO: 368 and SEQ ID NO: 349; SEQ ID NO: 368 and SEQ ID NO: 350; SEQ ID NO: 368 and SEQ ID NO: 351; SEQ ID NO: 368 and SEQ ID NO: 352; SEQ ID NO: 368 and SEQ ID NO: 353; SEQ ID NO: 368 and SEQ ID NO: 354; SEQ ID NO: 368 and SEQ ID NO: 355; SEQ ID NO: 368 and SEQ ID NO: 356; SEQ ID NO: 368 and SEQ ID NO: 357; SEQ ID NO: 368 and SEQ ID NO: 358; SEQ ID NO: 368 and SEQ ID NO: 359; SEQ ID NO: 368 and SEQ ID NO: 360; SEQ ID NO: 368 and SEQ ID NO: 361; SEQ ID NO: 368 and SEQ ID NO: 362; SEQ ID NO: 368 and SEQ ID NO: 363; SEQ ID NO: 368 and SEQ ID NO: 364; SEQ ID NO: 368 and SEQ ID NO: 365; and SEQ ID NO: 368 and SEQ ID NO: 366.

In some aspects, the V_H-V_L pairs are selected from SEQ ID NO: 369 and SEQ ID NO: 308; SEQ ID NO: 369 and SEQ ID NO: 309; SEQ ID NO: 369 and SEQ ID NO: 310; SEQ ID NO: 369 and SEQ ID NO: 311; SEQ ID NO: 369 and SEQ ID NO: 312; SEQ ID NO: 369 and SEQ ID NO: 313; SEQ ID NO: 369 and SEQ ID NO: 314; SEQ ID NO: 369 and SEQ ID NO: 315; SEQ ID NO: 369 and SEQ ID NO: 316; SEQ ID NO: 369 and SEQ ID NO: 317; SEQ ID NO: 369 and SEQ ID NO: 318; SEQ ID NO: 369 and SEQ ID NO: 319; SEQ ID NO: 369 and SEQ ID NO: 320; SEQ ID NO: 369 and SEQ ID NO: 321; SEQ ID NO: 369 and SEQ ID NO: 322; SEQ ID NO: 369 and SEQ ID NO: 323; SEQ ID NO: 369 and SEQ ID NO: 324; SEQ ID NO: 369 and SEQ ID NO: 325; SEQ ID NO: 369 and SEQ ID NO: 326; SEQ ID NO: 369 and SEQ ID NO: 327; SEQ ID NO: 369 and SEQ ID NO: 328; SEQ ID NO: 369 and SEQ ID NO: 329; SEQ ID NO: 369 and SEQ ID NO: 330; SEQ ID NO: 369 and SEQ ID NO: 331; SEQ ID NO: 369 and SEQ ID NO: 332; SEQ ID NO: 369 and SEQ ID NO: 333; SEQ ID NO: 369 and SEQ ID NO: 334; SEQ ID NO: 369 and SEQ ID NO: 335; SEQ ID NO: 369 and SEQ ID NO: 336; SEQ ID NO: 369 and SEQ ID NO: 337; SEQ ID NO: 369 and SEQ ID NO: 338; SEQ ID NO: 369 and SEQ ID NO: 339; SEQ ID NO: 369 and SEQ ID NO: 340; SEQ ID NO: 369 and SEQ ID NO: 341; SEQ ID NO: 369 and SEQ ID NO: 342; SEQ ID NO: 369 and SEQ ID NO: 343; SEQ ID NO: 369 and SEQ ID NO: 344; SEQ ID NO: 369 and SEQ ID NO: 345; SEQ ID NO: 369 and SEQ ID NO: 346; SEQ ID NO: 369 and SEQ ID NO: 347; SEQ ID NO: 369 and SEQ ID NO: 348; SEQ ID NO: 369 and SEQ ID NO: 349; SEQ ID NO: 369 and SEQ ID NO: 350; SEQ ID NO: 369 and SEQ ID NO: 351; SEQ ID NO: 369 and SEQ ID NO: 352; SEQ ID NO: 369 and SEQ ID NO: 353; SEQ ID NO: 369 and SEQ ID NO: 354; SEQ ID NO: 369 and SEQ ID NO: 355; SEQ ID NO: 369 and SEQ ID NO: 356; SEQ ID NO: 369 and SEQ ID NO: 357; SEQ ID NO: 369 and SEQ ID NO: 358; SEQ ID NO: 369 and SEQ ID NO: 359; SEQ ID NO: 369 and SEQ ID NO: 360; SEQ ID NO: 369 and SEQ ID NO: 361; SEQ ID NO: 369 and SEQ ID NO: 362; SEQ ID NO: 369 and SEQ ID NO: 363; SEQ ID NO: 369 and SEQ ID NO: 364; SEQ ID NO: 369 and SEQ ID NO: 365; and SEQ ID NO: 369 and SEQ ID NO: 366.

2.7.4.1. Variants of V_H-V_L Pairs

In some embodiments, the V_H-V_L pairs provided herein comprise a variant of an illustrative V_H and/or V_L sequence provided in this disclosure.

In some aspects, the Vii sequence comprises, consists of, or consists essentially of a variant of an illustrative V_H sequence provided in this disclosure. In some aspects, the V_H sequence comprises, consists of, or consists essentially of a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.1% identity with any of the illustrative Vii sequences provided in this disclosure.

In some embodiments, the V_H sequence comprises, consists of, or consists essentially of any of the illustrative Vii sequences provided in this disclosure having 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer, 16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 or fewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the V_L sequence comprises, consists of, or consists essentially of a variant of an illustrative V_L sequence provided in this disclosure. In some aspects, the V_L sequence comprises, consists of, or consists essentially of a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with any of the illustrative V_L sequences provided in this disclosure.

In some embodiments, the V_L sequence comprises, consists of, or consists essentially of any of the illustrative V_L sequences provided in this disclosure having 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer, 16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 or fewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.8. Antibodies Comprising All Six CDRs

In some embodiments, the antibody comprises a CDR-H1 sequence, a CDR-H2 sequence, a CDR-H3 sequence, a CDR-L1 sequence, and a CDR-L3 sequence. In some aspects, the CDR sequences are part of a V_H (for CDR-H) or V_L (for CDR-L).

In some aspects, the CDR-H1 sequence is a Chothia CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 4-62; the CDR-H2 sequence is a Chothia CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 122-180; the CDR-H3 sequence is a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 240-298; the CDR-L1 sequence is a CDR-L1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 299-301; the CDR-L2 sequence is a CDR-L2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 302-304; and the CDR-L3 sequence is a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 305-307.

In some aspects, the CDR-H1 sequence is a Kabat CDR-H1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 63-121; the CDR-H2 sequence is a Kabat CDR-H2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 181-239; the CDR-H3 sequence is a CDR-H3 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 240-298; the CDR-L1 sequence is a CDR-L1 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 299-301; the CDR-L2 sequence is a CDR-L2 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 302-304; and the CDR-L3 sequence is a CDR-L3 sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 305-307.

2.8.1. Variants of Antibodies Comprising All Six CDRs

In some embodiments, the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 provided herein comprise a variant of an illustrative CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and/or CDR-L3 sequence provided in this disclosure.

In some aspects, the CDR-H1 sequence comprises, consists of, or consists essentially of a variant of an illustrative Chothia or Kabat CDR-H1 sequence provided in this disclosure. In some aspects, the CDR-H1 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Chothia or Kabat CDR-H1 sequences provided in this disclosure. In some aspects, the CDR-H1 sequence comprises, consists of, or consists essentially of any of the illustrative Chothia or Kabat CDR-H1 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-H2 sequence comprises, consists of, or consists essentially of a variant of an illustrative Chothia or Kabat CDR-H2 sequence provided in this disclosure. In some aspects, the CDR-H2 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative Chothia or Kabat CDR-H2 sequences provided in this disclosure. In some aspects, the CDR-H2 sequence comprises, consists of, or consists essentially of any of the illustrative Chothia or Kabat CDR-H2 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-H3 sequence provided in this disclosure. In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-H3 sequences provided in this disclosure. In some aspects, the CDR-H3 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-H3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L1 sequence provided in this disclosure. In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L1 sequences provided in this disclosure. In some aspects, the CDR-L1 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L1 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L2 sequence provided in this disclosure. In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L2 sequences provided in this disclosure. In some aspects, the CDR-L2 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L2 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a variant of an illustrative CDR-L3 sequence provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of a sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L3 sequences provided in this disclosure. In some aspects, the CDR-L3 sequence comprises, consists of, or consists essentially of any of the illustrative CDR-L3 sequences provided in this disclosure, with 1, 2, or 3 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions.

2.9. Consensus Sequences

In some embodiments, provided herein are anti-FOLR1 antibodies comprising one or more sequences defined by consensus sequences. Each consensus sequence is based, at least in part, on one or more alignments of two or more useful anti-FOLR1 CDR sequences provided in this disclosure. Based on such alignments, a person of skill in the art would recognize that different amino acid residues may useful in certain positions of the CDRs. Accordingly, each consensus sequence encompasses two or more useful anti-FOLR1 CDR sequences.

In some embodiments, the antibodies comprise one to six of the consensus CDR sequences provided herein. In some embodiments, the antibodies comprise two to six of the consensus CDR sequences provided herein. In some embodiments, the antibodies comprise three to six of the consensus CDR sequences provided herein. In some embodiments, the antibodies comprise four to six of the consensus CDR sequences provided herein. In some embodiments, the antibodies comprise five to six of the consensus CDR sequences provided herein. In some embodiments, the antibodies comprise six of the consensus CDR sequences provided herein. In some embodiments, the antibodies comprise a V_L comprising the CDR-L consensus sequence(s). In some embodiments, the antibodies comprise a V_H comprising the CDR-H consensus sequence(s). In some embodiments, the antibodies comprise a V_H comprising the CDR-H consensus sequence(s) and a V_L comprising the CDR-L consensus sequence(s).

2.9.1. CDR-H3 Consensus Sequences

In some embodiments, the antibody comprises a CDR-H3 sequence defined by the consensus sequence G-α₂-α₃-α₄-W-α₆-α₇-G-α₉-α₁₀-Y-α₁₂-α₁₃-α₁₄-Y, where α₂ is G, S, A, F, H, R, T, or Y; α₃ is W, L, or Y; α₄ is S, A, F, Y, H, or D; as is R, P, Q, or K; α₇ is S, A, or H; α₉ is Y, H, or M; α₁₀ is G, S, D, or W; α₁₂ is Y or F; α₁₃ is L, I, Q, or M; and α₁₄ is D or E.

In some embodiments, the antibody comprises a CDR-H3 sequence defined by the consensus sequence G-α₂-α₃-α₄-W-α₆-α₇-G-α₉-α₁₀-Y-α₁₂-α₁₃-α₁₄-Y, where α₂ is G or S; α₃ is W; α₄ is S or H; as is R or P; α₇ is S; α₉ is Y or M; α₁₀ is G, S, or D; α₁₂ is Y; α₁₃ is L; and α₁₄ is D.

2.9.2. Chothia CDR-H1 Consensus Sequences

In some embodiments, the antibody comprises a Chothia CDR-H1 sequence defined by the consensus sequence γ₁-γ₂-γ₃-γ₄-γ₅-γ₆-γ₇, where γ₁ is G or S; γ₂ is F or S; γ₃ is N; γ₄ is I or T; γ₅ is S, R, G, T, N, or D; γ₆ is N, K, T, R, H, Y, L, M, Q, or V; and γ₇ is Y, H, S, N, K, F, or Q.

In some embodiments, the antibody comprises a Chothia CDR-H1 sequence defined by the consensus sequence γ₁-γ₂-γ₃-γ₄-γ₅-γ₆-γ₇, where γ₁ is G; γ₂ is F; γ₃ is N; γ₄ is I or T; γ₅ is S, R, or T; γ₆ is N or T; and γ₇ is Y, K, or Q.

2.9.3. Chothia CDR-H2 Consensus Sequences

In some embodiments, the antibody comprises a Chothia CDR-H2 sequence defined by the consensus sequence ε₁-ε₂-ε₃-ε₄-ε₅-ε₆, where ε₁ is Y, T, F, S, or A; ε₂ is P; ε₃ is N, I, V, R, Y, F, G, L, Q, or S; ε₄ is D or P; ε₅ is G or D; and ε₆ is Y, I, T, N, F, S, or M.

In some embodiments, the antibody comprises a Chothia CDR-H2 sequence defined by the consensus sequence ε₁-ε₂-ε₃-ε₄-ε₅-ε₆, where ε₁ is Y or F; ε₂ is P; ε₃ is N, I, or R; ε₄ is D; ε₅ is G; and ε₆ is Y or I.

2.9.4. Kabat CDR-H1 Consensus Sequences

In some embodiments, the antibody comprises a Kabat CDR-H1 sequence defined by the consensus sequence ζ₁-ζ₂-ζ₃-ζ₄-ζ₅, where ζ₁ is N, K, T, R, H, Y, L, M, Q, or V; ζ₂ is Y, H, S, N, K, F, or Q; ζ₃ is S or Y; ζ₄ is I; and ζ₅ is H.

In some embodiments, the antibody comprises a Kabat CDR-H1 sequence defined by the consensus sequence ζ₁-ζ₂-ζ₃-ζ₄-ζ₅, where ζ₁ is N or T; ζ₂ is Y, K, or Q; ζ₃ is S; ζ₄ is I; and ζ₅ is H.

2.9.5. Kabat CDR-H2 Consensus Sequences

In some embodiments, the antibody comprises a Kabat CDR-H2 sequence defined by the consensus sequence θ₁-θ₂-θ₃-θ₄-θ₅-θ₆-θ₇-θ₈-θ₉-D-Y-A-D-θ₁₄-θ₁₅-θ₁₆-G, where θ₁ is G, E, D, W, S, or V; θ₂ is I or V; θ₃ is Y, T, F, S, or A; θ₄ is P; θ₅ is N, I, V, R, Y, F, G, L, Q, or S; θ₆ is D or P; θ₇ is G or D; θ₈ is Y, I, T, N, F, S, or M; θ₉ is T or N; θ₁₄ is S, R, or N; θ₁₅ is V or M; and θ₁₆ is K or E.

In some embodiments, the antibody comprises a Kabat CDR-H2 sequence defined by the consensus sequence θ₁-θ₂-θ₃-θ₄-θ₅-θ₆-θ₇-θ₈-θ₉-D-Y-A-D-θ₁₄-θ₁₅-θ₁₆-G, where θ₁ is G, E, or D; θ₂ is I; θ₃ is Y or F; θ₄ is P; θ₅ is N, I, or R; θ₆ is D; θ₇ is G; θ₈ is Y or I; θ₉ is T; θ₁₄ is S; θ₁₅ is V; and θ₁₆ is K.

3. Germline

In some embodiments, the antibody that specifically binds folate receptor alpha is an antibody comprising a variable region that is encoded by a particular germline gene, or a variant thereof. The illustrative antibodies provided herein comprise variable regions that are encoded by the heavy chain variable region germline genes VH1-18, VH3-33 VH2-5, V_H2-70, and VH4-30-4. or variants thereof; and the light chain variable region germline genes Vκ1-5, Vκ3-11, Vκ2-20, Vκ1-33, and Vκ1-16, or variants thereof.

One of skill in the art would recognize that the CDR sequences provided herein may also be useful when combined with variable regions encoded by other variable region germline genes, or variants thereof. In particular, the CDR sequences provided herein may be useful when combined with variable regions encoded by variable region germline genes, or variants thereof, that are structurally similar to the variable region germline genes recited above. For example, in some embodiments, a CDR-H sequence provided herein may be combined with a variable region encoded by a variable region germline gene selected from the V_H1, V_H2, V_H3, or V_H4 families, or a variant thereof. In some embodiments, a CDR-L sequence provided herein may be combined with a variable region encoded by a variable region germline gene selected from the Vκ1, Vκ2, or Vκ3, or a variant thereof.

4. Affinity

In some embodiments, the affinity of the antibody for folate receptor alpha as indicated by K_D, is less than about 10⁻⁵ M, less than about 10⁻⁶ M, less than about 10⁻⁷ M, less than about 10⁻⁸ M, less than about 10⁻⁹ M, less than about 10⁻¹⁰ M, less than about 10⁻¹¹ M, or less than about 10⁻¹² M. In some embodiments, the affinity of the antibody is between about 10⁻⁷ M and 10⁻¹¹ M. In some embodiments, the affinity of the antibody is between about 10⁻⁷ M and 10⁻¹⁰ M. In some embodiments, the affinity of the antibody is between about 10⁻⁷ M and 10⁻⁹ M. In some embodiments, the affinity of the antibody is between about 10⁻⁷ M and 10⁻⁸ M. In some embodiments, the affinity of the antibody is between about 10⁻⁸ M and 10⁻¹¹ M. In some embodiments, the affinity of the antibody is between about 10⁻⁸ M and 10⁻¹⁰ M. In some embodiments, the affinity of the antibody is between about 10⁻⁹ M and 10⁻¹¹ M. In some embodiments, the affinity of the antibody is between about 10⁻⁹ M and 10⁻¹⁰ M.

In some embodiments, the affinity of the antibody for human folate receptor alpha, as determined by surface plasmon resonance at 25° C., and as indicated by K_D, is from about 0.36×10⁻⁹ M to about 2.21×10⁻⁹ M. In some embodiments, the affinity of the antibody for human folate receptor alpha, as determined by surface plasmon resonance at 25° C., and as indicated by K_D, is from about 8.55×10⁻¹⁰ M to about 1.70×10⁻⁸ M. In some embodiments, the affinity of the antibody for human folate receptor alpha, as determined by surface plasmon resonance at 25° C., and as indicated by K_D, is from about 5.71×10⁻¹⁰ M to about 2.58×10⁻⁸ M. In some embodiments, the affinity of the antibody for human folate receptor alpha is about any of the K_D values reported for human folate receptor alpha in the examples below.

In some embodiments the antibody has a k_a of at least about 10⁴ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of at least about 10⁵ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of at least about 10⁶ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of at least about 10⁷ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of at least about 10⁸ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of at least about 10⁹ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of between about 10⁴ M⁻¹×sec⁻¹ and about 10¹⁰ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of between about 10⁵ M⁻¹×sec⁻¹ and about 10¹⁰ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of between about 10⁶ M⁻¹×sec⁻¹ and about 10¹⁰ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a of between about 10⁷ M⁻¹×sec⁻¹ and about 10¹⁰ M⁻¹×sec⁻¹.

In some embodiments the antibody has a k_a when associating with human folate receptor alpha, as determined by surface plasmon resonance at 25° C., of from about 4.44×10⁵ M⁻¹×sec⁻¹ to about 1.61×10⁵ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a when associating with human folate receptor alpha, as determined by surface plasmon resonance at 25° C., of from about 2.90×10⁵ M⁻¹×sec⁻¹ to about 9.64×10⁹ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_a when associating with human folate receptor alpha of about any of the k_a values reported for human folate receptor alpha in the examples below.

