Antigen-binding molecules targeting thymic stromal lymphopoietin (TSLP)

Abstract

In various embodiments, the present disclosure provides polypeptides (e.g., antibodies and antigen-binding fragments thereof) that specifically bind to thymic stromal lymphopoietin (TSLP) (e.g., full-length human TSLP). In various embodiments, the present disclosure also provides fusion proteins comprising one or more of the polypeptides, polynucleotides encoding the polypeptides, vectors and host cells suitable for expressing the polypeptides, and methods of treating TSLP-associated diseases or conditions.

Description

[Technical Field]

[0001] Related Applications This application is a continuation of U.S. Provisional Patent Application No. 63 / 369,152, filed July 22, 2022; U.S. Provisional Patent Application No. 63 / 369,158, filed July 22, 2022; U.S. Provisional Patent Application No. 63 / 369,165, filed July 22, 2022; U.S. Provisional Patent Application No. 63 / 369,175, filed July 22, 2022; This application claims the benefit of U.S. Provisional Patent Application No. 63 / 369,202, filed July 22, 2022; U.S. Provisional Patent Application No. 63 / 369,203, filed July 22, 2022; U.S. Provisional Patent Application No. 63 / 369,204, filed July 22, 2022; and U.S. Provisional Patent Application No. 63 / 369,206, filed July 22, 2022.

[0002] This application claims priority to, and is a continuation-in-part of, U.S. Patent Application No. 18 / 356,891, filed July 21, 2023.

[0003] The entire teachings of the above applications are incorporated herein by reference. Without limiting the foregoing, this application incorporates by reference the sequence listings and code in Appendix A and B, respectively, of the above applications. For example, files claim.txt and fit_model.txt of U.S. Patent Application No. 18 / 356,891.

[0004] Incorporating XML material by reference This application incorporates by reference a sequence listing contained in the following extensible markup language (XML) file, filed concurrently herewith: a) File name: 57081061002_Sequence_Listing.xml; created on July 21, 2023, size 212,394 bytes.

[0005] Incorporation by reference of material in ASCII file format This application incorporates by reference the computer program listing contained in the following ASCII file, filed concurrently herewith: a) Filename: claim.py; uploaded July 22, 2023, available by selecting "claim.py" at https: / / zenodo.org / record / 8174726 / files / claim.py?download=1 or https: / / doi.org / 10.5281 / zenodo.8174726, size 4,774 bytes. The entire teachings of the above file are incorporated herein by reference in their entirety. b) Filename: fit_model.etab; uploaded July 22, 2023, available at https: / / zenodo.org / record / 8174726 / files / fit_model.etab?download=1 or by selecting "fit_model.etab" at https: / / doi.org / 10.5281 / zenodo.8174726, size 2,888,059 bytes. The entire teachings of the above file are incorporated herein by reference in their entirety. The entire teachings of the above files are incorporated herein by reference in their entirety. The script claim.py uses data from fit_model.etab and is therefore intended to be placed in the same folder as fit_model.etab and run in a Python environment. [Background technology]

[0006] Thymic stromal lymphopoietin (TSLP) is a cytokine implicated in the pathogenesis of various inflammatory conditions. TSLP is released upon epithelial exposure to insults such as allergens, initiating the downstream inflammatory cascade. The TSLP receptor (TSLPR) is expressed on a wide range of cell types, including airway smooth muscle cells (ASMCs), basophils, dendritic cells, eosinophils, type 2 innate lymphoid cells (ILC2s), hematopoietic progenitor cells, lymphocytes, macrophages, mast cells, and monocytes (Gauvreau et al., Thymic stromal lymphopoietin: its role and potential as a therapeutic target in asthma, Expert Opin Ther Targets 24(8):777-92(2020)). There is strong evidence that dysregulation of TSLP expression can lead to allergic diseases through the generation of type 2 inflammatory responses (Ziegler et al., The biology of thymic stromal lymphopoietin (TSLP), Adv Pharmacol. 66:129-55 (2013)). [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Gauvreau et al.,Thymic stromal lymphopoietin:its role and potential as a therapeutic target in asthma,Expert Opin Ther Targets 24(8):777-92(2020) [Non-patent document 2] Ziegler et al., The biology of thymic stromal lymphopoietin (TSLP), Adv Pharmacol.66:129-55(2013) Summary of the Invention [Problem to be solved by the invention]

[0008] There is a critical need for therapeutic agents to modulate (e.g., decrease or neutralize) thymic stromal lymphopoietin (TSLP) binding and / or activity. The present disclosure provides such agents. [Means for solving the problem]

[0009] The disclosure provided herein is based, in part, on the discovery that the polypeptides disclosed herein specifically bind to TSLP and demonstrate a variety of additional beneficial properties. Accordingly, the present disclosure generally relates to polypeptides (e.g., antibodies), compositions (e.g., pharmaceutical compositions), and methods useful for modulating (e.g., reducing or neutralizing) TSLP function and / or activity (e.g., in vivo).

[0010] Provided herein, among other things, are polypeptides (e.g., antibodies and antigen-binding fragments thereof) that specifically bind to TSLP (e.g., full-length human TSLP) or variants thereof. In some embodiments, the polypeptides disclosed herein bind to TSLP at the AB loop region and the C-terminal region of helix D.

[0011] In some embodiments, the polypeptide modulates (e.g., reduces, e.g., inhibits) binding between TSLP and TSLPR (e.g., alone or in a heterodimeric complex with interleukin-7 receptor alpha (IL-7Rα)). In some embodiments, the polypeptide modulates (e.g., reduces, e.g., inhibits) TSLP-mediated signaling in vitro and / or in vivo.

[0012] The present disclosure provides, inter alia, a polypeptide (e.g., an antibody or antigen-binding fragment thereof) that specifically binds to TSLP, K below 10 pM D binding affinity to TSLP characterized by (e.g., as measured by KinExA); Binding specificity to the AB loop region and the C-terminal region of helix D of TSLP; Neutralizing activity against TSLP (e.g., full-length human TSLP); or Inhibitory activity against TSLP-mediated signal transduction, or a combination of the above The present invention provides polypeptides having one or more properties selected from the following:

[0013] The present disclosure also provides, inter alia: SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of the above an immunoglobulin heavy chain variable domain (V) selected from H ) / immunoglobulin light chain variable domain (V L a polypeptide (e.g., an antibody or antigen-binding fragment thereof) comprising a paratope substantially similar to the paratope of an antibody comprising a combination of Also provided is a polypeptide that does not include a heavy chain comprising SEQ ID NO:81 and a light chain comprising SEQ ID NO:82.

[0014] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), or A combination of the above V to be selected from H / V L and a paratope substantially similar to the paratope of an antibody comprising a combination of wherein the polypeptide does not include a heavy chain comprising SEQ ID NO:81 and a light chain comprising SEQ ID NO:82.

[0015] The present disclosure also provides, inter alia: A V having HCDR1, HCDR2, and HCDR3 substantially similar to heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2), and heavy chain complementarity determining region 3 (HCDR3), respectively, of any one of the amino acid sequences of SEQ ID NOs: 4 to 17. H the amino acid sequence; and V comprising LCDR1, LCDR2 and LCDR3 substantially similar to the light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2) and light chain complementarity determining region 3 (LCDR3) of any one of the amino acid sequences of SEQ ID NOs: 20 to 30, respectively. L Amino acid sequence a polypeptide (e.g., an antibody or antigen-binding fragment thereof) comprising: Also provided is a polypeptide that does not include a heavy chain comprising SEQ ID NO:81 and a light chain comprising SEQ ID NO:82.

[0016] In some embodiments, the polypeptide is V comprising HCDR1, HCDR2 and HCDR3 substantially similar to HCDR1, HCDR2 and HCDR3, respectively, of any one of the amino acid sequences of SEQ ID NOs: 4 to 7. H the amino acid sequence; and V comprising LCDR1, LCDR2 and LCDR3 substantially similar to LCDR1, LCDR2 and LCDR3 of any one of the amino acid sequences of SEQ ID NOs: 20 to 22, respectively. L Amino acid sequence Includes.

[0017] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14) V to be selected from H / V L and HCDR1, HCDR2 and HCDR3, and LCDR1, LCDR2 and LCDR3 of antibodies comprising a combination of:

[0018] In some embodiments, the polypeptide further comprises: SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14) V to be selected from H / V L and a paratope having 100% sequence identity with the paratope of an antibody comprising a combination of:

[0019] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4) V to be selected from H / V L and HCDR1, HCDR2 and HCDR3, and LCDR1, LCDR2 and LCDR3 of antibodies comprising a combination of:

[0020] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4) V to be selected from H / V L and a paratope having 100% sequence identity with the paratope of an antibody comprising a combination of:

[0021] The present disclosure also provides, inter alia, SEQ ID NO: 2, wherein: X1 is not I; X2 is not W; X3 is not D; X4 is not S; X5 is not N; X6 is not K; X7 is not A; X8 is not A; X9 is not L; X 10 is not V; X 11 is not H, X 12 is not A; X 13 is not F; X 14 is not I, or any combination of the foregoing) H Also provided is a polypeptide (e.g., an antibody or antigen-binding fragment thereof) comprising:

[0022] In some embodiments, the polypeptide has the sequence of SEQ ID NO: 18, X 15 is not N; X 16 is not L; X 17 is not S; X 18 is not K; X 19 is not S; X 20 is not V; X 21 is not W; X 22 is not D; X 23 is not S; X 24 is not S; X 25 is not S; X 26 is not D; X 27 is not H, or any combination of the foregoing) L Includes.

[0023] The present disclosure also provides, inter alia, a V H V having at least 70% sequence identity with sequence SEQ ID NO: 19 LAlso provided are polypeptides (e.g., antibodies or antigen-binding fragments thereof) comprising the V H The sequence does not include SEQ ID NO: 3 or V L The sequence does not include SEQ ID NO: 19, or both.

[0024] The present disclosure also provides, inter alia: a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31, HCDR2 comprising the amino acid sequence of SEQ ID NO: 35, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 50; and b) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60, LCDR2 comprising the amino acid sequence of SEQ ID NO: 67, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0025] The present disclosure also provides, inter alia: a) HCDR1 consisting of the amino acid sequence of SEQ ID NO: 31, HCDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and HCDR3 consisting of the amino acid sequence of SEQ ID NO: 50; and b) LCDR1 consisting of the amino acid sequence of SEQ ID NO: 60, LCDR2 consisting of the amino acid sequence of SEQ ID NO: 67, and LCDR3 consisting of the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0026] In some embodiments, the polypeptide is a) V that is humanized, contains human framework regions, or a combination thereof H ; b) V that is humanized, contains human framework regions, or a combination thereof L , or Both a) and b) Includes.

[0027] In some embodiments, the polypeptide is a) V comprising the amino acid sequence of SEQ ID NO: 5 H ; b) V comprising the amino acid sequence of SEQ ID NO: 21 L , or Both a) and b) Includes.

[0028] In some embodiments, the antigen-binding fragment is a single-chain fragment variable (scFv), a variable heavy chain domain of the heavy chain (V HH ), fragment antigen binding (Fab), Fab' or F(ab')2.

[0029] In some embodiments, the polypeptide is a) an antibody heavy chain constant domain; b) an antibody light chain constant domain; Or both a) and b) Includes.

[0030] In some embodiments, the antibody heavy chain constant domain is an IgG1, IgG2, IgG3, or IgG4 constant domain.

[0031] In some embodiments, the antibody heavy chain constant domain is an IgG1 or IgG2 constant domain.

[0032] In some embodiments, the antibody heavy chain constant domain comprises one or more mutations that increase the serum half-life of the antibody or antigen-binding fragment thereof in humans. In some embodiments, the antibody heavy chain constant domain comprises amino acid substitutions at amino acid residues 252, 254, and 256 (wherein amino acid residues are numbered according to the EU index as in Kabat) with tyrosine, threonine, and glutamic acid, respectively, compared to a wild-type human IgG constant domain.

[0033] In some embodiments, the polypeptide is a) an antibody heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 92; b) an antibody light chain (LC) comprising the amino acid sequence of SEQ ID NO: 120; or Both a) and b) Includes.

[0034] In some embodiments, the polypeptide is a) an antibody heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 106; b) an antibody light chain (LC) comprising the amino acid sequence of SEQ ID NO: 120; or Both a) and b) Includes.

[0035] In some embodiments, the polypeptide specifically binds to TSLP.

[0036] In some embodiments, the polypeptide is a fusion protein.

[0037] In some embodiments, the present disclosure provides polynucleotides encoding the polypeptides disclosed herein, vectors comprising such polynucleotides, and / or host cells comprising such polynucleotides and / or vectors.

[0038] In some embodiments, the present disclosure provides a composition (e.g., a pharmaceutical composition) comprising a polypeptide disclosed herein. In some embodiments, the composition further comprises one or more pharmaceutical excipients, diluents, or carriers. In some embodiments, the composition further comprises one or more additional therapeutic agents (e.g., anti-inflammatory agents). In some embodiments, the polypeptide is conjugated (directly or via a linker) to one or more additional therapeutic agents (e.g., anti-inflammatory agents). In some embodiments, the one or more additional therapeutic agents comprise a corticosteroid, a beta-agonist, a muscarinic antagonist, an anti-inflammatory agent, an IL-4 and / or IL-13 antagonist (e.g., an antibody or antigen-binding fragment thereof that targets IL-13, IL-4, or IL-4R), or a combination thereof.

[0039] The present disclosure also provides, inter alia, methods of treating a subject in need of treatment (e.g., a subject having a TSLP-associated disease or condition), comprising administering to the subject an effective amount of a polypeptide disclosed herein and / or a composition (e.g., a pharmaceutical composition) comprising one or more polypeptides disclosed herein.

[0040] The present disclosure also provides, inter alia, methods for modulating (e.g., reducing) TSLP-mediated signaling in a cell (e.g., a cell of a subject), comprising contacting the cell with a polypeptide disclosed herein and / or a composition (e.g., a pharmaceutical composition) comprising one or more polypeptides disclosed herein.

[0041] In some embodiments, the subject has asthma, atopic dermatitis (AD), allergic conjunctivitis, chronic obstructive pulmonary disease (COPD), chronic spontaneous urticaria (CSU), rheumatoid arthritis (RA), rhinosinusitis (RS), eosinophilic esophagitis (EE), or food hypersensitivity. In some embodiments, the subject has moderate asthma. In some embodiments, the subject has severe asthma. In some embodiments, the subject has COPD. In some embodiments, the subject has COPD and / or either moderate asthma or severe asthma.

[0042] The present disclosure also provides, inter alia, a method for reducing binding of TSLP to the TSLP receptor (TSLPR) on a cell of a subject, comprising contacting the cell with an effective amount of a composition (e.g., a pharmaceutical composition) comprising one or more polypeptides disclosed herein.

[0043] The present disclosure also provides, inter alia, a method of making a polypeptide disclosed herein, comprising culturing a host cell containing a nucleotide sequence encoding the polypeptide under conditions such that the polypeptide is expressed in the host cell.

[0044] The present disclosure also provides, inter alia, a computer-implemented method that includes scoring a polypeptide comprising an amino acid sequence with a computationally binding optimized (CBO) model. The CBO model calculates a plurality of energy scores for each amino acid position of the amino acid sequence of the polypeptide, where the plurality of energy scores is based on substituting each of a plurality of different amino acids for an amino acid at a given position in the amino acid sequence. The CBO model further adds each calculated energy score to an array. The CBO model further generates a normalized sum of the energy scores for each position. The CBO model further calculates the logarithm of each calculated energy score. The CBO model further generates a score for the polypeptide by summing the logarithms of each calculated energy score, where the score represents a functional property of the polypeptide.

[0045] The present disclosure also provides a system including, among other things, a processor and a memory having computer code instructions stored thereon. The processor and memory, together with the computer code instructions, are configured to cause the system to score a polypeptide sequence using a CBO model. The CBO model calculates a plurality of energy scores for each amino acid position in the amino acid sequence of the polypeptide, where the plurality of energy scores is based on substituting an amino acid at a given position in the amino acid sequence with each of a plurality of different amino acids. The CBO model then adds each calculated energy score to an array. The CBO model then generates a normalized sum of the energy scores for each position. The CBO model then calculates the logarithm of each calculated energy score. The CBO model then generates a score for the polypeptide by summing the logarithms of each calculated energy score, where the score represents a functional property of the polypeptide.

[0046] Also provided herein are, inter alia, polypeptides that specifically bind to thymic stromal lymphopoietin (TSLP), the polypeptides comprising an amino acid sequence that is assigned a score above a predetermined threshold by a computational binding optimization (CBO) model when the amino acid sequence of the polypeptide is scored.

[0047] The present disclosure also provides, inter alia, polypeptides that bind to human thymic stromal lymphopoietin (TSLP), the polypeptides being designed by a method comprising generating a polypeptide sequence with a CBO model. In some embodiments, the polypeptides are further designed by validating the generated polypeptide sequence by calculating multiple energy scores for each amino acid position in the polypeptide sequence using the CBO model. The energy scores can be based on substituting each of multiple different amino acids for an amino acid at a given position in the polypeptide sequence. In some embodiments, the polypeptides are further validated by adding each calculated energy score to an array. In some embodiments, the polypeptides are further validated by generating a normalized sum of the energy scores for each position. In some embodiments, the polypeptides are further validated by calculating the logarithm of each calculated energy score. In some embodiments, the polypeptides are further validated by generating a score array for the polypeptide by summing the logarithms of each calculated energy score. This score represents a functional characteristic of the polypeptide's ability to bind to human TSLP. In some embodiments, a polypeptide sequence is validated if the polypeptide score is above a certain threshold.

[0048] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0049] The foregoing will become apparent from the following more detailed description of exemplary embodiments, as illustrated in the accompanying drawings, in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the embodiments.

[0050] In this figure, "Reference" refers to the reference antibody. [Brief explanation of the drawings]

[0051] [Figure 1] 1 depicts the amino acid sequence of human thymic stromal lymphopoietin (TSLP) (SEQ ID NO: 1). Epitope residues bound by the reference antibodies disclosed herein are indicated by asterisks. [Figure 2A] Figures 2A-2B illustrate the alignment of the heavy chain variable domain (VH) amino acid sequences of a reference antibody and exemplary antibodies AB-1 to AB-14. The heavy chain complementarity determining region (HCDR) amino acid sequence, as determined by ImMunoGeneTics (IMGT) numbering, is indicated using underlining. Bold indicates variable residues in the reference antibody and AB-1 to AB-14 (designated "Xn" throughout this disclosure). "*" indicates paratope residues. See also the VH consensus sequence (SEQ ID NO: 2) in Table 1. Paratope residues were defined as antibody residues in the reference antibody that are within 5 angstroms of the antigen when bound to human TSLP. [Figure 2B]Figures 2A-2B illustrate the alignment of the heavy chain variable domain (VH) amino acid sequences of a reference antibody and exemplary antibodies AB-1 to AB-14. The heavy chain complementarity determining region (HCDR) amino acid sequence, as determined by ImMunoGeneTics (IMGT) numbering, is indicated using underlining. Bold indicates variable residues in the reference antibody and AB-1 to AB-14 (designated "Xn" throughout this disclosure). "*" indicates paratope residues. See also the VH consensus sequence (SEQ ID NO: 2) in Table 1. Paratope residues were defined as antibody residues in the reference antibody that are within 5 angstroms of the antigen when bound to human TSLP. [Figure 3A] 3A-3B illustrate the alignment of the light chain variable domain (VL) amino acid sequences of the reference antibody and exemplary antibodies AB-1 to AB-14. The heavy chain complementarity determining region (LCDR) amino acid sequences, as determined by ImMunoGeneTics (IMGT) numbering, are indicated using underlining. Bold indicates variable residues in the reference antibody and AB-1 to AB-14 (designated "Xn" throughout this disclosure). See also the VL consensus sequence in Table 2 (SEQ ID NO: 18). [Figure 3B] 3A-3B illustrate the alignment of the light chain variable domain (VL) amino acid sequences of the reference antibody and exemplary antibodies AB-1 to AB-14. The heavy chain complementarity determining region (LCDR) amino acid sequences, as determined by ImMunoGeneTics (IMGT) numbering, are indicated using underlining. Bold indicates variable residues in the reference antibody and AB-1 to AB-14 (designated "Xn" throughout this disclosure). See also the VL consensus sequence in Table 2 (SEQ ID NO: 18). [Figure 4A] 4A-4B show binding profiles of non-limiting examples of anti-TSLP antibodies to human TSLP (hTSLP) generated using a direct enzyme-linked immunosorbent assay (ELISA). [Figure 4B]4A-4B show binding profiles of non-limiting examples of anti-TSLP antibodies to human TSLP (hTSLP) generated using a direct enzyme-linked immunosorbent assay (ELISA). [Figure 5A] 5A-5B show binding profiles of non-limiting examples of anti-TSLP antibodies to cynomolgus monkey TSLP (cynoTSLP) generated using direct ELISA. [Figure 5B] 5A-5B show binding profiles of non-limiting examples of anti-TSLP antibodies to cynomolgus monkey TSLP (cynoTSLP) generated using direct ELISA. [Figure 6A] 6A-6B show binding profiles of non-limiting examples of anti-TSLP antibodies to hTSLP generated using blocking ELISA. [Figure 6B] 6A-6B show binding profiles of non-limiting examples of anti-TSLP antibodies to hTSLP generated using blocking ELISA. [Figure 7A] Figures 7A-7B show binding profiles of non-limiting examples of anti-TSLP antibodies to cynoTSLP generated using blocking ELISA. [Figure 7B] Figures 7A-7B show binding profiles of non-limiting examples of anti-TSLP antibodies to cynoTSLP generated using blocking ELISA. [Figure 8A] Figures 8A-8C show non-limiting examples of anti-TSLP antibodies that neutralize the binding of hTSLP to the TSLP receptor (TSLPR). Ba / F3-STAT5-Luc cells expressing hTSLPR and hIL-7Rα, STAT5 reporter cells, were used to assess the binding between hTSLP and TSLPR based on hTSLP-induced STAT5 activity. Figure 8A shows TSLP-mediated STAT5 signaling as a function of hTSLP concentration. Figures 8B-8C show the effect of anti-TSLP antibodies on TSLP-mediated STAT5 signaling. [Figure 8B]Figures 8A-8C show non-limiting examples of anti-TSLP antibodies that neutralize the binding of hTSLP to the TSLP receptor (TSLPR). Ba / F3-STAT5-Luc cells expressing hTSLPR and hIL-7Rα, STAT5 reporter cells, were used to assess the binding between hTSLP and TSLPR based on hTSLP-induced STAT5 activity. Figure 8A shows TSLP-mediated STAT5 signaling as a function of hTSLP concentration. Figures 8B-8C show the effect of anti-TSLP antibodies on TSLP-mediated STAT5 signaling. [Figure 8C] Figures 8A-8C show non-limiting examples of anti-TSLP antibodies that neutralize the binding of hTSLP to the TSLP receptor (TSLPR). Ba / F3-STAT5-Luc cells expressing hTSLPR and hIL-7Rα, STAT5 reporter cells, were used to assess the binding between hTSLP and TSLPR based on hTSLP-induced STAT5 activity. Figure 8A shows TSLP-mediated STAT5 signaling as a function of hTSLP concentration. Figures 8B-8C show the effect of anti-TSLP antibodies on TSLP-mediated STAT5 signaling. [Figure 9A] 9A-9B are graphs showing that exemplary anti-TSLP antibodies block TSLP binding to peripheral blood-derived human myeloid dendritic cells (mDCs). The curves show the effect of anti-TSLP antibodies on TSLP-induced CCL17 expression by mDCs from one representative donor. [Figure 9B] 9A-9B are graphs showing that exemplary anti-TSLP antibodies block TSLP binding to peripheral blood-derived human myeloid dendritic cells (mDCs). The curves show the effect of anti-TSLP antibodies on TSLP-induced CCL17 expression by mDCs from one representative donor. [Figure 10A]Figures 10A-10D are graphs showing binding profiles of AB-2a, AB-2b, a reference antibody, an IgG1 isotype control, and an IgG2 isotype control generated using direct ELISA. Figure 10A shows the binding profile for hTSLP, Figure 10B shows the binding profile for cynoTSLP, Figure 10C shows the binding profile for mouse TSLP, and Figure 10D shows the binding profile for human IL-7. [Figure 10B] Figures 10A-10D are graphs showing binding profiles of AB-2a, AB-2b, a reference antibody, an IgG1 isotype control, and an IgG2 isotype control generated using direct ELISA. Figure 10A shows the binding profile for hTSLP, Figure 10B shows the binding profile for cynoTSLP, Figure 10C shows the binding profile for mouse TSLP, and Figure 10D shows the binding profile for human IL-7. [Figures 10C-10D] Figures 10A-10D are graphs showing binding profiles of AB-2a, AB-2b, a reference antibody, an IgG1 isotype control, and an IgG2 isotype control generated using direct ELISA. Figure 10A shows the binding profile for hTSLP, Figure 10B shows the binding profile for cynoTSLP, Figure 10C shows the binding profile for mouse TSLP, and Figure 10D shows the binding profile for human IL-7. [Figure 11A] Figures 11A-11B are graphs showing Ba / F3 assay results for AB-2a, AB-2b, a reference antibody, an IgG1 isotype control, and an IgG2 isotype control. Figure 11A shows human TSLPR / IL-7Rα Ba / F3 assay results. Figure 11B shows cynoTSLPR / IL-7Rα Ba / F3 assay results. [Figure 11B]Figures 11A-11B are graphs showing Ba / F3 assay results for AB-2a, AB-2b, a reference antibody, an IgG1 isotype control, and an IgG2 isotype control. Figure 11A shows human TSLPR / IL-7Rα Ba / F3 assay results. Figure 11B shows cynoTSLPR / IL-7Rα Ba / F3 assay results. [Figure 12A] 10 illustrates zero-order (1002), first-order (1004), and second-order (1006) models. [Figure 12B] FIG. 1 is a flow diagram illustrating an example of a process used by embodiments of the present disclosure to determine a claimed sequence. [Figure 13] 1 illustrates an exemplary computer network or similar digital processing environment in which embodiments of the present invention may be implemented. [Figure 14] 14 is a diagram of an exemplary internal structure of a computer (e.g., a client processor / device or a server computer) in the computer system of FIG. 13. DETAILED DESCRIPTION OF THE INVENTION

[0052] A description of exemplary embodiments follows.

