Antigen binding molecules targeting thymic stromal lymphopoietin (TSLP)

Abstract

In various embodiments, the disclosure provides polypeptides (e.g., antibodies and antigen-binding fragments thereof) that specifically bind thymic stromal lymphopoietin (TSLP) (e.g., full-length human TSLP). In various embodiments, the 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 for treating TSLP-associated diseases or disorders.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of the following applications: U.S. Provisional Application No. 63 / 369,152, filed July 22, 2022; U.S. Provisional Application No. 63 / 369,158, filed July 22, 2022; U.S. Provisional Application No. 63 / 369,165, filed July 22, 2022; U.S. Provisional Application No. 63 / 369,175, filed July 22, 2022; U.S. Provisional Application No. 63 / 369,202, filed July 22, 2022; U.S. Provisional Application No. 63 / 369,203, filed July 22, 2022; U.S. Provisional Application No. 63 / 369,204, filed July 22, 2022; and U.S. Provisional Application No. 63 / 369,206, filed July 22, 2022.

[0003] This application claims priority to U.S. Application No. 18 / 356,891, filed July 21, 2023, and is a continuation-in-part of that application.

[0004] The entire teachings of the foregoing applications are incorporated herein by reference. Without limiting the foregoing, the sequence listings and codes of the corresponding Appendices A and B of the foregoing applications are incorporated by reference, for example, the files claim.txt and fit_model.txt of U.S. Application No. 18 / 356,891.

[0005] XML format materials are incorporated through reference.

[0006] This application incorporates by reference the sequence list contained in the following Extensible Markup Language (XML) format file:

[0007] a) File name: 57081061005_Sequence_Listing.xml; created on July 22, 2023, with a size of 212,394 bytes.

[0008] Materials in ASCII files are incorporated by reference.

[0009] This application incorporates by reference a list of computer programs contained in the following ASCII file, which is submitted hereto:

[0010] a) File name: claim.py; uploaded on July 22, 2023, available at https: / / zenodo.org / record / 8174726 / files / claim.py?download=1 or https: / / doi.org / 10.5281 / zenodo.8174726, select "claim.py", size 4,774 bytes. The entire teachings of the above file are incorporated herein by reference in their entirety.

[0011] b) File name: fit_model.etab; uploaded on July 22, 2023, available at https: / / zenodo.org / record / 8174726 / files / fit_model.etab?download=1 or https: / / doi.org / 10.5281 / zenodo.8174726. Select "fit_model.etab" for its size (2,888,059 bytes). The entire teachings of the above file are incorporated herein by reference in their entirety.

[0012] The entire teachings of the aforementioned documents are incorporated herein by reference in their entirety. The script in claim.py is intended to run in a Python environment located in the same folder as fit_model.etab to use the data from fit_model.etab. Background Art

[0013] Thymic stromal lymphopoietin (TSLP) is a cytokine involved in the pathogenesis of various inflammatory diseases. TSLP is released when the epithelium is exposed to "offenders" such as allergens, triggering a downstream inflammatory cascade. TSLP receptor (TSLPR) is expressed in a variety of cell types, including airway smooth muscle cells (ASMC), basophils, dendritic cells, eosinophils, type 2 innate lymphocytes (ILC2), 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 compelling evidence that TSLP expression dysregulation can lead to allergic diseases through the development of type 2 inflammatory responses (Ziegler et al., The biology of thymic stromal lymphopoietin (TSLP), Adv Pharmacol. 66:129-55 (2013)). Summary of the Invention

[0014] There is an urgent need for therapeutic agents to modulate (e.g., reduce or neutralize) the binding and / or activity of thymic stromal lymphopoietin (TSLP). This disclosure provides such agents.

[0015] The disclosures provided herein are based in part on the finding that the peptides disclosed herein specifically bind to TSLP and exhibit a variety of additional beneficial properties. Therefore, this disclosure generally relates to peptides (e.g., antibodies), compositions (e.g., pharmaceutical compositions), and methods for modulating (e.g., reducing or neutralizing) TSLP function and / or activity (e.g., in vivo).

[0016] In addition, this document also provides peptides (e.g., antibodies and antigen-binding fragments thereof) that specifically bind TSLP (e.g., full-length human TSLP) or variants thereof. In some embodiments, the peptides disclosed herein bind TSLP at the AB loop region and the C-terminal region of the helix D.

[0017] In some embodiments, the peptide regulates (e.g., reduces, for example, inhibits) the binding between TSLP and TSLPR (e.g., alone or with interleukin-7 receptor α (IL-7Rα) to form a heterodimeric complex). In some embodiments, the peptide regulates (e.g., reduces, for example, inhibits) TSLP-mediated signaling in vitro and / or in vivo.

[0018] This disclosure provides, among other things, polypeptides (e.g., antibodies or antigen-binding fragments thereof) that specifically bind to TSLP, wherein said polypeptides have one or more properties selected from the following:

[0019] Binding affinity for TSLP, characterized by K D 10 pM or lower (e.g., as measured by KinExA);

[0020] Binding specificity to the AB loop region and C terminal region of the helical D of TSLP;

[0021] Neutralizing activity against TSLPs (e.g., whole-length human TSLPs); or

[0022] Inhibitory activity against TSLP-mediated signal transduction

[0023] Or a combination thereof.

[0024] This disclosure also provides, among other things, polypeptides (e.g., antibodies or antigen-binding fragments thereof) comprising and containing variable domains of immunoglobulin heavy chains selected from V H ) / Immunoglobulin light chain variable domain (V L The complementary sites of the combined antibodies are basically similar:

[0025] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0026] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0027] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0028] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0029] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0030] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0031] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0032] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0033] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0034] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0035] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0036] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0037] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0038] SEQ ID NO:17 and SEQ ID NO:30 (AB-14), or

[0039] The aforementioned combination,

[0040] The polypeptide does not contain the heavy chain containing SEQ ID NO:81 or the light chain containing SEQ ID NO:82.

[0041] In some embodiments, the polypeptide comprises V selected from the following H / V L The complementary sites of the combined antibodies are basically similar:

[0042] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0043] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0044] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0045] SEQ ID NO:7 and SEQ ID NO:22(AB-4), or

[0046] The aforementioned combination,

[0047] The polypeptide does not contain the heavy chain containing SEQ ID NO:81 or the light chain containing SEQ ID NO:82.

[0048] This disclosure also provides, among other things, a polypeptide (e.g., an antibody or an antigen-binding fragment thereof) comprising:

[0049] V H The amino acid sequence includes heavy chain complementarity-determining regions 1 (HCDR1), 2 (HCDR2), and 3 (HCDR3) that are substantially similar to HCDR1, HCDR2, and HCDR3 of any of the amino acid sequences in SEQ ID NO:4-17, respectively; and

[0050] V L The amino acid sequence includes light chain complementarity-determining regions 1 (LCDR1), 2 (LCDR2), and 3 (LCDR3) that are substantially similar to the LCDR1, LCDR2, and LCDR3 sequences of any one of the amino acid sequences in SEQ ID NO:20-30, respectively.

[0051] The polypeptide does not contain the heavy chain containing SEQ ID NO:81 or the light chain containing SEQ ID NO:82.

[0052] In some embodiments, the polypeptide comprises:

[0053] V H Amino acid sequences comprising HCDR1, HCDR2, and HCDR3 that are substantially similar to the amino acid sequences of any one of SEQ ID NO:4-7; and

[0054] V L The amino acid sequences comprising LCDR1, LCDR2, and LCDR3 that are substantially similar to the amino acid sequences of any one of SEQ ID NO:20-22.