In some embodiments the antibody has a k_d of about 10⁻⁵ sec⁻¹ or less. In some embodiments the antibody has a k_d of about 10⁻⁴ sec⁻¹ or less. In some embodiments the antibody has a k_d of about 10⁻³ sec⁻¹ or less. In some embodiments the antibody has a k_d of between about 10⁻² sec⁻¹ and about 10⁻⁵ sec⁻¹. In some embodiments the antibody has a k_d of between about 10⁻² sec⁻¹ and about 10⁻⁴ sec⁻¹. In some embodiments the antibody has a k_d of between about 10⁻³ sec⁻¹ and about 10⁻⁵ sec⁻¹.

In some embodiments the antibody has a k_d when dissociating from human folate receptor alpha, as determined by surface plasmon resonance at 25° C., of from about 8.66×10⁻⁴ sec⁻¹ to about 1.08×10⁻² sec⁻¹. In some embodiments the antibody has a k_d when dissociating from human folate receptor alpha, as determined by surface plasmon resonance at 25° C., of from about 2.28×10⁻⁴ sec⁻¹ to about 4.82×10¹ sec⁻¹. In some embodiments the antibody has a k_d when dissociating from human folate receptor alpha of about any of the k_d values reported for human folate receptor alpha in the examples below.

In some embodiments, the affinity of the antibody for cynomolgus folate receptor alpha, as determined by surface plasmon resonance at 25° C., and as indicated by K_D, is from about 0.19×10⁻⁹ M to about 2.84×10⁻⁹ M. In some embodiments, the affinity of the antibody for cynomolgus folate receptor alpha is about any of the K_D values reported for cynomolgus folate receptor alpha in the examples below.

In some embodiments, the affinity of the antibody for mouse folate receptor alpha, as determined by surface plasmon resonance at 25° C., and as indicated by K_D, is from about 0.5×10⁻⁹ M to about 9.07×10⁻⁸ M. In some embodiments, the affinity of the antibody for mouse folate receptor alpha is about any of the K_D values reported for mouse folate receptor alpha in the examples below.

In some aspects, the K_D, k_a, and k_d are determined at 25° C. In some embodiments, the K_D, k_a, and k_d are determined by surface plasmon resonance. In some embodiments, the K_D, k_a, and k_d are determined according to the methods described in the Examples provided herein.

5. Epitope Bins

In some embodiments, the antibody binds the same epitope as an antibody encompassing any of SEQ ID NOs: 308-366. In some embodiments, the antibody binds the same epitope as an antibody comprising (a) a V_H sequence comprising, consisting or, or consisting essentially of SEQ ID NOs: 308-366, and (b) a V_L sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 367-369. For example, in some embodiments, the antibody binds the same epitope as an antibody comprising any of the V_H-V_L pairs, above. In some embodiments, the antibody competes for epitope binding with an antibody encompassing any of SEQ ID NOs: 308-366. In some embodiments, the antibody competes for epitope binding with an antibody comprising (a) a V_H sequence comprising, consisting or, or consisting essentially of SEQ ID NOs: 308-366, and (b) a V_L sequence comprising, consisting of, or consisting essentially of SEQ ID NOs: 367-369. For example, in some embodiments, the antibody competes for epitope binding with an antibody comprising any of the V_H-V_L pairs, above.

6. Glycosylation Variants

In certain embodiments, an antibody may be altered to increase, decrease or eliminate the extent to which it is glycosylated. Glycosylation of polypeptides is typically either “N-linked” or “O-linked.”

“N-linked” glycosylation refers to the attachment of a carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site.

“O-linked” glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.

Addition or deletion of N-linked glycosylation sites to the antibody may be accomplished by altering the amino acid sequence such that one or more of the above-described tripeptide sequences is created or removed. Addition or deletion of O-linked glycosylation sites may be accomplished by addition, deletion, or substitution of one or more serine or threonine residues in or to (as the case may be) the sequence of an antibody.

7. Fc Variants

In certain embodiments, amino acid modifications may be introduced into the Fc region of an antibody provided herein to generate an Fc region variant. In certain embodiments, the Fc region variant possesses some, but not all, effector functions. Such antibodies may be useful, for example, in applications in which the half-life of the antibody in vivo is important, yet certain effector functions are unnecessary or deleterious. Examples of effector functions include complement-dependent cytotoxicity (CDC) and antibody-directed complement-mediated cytotoxicity (ADCC). Numerous substitutions or substitutions or deletions with altered effector function are known in the art.

In some embodiments, the Fc comprises one or more modifications in at least one of the C_H3 sequences. In some embodiments, the Fc comprises one or more modifications in at least one of the C_H2 sequences. For example, the Fc can include one or modifications selected from the group consisting of: V262E, V262D, V262K, V262R, V262S, V264S, V303R, and V305R. In some embodiments, an Fc is a single polypeptide. In some embodiments, an Fc is multiple peptides, e.g., two polypeptides. Exemplary modifications in the Fc region are described, for example, in International Patent Application No. PCT/US2017/037545, filed Jun. 14, 2017.

An alteration in in CDC and/or ADCC activity can be confirmed using in vitro and/or in vivo assays. For example, Fc receptor (FcR) binding assays can be conducted to measure FcγR binding. The primary cells for mediating ADCC, NK cells, express FcγRIII only, whereas monocytes express FcγRI, FcγRII and FcγRIII. FcR expression on hematopoietic cells is summarized in Ravetch and Kinet, Ann. Rev. Immunol., 1991, 9:457-492, incorporated by reference in its entirety.

Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest are provided in U.S. Pat. Nos. 5,500,362 and 5,821,337; Hellstrom et al., Proc. Natl. Acad. Sci. U.S.A., 1986, 83:7059-7063; Hellstrom et al., Proc. Natl. Acad. Sci. U.S.A., 1985, 82:1499-1502; and Bruggemann et al., J. Exp. Med., 1987, 166:1351-1361; each of which is incorporated by reference in its entirety. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, using an animal model such as that disclosed in Clynes et al. Proc. Natl. Acad. Sci. U.S.A., 1998, 95:652-656, incorporated by reference in its entirety.

C1q binding assays may also be carried out to confirm that the antibody is unable to bind C1q and hence lacks CDC activity. Examples of C1q binding assays include those described in WO 2006/029879 and WO 2005/100402, each of which is incorporated by reference in its entirety.

Complement activation assays include those described, for example, in Gazzano-Santoro et al., J. Immunol. Methods, 1996, 202:163-171; Cragg et al., Blood, 2003, 101:1045-1052; and Cragg and Glennie, Blood, 2004, 103:2738-2743; each of which is incorporated by reference in its entirety.

FcRn binding and in vivo clearance (half-life determination) can also be measured, for example, using the methods described in Petkova et al., Intl. Immunol., 2006, 18:1759-1769, incorporated by reference in its entirety.

8. Preparation of Antibodies

8.1. Antigen Preparation

The FOLR1 protein to be used for isolation of the antibodies may be intact FOLR1 or a fragment of FOLR1. The intact FOLR1 protein, or fragment of FOLR1, may be in the form of an isolated protein or protein expressed by a cell. Other forms of FOLR1 useful for generating antibodies will be apparent to those skilled in the art.

8.2. Monoclonal Antibodies

Monoclonal antibodies may be obtained, for example, using the hybridoma method first described by Kohler et al., Nature, 1975, 256:495-497 (incorporated by reference in its entirety), and/or by recombinant DNA methods (see e.g., U.S. Pat. No. 4,816,567, incorporated by reference in its entirety). Monoclonal antibodies may also be obtained, for example, using phage or yeast-based libraries. See e.g., U.S. Pat. Nos. 8,258,082 and 8,691,730, each of which is incorporated by reference in its entirety.

In the hybridoma method, a mouse or other appropriate host animal is immunized to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes may be immunized in vitro. Lymphocytes are then fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell. See Goding J. W., Monoclonal Antibodies: Principles and Practice3^rd ed. (1986) Academic Press, San Diego, Calif., incorporated by reference in its entirety.

The hybridoma cells are seeded and grown in a suitable culture medium that contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells.

Useful myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive media conditions, such as the presence or absence of HAT medium. Among these, preferred myeloma cell lines are murine myeloma lines, such as those derived from MOP-21 and MC-11 mouse tumors (available from the Salk Institute Cell Distribution Center, San Diego, Calif.), and SP-2 or X63-Ag8-653 cells (available from the American Type Culture Collection, Rockville, Md.). Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies. See e.g., Kozbor, J. Immunol., 1984, 133:3001, incorporated by reference in its entirety.

After the identification of hybridoma cells that produce antibodies of the desired specificity, affinity, and/or biological activity, selected clones may be subcloned by limiting dilution procedures and grown by standard methods. See Goding, supra. Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. In addition, the hybridoma cells may be grown in vivo as ascites tumors in an animal.

DNA encoding the monoclonal antibodies may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the monoclonal antibodies). Thus, the hybridoma cells can serve as a useful source of DNA encoding antibodies with the desired properties. Once isolated, the DNA may be placed into expression vectors, which are then transfected into host cells such as bacteria (e.g., E. coli), yeast (e.g., Saccharomyces or Pichia sp.), COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce antibody, to produce the monoclonal antibodies.

8.3. Humanized Antibodies

Humanized antibodies may be generated by replacing most, or all, of the structural portions of a non-human monoclonal antibody with corresponding human antibody sequences. Consequently, a hybrid molecule is generated in which only the antigen-specific variable, or CDR, is composed of non-human sequence. Methods to obtain humanized antibodies include those described in, for example, Winter and Milstein, Nature, 1991, 349:293-299; Rader et al., Proc. Nat. Acad. Sci. U.S.A., 1998, 95:8910-8915; Steinberger et al., J. Biol. Chem., 2000, 275:36073-36078; Queen et al., Proc. Natl. Acad. Sci. U.S.A., 1989, 86:10029-10033; and U.S. Pat. Nos. 5,585,089, 5,693,761, 5,693,762, and 6,180,370; each of which is incorporated by reference in its entirety.

8.4. Human Antibodies

Human antibodies can be generated by a variety of techniques known in the art, for example by using transgenic animals (e.g., humanized mice). See, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. U.S.A., 1993, 90:2551; Jakobovits et al., Nature, 1993, 362:255-258; Bruggermann et al., Year in Immuno., 1993, 7:33; and U.S. Pat. Nos. 5,591,669, 5,589,369 and 5,545,807; each of which is incorporated by reference in its entirety. Human antibodies can also be derived from phage-display libraries (see e.g., Hoogenboom et al., J. Mol. Biol., 1991, 227:381-388; Marks et al., J. Mol. Biol., 1991, 222:581-597; and U.S. Pat. Nos. 5,565,332 and 5,573,905; each of which is incorporated by reference in its entirety). Human antibodies may also be generated by in vitro activated B cells (see e.g., U.S. Pat. Nos. 5,567,610 and 5,229,275, each of which is incorporated by reference in its entirety). Human antibodies may also be derived from yeast-based libraries (see e.g., U.S. Pat. No. 8,691,730, incorporated by reference in its entirety).

9. Vectors, Host Cells, and Recombinant Methods

Embodiments are also directed to the provision of isolated nucleic acids encoding anti-FOLR1 antibodies, vectors and host cells comprising the nucleic acids, and recombinant techniques for the production of the antibodies.

For recombinant production of the antibody, the nucleic acid(s) encoding it may be isolated and inserted into a replicable vector for further cloning (i.e., amplification of the DNA) or expression. In some aspects, the nucleic acid may be produced by homologous recombination, for example as described in U.S. Pat. No. 5,204,244, incorporated by reference in its entirety.

Many different vectors are known in the art. The vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence, for example as described in U.S. Pat. No. 5,534,615, incorporated by reference in its entirety.

Illustrative examples of suitable host cells are provided below. These host cells are not meant to be limiting.

Suitable host cells include any prokaryotic (e.g., bacterial), lower eukaryotic (e.g., yeast), or higher eukaryotic (e.g., mammalian) cells. Suitable prokaryotes include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia (E. coli), Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella (S. typhimurium), Serratia (S. marcescans), Shigella, Bacilli (B. subtilis and B. licheniformis), Pseudomonas (P. aeruginosa), and Streptomyces. One useful E. coli cloning host is E. coli 294, although other strains such as E. coli B, E. coli X1776, and E. coli W3110 are suitable.

In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are also suitable cloning or expression hosts for anti-FOLR1 antibody-encoding vectors. Saccharomyces cerevisiae, or common baker's yeast, is a commonly used lower eukaryotic host microorganism. However, a number of other genera, species, and strains are available and useful, such as Spodoptera frugiperda (e.g., SF9), Schizosaccharomyces pombe, Kluyveromyces (K. lactis, K. fragilis, K. bulgaricus K. wickeramii, K. waltii, K. drosophilarum, K. thermotolerans, and K. marxianus), Yarrowia, Pichia pastoris, Candida (C. albicans), Trichoderma reesia, Neurospora crassa, Schwanniomyces (S. occidentalis), and filamentous fungi such as, for example Penicillium, Tolypocladium, and Aspergillus (A. nidulans and A. niger).

Useful mammalian host cells include COS-7 cells, HEK293 cells; baby hamster kidney (BHK) cells; Chinese hamster ovary (CHO); mouse sertoli cells; African green monkey kidney cells (VERO-76), and the like.

The host cells used to produce the anti-FOLR1 antibody of this invention may be cultured in a variety of media. Commercially available media such as, for example, Ham's F10, Minimal Essential Medium (MEM), RPMI-1640, and Dulbecco's Modified Eagle's Medium (DMEM) are suitable for culturing the host cells. In addition, any of the media described in Ham et al., Meth. Enz., 1979, 58:44; Barnes et al., Anal. Biochem., 1980, 102:255; and U.S. Pat. Nos. 4,767,704, 4,657,866, 4,927,762, 4,560,655, and 5,122,469, or WO 90/03430 and WO 87/00195 may be used. Each of the foregoing references is incorporated by reference in its entirety.

Any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics, trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art.

The culture conditions, such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.

When using recombinant techniques, the antibody can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the antibody is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. For example, Carter et al. (Bio/Technology, 1992, 10:163-167) describes a procedure for isolating antibodies which are secreted to the periplasmic space of E. coli. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min. Cell debris can be removed by centrifugation.

In some embodiments, the antibody is produced in a cell-free system. In some aspects, the cell-free system is an in vitro transcription and translation system as described in Yin et al., mAbs, 2012, 4:217-225, incorporated by reference in its entirety. In some aspects, the cell-free system utilizes a cell-free extract from a eukaryotic cell or from a prokaryotic cell. In some aspects, the prokaryotic cell is E. coli. Cell-free expression of the antibody may be useful, for example, where the antibody accumulates in a cell as an insoluble aggregate, or where yields from periplasmic expression are low. The antibodies produced in a cell-free system may be aglycosylated depending on the source of the cells.

Where the antibody is secreted into the medium, supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon® or Millipore® Pellcon® ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.

The antibody composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being a particularly useful purification technique. The suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc domain that is present in the antibody. Protein A can be used to purify antibodies that are based on human γ1, γ2, or γ4 heavy chains (Lindmark et al., J. Immunol. Meth., 1983, 62:1-13, incorporated by reference in its entirety). Protein G is useful for all mouse isotypes and for human γ3 (Guss et al., EMBO J., 1986, 5:1567-1575, incorporated by reference in its entirety).

The matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose. Where the antibody comprises a C_H3 domain, the BakerBond ABX® resin is useful for purification.

Other techniques for protein purification, such as fractionation on an ion-exchange column, ethanol precipitation, Reverse Phase HPLC, chromatography on silica, chromatography on heparin Sepharose®, chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are also available, and can be applied by one of skill in the art.

Following any preliminary purification step(s), the mixture comprising the antibody of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography using an elution buffer at a pH between about 2.5 to about 4.5, generally performed at low salt concentrations (e.g., from about 0 to about 0.25 M salt).

10. Pharmaceutical Compositions and Methods of Administration

Any of the antibodies provided herein can be provided in any appropriate pharmaceutical composition and be administered by any suitable route of administration. Suitable routes of administration include, but are not limited to, the inhalation, intraarterial, intradermal, intramuscular, intraperitoneal, intravenous, nasal, parenteral, pulmonary, and subcutaneous routes. In some embodiments, a pharmaceutical composition provided herein is administered parenterally.

The pharmaceutical composition may comprise one or more pharmaceutical excipients. Any suitable pharmaceutical excipient may be used, and one of ordinary skill in the art is capable of selecting suitable pharmaceutical excipients. Non-limiting examples of suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on a variety of factors well known in the art including, but not limited to, the way in which the dosage form will be administered to a subject and the specific antibody in the dosage form. The composition or single unit dosage form, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. Accordingly, the pharmaceutical excipients provided below are intended to be illustrative, and not limiting. Additional pharmaceutical excipients include, for example, those described in the Handbook of Pharmaceutical Excipients, Rowe et al. (Eds.) 6th Ed. (2009), incorporated by reference in its entirety.

In some embodiments, the pharmaceutical composition comprises an anti-foaming agent. Any suitable anti-foaming agent may be used. In some aspects, the anti-foaming agent is selected from an alcohol, an ether, an oil, a wax, a silicone, a surfactant, and combinations thereof. In some aspects, the anti-foaming agent is selected from a mineral oil, a vegetable oil, ethylene bis stearamide, a paraffin wax, an ester wax, a fatty alcohol wax, a long chain fatty alcohol, a fatty acid soap, a fatty acid ester, a silicon glycol, a fluorosilicone, a polyethylene glycol-polypropylene glycol copolymer, polydimethylsiloxane-silicon dioxide, ether, octyl alcohol, capryl alcohol, sorbitan trioleate, ethyl alcohol, 2-ethyl-hexanol, dimethicone, oleyl alcohol, simethicone, and combinations thereof.

In some embodiments, the pharmaceutical composition comprises a co-solvent. Illustrative examples of co-solvents include ethanol, poly(ethylene) glycol, butylene glycol, dimethylacetamide, glycerin, and propylene glycol.

In some embodiments, the pharmaceutical composition comprises a buffer. Illustrative examples of buffers include acetate, borate, carbonate, lactate, malate, phosphate, citrate, hydroxide, diethanolamine, monoethanolamine, glycine, methionine, guar gum, and monosodium glutamate.

In some embodiments, the pharmaceutical composition comprises a carrier or filler. Illustrative examples of carriers or fillers include lactose, maltodextrin, mannitol, sorbitol, chitosan, stearic acid, xanthan gum, and guar gum.

In some embodiments, the pharmaceutical composition comprises a surfactant. Illustrative examples of surfactants include d-alpha tocopherol, benzalkonium chloride, benzethonium chloride, cetrimide, cetylpyridinium chloride, docusate sodium, glyceryl behenate, glyceryl monooleate, lauric acid, macrogol 15 hydroxystearate, myristyl alcohol, phospholipids, polyoxyethylene alkyl ethers, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, polyoxylglycerides, sodium lauryl sulfate, sorbitan esters, and vitamin E polyethylene(glycol) succinate.

In some embodiments, the pharmaceutical composition comprises an anti-caking agent. Illustrative examples of anti-caking agents include calcium phosphate (tribasic), hydroxymethyl cellulose, hydroxypropyl cellulose, and magnesium oxide.