[0053] Some aspects of the present disclosure are described below with reference to examples that are for illustrative purposes only. It should be understood that numerous specific details, relationships, and methods are set forth to provide a thorough understanding of the present disclosure. However, one of ordinary skill in the art will readily recognize that the present disclosure can be practiced without one or more of the specific details, or with other methods, protocols, reagents, cell lines, and animals. The present disclosure is not limited to the illustrated order of acts or events, as some acts may occur in a different order and / or simultaneously with other acts or events. Furthermore, not all illustrated acts, steps, or events are required to achieve the events of the present disclosure. Many of the techniques and procedures described or referenced herein are well understood and commonly employed by those of ordinary skill in the art using conventional methodology.

[0054] definition Unless otherwise defined, all technical terms, notation, and other scientific terms or terminology used herein are intended to have the meaning commonly understood by one of ordinary skill in the art to which this disclosure pertains. In some cases, terms having a commonly understood meaning are defined herein for clarity and / or ease of reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial difference from that generally understood in the art. Furthermore, it will be understood that terms, such as those defined in commonly used dictionaries, should be construed to have a meaning consistent with their meaning in the context of the relevant art and / or as otherwise defined herein.

[0055] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0056] When referring to elements disclosed herein, the articles "a," "an," "the," and "said" are intended to mean that there are one or more of the elements. Moreover, the one or more elements may be the same or different. For example, unless the context clearly dictates otherwise, reference to "a polypeptide" includes a single polypeptide as well as two or more polypeptides.

[0057] Throughout this specification and the claims that follow, unless the context otherwise requires, the term "comprise," and variations such as "comprises" and "comprising," will be understood to imply, for example, the inclusion of a stated integer or step or group of integers or steps, but not the exclusion of any other integer or step or group of integers or steps. As used herein, the term "comprising" can be replaced by the terms "containing" or "including."

[0058] As used herein, the term "consisting of" excludes any element, step, or ingredient not specified in the claim element. As used herein, the term "consisting essentially of" does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claim.

[0059] Also provided herein are corresponding embodiments for any and all embodiments characterized by the terms "comprising," "containing," "including," or "having," where these terms are replaced with the terms "consisting of" and / or "consisting essentially of."

[0060] As used herein, the coordinating term "and / or" between multiple listed elements is understood to encompass both individual and combined options. For example, when two elements are coordinating "and / or," the first option indicates that the first element is applicable without the second element. The second option indicates that the second element is applicable without the first element. The third option indicates that the first and second elements are applicable together. Any one of these options is understood to fall within the meaning and thus meet the requirements of the term "and / or" as used herein. The simultaneous applicability of two or more options also falls within the meaning and thus meets the requirements of the term "and / or."

[0061] It should be understood that for all numerical boundaries describing certain parameters in this application, such as "about," "at least," "less than," "less than," and "greater than," the description necessarily also encompasses any range bounded by the stated values. Thus, for example, the description "at least 1, 2, 3, 4, or 5" also describes the ranges 1-2, 1-3, 1-4, 1-5, 2-3, 2-4, 2-5, 3-4, 3-5, and 4-5, among others.

[0062] When lists are presented, unless otherwise stated, it is to be understood that each individual element of that list, and any combination of that list, is a separate embodiment. For example, a list of embodiments presented as "A, B, or C" should be interpreted to include the embodiments "A," "B," "C," "A or B," "A or C," "B or C," or "A, B, or C."

[0063] As used herein, the term "about" means that a particular value falls within an acceptable error range as determined by one of ordinary skill in the art. Typically, the acceptable error range for a particular value depends, at least in part, on how the value is measured or determined, e.g., the limitations of the measurement system. For example, "about" can mean within an acceptable standard deviation, as practiced in the art. Alternatively, "about" can mean within a range of ±20%, e.g., ±10%, ±5%, or ±1% from the given value. It should be understood that the term "about" can precede any specific value specified herein, except for the specific values used in the examples. When "about" precedes a range, such as "90-99.9%," the term "about" should be read to apply to both of the given values in the range; thus, "about 90-99.9%" means approximately 90% to approximately 99.9%.

[0064] As used herein, the term "polypeptide" refers to a polymer of at least two amino acids covalently linked by amide bonds, regardless of length or post-translational modification (e.g., glycosylation or phosphorylation). Polypeptides can contain any suitable L- and / or D-amino acids, including conventional α-amino acids (e.g., alanine, glycine, valine), non-α-amino acids (e.g., β-alanine, 4-aminobutyric acid, 6-aminocaproic acid, sarcosine, statins), and unusual amino acids (e.g., citrulline, homocitrulline, homoserine, norleucine, norvaline, ornithine). Amino, carboxyl, and / or other functional groups on a polypeptide can be free (e.g., unmodified) or protected with suitable protecting groups. Suitable protecting groups for amino and carboxyl groups, and methods for adding or removing protecting groups, are known in the art and are disclosed, for example, in Green and Wuts, "Protecting Groups in Organic Synthesis," John Wiley and Sons, 1991. Functional groups of polypeptides may also be derivatized (e.g., alkylated) or labeled (e.g., with a detectable label, such as a fluorogenic agent or hapten) using methods known in the art. Polypeptides may optionally contain one or more modifications (e.g., amino acid linkers, acylation, acetylation, amidation, methylation, terminal modifiers (e.g., cyclization modifications), N-methyl-α-amino group substitutions). In addition, polypeptides may be analogs of known and / or naturally occurring peptides, for example, peptide analogs having one or more conservative amino acid residue substitutions.

[0065] As used herein, a "polynucleotide" is defined as a plurality of nucleotides and / or nucleotide analogs linked together into a single molecule. In some embodiments, polynucleotides disclosed herein comprise deoxyribonucleotides. In some embodiments, polynucleotides comprise ribonucleotides. Non-limiting examples of polynucleotides include single-stranded, double-stranded, or multi-stranded DNA or RNA, DNA-RNA hybrids (e.g., each "T" position may be independently substituted with "U" or vice versa), or polymers containing purine and pyrimidine bases, or other natural, chemically or biochemically modified, non-natural, or derivatized nucleotide bases. The backbone of a polynucleotide can comprise sugar and phosphate groups, modified or substituted sugar or phosphate groups, polymers of synthetic subunits such as phosphoramidates, or combinations thereof.

[0066] As used herein, the term "sequence identity" refers to the percentage of identical residues in two nucleotide sequences when the sequences are aligned to maximize identity. For sequence alignment and comparison, typically one sequence is designated as a reference sequence to which a test sequence is compared. The sequence identity between a reference sequence and a test sequence is expressed as the percentage of positions across the entire length of the reference sequence that the reference sequence and the test sequence share the same nucleotide or amino acid when aligned to maximize identity. For example, two sequences are considered to have 70% sequence identity when the test sequence has the same nucleotide residues at 70% of the same positions across the entire length of the reference sequence when aligned to maximize identity.

[0067] Alignment of sequences for comparison to achieve the highest level of identity can be readily performed by one of skill in the art using an appropriate alignment method or algorithm, and in some cases, the alignment may include gaps introduced to provide the highest level of identity. Examples include the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), the homology algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), the similarity search method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), and visual inspection (see generally Ausubel et al., Current Protocols in Molecular Biology).

[0068] When using a sequence comparison algorithm, test and reference sequences are input into a computer, followed by designating coordinates as necessary and designating sequence algorithm program parameters. The sequence comparison algorithm then calculates the percent sequence identity for one or more test sequences relative to the reference sequence based on the designated program parameters. A commonly used tool for determining percent sequence identity is the Protein Basic Local Alignment Search Tool (BLASTP) (Altschul et al., 1990), available from the National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health.

[0069] As used herein, the term "substantially similar" refers to a polypeptide disclosed herein that has substantially similar amino acid sequence (e.g., at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% amino acid residues share amino acid sequence identity) and that substantially retains one or more functional properties of the specified polypeptide disclosed herein (e.g., AB-1). In some embodiments, the one or more functional properties are selected from, without limitation, substantially similar binding affinity, substantially similar binding specificity, substantially similar inhibitory activity, substantially similar neutralizing activity, and substantially similar self-association properties.

[0070] As used herein, "complementarity-determining region (CDR)" encompasses any CDR defined by art-recognized methods for identifying CDR residues on an antibody. See, e.g., Kabat, E.A., et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242; Chothia et al., (1989) Nature 342:877; Chothia, C. et al., (1987) J. Mol. Biol. 196:901-917; Al-lazikani et al., (1997) J. Mol. Biol. 273:927-948; and Almagro, J. Mol. Recognit. 17:132-143 (2004). See also hgmp.mrc.ac.uk and bioinf.org.uk / abs. Two antibodies are determined to have the same CDRs as each other when the identities of the CDRs for both antibodies are determined using the same method for CDRs based on HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and / or LCDR3.

[0071] The framework regions and CDR extents of an antibody can be identified using any one of several suitable methodologies known in the art, for example, the Kabat definition, the Chothia definition, the AbM definition, and / or the contact definition. Publicly available and / or commercially available tools for identifying framework and / or CDR regions include IgBlast (accessible at www.ncbi.nlm.nih.gov / igblast / ), Scaligner (available from drugdesigntech at www.scaligner.com / ), the IMGT rules and / or tools (see, e.g., www.imgt.org / IMGTScientificChart / Nomenclature / IMGT-FRCDRdefinition.html, also accessible at www.imgt.org / ), the Chothia Canonical Examples of such information include the Antigen Receptor Numbering and Receptor Classification (ANARCI, available at opig.stats.ox.ac.uk / webapps / newsabdab / sabpred / anarci / ), the Antigen Receptor Assignment (available at www.bioinf.org.uk / abs / chothia.html), the Antigen Receptor Numbering and Receptor Classification (ANARCI, available at opig.stats.ox.ac.uk / webapps / newsabdab / sabpred / anarci / ), or the Paratome web server (available at www.ofranlab.org / paratome / ; see Vered Kunik, et al, Nucleic Acids Research, Volume 40, Issue W1, 1 July 2012, Pages W521-W524).

[0072] For example, for AB-2, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 amino acid sequences include (1) SEQ ID NO:31, SEQ ID NO:35, SEQ ID NO:50, SEQ ID NO:60, SEQ ID NO:67, and SEQ ID NO:71, respectively, when determined by IMGT numbering; (2) SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, and SEQ ID NO:71, respectively, when determined by Kabat numbering; and (3) SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, and SEQ ID NO:71, respectively, when determined by Chothia numbering.

[0073] As used herein, the term "paratope" refers to a set of amino acid residues in an antibody or antigen-binding fragment thereof that contribute to binding interactions with an epitope of a target protein. The binding interactions can be hydrogen bonds, salt bridges, van der Waals interactions, ionic bonds, or a combination thereof. The binding interactions can be direct or indirect, e.g., via a coordinating intermediate molecule such as an ion or water. The residues of the paratope, in some embodiments, include only residues that are part of a defined CDR. In other embodiments, the residues of the paratope further include one or more residues that are not part of a defined CDR.

[0074] As used herein, the term "antibody mimetics" refers to polypeptides that have the ability to mimic the antigen-binding ability of antibodies but are structurally distinct from natural antibody structures. Examples of antibody mimetics include, but are not limited to, adnectins, affibodies, affilins, affimers, affitins, alphabodies, anticalins, avimers, DARPins, fynomers, Kunitz domain peptides, monobodies, nanobodies, nanoCLAMPs, and versabodies.

[0075] As used herein, the term "K D The "binding constant," also known as the "equilibrium dissociation constant," or "affinity constant," is a measure of the reversible association between two molecular species (e.g., an antibody and a target protein) and includes both actual and apparent binding affinity. Binding affinity can be determined using methods known in the art, including, for example, surface plasmon resonance measurements using, for example, biolayer interferometry (Octet, ForteBio) or surface plasmon resonance (Biacore) systems and assays. A reference comparing various surface technologies for measuring binding affinity and kinetics is Yang, D., Singh, A., Wu, H., & Kroe-Barrett, R., Comparison of biosensor platforms in the evaluation of high affinity antibody-antigen binding kinetics, Analytical Biochemistry 508:78-96 (2016), the contents of which are incorporated herein by reference in their entirety.

[0076] The term "subject" or "patient" refers to an animal (e.g., a mammal, such as a human) diagnosed with or suspected of having a TSLP-associated disease or condition (e.g., a disease or condition associated with dysregulated TSLP expression, such as asthma or atopic dermatitis (AD)), or an animal at risk for developing such a condition. Diagnosis may be performed by any method or technique known in the art. One of skill in the art will understand that subjects for treatment according to the present disclosure may have undergone standard testing, or may be identified as at risk without testing because of the presence of one or more risk factors associated with the disease or condition.

[0077] The term "pharmaceutically acceptable" means that the substance or composition it modifies is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and is commensurate with a reasonable benefit-risk ratio.

[0078] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that, within the scope of sound medical judgment, is suitable for use in contact with mammalian tissues without undue toxicity, irritation, allergic reaction, etc., and is commensurate with a reasonable benefit-risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19 (the relevant teachings of which are incorporated herein by reference in their entirety). Pharmaceutically acceptable salts of the drugs / compounds described herein include salts derived from suitable inorganic and organic acids and suitable inorganic and organic bases.

[0079] Examples of salts derived from suitable acids include salts of amino groups formed with inorganic acids such as hydrochloric, hydrobromic, phosphoric, sulfuric and perchloric acids, or with organic acids such as acetic, oxalic, maleic, tartaric, citric, succinic or malonic acid, or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts derived from suitable acids include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cinnamate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, glutarate, glycolate, hemisulfate, heptanoate, hexanoate, hydroiodide, hydroxybenzoate, 2-hydroxy-ethanesulfonate. , hydroxymaleate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 2-phenoxybenzoate, phenylacetate, 3-phenylpropionate, phosphate, pivalate, propionate, pyruvate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.

[0080] Either the mono-, di-, or tri-acid salts can be formed, and such salts can exist in either a hydrated, solvated or substantially anhydrous form.

[0081] Salts derived from appropriate bases include those derived from inorganic bases such as alkali metal, alkaline earth metal, and ammonium bases, as well as those derived from aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine, and picoline, or N +Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, barium, and the like. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed, where appropriate, using counterions such as halides, hydroxides, carboxyls, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0082] "Pharmaceutically acceptable carrier" refers to a non-toxic carrier or excipient that does not destroy the pharmacological activity of the drug with which it is formulated and is non-toxic when administered in a dose sufficient to deliver a therapeutic amount of the drug. Pharmaceutically acceptable carriers that can be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol, and wool fat.

[0083] "Treating" or "treatment," as used herein, refers to delivering a treatment to a subject, such as a mammal, in need thereof (e.g., by administering one or more therapeutic agents to the mammal). "Treating" or "treatment" includes inhibiting a disease or condition (e.g., by slowing or stopping its progression or by causing regression of the disease or condition), as well as alleviating the symptoms caused by the disease or condition.

[0084] The term "treating" or "treatment" refers to the medical management of a subject with the intent to reverse, improve, stabilize (i.e., not worsen), prevent, or cure a disease, pathological condition, or disorder, such as the specific indications exemplified herein. This term includes active treatment (treatment directed at reversing the disease, pathological condition, or disorder), causal treatment (treatment directed at the cause of the associated disease, pathological condition, or disorder), palliative treatment (treatment designed to relieve symptoms), preventive treatment (treatment directed at minimizing or partially or completely preventing the occurrence of the associated disease, pathological condition, or disorder), and supportive treatment (treatment used to complement another treatment). Treatment also includes the reduction of the extent of the disease or condition, whether detectable or undetectable; preventing the spread of the disease or condition; slowing or slowing the progression of the disease or condition; improvement or palliation of the disease or condition; and remission (whether partial or total). "Ameliorating" or "alleviating" a disease or condition means lessening the severity and / or undesirable clinical symptoms of the disease, disorder, or condition and / or slowing or prolonging its progression over time compared to the severity or course of time that would occur in the absence of treatment. "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the condition or disorder, as well as those prone to the condition or disorder or those in whom the condition or disorder is to be prevented.

[0085] A "pharmaceutical composition" refers to a formulation of one or more therapeutic agents and a vehicle accepted in the art for delivery of biologically active agents to a subject, e.g., a human. In some embodiments, a pharmaceutical composition may include one or more pharmaceutically acceptable excipients, diluents, or carriers. In some embodiments, a pharmaceutical composition suitable for use in the methods disclosed herein further includes one or more pharmaceutically acceptable carriers.

[0086] A "pharmaceutically acceptable carrier, diluent, or excipient" includes any adjuvant, carrier, excipient, flow agent, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier approved by the U.S. Food and Drug Administration as acceptable for human or veterinary use.

[0087] A "pharmaceutically acceptable carrier" refers to a component of a pharmaceutical composition, other than the active ingredient, that is nontoxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives. In some embodiments, a carrier can be a diluent, adjuvant, excipient, or vehicle with which a drug (e.g., a polynucleotide) is administered. Such vehicles can be liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. For example, 0.4% saline and 0.3% glycine can be used. Such solutions are sterile and generally free of particulate matter. They can be sterilized by conventional, well-known sterilization techniques (e.g., filtration). The compositions can contain pharmaceutically acceptable auxiliary substances as needed to approximate physiological conditions, such as pH adjusting and buffering agents, stabilizers, thickening agents, lubricating agents, and coloring agents. The concentration of drug in such pharmaceutical formulations can vary widely, i.e., from less than about 0.5% to at least about 1%, or as high as 15% or 20%, 25%, 30%, 35%, 40%, 45%, or 50% by weight. The concentration will be selected according to the mode of administration, based primarily on the required dose, fluid volume, viscosity, etc. Suitable vehicles and formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 21, pp. 111-115, 1997, including other human proteins, e.g., human serum albumin. st Edition, Troy, DB ed., Lipincott Williams and Wilkins, Philadelphia, PA 2006, Part 5, Pharmaceutical Manufacturing: 691-1092 (e.g., pages 958-89).

[0088] Non-limiting examples of pharmaceutically acceptable carriers include solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible, such as salts, buffers, antioxidants, saccharides, aqueous or non-aqueous carriers, preservatives, wetting agents, surfactants or emulsifying agents, or combinations thereof.

[0089] Non-limiting examples of buffers are acetate, citrate, formate, succinate, phosphate, carbonate, malate, aspartate, histidine, borate, Tris buffer, HEPPSO, and HEPES.

[0090] Non-limiting examples of antioxidants are ascorbic acid, methionine, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, lecithin, citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, and tartaric acid.

[0091] Non-limiting examples of amino acids are histidine, isoleucine, methionine, glycine, arginine, lysine, L-leucine, trileucine, alanine, glutamic acid, L-threonine, and 2-phenylamine.

[0092] Non-limiting examples of surfactants include polysorbates (e.g., polysorbate-20 or polysorbate 80); poloxamers (e.g., poloxamer 188); Triton; sodium octyl glycoside; lauryl sulfobetaine, myristyl sulfobetaine, linoleyl sulfobetaine, or stearyl sulfobetaine; lauryl sarcosine, myristyl sarcosine, linoleyl sarcosine, or stearyl sarcosine; linoleyl betaine, myristyl betaine, or cetyl betaine; lauroamidopropyl betaine, cocamidopropyl betaine, linoleamidopropyl betaine, myristamidopropyl betaine, palmidopropyl betaine, or isostearamidopropyl betaine (e.g., lauroamidopropyl); myristamidopropyl dimethylamine, palmidopropyl dimethylamine, or isostearamidopropyl dimethylamine; sodium methyl cocoyl taurate, or disodium methyl oleyl taurate; and the MONAQUA™ series (Mona Industries, Inc., Paterson, NJ), copolymers of polyethyl glycol, polypropyl glycol, and ethylene and propylene glycol (e.g., PLURONICS™, PF68, etc.).

[0093] Non-limiting examples of preservatives are phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrite, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride, alkyl parabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof.

[0094] Non-limiting examples of saccharides are monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, such as glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, dextran, erythritol, glycerol, arabitol, sylitol, sorbitol, mannitol, melibiose, melezitose, raffinose, mannotriose, stachyose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, or iso-maltulose.

[0095] Non-limiting examples of salts include acid addition salts and base addition salts. Acid addition salts include those derived from non-toxic inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphorous acid, as well as those derived from non-toxic organic acids such as aliphatic monocarboxylic and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, and aliphatic and aromatic sulfonic acids. Base addition salts include those derived from alkaline earth metals such as sodium, potassium, magnesium, and calcium, as well as those derived from non-toxic organic amines such as N,N'-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, and procaine. In some embodiments, the salt is sodium chloride (NaCl).

[0096] The agents (e.g., polynucleotides) described herein may be prepared in accordance with standard procedures and administered in dosages selected to reduce, prevent, or eliminate the condition under treatment, or to slow or stop its progression (for a review of methods for administering various agents for human therapy, see, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, and Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, McGraw-Hill, New York, NY, the contents of which are incorporated herein by reference).

[0097] "Administering" or "administration," as used herein, refers to providing a compound, composition, or pharmaceutically acceptable salt thereof described herein to a subject in need of treatment or prevention. Administering can be performed, for example, once, multiple times, and / or for one or more durations. Administering includes direct administration (including self-administration) and indirect administration (including the act of prescribing a drug or instructing a subject to take a drug). For example, as used herein, a person (e.g., a physician) who instructs a subject (e.g., a human patient) to self-administer a drug (e.g., a medication) or to have another person administer a medication and / or who prescribes a drug to a patient is administering a medication to the subject.

[0098] A "therapeutically effective amount," "effective amount," or "effective dosage" refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result (e.g., treating, curing, inhibiting, or ameliorating a physiological response or condition, etc.). The full therapeutic effect may not occur by administration of one dose, but may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. A therapeutically effective amount may vary depending on factors such as the disease state, age, sex, and weight of the mammal (e.g., a human patient), the mode of administration, and the ability of the therapeutic agent or combination of therapeutic agents to elicit the desired response.

[0099] The effective amount of a drug to be administered can be determined by a skilled clinician using the guidance provided herein and other methods known in the art. Relevant factors include the drug being administered, the pharmaceutical formulation, the route of administration, the type of disease or disorder, and the identity (e.g., age, sex, weight) or host of the subject being treated. For example, a suitable dosage may be about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 10 mg / kg, or about 0.01 mg / kg to about 1 mg / kg of body weight per treatment. Determining the dosage for a particular drug, subject, and disease is well within the capabilities of one of ordinary skill in the art. Preferably, the dosage does not cause adverse side effects or causes minimal adverse side effects.

[0100] The desired response or desired outcome can include an effect at the cellular level, a tissue level, or a clinical result. Thus, the term "therapeutically effective amount" or its equivalents depends on the context in which it is applied. For example, in some embodiments, it is an amount of a composition sufficient to achieve a therapeutic response compared to the response obtained in the absence of administration of the composition. In other embodiments, it is an amount that produces a beneficial or desired outcome in a subject compared to a control. As defined herein, a therapeutically effective amount of a composition (e.g., a pharmaceutical composition) disclosed herein can be readily determined by one of ordinary skill in the art using routine methods known in the art. The dosage regimen and route of administration can be adjusted to provide an optimal therapeutic response.

[0101] Thymic stromal lymphopoietin (TSLP) As used herein, TSLP includes wild-type TSLP protein (e.g., wild-type human TSLP protein or a homolog thereof) and truncated forms, mutants, and engineered versions thereof, of full-length and truncated TSLP protein, and modified forms (e.g., post-translationally modified forms) of full-length and truncated TSLP protein.

[0102] A non-limiting example of a human TSLP sequence is GenBank: AAK67490.1 (SEQ ID NO: 1). [ka]

[0103] A non-limiting example of a cynomolgus monkey TSLP sequence is GenBank: EHH54440.1 (SEQ ID NO: 83). [ka]

[0104] In some embodiments, the polypeptide binds to a wild-type TSLP protein (e.g., a TSLP protein having the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 83, or both). In some embodiments, the polypeptide binds to a TSLP protein comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the polypeptide binds to a TSLP protein comprising the amino acid sequence of SEQ ID NO: 1 and a TSLP protein comprising the amino acid sequence of SEQ ID NO: 83.

[0105] In some embodiments, the polypeptide binds to a mutant or engineered TSLP protein. In some embodiments, the mutant or engineered TSLP protein comprises an amino acid sequence having at least about 90% sequence identity to a wild-type TSLP protein, e.g., at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% sequence identity to a wild-type TSLP protein (e.g., SEQ ID NO: 1 or SEQ ID NO: 83). In some embodiments, the mutant or engineered TSLP protein comprises an amino acid sequence having about 90-99.9%, 90-99.8%, 92-99.8%, 92-99.6%, 94-99.6%, 94-99.5%, 95-99.5%, 95-99.4%, 96-99.4%, 96-99.2%, 97-99.2%, or 97-99% sequence identity to a wild-type TSLP protein.

[0106] In some embodiments, the polypeptide binds to a modified TSLP protein.

[0107] In some embodiments, the polypeptide is capable of binding to one or more epitope residues in a TSLP protein (e.g., full-length human TSLP), e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 epitope residues of a TSLP protein. In some embodiments, the polypeptide is capable of binding to one or more epitope residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all 15 residues) selected from N64, N65, T66, V67, S68, C69, S70, N71, R72, H74, C75, E78, R150, F151, and R153 of SEQ ID NO:1.

[0108] In some embodiments, the polypeptide binds to one or more epitope residues of a TSLP protein (e.g., one or more epitope residues of SEQ ID NO: 1). In some embodiments, the polypeptide binds to one or more epitope residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all 15 residues) selected from N64, N65, T66, V67, S68, C69, S70, N71, R72, H74, C75, E78, R150, F151, and R153 of SEQ ID NO: 1.

[0109] Control polypeptide As used herein, the term "control agent" or "control agent polypeptide" refers to a polypeptide (e.g., an immunoglobulin molecule) that specifically binds to a TSLP protein and is not a polypeptide disclosed herein. Comparison of the sequences of a control agent polypeptide and a polypeptide disclosed herein can reveal structural differences therebetween (e.g., differences at one or more amino acid positions, such as amino acid substitutions). Because the polypeptides disclosed herein have more than insubstantial differences (e.g., one or more substantial differences) compared to the control agent polypeptide, the polypeptides disclosed herein will, under controlled conditions, exhibit one or more (i.e., one, two, or all three) of a different function, a different appearance, or a different effect achieved compared to the control agent polypeptide. A reference polypeptide can differ from a polypeptide disclosed herein by one or more amino acids, for example, in some embodiments, by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acids. In some embodiments, a reference polypeptide deviates from a polypeptide disclosed herein by at least about 0.4%, 0.8%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% or more amino acid percent identity.