[0055] In some embodiments, the peptide comprises HCDR1, HCDR2, and HCDR3 of an antibody, as well as LCDR1, LCDR2, and LCDR3, wherein the antibody comprises V selected from the group consisting of V. H / V L combination:

[0056] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0057] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0058] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0059] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0060] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0061] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0062] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0063] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0064] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0065] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0066] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0067] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0068] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0069] SEQ ID NO:17 and SEQ ID NO:30(AB-14).

[0070] In some embodiments, the polypeptide further comprises V selected from the following H / V L The complementary sites of the combined antibodies have 100% sequence identity:

[0071] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0072] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0073] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0074] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0075] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0076] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0077] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0078] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0079] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0080] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0081] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0082] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0083] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0084] SEQ ID NO:17 and SEQ ID NO:30(AB-14).

[0085] In some embodiments, the peptide comprises HCDR1, HCDR2, and HCDR3 of an antibody, as well as LCDR1, LCDR2, and LCDR3, wherein the antibody comprises V selected from the group consisting of V. H / V L combination:

[0086] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0087] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0088] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0089] SEQ ID NO:7 and SEQ ID NO:22(AB-4).

[0090] In some embodiments, the polypeptide comprises V selected from the following: H / V LThe complementary sites of the combined antibodies have 100% sequence identity:

[0091] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0092] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0093] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0094] SEQ ID NO:7 and SEQ ID NO:22(AB-4).

[0095] This disclosure also provides, among other things, a polypeptide (e.g., an antibody or an antigen-binding fragment thereof) comprising V containing SEQ ID NO:2. H ,in:

[0096] X1 is not I;

[0097] X2 is not W;

[0098] X3 is not D;

[0099] X4 is not S;

[0100] X5 is not N;

[0101] X6 is not K;

[0102] X7 is not A;

[0103] X8 is not A;

[0104] X9 is not L;

[0105] X 10 Not V;

[0106] X 11 Not H,

[0107] X 12 Not A;

[0108] X 13 Not F;

[0109] X 14 Not I,

[0110] Or any combination thereof.

[0111] In some embodiments, the polypeptide comprises V containing SEQ ID NO:18 L ,in:

[0112] X 15Not N;

[0113] X 16 Not L;

[0114] X 17 Not S;

[0115] X 18 Not K;

[0116] X 19 Not S;

[0117] X 20 Not V;

[0118] X 21 Not W;

[0119] X 22 Not D;

[0120] X 23 Not S;

[0121] X 24 Not S;

[0122] X 25 Not S;

[0123] X 26 Not D;

[0124] X 27 Not H,

[0125] Or any combination thereof.

[0126] Furthermore, this disclosure also provides a polypeptide (e.g., an antibody or its antigen-binding fragment) comprising V having at least 70% sequence identity with SEQ ID NO:3. H The sequence, V, has at least 70% sequence identity with SEQ ID NO:19. L Sequence, or both, where the V H The sequence does not contain SEQ ID NO:3, this V L The sequence does not contain SEQ ID NO:19, or both.

[0127] This disclosure also provides, among other things, a polypeptide comprising:

[0128] a) HCDR 1 containing the amino acid sequence of SEQ ID NO:31, HCDR 2 containing the amino acid sequence of SEQ ID NO:35, and HCDR 3 containing the amino acid sequence of SEQ ID NO:50; and

[0129] b) LCDR 1 containing the amino acid sequence of SEQ ID NO:60, LCDR 2 containing the amino acid sequence of SEQ ID NO:67, and LCDR 3 containing the amino acid sequence of SEQ ID NO:71.

[0130] The polypeptide is an antibody or its antigen-binding fragment.

[0131] This disclosure also provides, among other things, a polypeptide comprising:

[0132] a) HCDR1 composed of the amino acid sequence of SEQ ID NO:31, HCDR2 composed of the amino acid sequence of SEQ ID NO:35, and HCDR3 composed of the amino acid sequence of SEQ ID NO:50; and

[0133] b) LCDR1 composed of the amino acid sequence of SEQ ID NO:60, LCDR2 composed of the amino acid sequence of SEQ ID NO:67, and LCDR3 composed of the amino acid sequence of SEQ ID NO:71.

[0134] The polypeptide is an antibody or its antigen-binding fragment.

[0135] In some embodiments, the polypeptide comprises:

[0136] a)V H It is humanized, containing the human frame region, or a combination thereof;

[0137] b)V L It is humanized, containing the human frame region, or a combination thereof, or

[0138] Both a) and b)

[0139] In some embodiments, the polypeptide comprises:

[0140] a)V H It contains the amino acid sequence of SEQ ID NO:5;

[0141] b)V L It contains the amino acid sequence of SEQ ID NO:21, or

[0142] Both a) and b)

[0143] In some embodiments, the antigen-binding fragment includes a single-chain variable fragment (scFv) and a heavy-chain variable restructure domain (V). HH ), antigen-binding fragment (Fab), Fab' or F(ab')2.

[0144] In some embodiments, the polypeptide comprises:

[0145] a) Antibody heavy chain constant domain;

[0146] b) Constant structural domain of antibody light chain

[0147] Or both a) and b).

[0148] In some embodiments, the antibody heavy chain constant domain is an IgG1, IgG2, IgG3, or IgG4 constant domain.

[0149] In some embodiments, the antibody heavy chain constant domain is an IgG1 or IgG2 constant domain.

[0150] In some embodiments, the antibody heavy chain constant domain contains one or more mutations that increase the serum half-life of the antibody or its antigen-binding fragment in humans. In some embodiments, relative to the wild-type human IgG constant domain, the antibody heavy chain constant domain contains amino acid substitutions at amino acid residues 252, 254, and 256, respectively, with tyrosine, threonine, and glutamic acid, wherein these amino acid residues are numbered according to the EU index in Kabat.

[0151] In some embodiments, the polypeptide comprises:

[0152] a) An antibody heavy chain (HC) containing the amino acid sequence of SEQ ID NO:92;

[0153] b) An antibody light chain (LC) containing the amino acid sequence of SEQ ID NO:120, or

[0154] Both a) and b)

[0155] In some embodiments, the polypeptide comprises:

[0156] a) An antibody heavy chain (HC) containing the amino acid sequence of SEQ ID NO:106;

[0157] b) An antibody light chain (LC) containing the amino acid sequence of SEQ ID NO:120, or

[0158] Both a) and b)

[0159] In some embodiments, the peptide specifically binds to TSLP.

[0160] In some embodiments, the polypeptide is a fusion protein.

[0161] In some embodiments, this disclosure provides a polynucleotide encoding the polypeptide disclosed herein, a vector containing such polynucleotide, and / or a host cell containing such polynucleotide and / or the vector.

[0162] In some embodiments, this disclosure provides compositions comprising the polypeptides disclosed herein (e.g., pharmaceutical compositions). 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, one or more additional therapeutic agents include corticosteroids, β-agonists, muscarinic antagonists, anti-inflammatory agents, IL-4 and / or IL-13 antagonists (e.g., antibodies or antigen-binding fragments thereof targeting IL-13, IL-4, or IL-4R) or combinations thereof.

[0163] This disclosure also provides methods for treating subjects in need (e.g., subjects with TSLP-related diseases or conditions), methods comprising administering to the subject an effective amount of the peptide disclosed herein and / or a composition comprising one or more of the peptides disclosed herein (e.g., a pharmaceutical composition).

[0164] This disclosure also provides a method for modulating (e.g., reducing) TSLP-mediated signaling in cells (e.g., cells in a subject), the method comprising contacting the cells with peptides disclosed herein and / or compositions comprising one or more peptides disclosed herein (e.g., pharmaceutical compositions).

[0165] In some embodiments, the subject has asthma, atopic dermatitis (AD), allergic conjunctivitis, chronic obstructive pulmonary disease (COPD), chronic spontaneous urticaria (CSU), rheumatoid arthritis (RA), sinusitis (RS), eosinophilic esophagitis (EE), or food allergy. 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 moderate or severe asthma.