Other excipients that may be used with the pharmaceutical compositions include, for example, albumin, antioxidants, antibacterial agents, antifungal agents, bioabsorbable polymers, chelating agents, controlled release agents, diluents, dispersing agents, dissolution enhancers, emulsifying agents, gelling agents, ointment bases, penetration enhancers, preservatives, solubilizing agents, solvents, stabilizing agents, and sugars. Specific examples of each of these agents are described, for example, in the Handbook of Pharmaceutical Excipients, Rowe et al. (Eds.) 6th Ed. (2009), The Pharmaceutical Press, incorporated by reference in its entirety.

In some embodiments, the pharmaceutical composition comprises a solvent. In some aspects, the solvent is saline solution, such as a sterile isotonic saline solution or dextrose solution. In some aspects, the solvent is water for injection.

In some embodiments, the pharmaceutical compositions are in a particulate form, such as a microparticle or a nanoparticle. Microparticles and nanoparticles may be formed from any suitable material, such as a polymer or a lipid. In some aspects, the microparticles or nanoparticles are micelles, liposomes, or polymersomes.

Further provided herein are anhydrous pharmaceutical compositions and dosage forms comprising an antibody, since water can facilitate the degradation of some antibodies.

Anhydrous pharmaceutical compositions and dosage forms provided herein can be prepared using anhydrous or low moisture containing ingredients and low moisture or low humidity conditions. Pharmaceutical compositions and dosage forms that comprise lactose and at least one active ingredient that comprises a primary or secondary amine can be anhydrous if substantial contact with moisture and/or humidity during manufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition can be prepared and stored such that its anhydrous nature is maintained. Accordingly, anhydrous compositions can be packaged using materials known to prevent exposure to water such that they can be included in suitable formulary kits. Examples of suitable packaging include, but are not limited to, hermetically sealed foils, plastics, unit dose containers (e.g., vials), blister packs, and strip packs.

In some embodiments lactose-free compositions are provided herein which comprise excipients that are well known in the art and are listed, for example, in the U.S. Pharmocopia (USP) SP (XXI)/NF (XVI). In general, lactose-free compositions comprise an active ingredient, a binder/filler, and a lubricant in pharmaceutically compatible and pharmaceutically acceptable amounts. Exemplary lactose-free dosage forms comprise an active ingredient, microcrystalline cellulose, pre gelatinized starch, and magnesium stearate.

Also provided are pharmaceutical compositions and dosage forms that comprise one or more excipients that reduce the rate by which an antibody will decompose. Such excipients, which are referred to herein as “stabilizers,” include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers.

10.1. Parenteral Dosage Forms

In certain embodiments, provided are parenteral dosage forms. Parenteral dosage forms can be administered to subjects by various routes including, but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular, and intraarterial. Because their administration typically bypasses subjects' natural defenses against contaminants, parenteral dosage forms are typically, sterile or capable of being sterilized prior to administration to a subject. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms are well known to those skilled in the art. Examples include, but are not limited to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

Excipients that increase the solubility of one or more of the antibodies disclosed herein can also be incorporated into the parenteral dosage forms.

10.2. Dosage and Unit Dosage Forms

In human therapeutics, the doctor will determine the posology which he considers most appropriate according to a preventive or curative treatment and according to the age, weight, condition and other factors specific to the subject to be treated.

In certain embodiments, a composition provided herein is a pharmaceutical composition or a single unit dosage form. Pharmaceutical compositions and single unit dosage forms provided herein comprise a prophylactically or therapeutically effective amount of one or more prophylactic or therapeutic antibodies.

The amount of the antibody or composition which will be effective in the prevention or treatment of a disorder or one or more symptoms thereof will vary with the nature and severity of the disease or condition, and the route by which the antibody is administered. The frequency and dosage will also vary according to factors specific for each subject depending on the specific therapy (e.g., therapeutic or prophylactic agents) administered, the severity of the disorder, disease, or condition, the route of administration, as well as age, body, weight, response, and the past medical history of the subject. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

In certain embodiments, exemplary doses of a composition include milligram or microgram amounts of the antibody per kilogram of subject or sample weight (e.g., about 10 micrograms per kilogram to about 50 milligrams per kilogram, about 100 micrograms per kilogram to about 25 milligrams per kilogram, or about 100 microgram per kilogram to about 10 milligrams per kilogram). In certain embodiment, the dosage of the antibody provided herein, based on weight of the antibody, administered to prevent, treat, manage, or ameliorate a disorder, or one or more symptoms thereof in a subject is 0.1 mg/kg, 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 10 mg/kg, or 15 mg/kg or more of a subject's body weight. In another embodiment, the dosage of the composition or a composition provided herein administered to prevent, treat, manage, or ameliorate a disorder, or one or more symptoms thereof in a subject is 0.1 mg to 200 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 0.1 mg to 20 mg, 0.1 mg to 15 mg, 0.1 mg to 10 mg, 0.1 mg to 7.5 mg, 0.1 mg to 5 mg, 0.1 to 2.5 mg, 0.25 mg to 20 mg, 0.25 to 15 mg, 0.25 to 12 mg, 0.25 to 10 mg, 0.25 mg to 7.5 mg, 0.25 mg to 5 mg, 0.25 mg to 2.5 mg, 0.5 mg to 20 mg, 0.5 to 15 mg, 0.5 to 12 mg, 0.5 to 10 mg, 0.5 mg to 7.5 mg, 0.5 mg to 5 mg, 0.5 mg to 2.5 mg, 1 mg to 20 mg, 1 mg to 15 mg, 1 mg to 12 mg, 1 mg to 10 mg, 1 mg to 7.5 mg, 1 mg to 5 mg, or 1 mg to 2.5 mg.

The dose can be administered according to a suitable schedule, for example, once, two times, three times, or for times weekly. It may be necessary to use dosages of the antibody outside the ranges disclosed herein in some cases, as will be apparent to those of ordinary skill in the art. Furthermore, it is noted that the clinician or treating physician will know how and when to interrupt, adjust, or terminate therapy in conjunction with subject response.

Different therapeutically effective amounts may be applicable for different diseases and conditions, as will be readily known by those of ordinary skill in the art. Similarly, amounts sufficient to prevent, manage, treat or ameliorate such disorders, but insufficient to cause, or sufficient to reduce, adverse effects associated with the antibodies provided herein are also encompassed by the herein described dosage amounts and dose frequency schedules. Further, when a subject is administered multiple dosages of a composition provided herein, not all of the dosages need be the same. For example, the dosage administered to the subject may be increased to improve the prophylactic or therapeutic effect of the composition or it may be decreased to reduce one or more side effects that a particular subject is experiencing.

In certain embodiments, treatment or prevention can be initiated with one or more loading doses of an antibody or composition provided herein followed by one or more maintenance doses.

In certain embodiments, a dose of an antibody or composition provided herein can be administered to achieve a steady-state concentration of the antibody in blood or serum of the subject. The steady-state concentration can be determined by measurement according to techniques available to those of skill or can be based on the physical characteristics of the subject such as height, weight and age.

In certain embodiments, administration of the same composition may be repeated and the administrations may be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months. In other embodiments, administration of the same prophylactic or therapeutic agent may be repeated and the administration may be separated by at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, or 6 months.

11. Therapeutic Applications

For therapeutic applications, the antibodies of the invention are administered to a mammal, generally a human, in a pharmaceutically acceptable dosage form such as those known in the art and those discussed above. For example, the antibodies of the invention may be administered to a human intravenously as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intra-cerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, or intratumoral routes. The antibodies also are suitably administered by peritumoral, intralesional, or perilesional routes, to exert local as well as systemic therapeutic effects. The intraperitoneal route may be particularly useful, for example, in the treatment of ovarian tumors.

The antibodies provided herein may be useful for the treatment of any disease or condition involving folate receptor alpha (FOLR1). In some embodiments, the disease or condition is a disease or condition that can be diagnosed by overexpression of folate receptor alpha. In some embodiments, the disease or condition is a disease or condition that can benefit from treatment with an anti-folate receptor alpha antibody. In some embodiments, the disease or condition is a cancer.

Any suitable cancer may be treated with the antibodies provided herein. Illustrative suitable cancers include, for example, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, brain tumor, bile duct cancer, bladder cancer, bone cancer, breast cancer (including triple-negative breast cancer, or TNBC), bronchial tumor, carcinoma of unknown primary origin, cardiac tumor, cervical cancer, chordoma, colon cancer, colorectal cancer, craniopharyngioma, ductal carcinoma, embryonal tumor, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, fallopian tube carcinoma, fibrous histiocytoma, Ewing sarcoma, eye cancer, germ cell tumor, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, gestational trophoblastic disease, glioma, head and neck cancer, hepatocellular cancer, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, Kaposi sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ, lung cancer, macroglobulinemia, malignant fibrous histiocytoma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer with occult primary, midline tract carcinoma involving NUT gene, mouth cancer, multiple endocrine neoplasia syndrome, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic/myeloproliferative neoplasm, nasal cavity and par nasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer (NSCLC), oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytomas, pituitary tumor, pleuropulmonary blastoma, primary central nervous system lymphoma, primary peritoneal carcinoma, prostate cancer, rectal cancer, renal cell cancer, renal pelvis and ureter cancer, retinoblastoma, rhabdoid tumor, salivary gland cancer, Sezary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, spinal cord tumor, stomach cancer, T-cell lymphoma, teratoid tumor, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, and Wilms tumor.

In some embodiments, the disease to be treated with the antibodies provided herein is gastric cancer, colorectal cancer, renal cell carcinoma, cervical cancer, non-small cell lung carcinoma, ovarian cancer, uterine cancer, fallopian tube carcinoma, primary peritoneal carcinoma, uterine corpus carcinoma, endometrial carcinoma, prostate cancer, breast cancer, head and neck cancer, brain carcinoma, liver cancer, pancreatic cancer, mesothelioma, and/or a cancer of epithelial origin. In particular embodiments, the disease is colorectal cancer. In some embodiments, the disease is ovarian cancer. In some embodiments, the disease is breast cancer. In some embodiments, the disease is triple-negative breast cancer (TNBC). In some embodiments, the disease is lung cancer. In some embodiments, the disease is non-small cell lung cancer (NSCLC). In some embodiments, the disease is head and neck cancer. In some embodiments, the disease is renal cell carcinoma. In some embodiments, the disease is brain carcinoma. In some embodiments, the disease is endometrial cancer.

12. Diagnostic Applications

In some embodiments, the antibodies provided herein are used in diagnostic applications. For example, an anti-FOLR1 antibody may be useful in assays for FOLR1 protein. In some aspects the antibody can be used to detect the expression of FOLR1 in various cells and tissues. These assays may be useful, for example, in making a diagnosis and/or prognosis for a disease, such as a cancer.

In some diagnostic and prognostic applications, the antibody may be labeled with a detectable moiety. Suitable detectable moieties include, but are not limited to radioisotopes, fluorescent labels, and enzyme-substrate labels. In another embodiment, the anti-FOLR1 antibody need not be labeled, and the presence of the antibody can be detected using a labeled antibody which specifically binds to the anti-FOLR1 antibody.

13. Affinity Purification Reagents

The antibodies of the invention may be used as affinity purification agents. In this process, the antibodies may be immobilized on a solid phase such a resin or filter paper, using methods well known in the art. The immobilized antibody is contacted with a sample containing the folate receptor alpha protein (or fragment thereof) to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the folate receptor alpha protein, which is bound to the immobilized antibody. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0 that will release the folate receptor alpha protein from the antibody.

14. Kits

In some embodiments, an anti-FOLR1 antibody provided herein is provided in the form of a kit, i.e., a packaged combination of reagents in predetermined amounts with instructions for performing a procedure. In some embodiments, the procedure is a diagnostic assay. In other embodiments, the procedure is a therapeutic procedure.

In some embodiments, the kit further comprises a solvent for the reconstitution of the anti-FOLR1 antibody. In some embodiments, the anti-FOLR1 antibody is provided in the form of a pharmaceutical composition.

EXAMPLES

Example 1

Generation and Primary Screening of Anti-FOLR1 Antibodies

Antibody Fab libraries were constructed using a standard overlap extension PCR protocol with mutagenic primers targeting complementary determining regions (CDRs). See Heckman and Pease, Nat. Protoc., 2007, 2:924-932; Stafford et al., 2014, Protein Eng. Des. Sel. 27:97-109, both incorporated by reference in their entireties. Selections for novel antibodies were performed using standard ribosome display protocols. See Dreier and Plückthun, 2011, Methods Mol Biol 687:283-306, which is incorporated herein by reference in its entirety.

Initial antibody leads from ribosome display were derived from a naïve human library which was constructed by overlapping PCR using trastuzumab HC as the base template. CDRs H1 and H2 were randomized with the same design as described by Lee et al., J. Mol. Biol. 2004, 340:1073-1093 using oligonucleotides purchased from Integrated DNA Technologies. In this design, CDRs H1 and H2 closely match the observed amino acid distributions of natural human antibodies. CDR H3 was diversified using oligonucleotides incorporating trimer phosphoramidite mixtures (TRIMs) for amino acid randomization. The TRIM oligos were synthesized as described by Yagodkin A et al., Nucleosides Nucleotides Nucleic Acids 2007, 26:473-97. Specifically, six separate oligonucleotides containing TRIMs were used to make 6 separate H3 loop-lengths (13-18; as defined by Zemlin et al.) to match the most common loop lengths observed in the human repertoire. Together these loop lengths comprise approximately 54.5% of the naturally-occurring loop length variation in human IgGs as reported by Zemlin et al., J. Mol. Biol. 2003, 334:733-749. The frequency distribution of each amino acid was designed to closely match the observed distribution of amino acids in CDR H3 of human IgGs as reported by Zemlin et al. Altogether, the library closely matches natural human antibody variation which is known in the field to improve antibody stability and folding of antibodies as described by Zhai et al., J Mol Biol. 2011, 412:55-71. The heavy chain (HC) library was paired with a constant, unmodified trastuzumab light chain (LC) throughout the selection process as described by Stafford et al., Protein Eng Des Sel 2014, 27:97-109.

Affinity maturated antibody leads (e.g., SRP 1848 antibodies, below) were derived from a focused library, biased towards two leads, which was constructed by overlapping PCR using “soft-randomized” oligonucleotides purchased from Eurofins MWG Operon. Soft-randomization is a process in which a biased distribution of nucleotides is used for each soft-randomized codon such that the parent amino acid sequence is coded more frequently than other amino acids-30% of the time. Other amino acids are coded at each position but at a lower percentage. At each soft-randomized position, 70% of the parent nucleotide is mixed with 10% of the other three nucleotides. For the library, CDRs H1, H2, and H3 were soft-randomized simultaneously and selected by standard ribosome display protocols. As with the selection of initial leads, the affinity matured antibodies were paired with a constant, unmodified trastuzumab LC throughout the selection process as described by Stafford et al., Protein Eng Des Sel 2014, 27:97-109.

Selections for novel antibodies were performed using standard ribosome display protocols. See Dreier and Plückthun, Methods Mol. Biol., 2003, 687:283-306, Clifton, N.J., incorporated by reference in its entirety. Fab ribosome display selections were performed according to published protocols. See Stafford et al., 2014, Protein Eng. Des. Sel. 27:97-109; Hanes and Plückthun, Proc. Natl. Acad. Sci. U.S.A., 1997, 94:4937-4942; both incorporated by reference in their entireties. After multiple rounds of selection, the DNA from RT-PCR output was cloned into an optimized vector for cell-free expression using standard molecular biology techniques. See Yin et al., mAbs, 2012, 4:217-225, incorporated by reference in its entirety. All constructs were HIS- and FLAG-tagged to streamline purification and testing during screening.

Libraries of antibody variants generated by selection workflow were transformed into E. coli and grown on agar plates with antibiotic (kanamycin). Individual colonies were grown in liquid broth (TB+kanamycin), and used as a template for DNA amplification via rolling circle amplification (RCA). The variants were then expressed in cell-free protein synthesis reactions as described in Yin et al., mAbs, 2012, 4:217-225.

Briefly, cell-free extracts were treated with 50 μM iodoacetamide for 30 min at room temperature (20° C.) and added to a premix containing cell-free components (see Cai et al., Biotechnol Prg, 2015, 3:823-831, incorporated by reference in its entirety) and 10% (v/v) RCA DNA template (approximately 10 μg/mL DNA) for HC variants, in addition to 2.5 μg/mL Trastuzumab LC which is present for antibody assembly but is not varied in the library. Sixty microliters of cell-free reactions were incubated at 30° C. for 12 hr on a shaker at 650 rpm in 96-well plates. Four hundred to one-thousand-five-hundred (400 to 1500) colonies were screened, depending on the predicted diversity of different selection campaigns.

Following synthesis, each reaction was diluted 1:50 into PBS (pH 7.4) with 3% fetal bovine serum (FBS), and expressed variants were tested for functional activity via cell-based ELISA binding to CHO-hFOLR1 cells (human FOLR1 expressed recombinantly in Chinese Hamster Ovary cells). Briefly, 384-well plates were seeded with CHO-control or CHO-hFOLR1 cells the day before the assay. On the day of the assay, cells were fixed with 20 uL of 4% paraformaldehyde in PBS for 15 minutes in the dark, washed with PBS, and then blocked with 30% FBS in PBS for 30 minutes at room temperature. Antibody variants of interest (1:50 diluted cell-free reaction) were allowed to bind to the fixed CHO-hFOLR1 cells, and detected with secondary antibodies (e.g. HRP-conjugated Anti-human Fc or anti-FLAG) and then detected with chemiluminescent substrate (Pierce ELISA SuperSignal™ Substrate). Chemiluminescence was quantified on a Molecular Devices SpectraMax® M5 plate reader. Top hits were selected based on cell-based ELISA signal/noise ratio, and their nucleotides were sequenced. Based on binding activity and sequence analysis, a subset of variants was selected for further scale-up and characterization.

The top leads from ELISA-based screening were cultured, and plasmid minipreps were performed using a QIAprep® 96 Turbo miniprep kit (Qiagen) according to the manufacturer's instructions. 10 μg/mL miniprepped DNA was added to 4 mL cell-free reactions and incubated overnight for 12 hr at 30° C., at 650 rpm. In the case of IgG variants with a common Trastuzumab LC, 7.5 ug/mL of the HC variant DNA and 2.5 ug/mL of the common Trastuzumab LC were added to the reaction.

Expressed variants from clarified cell-free reactions were purified via immobilized metal ion affinity chromatography (IMAC) purification using a semi-automated high throughput batch purification method. Briefly, purifications were performed in a 96-well plate format where 50 jiL/well of IMAC resin (Ni Sepharose High Performance, GE Healthcare) was equilibrated in IMAC binding buffer (50 mM Tris pH 8.0, 300 mM NaCl, 10 mM imidazole), incubated with 1 mL cell-free reaction for 15 minutes followed by two washes in IMAC binding buffer. His-tagged antibody variants were then eluted using 200 μL IMAC elution buffer (50 mM Tris pH 8.0, 300 mM NaCl, 500 mM imidazole) and buffer exchanged into PBS using a 96-well Zeba plate (7 kD MWCO, Thermo Fisher). Purified antibodies were quantified via high throughput capillary electrophoresis using the LabChip GXII (Perkin Elmer) against a Herceptin standard curve, according to the manufacturer's instructions.