[0110] In some embodiments, the control agent polypeptide is: a) the heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2) and heavy chain complementarity determining region 3 (HCDR3) sequences of SEQ ID NO: 31, SEQ ID NO: 33 and SEQ ID NO: 48, respectively; b) the light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2) and light chain complementarity determining region 3 (LCDR3) sequences of SEQ ID NO: 59, SEQ ID NO: 67 and SEQ ID NO: 69, respectively; or Both a) and b) The antibody comprises:

[0111] In some embodiments, the control agent polypeptide is: a) the HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO: 31, SEQ ID NO: 33 and SEQ ID NO: 48, respectively; and b) LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO: 59, SEQ ID NO: 67 and SEQ ID NO: 69, respectively The antibody comprises:

[0112] For SEQ ID NOs: 31, 33 and 48, see Table 3 and Figure 2. For SEQ ID NOs: 59, 67 and 69, see Table 3 and Figure 3.

[0113] In some embodiments, the control agent polypeptide is: a) an immunoglobulin heavy chain variable region (V) comprising the amino acid sequence of SEQ ID NO: 3; H )domain; b) an immunoglobulin light chain variable region (V) comprising the amino acid sequence of SEQ ID NO: 19; L ) domain, or Both a) and b) The antibody comprises:

[0114] In some embodiments, the control agent polypeptide is: a) an immunoglobulin heavy chain variable region (V) comprising the amino acid sequence of SEQ ID NO: 3; H ) domain; and b) an immunoglobulin light chain variable region (V) comprising the amino acid sequence of SEQ ID NO: 19;L )domain The antibody comprises:

[0115] For SEQ ID NO: 3, see Table 1 and Figure 2. For SEQ ID NO: 19, see Table 2 and Figure 3.

[0116] In some embodiments, the control polypeptide is an antibody referred to herein as a "reference antibody." A reference antibody is an antibody that: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 81; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 82 Includes.

[0117] For SEQ ID NO: 81, see Table 4. For SEQ ID NO: 82, see Table 5.

[0118] The reference antibody binds to and neutralizes TSLP.

[0119] Polypeptides In some embodiments, the polypeptide specifically binds to TSLP. In some embodiments, the polypeptide is an antibody or an antigen-binding fragment thereof.

[0120] variable domain In some embodiments, the polypeptide (e.g., an antibody or antigen-binding fragment thereof) comprises an immunoglobulin heavy chain variable region (V H ), immunoglobulin light chain variable region (V L In some embodiments, the polypeptide comprises a V that has less than 100% sequence identity with the amino acid sequence of SEQ ID NO:3. H , V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 19 L , or both.

[0121] In some embodiments, the polypeptide is V H (e.g., rodent (e.g., mouse) V H , primate (e.g., human) V HMammal V H In some embodiments, the polypeptide comprises a V that is humanized (e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% humanized), includes human framework regions, or both. H Includes.

[0122] In some embodiments, the polypeptide has a V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO:3. H Includes.

[0123] In some embodiments, the polypeptide has a V having at least about 70% sequence identity to the amino acid sequence of SEQ ID NO:3. H In some embodiments, the polypeptide comprises a V having at least about 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO:3. H In some embodiments, the polypeptide comprises a V having at least about 85% sequence identity to the amino acid sequence of SEQ ID NO:3. H In some embodiments, the polypeptide comprises a V having at least about 90% sequence identity to the amino acid sequence of SEQ ID NO:3. H Includes.

[0124] In some embodiments, the polypeptide comprises a V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative substitution) compared to the amino acid sequence of SEQ ID NO:3. HFor example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a V that contains about 1-10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO:3. H In some embodiments, at least one amino acid substitution replaces only a heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2), and / or heavy chain complementarity determining region 3 (HCDR3) residue of SEQ ID NO: 3. In some embodiments, at least one amino acid substitution replaces only a non-CDR residue of SEQ ID NO: 3 (e.g., within a framework region).

[0125] In some embodiments, the amino acid substitution is a conservative substitution. The term "conservative amino acid substitution" or "conservative substitution" refers to an amino acid substitution having a BLOSUM62 value of 0 or greater.

[0126] In some embodiments, the amino acid substitution is a highly conservative substitution. The term "highly conservative amino acid substitution" or "highly conservative substitution" refers to an amino acid substitution having a value of at least 1 (e.g., at least 2) in BLOSUM62.

[0127] In some embodiments, the polypeptide has a V that has 100% sequence identity to the amino acid sequence of SEQ ID NO:3. H In some embodiments, the polypeptide comprises a V comprising the amino acid sequence of SEQ ID NO:3. H Includes.

[0128] In some embodiments, the polypeptide has a V having at least about 70% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-17. H The sequences identified as SEQ ID NOs: 4 to 17 are shown in Table 1 and are representative of human VH In some embodiments, the polypeptide corresponds to a V domain having at least about 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-17. H In some embodiments, the polypeptide comprises a V having at least about 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-17. H In some embodiments, the polypeptide comprises a V having at least about 90% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-17. H Includes.

[0129] In some embodiments, the polypeptide comprises a V that contains at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 4-17. H For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a V H Includes.

[0130] In some embodiments, the polypeptide has a V having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-17. H In some embodiments, the polypeptide comprises a V comprising the amino acid sequence of any one of SEQ ID NOs: 4-17. H Includes.

[0131] In some embodiments, the polypeptide has a V having at least about 70% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-7. H In some embodiments, the polypeptide comprises a V having at least about 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-7. H In some embodiments, the polypeptide comprises a V having at least about 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-7. H In some embodiments, the polypeptide comprises a V having at least about 90% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 4-7. H Includes.

[0132] In some embodiments, the polypeptide comprises a V that contains at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 4-7. H For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a V that contains about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 4-7. H Includes.

[0133] In some embodiments, the polypeptide has a V having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 4-7. H In some embodiments, the polypeptide comprises a V comprising the amino acid sequence of any one of SEQ ID NOs: 4-7. H Includes.

[0134] In some embodiments, the polypeptide is V L (e.g., rodent (e.g., mouse) V L , primate (e.g., human) V L Mammal V L In some embodiments, the polypeptide is humanized (e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% humanized). L or comprises human framework regions, or both.

[0135] In some embodiments, the polypeptide has a V that has less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 19. L Includes.

[0136] In some embodiments, the polypeptide has a V having at least about 70% sequence identity to the amino acid sequence of SEQ ID NO:19. L In some embodiments, the polypeptide comprises a V having at least about 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 19. L In some embodiments, the polypeptide comprises a V having at least about 85% sequence identity to the amino acid sequence of SEQ ID NO:19. L In some embodiments, the polypeptide comprises a V having at least about 90% sequence identity to the amino acid sequence of SEQ ID NO:19. L Includes.

[0137] In some embodiments, the polypeptide comprises a V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative substitution) compared to the amino acid sequence of SEQ ID NO: 19. LFor example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a V that contains about 1-10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 19. L In some embodiments, at least one amino acid substitution replaces only a light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2), and / or light chain complementarity determining region 3 (LCDR3) residue of SEQ ID NO: 19. In some embodiments, at least one amino acid substitution replaces only a non-CDR residue of SEQ ID NO: 19 (e.g., within a framework region).

[0138] In some embodiments, the amino acid substitutions are conservative substitutions.

[0139] In some embodiments, the amino acid substitutions are highly conservative substitutions.

[0140] In some embodiments, the polypeptide has a V that has 100% sequence identity to the amino acid sequence of SEQ ID NO: 19. L In some embodiments, the polypeptide comprises a V comprising the amino acid sequence of SEQ ID NO: 19. L Includes.

[0141] In some embodiments, the polypeptide has a V having at least about 70% sequence identity to any one or more amino acid sequences of SEQ ID NOs: 20-30. L The sequences identified as SEQ ID NOs: 20 to 30 are shown in Table 2 and are representative of human V LIn some embodiments, the polypeptide corresponds to a V domain having at least about 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one or more of the amino acid sequences of SEQ ID NOs: 20-30. L In some embodiments, the polypeptide comprises a V having at least about 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 20-30. L In some embodiments, the polypeptide comprises a V having at least about 90% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 20-30. L Includes.

[0142] In some embodiments, the polypeptide comprises a V that contains at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 20-30. L For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a V that contains about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 20-30. L Includes.

[0143] In some embodiments, the polypeptide has a V having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 20-30. L In some embodiments, the polypeptide comprises a V comprising the amino acid sequence of any one of SEQ ID NOs: 20-30. L Includes.

[0144] In some embodiments, the polypeptide has a V having at least about 70% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 20-22. L In some embodiments, the polypeptide comprises a V having at least about 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 20-22. L In some embodiments, the polypeptide comprises a V having at least about 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 20-22. L In some embodiments, the polypeptide comprises a V having at least about 90% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 20-22. L Includes.

[0145] In some embodiments, the polypeptide comprises a V that contains at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 20-22. L For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a V that contains about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 20-22. L Includes.

[0146] In some embodiments, the polypeptide has a V having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 20-22. L In some embodiments, the polypeptide comprises a V comprising the amino acid sequence of any one of SEQ ID NOs: 20-22. L Includes.

[0147] In some embodiments, the polypeptide is V H (e.g., rodent (e.g., mouse) V H , primate (e.g., human) V H Mammal V H ) and V L (e.g., rodent (e.g., mouse) V L , primate (e.g., human) V L Mammal V L In some embodiments, the polypeptide comprises a V that is humanized (e.g., at least about: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% humanized), includes human framework regions, or both. H and V L Includes.

[0148] In some embodiments, the polypeptide is a) V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 3 H ; b) V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 19 L , or Both a) and b) Includes.

[0149] In some embodiments, the polypeptide is a) V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 3 H and b) V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO: 19 L Includes.

[0150] In some embodiments, the polypeptide is a) V having at least about 55% (e.g., at least about 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity with the amino acid sequence of SEQ ID NO:3 H ; b) V having at least about 55% (e.g., at least about 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of SEQ ID NO: 19 L , or Both a) and b) Including, wherein the polypeptide is V of SEQ ID NO:3 H Amino acid sequence and V of SEQ ID NO: 19 L It does not contain all six CDRs of the antibody comprising the amino acid sequence.

[0151] In some embodiments, the polypeptide is a) V having at least about 55% (e.g., at least about 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity with the amino acid sequence of SEQ ID NO:3 H and b) V having at least about 55% (e.g., at least about 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of SEQ ID NO: 19 L Including, wherein the polypeptide is V of SEQ ID NO:3 H Amino acid sequence and V of SEQ ID NO: 19 L It does not contain all six CDRs of the antibody comprising the amino acid sequence.

[0152] In some embodiments, the polypeptide is a) V that contains at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative substitution) compared to the amino acid sequence of SEQ ID NO: 3 H ; b) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative substitution) compared to the amino acid sequence of SEQ ID NO: 19. L , or Both a) and b) Includes.

[0153] In some embodiments, the polypeptide is a) V that contains at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to the amino acid sequence of SEQ ID NO: 3 H and b) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to the amino acid sequence of SEQ ID NO: 19. L Includes.

[0154] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residue of SEQ ID NO: 3 H ; b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the LCDR1, LCDR2 and / or LCDR3 residue of SEQ ID NO: 19. L , or Both a) and b) Includes.

[0155] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residue of SEQ ID NO: 3 H and b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the LCDR1, LCDR2 and / or LCDR3 residue of SEQ ID NO: 19. L Includes.

[0156] In some embodiments, the polypeptide is a) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 3. H ; b) V includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 19. L , or Both a) and b) Includes.

[0157] In some embodiments, the polypeptide is a) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 3. H and b) V includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 19. L Includes.

[0158] In some embodiments, the polypeptide is a) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 4 to 17 H ; b) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 20 to 30 L , or Both a) and b) Includes.

[0159] In some embodiments, the polypeptide is a) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 4 to 17 H and b) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 20 to 30 L Includes.

[0160] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 4 to 17 H ; b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 20 to 30 L , or Both a) and b) Includes.

[0161] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 4 to 17 H and b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 20 to 30 L Includes.

[0162] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of HCDR1, HCDR2, and / or HCDR3 residues of any one or more of SEQ ID NOs: 4 to 17. H ; b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more LCDR1, LCDR2, and / or LCDR3 residues of SEQ ID NOs: 20 to 30. L , or Both a) and b) Includes.

[0163] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of HCDR1, HCDR2, and / or HCDR3 residues of any one or more of SEQ ID NOs: 4 to 17. H and b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more LCDR1, LCDR2, and / or LCDR3 residues of SEQ ID NOs: 20 to 30. L Includes.

[0164] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of any one or more of SEQ ID NOs: 4-17 H ; b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more non-CDR residues (e.g., within the framework regions) of SEQ ID NOs: 20-30. L , or Both a) and b) Includes.

[0165] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of any one or more of SEQ ID NOs: 4-17 Hand b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more non-CDR residues (e.g., within the framework regions) of SEQ ID NOs: 20-30. L Includes.

[0166] In some embodiments, the polypeptides disclosed herein are a) V comprising any one of the amino acid sequences of SEQ ID NOs: 4 to 17 H and b) V comprising any one of the amino acid sequences of SEQ ID NOs: 20 to 30 L Includes.

[0167] In some embodiments, the polypeptide is a) V comprising any one of the amino acid sequences of SEQ ID NOs: 4 to 17 H and b) V comprising the amino acid sequence of SEQ ID NO: 19 L Includes.

[0168] In some embodiments, the polypeptide is a) V comprising the amino acid sequence of SEQ ID NO: 3 H and b) V comprising any one of the amino acid sequences of SEQ ID NOs: 20 to 30 L Includes.

[0169] In some embodiments, the polypeptide is a) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 4 to 7 H ; b) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 20 to 22 L , or Both a) and b) Includes.

[0170] In some embodiments, the polypeptide is a) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 4 to 7 H and b) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with any one or more amino acid sequences of SEQ ID NOs: 20 to 22 L Includes.

[0171] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 4 to 7 H ; b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 20 to 22 L , or Both a) and b) Includes.

[0172] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 4 to 7 H and b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to any one or more amino acid sequences of SEQ ID NOs: 20 to 22 L Includes.

[0173] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of HCDR1, HCDR2, and / or HCDR3 residues of any one or more of SEQ ID NOs: 4 to 7. H ; b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more LCDR1, LCDR2, and / or LCDR3 residues of SEQ ID NOs: 20 to 22 L , or Both a) and b) Includes.

[0174] In some embodiments, the polypeptide is a) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of HCDR1, HCDR2, and / or HCDR3 residues of any one or more of SEQ ID NOs: 4 to 7. H and b) V containing at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more LCDR1, LCDR2, and / or LCDR3 residues of SEQ ID NOs: 20 to 22 L Includes.

[0175] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of any one or more of SEQ ID NOs: 4-7 H ; b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more non-CDR residues (e.g., within the framework regions) of SEQ ID NOs: 20-22. L , or Both a) and b) Includes.

[0176] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of any one or more of SEQ ID NOs: 4-7 H and b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of any one or more non-CDR residues (e.g., within the framework regions) of SEQ ID NOs: 20-22. L Includes.

[0177] In some embodiments, the polypeptides disclosed herein are a) V comprising any one of the amino acid sequences of SEQ ID NOs: 4 to 7 H and b) V comprising any one of the amino acid sequences of SEQ ID NOs: 20 to 22 L Includes.

[0178] In some embodiments, the polypeptide is a) V comprising any one of the amino acid sequences of SEQ ID NOs: 4 to 7 H and b) V comprising the amino acid sequence of SEQ ID NO: 19 L Includes.

[0179] In some embodiments, the polypeptide is a) V comprising the amino acid sequence of SEQ ID NO: 3 H and b) V comprising any one of the amino acid sequences of SEQ ID NOs: 20 to 22 L Includes.

[0180] In some embodiments, the polypeptide is a) a V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with the amino acid sequence of SEQ ID NO:5 H ; b) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with the amino acid sequence of SEQ ID NO: 21 L , or Both a) and b) Includes.

[0181] In some embodiments, the polypeptide is a) a V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with the amino acid sequence of SEQ ID NO:5 H and b) V having at least about 70% (e.g., at least about 75, 80, 85, 90, 95, 98, or 99%) sequence identity with the amino acid sequence of SEQ ID NO: 21 L Includes.

[0182] In some embodiments, the polypeptide is a) V that contains at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to the amino acid sequence of SEQ ID NO: 5 H ; b) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to the amino acid sequence of SEQ ID NO: 21. L , or Both a) and b) Includes.

[0183] In some embodiments, the polypeptide is a) V that contains at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to the amino acid sequence of SEQ ID NO: 5 H and b) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) compared to the amino acid sequence of SEQ ID NO: 21. L Includes.

[0184] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residue of SEQ ID NO: 5. H ; b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the LCDR1, LCDR2 and / or LCDR3 residue of SEQ ID NO: 21. L , or Both a) and b) Includes.

[0185] In some embodiments, the polypeptide is a) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residue of SEQ ID NO: 5. H and b) V comprising at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of the LCDR1, LCDR2 and / or LCDR3 residue of SEQ ID NO: 21. L Includes.

[0186] In some embodiments, the polypeptide is a) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 5. H ; b) V includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 21. L , or Both a) and b) Includes.

[0187] In some embodiments, the polypeptide is a) V that includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 5. H and b) V includes at least one amino acid substitution (e.g., at least one conservative substitution, such as a highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region) of SEQ ID NO: 21. L Includes.

[0188] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 4 H , and b) V comprising the amino acid sequence of SEQ ID NO: 20 L (AB-1) Includes.

[0189] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 5 H , and b) V comprising the amino acid sequence of SEQ ID NO: 21 L (AB-2) Includes.

[0190] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 6 H , and b) V comprising the amino acid sequence of SEQ ID NO: 20 L (AB-3) Includes.

[0191] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 7 H , and b) V comprising the amino acid sequence of SEQ ID NO: 22 L (AB-4) Includes.

[0192] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 8 H , and b) V comprising the amino acid sequence of SEQ ID NO: 23 L (AB-5) Includes.

[0193] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 9 H , and b) V comprising the amino acid sequence of SEQ ID NO: 24 L (AB-6) Includes.

[0194] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 10 H , and b) V comprising the amino acid sequence of SEQ ID NO: 25 L (AB-7) Includes.

[0195] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 11 H , and b) V comprising the amino acid sequence of SEQ ID NO: 20 L (AB-8) Includes.

[0196] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 12 H , and b) V comprising the amino acid sequence of SEQ ID NO: 20 L (AB-9) Includes.

[0197] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 13H , and b) V comprising the amino acid sequence of SEQ ID NO: 26 L (AB-10) Includes.

[0198] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 14 H , and b) V comprising the amino acid sequence of SEQ ID NO: 27 L (AB-11) Includes.

[0199] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 15 H , and b) V comprising the amino acid sequence of SEQ ID NO: 28 L (AB-12) Includes.

[0200] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 16 H , and b) V comprising the amino acid sequence of SEQ ID NO: 29 L (AB-13) Includes.

[0201] In some embodiments, the polypeptides disclosed herein are a) V comprising the amino acid sequence of SEQ ID NO: 17 H , and b) V comprising the amino acid sequence of SEQ ID NO: 30 L (AB-14) Includes.

[0202] Complementarity-Determining Regions (CDRs) The CDRs (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and / or LCDR3) may be CDRs defined by any art-recognized method for identifying CDR residues of an antibody (e.g., a CDR as defined by Kabat, a CDR as defined by Chothia, or a CDR as defined by IMGT), as further described herein.

[0203] In some embodiments, the polypeptide (e.g., antibody or antigen-binding fragment thereof) comprises V of SEQ ID NO:3. H Amino acid sequence and V of SEQ ID NO: 19 L In some embodiments, the polypeptide does not comprise all six CDRs of an antibody comprising the amino acid sequences SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:48, SEQ ID NO:59, SEQ ID NO:67, and SEQ ID NO:69. See Table 3 for SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:48, SEQ ID NO:59, SEQ ID NO:67, and SEQ ID NO:69. In some embodiments, the polypeptide does not comprise all four of the sequences SEQ ID NO:33, SEQ ID NO:48, SEQ ID NO:59, and SEQ ID NO:69.

[0204] In some embodiments, the polypeptide comprises V of SEQ ID NO:3. H Amino acid sequence and V of SEQ ID NO: 19 L The antibody comprises fewer than six (e.g., one, two, three, four, or five) CDRs comprising the amino acid sequences. In some embodiments, the polypeptide comprises one, two, three, four, or five, but not all six, sequences selected from SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:48, SEQ ID NO:59, SEQ ID NO:67, and SEQ ID NO:69. In some embodiments, the polypeptide comprises one, two, or three, but not all four, sequences selected from SEQ ID NO:33, SEQ ID NO:48, SEQ ID NO:59, and SEQ ID NO:69.

[0205] In some embodiments, the polypeptide comprises all six CDRs of a specific polypeptide disclosed herein, hi some embodiments, the polypeptide comprises fewer than six (e.g., 1, 2, 3, 4, or 5) of the CDRs of a specific polypeptide disclosed herein.

[0206] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to HCDR1, HCDR2, and HCDR3, respectively. H amino acid sequence; b) one or more V of SEQ ID NOs: 20 to 30 L V comprising LCDR1, LCDR2 and LCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to LCDR1, LCDR2 and LCDR3, respectively. L an amino acid sequence, or Both a) and b) Includes.

[0207] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to HCDR1, HCDR2, and HCDR3, respectively. H the amino acid sequence; and b) one or more V of SEQ ID NOs: 20 to 30 L V comprising LCDR1, LCDR2 and LCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to LCDR1, LCDR2 and LCDR3, respectively. L Amino acid sequence Includes.

[0208] See Tables 2-3 and Figures 2A-3B for non-limiting examples of SEQ ID NOs: 4-17 and 20-30 and corresponding HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences.

[0209] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 that substantially retain one or more functional properties of HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) one or more V of SEQ ID NOs: 20 to 30 L and V comprising LCDR1, LCDR2, and LCDR3, which substantially retain one or more functional properties of the amino acid sequences of LCDR1, LCDR2, and LCDR3, respectively. L an amino acid sequence, or Both a) and b) Includes.

[0210] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 that substantially retain one or more functional properties of HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) any one or more V of SEQ ID NOs: 20 to 30 L and V comprising LCDR1, LCDR2, and LCDR3, which substantially retain one or more functional properties of the amino acid sequences of LCDR1, LCDR2, and LCDR3, respectively. L Amino acid sequence Includes.

[0211] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 HV comprising HCDR1, HCDR2, and HCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) one or more V of SEQ ID NOs: 20 to 30 L V comprising LCDR1, LCDR2, and LCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0212] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) one or more V of SEQ ID NOs: 20 to 30 L V comprising LCDR1, LCDR2, and LCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L Amino acid sequence Includes.

[0213] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 that contain up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) one or more V of SEQ ID NOs: 20 to 30 LV comprising LCDR1, LCDR2, and LCDR3 that comprise up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0214] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 that contain up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) one or more V of SEQ ID NOs: 20 to 30 L V comprising LCDR1, LCDR2, and LCDR3 that comprise up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L Amino acid sequence Includes.

[0215] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 that have 100% sequence identity with HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence H an amino acid sequence; or b) one or more V of SEQ ID NOs: 20 to 30 L V comprising LCDR1, LCDR2, and LCDR3 having 100% sequence identity with LCDR1, LCDR2, and LCDR3, respectively, in amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0216] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 17 H V comprising HCDR1, HCDR2, and HCDR3 that have 100% sequence identity with HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence H the amino acid sequence; and b) any one or more V of SEQ ID NOs: 20 to 30 L V comprising LCDR1, LCDR2, and LCDR3 having 100% sequence identity with LCDR1, LCDR2, and LCDR3, respectively, in amino acid sequence. L Amino acid sequence Includes.

[0217] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14) V to be selected from H / V L The antibody comprises a combination of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3.

[0218] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 31; b) HCDR2 comprising at least one amino acid substitution compared to at least one amino acid sequence set forth in SEQ ID NOs: 33 to 46 (e.g., at least one amino acid sequence set forth in SEQ ID NOs: 34 to 46); c) an HCDR3 comprising at least one amino acid substitution compared to at least one amino acid sequence set forth in SEQ ID NOs: 48 to 57 (e.g., at least one amino acid sequence set forth in SEQ ID NOs: 49 to 57); d) LCDR1 comprising at least one amino acid substitution compared to at least one amino acid sequence set forth in SEQ ID NOs: 59 to 66 (e.g., at least one amino acid sequence set forth in SEQ ID NOs: 60 to 66); e) LCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 67; f) LCDR3 comprising at least one amino acid substitution compared to at least one amino acid sequence set forth in SEQ ID NOs: 69 to 80 (e.g., at least one amino acid sequence set forth in SEQ ID NOs: 70 to 80); or any combination of the foregoing Includes.

[0219] In some embodiments, the polypeptide is a) V shown in any one of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to HCDR1, HCDR2, and HCDR3, respectively. H amino acid sequence; b) V shown in any one of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2 and LCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to LCDR1, LCDR2 and LCDR3, respectively. L an amino acid sequence, or Both a) and b) Includes.

[0220] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to HCDR1, HCDR2, and HCDR3, respectively. H amino acid sequence; b) any one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2 and LCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to LCDR1, LCDR2 and LCDR3, respectively. L an amino acid sequence, or Both a) and b) Includes.

[0221] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to HCDR1, HCDR2, and HCDR3, respectively. H the amino acid sequence; and b) one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2 and LCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to LCDR1, LCDR2 and LCDR3, respectively. L Amino acid sequence Includes.

[0222] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 that substantially retain one or more functional properties of HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. Han amino acid sequence; or b) any one or more V of SEQ ID NOs: 20 to 22 L and V comprising LCDR1, LCDR2, and LCDR3, which substantially retain one or more functional properties of the amino acid sequences of LCDR1, LCDR2, and LCDR3, respectively. L an amino acid sequence, or Both a) and b) Includes.

[0223] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 that substantially retain one or more functional properties of HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) any one or more V of SEQ ID NOs: 20 to 22 L and V comprising LCDR1, LCDR2, and LCDR3, which substantially retain one or more functional properties of the amino acid sequences of LCDR1, LCDR2, and LCDR3, respectively. L Amino acid sequence Includes.

[0224] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) any one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2, and LCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0225] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) any one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2, and LCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L Amino acid sequence Includes.