[0166] This disclosure also provides, among other things, a method for reducing the binding of TSLP to TSLP receptors (TSLPRs) on subject cells, the method comprising contacting the cells with an effective amount of a composition comprising one or more polypeptides disclosed herein (e.g., a pharmaceutical composition).

[0167] This disclosure provides, among other things, a method for preparing the polypeptide disclosed herein, comprising culturing a host cell containing a nucleotide sequence encoding the polypeptide under conditions in which the polypeptide is expressed in a host cell.

[0168] This disclosure also provides, among other things, a computer-implemented method, which includes scoring a peptide comprising an amino acid sequence using a computational combination optimization (CBO) model. For each amino acid position in the amino acid sequence of the peptide, the CBO model calculates multiple energy scores based on replacing an amino acid at a given position in the amino acid sequence with each of a plurality of different amino acids. 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 peptide, representing the functional properties of the peptide, by summing the logarithms of each calculated energy score.

[0169] This disclosure also provides, among other things, a system including a processor and a memory storing computer code instructions thereon. The processor and memory with the computer code instructions are configured to cause the system to score a peptide sequence using a CBO model. For each amino acid position in the amino acid sequence of the peptide, the CBO model calculates multiple energy scores based on replacing an amino acid at a given position in the amino acid sequence with each of a plurality of different amino acids. 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 peptide, representing the functional properties of the peptide, by summing the logarithms of each calculated energy score.

[0170] This disclosure provides, among other things, a polypeptide that specifically binds to thymic stromal lymphopoietin (TSLP) and contains an amino acid sequence, which, after scoring the amino acid sequence of the polypeptide, is assigned a score above a predetermined threshold by calculating a binding optimization (CBO) model.

[0171] This disclosure also provides peptides that bind to human thymic stromal lymphopoietin (TSLP) and are designed using a method that includes generating peptide sequences using a CBO model. In some embodiments, the peptide is further designed by calculating multiple energy scores for each amino acid position in the peptide sequence, after validating the generated peptide sequence using a CBO model. The energy scores may be based on replacing an amino acid at a given position in the peptide sequence with each of a plurality of different amino acids. In some embodiments, the peptide is further validated by adding each calculated energy score to an array. In some embodiments, the peptide is further validated by generating a normalized sum of the energy scores at each position. In some embodiments, the peptide is further validated by calculating the logarithm of each calculated energy score. In some embodiments, the peptide is further validated by generating a score for the peptide sequence by summing the logarithms of each calculated energy score. This score represents a functional property of the peptide's ability to bind human TSLP. In some embodiments, the peptide sequence is validated if the peptide score is above a certain threshold. Attached Figure Description

[0172] This patent or application document contains at least one color drawing. Upon request and payment of the necessary fees, the official authority will provide a copy of this patent or application publication with one or more color drawings.

[0173] The above will become apparent from the following more specific description of exemplary embodiments, as shown in the accompanying drawings, wherein the same reference numerals refer to the same parts in all different views. These drawings are not necessarily drawn to scale, but rather emphasize the embodiments.

[0174] In the attached diagram, "reference" refers to the reference antibody.

[0175] Figure 1 The amino acid sequence of human thymic stromal lymphopoietin (TSLP) (SEQ ID NO:1) has been described. Epitope residues that the reference antibody binds to as disclosed herein are indicated by asterisks.

[0176] Figure 2A-2B The heavy chain variable domain (V) of the reference antibody and example antibodies AB-1 to AB-14 is depicted. H Amino acid sequence alignment. Heavy chain complementarity-determining region (HCDR) amino acid sequences identified by ImMunoGeneTics (IMGT) numbering are underlined. Bold letters indicate reference antibodies and variable residues in AB-1 through AB-14 (marked with "X" throughout this disclosure). n "Specified". "*" indicates a complementary residue. See also V in Table 1. HCommon sequence (SEQ ID NO:2). Complementary residues are defined as antibody residues in the reference antibody that are within 5 angstroms of the antigen when binding to human TSLP.

[0177] Figures 3A-3B The light chain variable domain (V) of the reference antibody and example antibodies AB-1 to AB-14 is depicted. L Amino acid sequence alignment. Heavy chain complementarity-determining region (LCDR) amino acid sequences identified by ImMunoGeneTics (IMGT) numbering are underlined. Bold letters indicate reference antibodies and variable residues in AB-1 through AB-14 (marked with "X" throughout this disclosure). n (Specified). See also V in Table 2. L Common sequence (SEQ ID NO:18).

[0178] Figures 4A-4B This shows a non-restricted example of an anti-TSLP antibody generated using direct enzyme-linked immunosorbent assay (ELISA) and its binding profile to human TSLP (hTSLP).

[0179] Figures 5A-5B The binding profile of a non-restricted example of an anti-TSLP antibody generated using direct ELISA with cynomolgus monkey TSLP (cynoTSLP) is shown.

[0180] Figures 6A-6B The binding profile of a non-restricted example of an anti-TSLP antibody generated using a blocking ELISA with hTSLP is shown.

[0181] Figures 7A-7B The binding profile of a non-restricted example of an anti-TSLP antibody generated using a blocking ELISA with cynoTSLP is shown.

[0182] Figures 8A-8C This demonstrates a non-restricted example of an anti-TSLP antibody that neutralizes the binding of hTSLP to the TSLP receptor (TSLPR). Ba / F3-STAT5-Luc cells and STAT5 reporter cells expressing hTSLPR and hIL-7Rα were used to assess the binding between hTSLP and TSLPR based on hTSLP-induced STAT5 activity. Figure 8A This shows TSLP-mediated STAT5 signaling as a function of hTSLP concentration. Figures 8B-8C This demonstrates the effect of anti-TSLP antibodies on TSLP-mediated STAT5 signaling.

[0183] Figures 9A-9BThis is a graph showing the blocking of TSLP binding to human bone marrow dendritic cells (mDCs) from peripheral blood by an exemplary anti-TSLP antibody. The curve shows the effect of the anti-TSLP antibody on TSLP-induced CCL17 expression in mDCs from a representative donor.

[0184] Figure 10A-10D This is a graph showing the binding spectra generated by direct ELISA for AB-2a, AB-2b, reference antibody, IgG1 isotype control, and IgG2 isotype control. Figure 10A The binding spectrum with hTSLP is shown. Figure 10B The binding spectrum with cynoTSLP is shown. Figure 10C The binding spectrum with mouse TSLP is shown. Figure 10D The binding spectrum with human IL-7 is shown.

[0185] Figure 11A-11B This is a graph showing the Ba / F3 assay results for AB-2a, AB-2b, reference antibody, IgG1 isotype control, and IgG2 isotype control. Figure 11A This displays the results of human TSLPR / IL-7RαBa / F3 assay. Figure 11B The results of TSLPR / IL-7RαBa / F3 assays in cynomolgus monkeys are shown.

[0186] Figure 12A This is a diagram showing the zero-order (1002), first-order (1004), and second-order (1006) models.

[0187] Figure 12B This is a flowchart illustrating an example of a process for determining a claimed sequence according to an embodiment of this disclosure.

[0188] Figure 13 A computer network or similar digital processing environment in which embodiments of the invention can be implemented is shown.

[0189] Figure 14 yes Figure 13 A diagram illustrating an exemplary internal structure of a computer (e.g., a client processor / device or a server computer) in a computer system. Detailed Implementation

[0190] An exemplary embodiment is described below.

[0191] Several aspects of this disclosure are described below with reference to examples for illustrative purposes only. It should be understood that numerous specific details, relationships, and methods are set forth to provide a full understanding of this disclosure. However, those skilled in the art will readily recognize that this disclosure can be practiced without one or more of these specific details, or with other methods, protocols, reagents, cell lines, and animals. This disclosure is not limited to the order of the described activities or events, as some activities may occur in a different order and / or simultaneously with other activities or events. Furthermore, not all described activities, steps, or events are required to implement the methods according to this disclosure. Many of the techniques and procedures described or mentioned herein are well understood by those skilled in the art and are typically employed using conventional methods.