Exemplary affinity-matured antibodies are reported in Table 5, below.

TABLE 5
Affinity Matured (SRP1848) Antibodies
		SEQ ID		SEQ ID
Antibody	VH	NO.	VL	NO.
1	SRP1848-A01	308	Trastuzumab	367
2	SRP1848-A02	309	Trastuzumab	367
3	SRP1848-A04	310	Trastuzumab	367
4	SRP1848-A06	311	Trastuzumab	367
5	SRP1848-A07	312	Trastuzumab	367
6	SRP1848-A08	313	Trastuzumab	367
7	SRP1848-A09	314	Trastuzumab	367
8	SRP1848-A10	315	Trastuzumab	367
9	SRP1848-B01	316	Trastuzumab	367
10	SRP1848-B03	317	Trastuzumab	367
11	SRP1848-B04	318	Trastuzumab	367
12	SRP1848-B05	319	Trastuzumab	367
13	SRP1848-B06	320	Trastuzumab	367
14	SRP1848-B07	321	Trastuzumab	367
15	SRP1848-B09	322	Trastuzumab	367
16	SRP1848-B10	323	Trastuzumab	367
17	SRP1848-B11	324	Trastuzumab	367
18	SRP1848-C01	325	Trastuzumab	367
19	SRP1848-C03	326	Trastuzumab	367
20	SRP1848-C04	327	Trastuzumab	367
21	SRP1848-C05	328	Trastuzumab	367
22	SRP1848-C07	329	Trastuzumab	367
23	SRP1848-C10	330	Trastuzumab	367
24	SRP1848-D02	331	Trastuzumab	367
25	SRP1848-D03	332	Trastuzumab	367
26	SRP1848-D04	333	Trastuzumab	367
27	SRP1848-D05	334	Trastuzumab	367
28	SRP1848-D07	335	Trastuzumab	367
29	SRP1848-D09	336	Trastuzumab	367
30	SRP1848-D10	337	Trastuzumab	367
31	SRP1848-E01	338	Trastuzumab	367
32	SRP1848-E02	339	Trastuzumab	367
33	SRP1848-E03	340	Trastuzumab	367
34	SRP1848-E05	341	Trastuzumab	367
35	SRP1848-E06	342	Trastuzumab	367
36	SRP1848-E07	343	Trastuzumab	367
37	SRP1848-F01	344	Trastuzumab	367
38	SRP1848-F02	345	Trastuzumab	367
39	SRP1848-F04	346	Trastuzumab	367
40	SRP1848-F05	347	Trastuzumab	367
41	SRP1848-F06	348	Trastuzumab	367
42	SRP1848-F07	349	Trastuzumab	367
43	SRP1848-F08	350	Trastuzumab	367
44	SRP1848-F09	351	Trastuzumab	367
45	SRP1848-F10	352	Trastuzumab	367
46	SRP1848-F11	353	Trastuzumab	367
47	SRP1848-G01	354	Trastuzumab	367
48	SRP1848-G03	355	Trastuzumab	367
49	SRP1848-G04	356	Trastuzumab	367
50	SRP1848-G06	357	Trastuzumab	367
51	SRP1848-G07	358	Trastuzumab	367
52	SRP1848-G09	359	Trastuzumab	367
53	SRP1848-G10	360	Trastuzumab	367
54	SRP1848-G11	361	Trastuzumab	367
55	SRP1848-H01	362	Trastuzumab	367

Example 2

Preparation of SCFVS

A single-chain antibody is made in either the V_HV_L or V_LV_H orientation with a linker sequence between the V_H and V_L domains. Typically scFv linkers are composed of (GGGGS)n repeats where n=3, 4, 5, or 6 for linkers of 15, 20, 25, or 30 residues respectively. For cell-free expression, an N-terminal Met is added, but for mammalian expression a leader peptide is added. On the C-terminal end of the scFv, an Fc sequence can be added to extend in vivo half-life or the scFv can be used directly. An optional linker sequence can be incorporated between the scFv and the Fc. An exemplary scFv-Fc linker sequence is AAGSDQEPKSS (SEQ ID NO: 378). C-terminal affinity tags can optionally be added to facilitate purification and assay development. An exemplary affinity tag is a C-terminal FlagHis tag GSGDYKDDDDKGSGHHHHHH (SEQ ID NO: 376). A stop codon is typically inserted at the end of the sequence. An exemplary scFv can include an N-terminal Met residue, a V_H domain, a GGGGSGGGGSGGGGS (SEQ ID NO: 377) linker, a V_L domain, an AAGSDQEPKSS (SEQ ID NO: 378) linker, an Fc domain, a FlagHis tag, and a stop codon.

Example 3

Affinity and Kinetic Binding Analyses

Anti-Fc polyclonal antibodies were immobilized onto a CM5 chip (GE Life Sciences) using amine coupling chemistry (from Amine Coupling Kit, GE Life Sciences). The immobilization steps were carried out at a flow rate of 25 μL/min in 1×HBS-EP+buffer (GE Life Sciences; 10× Stock diluted before use). The sensor surfaces were activated for 7 min with a mixture of N-hydroxysuccinimide (NHS, 0.05 M) and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC, 0.2 M). The anti-Fc polyclonal antibodies were injected over all 4 flow cells at a concentration of 25 μg/mL in 10 mM sodium acetate, pH 4.5, for 7 min. Ethanolamine (1 M, pH 8.5) was injected for 7 min to block any remaining activated groups. An average of 12,000 response units (RU) of capture antibody was immobilized on each flow cell.

Off-rate and kinetic binding experiments were performed at 25° C. using 1×HBS-EP+buffer. Test and control antibodies were injected over the anti-Fc surface at concentrations of 5-10 μg/mL for 12 seconds at a flow rate of 10 μL/min on flow cells 2, 3 and 4, followed by a buffer wash for 30 seconds at the same flow rate. Kinetic characterization of antibody samples was carried out with a single concentration of antigen (for off-rate ranking) or a 1:2 dilution series of antigen (for kinetic characterization) and 1 injection of 0 nM antigen. After capturing ligand (antibody) on the anti-Fc surface, the analyte (human FOLR1-HIS) was bound at 50, 25, 12.5, 6.25 and 0 nM for 180 seconds, followed by a 600 second dissociation phase at a flow rate of 50 μL/min. Between each ligand capture and analyte binding cycle, regeneration was carried out using 2 injections of 10 mM glycine pH 2.0 for 30 seconds at 30 L/min, followed by a 30 second buffer wash step.

The data were fit with the Biacore T200 Evaluation software, using a 1:1 Langmuir binding model. K_D (affinity, nM) was determined as a ratio of the kinetic rate constants calculated from the fits of the association and dissociation phases.

Example 4

Flow Cytometry-Based Cell Binding Assay

Variants with expression levels >250 nM were tested in a fluorescence-activated cell sorting (FACS) cell-binding assay. CHO cells were transfected to stably express human (CHO-hFOLR1), cynomolgus (CHO-cFOLR1), or mouse (CHO-mFOLR1) target molecule FOLR1 on the cell surface. Parental CHO cells were used as a negative control to determine background binding levels. Parental CHO and stably transfected CHO-hFOLR1, CHO-cFOLR1, and CHO-mFOLR1 cells were cultured in Ham's F-12: high glucose DMEM (50:50) (Corning, Cellgro-Mediatech) supplemented with 10% heat-inactivated fetal bovine serum (Corning, Cellgro-Mediatech), 1% Penicillin/Streptomycin (Corning, Cellgro-Mediatech) and 2 mmol/L-glutamax (Life Technology).

A mixture of fluoresecent-labeled parental CHO cells and unlabeled CHO-hFOLRlcells were prepared as follows. Parental CHO cells were washed twice in PBS and incubated in PBS containing with 1 nM CellTrace™ Oregon Green488® (Life Technologies) at 37° C. for 30 minutes. Labeled parental CHO cells were then washed 2× with Ham's F-12 media and 2× with FACS buffer (PBS with 1% bovine serum albumin). Unlabeled CHO-hFOLR1 cells were similarly washed and prepared. Labeled parental CHO and unlabeled CHO-hFOLR1 cells were combined at 1:1 ratio and seeded at 50 μL per well (200,000 cells per well) in 96 well polypropylene plates. Cells were mixed with 50 μL of test antibodies (i.e., anti-FOLR1 variants) serially diluted in FACS buffer and incubated on ice for 60 mins. Cells were washed with FACS buffer and incubated on ice for 60 mins with 100 μL FACS buffer containing 2.5 μg/mL R-Phycoerythrin-conjugated goat anti-Human IgG (Jackson ImmunoResearch Laboratories, West Grove, Pa.). Cells were washed twice with FACS buffer, fixed in 2% paraformaldehyde in PBS (Santa Cruz Biotechnology; Dallas, Tex.) for 10 mins on ice in the dark, and analyzed using the BD LSR II Flow Cytometer (BD Biosciences; San Jose, Calif.). Data were analyzed using FlowJo® software (FlowJo, LLC; Ashland, Oreg.) to determine mean fluorescence intensities. Binding constants were calculated using the statistical software, GraphPad Prism (GraphPad Software; La Jolla, Calif.) using the nonlinear regression equation, one site-specific binding with Hill slope. Secondary antibody alone was used as a control, in addition to measuring non-specific antibody binding to CHO parental cells.

This procedure was repeated to assess cell binding in CHO-cFOLR1 and CHO-mFOLR1 cells.

Example 5

Cell-Killing Analysis

The internalization of the antibodies was evaluated by a secondary antibody cell killing assay on target positive cells. FOLR1-positive KB cells were obtained from ATCC, and FOLR1-positive Igrovl cells were obtained from NIH. The cells were maintained in Ham's F-12: high glucose DMEM (50:50) (Corning, Cellgro-Mediatech) supplemented with 10% heat-inactivated fetal bovine serum (Corning, Cellgro-Mediatech, Manassas, Va.), 1% Penicillin/Streptomycin (Corning, Cellgro-Mediatech, Manassas, Va.) and 2 mmol/L-glutamax (Thermo Fisher Scientific, Waltham, Mass.). Adherent cells were washed twice with calcium and magnesium-free Hanks Balanced Salt Solution (HBSS), harvested with HYQ®TASE™ (Hyclone; Thermo Fisher Scientific, Waltham, Mass.) and counted by the Vi-CELL Cell Viability Analyzers (Beckman Coulter, Indianapolis, Ind.). A total of 625 cells were seeded in each well of a 384-well flat bottom white polystyrene plate. Lead antibodies were formulated at 4-fold starting concentration in the cell culture medium and filtered through MultiScreenHTS 96-Well Filter Plates (Millipore; Billerica, Mass.). Serial dilutions of test antibody (1:3 serial dilution starting from 200 nM) was added into treatment wells, and an anti-human Fc nanobody conjugated to hemiasterlin via a cleavable linker was then added into each well at a fixed final concentration of 20 nM. Assay plates were cultured at 37° C. in a CO₂ incubator for 120 hrs before assay. For cell viability measurement, 30 μL of Cell Titer-Glo® reagent (Promega Corp. Madison, Wis.) was added into each well, and plates were processed as per product instructions. Relative luminescence was measured on an ENVISION® plate reader (Perkin-Elmer; Waltham, Mass.). Relative luminescence readings were converted to percent viability using untreated cells as controls. Data was fitted with nonlinear regression analysis, using a log(inhibitor) vs. response-variable slope, 4 parameter fit with GraphPad Prism (GraphPad v 5.0, Software; San Diego, Calif.). Data was expressed as relative cell viability (ATP content) % vs. dose of antibody.

Example 6

Generation of Hybridoma

Immunocompetent mice (C57BL/6) were immunized with mouse MC38 cells overexpressing human FOLR1. FOLR1-specific antibodies were detected in the sera, and the spleen was harvested and fused with P3X cells to generate the hybridomas (Aragen Biosciences, Morgan Hill, Calif.), similar to what has been previously described. See Chronopoulou, et al., 2014, Methods Mol Biol 1131:47-70, and Kim, et al., 2014, Methods Mol Biol 1131:31-45, each of which is incorporated herein by reference in its entirety. Total RNA was extracted from hybridoma cells using QIAGEN RNeasy Mini Kit (Cat No. 74104) and converted to cDNA using a Clontech SMARTer RACE cDNA Amplification Kit (Cat. No. 634923) (Lake Pharma, Belmont, Calif.). Positive clones were identified by gel electrophoresis, cloned using an Invitrogen TOPO kit, and sequenced using standard Sanger methods. The CDRs for m6D1 were grafted onto human antibody frameworks V_H1-18, V_H3-33, V_H2-5, V_H2-70, V_H4-30-4, Vk1-5, Vk3-11, Vk2-30, Vk1-33, and Vk1-16 by standard methodology to yield humanized antibodies. See Kuramochi, et al., 2014, Methods Mol Biol 1060:123-137, which is incorporated herein by reference in its entirety. Of these grafts, the h6D1-HC3/LC4 (V_H3-33/Vk3-11 grafts) and h6D1-HC3/LC5 (V_H3-33/Vk1-5 grafts) IgGs gave the best yield when expressed in cell-free and maintained the highest affinity. Both HC3/LC4 and HC3/LC5 humanized variants were progressed into affinity maturation by Fab-based ribosome display (as described above) targeting the heavy chain CDRs by soft-randomization leaving the light-chain constant, as described in Stafford, et al., 2014, Protein Eng Des Sel 4:97-109, which is incorporated herein by reference in its entirety.

Certain antibodies were generated by affinity maturation of humanized mouse antibodies. Exemplary antibody candidates are reported in Table 6, below.

TABLE 6
Affinity-matured humanized antibodies (SRP2060).
		SEQ ID		SEQ ID
Antibody	VH	NO.	VL	NO.
56	SRP2060-E10	363	H6D1-LC4	368
57	SRP2060-E05	364	H6D1-LC4	368
58	SRP2060-B01	365	H6D1-LC5	369
59	SRP2060-A06	366	H6D1-LC5	369

Example 7

Characteristics of Illustrative Anti-FOLR1 Antibodies

Tables 7 through 9 show results obtained using the illustrative antibodies described herein.

Table 7 shows results obtained with antibodies isolated from affinity-maturation of initial antibody leads obtained from a naïve Fab TRiM ribosome display library, constructed on a Trastuzumab heavy chain (HC) framework.

Table 8 shows kinetic binding results obtained for the same antibodies listed in Table 7.

Table 9 shows results obtained from antibodies isolated from humanized mouse clone candidates.

TABLE 7
Affinity-matured antibodies from initial leads (Trastuzumab HC framework).
	KB, 2°	Igrov, 2°
	Antibody	Antibody
	Cell	Cell	CHO-
	Killing,	Killing,	human	CHO-cyno	CHO-mouse
	Nb-239	Nb-SC239	FoIR1	FoIR1	FoIR1
	EC50	Span	EC50	Span	Bmax	KD	Bmax	KD	Bmax	KD
Fab-HC Variant ID	(nM)	(%)	(nM)	(%)	(MFI)	(nM)	(MFI)	(nM)	(MFI)	(nM)
SRP1848-A01	0.064	94	0.015	71	25899	0.39	22552	0.44	16475	0.8
SRP1848-A02	0.028	94	0.039	71	24710	0.64	18500	0.50	18569	2.4
SRP1848-A07	0.062	95	0.029	68	29182	0.61	23643	0.43	9646	0.9
SRP1848-C03	0.074	93	0.035	72	29143	0.51	25148	0.50	3310	3.0
SRP1848-F04	0.096	93	0.015	73	26867	0.73	26353	0.55	2741	11.0
SRP1848-B04	0.035	94	0.018	70	27818	0.72	27796	0.65	2187	17.9
SRP1848-B11	0.058	93	0.026	74	28394	0.56	22885	0.34	1632	3.9
SRP1848-F07	0.057	92	0.018	71	27371	0.58	18662	0.56	1387	8.8
SRP1848-E06	0.060	93	0.025	74	25611	0.48	15755	0.26	2349	1.2
SRP1848-A09	0.060	93	0.026	71	28910	0.61	20248	0.31	7990	1.0
SRP1848-E07	0.059	94	0.013	73	27284	0.54	20381	0.23	11837	1.2
SRP1848-G03	0.064	91	0.021	76	26424	0.82	19238	0.44	2220	2.4
SRP1848-A04	0.052	92	0.015	64	26810	0.43	23055	0.30	3888	2.0
SRP1848-H01	0.049	96	0.016	67	26985	0.59	17227	0.28	3950	33.8
SRP1848-B10	0.040	97	0.020	71	28186	0.83	21268	0.44	2455	7.3
SRP1848-C07	0.065	93	0.013	67	28757	0.62	18136	0.23	3170	1.4
SRP1848-F05	0.061	94	0.015	74	27155	0.72	24731	0.61	5100	18.0
SRP1848-D02	0.034	93	0.027	71	28804	0.60	27973	0.61	916	87.0
SRP1848-A08	0.039	93	0.013	65	28554	0.62	26197	0.45	3202	2.5
SRP1848-E03	0.057	94	0.027	73	26694	0.76	17427	0.43	5939	0.5
SRP1848-A10	0.033	96	0.027	75	27097	0.66	14816	0.47	10167	1.2
SRP1848-F10	0.038	94	0.009	68	25554	0.36	20700	0.40	1742	6.9
SRP1848-D05	0.055	92	0.030	73	26748	0.57	22202	0.45	1360	14.0
SRP1848-C01	0.060	90	0.023	68	28527	0.66	25941	0.60	1369	26.0
SRP1848-F01	0.047	91	0.018	69	25240	0.56	21491	0.43	3750	1.8
SRP1848-D04	0.380	97	0.068	77	29297	2.21	25737	2.84	NB	NB
SRP1848-E05	0.071	95	0.027	78	27306	0.46	28170	0.55	NB	NB
SRP1848-A06	0.046	93	0.020	72	24521	0.47	20170	0.30	2767	2.4
SRP1848-B01	0.064	95	0.031	82	26634	1.06	23881	0.83	3404	16.4
SRP1848-C04	0.006	94	0.016	68	26269	0.44	22014	0.86	2506	62.0
SRP1848-C10	0.057	96	0.036	75	27465	0.91	15966	0.27	2326	5.6
SRP1848-B09	0.073	97	0.027	74	25152	0.46	25213	0.99	1424	78.0
SRP1848-C05	0.073	92	0.021	62	26836	0.52	15199	0.35	4134	4.8
SRP1848-F02	0.054	92	0.009	54	25714	0.62	14911	0.19	2741	2.6
SRP1848-F08	0.061	94	0.024	77	26483	0.91	21024	1.07	NB	NB
SRP1848-D07	0.075	94	0.032	71	25738	0.77	24272	0.92	NB	NB
SRP1848-F11	0.054	91	0.017	70	26774	0.75	21790	0.47	1762	4.6
SRP1848-F09	0.056	93	0.050	79	23816	0.36	24178	0.75	1671	90.7
SRP1848-D10	0.016	90	0.012	54	26468	0.48	20578	0.52	1859	13.0
SRP1848-G01	0.070	91	0.022	66	27406	0.98	20913	0.56	1993	4.6
SRP1848-B06	0.058	95	0.022	72	25070	0.67	26767	1.21	NB	NB
SRP1848-D03	0.160	98	0.038	76	25977	1.90	14130	0.58	3170	9.5
SRP1848-B07	0.079	96	0.038	73	25612	0.66	25491	1.05	NB	NB
SRP1848-E02	0.046	93	0.025	71	23847	0.53	18717	0.59	1473	21.0
SRP1848-B03	0.050	94	0.028	66	26338	0.82	17228	0.41	2722	6.4
SRP1848-E01	0.088	92	0.029	72	26430	1.01	22420	0.96	NB	NB
SRP1848-B05	0.065	94	0.040	72	24536	0.65	21871	0.64	NB	NB
SRP1848-D09	0.042	91	0.023	70	24966	0.46	21306	0.65	NB	NB
SRP1848-F06	0.066	94	0.032	77	25598	0.87	26528	0.86	NB	NB
SRP1848-G10	0.046	97	0.019	79	25269	0.49	14163	0.24	2891	4.3
SRP1848-G04	0.051	92	0.016	75	25156	0.76	12538	0.25	1999	2.5
SRP1848-G06	0.057	96	0.026	81	25838	0.63	12830	0.31	1857	11.1
SRP1848-G07	0.058	94	0.038	78	24939	0.78	13668	0.35	1978	2.9
SRP1848-G09	0.073	97	0.036	83	25066	0.59	17685	0.35	2184	6.4
SRP1848-G11	0.040	97	0.023	84	27191	0.68	11837	0.26	2744	7.6