[0226] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 that contain up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) any one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2, and LCDR3 that comprise up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0227] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 HV comprising HCDR1, HCDR2, and HCDR3 that contain up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2, and LCDR3 that comprise up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L Amino acid sequence Includes.

[0228] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 that have 100% sequence identity with HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence H an amino acid sequence; or b) one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2, and LCDR3 having 100% sequence identity with LCDR1, LCDR2, and LCDR3, respectively, in amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0229] In some embodiments, the polypeptide is a) one or more V of SEQ ID NOs: 4 to 7 H V comprising HCDR1, HCDR2, and HCDR3 that have 100% sequence identity with HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence H the amino acid sequence; and b) one or more V of SEQ ID NOs: 20 to 22 L V comprising LCDR1, LCDR2, and LCDR3 having 100% sequence identity with LCDR1, LCDR2, and LCDR3, respectively, in amino acid sequence.L Amino acid sequence Includes.

[0230] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4) V to be selected from H / V L The antibody comprises a combination of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3.

[0231] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 31; b) HCDR2 comprising at least one amino acid substitution compared to at least one amino acid sequence set forth in SEQ ID NOs: 33 to 37 (e.g., at least one amino acid sequence set forth in SEQ ID NOs: 34 to 37); c) an HCDR3 comprising at least one amino acid substitution compared to at least one amino acid sequence set forth in SEQ ID NOs: 48 to 52 (e.g., at least one amino acid sequence set forth in SEQ ID NOs: 49 to 52); d) LCDR1 comprising at least one amino acid substitution compared to at least one amino acid sequence shown in SEQ ID NO: 59 to 61 (e.g., at least one amino acid sequence shown in SEQ ID NO: 60 or SEQ ID NO: 61); e) LCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 67; f) LCDR3 comprising at least one amino acid substitution compared to at least one amino acid sequence set forth in SEQ ID NOs: 69 to 72 (e.g., at least one amino acid sequence set forth in SEQ ID NOs: 70 to 72); or any combination of the foregoing Includes.

[0232] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to HCDR1, HCDR2, and HCDR3, respectively. H amino acid sequence; b) V of SEQ ID NO: 21 L V comprising LCDR1, LCDR2 and LCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to LCDR1, LCDR2 and LCDR3, respectively. L an amino acid sequence, or Both a) and b) Includes.

[0233] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to HCDR1, HCDR2, and HCDR3, respectively. H the amino acid sequence; and b) V of SEQ ID NO: 21 L V comprising LCDR1, LCDR2 and LCDR3 whose amino acid sequences are substantially similar (e.g., have at least about 90% sequence identity) to LCDR1, LCDR2 and LCDR3, respectively. L Amino acid sequence Includes.

[0234] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 that substantially retain one or more functional properties of HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) V of SEQ ID NO: 21 Land V comprising LCDR1, LCDR2, and LCDR3, which substantially retain one or more functional properties of the amino acid sequences of LCDR1, LCDR2, and LCDR3, respectively. L an amino acid sequence, or Both a) and b) Includes.

[0235] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 that substantially retain one or more functional properties of HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) V of SEQ ID NO: 21 L and V comprising LCDR1, LCDR2, and LCDR3, which substantially retain one or more functional properties of the amino acid sequences of LCDR1, LCDR2, and LCDR3, respectively. L Amino acid sequence Includes.

[0236] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) V of SEQ ID NO: 21 L V comprising LCDR1, LCDR2, and LCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0237] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 HV comprising HCDR1, HCDR2, and HCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) V of SEQ ID NO: 21 L V comprising LCDR1, LCDR2, and LCDR3 that contain only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L Amino acid sequence Includes.

[0238] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 that contain up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H an amino acid sequence; or b) V of SEQ ID NO: 21 L V comprising LCDR1, LCDR2, and LCDR3 that comprise up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0239] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 that contain up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence. H the amino acid sequence; and b) V of SEQ ID NO: 21 LV comprising LCDR1, LCDR2, and LCDR3 that comprise up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions) relative to LCDR1, LCDR2, and LCDR3, respectively, of the amino acid sequence. L Amino acid sequence Includes.

[0240] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 that have 100% sequence identity with HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence H an amino acid sequence; or b) V of SEQ ID NO: 21 L V comprising LCDR1, LCDR2, and LCDR3 having 100% sequence identity with LCDR1, LCDR2, and LCDR3, respectively, in amino acid sequence. L an amino acid sequence, or Both a) and b) Includes.

[0241] In some embodiments, the polypeptide is a) V of SEQ ID NO: 5 H V comprising HCDR1, HCDR2, and HCDR3 that have 100% sequence identity with HCDR1, HCDR2, and HCDR3, respectively, of the amino acid sequence H the amino acid sequence; and b) V of SEQ ID NO: 21 L V comprising LCDR1, LCDR2, and LCDR3 having 100% sequence identity with LCDR1, LCDR2, and LCDR3, respectively, in amino acid sequence. L Amino acid sequence Includes.

[0242] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 4 H , and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 20 L(AB-1) Includes.

[0243] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 5 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 21 L (AB-2) Includes.

[0244] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 6 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 20 L (AB-3) Includes.

[0245] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 7 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 22 L (AB-4) Includes.

[0246] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 8 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 23 L (AB-5) Includes.

[0247] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 9 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 24 L (AB-6) Includes.

[0248] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 10 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 25 L (AB-7) Includes.

[0249] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 11 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 20 L (AB-8) Includes.

[0250] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 12 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 20 L (AB-9) Includes.

[0251] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 13 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 26 L (AB-10) Includes.

[0252] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 14 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 27 L (AB-11) Includes.

[0253] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 15 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 28 L (AB-12) Includes.

[0254] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 16 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 29 L (AB-13) Includes.

[0255] In some embodiments, the polypeptide is a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 17 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 30 L (AB-14) Includes.

[0256] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 34; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 49; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 70; or Any combination of the above Includes.

[0257] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 35; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 50; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 71; or Any combination of the above Includes.

[0258] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 36; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 51; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 70; or Any combination of the above Includes.

[0259] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 37; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 52; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 61; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 72, or Any combination of the above Includes.

[0260] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 38; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 53; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 59; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 73, or Any combination of the above Includes.

[0261] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 39; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 53; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 74, or Any combination of the above Includes.

[0262] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 40; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 49; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 62; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 75, or Any combination of the above Includes.

[0263] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 41; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 54; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 70; or Any combination of the above Includes.

[0264] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 41; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 51; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 70; or Any combination of the above Includes.

[0265] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 42; c) an HCDR3 comprising the amino acid sequence of SEQ ID NO: 53; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 63; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 76, or Any combination of the above Includes.

[0266] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 43; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 55; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 64; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 77, or Any combination of the above Includes.

[0267] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 44; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 56; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 65; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 78, or Any combination of the above Includes.

[0268] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 45; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 57; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 66; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 79, or Any combination of the above Includes.

[0269] In some embodiments, the polypeptide is a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31; b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 46; c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 48; d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 63; e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 67; f) an LCDR3 comprising the amino acid sequence of SEQ ID NO: 80; or Any combination of the above Includes.

[0270] Paratope The amino acid residues of the paratope contribute to the interaction of the antibody with its target protein epitope. The interaction can be hydrogen bonding, salt bridges, van der Waals interactions, electrostatic interactions, hydrophobic interactions, π-interaction effects, ionic bonds, and / or any combination thereof. The interaction can be direct or indirect, e.g., via a coordinating intermediate molecule such as an ion or water. The residues of the paratope, in some embodiments, include only residues that are part of the defined CDRs. In some embodiments, the residues of the paratope further include one or more residues that are not part of the defined CDRs (e.g., residues within the defined framework regions).

[0271] In some embodiments, when the polypeptide is bound to the target antigen, the paratope is less than about 5.0 angstroms from the epitope on the target antigen, e.g., less than about 4.5, 4.0, 3.5, 3.0, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, or 0.9 angstroms from the epitope, or about 0. 9 to 5.0, 0.9 to 4.8, 1.0 to 5, 1.0 to 4.5, 1.0 to 4.0, 1.0 to 3.5, 1.1 to 3.5, 1.1 to 3.0, 1.2 to 3.0, 1.2 to 2.5, 1.3 to 2.5, 1.3 to 2.4, 1.4 to 2.4, 1.4 to 2.3, 1.5 to 2.3, 1.5 to 2.2, 1.6 to 2.2, 1.6 to 2.1, 1.7 to 2.1, 1.7 to 2.0, or 1.8 to 2.0 angstroms. In some embodiments, when the polypeptide binds to a target antigen, fewer than all of the amino acid residues in the paratope that contribute to the paratope (e.g., about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% of the amino acid residues) are oriented less than about 5.0 angstroms from the epitope on the target antigen.

[0272] In some embodiments, a polypeptide (e.g., an antibody or antigen-binding fragment thereof) comprising a paratope disclosed herein comprises V H and V L In some embodiments, the paratope residues comprise the V H and V L is included in the range of

[0273] In some embodiments, the polypeptide comprises the V of SEQ ID NO:3 / SEQ ID NO:19. H / V L The paratopes of the antibodies include combinations of different paratopes.

[0274] In some embodiments, the polypeptide comprises the V of SEQ ID NO:3 / SEQ ID NO:19. H / V Land paratopes that differ by 1 to 3 (e.g., 1, 2, or 3) residue substitutions (e.g., conservative substitutions, such as highly conservative substitutions).

[0275] In some embodiments, the polypeptide comprises the V of SEQ ID NO:3 / SEQ ID NO:19. H / V L In some embodiments, the polypeptide comprises a paratope substantially similar (e.g., having at least about 90% sequence identity) to the paratope of an antibody comprising the combination of V of SEQ ID NO:3 / V of SEQ ID NO:19. H / V L and substantially retains one or more functional properties thereof.

[0276] In some embodiments, the polypeptide comprises the V of SEQ ID NO:3 / SEQ ID NO:19. H / V L The paratope of the antibody containing the combination of

[0277] In some embodiments, the polypeptide is a V selected from SEQ ID NO:4 / SEQ ID NO:20 (AB-1), SEQ ID NO:5 / SEQ ID NO:21 (AB-2), SEQ ID NO:6 / SEQ ID NO:20 (AB-3), SEQ ID NO:7 / SEQ ID NO:22 (AB-4), SEQ ID NO:8 / SEQ ID NO:23 (AB-5), SEQ ID NO:9 / SEQ ID NO:24 (AB-6), SEQ ID NO:10 / SEQ ID NO:25 (AB-7), SEQ ID NO:11 / SEQ ID NO:20 (AB-8), SEQ ID NO:12 / SEQ ID NO:20 (AB-9), SEQ ID NO:13 / SEQ ID NO:26 (AB-10), SEQ ID NO:14 / SEQ ID NO:27 (AB-11), SEQ ID NO:15 / SEQ ID NO:28 (AB-12), SEQ ID NO:16 / SEQ ID NO:29 (AB-13), or SEQ ID NO:17 / SEQ ID NO:30 (AB-14). H / V L The paratopes of the antibodies include combinations of different paratopes.

[0278] For SEQ ID NOs: 4 to 17, see Table 1, and for V shown in any one of SEQ ID NOs: 4 to 17, see Table 1. H See Figures 2A-2B for antibody paratope residues including sequences. See Table 2 for SEQ ID NOs: 20-30.

[0279] In some embodiments, the polypeptide is a V selected from SEQ ID NO:4 / SEQ ID NO:20 (AB-1), SEQ ID NO:5 / SEQ ID NO:21 (AB-2), SEQ ID NO:6 / SEQ ID NO:20 (AB-3), SEQ ID NO:7 / SEQ ID NO:22 (AB-4), SEQ ID NO:8 / SEQ ID NO:23 (AB-5), SEQ ID NO:9 / SEQ ID NO:24 (AB-6), SEQ ID NO:10 / SEQ ID NO:25 (AB-7), SEQ ID NO:11 / SEQ ID NO:20 (AB-8), SEQ ID NO:12 / SEQ ID NO:20 (AB-9), SEQ ID NO:13 / SEQ ID NO:26 (AB-10), SEQ ID NO:14 / SEQ ID NO:27 (AB-11), SEQ ID NO:15 / SEQ ID NO:28 (AB-12), SEQ ID NO:16 / SEQ ID NO:29 (AB-13), or SEQ ID NO:17 / SEQ ID NO:30 (AB-14). H / V L and a paratope that differs from the paratope of an antibody comprising a combination of the above by substitutions of 1 to 3 (e.g., 1, 2, or 3) residues (e.g., conservative substitutions, such as highly conservative substitutions).

[0280] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of these V to be selected from H / V L The antibody comprises a paratope that is substantially similar (e.g., has at least about 90% sequence identity) to the paratope of the antibody comprising the combination of:

[0281] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of these V to be selected from H / V L and substantially retains one or more functional properties thereof.

[0282] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of these V to be selected from H / V L The paratope includes a paratope that includes only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) compared to the paratope of an antibody that includes a combination of:

[0283] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of these V to be selected from H / V L and a paratope comprising up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 conservative substitutions (e.g., up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 highly conservative substitutions) compared to the paratope of an antibody comprising a combination of:

[0284] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of these V to be selected from H / V L and a paratope having 100% sequence identity with the paratope of an antibody comprising a combination of:

[0285] In some embodiments, the polypeptide is a V selected from SEQ ID NO:4 / SEQ ID NO:20 (AB-1), SEQ ID NO:5 / SEQ ID NO:21 (AB-2), SEQ ID NO:6 / SEQ ID NO:20 (AB-3), or SEQ ID NO:7 / SEQ ID NO:22 (AB-4). H / V L The paratopes of the antibodies include combinations of different paratopes.

[0286] In some embodiments, the polypeptide is a V selected from SEQ ID NO:4 / SEQ ID NO:20 (AB-1), SEQ ID NO:5 / SEQ ID NO:21 (AB-2), SEQ ID NO:6 / SEQ ID NO:20 (AB-3), or SEQ ID NO:7 / SEQ ID NO:22 (AB-4). H / V L and paratopes that differ by 1 to 3 (e.g., 1, 2, or 3) residue substitutions (e.g., conservative substitutions, such as highly conservative substitutions).

[0287] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), or A combination of these V to be selected from H / V L The antibody comprises a paratope that is substantially similar (e.g., has at least about 90% sequence identity) to the paratope of the antibody comprising the combination of:

[0288] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), or A combination of these V to be selected from H / V L and substantially retains one or more functional properties thereof.

[0289] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), or A combination of these V to be selected from H / V L The paratope includes a paratope that includes only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) compared to the paratope of an antibody that includes a combination of:

[0290] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), or A combination of these V to be selected from H / V L and a paratope comprising up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 conservative substitutions (e.g., up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 highly conservative substitutions) compared to the paratope of an antibody comprising a combination of:

[0291] In some embodiments, the polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), or A combination of these V to be selected from H / V L and a paratope having 100% sequence identity with the paratope of an antibody comprising a combination of:

[0292] In some embodiments, the polypeptide comprises a paratope comprising amino acid residues corresponding to each of M2, G26, F27, T28, R30, T31, Y32, Y53, R98, P100, Q101, W102, E103, and E107 of SEQ ID NO: 2, or a subset thereof. In some embodiments, the polypeptide comprises a paratope comprising amino acid residues corresponding to each of T28, T31, Y32, Y53, R98, W102, and E107 of SEQ ID NO: 2.

[0293] Consensus sequence In some embodiments, the polypeptide (e.g., an antibody or antigen-binding fragment thereof) has the amino acid sequence of SEQ ID NO:2, X1 is not I; X2 is not W; X3 is not D; X4 is not S; X5 is not N; X6 is not K; X7 is not A; X8 is not A; X9 is not L; X 10 is not V; X 11 is not H; X 12 is not A; X 13 is not F; or X 14 is not I, or V, including any combination of the above H Includes.

[0294] The sequence identified as SEQ ID NO: 2 is shown in Table 1 and corresponds to the consensus V of SEQ ID NOs: 3-17 herein. H It is an array.

[0295] In some embodiments, X1 is I or V; X2 is W or S; X3 is D or S; X4 is S, T, Y or A; X5 is N, Y, D or T; X6 is K, T or I; X7 is A, S or T; X8 is A or S; X9 is L, D, E or S; X 10 is V or I; X 11 is H, F or Y; X 12 is A or S; X 13 is F, M or L; X 14 is I or V, Or any combination of the above.

[0296] In some embodiments, X1 is V; X2 is S; X3 is S; X4 is T, Y or A; X5 is Y, D or T; X6 is T or I; X7 is S or T; X8 is S; X9 is D, E or S; X 10 is I; X 11 is F or Y; X 12 is S; X 13 is M or L; X 14 is V, Or any combination of the foregoing.

[0297] In some embodiments, X1 is not I. In some embodiments, X1 is I or V. In some embodiments, X1 is V. In some embodiments, X1 is I.

[0298] In some embodiments, X2 is not W. In some embodiments, X2 is W or S. In some embodiments, X2 is S. In some embodiments, X2 is W.

[0299] In some embodiments, X3 is not D. In some embodiments, X3 is D or S. In some embodiments, X3 is S. In some embodiments, X3 is D.

[0300] In some embodiments, X4 is not S. In some embodiments, X4 is S, T, Y, or A. In some embodiments, X4 is T, Y, or A. In some embodiments, X4 is T. In some embodiments, X4 is Y. In some embodiments, X4 is A. In some embodiments, X4 is S.

[0301] In some embodiments, X5 is not N. In some embodiments, X5 is N, Y, D, or T. In some embodiments, X5 is Y, D, or T. In some embodiments, X5 is Y. In some embodiments, X5 is D. In some embodiments, X5 is T. In some embodiments, X5 is N.

[0302] In some embodiments, X6 is not K. In some embodiments, X6 is K, T, or I. In some embodiments, X6 is T or I. In some embodiments, X6 is T. In some embodiments, X6 is I. In some embodiments, X6 is K.

[0303] In some embodiments, X7 is not A. In some embodiments, X7 is A, S, or T. In some embodiments, X7 is S or T. In some embodiments, X7 is S. In some embodiments, X7 is T. In some embodiments, X7 is A.

[0304] In some embodiments, X8 is not A. In some embodiments, X8 is A or S. In some embodiments, X8 is S. In some embodiments, X8 is A.

[0305] In some embodiments, X9 is not L. In some embodiments, X9 is L, D, E, or S. In some embodiments, X9 is D, E, or S. In some embodiments, X9 is D. In some embodiments, X9 is E. In some embodiments, X9 is S. In some embodiments, X9 is L.

[0306] In some embodiments, X 10 is not V. In some embodiments, X 10 is V or I. In some embodiments, X 10 is I. In some embodiments, X 10 is V.

[0307] In some embodiments, X 11 is not H. In some embodiments, X 11 is H, F or Y. In some embodiments, X 11 is F or Y. In some embodiments, X 11 is F. In some embodiments, X 11 is Y. In some embodiments, X 11 is H.

[0308] In some embodiments, X 12 is not A. In some embodiments, X 12 is A or S. In some embodiments, X 12 is S. In some embodiments, X12 is A.

[0309] In some embodiments, X 13 is not F. In some embodiments, X 13 is F, M, or L. In some embodiments, X 13 is M or L. In some embodiments, X 13 is M. In some embodiments, X 13 is L. In some embodiments, X 13 is F.

[0310] In some embodiments, X 14 is not I. In some embodiments, X 14 is I or V. In some embodiments, X 14 is V. In some embodiments, X 14 is I.

[0311] In some embodiments, the polypeptide has the amino acid sequence of SEQ ID NO: 18, wherein: X 15 is not N; X 16 is not L; X 17 is not S; X 18 is not K; X 19 is not S; X 20 is not V; X 21 is not W; X 22 is not D; X 23 is not S; X 24 is not S; X 25 is not S; X 26 is not D; or X 27 is not H, or any combination of the above) Contains V L Includes.

[0312] The sequence identified as SEQ ID NO: 18 is shown in Table 2 and corresponds to the consensus V of SEQ ID NOs: 19-30 herein. L It is an array.

[0313] In some embodiments, X 15 is N or Y; X 16 is L or I; X 17 is S or R; X 18 is K, F or Y; X 19 is S or N; X 20 is V or I; X 21 is W or Y; X 22 is D, V or S; X 23 is S, M or E; X 24 is S, A or T; X 25 is S, D or E; X 26 is D, S, R or F; X 27 is H, L, K or E, Or any combination of the above.

[0314] In some embodiments, X 15 is Y; X 16 is I; X 17 is R; X 18 is F or Y; X 19 is N; X 20 is I; X 21 is Y; X 22 is V or S; X 23 is M or E; X 24 is A or T; X 25 is D or E; X 26 is S, R or F; X 27 is L, K or E, Or any combination of the above.

[0315] In some embodiments, X 15 is not N. In some embodiments, X 15 is N or Y. In some embodiments, X 15 is Y. In some embodiments, X 15 is N.

[0316] In some embodiments, X 16 is not L. In some embodiments, X 16 is L or I. In some embodiments, X 16 is I. In some embodiments, X 16 is L.

[0317] In some embodiments, X 17 is not S. In some embodiments, X 17 is S or R. In some embodiments, X 17 is R. In some embodiments, X 17 is S.

[0318] In some embodiments, X 18 is not K. In some embodiments, X 18 is K, F, or Y. In some embodiments, X 18 is F or Y. In some embodiments, X 18 is F. In some embodiments, X 18 is Y. In some embodiments, X 18 is K.

[0319] In some embodiments, X19 is not S. In some embodiments, X 19 is S or N. In some embodiments, X 19 is N. In some embodiments, X 19 is S.

[0320] In some embodiments, X 20 is not V. In some embodiments, X 20 is V or I. In some embodiments, X 20 is I. In some embodiments, X 20 is V.

[0321] In some embodiments, X 21 is not W. In some embodiments, X 21 is W or Y. In some embodiments, X 21 is Y. In some embodiments, X 21 is W.

[0322] In some embodiments, X 22 is not D. In some embodiments, X 22 is D, V or S. In some embodiments, X 22 is V or S. In some embodiments, X 22 is V. In some embodiments, X 22 is S. In some embodiments, X 22 is D.

[0323] In some embodiments, X 23 is not S. In some embodiments, X 23 is S, M or E. In some embodiments, X 23 is M or E. In some embodiments, X 23 is M. In some embodiments, X 23 is E. In some embodiments, X 23 is S.

[0324] In some embodiments, X 24is not S. In some embodiments, X 24 is S, A, or T. In some embodiments, X 24 is A or T. In some embodiments, X 24 is A. In some embodiments, X 24 is T. In some embodiments, X 24 is S.

[0325] In some embodiments, X 25 is not S. In some embodiments, X 25 is S, D, or E. In some embodiments, X 25 is D or E. In some embodiments, X 25 is D. In some embodiments, X 25 is E. In some embodiments, X 25 is S.

[0326] In some embodiments, X 26 is not D. In some embodiments, X 26 is D, S, R or F. In some embodiments, X 26 is S, R or F. In some embodiments, X 26 is S. In some embodiments, X 26 is R. In some embodiments, X 26 is F. In some embodiments, X 26 is D.

[0327] In some embodiments, X 27 is not H. In some embodiments, X 27 is H, L, K, or E. In some embodiments, X 27 is L, K, or E. In some embodiments, X 27 is L. In some embodiments, X 27 is K. In some embodiments, X 27 is E. In some embodiments, X 27 is H.

[0328] In some embodiments, a) X1 is V, X2 is W, X3 is S, X4 is S, X5 is Y, X6 is T, X7 is S, X8 is S, X9 is D, and X 10 is I and X 11 is F and X 12 is A and X 13 is M and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; b) X1 is V, X2 is W, X3 is D, X4 is S, X5 is Y, X6 is T, X7 is A, X8 is S, X9 is E, and X 10 is I and X 11 is F and X 12 is A and X 13 is M and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is I and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; c) X1 is I, X2 is W, X3 is D, X4 is T, X5 is Y, X6 is T, X7 is A, X8 is S, X9 is S, and X 10 is I and X 11 is F and X 12 is A and X 13 is F and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; d) X1 is I, X2 is W, X3 is S, X4 is S, X5 is D, X6 is T, X7 is A, X8 is S, X9 is D, and X 10 is I and X 11 is F and X 12 is S and X 13 is M and X 14 is V and X 15 is Y and X 16 is I and X 17 is S and X 18 is F and X 19 is S and X 20 is I and X 21 is Y and X 22 is V and X 23 is S and X 24 is A and X 25 is S and X 26 is R, or X 27 is L, or a combination thereof; e) X1 is V, X2 is W, X3 is D, X4 is Y, X5 is D, X6 is T, X7 is S, X8 is S, X9 is D, and X10 is I and X 11 is F and X 12 is S and X 13 is M and X 14 is I and X 15 is N and X 16 is L and X 17 is S and X 18 is K and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is M and X 24 is S and X 25 is S and X 26 is D, or X 27 is L, or a combination thereof; f) X1 is V, X2 is W, X3 is D, X4 is S, X5 is D, X6 is I, X7 is S, X8 is S, X9 is D, and X 10 is I and X 11 is F and X 12 is S and X 13 is M and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is I and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is D, or X 27 is H, or a combination thereof; g) X1 is I, X2 is W, X3 is S, X4 is A, X5 is Y, X6 is I, X7 is S, X8 is S, X9 is D, and X 10 is I and X 11 is F and X 12 is A and X 13 is M and X14 is I and X 15 is N and X 16 is L and X 17 is S and X 18 is Y and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is M and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; h) X1 is I, X2 is W, X3 is D, X4 is S, X5 is Y, X6 is T, X7 is A, X8 is S, X9 is D, and X 10 is I and X 11 is F and X 12 is A and X 13 is F and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; i) X1 is I, X2 is W, X3 is D, X4 is S, X5 is Y, X6 is T, X7 is A, X8 is S, X9 is S, and X 10 is I and X 11 is F and X 12 is A and X 13 is F and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X18 is K and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; j) X1 is I, X2 is W, X3 is S, X4 is T, X5 is T, X6 is T, X7 is S, X8 is S, X9 is D, and X 10 is I and X 11 is F and X 12 is S and X 13 is M and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is N and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is D and X 26 is F, or X 27 is L, or a combination thereof; k) X1 is I, X2 is S, X3 is S, X4 is S, X5 is Y, X6 is I, X7 is T, X8 is S, X9 is E, and X 10 is V and X 11 is Y and X 12 is A and X 13 is L and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is F and X 19 is S and X 20 is V and X 21 is W and X22 is S and X 23 is S and X 24 is T and X 25 is S and X 26 is R, or X 27 is H, or a combination thereof; l) X1 is V, X2 is W, X3 is D, X4 is S, X5 is N, X6 is T, X7 is A, X8 is A, X9 is S, and X 10 is V and X 11 is H and X 12 is A and X 13 is F and X 14 is I and X 15 is N and X 16 is I and X 17 is R and X 18 is F and X 19 is S and X 20 is V and X 21 is W and X 22 is V and X 23 is S and X 24 is T and X 25 is S and X 26 is D, or X 27 is K, or a combination thereof; m) X1 is I, X2 is S, X3 is D, X4 is S, X5 is T, X6 is K, X7 is A, X8 is A, X9 is S, and X 10 is V and X 11 is Y and X 12 is A and X 13 is F and X 14 is I and X 15 is N and X 16 is L and X 17 is S and X 18 is F and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is E and X26 is D, or X 27 is L, or a combination thereof; or n) X1 is I, X2 is W, X3 is D, X4 is S, X5 is T, X6 is K, X7 is A, X8 is A, X9 is L, and X 10 is V and X 11 is H and X 12 is A and X 13 is F and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is N and X 20 is V and X 21 is W and X 22 is D and X 23 is E and X 24 is S and X 25 is S and X 26 is D, or X 27 is E, or a combination thereof.