[0192] definition

[0193] Unless otherwise defined, all terms, annotations, and other scientific terms or expressions 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 such definitions should not necessarily be construed as representing a material difference from the meaning commonly understood in the art. It should also be understood that terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant field and / or as otherwise defined herein.

[0194] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0195] When describing the elements disclosed herein, the articles “a / an,” “the,” and “said” are intended to indicate that there are one or more such elements. Furthermore, one or more elements may be the same or different. For example, unless the context clearly indicates otherwise, “polypeptide” includes a single polypeptide and two or more polypeptides.

[0196] Throughout this specification and the following claims, unless the context otherwise requires, the term "comprise" and variations such as "comprises" and "comprising" shall be understood to imply inclusion of, for example, the stated whole or step or group of whole or steps, but not to exclude any other whole or step or group of whole or steps. When used herein, the term "comprise" may be replaced by the terms "containing" or "comprising".

[0197] As used herein, the term "composed of" excludes any element, step, or component not specified in the elements of the claim. When used herein, the term "substantially composed of" does not exclude materials or steps that do not substantially affect the essential and novel features of the claim.

[0198] This document also provides corresponding embodiments for each embodiment characterized by the terms “comprising,” “containing,” “including,” or “having,” wherein these terms are replaced by the terms “consisting of” and / or “substantially consisting of.”

[0199] As used herein, the connecting term "and / or" between multiple listed elements is understood to encompass both individual and combined options. For example, in the case where two elements are connected by "and / or," the first option means that the first element applies while the second element does not. The second option means that the second element applies while the first element does not. The third option means that both the first and second elements apply. Any one of these options is understood to fall within this meaning and thus satisfies the requirement of the term "and / or" as used herein. The simultaneous application of more than one of these options is also understood to fall within this meaning and thus satisfies the requirement of the term "and / or".

[0200] It should be understood that for all numerical limits describing some parameters in this application, such as “about,” “at least,” “less than,” “less than,” and “greater than,” the description necessarily also includes any ranges defined by the values ​​referenced. Thus, for example, describing “at least 1, 2, 3, 4, or 5” also describes, in particular, the ranges 1-2, 1-3, 1-4, 1-5, 2-3, 2-4, 2-5, 3-4, 3-5, and 4-5, and so on.

[0201] When a list is presented, unless otherwise stated, it should be understood that each individual element of the list and each combination of the list is a separate embodiment. For example, a list of embodiments presented as “A, B, or C” should be interpreted as including embodiments “A”, “B”, “C”, “A or B”, “A or C”, “B or C”, or “A, B, or C”.

[0202] As used herein, the term “about” refers to an acceptable range of error for a given value, as determined by one of ordinary skill in the art. Typically, the acceptable range of error for a given value depends at least in part on how the value is measured or determined, such as limitations of the measurement system. For example, according to practice in the art, “about” may mean within an acceptable standard deviation. Alternatively, “about” may indicate a range of ±20%, for example, ±10%, ±5%, or ±1% of a given value. It should be understood that the term “about” may precede any given value specified herein, except for the specific value used in the examples. When “about” precedes a range, such as “90–99.9%”, the term “about” should be understood to apply to both given values ​​of that range, such that “about 90–99.9%” means about 90% repeating to about 99.9%.

[0203] As used herein, the term "peptide" refers to a polymer of at least two amino acids covalently linked by an amide bond, regardless of length or post-translational modifications (e.g., glycosylation or phosphorylation). Peptides may contain any suitable L- and / or D-amino acids, such as common α-amino acids (e.g., alanine, glycine, valine), non-α-amino acids (e.g., β-alanine, 4-aminobutyric acid, 6-aminohexanoic acid, sarcosine, statine), and uncommon amino acids (e.g., citrulline, homocitrulline, homoserine, leucine, valine, ornithine). The amino, carboxyl, and / or other functional groups on the peptide may 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 disclosed, for example, in Green and Wuts, "Protecting Groups in Organic Synthesis," John Wiley and Sons, 1991. The functional groups of the peptide can also be derivatized (e.g., alkylated) or labeled (e.g., with a detectable label, such as a fluorophore or hapten) using methods known in the art. If desired, the peptide may contain one or more modifications (e.g., amino acid linkers, acylation, acetylation, amidation, methylation, terminal modifiers (e.g., cyclization), N-methyl-α-amino substitution). Furthermore, the peptide may be an analogue of a known and / or naturally occurring peptide, such as a peptide analogue having one or more conserved amino acid residues substituted.

[0204] As used herein, a “polynucleotide” is defined as a plurality of nucleotides and / or nucleotide analogs linked together in a single molecule. In some embodiments, the polynucleotides disclosed herein comprise deoxyribonucleotides. In some embodiments, the polynucleotide comprises 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 a “U”, or vice versa), or polymers comprising purine and pyrimidine bases, or other natural, chemically or biochemically modified, non-natural, or derived nucleotide bases. The backbone of a polynucleotide may comprise sugar and phosphate groups, modified or substituted sugar or phosphate groups, polymers of synthetic subunits such as aminophosphates, or combinations thereof.

[0205] As used herein, the term "sequence identity" refers to the degree, expressed as a percentage, of how many identical residues are present at the same positions when sequences are aligned to achieve the maximum level of identity. For sequence alignment and comparison, typically one sequence is designated as the reference sequence, and the test sequence is compared to this reference sequence. Sequence identity between the reference and test sequences is expressed as the percentage of positions where the reference and test sequences share the same nucleotide or amino acid along the entire length of the reference sequence when aligned to achieve the maximum level of identity. For example, if the test sequence has the same nucleotide residues at 70% of the same positions along the entire length of the reference sequence when aligned to achieve the maximum level of identity, the two sequences are considered to have 70% sequence identity.

[0206] Those skilled in the art can readily perform sequence alignment for comparison using appropriate alignment methods or algorithms to achieve the maximum level of identity. In some cases, alignment may include introduced gaps to provide the maximum level of identity. Examples include the local homology algorithm of Smith and Waterman, Adv. Appl. Math. 2:482 (1981), the homology matching algorithm of Needleman and Wunsch, J. Mol. Biol. 48:443 (1970), the similarity search method of Pearson and 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, Wisconsin), and visual inspection (see generally Ausubel et al., Current Protocols in Molecular Biology).

[0207] When using sequence comparison algorithms, the test and reference sequences are input into the computer, with subsequence coordinates specified if necessary, and the sequence algorithm program parameters are also specified. Based on the specified program parameters, the sequence comparison algorithm then calculates the percentage of sequence identity of one or more test sequences relative to the reference sequence. A commonly used tool for determining the percentage of sequence identity is the Basic Local Alignment Search Tool for Proteins (BLASTP), available from the National Center for Biotechnology Information at the National Library of Medicine, National Institutes of Health (Altschul et al., 1990).

[0208] As used herein, the term “substantially similar” means a polypeptide disclosed herein that is substantially similar in amino acid sequence (e.g., having 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 residue sequence identity) and substantially retains one or more functional properties of the specific polypeptide disclosed herein (e.g., AB-1). In some embodiments, one or more functional properties are selected from, but are not limited to, substantially similar binding affinity, substantially similar binding specificity, substantially similar inhibitory activity, substantially similar neutralizing activity, and substantially similar self-association properties.