TABLE 8
Affinity-matured antibodies from initial leads (Trastuzumab HC
framework): Kinetic binding results
	Biacore Kinetics
	Variant ID	ka (1/Ms)	kd (1/s)	KD (M)
	SRP1848-A01	8.29E+05	1.55E−03	1.87E−09
	SRP1848-A02	5.25E+05	8.82E−03	1.68E−08
	SRP1848-A07	1.01E+06	8.66E−04	8.55E−10
	SRP1848-C03	1.36E+06	1.52E−03	1.11E−09
	SRP1848-F04	8.15E+05	1.08E−03	1.32E−09
	SRP1848-B04	7.80E+05	1.17E−03	1.50E−09
	SRP1848-B11	1.22E+06	1.86E−03	1.52E−09
	SRP1848-F07	1.60E+06	2.49E−03	1.56E−09
	SRP1848-E06	9.44E+05	1.54E−03	1.63E−09
	SRP1848-A09	7.30E+05	1.33E−03	1.82E−09
	SRP1848-E07	1.25E+06	2.40E−03	1.91E−09
	SRP1848-G03	9.90E+05	1.97E−03	1.99E−09
	SRP1848-A04	1.61E+06	3.26E−03	2.03E−09
	SRP1848-H01	6.59E+05	1.39E−03	2.11E−09
	SRP1848-B10	6.81E+05	1.48E−03	2.18E−09
	SRP1848-C07	8.56E+05	1.89E−03	2.21E−09
	SRP1848-F05	6.56E+05	1.57E−03	2.40E−09
	SRP1848-D02	8.51E+05	2.05E−03	2.41E−09
	SRP1848-A08	4.93E+05	1.19E−03	2.42E−09
	SRP1848-E03	6.88E+05	1.83E−03	2.67E−09
	SRP1848-A10	1.20E+06	3.30E−03	2.74E−09
	SRP1848-F10	8.72E+05	2.47E−03	2.83E−09
	SRP1848-D05	6.75E+05	1.98E−03	2.93E−09
	SRP1848-C01	7.30E+05	2.23E−03	3.05E−09
	SRP1848-F01	1.14E+06	3.62E−03	3.18E−09
	SRP1848-D04	4.97E+05	1.73E−03	3.48E−09
	SRP1848-E05	7.16E+05	2.51E−03	3.51E−09
	SRP1848-A06	1.37E+06	4.83E−03	3.51E−09
	SRP1848-B01	1.13E+06	4.16E−03	3.67E−09
	SRP1848-C04	1.29E+06	4.99E−03	3.86E−09
	SRP1848-C10	8.99E+05	3.63E−03	4.03E−09
	SRP1848-B09	1.55E+06	6.61E−03	4.26E−09
	SRP1848-C05	1.06E+06	4.54E−03	4.29E−09
	SRP1848-F02	1.42E+06	6.37E−03	4.49E−09
	SRP1848-F08	5.94E+05	2.72E−03	4.58E−09
	SRP1848-D07	1.09E+06	5.11E−03	4.70E−09
	SRP1848-F11	8.28E+05	3.90E−03	4.71E−09
	SRP1848-F09	1.40E+06	6.79E−03	4.85E−09
	SRP1848-D10	1.13E+06	5.58E−03	4.95E−09
	SRP1848-G01	4.44E+05	2.26E−03	5.09E−09
	SRP1848-B06	6.20E+05	3.17E−03	5.10E−09
	SRP1848-D03	1.03E+06	5.35E−03	5.19E−09
	SRP1848-B07	7.06E+05	3.78E−03	5.35E−09
	SRP1848-E02	1.14E+06	7.07E−03	6.21E−09
	SRP1848-B03	1.13E+06	8.59E−03	7.63E−09
	SRP1848-E01	6.64E+05	5.22E−03	7.87E−09
	SRP1848-B05	9.76E+05	8.85E−03	9.07E−09
	SRP1848-D09	1.07E+06	1.08E−02	1.01E−08
	SRP1848-F06	4.56E+05	7.75E−03	1.70E−08
	SRP1848-G10	7.58E+05	3.45E−03	4.55E−09
	SRP1848-G04	5.91E+05	3.79E−03	6.40E−09
	SRP1848-G06	5.69E+05	3.81E−03	6.70E−09
	SRP1848-G07	6.05E+05	4.51E−03	7.45E−09
	SRP1848-G09	8.56E+05	6.46E−03	7.56E−09
	SRP1848-G11	6.96E+05	6.37E−03	9.14E−09

TABLE 9
Results obtained with humanized 6D1 (2060) antibodies.
		Igrov 2° Antibody
		Cell Killing,
	Biacore kinetics	Nb-SC239
		kd		EC50
SRP	ka (1/Ms)	(1/s)	KD (M)	(nM)	span (%)
SRP2060-E10	5.82E+05	1.20E−03	2.06E−09	0.061	68
SRP2060-E05	5.41E+05	1.58E−03	2.92E−09	0.22	71
SRP2060-B01	5.61E+05	1.47E−03	2.62E−09	0.045	76
SRP2060-A06	5.47E+−5	7.29E−03	1.33E−08	0.013	66

Example 8

Sequences

Table 10 provides sequences referred to herein.