[0329] In some embodiments, X1 is I or V; X2 is W; X3 is D or S; X4 is S or T; X5 is N, Y or D; X6 is K or T; X7 is A or S; X8 is A or S; X9 is L, D, E or S; X 10 is V or I; X 11 is H or F; X 12 is A or S; X 13 is F or M; or X 14 is I or V, or Any combination of the above.

[0330] In some embodiments, X1 is V; X2 is W; X3 is S; X4 is T; X5 is Y or D; X6 is T; X7 is S; X8 is S; X9 is D, E or S; X 10 is I; X 11 is F; X 12 is S; X 13 is M; X 14 is V, Or any combination of the above.

[0331] In some embodiments, X1 is not I. In some embodiments, X1 is I or V. In some embodiments, X1 is V. In some embodiments, X1 is I.

[0332] In some embodiments, X2 is W.

[0333] In some embodiments, X3 is not D. In some embodiments, X3 is D or S. In some embodiments, X3 is S. In some embodiments, X3 is D.

[0334] In some embodiments, X4 is not S. In some embodiments, X4 is S or T. In some embodiments, X4 is T. In some embodiments, X4 is S.

[0335] In some embodiments, X5 is not N. In some embodiments, X5 is N, Y, or D. In some embodiments, X5 is Y or D. In some embodiments, X5 is Y. In some embodiments, X5 is D. In some embodiments, X5 is N.

[0336] In some embodiments, X6 is not K. In some embodiments, X6 is K or T. In some embodiments, X6 is T. In some embodiments, X6 is K.

[0337] In some embodiments, X7 is not A. In some embodiments, X7 is A or S. In some embodiments, X7 is S. In some embodiments, X7 is A.

[0338] In some embodiments, X8 is not A. In some embodiments, X8 is A or S. In some embodiments, X8 is S. In some embodiments, X8 is A.

[0339] In some embodiments, X9 is not L. In some embodiments, X9 is L, D, E, or S. In some embodiments, X9 is D, E, or S. In some embodiments, X9 is D. In some embodiments, X9 is E. In some embodiments, X9 is S. In some embodiments, X9 is L.

[0340] In some embodiments, X 10 is not V. In some embodiments, X 10 is V or I. In some embodiments, X 10 is I. In some embodiments, X 10 is V.

[0341] In some embodiments, X 11 is not H. In some embodiments, X 11 is H or F. In some embodiments, X 11is F. In some embodiments, X 11 is H.

[0342] In some embodiments, X 12 is not A. In some embodiments, X 12 is A or S. In some embodiments, X 12 is S. In some embodiments, X 12 is A.

[0343] In some embodiments, X 13 is not F. In some embodiments, X 13 is F or M. In some embodiments, X 13 is M. In some embodiments, X 13 is F.

[0344] In some embodiments, X 14 is not I. In some embodiments, X 14 is I or V. In some embodiments, X 14 is V. In some embodiments, X 14 is I.

[0345] In some embodiments, X 15 is N or Y; X 16 is L or I; X 17 is S; X 18 is K or F; X 19 is S; X 20 is V or I; X 21 is W or Y; X 22 is D or V; X 23 is S; X 24 is S or A; X 25 is S; X26 is D, S or R; X 27 is H or L, Or any combination of the foregoing.

[0346] In some embodiments, X 15 is Y; X 16 is I; X 17 is S; X 18 is F; X 19 is S; X 20 is I; X 21 is Y; X 22 is V; X 23 is S; X 24 is A; X 25 is S; X 26 is S or R; X 27 is L, Or any combination of the foregoing.

[0347] In some embodiments, X 15 is not N. In some embodiments, X 15 is N or Y. In some embodiments, X 15 is Y. In some embodiments, X 15 is N.

[0348] In some embodiments, X 16 is not L. In some embodiments, X 16 is L or I. In some embodiments, X 16 is I. In some embodiments, X 16 is L.

[0349] In some embodiments, X 17is S.

[0350] In some embodiments, X 18 is not K. In some embodiments, X 18 is K or F. In some embodiments, X 18 is F. In some embodiments, X 18 is K.

[0351] In some embodiments, X 19 is S.

[0352] In some embodiments, X 20 is not V. In some embodiments, X 20 is V or I. In some embodiments, X 20 is I. In some embodiments, X 20 is V.

[0353] In some embodiments, X 21 is not W. In some embodiments, X 21 is W or Y. In some embodiments, X 21 is Y. In some embodiments, X 21 is W.

[0354] In some embodiments, X 22 is not D. In some embodiments, X 22 is D or V. In some embodiments, X 22 is V. In some embodiments, X 22 is D.

[0355] In some embodiments, X 23 is S.

[0356] In some embodiments, X 24 is not S. In some embodiments, X 24 is S or A. In some embodiments, X 24 is A. In some embodiments, X 24 is S.

[0357] In some embodiments, X 25 is S.

[0358] In some embodiments, X 26 is not D. In some embodiments, X 26 is D, S, or R. In some embodiments, X 26 is S or R. In some embodiments, X 26 is S. In some embodiments, X 26 is R. In some embodiments, X 26 is D.

[0359] In some embodiments, X 27 is not H. In some embodiments, X 27 is H or L. In some embodiments, X 27 is L. In some embodiments, X 27 is H.

[0360] In some embodiments, a) X1 is V, X2 is W, X3 is S, X4 is S, X5 is Y, X6 is T, X7 is S, X8 is S, X9 is D, and X 10 is I and X 11 is F and X 12 is A and X 13 is M and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; b) X1 is V, X2 is W, X3 is D, X4 is S, X5 is Y, X6 is T, X7 is A, X8 is S, X9 is E, and X 10 is I and X 11 is F and X 12 is A and X 13 is M and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is I and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; c) X1 is I, X2 is W, X3 is D, X4 is T, X5 is Y, X6 is T, X7 is A, X8 is S, X9 is S, and X 10 is I and X 11 is F and X 12 is A and X 13 is F and X 14 is I and X 15 is N and X 16 is I and X 17 is S and X 18 is K and X 19 is S and X 20 is V and X 21 is W and X 22 is D and X 23 is S and X 24 is S and X 25 is S and X 26 is S or X 27 is L, or a combination thereof; or d) X1 is I, X2 is W, X3 is S, X4 is S, X5 is D, X6 is T, X7 is A, X8 is S, X9 is D, and X10 is I and X 11 is F and X 12 is S and X 13 is M and X 14 is V and X 15 is Y and X 16 is I and X 17 is S and X 18 is F and X 19 is S and X 20 is I and X 21 is Y and X 22 is V and X 23 is S and X 24 is A and X 25 is S and X 26 is R, or X 27 is L, or a combination thereof.

[0361] Stationary Domain In some embodiments, the polypeptide (e.g., an antibody or antigen-binding fragment thereof) a) antibody heavy chain constant domain sequence; b) an antibody light chain constant domain sequence; or Both a) and b) Includes.

[0362] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence, hi some embodiments, the antibody heavy chain constant domain is selected from the group consisting of an IgA constant domain, an IgD constant domain, an IgE constant domain, an IgG constant domain, and an IgM constant domain.

[0363] In some embodiments, the IgG constant domain is an IgG1 constant domain (e.g., any one of SEQ ID NOs: 84-86), an IgG2 constant domain (e.g., SEQ ID NO: 87 or SEQ ID NO: 88), an IgG3 constant domain, or an IgG4 constant domain. In some embodiments, the IgG2 constant domain is an IgG2a, IgG2b, or IgG2c constant domain. In some embodiments, the IgA constant domain is an IgA1 constant domain or an IgA2 constant domain. In some embodiments, the antibody heavy chain constant domain is an IgG1 constant domain (e.g., IGHV1-5 or IGHV5-51). [ka] [ka] [ka] [ka] [ka]

[0364] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having at least about 60% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 84-88 (e.g., any one of SEQ ID NOs: 84-86). For example, the polypeptide may comprise an antibody heavy chain constant domain sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 84-88 (e.g., any one of SEQ ID NOs: 84-86). In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 84-88 (e.g., any one of SEQ ID NOs: 84-86).

[0365] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising at least one amino acid substitution compared to the amino acid sequence of any one of SEQ ID NOs: 84-88 (e.g., any one of SEQ ID NOs: 84-86). For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1 to 20, 1 to 19, 2 to 19, 2 to 18, 2 to 17, 3 to 17, 3 to 16, 4 to 16, 4 to 15, 5 to 15, 5 to 14, 6 to 14, 6 to 13, 7 to 13, 7 to 12, 8 to 12, 8 to 11, or 9 to 11. In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NOs: 84-88 (e.g., any one of SEQ ID NOs: 84-86).

[0366] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 84-88 (eg, any one of SEQ ID NOs: 84-86).

[0367] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 84. For example, the polypeptide can comprise an antibody heavy chain constant domain sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO:84. In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of SEQ ID NO:84.

[0368] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 84. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1 to 20, 1 to 19, 2 to 19, 2 to 18, 2 to 17, 3 to 17, 3 to 16, 4 to 16, 4 to 15, 5 to 15, 5 to 14, 6 to 14, 6 to 13, 7 to 13, 7 to 12, 8 to 12, 8 to 11, or 9 to 11. In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO:84.

[0369] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO: 84. Antibodies disclosed herein using the designation "AB-[]a" (e.g., AB-2a, see, e.g., Table 4) include those comprising the heavy chain constant domain amino acid sequence of SEQ ID NO: 84, i.e., M135, S137, and T139.

[0370] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence (e.g., an antibody or antigen-binding fragment thereof) having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 85. For example, the polypeptide can comprise an antibody heavy chain constant domain sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 85. In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of SEQ ID NO:85.

[0371] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 85. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1 to 20, 1 to 19, 2 to 19, 2 to 18, 2 to 17, 3 to 17, 3 to 16, 4 to 16, 4 to 15, 5 to 15, 5 to 14, 6 to 14, 6 to 13, 7 to 13, 7 to 12, 8 to 12, 8 to 11, or 9 to 11. In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO:85.

[0372] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO: 85. Antibodies disclosed herein using the designation "AB-[]b" (e.g., AB-2b, see, e.g., Table 4) include those that comprise the heavy chain constant domain amino acid sequence of SEQ ID NO: 85, i.e., Y135, T137, and E139).

[0373] In some embodiments, the polypeptide comprises an Fc polypeptide or an Fc domain (e.g., an IgG1 domain, an IgG2 domain, or an IgG4 domain). In some embodiments, the Fc domain comprises one or more mutations that, for example, reduce (e.g., inhibit, eliminate) an effector function of the Fc domain, increase the half-life of an antibody or antibody fragment comprising the Fc domain, or a combination thereof. See, e.g., Dumet et al., Insights into the IgG heavy chain engineering patent landscape as applied to IgG4 antibody development, Mabs. 11(8):1341-50 (2019) (especially Tables 1 and 2 therein) and WO 02060919, the contents of which are incorporated herein by reference in their entireties. In some embodiments, the Fc domain comprises an LS modification (i.e., an M428L / N434S substitution in the numbering determined by Kabat) or a YTE modification (i.e., an M252Y / S254T / T256E substitution as determined by Kabat numbering, which corresponds to M135Y / S137T / T139E when comparing SEQ ID NO: 84 to SEQ ID NO: 85). In some embodiments, the Fc domain comprises a YTE modification.

[0374] In some embodiments, the polypeptide (eg, an antibody or antigen-binding fragment thereof) comprises an antibody light chain constant domain sequence.

[0375] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence selected from the group consisting of a kappa constant domain (eg, SEQ ID NO: 89) and a lambda constant domain (eg, SEQ ID NO: 90). [ka] [ka]

[0376] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 89 or SEQ ID NO: 90. For example, the polypeptide can comprise an antibody light chain constant domain sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 89 or SEQ ID NO: 90. In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having at least about 70% or at least about 80% sequence identity to SEQ ID NO:89 or SEQ ID NO:90.

[0377] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO:89 or SEQ ID NO:90. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO:89 or SEQ ID NO:90. In some embodiments, at least one amino acid substitution is a conservative substitution. In some embodiments, at least one amino acid substitution is a highly conservative substitution.

[0378] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO:89 or SEQ ID NO:90.

[0379] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 90. For example, the polypeptide can comprise an antibody light chain constant domain sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90. In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having at least about 70% or at least about 80% sequence identity to SEQ ID NO:90.

[0380] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 90. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 90. In some embodiments, at least one amino acid substitution is a conservative substitution. In some embodiments, at least one amino acid substitution is a highly conservative substitution.

[0381] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO:90.

[0382] In some embodiments, the polypeptide is a) an antibody heavy chain constant domain sequence; and b) antibody light chain constant domain sequence Includes.

[0383] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain that is an IgG1 constant domain (eg, SEQ ID NO: 84 or SEQ ID NO: 85) and an antibody light chain constant domain that is a lambda constant domain (eg, SEQ ID NO: 90).

[0384] In some embodiments, the polypeptide is a) an antibody heavy chain constant domain comprising SEQ ID NO: 85; or b) an antibody light chain constant domain comprising SEQ ID NO: 90; Or both a) and b) Includes.

[0385] In some embodiments, the polypeptide is a) an antibody heavy chain constant domain comprising SEQ ID NO: 85; and b) an antibody light chain constant domain comprising SEQ ID NO: 90 Includes.

[0386] In some embodiments, the polypeptide is a) an antibody heavy chain constant domain comprising SEQ ID NO: 84; or b) an antibody light chain constant domain comprising SEQ ID NO: 90; Or both a) and b) Includes.

[0387] In some embodiments, the polypeptide is a) an antibody heavy chain constant domain comprising SEQ ID NO: 84; and b) an antibody light chain constant domain comprising SEQ ID NO: 90 Includes.

[0388] Antibodies and antigen-binding fragments In some embodiments, the polypeptide is an immunoglobulin molecule, such as an antibody (eg, a whole antibody, an intact antibody) or an antigen-binding fragment of an antibody.

[0389] In some embodiments, the polypeptide is an antibody.As used herein, the term "antibody" refers to an immunoglobulin molecule that has the ability to specifically bind to targets such as carbohydrates, polynucleotides, lipids, polypeptides, etc. through at least one antigen recognition site located in the variable domain of the immunoglobulin molecule.As used herein, the term "antibody" refers to a full-length antibody.

[0390] In some embodiments, the polypeptide is an antibody or multimer thereof (e.g., IgM) comprising two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. H and heavy chain constant domains (including domains CH1, hinge CH2 and CH3). L and a light chain constant domain (CL). H and V LThe regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) interspersed within framework regions (FRs). H and V L each comprises three CDRs and four FRs arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The antibody may be of any species, such as a murine antibody, a human antibody, or a humanized antibody.

[0391] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-118. For example, the polypeptide can comprise a heavy chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-118. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-118.

[0392] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 91-118. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 91-118.

[0393] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 91-118.

[0394] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-94 and 105-108. For example, the polypeptide may comprise a heavy chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-94 and 105-108. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 80% sequence identity to any one or more of the amino acid sequences of SEQ ID NOs: 91-94 and 105-108.

[0395] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 91-94 and 105-108. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 91-94 and 105-108.

[0396] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 91-94 and 105-108.

[0397] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-94. For example, the polypeptide can comprise a heavy chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-94. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 91-94.

[0398] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 91-94. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 91-94.

[0399] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 91-94.

[0400] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 92. For example, a polypeptide can comprise a heavy chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 92. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of SEQ ID NO:92.

[0401] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 92. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 92.

[0402] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO:92.

[0403] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 105-108. For example, the polypeptide can comprise a heavy chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 105-108. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 105-108.

[0404] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 105-108. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 105-108.

[0405] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 105-108.

[0406] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 106. For example, the polypeptide can comprise a heavy chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 106. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of SEQ ID NO:106.

[0407] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 106. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 106.

[0408] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO:106.

[0409] In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 119-129. For example, the polypeptide can comprise a light chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 119-129. In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 119-129.

[0410] In some embodiments, the polypeptide comprises a light chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 119-129. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a light chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 119-129.

[0411] In some embodiments, the polypeptide comprises a light chain amino acid sequence having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 119-129.

[0412] In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 119-121. For example, the polypeptide can comprise a light chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 119-121. In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 80% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs: 119-121.

[0413] In some embodiments, the polypeptide comprises a light chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of any one or more of SEQ ID NOs: 119-121. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a light chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 119-121.

[0414] In some embodiments, the polypeptide comprises a light chain amino acid sequence having 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 119-121.

[0415] In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 120. For example, the polypeptide can comprise a light chain amino acid sequence having at least about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 120. In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 70% or at least about 80% sequence identity to the amino acid sequence of SEQ ID NO:120.

[0416] In some embodiments, the polypeptide comprises a light chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 120. For example, the number of amino acid substitutions can be at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, the polypeptide comprises a light chain amino acid sequence comprising about 1-10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 120.

[0417] In some embodiments, the polypeptide comprises a light chain amino acid sequence having 100% sequence identity to the amino acid sequence of SEQ ID NO:120.

[0418] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to any one or more of the amino acid sequences of SEQ ID NOs: 91 to 118; or b) a light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences of SEQ ID NOs: 119 to 129; or Both a) and b) Includes.

[0419] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to any one or more amino acid sequences of SEQ ID NOs: 91 to 118; and b) a light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences of SEQ ID NOs: 119 to 129 Includes.

[0420] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 91 to 118; or b) a light chain amino acid sequence comprising at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 129; or Both a) and b) Includes.

[0421] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 91 to 118; and b) a light chain amino acid sequence containing at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 129; Includes.

[0422] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 91 to 118; or b) a light chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 129; or Both a) and b) Includes.

[0423] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequence of any one or more of SEQ ID NOs: 91 to 118; and b) a light chain amino acid sequence comprising approximately 1 to 10 amino acid substitutions compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 129; Includes.

[0424] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 91 to 118; or b) a light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 119 to 129, or Both a) and b) Includes.

[0425] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 91 to 118; and b) A light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 119 to 129 Includes.

[0426] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to any one or more of the amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; or b) a light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0427] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to any one or more of the amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; and b) a light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences of SEQ ID NOs: 119 to 121 Includes.

[0428] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to any one or more of the amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; or b) a light chain amino acid sequence comprising at least one amino acid substitution compared to any one or more of the amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0429] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to any one or more of the amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; and b) a light chain amino acid sequence containing at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 121; Includes.

[0430] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising approximately 1 to 10 amino acid substitutions compared to any one or more of the amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; or b) a light chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0431] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising approximately 1 to 10 amino acid substitutions compared to any one or more amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; and b) a light chain amino acid sequence comprising approximately 1 to 10 amino acid substitutions compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 121; Includes.

[0432] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; or b) a light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0433] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 91 to 94 and 105 to 108; and b) A light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 119 to 121 Includes.

[0434] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to any one or more amino acid sequences of SEQ ID NOs: 105 to 108; or b) a light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0435] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to any one or more amino acid sequences of SEQ ID NOs: 105 to 108; and b) a light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences of SEQ ID NOs: 119 to 121 Includes.

[0436] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 105 to 108; or b) a light chain amino acid sequence comprising at least one amino acid substitution compared to any one or more of the amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0437] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 105 to 108; and b) a light chain amino acid sequence containing at least one amino acid substitution compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 121; Includes.

[0438] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequences of any one or more of SEQ ID NOs: 105 to 108; or b) a light chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0439] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequences of any one or more of SEQ ID NOs: 105 to 108; and b) a light chain amino acid sequence comprising approximately 1 to 10 amino acid substitutions compared to any one or more amino acid sequences of SEQ ID NOs: 119 to 121; Includes.

[0440] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 105 to 108; or b) a light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 119 to 121; or Both a) and b) Includes.

[0441] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 105 to 108; and b) A light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 119 to 121 Includes.

[0442] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 106; or b) a light chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 120; or Both a) and b) Includes.

[0443] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence having at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 106; and b) a light chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO: 120 Includes.

[0444] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 106; or b) a light chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 120; or Both a) and b) Includes.

[0445] In some embodiments, the polypeptide is a) a heavy chain amino acid sequence comprising at least one amino acid substitution compared to the amino acid sequence of SEQ ID NO: 106; and b) a light chain amino acid sequence comprising about 1 to 10 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 120; Includes.

[0446] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 105; and b) a light chain (AB-1b) comprising the amino acid sequence of SEQ ID NO: 119 Includes.

[0447] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106; and b) a light chain (AB-2b) comprising the amino acid sequence of SEQ ID NO: 120 Includes.

[0448] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 107; and b) a light chain (AB-3b) comprising the amino acid sequence of SEQ ID NO: 119 Includes.

[0449] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 108; and b) a light chain (AB-4b) comprising the amino acid sequence of SEQ ID NO: 121 Includes.

[0450] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 109; and b) a light chain (AB-5b) comprising the amino acid sequence of SEQ ID NO: 122 Includes.

[0451] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 110; and b) a light chain (AB-6b) comprising the amino acid sequence of SEQ ID NO: 123 Includes.

[0452] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 111; and b) a light chain (AB-7b) comprising the amino acid sequence of SEQ ID NO: 124 Includes.

[0453] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 112; and b) a light chain (AB-8b) comprising the amino acid sequence of SEQ ID NO: 119 Includes.

[0454] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 113; and b) a light chain (AB-9b) comprising the amino acid sequence of SEQ ID NO: 119 Includes.

[0455] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 114; and b) a light chain (AB-10b) comprising the amino acid sequence of SEQ ID NO: 125 Includes.

[0456] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 115; and b) a light chain (AB-11b) comprising the amino acid sequence of SEQ ID NO: 126 Includes.

[0457] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 116; and b) a light chain (AB-12b) comprising the amino acid sequence of SEQ ID NO: 127 Includes.

[0458] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 117; and b) a light chain (AB-13b) comprising the amino acid sequence of SEQ ID NO: 128 Includes.

[0459] In some embodiments, the polypeptide is a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 118; and b) a light chain (AB-14b) comprising the amino acid sequence of SEQ ID NO: 129 Includes.

[0460] In some embodiments, the polypeptide is a single domain antibody or an antigen-binding fragment thereof. As used herein, the term "single domain antibody (sdAb)" or "nanobody" refers to an immunoglobulin molecule that consists of a single monomeric variable antibody domain and has the ability to specifically bind to a target. Single domain antibodies can be of any species, including murine, human, or humanized single domain antibodies.

[0461] In some embodiments, V H Domain and V L A variety of single chain antibody designs can be formed by linking domains together with linkers (e.g., synthetic linkers), where V H / V L Domains are paired intramolecularly or V H and V L When the domains are expressed by separate chains, they pair intermolecularly to form a monovalent antigen-binding site.

[0462] In some embodiments, the polypeptide is a heavy chain antibody, or an antigen-binding fragment thereof, that comprises two or more heavy chains but lacks light chains. Non-limiting examples of heavy chain antibodies include camelid Vhh (also known as VHH or V H H) Antibodies include camelid antibodies, which are antibodies from mammals of the Camelidae family, which includes llamas, camels, and alpacas.

[0463] In some embodiments, the polypeptide is an antibody mimetic. The term "antibody mimetic" refers to a polypeptide that has the ability to mimic the antigen-binding ability of an antibody but is structurally different from a natural antibody structure. Non-limiting examples of antibody mimetics include adnectins, affibodies, affilins, affimers, affitins, alphabodies, anticalins, avimers, DARPins, fynomers, Kunitz domain peptides, monobodies, nanobodies, nanoCLAMPs, and versabodies.

[0464] In some embodiments, the polypeptide is an antigen-binding fragment of an antibody. The term "antigen-binding fragment" refers to a portion of an immunoglobulin molecule (e.g., an antibody) that retains the antigen-binding properties of a full-length reference antibody. Non-limiting examples of antigen-binding fragments include V H area, V L region, Fab fragment, F(ab')2 fragment, Fd fragment, Fv fragment, and one V H Domain or one V L Examples include domain antibodies (dAbs) consisting of domains. H and V L A variety of single chain antibody designs can be formed by linking domains together with synthetic linkers, where V H / V L Domains are paired intramolecularly or V H and V L When the domains are expressed as separate chains, they pair intermolecularly to form a monovalent antigen-binding site, such as a single-chain Fv (scFv) or diabody. In some embodiments, the polypeptides disclosed herein are antigen-binding fragments selected from Fab, F(ab')2, Fab', scFv, or Fv. In some embodiments, the polypeptide is an scFv.

[0465] In some embodiments, the polypeptide is an isolated polypeptide.

[0466] In some embodiments, a polypeptide (e.g., an isolated polypeptide) is recombinantly produced. In some embodiments, a polypeptide (e.g., an isolated polypeptide) is synthetically produced.

[0467] In some embodiments, the polypeptide is linked to a second polypeptide. The term "linked" means attached via covalent or non-covalent interactions. A suitable linking agent can be used for conjugation. Non-limiting examples include peptide linkers, compound linkers, and chemical cross-linkers. In some embodiments, the linker is a disulfide bond.

[0468] In some embodiments, the polypeptide is conjugated to a heterologous moiety. The term "conjugated" refers to being attached via covalent or non-covalent interactions. Conjugation can use any one or more suitable linking agents. Non-limiting examples include peptide linkers, compound linkers, and chemical cross-linkers.