[0209] As used herein, “complementarity-determining region (CDR)” encompasses any CDR defined by methods recognized in the art for identifying CDR residues on antibodies. See, for example, Kabat, E.A. et al., (1991) Sequences of Proteins of Immunological Interest, 5th ed., U.S. Department of Health and Human Services, NIH Publication No. 91-3242; Chothia, E.A. et al., (1989) Nature 342:877; Chothia, C. et al., (1987) J. Mol. Biol. 196:901-917; Al-lazikani et al., (1997) J. Molec. Biol. 273:927-948; and Almagro, J. Mol. Recognit. 17:132-143 (2004). See also hgmp.mrc.ac.uk and bioinf.org.uk / abs. When using the same methodology to identify the CDRs of two antibodies, determine that the two antibodies have the same CDRs for each other in terms of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3.

[0210] The scope of the antibody's frame region and CDR can be identified using one of several suitable methods well known in the art, such as by Kabat definition, Chothia definition, AbM definition, and / or contact definition. Publicly available and / or commercially available tools for identifying framework and / or CDR regions include IgBlast (available from www.ncbi.nlm.nih.gov / igblast / ), Scaligner (available from drugdesigntech at www.scaligner.com / ), IMGT rules and / or tools (see, e.g., www.imgt.org / IMGTScientificChart / Nomenclature / IMGT-FRCDRdefinition.html, also accessible at www.imgt.org / ), Chothia specification assignments (accessible at www.bioinf.org.uk / abs / chothia.html), antigen receptor numbering and receptor classification (ANARCI, accessible at opig.stats.ox.ac.uk / webapps / newsabdab / sabpred / anarci / ), or Paratome web server (accessible at www.ofranlab.org / paratome / ), or Paratome web server (accessible at www.ofranlab.org / paratome / , see Vered Kunik et al., Nucleic Acids Research, Vol. 40, No. W1, July 1, 2012, pp. W521-W524.

[0211] For example, for AB-2, the amino acid sequences HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3: (1) contain 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, as determined by IMGT number; (2) contain 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, as determined by Kabat number; and (3) contain 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, as determined by Chothia number.

[0212] As used herein, the term "complementary site" refers to a group of amino acid residues in an antibody or its antigen-binding fragment that facilitates an epitope binding interaction with a target protein. The binding interaction can be a hydrogen bond, salt bridge, van der Waals interaction, ionic bond, or a combination thereof. The binding interaction can be direct or indirect, for example, through a coordinating intermediate molecule, such as an ion or water. In some embodiments, the residues of the complementary site comprise only residues that are part of a defined CDR. In other embodiments, the residues of the complementary site further comprise one or more residues that are not part of a defined CDR.

[0213] As used herein, the term "antibody mimic" refers to a polypeptide that mimics the ability of an antibody to bind to an antigen, but whose structure differs from that of a natural antibody. Examples of antibody mimics include, but are not limited to, adnectin, affibodies, affilins, affimers, affitins, alphabodies, anticoagulants, avimers, DARPins, fynomers, Kunitz domain peptides, monomers, nanobodies, and nanoclamps.

[0214] As used in this article, the term "K" D Also known as the "binding constant," "equilibrium dissociation constant," or "affinity constant," it is a measure of the degree of reversible association between two molecular species (e.g., antibody and target protein) and includes both actual binding affinity and apparent binding affinity. Binding affinity can be determined using methods known in the art, including, for example, by measuring surface plasmon resonances, such as using biolayer interferometry (Octet, ForteBio) or surface plasmon resonance (Biacore) systems and assays. A reference comparing various surface techniques used to measure 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 bindingkinetics, Analytical Biochemistry 508:78-96 (2016), the contents of which are incorporated herein by reference in their entirety.

[0215] The terms “subject” or “patient” refer to an animal (e.g., a mammal, such as a human) that is diagnosed or suspected of having a TSLP-related disease or condition (e.g., a disease or condition associated with dysregulation of TSLP expression, such as asthma or atopic dermatitis (AD)), or an animal at risk of developing such a condition. Diagnosis can be made by any method or technique known in the art. Those skilled in the art will understand that a subject to treatment according to this disclosure may have undergone standard testing or may have been identified as a person at risk (without testing) due to the presence of one or more risk factors associated with a disease or condition.

[0216] The phrase “pharmaceutically acceptable” means that, within reasonable medical judgment, the substance or composition modified by this phrase is suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and is proportionate to a reasonable benefit / risk ratio.

[0217] As used herein, a “pharmaceutically acceptable salt” means that which, within reasonable medical judgment, is suitable for contact with mammalian tissues without excessive toxicity, irritation, anaphylactic response, etc., and is proportionate to a reasonable benefit / risk ratio. Such pharmaceutically acceptable salts are well known in the art. For example, SMBerge et al. described pharmaceutically acceptable salts in detail in the J. Pharmaceutical Sciences, 1977, 66, 1-19, the relevant teachings of which are incorporated herein by reference in their entirety. Pharmaceutically acceptable salts of pharmaceutical preparations / compounds described herein include salts derived from suitable inorganic and organic acids and suitable inorganic and organic bases.

[0218] Examples of salts derived from suitable acids include those with amino groups formed from: inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid; or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid; or salts with amino groups formed by other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts derived from suitable acids include adipates, alginates, ascorbic acid salts, aspartate salts, benzenesulfonates, benzoates, hydrogen sulfates, borates, butyrates, camphorates, camphorsulfonates, cinnamates, citrates, cyclopentylpropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-2-hydroxyethylsulfonate, glyceryl phosphate, gluconate, glutarate, glycolate, hemisulfate, heptaate, hexanoate, hydroiodide, hydroxybenzoate, and 2-hydroxy-2-ethylsulfonate. Salts, hydroxymaleate, lacturonate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pyrate, pectin salt, persulfate, 2-phenoxybenzoate, phenylacetate, 3-phenylpropionate, phosphate, p-pentanoate, propionate, pyruvate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0219] Salts that can form mono-, di-, or tri-acids, and these salts can exist in hydrated, solvated, or substantially anhydrous forms.

[0220] Salts derived from suitable bases include those derived from inorganic bases such as alkali metals, alkaline earth metals, and ammonium bases, as well as those derived from aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine, and methylpyridine or N-acetylene. + Salts of ((C1-C4)alkyl)4. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, barium, etc. Other pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions (e.g., halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate).

[0221] "Pharmaceutically acceptable carrier" means a non-toxic carrier or excipient that does not impair the pharmacological activity of the drug to which it is formulated and is non-toxic when administered at 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, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, partially saturated mixtures of glycerides of the composition's 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 carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0222] As used herein, “to treat” or “treatment” means taking steps to deliver a therapy to a subject (e.g., a mammal) in need (e.g., by administering one or more therapeutic agents to the mammal). “To treat” or “treatment” includes suppressing a disease or condition (e.g., by slowing or stopping its progression or causing the disease or condition to subside) and relieving symptoms caused by a disease or condition.

[0223] The term “treatment” or “treatment” refers to the medical administration of a subject with the aim of improving, alleviating, stabilizing (i.e., not worsening), preventing, or curing a disease, pathological condition, or disorder—such as the specific indications exemplified herein. This term includes active treatment (treatment aimed at improving a disease, pathological condition, or disorder), causal treatment (treatment addressing the cause of the associated disease, pathological condition, or disorder), palliative treatment (treatment designed to relieve symptoms), preventative treatment (treatment aimed at minimizing or partially or completely suppressing the progression of the associated disease, pathological condition, or disorder); and supportive treatment (treatment used to complement another therapy). Treatment also includes reducing the severity of a disease or condition; preventing the spread of a disease or condition; delaying or slowing the progression of a disease or condition; alleviating or mitigating a disease or condition; and reducing (whether partially or completely), whether detectable or undetectable. Alleviating or mitigating a disease or condition means that the severity and / or adverse clinical manifestations of the disease, disorder, or condition are reduced and / or the time course of progression is slowed or prolonged compared to the severity or time course under no treatment. “Treatment” can also mean extended survival compared to the expected survival without treatment. Those who need treatment include those who already have a condition or disability, those who are susceptible to a condition or disability, or those for whom prevention of a condition or disability is necessary.