TABLE 10
Sequences
SEQ
ID
NO:	Molecule	Region	Scheme	Sequence
1	Human folate			MAQRMTTQLLLLLVWVAVVGEAQTRIAW
	receptor alpha			ARTELLNVCMNAKHHKEKPGPEDKLHEQ
	(hFOLR1)			CRPWRKNACCSTNTSQEAHKDVSYLYRF
				NWNHCGEMAPACKRHFIQDTCLYECSPN
				LGPWIQQVDQSWRKERVLNVPLCKEDCE
				QWWEDCRTSYTCKSNWHKGWNWTSGFNK
				CAVGAACQPFHEYEPTPTVLCNEIWTHS
				YKVSNYSRGSGRCIQMWFDPAQGNPNEE
				VARFYAAAMSGAGPWAAWPFLLSLALML
				LWLLS
2	Cynomolgus			MAQRMTTQLLLLLVWVAVVGEAQTRTAR
	folate			ARTELLNVCMNAKHHKEKPGPEDKLHEQ
	receptor alpha			CRPWKKNACCSTNTSQEAHKDVSYLYRF
				NWNHCGEMAPACKRHFIQDTCLYECSPN
				LGPWIQQVDQSWRKERVLNVPLCKEDCE
				RWWEDCRTSYTCKSNWHKGWNWTSGFNK
				CPVGAACQPFHEYEPTPTVLCNEIWTYS
				YKVSNYSRGSGRCIQMWFDPAQGNPNEE
				VARFYAAAMSGAGPWAAWPLLLSLALTL
				LWLLS
3	Murine folate			MAHLMTVQLLLLVMWMAECAQSRATRAR
	receptor alpha			TELLNVCMDAKHHKEKPGPEDNLHDQCS
				PWKTNSCCSTNTSQEAHKDISYLYRFNW
				NHCGTMTSECKRHFIQDTCLYECSPNLG
				PWIQQVDQSWRKERILDVPLCKEDCQQW
				WEDCQSSFTCKSNWHKGWNWSSGHNECP
				VGASCHPFTFYFPTSAALCEEIWSHSYK
				LSNYSRGSGRCIQMWFDPAQGNPNEEVA
				RFYAEAMSGAGFHGTWPLLCSLSLVLLW
				VIS
4	SRP1848-A01	CDR-H1	Chothia	GFNITRY
5	SRP1848-A02	CDR-H1	Chothia	GFNISGF
6	SRP1848-A04	CDR-H1	Chothia	GFNIDQS
7	SRP1848-A06	CDR-H1	Chothia	GFNIGNS
8	SRP1848-A07	CDR-H1	Chothia	GFNIGYH
9	SRP1848-A08	CDR-H1	Chothia	GSNIRKH
10	SRP1848-A09	CDR-H1	Chothia	GENIRKQ
11	SRP1848-A10	CDR-H1	Chothia	GENIRKY
12	SRP1848-B01	CDR-H1	Chothia	GFNIRNY
13	SRP1848-B03	CDR-H1	Chothia	GFNISMK
14	SRP1848-B04	CDR-H1	Chothia	SFNISNH
15	SRP1848-B05	CDR-H1	Chothia	GFNISNY
16	SRP1848-B06	CDR-H1	Chothia	GFNISNY
17	SRP1848-B07	CDR-H1	Chothia	GFNISRF
18	SRP1848-B09	CDR-H1	Chothia	GFNITNY
19	SRP1848-B10	CDR-H1	Chothia	GFNTTTK
20	SRP1848-B11	CDR-H1	Chothia	GFNIGNN
21	SRP1848-001	CDR-H1	Chothia	GFNIGNS
22	SRP1848-0O3	CDR-H1	Chothia	GFNIGVY
23	SRP1848-004	CDR-H1	Chothia	GFNIRHY
24	SRP1848-005	CDR-H1	Chothia	GFNIRKY
25	SRP1848-007	CDR-H1	Chothia	GFNIRKY
26	SRP1848-C10	CDR-H1	Chothia	GFNIRTY
27	SRP1848-D02	CDR-H1	Chothia	GFNISHN
28	SRP1848-D03	CDR-H1	Chothia	GFNIRYF
29	SRP1848-D04	CDR-H1	Chothia	GFNISHY
30	SRP1848-D05	CDR-H1	Chothia	GFNISIS
31	SRP1848-D07	CDR-H1	Chothia	GFNISKY
32	SRP1848-D09	CDR-H1	Chothia	GFNISNY
33	SRP1848-D10	CDR-H1	Chothia	GFNISRN
34	SRP1848-E01	CDR-H1	Chothia	GFNITNK
35	SRP1848-E02	CDR-H1	Chothia	GFNIGKY
36	SRP1848-E03	CDR-H1	Chothia	GFNIGNY
37	SRP1848-E05	CDR-H1	Chothia	GFNIGVY
38	SRP1848-E06	CDR-H1	Chothia	GFNINRY
39	SRP1848-E07	CDR-H1	Chothia	GFNIRKS
40	SRP1848-F01	CDR-H1	Chothia	GFNIRTY
41	SRP1848-F02	CDR-H1	Chothia	GFNIRTY
42	SRP1848-F04	CDR-H1	Chothia	GFNISNY
43	SRP1848-F05	CDR-H1	Chothia	GFNISKS
44	SRP1848-F06	CDR-H1	Chothia	GFNISLS
45	SRP1848-F07	CDR-H1	Chothia	GFNISNH
46	SRP1848-F08	CDR-H1	Chothia	GFNISNH
47	SRP1848-F09	CDR-H1	Chothia	GFNISNH
48	SRP1848-F10	CDR-H1	Chothia	GFNISNN
49	SRP1848-F11	CDR-H1	Chothia	GFNISNN
50	SRP1848-G01	CDR-H1	Chothia	GFNISRH
51	SRP1848-G03	CDR-H1	Chothia	GFNISTY
52	SRP1848-G04	CDR-H1	Chothia	GFNIHST
53	SRP1848-G06	CDR-H1	Chothia	GFNIRST
54	SRP1848-G07	CDR-H1	Chothia	GFNIHST
55	SRP1848-G09	CDR-H1	Chothia	GFNIRGT
56	SRP1848-G10	CDR-H1	Chothia	GFNIRST
57	SRP1848-G11	CDR-H1	Chothia	GFNISST
58	SRP1848-H01	CDR-H1	Chothia	GFNIRTQ
59	SRP2060-E10	CDR-H1	Chothia	GFSLSTFGM
60	SRP2060-E05	CDR-H1	Chothia	GFSLSTFGM
61	SRP2060-B01	CDR-H1	Chothia	GFSLSTFGM
62	SRP2060-A06	CDR-H1	Chothia	GFSLSTFGM
63	SRP1848-A01	CDR-H1	Kabat	RYSIH
64	SRP1848-A02	CDR-H1	Kabat	GFRIH
65	SRP1848-A04	CDR-H1	Kabat	QSSIH
66	SRP1848-A06	CDR-H1	Kabat	NSYIH
67	SRP1848-A07	CDR-H1	Kabat	YHSIH
68	SRP1848-A08	CDR-H1	Kabat	KHSIH
69	SRP1848-A09	CDR-H1	Kabat	KQSIH
70	SRP1848-A10	CDR-H1	Kabat	KYSIH
71	SRP1848-B01	CDR-H1	Kabat	NYSIH
72	SRP1848-B03	CDR-H1	Kabat	MKYIH
73	SRP1848-B04	CDR-H1	Kabat	NHSIH
74	SRP1848-B05	CDR-H1	Kabat	NYYIH
75	SRP1848-B06	CDR-H1	Kabat	NYYIH
76	SRP1848-B07	CDR-H1	Kabat	RFYIH
77	SRP1848-B09	CDR-H1	Kabat	NYYIH
78	SRP1848-B10	CDR-H1	Kabat	TKSIH
79	SRP1848-B11	CDR-H1	Kabat	NNSIH
80	SRP1848-001	CDR-H1	Kabat	NSYIH
81	SRP1848-0O3	CDR-H1	Kabat	VYSIH
82	SRP1848-004	CDR-H1	Kabat	HYSIH
83	SRP1848-005	CDR-H1	Kabat	KYSIH
84	SRP1848-007	CDR-H1	Kabat	KYSIH
85	SRP1848-C10	CDR-H1	Kabat	TYYIH
86	SRP1848-D02	CDR-H1	Kabat	HNYIH
87	SRP1848-D03	CDR-H1	Kabat	YFSIH
88	SRP1848-D04	CDR-H1	Kabat	HYSIH
89	SRP1848-D05	CDR-H1	Kabat	ISYIH
90	SRP1848-D07	CDR-H1	Kabat	KYYIH
91	SRP1848-D09	CDR-H1	Kabat	NYYIH
92	SRP1848-D10	CDR-H1	Kabat	RNSIH
93	SRP1848-E01	CDR-H1	Kabat	NKYIH
94	SRP1848-E02	CDR-H1	Kabat	KYSIH
95	SRP1848-E03	CDR-H1	Kabat	NYYIH
96	SRP1848-E05	CDR-H1	Kabat	VYYIH
97	SRP1848-E06	CDR-H1	Kabat	RYYIH
98	SRP1848-E07	CDR-H1	Kabat	KSSIH
99	SRP1848-F01	CDR-H1	Kabat	TYSIH
100	SRP1848-F02	CDR-H1	Kabat	TYSIH
101	SRP1848-F04	CDR-H1	Kabat	NYSIH
102	SRP1848-F05	CDR-H1	Kabat	KSSIH
103	SRP1848-F06	CDR-H1	Kabat	LSYIH
104	SRP1848-F07	CDR-H1	Kabat	NHSIH
105	SRP1848-F08	CDR-H1	Kabat	NHSIH
106	SRP1848-F09	CDR-H1	Kabat	NHYIH
107	SRP1848-F10	CDR-H1	Kabat	NNSIH
108	SRP1848-F11	CDR-H1	Kabat	NNYIH
109	SRP1848-G01	CDR-H1	Kabat	RHSIH
110	SRP1848-G03	CDR-H1	Kabat	TYYIH
111	SRP1848-G04	CDR-H1	Kabat	STDIH
112	SRP1848-G06	CDR-H1	Kabat	STDIH
113	SRP1848-G07	CDR-H1	Kabat	STDIH
114	SRP1848-G09	CDR-H1	Kabat	GTDIH
115	SRP1848-G10	CDR-H1	Kabat	STDIH
116	SRP1848-G11	CDR-H1	Kabat	STDIH
117	SRP1848-H01	CDR-H1	Kabat	TQSIH
118	SRP2060-E10	CDR-H1	Kabat	TFGMGVG
119	SRP2060-E05	CDR-H1	Kabat	TFGMGVG
120	SRP2060-B01	CDR-H1	Kabat	TFGMGVG
121	SRP2060-A06	CDR-H1	Kabat	TFGMGVG
122	SRP1848-A01	CDR-H2	Chothia	LPESGG
123	SRP1848-A02	CDR-H2	Chothia	YPESGA
124	SRP1848-A04	CDR-H2	Chothia	YPVDGT
125	SRP1848-A06	CDR-H2	Chothia	TPIDGN
126	SRP1848-A07	CDR-H2	Chothia	FPVDGT
127	SRP1848-A08	CDR-H2	Chothia	YPNDGT
128	SRP1848-A09	CDR-H2	Chothia	FPNDGT
129	SRP1848-A10	CDR-H2	Chothia	FPIDDI
130	SRP1848-B01	CDR-H2	Chothia	YPVDGI
131	SRP1848-B03	CDR-H2	Chothia	TPIDGM
132	SRP1848-B04	CDR-H2	Chothia	YPVDGI
133	SRP1848-B05	CDR-H2	Chothia	SPIDGY
134	SRP1848-B06	CDR-H2	Chothia	TPIDGY
135	SRP1848-B07	CDR-H2	Chothia	SPYDGF
136	SRP1848-B09	CDR-H2	Chothia	TPVDGY
137	SRP1848-B10	CDR-H2	Chothia	YPRDGI
138	SRP1848-B11	CDR-H2	Chothia	SPIDGF
139	SRP1848-001	CDR-H2	Chothia	TPNDGY
140	SRP1848-0O3	CDR-H2	Chothia	YPIDGN
141	SRP1848-004	CDR-H2	Chothia	YPGPGN
142	SRP1848-005	CDR-H2	Chothia	FPIDGI
143	SRP1848-007	CDR-H2	Chothia	FPIDGI
144	SRP1848-C10	CDR-H2	Chothia	SPIDGY
145	SRP1848-D02	CDR-H2	Chothia	TPQDGY
146	SRP1848-D03	CDR-H2	Chothia	FPNDGS
147	SRP1848-D04	CDR-H2	Chothia	YPRDGI
148	SRP1848-D05	CDR-H2	Chothia	SPIDGY
149	SRP1848-D07	CDR-H2	Chothia	SPNDGY
150	SRP1848-D09	CDR-H2	Chothia	SPNDGY
151	SRP1848-D10	CDR-H2	Chothia	SPNDGT
152	SRP1848-E01	CDR-H2	Chothia	TPFDGF
153	SRP1848-E02	CDR-H2	Chothia	YPNDGN
154	SRP1848-E03	CDR-H2	Chothia	TPRDGF
155	SRP1848-E05	CDR-H2	Chothia	TPNDGY
156	SRP1848-E06	CDR-H2	Chothia	TPNDGY
157	SRP1848-E07	CDR-H2	Chothia	FPYDGS
158	SRP1848-F01	CDR-H2	Chothia	FPNDGT
159	SRP1848-F02	CDR-H2	Chothia	FPNDGT
160	SRP1848-F04	CDR-H2	Chothia	YPIDGI
161	SRP1848-F05	CDR-H2	Chothia	YPNDGS
162	SRP1848-F06	CDR-H2	Chothia	SPIDGN
163	SRP1848-F07	CDR-H2	Chothia	YPNDGI
164	SRP1848-F08	CDR-H2	Chothia	YPVDGI
165	SRP1848-F09	CDR-H2	Chothia	SPLDGY
166	SRP1848-F10	CDR-H2	Chothia	FPNDGY
167	SRP1848-F11	CDR-H2	Chothia	TPIDGN
168	SRP1848-G01	CDR-H2	Chothia	APNDGS
169	SRP1848-G03	CDR-H2	Chothia	TPSDGF
170	SRP1848-G04	CDR-H2	Chothia	TPAGGA
171	SRP1848-G06	CDR-H2	Chothia	TPAGGA
172	SRP1848-G07	CDR-H2	Chothia	TPAGGA
173	SRP1848-G09	CDR-H2	Chothia	TPAGGA
174	SRP1848-G10	CDR-H2	Chothia	TPAGGA
175	SRP1848-G11	CDR-H2	Chothia	TPAGGA
176	SRP1848-H01	CDR-H2	Chothia	FPIDGI
177	SRP2060-E10	CDR-H2	Chothia	WWDDD
178	SRP2060-E05	CDR-H2	Chothia	WWDDD
179	SRP2060-B01	CDR-H2	Chothia	WWDDD
180	SRP2060-A06	CDR-H2	Chothia	WWDDD
181	SRP1848-A01	CDR-H2	Kabat	GILPESGGTSYADSVKG
182	SRP1848-A02	CDR-H2	Kabat	GIYPESGATYYADSVKG
183	SRP1848-A04	CDR-H2	Kabat	VIYPVDGTTDYADSVKG
184	SRP1848-A06	CDR-H2	Kabat	GITPIDGNTDYADSVKG
185	SRP1848-A07	CDR-H2	Kabat	EIFPVDGTTDYADSVKG
186	SRP1848-A08	CDR-H2	Kabat	SIYPNDGTTDYADSVKG
187	SRP1848-A09	CDR-H2	Kabat	SIFPNDGTTDYADSVKG
188	SRP1848-A10	CDR-H2	Kabat	DIFPIDDITDYADSVKG
189	SRP1848-B01	CDR-H2	Kabat	EIYPVDGITDYADSVKG
190	SRP1848-B03	CDR-H2	Kabat	GITPIDGMTDYADSVKG
191	SRP1848-B04	CDR-H2	Kabat	EIYPVDGITDYADSVKG
192	SRP1848-B05	CDR-H2	Kabat	GISPIDGYTDYADSMKG
193	SRP1848-B06	CDR-H2	Kabat	GITPIDGYTDYADSVKG
194	SRP1848-B07	CDR-H2	Kabat	GISPYDGFTDYADSVKG
195	SRP1848-B09	CDR-H2	Kabat	GITPVDGYTDYADRVKG
196	SRP1848-B10	CDR-H2	Kabat	EIYPRDGITDYADSVKG
197	SRP1848-B11	CDR-H2	Kabat	DISPIDGFTDYADSVKG
198	SRP1848-001	CDR-H2	Kabat	GVTPNDGYTDYADSVKG
199	SRP1848-0O3	CDR-H2	Kabat	EIYPIDGNTDYADSVKG
200	SRP1848-004	CDR-H2	Kabat	ElYPGPGNTDYADSVKG
201	SRP1848-005	CDR-H2	Kabat	DIFPIDGINDYADSVKG
202	SRP1848-007	CDR-H2	Kabat	DIFPIDGITDYADSVKG
203	SRP1848-C10	CDR-H2	Kabat	GISPIDGYTDYADSMKG
204	SRP1848-D02	CDR-H2	Kabat	GITPQDGYTDYADSVKG
205	SRP1848-D03	CDR-H2	Kabat	DIFPNDGSTDYADSVKG
206	SRP1848-D04	CDR-H2	Kabat	EIYPRDGITDYADSVKG
207	SRP1848-D05	CDR-H2	Kabat	GISPIDGYTDYADSVKG
208	SRP1848-D07	CDR-H2	Kabat	GISPNDGYTDYADSVKG
209	SRP1848-D09	CDR-H2	Kabat	GISPNDGYTDYADSVKG
210	SRP1848-D10	CDR-H2	Kabat	WISPNDGTTDYADSVKG
211	SRP1848-E01	CDR-H2	Kabat	GITPFDGFTDYADSVKG
212	SRP1848-E02	CDR-H2	Kabat	EIYPNDGNTDYADSVKG
213	SRP1848-E03	CDR-H2	Kabat	GITPRDGFTDYADSVKG
214	SRP1848-E05	CDR-H2	Kabat	GITPNDGYTDYADSVKG
215	SRP1848-E06	CDR-H2	Kabat	GITPNDGYTDYADSVEG
216	SRP1848-E07	CDR-H2	Kabat	EIFPYDGSTDYADNVKG
217	SRP1848-F01	CDR-H2	Kabat	SIFPNDGTTDYADSVKG
218	SRP1848-F02	CDR-H2	Kabat	SIFPNDGTTDYADSVKG
219	SRP1848-F04	CDR-H2	Kabat	EIYPIDGITDYADSVKG
220	SRP1848-F05	CDR-H2	Kabat	EIYPNDGSTDYADSVKG
221	SRP1848-F06	CDR-H2	Kabat	GISPIDGNTDYADSVKG
222	SRP1848-F07	CDR-H2	Kabat	EIYPNDGITDYADSVKG
223	SRP1848-F08	CDR-H2	Kabat	EIYPVDGITDYADSVKG
224	SRP1848-F09	CDR-H2	Kabat	GISPLDGYTDYADSVKG
225	SRP1848-F10	CDR-H2	Kabat	SIFPNDGYTDYADSVKG
226	SRP1848-F11	CDR-H2	Kabat	GITPIDGNTDYADSVKG
227	SRP1848-G01	CDR-H2	Kabat	WIAPNDGSTDYADSVKG
228	SRP1848-G03	CDR-H2	Kabat	GITPSDGFTDYADSVKG
229	SRP1848-G04	CDR-H2	Kabat	YITPAGGATFYADSVKG
230	SRP1848-G06	CDR-H2	Kabat	YITPAGGATYYADNVKG
231	SRP1848-G07	CDR-H2	Kabat	YITPAGGATWYADSVKG
232	SRP1848-G09	CDR-H2	Kabat	YITPAGGATFYADSVKG
233	SRP1848-G10	CDR-H2	Kabat	YITPAGGATYYADSVKG
234	SRP1848-G11	CDR-H2	Kabat	YITPAGGATWYADSVKG
235	SRP1848-H01	CDR-H2	Kabat	DIFPIDGITDYADSVKG
236	SRP2060-E10	CDR-H2	Kabat	HIWWDDDKYYHPALKG
237	SRP2060-E05	CDR-H2	Kabat	HIWWDDDKYYHPALKG
238	SRP2060-B01	CDR-H2	Kabat	HIWWDDDKYYHPALKG
239	SRP2060-A06	CDR-H2	Kabat	HIWWDDDKYYYPALKG
240	SRP1848-A01	CDR-H3		HIYPWDWFSNYVLDY
241	SRP1848-A02	CDR-H3		HLYVWDWVLDHVLDY
242	SRP1848-A04	CDR-H3		GAWSWRSGYGYYIDY
243	SRP1848-A06	CDR-H3		GAWSWRSGYGYYIDY
244	SRP1848-A07	CDR-H3		GFWAWRSGYGYYLDY
245	SRP1848-A08	CDR-H3		GSWFWRAGYGYYLDY
246	SRP1848-A09	CDR-H3		GSWFWRSGYGYFLEY
247	SRP1848-A10	CDR-H3		GSWSWPSGHSYYLDY
248	SRP1848-B01	CDR-H3		GFWSWPSGYSYFLDY
249	SRP1848-B03	CDR-H3		GSWSWPSGYSYYLDY
250	SRP1848-B04	CDR-H3		GRYSWRAGYSYYLDY
251	SRP1848-B05	CDR-H3		GSWFWQSGYGYYLDY
252	SRP1848-B06	CDR-H3		GFWSWPSGYGYYQDY
253	SRP1848-B07	CDR-H3		GSWSWPAGYGYYQDY
254	SRP1848-B09	CDR-H3		GAWSWRSGYGYYMDY
255	SRP1848-B10	CDR-H3		GGWHWRSGYSYYLDY
256	SRP1848-B11	CDR-H3		GSWSWRAGYGYYLDY
257	SRP1848-C01	CDR-H3		GSWFWRAGYGYYLDY
258	SRP1848-C03	CDR-H3		GSWAWRSGYSYYLDY
259	SRP1848-C04	CDR-H3		GSLSWRAGYGYYLDY
260	SRP1848-C05	CDR-H3		GSWSWKAGYGYYLDY
261	SRP1848-C07	CDR-H3		GSWSWPAGYGYYQDY
262	SRP1848-C10	CDR-H3		GSWSWPAGYGYYLDY
263	SRP1848-D02	CDR-H3		GAWSWRAGYGYYLDY
264	SRP1848-D03	CDR-H3		GHWSWPSGYWYYLDY
265	SRP1848-D04	CDR-H3		GYWFWRSGYGYYLDY
266	SRP1848-D05	CDR-H3		GSWSWRAGYGYYLDY
267	SRP1848-D07	CDR-H3		GFWAWRSGYGYYLDY
268	SRP1848-D09	CDR-H3		GSWSWRHGYGYYLDY
269	SRP1848-D10	CDR-H3		GAWSWRSGYGYYIDY
270	SRP1848-E01	CDR-H3		GSWSWPAGYGYYQDY
271	SRP1848-E02	CDR-H3		GSWSWRSGYGYYLDY
272	SRP1848-E03	CDR-H3		GSWSWPAGHSYYLDY
273	SRP1848-E05	CDR-H3		GFWAWRSGYGYYLDY
274	SRP1848-E06	CDR-H3		GTWSWPSGHSYYLDY
275	SRP1848-E07	CDR-H3		GAWSWRSGYGYYIDY
276	SRP1848-F01	CDR-H3		GSWAWRAGYSYYLDY
277	SRP1848-F02	CDR-H3		GSWSWQAGYGYYLDY
278	SRP1848-F04	CDR-H3		GSWFWRSGYGYYLDY
279	SRP1848-F05	CDR-H3		GSWAWRSGYSYFLDY
280	SRP1848-F06	CDR-H3		GFWAWRSGYGYYLDY
281	SRP1848-F07	CDR-H3		GSWDWRSGYSYYLDY
282	SRP1848-F08	CDR-H3		GSWYWQSGYSYYLDY
283	SRP1848-F09	CDR-H3		GAWSWRSGYGYYIDY
284	SRP1848-F10	CDR-H3		GSWFWRSGYGYYLDY
285	SRP1848-F11	CDR-H3		GSWYWRAGYGYYLDY
286	SRP1848-G01	CDR-H3		GSWAWRSGYSYFLDY
287	SRP1848-G03	CDR-H3		GSWSWPSGHGYFLDY
288	SRP1848-G04	CDR-H3		YPYWFAGYMDY
289	SRP1848-G06	CDR-H3		QPYWFAGYMDY
290	SRP1848-G07	CDR-H3		YPFWFAGYMDY
291	SRP1848-G09	CDR-H3		HEYWFSGYMDY
292	SRP1848-G10	CDR-H3		YPYWFAGYIDY
293	SRP1848-G11	CDR-H3		YPYWFSGYMDY
294	SRP1848-H01	CDR-H3		GSWSWPSGMDYYLDY
295	SRP2060-E10	CDR-H3		NHFPHYYGSSHWYFNV
296	SRP2060-E05	CDR-H3		NHFPHYYGSSHWYFNV
297	SRP2060-B01	CDR-H3		NHFPHYYGSSHWYFNV
298	SRP2060-A06	CDR-H3		NHFPHYYGSSHWYFDV
299	trastuzumab	CDR-LI		RASQDVNTAVA
300	H6D1-LC4	CDR-L1		KASQDINSYLS
301	H6D1-LC5	CDR-L1		KASQDINSYLS
302	trastuzumab	CDR-L2		SASFLYS
303	H6D1-LC4	CDR-L3		RANRLVD
304	H6D1-LC5	CDR-L2		RANRLVD
305	trastuzumab	CDR-L3		QQHYTTPPT
306	H6D1-LC4	CDR-L3		LQYDEFPYT
307	H6D1-LC5	CDR-L3		LQYDEFPYT
308	SRP1848-A01	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ITRYSIHWVRQAPGKGLEWVAGILPESG
				GTSYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARHIYPWDWFSNYVLD
				YWGQGTLVTVSS
309	SRP1848-A02	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISGFRIHWVRQAPGKGLEWVAGIYPESG
				ATYYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARHLYVWDWVLDHVLD
				YWGQGTLVTVSS
310	SRP1848-A04	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISSIDQHWVRQAPGKGLEWVGVIYPVDG
				TTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGAWSWRSGYGYYID
				YWGQGTLVTVSS
311	SRP1848-A06	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IGNSYIHWVRQAPGKGLEWVGGITPIDG
				NTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGAWSWRSGYGYYID
				YWGQGTLVTVSS
312	SRP1848-A07	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IGYHSIHWVRQAPGKGLEWVGEIFPVDG
				TTDYADSVKGRFTISADTSKNTAYLHMN
				SLRAEDTAVYYCARGFWAWRSGYGYYLD
				YWGQGTLVTVSS
313	SRP1848-A08	VH		EVQLVESGGGLVQPGGSLRLSCAASGSN
				IRKHSIHWVRQAPGKGLEWVGSIYPNDG
				TTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWFWRAGYGYYLD
				YWGQGTLVTVSS
314	SRP1848-A09	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRKQSIHWVRQAPGKGLEWVGSIFPNDG
				TTDYADSVKGRFTISADTSKNTAYLQVN
				SLRAEDTAVYYCARGSWFWRSGYGYFLE
				YWGQGTLVTVSS
315	SRP1848-A10	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRKYSIHWARQAPGKGLEWVGDIFPIDD
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPSGHSYYLD
				YWGQGTLVTVSS
316	SRP1848-B01	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRNYSIHWVRQAPGKGLEWVGEIYPVDG
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGFWSWPSGYSYFLD
				YWGQGTLVTVSS
317	SRP1848-B03	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISMKYIHWVRQAPGKGLEWVGGITPIDG
				MTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPSGYSYYLD
				YWGQGTLVTVSS
318	SRP1848-B04	VH		EVQLVESGGGLVQPGGSLRLSCAASSFN
				ISNHSIHWVRQAPGKGLEWVGEIYPVDG
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGRYSWRAGYSYYLD
				YWGQGTLVTVSS
319	SRP1848-B05	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISNYYIHWVRQAPGKGLEWVGGISPIDG
				YTDYADSMKGRFISADTSKNTAYLQMS
				SLRAEDTAVYYCARGSWFWQSGYGYYLD
				YWGQGTLVTVSS
320	SRP1848-B06	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISNYYIHWVRQAPGKGLEWVGGITPIDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGFWSWPSGYGYYQD
				YWGQGTLVTVSS
321	SRP1848-B07	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISRFYIHWVRQAPGKGLEWVGGISPYDG
				FTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPAGYGYYQD
				YWGQGTLVTVSS
322	SRP1848-B09	VH		EVQLVESGGGLVQPGGSLRLSCAAGGFN
				ITNYYIHWVRQAPGKGLEWVGGITPVDG
				YTDYADRVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGAWSWRSGYGYYMD
				YWGQGTLVTVSS
323	SRP1848-B10	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				TTTKSIHWVRQAPGKGLEWVGEIYPRDG
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGGWHWRSGYSYYLD
				YWGQGTLVTVSS
324	SRP1848-B11	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IGNNSIHWVRQAPGKGLEWVGDISPIDG
				FTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWRAGYGYYLD
				YWGQGTLVTVSS
325	SRP1848-C01	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IGNSYIHWVRQAPGKGLEWVGGVTPNDG
				YTDYADSVKGRFTISADTSKNTTYLQMN
				SLRAEDTAVYYCARGSWFWRAGYGYYLD
				YWGQGALVTVSS
326	SRP1848-C03	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IGVYSIHWVRQAPGKGLEWVGEIYPIDG
				NTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWAWRSGYSYYLD
				YWGQGTLVTVSS
327	SRP1848-C04	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRHYSIHWVRQAPGKGLEWVGEIYPGPG
				NTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSLSWRAGYGYYLD
				YWGQGTLVTVSS
328	SRP1848-C05	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRKYSIHWVRQAPGKGLEWVGDIFPIDG
				INDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWKAGYGYYLD
				YWGQGTLVTVSS
329	SRP1848-C07	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRKYSIHWVRQAPGKGLEWVGDIFPIDG
				ITDYADSMKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPAGYGYYQD
				YWGQGTLVTVSS
330	SRP1848-C10	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRTYYIHWVRQAPGKGLEWVGGISPIDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPAGYGYYLD
				YWGQGTLVTVSS
331	SRP1848-D02	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISHNYIHWVRQAPGKGLEWVGGITPQDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				RLRAEDTAVYYCARGAWSWRAGYGYYLD
				YWGQGTLVTVSS
332	SRP1848-D03	VH		EVQLVESGGGVVQPGGSLRLSCAASGFN
				IRYFSIHWVRQAPGKGLEWVGDIFPNDG
				STDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEETAVYYCARGHWSWPSGYWYYLD
				YWGQGTLVTVSS
333	SRP1848-D04	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISHYSIHWVRQAPGKGLEWVGEIYPRDG
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLSAEDTAVYYCARGYWFWRSGYGYYLD
				YWGQGTLVTVSS
334	SRP1848-D05	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISISYIHWVRQAPGKGLEWVGGISPIDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWRAGYGYYLD
				YWGQGTLVTVSS
335	SRP1848-D07	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISKYYIHWVRQAPGKGLEWVGGISPNDG
				YTDYADSVKGRFAISADTSKNTAYLQMN
				SLRAEDTAVYYCARGFWAWRSGYGYYLD
				YWGQGTLVTVSS
336	SRP1848-D09	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISNYYIHWVRQAPGKGLEWVGGISPNDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWRHGYGYYLD
				YWGQGTLVTVSS
337	SRP1848-D10	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISRNSIHWVRQAPGKGLEWVGWISPNDG
				TTDYADSVKGRFTISADGSKNTAYLQMN
				SLRAEDTAVYYCARGAWSWRSGYGYYID
				YWGQGTLVTVSS
338	SRP1848-E01	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ITNKYIHWVRQAPGKGLEWVGGITPFDG
				FTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPAGYGYYQD
				YWGQGTLVTVSS
339	SRP1848-E02	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IGKYSIHWVRQAPGKGLEWVGEIYPNDG
				NTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWRSGYGYYLD
				YWGQGTLVTVSS
340	SRP1848-E03	VH		EVQLVESGGGLAQPGGSLRLSCAASGFN
				IGNYYIHWVRQAPGKGLEWVGGITPRDG
				FTDYADSVKGRFTISADTSKNTAYLQVN
				SLRAEDTAVYYCARGSWSWPAGHSYYLD
				YWGQGTLVTVSS
341	SRP1848-E05	VH		EVQLVESGGGLVQPGGSLRVSCAASGFN
				IGVYYIHWVRQAPGKGLEWVGGITPNDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGFWAWRSGYGYYLD
				YWGQGTLVTVSS
342	SRP1848-E06	VH		EVQLVESGGGLVQPSGSLRLSCAASGFN
				INRYYIHWVRQAPGKGLEWVGGITPNDG
				YTDYADSVEGRFTTSADTSKNTAYLQMN
				SLRAEDTAVYYCARGTWSWPSGHSYYLD
				YWGQGTLVTVSS
343	SRP1848-E07	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRKSSIHWVRQAPGKGLEWVGEIFPYDG
				STDYADNVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGAWSWRSGYGYYID
				YWGQGTLVTVSS
344	SRP1848-F01	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRTYSIHWVRQAPGKGLEWVGSIFPNDG
				TTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWAWRAGYSYYLD
				YWGQGTLVTVSS
345	SRP1848-F02	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRTYSIHWVRQAPGKGLEWVGSIFPNDG
				TTDYADSVKGRLTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWQAGYGYYLD
				YWGQGTLVTVSS
346	SRP1848-F04	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISNYSIHWVRQAPGKGLEWVGEIYPIDG
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWFWRSGYGYYLD
				YWGQGTLVTVSS
347	SRP1848-F05	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISKSSIHWVRQAPGKGLEWVGEIYPNDG
				STDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWAWRSGYSYFLD
				YWGQGTLVTVSS
348	SRP1848-F06	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISLSYIHWVRQAPGKGLEWVGGISPIDG
				NTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGFWAWRSGYGYYLD
				YWGQGTLVTVSS
349	SRP1848-F07	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISNHSIHWVRQAPGKGLEWVGEIYPNDG
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLSAEDTAVYYCARGSWDWRSGYSYYLD
				YWGQGTLVTVSS
350	SRP1848-F08	VH		EVQLVESGGGLVQPGGSLRLSCAAGGFN
				ISNHSIHWVRQAPGKGVEWVGEIYPVDG
				ITDYADSVKGRFTISADTSKNTAYLRMN
				SLRAEDTAVYYCARGSWYWQSGYSYYLD
				YWGQGTLVTVSS
351	SRP1848-F09	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISNHYIHWVRQAPGKGLEWVGGISPLDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGAWSWRSGYGYYID
				YWGQGTLVTVSS
352	SRP1848-F10	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISNNSIHWVRQAPGKGLEWVGSIFPNDG
				YTDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWFWRSGYGYYLD
				YWGQGTLVTVSS
353	SRP1848-F11	VH		ISNNYIHWVRQAPGKGLEWVGGITPIDG
				NTDYADSVKGRFTISADTSMNTAYLQMN
				SLRAEDTAVYYCARGSWYWRAGYGYYLD
				YWGQGALVTVSS
354	SRP1848-G01	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISRHSIHWVRQAPGKGLEWVGWIAPNDG
				STDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWAWRSGYSYFLD
				YWGQGTLVTVSS
355	SRP1848-G03	VH		ISTYYIHWVRQAPGKGLEWVGGITPSDG
				FTDYADSVKGRSTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPSGHGYFLD
				YWGQGTLVTVSS
356	SRP1848-G04	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IHSTDIHWVRQAPGKGLEWVAYITPAGG
				ATFYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARYPYWFAGYMDYWGQ
				GTLVTVSS
357	SRP1848-G06	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRSTDIHWVRQAPGKGLEWVAYITPAGG
				ATYYADNVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARQPYWFAGYMDYWGQ
				GTLVTVSS
358	SRP1848-G07	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IHSTDIHWVRQAPGKGLEWVAYITPAGG
				ATWYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARYPFWFAGYMDYWGQ
				GTLVTVSS
359	SRP1848-G09	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRGTDIHWVRQAPGKGLEWVAYITPAGG
				ATFYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARHEYWFSGYMDYWGQ
				GTLVTVSS
360	SRP1848-G10	VH		EVQLVESGGGLVQPGSSLRLSCAASGFN
				IRSTDIHWVRQAPGKGLEWVAYITPAGG
				ATYYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARYPYWFAGYIDYWGQ
				GTLVTVSS
361	SRP1848-G11	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				ISSTDIHWVRQAPGKGLEWVAYITPAGG
				ATWYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARYPYWFSGYMDYWGQ
				GTLVTVSS
362	SRP1848-H01	VH		EVQLVESGGGLVQPGGSLRLSCAASGFN
				IRTQSIHWVRQAPGKGLEWIGDIFPIDG
				ITDYADSVKGRFTISADTSKNTAYLQMN
				SLRAEDTAVYYCARGSWSWPSGMDYYLD
				YWGQGTLVTVSS
363	SRP2060-E10	VH		EVQLLESGGGLVQPGGSLRLSCAFSGFS
				LSTFGMGVGWVRQAPGKGLEWVSHIWWD
				DDKYYHPALKGRFTISKDNSKNTVYLQM
				NSLRAEDTAVYYCGRNHFPHYYGSSHWY
				FNVWGQGTTVTVSS
364	SRP2060-E05	VH		EVQLLESGGGLVQPGGSLRLSCAFSGFS
				LSTFGMGVGWVRQAPGKGLEWVSHIWWD
				DDKYYHPALKGRFTVSKDNSKNTVYLQM
				NSLRAEDTAVYYCGRNHFPHYYGSSHWY
				FNVWGQGTTVTVSS
365	SRP2060-B01	VH		EVQLLESGGGLVQPGGSLRLSCALSGFS
				LSTFGMGVGWVRQATGKGLEWVSHIWWD
				DDKYYHPALKGRFTISKDNSKNTVHLQM
				NSLRAEDTAVYYCGRNHFPHYYGSSHWY
				FNVWGQGTTVTVSS
366	SRP2060-A06	VH		EVQLLESGGGLVQPGGSLRLSCAFSGFS
				LSTFGMGVGWVRQAPGKGLEWVGHIWWD
				DDKYYYPALKGRFTISKDNSKNTVYLQM
				NSLRAEDTAVYYCGRNHFPHYYGSSHWY
				FDVWGQGTTVTVSS
367	trastuzumab	VL		DIQMTQSPSSLSASVGDRVTITCRASQD
				VNTAVAWYQQKPGKAPKLLIYSASFLYS
				GVPSRFSGSRSGTDFTLTISSLQPEDFA
				TYYCQQHYTTPPTFGQGTKVEIK
368	H6D1-LC4	VL		EIVMTQSPATLSLSPGERATLSCKASQD
				INSYLSWYQQKPGQAPRLLIYRANRLVD
				GIPARFSGSGSGTDYTLTISSLEPEDFA
				VYYCLQYDEFPYTFGGGTKVEIK
369	H6D1-LC5	VL		DIQMTQSPSTLSASVGDRVTITCKASQD
				INSYLSWYQQKPGKAPKLLIYRANRLVD
				GVPSRFSGSGSGTEFTLTISSLQPDDFA
				TYYCLQYDEFPYTFGGGTKVEIK
370	Human IgG1 HC			ASTKGPSVFPLAPSSKSTSGGTAALGCL
	Constant			VKDYFPEPVTVSWNSGALTSGVHTFPAV
				LQSSGLYSLSSVVTVPSSSLGTQTYICN
				VNHKPSNTKVDKKVEPKSCDKTHTCPPC
				PAPELLGGPSVFLEPPKPKDTLMISRTP
				EVTCVVVDVSHEDPEVKFNWYVDGVEVH
				NAKTKPREEQYNSTYRVVSVLTVLHQDW
				LNGKEYKCKVSNKALPAPIEKTISKAKG
				QPREPQVYTLPPSREEMTKNQVSLTCLV
				KGFYPSDIAVEWESNGQPENNYKTTPPV
				LDSDGSFFLYSKLTVDKSRWQQGNVFSC
				SVMHEALHNHYTQKSLSLSPGK
371	Human IgG LC			RTVAAPSVFIEPPSDEQLKSGTASVVCL
	Constant Ckappa			LNNFYPREAKVQWKVDNALQSGNSQESV
				TEQDSKDSTYSLSSTLTLSKADYEKHKV
				YACEVTHQGLSSPVTKSFNRGEC
372	Mouse IgG1 HC			AKTTPPSVYPLAPGSAAQTNSMVTLGCL
	Constant			VKGYFPEPVTVTWNSGSLSSGVHTFPAV
				LQSDLYTLSSSVTVPSSTWPSETVTCNV
				AHPASTKVDKKIVPRDCGCKPCICTVP
				EVSSVFIFPPKPKDVLTITLTPKVTCVV
				VDISKDDPEVQFSWFVDDVEVHTAQTQP
				REEQFNSTFRSVSELPIMHQDWLNGKEF
				KCRVNSAAFPAPIEKTISKTKGRPKAPQ
				VYTIPPPKEQMAKDKVSLTCMITDFFPE
				DITVEWQWNGQPAENYKNTQPIMDTDGS
				YFVYSKLNVQKSNWEAGNTFTCSVLHEG
				LHNHHTEKSLSHSPG
373	Mouse IgG LC			RADAAPTVSIEPPSSEQLTSGGASVVCF
	Constant Ckappa			LNNFYPKD NVKWKIDGSERQNGVLNSW
				TDQDSKDSTYSMSSTLTLTKDEYERHNS
				YTCEATHKTSTSPIVKSFNRNEC
374	Kappa LC			HMTVAAPSVFIFPPSDEQLKSGTASVVC
				LLNNFYPREAKVQWKVDNALQSGNSQES
				VTEQDSKDSTYSLSSTLTLSKADYEKHK
				VYACEVTHQGLSSPVTKSFNRGEC
375	Lambda LD			GQPKAAPSVTLFPPSSEELQANKATLVC
				LISDFYPGAVTVAWKADSSPVKAGVETT
				TPSKQSNNKYAASSYLSLTPEQWKSHRS
				YSCQVTHEGSTVEKTVAPTECS
376	FlagHis Tag			GSGDYKDDDDKGSGHHHHHH
377	Linker			GGGGSGGGGSGGGGS
378	Linker			AAGSDQEPKSS
379	1848-B10-VH-	scFv		MEVQLVESGGGLVQPGGSLRLSCAASGF
	(G4S)3-VL			NTTTKSIHWVRQAPGKGLEWVGEIYPRD
				GITDYADSVKGRFTISADTSKNTAYLQM
				NSLRAEDTAVYYCARGGWHWRSGYSYYL
				DYWGQGTLVTVSSGGGGSGGGGSGGGGS
				DIQMTQSPSSLSASVGDRVTITCRASQD
				VNTAVAWYQQKPGKAPKLLIYSASFLYS
				GVPSRFSGSRSGTDFTLTISSLQPEDFA
				TYYCQQHYTTPPTFGQGTKVEIK
380	1848-B10-VL-	scFv		MDIQMTQSPSSLSASVGDRVTITCRASQ
	(G4S)3-VH			DVNTAVAWYQQKPGKAPKLLIYSASFLY
				SGVPSRFSGSRSGTDFTLTISSLQPEDF
				ATYYCQQHYTTPPTFGQGTKVEIKGGGG
				SGGGGSGGGGSEVQLVESGGGLVQPGGS
				LRLSCAASGFNTTTKSIHWVRQAPGKGL
				EWVGEIYPRDGITDYADSVKGRFTISAD
				TSKNTAYLQMNSLRAEDTAVYYCARGGW
				HWRSGYSYYLDYWGQGTLVTVSS
381	1848-B10-VH-	scFv-Fc		MEVQLVESGGGLVQPGGSLRLSCAASGF
	(G4S)3-VL			NTTTKSIHWVRQAPGKGLEWVGEIYPRD
				GITDYADSVKGRFTISADTSKNTAYLQM
				NSLRAEDTAVYYCARGGWHWRSGYSYYL
				DYWGQGTLVTVSSGGGGSGGGGSGGGGS
				DIQMTQSPSSLSASVGDRVTITCRASQD
				VNTAVAWYQQKPGKAPKLLIYSASFLYS
				GVPSRFSGSRSGTDFTLTISSLQPEDFA
				TYYCQQHYTTPPTFGQGTKVEIKAAGSD
				QEPKSSDKTHTCPPCPAPELLGGPSVFL
				FPPKPKDTLMISRTPEVTCVVVDVSHED
				PEVKFNWYVDGVEVHNAKTKPREEQYNS
				TYRVVSVLTVLHQDWLNGKEYKCKVSNK
				ALPAPIEKTISKAKGQPREPQVYTLPPS
				REEMTKNQVSLTCLVKGFYPSDIAVEWE
				SNGQPENNYKTTPPVLDSDGSFFLYSKL
				TVDKSRWQQGNVFSCSVMHEALHNHYTQ
				KSLSLSPGK
382	1848-B10-VL-	scFv-Fc		MDIQMTQSPSSLSASVGDRVTITCRASQ
	(G4S)3-VH			DVNTAVAWYQQKPGKAPKLLIYSASFLY
				SGVPSRFSGSRSGTDFTLTISSLQPEDF
				ATYYCQQHYTTPPTFGQGTKVEIKGGGG
				SGGGGSGGGGSEVQLVESGGGLVQPGGS
				LRLSCAASGFNTTTKSIHWVRQAPGKGL
				EWVGEIYPRDGITDYADSVKGRFTISAD
				TSKNTAYLQMNSLRAEDTAVYYCARGGW
				HWRSGYSYYLDYWGQGTLVTVSSAAGSD
				QEPKSSDKTHTCPPCPAPELLGGPSVFL
				FPPKPKDTLMISRTPEVTCVVVDVSHED
				PEVKFNWYVDGVEVHNAKTKPREEQYNS
				TYRVVSVLTVLHQDWLNGKEYKCKVSNK
				ALPAPIEKTISKAKGQPREPQVYTLPPS
				REEMTKNQVSLTCLVKGFYPSDIAVEWE
				SNGQPENNYKTTPPVLDSDGSFFLYSKL
				TVDKSRWQQGNVFSCSVMHEALHNHYTQ
				KSLSLSPGK