[0469] In some embodiments, the heterologous moiety comprises a therapeutic agent, a diagnostic agent, or both. In some embodiments, the heterologous moiety is selected from polyethylene glycol (PEG), hexadecanoic acid, a hydrogel, a nanoparticle, a multimerization domain, and a carrier peptide.

[0470] In some embodiments, the nanoparticles are lipid nanoparticles. In some embodiments, the nanoparticles are polymeric nanoparticles. In some embodiments, the polymer is an amphiphilic polymer. In some embodiments, the polymer is a hydrophobic or hydrophilic polymer. Non-limiting examples of polymers include poly(lactic acid)-poly(ethylene glycol), poly(lactic acid-co-glycolic acid)-poly(ethylene glycol), poly(lactic acid-co-glycolic acid) (PLGA), poly(lactic acid-co-glycolic acid)-d-α-tocopherol polyethylene glycol succinate, poly(lactic acid-co-glycolic acid)-ethylene oxide fumarate, poly(glycolic acid)-poly(ethylene glycol), polycaprolactone-poly(ethylene glycol), or any salt thereof. In some embodiments, the polymeric nanoparticles comprise poly(lactic acid-co-glycolic acid) (PLGA).

[0471] In some embodiments, the carrier polypeptide is albumin or an Fc polypeptide.

[0472] In some embodiments, the polypeptide is a) TSLP protein has a K of 10 pM or less D binds (e.g., as measured by KinExA); b) binds to the AB loop region and the C-terminal region of helix D of the TSLP protein; c) binds to an epitope on the TSLP protein (e.g., full-length human TSLP); d) competes with a reference antibody for binding to the TSLP protein; e) reducing the binding of TSLP protein to TSLPR; f) modulate (e.g., decrease, inhibit, neutralize) TSLP-mediated biological activity, or This is the combination mentioned above.

[0473] In some embodiments, the polypeptide has a K for TSLP (e.g., human) of 10 pM or less. D(e.g., as measured by KinExA).

[0474] In some embodiments, the polypeptide binds to (eg, has binding affinity for, has binding specificity for) the AB loop region and the C-terminal region of helix D of TSLP.

[0475] In some embodiments, the polypeptide has neutralizing activity against TSLP (e.g., full-length human TSLP). In some embodiments, the polypeptide has inhibitory activity against TSLP-mediated signaling.

[0476] In some embodiments, the polypeptide binds to a TSLP protein (e.g., SEQ ID NO: 1) at about 10 pM, 8 pM, 5 pM, 4 pM, 3 pM, 2 pM, 1.8 pM, 1.5 pM, 1.2 pM, 1.0 pM, 0.9 pM, 0.8 pM, 0.7 pM, 0.6 pM, 0.5 pM, 0.4 pM, 0.3 pM, 0.2 pM, or 0.1 pM, or less; or a K of about 0.1 to 10 pM, 0.2 to 10 pM, 0.2 to 8 pM, 0.3 to 8 pM, 0.3 to 5 pM, 0.4 to 5 pM, 0.4 to 4 pM, 0.5 to 4 pM, 0.5 to 3 pM, 0.6 to 3 pM, 0.6 to 2 pM, 0.7 to 2 pM, 0.7 to 1.8 pM, 0.8 to 1.8 pM, 0.8 to 1.5 pM, 1.0 to 1.5 pM, or 1.2 to 1.5 pM D In some embodiments, the polypeptide binds to the TSLP protein (e.g., SEQ ID NO: 1) with a K of 2 pM or less. D Combine with.

[0477] In some embodiments, the polypeptide is at about 2.0 pM, 1.9 pM, 1.8 pM, 1.7 pM, 1.6 pM, 1.5 pM, 1.4 pM, 1.3 pM, 1.2 pM, 1.1 pM, 1.0 pM, 0.9 pM, 0.8 pM, 0.7 pM, 0.6 pM, 0.5 pM, 0.4 pM, 0.3 pM, 0.2 pM, or 0.1 pM, or less, relative to the TSLP protein (e.g., SEQ ID NO: 1); or about 0.1 to 2.0 pM , 0.2-2.0pM, 0.2-1.9pM, 0.3-1.9pM, 0.3-1.8pM, 0.4-1.8pM, 0.4-1.7pM, 0.5-1.7pM, 0.5-1.6pM, 0.6-1.6pM, 0.6-1.5pM, 0.7-1.5pM, 0.7-1.4pM, 0.8-1.4pM, 0.8-1.3pM, 0.9-1.3pM, 1.0-1.3pM, 1.0-1.2pM or 1.1-1.2pM D In some embodiments, the polypeptide binds to TSLP with a K of about 1 pM or less. D Combine with.

[0478] In some embodiments, the polypeptide binds to the AB loop region and the C-terminal region of helix D of the TSLP protein.

[0479] In some embodiments, the polypeptide binds to an epitope of a TSLP protein (eg, full-length human TSLP).

[0480] In some embodiments, the polypeptide competes with a reference antibody for binding to a TSLP protein (e.g., SEQ ID NO: 1). Techniques and assays for assessing competition between antibodies are known in the art.

[0481] In some embodiments, the polypeptide reduces binding of a TSLP protein (e.g., a full-length TSLP protein such as SEQ ID NO: 1) to its receptor TSLPR (alone or in a heterodimeric complex with IL-7Rα). In some embodiments, the polypeptide reduces binding of a TSLP protein to TSLPR by about 500 pM or less, e.g., about 450 pM, 400 pM, 350 pM, 300 pM, 250 pM, 200 pM, 150 pM, 100 pM, 50 pM, 20 pM, 10 pM, 5 pM, 2 pM, 1 pM, 0.5 pM, 0.2 pM, or 0.1 pM or less; or about IC of 0.1-500pM, 0.1-450pM, 0.2-450pM, 0.2-400pM, 0.5-400pM, 0.5-350pM, 1-350pM, 1-300pM, 2-300pM, 2-250pM, 5-250pM, 5-200pM, 10-200pM, 10-150pM, 20-150pM, 20-100pM or 50-100pM 50 Decreases by.

[0482] In some embodiments, the polypeptide reduces binding of a TSLP protein (e.g., a full-length TSLP protein such as SEQ ID NO: 1) to TSLPR by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the polypeptide reduces binding of a TSLP protein (e.g., a full-length TSLP protein such as SEQ ID NO: 1) to TSLPR by at least about 30%.

[0483] In some embodiments, the polypeptide modulates (eg, reduces, inhibits, neutralizes) a TSLP-mediated biological activity.

[0484] Non-limiting examples of TSLP-mediated activities include TSLPR binding, TSLP-induced signal transduction and activator of transcription 5 (STAT5) signaling, TSLP-induced STAT3 signaling, TSLP-induced nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) signaling, TSLP-induced phosphoinositide-3-kinase (PI3K) signaling, TSLP-induced mitogen-activated protein kinase (MAPK) signaling, activation of one or more genes encoding interleukin-4 (IL-4), IL-5, IL-9, IL-13, and mitogen-activated protein kinase kinase 2 (MEK2), and dendritic cell activation. In some embodiments, TSLP-mediated activities involve one or more innate immune mechanisms and / or one or more adaptive immune mechanisms.

[0485] TSLP-mediated activity can be determined, for example, by measuring STAT5 reporter activity, dendritic cell activation, proliferation of cells expressing TSLPR, and CCL17 production (see, e.g., Verstraete et al., Structure and antagonism of the receptor complex mediated by human TSLP in allergy and asthma, Nat Commun. 8:14937 (2017), the contents of which are incorporated herein by reference).

[0486] In some embodiments, the polypeptide neutralizes an activity mediated by a TSLP protein (eg, a full-length TSLP protein such as SEQ ID NO:1).

[0487] In some embodiments, the polypeptides have a TSLP-mediated activity of about 500 pM or less, e.g., about 450 pM, 400 pM, 350 pM, 300 pM, 250 pM, 200 pM, 150 pM, 100 pM, 50 pM, 20 pM, 10 pM, 5 pM, 2 pM, 1 pM, 0.5 pM, 0.2 pM, or 0.1 pM, or less; or about 0.1 to IC of 500pM, 0.1-450pM, 0.2-450pM, 0.2-400pM, 0.5-400pM, 0.5-350pM, 1-350pM, 1-300pM, 2-300pM, 2-250pM, 5-250pM, 5-200pM, 10-200pM, 10-150pM, 20-150pM, 20-100pM or 50-100pM 50 Neutralize with.

[0488] In some embodiments, the polypeptide reduces TSLP-mediated activity by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the polypeptide reduces TSLP-mediated activity by at least about 30%.

[0489] In some embodiments, the polypeptide reduces a T2 inflammatory response mediated by one or more innate immune mechanisms of action of TSLP.

[0490] Non-limiting examples of T2 inflammatory responses include activation of type 2 innate lymphoid cells (ILC2s); activation, migration, and / or local differentiation of eosinophil progenitor cells (EoPs) (e.g., by upregulation of ICAM-1 and / or CD18 and / or by suppression of L-selectin surface expression); increased eosinophil viability (e.g., by production of IL-6, eosinophil-derived neurotoxin, and chemokines (C-X-C motif chemokine ligand 8 (CXCL8), CXCL1, and chemokine (C-C motif) ligand 2 (CCL2)); basophil differentiation; production of T2 cytokines by mast cells; production of chemokines CXCL8 and / or CCL1 by mast cells; and macrophage differentiation (e.g., by enhanced expression of the CD80 activation marker).

[0491] In some embodiments, the polypeptide reduces a T2 inflammatory response mediated by one or more innate immune mechanisms of action of TSLP by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the polypeptide reduces a T2 inflammatory response mediated by one or more innate immune mechanisms of action of TSLP by at least about 30%.

[0492] In some embodiments, the polypeptide reduces a T2 inflammatory response mediated by one or more adaptive immune mechanisms of action of TSLP.

[0493] Non-limiting examples of the T2 inflammatory response include increased expression of major histocompatibility complex class II in dendritic cells; increased expression of one or more costimulatory molecules (e.g., CD40, CD86, CD54, CD90, CD83, and / or CD-LAMP) in dendritic cells; increased expression of one or more chemokines (e.g., CXCL8, CCL24, CCL17, CCL22, and CCL1) in dendritic cells; increased expression of OX40 ligand (OX40L); naive T cell differentiation and / or polarization (e.g., naive CD4 +TNF-α from T cells + IL-10 - differentiation into T helper (Th) 2 cells; naive CD4 + T cell proliferation and / or differentiation into Th2 or memory T cells; and CD8 upon TCR stimulation + Increased T cell expansion.

[0494] In some embodiments, the polypeptide reduces a T2 inflammatory response mediated by TSLP's adaptive immune mechanism by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the polypeptide reduces a T2 inflammatory response mediated by TSLP's adaptive immune mechanism by at least about 30%.

[0495] In some embodiments, the polypeptide has less polyspecificity for a non-specific target (e.g., DNA) than a reference antibody. The DNA polyspecificity reagent (PSR) score is a measure of antibody binding to a non-specific DNA target. The DNA PSR score is calculated by dividing the ELISA or DELFIA score by the background score of the secondary antibody alone. A lower DNA PSR score indicates lower polyspecific binding activity and is preferred for the polypeptides of the present invention.

[0496] In some embodiments, the polypeptide has a DNA PSR score of no more than about 1, 2, 3, or 4. In some embodiments, the polypeptide has a DNA PSR score of about 1, 2, 3, or 4. In some embodiments, the polypeptide has a DNA PSR score of about 1-4, 1-3, 1-2, 2-4, 2-3, or 3-4. In some embodiments, the polypeptide has a DNA PSR score of about 2-4, 2-3, or 3-4.

[0497] In some embodiments, the polypeptide has a DNA PSR score that is at least about 10% lower than the reference antibody, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% lower. In some embodiments, the polypeptide has a DNA PSR score that is at least about 30% lower than the reference antibody.

[0498] In some embodiments, the polypeptide has a DNA PSR score that is less than about 90% of the reference antibody, e.g., less than about 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the reference antibody.

[0499] In some embodiments, the polypeptide has a DNA PSR score of about 1 to 90% of that of a reference antibody, for example, about 2 to 90%, 2 to 85%, 3 to 85%, 3 to 80%, 4 to 80%, 4 to 75%, 5 to 75%, 5 to 70%, 6 to 70%, 6 to 65%, 7 to 65%, 7 to 60%, 8 to 60%, 8 to 55%, 9 to 55%, 9 to 50%, 10 to 50%, 10 to 45%, 15 to 45%, 15 to 40%, 20 to 40%, 20 to 35%, 25 to 35%, or 25 to 30% of that of the reference antibody.

[0500] In some embodiments, the polypeptide exhibits less self-association than a reference antibody, as determined, for example, by affinity capture self-interaction nanoparticle spectroscopy (AC-SINS) values. The AC-SINS value is the shift in the maximum absorbance wavelength in the absorption spectrum of the coated nanoparticle compared to the spectrum of the nanoparticle alone. Thus, the greater the shift in the maximum absorbance wavelength, the stronger the self-interaction of the antibody coated on the nanoparticle. Self-association is an undesirable property that correlates with poor viscosity and poor PK properties. Techniques and assays for assessing protein self-association are known in the art. See, e.g., Patro & Przybycien, Biotechnol Bioeng. 52(2):193-203 (1996), the contents of which are incorporated herein by reference in their entirety.

[0501] In some embodiments, the polypeptide has an AC-SINS value not greater than about 0, 1, 2, 3, 4, 5, 6, or 7. In some embodiments, the polypeptide has an AC-SINS value not greater than about 0, 1, 2, 3, or 4. In some embodiments, the polypeptide has an AC-SINS value of about 0, 1, 2, 3, 4, 5, 6, or 7. In some embodiments, the polypeptide has an AC-SINS value of about 0, 1, 2, 3, or 4. In some embodiments, the polypeptide has an AC-SINS value of at least about 0, 1, 2, 3, 4, 5, 6, or 7. In some embodiments, the polypeptide has an AC-SINS value of about 0-4.

[0502] In some embodiments, the polypeptide has an AC-SINS value that is at least about 10% lower than the reference antibody, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% lower than the reference antibody. In some embodiments, the polypeptide has an AC-SINS value that is at least about 30% lower than the reference antibody.

[0503] In some embodiments, the polypeptide has an AC-SINS value that is less than about 90% of that of the reference antibody, e.g., less than about 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of that of the reference antibody.

[0504] In some embodiments, the polypeptide has an AC-SINS value of about 1 to 90% of that of a reference antibody, for example, about 2 to 90%, 2 to 85%, 3 to 85%, 3 to 80%, 4 to 80%, 4 to 75%, 5 to 75%, 5 to 70%, 6 to 70%, 6 to 65%, 7 to 65%, 7 to 60%, 8 to 60%, 8 to 55%, 9 to 55%, 9 to 50%, 10 to 50%, 10 to 45%, 15 to 45%, 15 to 40%, 20 to 40%, 20 to 35%, 25 to 35%, or 25 to 30% of that of the reference antibody.

[0505] fusion proteins The present disclosure also provides, inter alia, fusion proteins comprising one or more of the polypeptides disclosed herein.

[0506] The term "fusion protein" refers to a synthetic, semi-synthetic, or recombinant single protein molecule. A fusion protein can contain all or part of two or more different proteins and / or polypeptides that are linked by a covalent bond (e.g., a peptide bond).

[0507] Fusion proteins of the present disclosure can be produced recombinantly or synthetically using conventional methods and reagents known in the art. For example, fusion proteins of the present disclosure can be produced recombinantly in suitable host cells (e.g., bacteria) according to methods known in the art. See, for example, Current Protocols in Molecular Biology, Second Edition, Ausubel et al. eds., John Wiley & Sons, 1992; and Molecular Cloning: a Laboratory Manual, 2nd edition, Sambrook et al., 1989, Cold Spring Harbor Laboratory Press. For example, a nucleic acid molecule comprising a nucleotide sequence encoding a fusion protein described herein can be introduced into a suitable host cell (e.g., E. coli) and expressed, and the expressed fusion protein can be isolated / purified from the host cell (e.g., in inclusion bodies) using conventional methods and readily available reagents. For example, DNA fragments encoding different protein sequences (e.g., photoreactive domains, heterologous peptide components) can be ligated together in frame according to conventional techniques. In another embodiment, the fusion gene can be synthesized by conventional techniques, including automated DNA synthesizers. Alternatively, anchor primers can be used to PCR amplify nucleic acid fragments, resulting in complementary overhangs between two consecutive nucleic acid fragments, which can then be annealed and replated to generate a chimeric nucleic acid sequence (see Ausubel et al., Current Protocols in Molecular Biology, 1992).

[0508] Nucleic acids, vectors, and host cells The present disclosure also provides, inter alia, one or more polynucleotides (e.g., DNA, RNA, or analogs of either, e.g., optionally containing one or more modified nucleotides; polynucleotides may be linear or circular, e.g., linear or circular RNA) encoding any one of the polypeptides or fusion proteins described herein. In some embodiments, a polypeptide or fusion protein disclosed herein is encoded by a single polynucleotide. In some embodiments, a polypeptide or fusion protein disclosed herein is encoded by multiple polynucleotides.

[0509] In some embodiments, the polynucleotide comprises a nucleotide sequence that is codon-optimized for a host cell of choice.

[0510] The present disclosure also provides, inter alia, vectors (eg, expression vectors, including viral delivery vectors) that include any one or more of the polynucleotides disclosed herein.

[0511] The term "expression vector" refers to a replicable nucleic acid from which one or more proteins can be expressed when the expression vector is transformed into a suitable expression host cell.

[0512] In some embodiments, the vector (e.g., expression vector) further comprises an expression-regulating polynucleotide sequence operably linked to the polynucleotide, a polynucleotide sequence encoding a selectable marker, or both. In some embodiments, the expression-regulating polynucleotide sequence comprises a promoter sequence, an enhancer sequence, or both. In some embodiments, the expression-regulating polynucleotide sequence comprises an inducible promoter sequence. The term "promoter" refers to a region of DNA to which RNA polymerase binds and initiates transcription of a gene. The term "operably linked" means that a nucleic acid is positioned in a recombinant polynucleotide, e.g., vector, such that expression of the nucleic acid is under the control of the element (e.g., promoter) to which it is linked is possible. The term "selectable marker element" is an element that confers a trait suitable for artificial selection. The selectable marker element may be a negative or positive selection marker.

[0513] The present disclosure also provides, inter alia, expression host cells comprising any one or more of the polynucleotides or expression vectors disclosed herein.

[0514] The term "expression host cell" refers to a cell useful for accepting, maintaining, replicating, and / or propagating a vector. Non-limiting examples of expression host cells include mammalian cells such as hybridoma cells, Chinese hamster ovary (CHO) cells, COS cells, human embryonic kidney (HEK), yeast cells such as Pichia pastoris cells, or bacterial cells such as E. coli, including DH5α.

[0515] composition The present disclosure also provides, inter alia, compositions comprising any one of the polypeptides or fusion proteins disclosed herein. In some embodiments, the composition is a pharmaceutical composition.

[0516] In some embodiments, the composition (e.g., pharmaceutical composition) further comprises a pharmaceutically acceptable carrier, excipient, stabilizer, diluent, or isotonic agent (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Suitable pharmaceutically acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the dosages and concentrations employed. Non-limiting examples of pharmaceutically acceptable carriers, excipients, stabilizers, diluents or isotonic agents include buffers (e.g., phosphate, citrate, histidine), antioxidants (e.g., ascorbic acid or methionine), preservatives, proteins (e.g., serum albumin, gelatin or immunoglobulins); hydrophilic polymers, amino acids, carbohydrates (e.g., monosaccharides, disaccharides, glucose, mannose or dextrins); chelating agents (e.g., EDTA), sugars (e.g., sucrose, mannitol, trehalose or sorbitol), salt-forming counterions (e.g., sodium), metal complexes (e.g., Zn-protein complexes); non-ionic surfactants (e.g., Tween®), PLURONICS™ and polyethylene glycol (PEG).

[0517] In some embodiments, the composition (e.g., pharmaceutical composition) is formulated for a suitable administration schedule and route. Non-limiting examples of administration routes include oral, rectal, mucosal, intravenous, intramuscular, subcutaneous, and topical. In some embodiments, the composition (e.g., pharmaceutical composition) is stored in the form of an aqueous solution or a dried formulation (e.g., lyophilized).

[0518] In some embodiments, the composition is formulated to be administered by injection (e.g., intravenous injection). In other embodiments, the composition is formulated for subcutaneous administration.

[0519] In some embodiments, the composition is provided in a dosage form, for example, a pre-filled syringe or an autoinjector.

[0520] In some embodiments, the pharmaceutical composition comprises about 50 mg to about 300 mg of a polypeptide or fusion protein disclosed herein, e.g., about 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 120 mg, 140 mg, 150 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 250 mg, 260 mg, 280 mg, or 300 mg. In some embodiments, the pharmaceutical composition comprises about 60 mg to about 300 mg of a polypeptide or fusion protein disclosed herein, e.g., about 60 to 280 mg, 80 to 280 mg, 80 to 260 mg, 100 to 260 mg, 100 to 250 mg, 120 to 250 mg, 120 to 240 mg, 140 to 240 mg, 140 to 220 mg, 150 to 250 mg, 150 to 200 mg, 160 to 220 mg, 160 to 200 mg, or 180 to 200 mg.

[0521] In some embodiments, the pharmaceutical composition contains about 50 mg / ml to about 200 mg / ml of a polypeptide or fusion protein disclosed herein, e.g., about 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 120 mg / ml, 140 mg / ml, 150 mg / ml, 160 mg / ml, 180 mg / ml, or 200 mg / ml of a polypeptide or fusion protein disclosed herein. In some embodiments, the pharmaceutical composition contains about 60 mg / ml to about 200 mg / ml of a polypeptide or fusion protein disclosed herein, e.g., about 60-180 mg / ml, 80-180 mg / ml, 80-160 mg / ml, 100-160 mg / ml, 100-150 mg / ml, 120-150 mg / ml, or 120-140 mg / ml.

[0522] In some embodiments, the composition is formulated to be administered with a second therapeutic agent as a combination therapy. In some embodiments, the second therapeutic agent comprises a corticosteroid, a beta-agonist (e.g., a long-acting beta-agonist), a muscarinic antagonist, an anti-inflammatory agent, an IL-4 and / or IL-13 antagonist (e.g., an antibody or antigen-binding fragment thereof targeting IL-13, IL-4, or IL-4R), or any combination thereof. In some embodiments, the second therapeutic agent comprises a corticosteroid (e.g., an inhaled corticosteroid). Non-limiting examples of inhaled corticosteroids include beclomethasone dipropionate, budesonide, ciclesonide, fluticasone furoate, fluticasone propionate, mometasone, and mometasone furoate. In some embodiments, the combination therapy comprises a beta-agonist (e.g., a long-acting beta-agonist). Non-limiting examples of long-acting beta-agonists include albuterol sulfate, formoterol fumarate, salmeterol xinafoate, alformoterol tartrate, olodaterol, and combinations thereof. In some embodiments, the combination therapy includes a muscarinic antagonist. In some embodiments, the combination therapy includes an IL-4 and / or IL-13 antagonist (e.g., for the treatment of allergic and / or asthmatic conditions). Non-limiting examples of IL-4 and / or IL-13 antagonists include pitrakinra, an anti-IL-13 antibody or antigen-binding fragment thereof, an anti-IL-4 antibody or antigen-binding fragment thereof, and an anti-IL-4R antibody or antigen-binding fragment thereof. Non-limiting examples of antibodies or antigen-binding fragments thereof include anti-IL-13 antibodies or antigen-binding fragments thereof (e.g., lebrikizumab, tralokinumab, and anrukinzumab), anti-IL-4 antibodies or antigen-binding fragments thereof (e.g., pascolizumab (SB 240683)), and anti-IL-4R antibodies or antigen-binding fragments thereof (e.g., dupilumab).

[0523] In some embodiments, the composition is formulated to be administered in conjunction with an inhaled corticosteroid (ICS), a long-acting beta-agonist (LABA), a long-acting muscarinic antagonist (LAMA), or any combination thereof.

[0524] In some embodiments, the composition (e.g., pharmaceutical composition) comprises: a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 5 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 21 L (AB-2) The polypeptide comprises:

[0525] In some embodiments, the composition (e.g., pharmaceutical composition) comprises: a) V comprising the amino acid sequence of SEQ ID NO: 5 H , and b) V comprising the amino acid sequence of SEQ ID NO: 21 L (AB-2) The polypeptide comprises:

[0526] In some embodiments, the composition (e.g., pharmaceutical composition) comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106; and b) a light chain (AB-2b) comprising the amino acid sequence of SEQ ID NO: 120 The polypeptide comprises:

[0527] In some embodiments, the composition (e.g., pharmaceutical composition) comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92; and b) a light chain (AB-2a) comprising the amino acid sequence of SEQ ID NO: 120 The polypeptide comprises:

[0528] How to use The present disclosure also provides, inter alia, a method for neutralizing TSLP protein (e.g., full-length TSLP protein) in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as an active ingredient, any one of the polypeptides or fusion proteins disclosed herein.

[0529] In some embodiments, neutralizing a TSLP protein in a subject comprises administering to the subject a pharmaceutically acceptable carrier and a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 5 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 21 L (AB-2) and a polypeptide comprising the compound comprising:

[0530] In some embodiments, neutralizing a TSLP protein in a subject comprises administering to the subject a pharmaceutically acceptable carrier and a) V comprising the amino acid sequence of SEQ ID NO: 5 H , and b) V comprising the amino acid sequence of SEQ ID NO: 21 L (AB-2) and a polypeptide comprising the compound comprising:

[0531] In some embodiments, neutralizing a TSLP protein in a subject comprises administering to the subject a pharmaceutically acceptable carrier and a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106; and b) a light chain (AB-2b) comprising the amino acid sequence of SEQ ID NO: 120 and a polypeptide comprising the compound comprising:

[0532] In some embodiments, neutralizing a TSLP protein in a subject comprises administering to the subject a pharmaceutically acceptable carrier and a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92; and b) a light chain (AB-2a) comprising the amino acid sequence of SEQ ID NO: 120 and a polypeptide comprising the compound comprising:

[0533] In some embodiments, the subject is a mammal. In some embodiments, the subject is a mammal selected from the group consisting of a dog, a cat, a mouse, a rat, a hamster, a guinea pig, a horse, a pig, a sheep, a cow, a chimpanzee, a macaque, a cynomolgus monkey, and a human. In some embodiments, the subject is a primate. In some embodiments, the subject is a human.