[0224] "Pharmaceutical composition" refers to a formulation of one or more therapeutic agents and a medium generally accepted in the art for delivering a bioactive agent to a subject (e.g., a human). In some embodiments, a pharmaceutical composition may comprise one or more pharmaceutically acceptable excipients, diluents, or carriers. In some embodiments, a pharmaceutical composition suitable for the methods disclosed herein further comprises one or more pharmaceutically acceptable carriers.

[0225] "Pharmaceutically acceptable carriers, diluents, or excipients" include any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the U.S. Food and Drug Administration for use in humans or livestock.

[0226] "Pharmaceutically acceptable carriers" refer to components in a pharmaceutical composition that are non-toxic to the subject, other than the active ingredient. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives. In some embodiments, a carrier may be a diluent, adjuvant, excipient, or mediator administered with a pharmaceutical agent (e.g., a polynucleotide). Such mediators may be liquids, such as water; and oils, including petroleum, animal, plant, or synthetic oils, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. For example, 0.4% saline and 0.3% glycine may be used. These solutions are sterile and generally free of particulate matter. They can be sterilized using conventional, well-known sterilization techniques (e.g., filtration). Compositions may include pharmaceutically acceptable excipients to approximate physiological conditions, such as pH adjusters and buffers, stabilizers, thickeners, lubricants, and colorants, as needed. The concentration of the drug in such pharmaceutical formulations can vary considerably, from less than about 0.5% to at least about 1%, or up to 15% or 20%, 25%, 30%, 35%, 40%, 45%, or 50% by weight. The concentration is selected primarily based on the desired dose, liquid volume, viscosity, etc., depending on the administration method. Suitable media and formulations (including other human proteins, such as human serum albumin) are described, for example, in Remington: The Science and Practice of Pharmacy, 21. st Edition [Remington: Pharmaceutical Science and Practice, 21st Edition], Troy, DB editor, Lipincott Williams and Wilkins, Philadelphia, PA, 2006, Part 5, Pharmaceutical Manufacturing, pp. 691-1092 (e.g., pp. 958-89).

[0227] Non-limiting examples of pharmaceutically acceptable carriers include physiologically compatible solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic agents and absorption delay agents, such as salts, buffers, antioxidants, sugars, aqueous or non-aqueous carriers, preservatives, wetting agents, surfactants or emulsifiers, or combinations thereof.

[0228] Non-limiting examples of buffer solutions are acetic acid, citric acid, formic acid, succinic acid, phosphoric acid, carbonic acid, malic acid, aspartic acid, histidine, boric acid, Tris buffer, HEPPSO, and HEPES.

[0229] Non-limiting examples of antioxidants include ascorbic acid, methionine, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, lecithin, citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, and tartaric acid.

[0230] Non-limiting examples of amino acids are histidine, isoleucine, methionine, glycine, arginine, lysine, L-leucine, trileucine, alanine, glutamic acid, L-threonine, and 2-aniline.

[0231] Non-limiting examples of surfactants are polysorbates (e.g., polysorbate-20 or polysorbate-80); poloxamer (e.g., poloxamer 188); Triton; sodium octyl glycoside; lauroyl-, myristyl-, linoleyl-, or stearoyl-sulfobetaine; lauroyl-, myristyl-, linoleyl-, or stearoyl-sarcosine; linoleyl, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palmitamidopropyl-, or isostearamidopropyl-betaine (e.g., lauroamidopropyl); myristamidopropyl-, palmitamidopropyl-, or isostearamidopropyl-dimethylamine; sodium methylcocoyl-taurate or disodium methyloleylenyltaurate; and MONAQUA. TM Series (Mona Industries, Inc., Paterson, New Jersey), polyethylene glycol, polypropylene glycol, and copolymers of ethylene glycol and propylene glycol (e.g., PLURONICS) TM (e.g., PF68).

[0232] Non-limiting examples of preservatives are phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrite, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride, alkyl (methyl, ethyl, propyl, butyl, etc.) esters of p-hydroxybenzoic acid, benzalkonium chloride, benzyl chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof.

[0233] Non-limiting examples of sugars are monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, and non-reducing sugars such as glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, erythritol, glycerol, arabinitol, xylitol, sorbitol, mannitol, melibiose, pinetriose, raffinose, manntriose, stachyose, maltose, lactulose, maltose, glucol, maltitol, lactitol, or isomaltulose.

[0234] Non-limiting examples of salts are 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, phosphorous acid, and non-toxic organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanes, hydroxyalkanes, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Base addition salts include those derived from: alkaline earth metals such as sodium, potassium, magnesium, calcium, etc., and non-toxic organic amines such as N,N'-dibenzylethylenediamine, N-methylglucosamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine, etc. In some embodiments, the salt is sodium chloride (NaCl).

[0235] The pharmaceutical agents (e.g., polynucleotides) described herein can be prepared according to standard procedures and administered at doses selected to reduce, prevent, or eliminate or slow or stop the progression of the treated condition (see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Panama (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 for a general description of the methods of administration of various pharmaceutical agents for human treatment).

[0236] As used herein, “to administer” or “to administer” means to provide a subject who requires treatment or prevention with the compound, composition or pharmaceutically acceptable salt thereof described herein. Administerment may be performed, for example, once, multiple times and / or over one or more extended periods. Administerment includes direct administration (including self-administration) and indirect administration (including prescribing a drug or instructing a subject to consume a drug). For example, as used herein, instructing a subject (e.g., a patient) to self-administer a drug (e.g., a medication) or to have another person administer a drug and / or to a patient (e.g., a physician) is administering a drug to a subject.

[0237] "Therapeutic effective dose," "effective dose," or "effective amount" refers to the amount that, at the necessary dose and for the necessary duration, is effective in achieving the desired therapeutic outcome (e.g., treatment, healing, suppression or improvement of a physiological response or condition). A complete therapeutic effect does not necessarily occur with the administration of a single dose, but may occur after a series of doses. Therefore, a therapeutic effective dose can be administered in one or more doses. Therapeutic effective doses can vary depending on factors such as the disease state of the mammal (e.g., a human patient), age, sex, and weight, the method of administration, and the ability of the therapeutic agent or combination of therapeutic agents to elicit the desired response.

[0238] The effective dose of the drug to be administered can be determined by a clinician of ordinary skill using the guidance provided herein and other methods known in the art. Relevant factors include the given drug, the drug formulation, the route of administration, the type of disease or disorder, the identity of the subject receiving treatment (e.g., age, sex, weight) or host, etc. For example, a suitable dose could 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 body weight / treatment. Determining the dose for a specific drug, subject, and disease is entirely within the capabilities of a person of ordinary skill in the art. Preferably, the dose does not cause or produces minimal adverse side effects.

[0239] The desired response or expected outcome includes effects at the cellular, tissue, or clinical levels. Therefore, the term "therapeuticly effective amount" or its synonyms depend on the context of its application. For example, in some embodiments, it is the amount of composition sufficient to achieve a therapeutic response compared to a response obtained without application of the composition. In other embodiments, it is the amount that produces a beneficial or desired outcome in a subject compared to a control. As defined herein, the therapeutically effective amount of the compositions disclosed herein (e.g., pharmaceutical compositions) can be readily determined by a person skilled in the art using conventional methods known in the art. Dosing regimens and routes of administration can be adjusted to provide an optimal therapeutic response.

[0240] Thymic stromal lymphopoietin (TSLP)

[0241] As used herein, TSLP includes wild-type TSLP protein (e.g., wild-type human TSLP protein or its homologs) and its truncated forms, mutant and engineered versions of full-length and truncated TSLP proteins, and modified forms of full-length and truncated TSLP proteins (e.g., post-translational modifications).