EQUIVALENTS

The disclosure set forth above may encompass multiple distinct inventions with independent utility. Although each of these inventions has been disclosed in its preferred form(s), the specific embodiments thereof as disclosed and illustrated herein are not to be considered in a limiting sense, because numerous variations are possible. The subject matter of the inventions includes all novel and nonobvious combinations and subcombinations of the various elements, features, functions, and/or properties disclosed herein. The following claims particularly point out certain combinations and subcombinations regarded as novel and nonobvious. Inventions embodied in other combinations and subcombinations of features, functions, elements, and/or properties may be claimed in this application, in applications claiming priority from this application, or in related applications. Such claims, whether directed to a different invention or to the same invention, and whether broader, narrower, equal, or different in scope in comparison to the original claims, also are regarded as included within the subject matter of the inventions of the present disclosure.

One or more features from any embodiments described herein or in the figures may be combined with one or more features of any other embodiments described herein or in the figures without departing from the scope of the invention.

All publications, patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Claims

1. An isolated antibody that specifically binds to folate receptor alpha (FOLR1), wherein the antibody comprises a CDR-H3 sequence selected from: a sequence defined by the consensus sequence G-α2-α3-α4-W-α6-α7-G-α9-α10-Y-α12-α13-α14-Y, where α2 is G, S, A, F, H, R, T, or Y; α3 is W, L, or Y; α4 is S, A, F, Y, H, or D; α6 is R, P, Q, or K; α7 is S, A, or H; α9 is Y, H, or M; α10 is G, S, D, or W; α12 is Y or F; α13 is L, I, Q, or M; and α14 is D or E;a sequence defined by the consensus sequence G-α2-α3-α4-W-α6-α7-G-α9-α10-Y-α12-α13-α14-Y, where α2 is G or S; α3 is W; α4 is S or H; α6 is R or P; α7 is S; α9 is Y or M; α10 is G, S, or D; α12 is Y; α13 is L; and α14 is D; anda sequence selected from SEQ ID NOs: 240-298, or a variant thereof having three, two, or one amino acid substitution(s).

2. The antibody of claim 1, wherein the CDR-H3 sequence is a sequence selected from SEQ ID NOs: 240-298.

3. The antibody of claim 1, wherein the antibody comprises a Chothia CDR-H2 sequence selected from: a sequence defined by the consensus sequence ε1-ε2-ε3-ε4-ε5-ε6, where ε1 is Y, T, F, S, or A; ε2 is P; ε3 is N, I, V, R, Y, F, G, L, Q, or S; ε4 is D or P; ε5 is G or D; and ε6 is Y, I, T, N, F, S, or M;a sequence defined by the consensus sequence ε1-ε2-ε3-ε4-ε5-ε6, where ε1 is Y or F; ε2 is P; ε3 is N, I, or R; ε4 is D; ε5 is G; and ε6 is Y or I; anda sequence selected from SEQ ID NOs: 122-180, or a variant thereof having two or one amino acid substitutions(s).