[0534] In some embodiments, the subject is 18 years of age or older, e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80 years of age or older. In some embodiments, the subject is 65 years of age or older. In some embodiments, the subject is about 18-80, 20-80, 20-75, 25-75, 25-70, 30-70, 30-65, 35-65, 35-60, 40-60, 40-55, or 45-55 years of age. In some embodiments, the subject is about 18-65 years of age.

[0535] In some embodiments, the subject is about 18 years of age or younger. In some embodiments, the subject is about 16, 14, 12, 10, 8, 6, 4, or 2 years of age or older. In some embodiments, the subject is 2 years of age or older, e.g., about 2-18, 3-18, 3-16, 4-16, 4-14, 5-14, 5-12, 6-12, 6-10, or 8-10 years of age.

[0536] In some embodiments, the subject is a pediatric patient. In some embodiments, the subject is a pediatric patient aged 12 years or older.

[0537] In some embodiments, the subject has a blood eosinophil count of less than 150 cells / μL. In some embodiments, the subject has a blood eosinophil count of 150 to less than 300 cells / μL. In some embodiments, the subject has a blood eosinophil count of 300 cells / μL or greater.

[0538] In some embodiments, the subject has or is suspected of having a TSLP-related disease or condition (e.g., a disease or condition associated with dysregulation of TSLP expression, such as overexpression of TSLP). In some embodiments, the subject has a TSLP-related disease or condition. In some embodiments, the subject has been diagnosed with a TSLP-related disease or condition. In other embodiments, the subject is at risk of developing a TSLP-related disease or condition.

[0539] Non-limiting examples of TSLP-associated diseases and / or conditions (e.g., diseases and / or conditions associated with dysregulated TSLP expression) include asthma, AD, allergic conjunctivitis, chronic obstructive pulmonary disease (COPD), chronic spontaneous urticaria (CSU), nasal polyps, rheumatoid arthritis (RA), rhinosinusitis (RS) (e.g., allergic rhinosinusitis, severe chronic rhinosinusitis (e.g., severe chronic rhinosinusitis with nasal polyps)), cancer, and food hypersensitivity reactions. Non-limiting examples of TSLP-associated cancers (e.g., cancers associated with dysregulated TSLP expression) include lymphoma, acute lymphocytic leukemia (ALL), multiple sclerosis, and solid tumors (e.g., cervical cancer, ovarian cancer, pancreatic cancer, gastric cancer, or colorectal cancer (e.g., colon cancer, rectal cancer, and variants thereof)).

[0540] In some embodiments, the subject has an inflammatory disorder (e.g., an allergic inflammatory disorder). In some embodiments, the subject has a pulmonary inflammatory disease or condition. In some embodiments, the subject has asthma (e.g., severe and / or uncontrolled asthma). In some embodiments, the subject has mild, moderate, moderate-to-severe, or severe asthma. In some embodiments, the subject has uncontrolled asthma (e.g., uncontrolled moderate asthma, uncontrolled moderate-to-severe asthma, or uncontrolled severe asthma), allergic asthma (e.g., mild allergic asthma, moderate allergic asthma, or severe allergic asthma), oral corticosteroid-dependent asthma, or any combination thereof.

[0541] In some embodiments, the subject has been previously treated with one or more therapeutic agents before being administered the composition (e.g., a polypeptide or pharmaceutical composition disclosed herein).

[0542] In some embodiments, the subject has previously received a therapeutic or prophylactic medication, for example, a COPD patient may have previously received an inhaled corticosteroid (ICS), a long-acting beta-agonist (LABA), a long-acting muscarinic antagonist (LAMA), or any combination thereof.

[0543] The present disclosure also provides, inter alia, a method of treating a TSLP-associated disease or condition in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as an active ingredient, any one of the polypeptides or fusion proteins disclosed herein.

[0544] In some embodiments, treating a TSLP-associated disease or condition in a subject comprises administering to the subject a TSLP-associated disease or condition comprising administering to the subject a pharmaceutically acceptable carrier and a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 5 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 21 L (AB-2) and a polypeptide comprising the compound comprising:

[0545] In some embodiments, treating a TSLP-associated disease or condition in a subject comprises administering to the subject a TSLP-associated disease or condition comprising administering to the subject a pharmaceutically acceptable carrier and a) V comprising the amino acid sequence of SEQ ID NO: 5 H , and b) V comprising the amino acid sequence of SEQ ID NO: 21 L (AB-2) and a polypeptide comprising the compound comprising:

[0546] In some embodiments, treating a TSLP-associated disease or condition in a subject comprises administering to the subject a TSLP-associated disease or condition comprising administering to the subject a pharmaceutically acceptable carrier and a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106; and b) a light chain (AB-2b) comprising the amino acid sequence of SEQ ID NO: 120 and a polypeptide comprising the compound comprising:

[0547] In some embodiments, treating a TSLP-associated disease or condition in a subject comprises administering to the subject a TSLP-associated disease or condition comprising administering to the subject a pharmaceutically acceptable carrier and a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92; and b) a light chain (AB-2a) comprising the amino acid sequence of SEQ ID NO: 120 and a polypeptide comprising the compound comprising:

[0548] In some embodiments, treating a TSLP-related disease or condition includes treating asthma (e.g., severe and / or uncontrolled asthma). In some embodiments, treating a TSLP-related disease or condition includes treating moderate to severe asthma. In some embodiments, treating a TSLP-related disease or condition includes treating severe asthma. In some embodiments, treating a TSLP-related disease or condition includes treating uncontrolled asthma. In some embodiments, treating a TSLP-related disease or condition includes treating severe uncontrolled asthma. In some embodiments, treating a TSLP-related disease or condition includes treating AD.

[0549] In some embodiments, treating a TSLP-associated disease or condition is a prophylactic therapy.

[0550] In some embodiments, the effective amount is sufficient to prevent the subject from developing a TSLP-associated disease or condition (eg, a disease or condition associated with dysregulated TSLP expression).

[0551] In some embodiments, an effective amount is sufficient to reduce TSLPR occupancy in a subject, ie, by at least about 10%, e.g., by at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, an effective amount is sufficient to reduce TSLPR occupancy by about 10-99%, e.g., about 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0552] In some embodiments, an effective amount is sufficient to reduce the TSLP receptor complex in a subject, ie, by at least about 10%, e.g., by at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, an effective amount is sufficient to reduce the TSLP receptor complex by about 10-99%, e.g., about 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0553] In some embodiments, an effective amount is sufficient to reduce the activity of a TSLP protein in a subject. In some embodiments, an effective amount is sufficient to reduce the activity of a TSLP protein by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, an effective amount is sufficient to reduce the activity of a TSLP protein by about 10-99%, e.g., about 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0554] In some embodiments, an effective amount is sufficient to reduce airway inflammation (e.g., the number and / or density of one or more types of airway submucosal inflammatory cells) in a subject. In some embodiments, an effective amount is sufficient to reduce airway inflammation by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, an effective amount is sufficient to reduce airway inflammation by about 10-99%, e.g., about 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0555] In some embodiments, an effective amount is sufficient to reduce airway remodeling (e.g., reticular basement membrane thickening, airway epithelial integrity) in a subject. In some embodiments, an effective amount is sufficient to reduce airway remodeling by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, an effective amount is sufficient to reduce airway remodeling by about 10-99%, e.g., about 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0556] In some embodiments, the effective amount is sufficient to treat nasal polyps in a subject (e.g., reduce the size and / or number of nasal polyps). In some embodiments, the effective amount is sufficient to reduce the size of nasal polyps by at least about 10%, e.g., by at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the effective amount is sufficient to reduce the size of nasal polyps by about 10-99%, e.g., about 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%. In some embodiments, the effective amount is sufficient to reduce the number of nasal polyps by at least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the effective amount is sufficient to reduce the number of nasal polyps by about 10-99%, e.g., about 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0557] The therapeutic agents disclosed herein can be administered by a variety of routes of administration, depending on the compound and the particular condition to be treated, including, for example, topical, transdermal, parenteral (e.g., intra-arterial, intravenous, intramuscular, subcutaneous injection, intradermal injection), intravenous infusion, and inhalation (e.g., intrabronchial, intranasal or oral inhalation, nasal drops) routes of administration.

[0558] Administration can be local or systemic depending on the indication. The preferred mode of administration can vary depending on the particular compound selected.

[0559] In some embodiments, a composition (eg, a polypeptide or pharmaceutical composition) is administered to a subject as a monotherapy.

[0560] In some embodiments, a composition (e.g., a polypeptide or pharmaceutical composition) is administered to a subject in combination with one or more additional therapeutic agents (e.g., simultaneously with or subsequent to one or more additional therapeutic agents) or in combination with a prophylactic agent (e.g., simultaneously with or subsequent to one or more prophylactic agents).

[0561] In some embodiments, treating a TSLP-associated disease or condition further comprises administering to the subject a therapeutically effective amount of one or more additional therapeutic agents at the same time as, or after, administration of the composition (e.g., polypeptide or pharmaceutical composition). In some embodiments, treating a TSLP-associated disease or condition further comprises administering to the subject a therapeutically effective amount of one or more prophylactic agents before, at the same time as, or after administration of the composition (e.g., polypeptide or pharmaceutical composition).

[0562] Administration of two or more therapeutic agents includes co-administration of the therapeutic agents in a substantially simultaneous manner, such as a pharmaceutical combination. Alternatively, such administration includes co-administration of each therapeutic agent in multiple or separate containers (e.g., capsules, powders, and liquids). Such administration also includes use of each type of therapeutic agent in a sequential manner, either at about the same time or at different times. The composition and the second therapeutic agent can be administered by the same route of administration or by different routes of administration.

[0563] The present disclosure also provides, inter alia, methods for reducing (e.g., inhibiting) the binding of TSLP to TSLPR in a cell, comprising contacting the cell with an effective amount of any one or more of the polypeptides or fusion proteins disclosed herein.

[0564] In some embodiments, reducing (e.g., inhibiting) binding of TSLP to the cellular TSLPR includes: a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 5 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 21 L (AB-2) The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0565] In some embodiments, reducing (e.g., inhibiting) binding of TSLP to the cellular TSLPR includes: a) V comprising the amino acid sequence of SEQ ID NO: 5 H , and b) V comprising the amino acid sequence of SEQ ID NO: 21 L (AB-2) The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0566] In some embodiments, reducing (e.g., inhibiting) binding of TSLP to the cellular TSLPR includes: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106; and b) a light chain (AB-2b) comprising the amino acid sequence of SEQ ID NO: 120 The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0567] In some embodiments, reducing (e.g., inhibiting) binding of TSLP to the cellular TSLPR includes: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92; and b) a light chain (AB-2a) comprising the amino acid sequence of SEQ ID NO: 120 The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0568] The present disclosure also provides, inter alia, a method for inhibiting TSLP-induced signaling in a cell, the method comprising contacting the cell with an effective amount of any one of the polypeptides or fusion proteins disclosed herein.

[0569] In some embodiments, inhibiting TSLP-induced signaling in a cell includes: a) a V comprising the HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NO: 5 H and b) a V comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NO: 21 L (AB-2) The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0570] In some embodiments, inhibiting TSLP-induced signaling in a cell includes: a) V comprising the amino acid sequence of SEQ ID NO: 5 H , and b) V comprising the amino acid sequence of SEQ ID NO: 21 L (AB-2) The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0571] In some embodiments, inhibiting TSLP-induced signaling in a cell includes: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 106; and b) a light chain (AB-2b) comprising the amino acid sequence of SEQ ID NO: 120 The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0572] In some embodiments, inhibiting TSLP-induced signaling in a cell includes: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 92; and b) a light chain (AB-2a) comprising the amino acid sequence of SEQ ID NO: 120 The method comprises contacting the cell with an effective amount of a polypeptide comprising:

[0573] In some embodiments, the cells are selected from the group consisting of airway smooth muscle cells (ASMC), basophils, dendritic cells, eosinophils, type 2 innate lymphoid cells (ILC2), hematopoietic progenitor cells, lymphocytes, macrophages, mast cells, and monocytes.

[0574] AB-2 The present disclosure also provides, inter alia: a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 31, HCDR2 comprising the amino acid sequence of SEQ ID NO: 35, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 50; and b) LCDR1 comprising the amino acid sequence of SEQ ID NO: 60, LCDR2 comprising the amino acid sequence of SEQ ID NO: 67, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0575] The present disclosure also provides, inter alia: a) HCDR1 consisting of the amino acid sequence of SEQ ID NO: 31, HCDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and HCDR3 consisting of the amino acid sequence of SEQ ID NO: 50; and b) LCDR1 consisting of the amino acid sequence of SEQ ID NO: 60, LCDR2 consisting of the amino acid sequence of SEQ ID NO: 67, and LCDR3 consisting of the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0576] The AB-2 CDR amino acid sequence, as determined by Kabat numbering, is: HCDR1: TYGMH (SEQ ID NO: 130) HCDR2: VVWYDGSYTHYADSVKG (SEQ ID NO: 131) HCDR3: SPQWEEIFEAMDI (SEQ ID NO: 132) LCDR1:GGNNIGSKSVH (SEQ ID NO: 133) LCDR2: DDSDRPS (SEQ ID NO: 134) LCDR3: QIWDSSSSLVV (SEQ ID NO: 71) is.

[0577] The present disclosure also provides, inter alia: a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 130, HCDR2 comprising the amino acid sequence of SEQ ID NO: 131, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 132; and b) LCDR1 comprising the amino acid sequence of SEQ ID NO: 133, LCDR2 comprising the amino acid sequence of SEQ ID NO: 134, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0578] The present disclosure also provides, inter alia: a) HCDR1 consisting of the amino acid sequence of SEQ ID NO: 130, HCDR2 consisting of the amino acid sequence of SEQ ID NO: 131, and HCDR3 consisting of the amino acid sequence of SEQ ID NO: 132; and b) LCDR1 consisting of the amino acid sequence of SEQ ID NO: 133, LCDR2 consisting of the amino acid sequence of SEQ ID NO: 134, and LCDR3 consisting of the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0579] The AB-2 CDR amino acid sequence, as determined by Chothia numbering, is: HCDR1: GFTFRTY (SEQ ID NO: 135) HCDR2: WYDGSY (SEQ ID NO: 136) HCDR3: SPQWEEIFEAMDI (SEQ ID NO: 132) LCDR1:GGNNIGSKSVH (SEQ ID NO: 133) LCDR2: DDSDRPS (SEQ ID NO: 134) LCDR3: QIWDSSSSLVV (SEQ ID NO: 71) is.

[0580] The present disclosure also provides, inter alia: a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 135, HCDR2 comprising the amino acid sequence of SEQ ID NO: 136, and HCDR3 comprising the amino acid sequence of SEQ ID NO: 132; and b) LCDR1 comprising the amino acid sequence of SEQ ID NO: 133, LCDR2 comprising the amino acid sequence of SEQ ID NO: 134, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0581] The present disclosure also provides, inter alia: a) HCDR1 consisting of the amino acid sequence of SEQ ID NO: 135, HCDR2 consisting of the amino acid sequence of SEQ ID NO: 136, and HCDR3 consisting of the amino acid sequence of SEQ ID NO: 132; and b) LCDR1 consisting of the amino acid sequence of SEQ ID NO: 133, LCDR2 consisting of the amino acid sequence of SEQ ID NO: 134, and LCDR3 consisting of the amino acid sequence of SEQ ID NO: 71 A polypeptide comprising: Also provided are polypeptides that are antibodies or antigen-binding fragments thereof.

[0582] In some embodiments, the polypeptide is a) V that is humanized, contains human framework regions, or a combination thereof H ; b) V that is humanized, contains human framework regions, or a combination thereof L , Or both a) and b) Includes.

[0583] In some embodiments, the polypeptide is a) V comprising the amino acid sequence of SEQ ID NO: 5 H ; b) V comprising the amino acid sequence of SEQ ID NO: 21 L , Or both a) and b) Includes.

[0584] In some embodiments, the antigen-binding fragment is a single-chain fragment variable (scFv), a variable heavy chain domain of the heavy chain (V HH ), fragment antigen binding (Fab), Fab' or F(ab')2.

[0585] In some embodiments, the polypeptide is a) an antibody heavy chain constant domain; b) an antibody light chain constant domain; Or both a) and b) Includes.

[0586] In some embodiments, the antibody heavy chain constant domain is an IgG1, IgG2, IgG3, or IgG4 constant domain. In some embodiments, the antibody heavy chain constant domain is an IgG1 or IgG2 constant domain. In some embodiments, the antibody heavy chain constant domain comprises one or more mutations that increase the serum half-life of the antibody or antigen-binding fragment thereof in humans. In some embodiments, the antibody heavy chain constant domain comprises amino acid substitutions with tyrosine, threonine, and glutamic acid at amino acid residues 252, 254, and 256, respectively (wherein amino acid residues are numbered according to the EU index as in Kabat), compared to a wild-type human IgG constant domain.

[0587] In some embodiments, the polypeptide is a) an antibody heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 92; b) an antibody light chain (LC) comprising the amino acid sequence of SEQ ID NO: 120; Or both a) and b) Includes.

[0588] In some embodiments, the polypeptide is a) an antibody heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 106; b) an antibody light chain (LC) comprising the amino acid sequence of SEQ ID NO: 120; Or both a) and b) Includes.

[0589] In some embodiments, the polypeptide specifically binds to TSLP.

[0590] The present disclosure also provides compositions comprising, inter alia, one or more polypeptides disclosed in the section entitled "AB-2" and one or more pharmaceutical excipients, diluents, or carriers.

[0591] In some embodiments, the composition further comprises one or more additional therapeutic agents, hi some embodiments, the one or more additional therapeutic agents comprise a corticosteroid, a beta agonist, a muscarinic antagonist, an anti-inflammatory agent, an IL-4 and / or IL-13 antagonist (e.g., an antibody or antigen-binding fragment thereof that targets IL-13, IL-4, or IL-4R), or a combination thereof.

[0592] The present disclosure also provides, inter alia, a method of treating a subject in need thereof, comprising administering an effective amount of a composition disclosed in the section entitled "AB-2."

[0593] In some embodiments, the subject has asthma, atopic dermatitis (AD), allergic conjunctivitis, chronic obstructive pulmonary disease (COPD), chronic spontaneous urticaria (CSU), rheumatoid arthritis (RA), rhinosinusitis (RS), eosinophilic esophagitis (EE), or food hypersensitivity. In some embodiments, the subject has moderate asthma. In some embodiments, the subject has severe asthma. In some embodiments, the subject has COPD. In some embodiments, the subject has COPD and / or either moderate asthma or severe asthma.

[0594] In some embodiments, the method further includes administering to the subject an effective amount of a corticosteroid, a beta agonist, a muscarinic antagonist, an anti-inflammatory agent, an IL-4 and / or IL-13 antagonist (e.g., an antibody or antigen-binding fragment thereof that targets IL-13, IL-4, or IL-4R), or a combination thereof.

[0595] The present disclosure also provides, inter alia, a method for reducing binding of TSLP to TSLPR on cells of a subject, the method comprising contacting the cells with an effective amount of a composition disclosed in the section entitled "AB-2."

[0596] The present disclosure also provides, inter alia, fusion proteins comprising the polypeptides disclosed in the section entitled "AB-2."

[0597] The present disclosure also provides, inter alia, a polynucleotide comprising a nucleotide sequence that encodes a polypeptide disclosed in the section entitled "AB-2."

[0598] The present disclosure also provides, inter alia, host cells comprising the polynucleotides disclosed in the section entitled "AB-2."

[0599] The present disclosure also provides, inter alia, a method of making a polypeptide disclosed in the section entitled "AB-2," comprising culturing a host cell containing a nucleotide sequence encoding the polypeptide under conditions whereby the polypeptide is expressed in the host cell.

[0600] Computer-aided sequence definition and validation Applicants have facilitated an understanding of the structure-function interrelationships of anti-TSLP antibodies through computational design and experimental validation, and herein provide those skilled in the art with the means to employ such understanding. For example, the present application provides computer program listing appendices (also referred to herein as Appendix A and Appendix B, respectively) that can be used to both score (score) a given sequence or generate sequences that, upon scoring, have a score above a given threshold. These appendices include Potts models (generalized Ising models) of the polypeptides provided by the present invention. In some embodiments, implementations of the Potts model assume that amino acids interact with their nearest neighbors, but relationships with distant amino acids (e.g., those located two or more positions apart, thus representing features of secondary and tertiary structure) can be determined as well.

[0601] A Potts model is a quadratic sequence model. In some embodiments, the Potts model is energy-based. The model is designed to output the energy for a particular configuration. Given a sequence provided to the energy-based model, the model can be trained to determine the quantification of any measure. The Potts model is represented by a model weight table. This table includes primary and secondary weights. For primary weights, the table includes a residue position, a residue (e.g., amino acid), and a score associated with the residue at that position. For quadratic weights, the table includes two positions, two residues, and one score for the two residues at each position. The score is the self-energy or energy score.

[0602] The computer program listing appendices are referred to as Appendix A (claim.py) and Appendix B (fit_model.etab), which are available online at the respective addresses listed above and are incorporated herein by reference in their entirety. Those skilled in the art will recognize that Appendix A can be executed in a Python environment, compiler, or other equivalent environment capable of executing Python scripts. Those skilled in the art will recognize that Appendix B can be loaded by the script in Appendix A. In an embodiment, the table entries in Table 15 in Appendix D may be copied into an American Standard Code for Information Interchange (ASCII) document. In an embodiment, those skilled in the art will recognize that the script "claim.py" in Appendix C is substantially similar to that of the file "claim.py" in Appendix A (incorporated by reference in its entirety). The claim.py file can be executed to input two sequences (or one combined sequence). In an embodiment, the following description further describes the source code and models in Appendix A / Appendix C and Appendix B / D.

[0603] FIG. 12A is a diagram 1000 illustrating zero-order, first-order, and second-order models. The zero-order model 1002 is an unordered collection of amino acids. Because this model is zero-order, there is no order or structure to the amino acids. A physical analogy can be thought of as a "bag" of amino acids, where the amino acids are jumbled and in no order. The first-order model 1004 is then a matrix represented as per-residue quantifications of the protein sequence (e.g., peptide mass fingerprints (PMFs) of amino acids). Each row of the matrix is an amino acid sequence, and each column is constructed so that the quantifications of each residue in that column sum to 1. These first-order energies can be referred to as the h-tensor, hereafter referred to as the "first hash table." The first-order terms of the model are said to be at a given position, and the model can assign a score to each specific position. For paired terms, the model can compare the scores given to any two positions. The model can output an "energy," which is a number; in embodiments, the smaller the number, the better.

[0604] The secondary sequence model 1006, or Potts model, represents the energy of two residues viewed together and the energy associated with the primary effects. The secondary energy of two residues viewed together is considered the j-tensor, while the energy associated with the primary effects is referred to as the h-tensor. In other words, the secondary energy compares the energy between two positions in an amino acid sequence. Thus, this secondary alignment compares the energy between two positions in any amino acid sequence. One skilled in the art will recognize that this can be repeated for multiple sequences, thereby scoring many similar sequences, all residue pairs in similar sequences.

[0605] FIG. 12B is a flow diagram 1050 illustrating an example process used by embodiments of the present disclosure to determine claimed sequences. Initially, the process receives a given polypeptide sequence or sequence pair (1052). The sequence pair may be separate polypeptide chains in different embodiments, or may be a single polypeptide chain, e.g., an scFV, in some embodiments. Those skilled in the art will further recognize that, in embodiments, the model in Appendix B (or Appendix D) can be used to generate polypeptide sequences or sequence pairs for testing using the script in Appendix A (or Appendix C). The script in Appendix A (or Appendix C) scores the sequence using the model weights provided in Appendix B (or Appendix D) (1054). The script then determines whether the score exceeds a predetermined threshold (however, those skilled in the art will recognize that in different scoring regimes, the score may be below the threshold) (1056). If not, the polypeptide sequence or sequence pair is not validated (1058). If so, the polypeptide sequence or sequence pair is validated (1060).

[0606] The Potts model disclosed herein was generated from experimental validation of approximately 100 sequences to identify sequences with advantageous functions. The model coefficients are stored in an ASCII text file [fit_model.etab]. When assayed, these sequences exhibited acceptable multispecific reactivity, autoaggregation tendency, and expression. In addition, they all possessed biological function as determined by the SEAP reporter assay. The resulting model was validated to claim only sequences not previously reported in the literature.

[0607] In some embodiments, a script is utilized to identify pairs of sequences claimed under the model (e.g., V H and V LFor clarity, however, it is determined whether a pair of sequences (which may be separate polypeptide chains in different embodiments, or a single polypeptide chain, e.g., scFV, in some embodiments) exceeds a threshold when scored. This script is referred to as a computationally binding optimizing (CBO) script. Two hash tables are loaded from existing files, and the script extracts primary and secondary index / position values, residue values, and the corresponding score for each. For the primary table, the script extracts one position, one residue, and one score associated with the residue at that position. For the secondary table, the script extracts the first position, the second position, the first residue, the second residue, and the scores associated with the first residue at the first position and the second residue at the second position.

[0608] Two sequences, V H Sequence and V L The sequences are input into the script and then aligned. H Sequence and V L It is confirmed that the sequence is the same length as one or more reference sequences, and that both sequences are identical to one or more reference sequences over a given interval. After confirmation, residues are extracted from both input sequences at specified positions to form a trimmed concatenated sequence. The positions used to extract residues are positions where mutations were applied during model training. The positions used to confirm that a given interval of both sequences is identical to one or more reference sequences are positions where no mutations were applied during model training. After trimming to the positions used to train the model, the two input sequences are concatenated together. An energy score is then calculated for the given concatenated sequence and two hash tables. The following steps describe some embodiments for calculating the energy score.

[0609] First,

number

number

number

number

number

number

number

number

number

[0610] The score_sequence function in the script in Appendix A (or Appendix C) calculates and returns the above value. The return value of the score_sequence function represents the log-probability that the sequence will appear given the Potts model weights input to the script. If the score is greater than a threshold (-7.7 in this embodiment), then the sequence can meet the required parameters (e.g., can be confirmed / verified by the Potts model) and the function returns true. This score represents log-probability. Otherwise, the function returns false, and the sequence pair is not confirmed by the model. When the function returns true, it indicates that the sequence pair is suitable for a given purpose (e.g., for binding to human TSLP).