[0242] A non-limiting example of a human TSLP sequence is GenBank: AAK67490.1 (SEQ ID NO: 1).

[0243] MFPFALLYVLSVSFRKIFILQLVGLVLTYDFTNCDFEKIKAAYLSTISKDLITYMSGTKSTEFNNTVSCSNRPHCLTEIQSLTFNPTAGCASLAKEMFAMKTKAALAIWCPGYSETQINATQAMKKRRKRKVTTNKCLEQVSQLQGLWRRFNRPLLKQQ (SEQ ID NO: 1).

[0244] A non-restrictive example of a cynomolgus monkey TSLP sequence is GenBank: EHH54440.1 (SEQ ID NO:83).

[0245] MKSLGQSKKEEVSFRKFFIFQLVGLVLTYDFTNCDFQKIEADYLRTISKDLITYMSGTKSTDFNNTVSCSNRPHCLTEIQSLTFNPTPRCASLAKEMFARKTKATLALWCPGYSETQINATQAMKKRRKRKVTTNKCLEQVSQLLGLWRRFIRTLLKKQ (SEQ ID NO: 83).

[0246] In some embodiments, the peptide 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 peptide binds to a TSLP protein containing the amino acid sequence of SEQ ID NO:1. In some embodiments, the peptide binds to both a TSLP protein containing the amino acid sequence of SEQ ID NO:1 and a TSLP protein containing the amino acid sequence of SEQ ID NO:83.

[0247] In some embodiments, the peptide 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 with the wild-type TSLP protein, for example, having 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 with the 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 97-99.2% sequence identity with the wild-type TSLP protein.

[0248] In some embodiments, the peptide binds to a modified TSLP protein.

[0249] In some embodiments, the peptide can bind one or more epitope residues in a TSLP protein (e.g., full-length human TSLP), such as epitope residues 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the TSLP protein. In some embodiments, the peptide can bind one or more epitope residues selected from N64, N65, T66, V67, S68, C69, S70, N71, R72, H74, C75, E78, R150, F151, and R153 of SEQ ID NO:1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all 15 residues).

[0250] In some embodiments, the peptide binds to one or more epitope residues of the TSLP protein (e.g., one or more epitope residues in SEQ ID NO:1). In some embodiments, the peptide binds to one or more epitope residues selected from N64, N65, T66, V67, S68, C69, S70, N71, R72, H74, C75, E78, R150, F151, and R153 of SEQ ID NO:1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all 15 residues).

[0251] Comparison of peptides

[0252] As used herein, the term "comparative peptide" or "comparative polypeptide" refers to a polypeptide (e.g., an immunoglobulin molecule) that specifically binds to TSLP proteins and is not a polypeptide disclosed herein. The sequences of the comparative polypeptide and the polypeptide disclosed herein can be compared to illustrate structural differences between them (e.g., differences at one or more amino acid positions, such as amino acid substitutions). The polypeptide disclosed herein has more than one non-substantial difference (e.g., one or more substantial differences) compared to the comparative polypeptide, such that the polypeptide disclosed herein will, under controlled conditions, exhibit one or more of the following (i.e., one, two, or all three): function differently in a different manner to achieve different results compared to the comparative polypeptide. The comparative polypeptide may differ from the polypeptide disclosed herein by one or more amino acids, for example, in some embodiments, differing 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, the peptides are compared with the peptides 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 of amino acid identity percentage.

[0253] In some embodiments, the comparative peptide is an antibody comprising:

[0254] a) The heavy chain complementarity determination region 1 (HCDR1), heavy chain complementarity determination region 2 (HCDR2), and heavy chain complementarity determination region 3 (HCDR3) sequences of SEQ ID NO:31, SEQ ID NO:33, and SEQ ID NO:48, respectively;

[0255] b) Light chain complementarity determination region 1 (LCDR1), light chain complementarity determination region 2 (LCDR2), and light chain complementarity determination region 3 (LCDR3) sequences of SEQ ID NO:59, SEQ ID NO:67, and SEQ ID NO:69, respectively, or both a) and b).

[0256] In some embodiments, the comparative peptide is an antibody comprising:

[0257] a) The HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:31, SEQ ID NO:33, and SEQ ID NO:48, respectively; and

[0258] b) The LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:59, SEQ ID NO:67, and SEQ ID NO:69, respectively.

[0259] See Table 3 and Figure 2 for SEQ ID NO:31, 33 and 48. See Table 3 and Figure 3 for SEQ ID NO:59, 67 and 69.

[0260] In some embodiments, the comparative peptide is an antibody comprising:

[0261] a) The immunoglobulin heavy chain variable region (V) containing the amino acid sequence of SEQ ID NO:3 H ) structural domain;

[0262] b) The immunoglobulin light chain variable region (V) containing the amino acid sequence of SEQ ID NO:19 L ) structural domain, or

[0263] Both a) and b)

[0264] In some embodiments, the comparative peptide is an antibody comprising:

[0265] a) The immunoglobulin heavy chain variable region (V) containing the amino acid sequence of SEQ ID NO:3 H ) structural domain; and

[0266] b) The immunoglobulin light chain variable region (V) containing the amino acid sequence of SEQ ID NO:19 L ) structural domain.

[0267] See Table 1 and Figure 2 for SEQ ID NO:3. See Table 2 and Figure 3 for SEQ ID NO:19.

[0268] In some embodiments, the comparing peptide is an antibody referred to herein as a "reference antibody". The reference antibody comprises:

[0269] a) A heavy chain containing the amino acid sequence of SEQ ID NO:81; and

[0270] b) A light chain containing the amino acid sequence of SEQ ID NO:82.

[0271] See Table 4 for SEQ ID NO:81. See Table 5 for SEQ ID NO:82.

[0272] The reference antibody binds to and neutralizes TSLP.

[0273] peptides

[0274] In some embodiments, the peptide specifically binds to TSLP.

[0275] In some embodiments, a polypeptide is an antibody or an antigen-binding fragment thereof.

[0276] Variable structure domain

[0277] In some embodiments, the polypeptide (e.g., an antibody or its antigen-binding fragment) includes an immunoglobulin heavy chain variable region (V... H ), immunoglobulin light chain variable region (V L (or both). In some embodiments, the polypeptide comprises V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO:3. H V, which has less than 100% sequence identity with the amino acid sequence of SEQ ID NO:19 L , or both.

[0278] In some embodiments, the polypeptide comprises V H (e.g., mammal V) H For example, rodents (e.g., mice) V H Primates (e.g., humans) V H In some embodiments, the polypeptide comprises humanized V. H (For example, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% humanization), including human frame regions, or both.

[0279] In some embodiments, the polypeptide comprises V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO:3. H .

[0280] In some embodiments, the polypeptide comprises V having at least about 70% sequence identity with the amino acid sequence of SEQ ID NO:3. H In some embodiments, the polypeptide comprises V H The amino acid sequence of the polypeptide has 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 with the amino acid sequence of SEQ ID NO:3. In some embodiments, the polypeptide comprises V having at least about 85% sequence identity with the amino acid sequence of SEQ ID NO:3. H In some embodiments, the polypeptide comprises V having at least about 90% sequence identity with the amino acid sequence of SEQ ID NO:3. H .

[0281] In some embodiments, the polypeptide comprises a V with at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved substitution) relative to the amino acid sequence of SEQ ID NO:3.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 with about 1-10 amino acid substitutions relative to the amino acid sequence of SEQ ID NO:3. H In some embodiments, at least one amino acid substitution replaces only the heavy chain complementarity-determining region 1 (HCDR1), heavy chain complementarity-determining region 2 (HCDR2), and / or heavy chain complementarity-determining region 3 (HCDR3) residues of SEQ ID NO:3. In some embodiments, at least one amino acid substitution replaces only the non-CDR residues of SEQ ID NO:3 (e.g., within the frame region).