4. The antibody of claim 3, wherein the Chothia CDR-H2 sequence is a sequence selected from SEQ ID NOs: 122-180.

5. The antibody of claim 1, wherein the antibody comprises a Chothia CDR-H1 sequence selected from: a sequence defined by the consensus sequence γ1-γ2-γ3-γ4-γ5-γ6-γ7, where γ1 is G or S; γ2 is F or S; γ3 is N; γ4 is I or T; γ5 is S, R, G, T, N, or D; γ6 is N, K, T, R, H, Y, L, M, Q, or V; and γ7 is Y, H, S, N, K, F, or Q;a sequence defined by the consensus sequence γ1-γ2-γ3-γ4-γ5-γ6-γ7, where γ1 is G; γ2 is F; γ3 is N; γ4 is I or T; γ5 is S, R, or T; γ6 is N or T; and γ7 is Y, K, or Q; anda sequence selected from SEQ ID NOs: 4-62, or a variant thereof having two or one amino acid substitutions(s).

6. The antibody of claim 5, wherein the Chothia CDR-H1 sequence is a sequence selected from SEQ ID NOs: 4-62.

7. The antibody of claim 1, wherein the antibody comprises a Kabat CDR-H2 sequence selected from: a sequence defined by the consensus sequence θ1-θ2-θ3-θ4-θ5-θ6-θ7-θ8-θ9-D-Y-A-D-θ14-θ15-θ16-G, where θ1 is G, E, D, W, S, or V; θ2 is I or V; θ3 is Y, T, F, S, or A; θ4 is P; θ5 is N, I, V, R, Y, F, G, L, Q, or S; θ6 is D or P; θ7 is G or D; θ8 is Y, I, T, N, F, S, or M; θ9 is T or N; θ14 is S, R, or N; θ15 is V or M; and θ16 is K or E;a sequence defined by the consensus sequence θ1-θ2-θ3-θ4-θ5-θ6-θ7-θ8-θ9-D-Y-A-D-θ14-θ15-θ16-G, where θ1 is G, E, or D; θ2 is I; θ3 is Y or F; θ4 is P; θ5 is N, I, or R; θ6 is D; θ7 is G; θ8 is Y or I; θ9 is T; θ14 is S; θ15 is V; and θ16 is K; anda sequence selected from SEQ ID NOs: 181-239, or a variant thereof having three, two, or one amino acid substitutions(s).

8. The antibody of claim 7, wherein the Kabat CDR-H2 sequence is a sequence selected from SEQ ID NOs: 181-239.

9. The antibody of claim 1, wherein the antibody comprises a Kabat CDR-H1 sequence selected from: a sequence defined by the consensus sequence ζ1-ζ2-ζ3-ζ4-ζ5, where ζ1 is N, K, T, R, H, Y, L, M, Q, or V; ζ2 is Y, H, S, N, K, F, or Q; ζ3 is S or Y; ζ4 is I; and ζ5 is H;a sequence defined by the consensus sequence ζ1-ζ2-ζ3-ζ4-ζ5, where ζ1 is N or T; ζ2 is Y, K, or Q; ζ3 is S; ζ4 is I; and ζ5 is H; anda sequence selected from SEQ ID NOs: 63-121, or a variant thereof having two or one amino acid substitutions.

10. The antibody of claim 9, wherein the Kabat CDR-H1 sequence is a sequence selected from SEQ ID NOs: 63-121.

11. The antibody of claim 1, wherein the antibody comprises a CDR-L3 sequence selected from SEQ ID NOs: 305-307, or a variant thereof having three, two, or one amino acid substitution(s).

12. The antibody of claim 1, wherein the antibody comprises a CDR-L2 sequence selected from a sequence selected from SEQ ID NOs: 302-304, or a variant thereof having two or one amino acid substitution(s).

13. The antibody of claim 1, wherein the antibody comprises a CDR-L1 sequence selected from a sequence selected from SEQ ID NOs: 299-301, or a variant thereof having three, two, or one amino acid substitution(s).

14. The antibody of claim 1, wherein the antibody comprises: a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 4 and 63; a CDR-H2 comprising one or more of SEQ ID NOs: 122 and 181; and a CDR-H3 comprising SEQ ID NO: 240;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 5 and 64; a CDR-H2 comprising one or more of SEQ ID NOs: 123 and 182; and a CDR-H3 comprising SEQ ID NO: 241;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 6 and 65; a CDR-H2 comprising one or more of SEQ ID NOs: 124 and 183; and a CDR-H3 comprising SEQ ID NO: 242;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 7 and 66; a CDR-H2 comprising one or more of SEQ ID NOs: 125 and 184; and a CDR-H3 comprising SEQ ID NO: 243;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 8 and 67; a CDR-H2 comprising one or more of SEQ ID NOs: 126 and 185; and a CDR-H3 comprising SEQ ID NO: 244;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 9 and 68; a CDR-H2 comprising one or more of SEQ ID NOs: 127 and 186; and a CDR-H3 comprising SEQ ID NO: 245;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 10 and 69; a CDR-H2 comprising one or more of SEQ ID NOs: 128 and 187; and a CDR-H3 comprising SEQ ID NO: 246;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 11 and 70; a CDR-H2 comprising one or more of SEQ ID NOs: 129 and 188; and a CDR-H3 comprising SEQ ID NO: 247;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 12 and 71; a CDR-H2 comprising one or more of SEQ ID NOs: 130 and 189; and a CDR-H3 comprising SEQ ID NO: 248;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 13 and 72; a CDR-H2 comprising one or more of SEQ ID NOs: 131 and 190; and a CDR-H3 comprising SEQ ID NO: 249;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 14 and 73; a CDR-H2 comprising one or more of SEQ ID NOs: 132 and 191; and a CDR-H3 comprising SEQ ID NO: 250;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 15 and 74; a CDR-H2 comprising one or more of SEQ ID NOs: 133 and 192; and a CDR-H3 comprising SEQ ID NO: 251;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 16 and 75; a CDR-H2 comprising one or more of SEQ ID NOs: 134 and 193; and a CDR-H3 comprising SEQ ID NO: 252;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 17 and 76; a CDR-H2 comprising one or more of SEQ ID NOs: 135 and 194; and a CDR-H3 comprising SEQ ID NO: 253;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 18 and 77; a CDR-H2 comprising one or more of SEQ ID NOs: 136 and 195; and a CDR-H3 comprising SEQ ID NO: 254;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 19 and 78; a CDR-H2 comprising one or more of SEQ ID NOs: 137 and 196; and a CDR-H3 comprising SEQ ID NO: 255;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 20 and 79; a CDR-H2 comprising one or more of SEQ ID NOs: 138 and 197; and a CDR-H3 comprising SEQ ID NO: 256;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 21 and 80; a CDR-H2 comprising one or more of SEQ ID NOs: 139 and 198; and a CDR-H3 comprising SEQ ID NO: 257;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 22 and 81; a CDR-H2 comprising one or more of SEQ ID NOs: 140 and 199; and a CDR-H3 comprising SEQ ID NO: 258;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 23 and 82; a CDR-H2 comprising one or more of SEQ ID NOs: 141 and 200; and a CDR-H3 comprising SEQ ID NO: 259;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 24 and 83; a CDR-H2 comprising one or more of SEQ ID NOs: 142 and 201; and a CDR-H3 comprising SEQ ID NO: 260;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 25 and 84; a CDR-H2 comprising one or more of SEQ ID NOs: 143 and 202; and a CDR-H3 comprising SEQ ID NO: 261;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 26 and 85; a CDR-H2 comprising one or more of SEQ ID NOs: 144 and 203; and a CDR-H3 comprising SEQ ID NO: 262;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 27 and 86; a CDR-H2 comprising one or more of SEQ ID NOs: 145 and 204; and a CDR-H3 comprising SEQ ID NO: 263;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 28 and 87; a CDR-H2 comprising one or more of SEQ ID NOs: 146 and 205; and a CDR-H3 comprising SEQ ID NO: 264;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 29 and 88; a CDR-H2 comprising one or more of SEQ ID NOs: 147 and 206; and a CDR-H3 comprising SEQ ID NO: 265;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 30 and 89; a CDR-H2 comprising one or more of SEQ ID NOs: 148 and 207; and a CDR-H3 comprising SEQ ID NO: 266;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 31 and 90; a CDR-H2 comprising one or more of SEQ ID NOs: 149 and 208; and a CDR-H3 comprising SEQ ID NO: 267;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 32 and 91; a CDR-H2 comprising one or more of SEQ ID NOs: 150 and 209; and a CDR-H3 comprising SEQ ID NO: 268;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 33 and 92; a CDR-H2 comprising one or more of SEQ ID NOs: 151 and 210; and a CDR-H3 comprising SEQ ID NO: 269;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 34 and 93; a CDR-H2 comprising one or more of SEQ ID NOs: 152 and 211; and a CDR-H3 comprising SEQ ID NO: 270;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 35 and 94; a CDR-H2 comprising one or more of SEQ ID NOs: 153 and 212; and a CDR-H3 comprising SEQ ID NO: 271;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 36 and 95; a CDR-H2 comprising one or more of SEQ ID NOs: 154 and 213; and a CDR-H3 comprising SEQ ID NO: 272;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 37 and 96; a CDR-H2 comprising one or more of SEQ ID NOs: 155 and 214; and a CDR-H3 comprising SEQ ID NO: 273;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 38 and 97; a CDR-H2 comprising one or more of SEQ ID NOs: 156 and 215; and a CDR-H3 comprising SEQ ID NO: 274;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 39 and 98; a CDR-H2 comprising one or more of SEQ ID NOs: 157 and 216; and a CDR-H3 comprising SEQ ID NO: 275;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 40 and 99; a CDR-H2 comprising one or more of SEQ ID NOs: 158 and 217; and a CDR-H3 comprising SEQ ID NO: 276;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 41 and 100; a CDR-H2 comprising one or more of SEQ ID NOs: 159 and 218; and a CDR-H3 comprising SEQ ID NO: 277;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 42 and 101; a CDR-H2 comprising one or more of SEQ ID NOs: 160 and 219; and a CDR-H3 comprising SEQ ID NO: 278;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 43 and 102; a CDR-H2 comprising one or more of SEQ ID NOs: 161 and 220; and a CDR-H3 comprising SEQ ID NO: 279;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 44 and 103; a CDR-H2 comprising one or more of SEQ ID NOs: 162 and 221; and a CDR-H3 comprising SEQ ID NO: 280;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 45 and 104; a CDR-H2 comprising one or more of SEQ ID NOs: 163 and 222; and a CDR-H3 comprising SEQ ID NO: 281;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 46 and 105; a CDR-H2 comprising one or more of SEQ ID NOs: 164 and 223; and a CDR-H3 comprising SEQ ID NO: 282;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 47 and 106; a CDR-H2 comprising one or more of SEQ ID NOs: 165 and 224; and a CDR-H3 comprising SEQ ID NO: 283;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 48 and 107; a CDR-H2 comprising one or more of SEQ ID NOs: 166 and 225; and a CDR-H3 comprising SEQ ID NO: 284;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 49 and 108; a CDR-H2 comprising one or more of SEQ ID NOs: 167 and 226; and a CDR-H3 comprising SEQ ID NO: 285;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 50 and 109; a CDR-H2 comprising one or more of SEQ ID NOs: 168 and 227; and a CDR-H3 comprising SEQ ID NO: 286;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 51 and 110; a CDR-H2 comprising one or more of SEQ ID NOs: 169 and 228; and a CDR-H3 comprising SEQ ID NO: 287;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 52 and 111; a CDR-H2 comprising one or more of SEQ ID NOs: 170 and 229; and a CDR-H3 comprising SEQ ID NO: 288;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 53 and 112; a CDR-H2 comprising one or more of SEQ ID NOs: 171 and 230; and a CDR-H3 comprising SEQ ID NO: 289;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 54 and 113; a CDR-H2 comprising one or more of SEQ ID NOs: 172 and 231; and a CDR-H3 comprising SEQ ID NO: 290;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 55 and 114; a CDR-H2 comprising one or more of SEQ ID NOs: 173 and 232; and a CDR-H3 comprising SEQ ID NO: 291;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 56 and 115; a CDR-H2 comprising one or more of SEQ ID NOs: 174 and 233; and a CDR-H3 comprising SEQ ID NO: 292;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 57 and 116; a CDR-H2 comprising one or more of SEQ ID NOs: 175 and 234; and a CDR-H3 comprising SEQ ID NO: 293;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 58 and 117; a CDR-H2 comprising one or more of SEQ ID NOs: 176 and 235; and a CDR-H3 comprising SEQ ID NO: 294;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 59 and 118; a CDR-H2 comprising one or more of SEQ ID NOs: 177 and 236; and a CDR-H3 comprising SEQ ID NO: 295;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 60 and 119; a CDR-H2 comprising one or more of SEQ ID NOs: 178 and 237; and a CDR-H3 comprising SEQ ID NO: 296;a VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 61 and 120; a CDR-H2 comprising one or more of SEQ ID NOs: 179 and 238; and a CDR-H3 comprising SEQ ID NO: 297; ora VH comprising: a CDR-H1 comprising one or more of SEQ ID NOs: 62 and 121; a CDR-H2 comprising one or more of SEQ ID NOs: 180 and 239; and a CDR-H3 comprising SEQ ID NO: 298.

15. The antibody of claim 14, wherein the VH is selected from SEQ ID NOs: 308-366, or a variant thereof having 20 or fewer amino acid substitutions.

16. The antibody of claim 14, wherein the antibody comprises: a VL comprising: a CDR-L1 comprising SEQ ID NO: 300; a CDR-L2 comprising SEQ ID NO: 303; and a CDR-L3 comprising SEQ ID NO: 306; ora VL comprising: a CDR-L1 comprising SEQ ID NO: 301; a CDR-L2 comprising SEQ ID NO: 304; and a CDR-L3 comprising SEQ ID NO: 307.

17. The antibody of claim 16 wherein the VL sequence is selected from SEQ ID NOs: 368 and 369, or a variant thereof having 20 or fewer amino acid substitutions.

18. An antibody comprising a VH region selected from SEQ ID NOs: 308-366, or a variant thereof having 20 or fewer amino acid substitutions, and a VL region selected from SEQ ID NOs: 367-369, or a variant thereof having 20 or fewer amino acid substitutions.

19. The antibody of claim 18, wherein: the VH region is SEQ ID NO: 308, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 309, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 310, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 311, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 312, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 313, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 314, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 315, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 316, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 317, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 318, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 319, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 320, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 321, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 322, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 323, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 324, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 325, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 326, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof; orthe VH region is SEQ ID NO: 327, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 328, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 329, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 330, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 331, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 332, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 333, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 334, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 335, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 336, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 337, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 338, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 339, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 340, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 341, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 342, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 343, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 344, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 345, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 346, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 347, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 348, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 349, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 350, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 351, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 352, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 353, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 354, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 355, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 356, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 357, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 358, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 359, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 360, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 361, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 362, or the variant thereof, and the VL region is SEQ ID NO: 367, or the variant thereof;the VH region is SEQ ID NO: 363, or the variant thereof, and the VL region is SEQ ID NO: 368, or the variant thereof;the VH region is SEQ ID NO: 364, or the variant thereof, and the VL region is SEQ ID NO: 368, or the variant thereof;the VH region is SEQ ID NO: 365, or the variant thereof, and the VL region is SEQ ID NO: 369, or the variant thereof; orthe VH region is SEQ ID NO: 366, or the variant thereof, and the VL region is SEQ ID NO: 369, or the variant thereof.

20. The antibody of claim 1, wherein the amino acid substitution is a conservative amino acid substitution.

21. The antibody of claim 1, wherein the antibody comprises at least one constant region domain.

22. The antibody of claim 21, wherein the constant region comprises a sequence selected from SEQ ID NOs: 370, 371, and 372.

23. The antibody of claim 1, wherein the antibody is a monoclonal antibody.

24. The antibody of claim 1, wherein the antibody is an IgA, an IgD, an IgE, an IgG, or an IgM.

25. The antibody of claim 1, wherein the antibody is humanized or human.

26. The antibody of claim 1, wherein the antibody is aglycosylated.

27. The antibody of claim 1, wherein the antibody is an antibody fragment.

28. The antibody of claim 27, wherein the antibody fragment is selected from an Fv fragment, a Fab fragment, a F(ab′)2 fragment, a Fab′ fragment, an scFv (sFv) fragment, and an scFv-Fc fragment.

29. The antibody of claim 28, wherein the antibody is an scFv fragment.

30. The antibody of claim 29, wherein the scFv fragment comprises a sequence selected from SEQ ID NO: 379 and SEQ ID NO: 380.

31. The antibody of claim 28, wherein the antibody is an scFv-Fc fragment.

32. The antibody of claim 31, wherein the scFv-Fc fragment comprises a sequence selected from SEQ ID NO: 381 and SEQ ID NO: 382.

33. The antibody of claim 1, wherein the antibody has a ka of about 2.90×105 M−1×sec−1 to about 9.64×109 M−1×sec−1 when associating with human folate receptor at a temperature of 25° C.

34. The antibody of claim 1, wherein the antibody has a kd of about 2.28×10−4 sec−1 to about 4.82×101 sec−1 when dissociating from human folate receptor at a temperature of 25° C.

35. The antibody of claim 1, wherein the antibody has a KD of about 2.26×10−11 M to about 7.20×10−9 M when bound to human folate receptor at a temperature of 25° C.

36. The antibody of claim 1, wherein the antibody specifically binds cynomolgus folate receptor.

37. The antibody of claim 40, wherein the antibody has a KD of about 0.19×10−9M to about 2.84×10−9 M when bound to cynomolgus folate receptor at a temperature of 25° C.

38. The antibody of claim 1, wherein the antibody specifically binds mouse folate receptor.

39. The antibody of claim 42, wherein the antibody has a KD of about 0.5×10−9 M to about 9.07×10−8 M when bound to mouse folate receptor at a temperature of 25° C.

40. A kit comprising an antibody of claim 1, and instructions for use of the antibody.

41. The kit of claim 40, wherein the antibody is lyophilized.

42. The kit of claim 41, further comprising a fluid for reconstitution of the lyophilized antibody.

43. A polynucleotide encoding an antibody of claim 1.

44. A vector comprising the polynucleotide of claim 43.

45. A recombinant host cell comprising the vector of claim 44.

46. The host cell of claim 45, wherein the host cell is selected from a bacterial cell, a fungal cell, and a mammalian cell.

47. The host cell of claim 46, wherein the host cell is selected from an E. coli cell, a Saccharomyces cerevisiae cell, and a CHO cell.

48. A cell-free expression reaction comprising the vector of claim 44.

49. A pharmaceutical composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.

50. A method of treating or preventing a disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of an antibody of claim 1.

51. A method of diagnosing a disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of an antibody of claim 1.

52. The method of claim 50, wherein the disease or condition is a cancer.

53. The method of claim 50, wherein the disease or condition is breast cancer.

54. The method of claim 50, wherein the disease or condition is triple-negative breast cancer (TNBC).

55. The method of claim 50, wherein the disease or condition is ovarian cancer.

56. The method of claim 50, wherein the disease or condition is lung cancer.

57. The method of claim 50, wherein the disease or condition is non-small cell lung cancer (NSCLC).

58. The method of claim 50, wherein the disease or condition is endometrial cancer.

59. A method of treating or preventing a disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition of claim 49.

60. A method of diagnosing a disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition of claim 49.