[0611] Those skilled in the art will recognize that other thresholds representing the log probability that a sequence will meet the required parameters can be utilized. In some embodiments, exemplary threshold ranges calculated by the script may include approximately -7.7, -7.0, -6.5, -6.0, -5.5, -5.0, -4.5, -4.0, -3.5, -3.0, 2.9, -2.8, -2.7, -2.6, -2.5, -2.4, -2.3, -2.2, -2.1, -2.0, -1.9, -1.8, -1.7, -1.6, -1.5, -1.4, -1.3, -1.2, -1.1, -1.0, 0.9, -0.8, -0.7, -0.6, -0.5, -0.4, -0.3, -0.2, and -0.1. As the thresholds increase toward zero, sequence spaces that score within those thresholds represent smaller sequence spaces with higher confidence levels. In some embodiments, constants in the script or hash table may be changed, such as by multiplying all constants by a constant. In such cases, thresholds may be adjusted as can be determined by one of skill in the art. Thresholds may also be calculated in a non-logarithmic manner and expressed as confidence intervals, or non-logarithmic ranges, or as other probabilistic thresholds.

[0612] In some embodiments, the script in Appendix A defines a sequence scoring function (sequence_energy). This function calculates the sequence energy of substituting a residue (e.g., an amino acid) for each of the other possible residues at a set of positions in a sequence, using a Potts model trained on the replacing residues at those positions. As described above, scoring checks each possible residue at each required position. Those skilled in the art will recognize that the script can also substitute a residue at each given position (as in the script in Appendix A). In some embodiments, a similar script could add a residue to a sequence. Each score is calculated with the sequence_energy function and added to a score array. For each position examined, the sum of the scores in the array is summed and normalized with a softmax function. The scores are then summed logarithmically and returned.

[0613] In some embodiments, the sequence energy function takes as input a sequence and a hash table determined from an input etab file (or another format). The function iterates through all primary combinations and scores them with the first hash table, and iterates through all secondary combinations and scores them with the second hash table. The sequence_energy function considers all substituted residues and their primary and secondary combinations and sums their energies together.

[0614] In some embodiments, the etab file is read with the read_etab function. Each line in the file with three terms is unpacked and converted into an entry in a first hash table for the primary combination. Each line in the file with five terms is unpacked and converted into an entry in a second hash table for the secondary combination. These two hash tables are returned.

[0615] The steps and functions described above describe "computational bind optimization" (CBO) sequences, amino acid sequences, or polypeptide sequences. Such sequences are sequences that return a true value when input into the computational bind optimization script described above. Those skilled in the art will recognize that the script can be implemented in other ways or with other sequences of steps. Those skilled in the art will also recognize that other functionally equivalent tables and constants can be used. However, the provided Python script, tables, and constants are configured to output whether a given polypeptide sequence is a CBO sequence when executed by a processor. Thus, any sequence that produces a Boolean "true" output when executed by the script is considered a CBO sequence. Any sequence that produces a Boolean "false" output when executed by the script is considered to be excluded from the group of CBO sequences. Thus, in some embodiments, the script in Appendix A (or Appendix C) does not itself determine a CBO sequence, but is a tool for verifying whether a given sequence is a CBO sequence. When a CBO sequence is an amino acid sequence, the corresponding molecule having that amino acid sequence is referred to as a CBO polypeptide.

[0616] In some embodiments, a similar script may return the calculated score for a given sequence or sequences in addition to, or alternatively to, a Boolean value representing a threshold.

[0617] In some embodiments, a similar script can return one or more sequences generated from a Potts model. To generate the sequences, Markov Chain Monte Carlo [MCMC] sampling can be performed on the model. The resulting samples can then be reintegrated into a constant template. In some embodiments, a brute force approach can be applied using a scoring function.

[0618] With the above in mind, Table 10 illustrates the trimmed sequences (vlt, vht), concatenated / combined sequences (vhvlt) of proteins AB-1 to AB-14, and the resulting Potts model scores when the script is applied to them. The sequences of AB-1 to AB-14 are shown in Tables 1 and 2. Proteins AB-1 to AB-14 all had Potts model scores above -7.7 and therefore have been or could be validated by the script.

[0619] In light of Table 10, one skilled in the art will recognize that equivalent Potts model scores, such as scores with different scales or scores multiplied by negative scores, may result in scores at, above, or below different thresholds.

[0620] FIG. 13 illustrates an example computer network or similar digital processing environment in which embodiments of the present invention may be implemented.

[0621] One or more client computers / devices 50 and one or more server computers 60 provide processing, storage, and input / output devices for executing application programs and the like. One or more client computers / devices 50 may also be linked to other computing devices, including other client devices / processes 50 and one or more server computers 60, through a communications network 70. The communications network 70 may be part of a remote access network, a global network (e.g., the Internet), a collection of computers worldwide, a local area or wide area network, and gateways that currently communicate with each other using their respective protocols (TCP / IP, Bluetooth, etc.). Other electronic device / computer network architectures are also suitable.

[0622] FIG. 14 is a diagram of an exemplary internal structure of a computer (e.g., client processor / device 50 or server computer 60) in the computer system of FIG. 13. Each computer 50, 60 includes a system bus 79, where a bus is a set of hardware lines used to transfer data between components of a computer or processing system. The system bus 79 is essentially a shared conduit connecting different elements of a computer system (e.g., processor, disk storage, memory, input / output ports, network ports, etc.), allowing information to be transferred between the elements. Attached to the system bus 79 is an input / output device interface 82 for connecting various input and output devices (e.g., keyboard, mouse, display, printer, speakers, etc.) to the computer 50, 60. A network interface 86 allows the computer to connect to various other devices attached to a network (e.g., network 70 of FIG. 13). Memory 90 provides volatile storage of computer software instructions 92 and data 94 used to implement some embodiments of the present invention (e.g., the CBO decision script, Potts model, and model weight code detailed above and in Appendices A and B). Disk storage 95 provides non-volatile storage of computer software instructions 92 and data 94 used to implement some embodiments of the present invention. Central processing unit 84 is also attached to system bus 79 and provides the execution of computer instructions.

[0623] In one embodiment, the processor routines 92 and data 94 are a computer program product (generally designated by the reference numeral 92) including a non-transitory computer-readable medium (e.g., removable storage medium such as one or more DVD-ROMs, CD-ROMs, diskettes, tapes, etc.) that provides at least a portion of the software instructions for the system of the present invention. The computer program product 92 can be installed by any suitable software installation procedure, as known in the art. In another embodiment, at least a portion of the software instructions may also be downloaded via cable communication and / or wireless connection. In some embodiments, the program of the present invention is a computer program propagated signal product embodied in a propagated signal on a propagated medium (e.g., radio waves, infrared waves, laser waves, sound waves, or electric waves propagating over a global network such as the Internet or one or more other networks). Such a carrier medium or signal may be utilized to provide at least a portion of the software instructions of the routines / program 92 of the present invention.

[0624] In another aspect, the disclosure provides a polypeptide that specifically binds to TSLP (e.g., hTSLP), wherein the CBO model outputs a score above a predetermined threshold when scoring the amino acid sequence of the polypeptide.

[0625] In another aspect, the disclosure provides a CBO polypeptide that specifically binds to TSLP (e.g., hTSLP), wherein the CBO model outputs a score above a predetermined threshold when scoring an amino acid sequence representing the CBO polypeptide.

[0626] In another aspect, the disclosure provides a polypeptide that specifically binds to TSLP (e.g., hTSLP) and receives a score above a predetermined threshold from a CBO model when scoring an amino acid sequence representing the polypeptide.

[0627] In another aspect, the disclosure provides a polypeptide that specifically binds to TSLP (e.g., hTSLP), wherein the polypeptide receives a score above a predetermined threshold from a CBO model when scoring an amino acid sequence representing the polypeptide.

[0628] In another aspect, the disclosure provides computationally optimized (CBO) polypeptides determined by a computational binding optimization (CBO) model that specifically bind to TSLP (e.g., hTSLP).

[0629] In some embodiments, the CBO polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of the above The antibody comprises a paratope substantially similar to the paratope of the antibody comprising an amino acid sequence selected from:

[0630] In some embodiments, the CBO polypeptide is a V selected from SEQ ID NO:4 / SEQ ID NO:20 (AB-1), SEQ ID NO:5 / SEQ ID NO:21 (AB-2), SEQ ID NO:6 / SEQ ID NO:20 (AB-3), SEQ ID NO:7 / SEQ ID NO:22 (AB-4), SEQ ID NO:8 / SEQ ID NO:23 (AB-5), SEQ ID NO:9 / SEQ ID NO:24 (AB-6), SEQ ID NO:10 / SEQ ID NO:25 (AB-7), SEQ ID NO:11 / SEQ ID NO:20 (AB-8), SEQ ID NO:12 / SEQ ID NO:20 (AB-9), SEQ ID NO:13 / SEQ ID NO:26 (AB-10), SEQ ID NO:14 / SEQ ID NO:27 (AB-11), SEQ ID NO:15 / SEQ ID NO:28 (AB-12), SEQ ID NO:16 / SEQ ID NO:29 (AB-13), or SEQ ID NO:17 / SEQ ID NO:30 (AB-14). H / V L The combination of the above includes a paratope that is substantially similar (e.g., has at least about 90% sequence identity; has 100% sequence identity) to the paratope of the combination of the above.

[0631] In some embodiments, the CBO polypeptide is a V selected from SEQ ID NO:4 / SEQ ID NO:20 (AB-1), SEQ ID NO:5 / SEQ ID NO:21 (AB-2), SEQ ID NO:6 / SEQ ID NO:20 (AB-3), SEQ ID NO:7 / SEQ ID NO:22 (AB-4), SEQ ID NO:8 / SEQ ID NO:23 (AB-5), SEQ ID NO:9 / SEQ ID NO:24 (AB-6), SEQ ID NO:10 / SEQ ID NO:25 (AB-7), SEQ ID NO:11 / SEQ ID NO:20 (AB-8), SEQ ID NO:12 / SEQ ID NO:20 (AB-9), SEQ ID NO:13 / SEQ ID NO:26 (AB-10), SEQ ID NO:14 / SEQ ID NO:27 (AB-11), SEQ ID NO:15 / SEQ ID NO:28 (AB-12), SEQ ID NO:16 / SEQ ID NO:29 (AB-13), or SEQ ID NO:17 / SEQ ID NO:30 (AB-14). H / V L and paratopes that differ from the paratopes of the combination by 1 to 3 (e.g., 1, 2, or 3) residue substitutions (e.g., conservative substitutions, such as highly conservative).

[0632] In particular embodiments, the CBO polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1), SEQ ID NO: 5 and SEQ ID NO: 21 (AB-2), SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3), SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), SEQ ID NO: 8 and SEQ ID NO: 23 (AB-5), SEQ ID NO: 9 and SEQ ID NO: 24 (AB-6), SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7), SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8), SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9), SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10), SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11), SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12), SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13), SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), and A combination of these V to be selected from H and V L Does not include pairs.

[0633] In certain embodiments, the CBO polypeptide is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 6 and SEQ ID NO: 20 (AB-3); or SEQ ID NO: 7 and SEQ ID NO: 22 (AB-4), or A combination of the above The antibody comprises a paratope substantially similar to the paratope of the antibody comprising an amino acid sequence selected from:

[0634] In some embodiments, the CBO polypeptide is a V selected from SEQ ID NO:4 / SEQ ID NO:20 (AB-1), SEQ ID NO:5 / SEQ ID NO:21 (AB-2), SEQ ID NO:6 / SEQ ID NO:20 (AB-3), or SEQ ID NO:7 / SEQ ID NO:22 (AB-4). H / V Land a paratope substantially similar (e.g., having at least about 90% sequence identity; 100% sequence identity) to the paratope ...

Claims

1. a) an immunoglobulin heavy chain variable domain (V) comprising heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2), and heavy chain complementarity determining region 3 (HCDR3) substantially similar to the amino acid sequences of SEQ ID NO:5, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17, respectively; Ｈ ) amino acid sequence; and b) an immunoglobulin light chain variable domain (V) comprising a LCDR1, a LCDR2, and a LCDR3 substantially similar to the light chain complementarity determining region 1 (LCDR1), the light chain complementarity determining region 2 (LCDR2), and the light chain complementarity determining region 3 (LCDR3), respectively, of the amino acid sequence of SEQ ID NO:21, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, or SEQ ID NO:30; Ｌ ) Amino acid sequence A polypeptide comprising: A polypeptide that is an antibody or an antigen-binding fragment thereof.

2. SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14) V selected from Ｈ and V Ｌ 2. The polypeptide of claim 1, comprising HCDR1, HCDR2 and HCDR3, and LCDR1, LCDR2 and LCDR3 of an antibody comprising the pair.

3. SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14) V selected from Ｈ and V Ｌ The polypeptide of claim 1 comprising a pair.

4. a) an immunoglobulin heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 92 or SEQ ID NO: 106; and / or b) an immunoglobulin light chain (LC) comprising the amino acid sequence of SEQ ID NO: 120; A polypeptide comprising: A polypeptide that is an antibody or an antigen-binding fragment thereof.

5. SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14), or A combination of the above an immunoglobulin heavy chain variable domain (V) comprising a paratope substantially similar to the paratope of an antibody comprising an amino acid sequence selected from Ｈ ) and an immunoglobulin light chain variable domain (V Ｌ and A polypeptide that is an antibody or an antigen-binding fragment thereof.

6. Immunoglobulin heavy chain variable domain (V Ｈ ) wherein the V Ｈ is the amino acid sequence of SEQ ID NO: 2, X １ is I or V; X ２ is W or S; X ３ is D or S; X ４ is S, T, Y or A; X ５ is N, Y, D or T; X ６ is K, T or I; X ７ is A, S or T; X ８ is A or S; X ９ is L, D, E or S; X １０ is V or I; X １１ is H, F or Y; X １２ is A or S; X １３ is F, M or L; X １４ is I or V, or any combination of the above) Including, and polypeptides that are antibodies or antigen-binding fragments thereof.

7. Immunoglobulin heavy chain variable domain (V Ｈ ) wherein the V Ｈ is the amino acid sequence of SEQ ID NO: 2, X １ is V; X ２ is S; X ３ is S; X ４ is T, Y or A; X ５ is Y, D or T; X ６ is T or I; X ７ is S or T; X ８ is S; X ９ is D, E or S; X １０ is I; X １１ is F or Y, X １２ is S; X １３ is M or L; X １４ is V, or any combination of the above) Including, and polypeptides that are antibodies or antigen-binding fragments thereof.

8. Immunoglobulin light chain variable domain (V Ｌ 8. The polypeptide of claim 6 or 7, further comprising: Ｌ comprises the amino acid sequence of SEQ ID NO: 18, and wherein: X １５ is N or Y; X １６ is L or I; X １７ is S or R; X １８ is K, F or Y; X １９ is S or N; X ２０ is V or I; X ２１ is W or Y; X ２２ is D, V or S; X ２３ is S, M or E; X ２４ is S, A or T; X ２５ is S, D or E; X ２６ is D, S, R or F; X ２７ is H, L, K or E, or any combination of the foregoing, polypeptides.

9. X １５ is Y; X １６ is I; X １７ is R; X １８ is F or Y; X １９ is N; X ２０ is I; X ２１ is Y; X ２２ is V or S; X ２３ is M or E; X ２４ is A or T; X ２５ is D or E; X ２６ is S, R or F; X ２７ is L, K or E, or any combination of the foregoing, a polypeptide according to claim 8.

10. The polypeptide according to any one of claims 1 to 9, which specifically binds to thymic stromal lymphopoietin (TSLP).

11. The antigen-binding fragment may comprise a single-chain fragment variable (scFv), a variable heavy domain of the heavy chain (V ＨＨ ), fragment antigen binding (Fab), Fab' or F(ab') ２ The polypeptide according to any one of claims 1 to 3 and 5 to 10, comprising:

12. The polypeptide of any one of claims 1 to 11, comprising an antibody heavy chain constant domain, an antibody light chain constant domain, or both an antibody heavy chain constant domain and an antibody light chain constant domain.

13. a) the antibody heavy chain constant domain is an IgG1, IgG2, IgG3, or IgG4 constant domain; b) the antibody light chain constant domain is a kappa constant domain or a lambda constant domain; or Both a) and b) are true; The polypeptide of claim 12.

14. 14. The polypeptide of claim 12 or 13, wherein the antibody heavy chain constant domain comprises one or more mutations that increase the serum half-life of the antibody or antigen-binding fragment thereof in humans, and optionally the antibody heavy chain constant domain comprises amino acid substitutions at amino acid residues 252, 254 and 256 (wherein the amino acid residues are numbered according to the EU index as in Kabat) with tyrosine, threonine and glutamic acid, respectively, compared to a wild-type human IgG constant domain.

15. The polypeptide of any one of claims 12 to 14, wherein the antibody heavy chain constant domain comprises the amino acid sequence of SEQ ID NO:

85.

16. A fusion protein comprising the polypeptide of any one of claims 1 to 15.

17. A polynucleotide comprising a nucleotide sequence encoding the polypeptide of any one of claims 1 to 15 or the fusion protein of claim 16.

18. A vector comprising the polynucleotide of claim 17.

19. 19. A host cell comprising the polynucleotide of claim 17 or the vector of claim 18.

20. A composition comprising a polypeptide according to any one of claims 1 to 15, a fusion protein according to claim 16, a polynucleotide according to claim 17, a vector according to claim 18, or a host cell according to claim 19.

21. 21. The composition of claim 20, further comprising one or more pharmaceutical excipients, diluents, or carriers.

22. 22. The composition of claim 20 or 21, further comprising one or more additional therapeutic agents.

23. 23. The composition of claim 22, wherein the one or more additional therapeutic agents comprise a corticosteroid, a beta agonist, a muscarinic antagonist, or a combination thereof.

24. 24. A method of treating a subject in need thereof, comprising administering to the subject an effective amount of the composition of any one of claims 20 to 23, optionally wherein the subject is a human.

25. 25. The method of claim 24, further comprising administering to the subject a therapeutically effective amount of an additional therapeutic or prophylactic agent.

26. 26. The method of claim 25, wherein the additional therapeutic or prophylactic agent comprises a corticosteroid, a beta agonist, a muscarinic antagonist, or a combination thereof.

27. 22. A method of reducing binding of thymic stromal lymphopoietin (TSLP) to TSLP receptor (TSLPR) on cells of a subject, comprising contacting the cells with an effective amount of the composition of any one of claims 19 to 21, optionally wherein the subject is a human.

28. 28. The method of any one of claims 24 to 27, wherein the subject is a pediatric patient, and optionally the subject is 12 years of age or older.

29. 29. The method of any one of claims 24 to 28, wherein the subject has asthma, atopic dermatitis (AD), allergic conjunctivitis, chronic obstructive pulmonary disease (COPD), chronic spontaneous urticaria (CSU), rheumatoid arthritis (RA), rhinosinusitis (RS), eosinophilic esophagitis (EE), or food hypersensitivity, and optionally, the subject has either COPD and / or moderate asthma or severe asthma.

30. 16. A method of making a polypeptide according to any one of claims 1 to 15, comprising culturing a host cell comprising a nucleotide sequence encoding said polypeptide under conditions whereby said polypeptide is expressed in said host cell.

31. 1. A computer-implemented method comprising: Scoring a polypeptide comprising an amino acid sequence with a computational binding optimization (CBO) model for functional properties related to modulating the activity of a target molecule. wherein the CBO model comprises: calculating, for each amino acid position of the amino acid sequence of the polypeptide, a plurality of energy scores based on the amino acid at a given position in the amino acid sequence; Add each calculated energy score to the array, generating a normalized sum of the energy scores for each position; and A method in which a score for the polypeptide is generated by summing each calculated energy score, and the score represents a functional property of the polypeptide related to the polypeptide regulating the activity of the target molecule.

32. Calculating the logarithm of each calculated energy score 32. The computer-implemented method of claim 31 further comprising: summing each calculated energy score is performed by summing the logarithm of each calculated energy score; The method, wherein the energy score is further calculated based on substituting the amino acid at a given position in the amino acid sequence with each of a plurality of different amino acids.

33. 33. The computer-implemented method of claim 31 or 32, wherein scoring the polypeptide using the CBO model is implementable by the script in Appendix C, and wherein the CBO model is substantially similar to the table in Appendix D.

34. The functional property is K below 10 pM Ｄ a binding affinity to a thymic stromal lymphopoietin (TSLP) polypeptide characterized by: (optionally as measured by KinExA); Binding specificity to the AB loop region and the C-terminal region of helix D of the TSLP polypeptide; Neutralizing activity against the TSLP polypeptide (optionally full-length human TSLP); or Inhibitory activity against TSLP-mediated signal transduction, or a combination of the above 34. The method according to any one of claims 31 to 33, wherein the method is at least one of:

35. 35. The method of claim 34, wherein the target molecule is thymic stromal lymphopoietin (TSLP) and the functional property of the polypeptide modulates the activity of the TSLP.

36. 1. A system comprising: a processor; and memory in which computer code instructions are stored wherein said processor and said memory, together with said computer code instructions, Scoring a polypeptide containing a certain amino acid sequence using the CBO model for functional properties related to modulating the activity of a target molecule and the CBO model is configured to: calculating, for each amino acid position of the amino acid sequence of the polypeptide, a plurality of energy scores based on the amino acid at a given position in the amino acid sequence; Add each calculated energy score to the array, generating a normalized sum of the energy scores for each position; and A system that generates a score for the amino acid sequence by summing each calculated energy score, the score representing a functional property of the polypeptide related to the polypeptide regulating the activity of the target molecule.

37. the instructions are further configured to cause the system to calculate a logarithm of each calculated energy score; summing each calculated energy score is performed by summing the logarithm of each calculated energy score; 37. The system of claim 36, wherein the energy score is further based on substituting the amino acid at a given position in the amino acid sequence with each of a plurality of different amino acids.

38. 38. The system of claim 36 or 37, wherein scoring the polypeptide using the CBO model is implemented by the script in Appendix C, and the CBO model is substantially similar to the table in Appendix D.

39. The functional property is K below 10 pM Ｄ a binding affinity to a thymic stromal lymphopoietin (TSLP) polypeptide characterized by: (optionally as measured by KinExA); Binding specificity to the AB loop region and the C-terminal region of helix D of the TSLP polypeptide; Neutralizing activity against the TSLP polypeptide (optionally full-length human TSLP); or Inhibitory activity against TSLP-mediated signal transduction, or a combination of the above The system according to any one of claims 36 to 38, wherein the system is at least one of:

40. 40. The method of claim 39, wherein the target molecule is thymic stromal lymphopoietin (TSLP) and the functional property of the polypeptide modulates the activity of the TSLP.

41. A polypeptide that specifically binds to thymic stromal lymphopoietin (TSLP), A polypeptide comprising an amino acid sequence that is assigned a score above a predetermined threshold by a computational bond optimization (CBO) model when the amino acid sequence of said polypeptide is scored.

42. the polypeptide is scoreable by a script in Computer Program Listing Appendix C, and the CBO model is calculated by a table substantially similar to Computer Program Listing Appendix D, and the polypeptide is assigned a score calculated using Computer Program Listing Appendix D; The polypeptide is assigned a score by a script substantially similar to Computer Program Listing Appendix C, wherein the script utilizes a CBO model substantially similar to the tables in Computer Program Listing Appendix D, and wherein the polypeptide is assigned a score of at least about -7.7, -7.0, -6.5, -6.0, -5.5, -5.0, -4.5, -4.0, -3.5, -3.

0. , 2.9, -2.8, -2.7, -2.6, -2.5, -2.4, -2.3, -2.2, -2.1, -2.0, -1.9, -1.8, -1.7, -1.6, -1.5, -1.4, -1.3, -1.2, -1.1, -1.0, 0.9, -0.8, -0.7, -0.6, -0.5, -0.4, -0.3, -0.2, and -0.

1.

43. K below 10 pM Ｄ a binding affinity to a thymic stromal lymphopoietin (TSLP) polypeptide characterized by: (optionally as measured by KinExA); Binding specificity to the AB loop region and the C-terminal region of helix D of the TSLP polypeptide; Neutralizing activity against the TSLP polypeptide (optionally full-length human TSLP); or Inhibitory activity against TSLP-mediated signal transduction, or a combination of the above 43. The polypeptide of claim 41 or 42, having one or more properties selected from:

44. The CBO model is SEQ ID NO: 4 and SEQ ID NO: 20 (AB-1); SEQ ID NO:5 and SEQ ID NO:21 (AB-2); SEQ ID NO:6 and SEQ ID NO:20 (AB-3); SEQ ID NO:7 and SEQ ID NO:22 (AB-4); SEQ ID NO:8 and SEQ ID NO:23 (AB-5); SEQ ID NO:9 and SEQ ID NO:24 (AB-6); SEQ ID NO: 10 and SEQ ID NO: 25 (AB-7); SEQ ID NO: 11 and SEQ ID NO: 20 (AB-8); SEQ ID NO: 12 and SEQ ID NO: 20 (AB-9); SEQ ID NO: 13 and SEQ ID NO: 26 (AB-10); SEQ ID NO: 14 and SEQ ID NO: 27 (AB-11); SEQ ID NO: 15 and SEQ ID NO: 28 (AB-12); SEQ ID NO: 16 and SEQ ID NO: 29 (AB-13); or SEQ ID NO: 17 and SEQ ID NO: 30 (AB-14) V selected from Ｈ and V Ｌ 44. The polypeptide of any one of claims 41 to 43, which outputs a score that is equal to or exceeds the score from the CBO model for one or more of the reference polypeptides that comprise the pair.

45. 45. The polypeptide of any one of claims 41 to 44, which does not comprise a heavy chain comprising SEQ ID NO: 81 and a light chain comprising SEQ ID NO:

82.

46. A polypeptide comprising: A polypeptide comprising an amino acid sequence that is assigned a score above a predetermined threshold by a computational bond optimization (CBO) model when the amino acid sequence of said polypeptide is scored.

47. A polypeptide that binds to human thymic stromal lymphopoietin (TSLP), generating a polypeptide sequence in the CBO model; The generated polypeptide sequence is then subjected to the CBO model. calculating a plurality of energy scores for each amino acid position of the polypeptide sequence, wherein the energy scores are based on substituting the amino acid at a given position in the polypeptide sequence with each of a plurality of different amino acids; adding each calculated energy score to an array; generating a normalized sum of the energy scores for each position; calculating the logarithm of each calculated energy score; and generating a score sequence for said polypeptide by summing the logarithms of each calculated energy score, said score representing a functional property of said polypeptide's ability to bind to human TSLP. To verify by A polypeptide designed by a method comprising:

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