[0282] 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 value of 0 or greater in BLOSUM62.

[0283] In some embodiments, the amino acid substitution is a highly conserved substitution. The term "highly conserved amino acid substitution" or "highly conserved substitution" refers to an amino acid substitution having a value of at least 1 (e.g., at least 2) in BLOSUM62.

[0284] In some embodiments, the polypeptide comprises V having 100% sequence identity with the amino acid sequence of SEQ ID NO:3. H In some embodiments, the polypeptide comprises V H It contains the amino acid sequence of SEQ ID NO:3.

[0285] In some embodiments, the polypeptide comprises V having at least about 70% sequence identity with an amino acid sequence that is any one or more of the amino acids in SEQ ID NO:4-17. H Table 1 shows the sequences identified as SEQ ID NO:4-17, which correspond to human V. H Domains. In some embodiments, the polypeptide includes V HThe amino acid sequence of the polypeptide has 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 with any one or more of the amino acid sequences in SEQ ID NO:4-17. In some embodiments, the polypeptide comprises V having at least about 85% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:4-17. H In some embodiments, the polypeptide comprises V having at least about 90% sequence identity with an amino acid sequence of any one or more of SEQ ID NO:4-17. H .

[0286] In some embodiments, the polypeptide comprises V H The polypeptide comprises at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:4-17. For example, the number of amino acid substitutions may 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 V H It contains about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO:4-17.

[0287] In some embodiments, the polypeptide comprises V having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:4-17. H In some embodiments, the polypeptide comprises V H It contains the amino acid sequence of any one of SEQ ID NO:4-17.

[0288] In some embodiments, the polypeptide comprises V having at least about 70% sequence identity with an amino acid sequence of any one or more of SEQ ID NO:4-7. H In some embodiments, the polypeptide comprises V HThe amino acid sequence of the polypeptide has 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 with any one or more of the amino acid sequences in SEQ ID NO:4-7. In some embodiments, the polypeptide comprises V having at least about 85% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:4-7. H In some embodiments, the polypeptide comprises V having at least about 90% sequence identity with an amino acid sequence of any one or more of SEQ ID NO:4-7. H .

[0289] In some embodiments, the polypeptide comprises V H The polypeptide comprises at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:4-7. For example, the number of amino acid substitutions may 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 V H It contains about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO:4-7.

[0290] In some embodiments, the polypeptide comprises V having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:4-7. H In some embodiments, the polypeptide comprises V H It contains the amino acid sequence of any one of SEQ ID NO:4-7.

[0291] In some embodiments, the polypeptide comprises V L (e.g., mammal V) L For example, rodents (e.g., mice) V L Primates (e.g., humans) V L In some embodiments, the polypeptide comprises humanized V. L (For example, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% humanization), including human frame regions, or both.

[0292] In some embodiments, the polypeptide comprises V having less than 100% sequence identity with the amino acid sequence of SEQ ID NO:19. L .

[0293] In some embodiments, the polypeptide comprises V having at least about 70% sequence identity with the amino acid sequence of SEQ ID NO:19. L In some embodiments, the polypeptide comprises V L The amino acid sequence of the polypeptide has 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 with the amino acid sequence of SEQ ID NO:19. In some embodiments, the polypeptide comprises V having at least about 85% sequence identity with the amino acid sequence of SEQ ID NO:19. L In some embodiments, the polypeptide comprises V having at least about 90% sequence identity with the amino acid sequence of SEQ ID NO:19. L .

[0294] In some embodiments, the polypeptide comprises a V with at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved substitution) relative to the amino acid sequence of SEQ ID NO:19. 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 with about 1-10 amino acid substitutions relative to the amino acid sequence of SEQ ID NO:19. L In some embodiments, at least one amino acid substitution replaces only light chain complementarity-determining region 1 (LCDR1), light chain complementarity-determining region 2 (LCDR2), and / or light chain complementarity-determining region 3 (LCDR3) residues of SEQ ID NO:19. In some embodiments, at least one amino acid substitution replaces only non-CDR residues of SEQ ID NO:19 (e.g., within the frame region).

[0295] In some embodiments, the amino acid substitution is a conservative substitution.

[0296] In some embodiments, the amino acid substitution is a highly conserved substitution.

[0297] In some embodiments, the polypeptide comprises V, which has 100% sequence identity with the amino acid sequence of SEQ ID NO:19. L In some embodiments, the polypeptide comprises V L It contains the amino acid sequence of SEQ ID NO:19.

[0298] In some embodiments, the polypeptide comprises V having at least about 70% sequence identity with an amino acid sequence that is any one or more of the amino acids in SEQ ID NO:20-30. L Table 2 shows the sequences identified as SEQ ID NO:20-30, which correspond to human V. L Domains. In some embodiments, the polypeptide includes V L The amino acid sequence of the polypeptide has 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 with any one or more of the amino acid sequences in SEQ ID NO:20-30. In some embodiments, the polypeptide comprises V having at least about 85% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:20-30. L In some embodiments, the polypeptide comprises V having at least about 90% sequence identity with an amino acid sequence of any one or more of SEQ ID NO:20-30. L .

[0299] In some embodiments, the polypeptide comprises V L The polypeptide comprises at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO: 20-30. For example, the number of amino acid substitutions may 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 V L It contains about 1-10 amino acid substitutions relative to the amino acid sequence of any one or more of SEQ ID NO:20-30.

[0300] In some embodiments, the polypeptide comprises V having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:20-30. L In some embodiments, the polypeptide comprises V L It contains the amino acid sequence of any one of SEQ ID NO:20-30.

[0301] In some embodiments, the polypeptide comprises V having at least about 70% sequence identity with an amino acid sequence that is any one or more of the amino acids in SEQ ID NO:20-22. L In some embodiments, the polypeptide comprises V L The amino acid sequence of the polypeptide has 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 with any one or more of the amino acid sequences in SEQ ID NO:20-22. In some embodiments, the polypeptide comprises V having at least about 85% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:20-22. L In some embodiments, the polypeptide comprises V having at least about 90% sequence identity with an amino acid sequence of any one or more of SEQ ID NO:20-22. L .

[0302] In some embodiments, the polypeptide comprises V L The polypeptide comprises at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:20-22. For example, the number of amino acid substitutions may 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 V L It contains about 1-10 amino acid substitutions relative to the amino acid sequence of any one or more of SEQ ID NO:20-22.

[0303] In some embodiments, the polypeptide comprises V having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:20-22. L In some embodiments, the polypeptide comprises V LIt contains the amino acid sequence of any one of SEQ ID NO:20-22.

[0304] In some embodiments, the polypeptide comprises V H (e.g., mammal V) H For example, rodents (e.g., mice) V H Primates (e.g., humans) V H ) and V L (e.g., mammal V) L For example, rodents (e.g., mice) V L Primates (e.g., humans) V L In some embodiments, the polypeptide comprises humanized V. H and V L (For example, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% humanization), including human frame regions, or both.

[0305] In some embodiments, the polypeptide comprises:

[0306] a)V H Its amino acid sequence has less than 100% sequence identity with that of SEQ ID NO:3;

[0307] b)V L Its amino acid sequence has less than 100% sequence identity with that of SEQ ID NO:19, or

[0308] Both a) and b)

[0309] In some embodiments, the polypeptide comprises:

[0310] a)V H Its amino acid sequence has less than 100% sequence identity with that of SEQ ID NO:3; and

[0311] b)V L Its amino acid sequence has less than 100% sequence identity with that of SEQ ID NO:19.

[0312] In some embodiments, the polypeptide comprises:

[0313] a)V H It has 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;

[0314] b)V LIts amino acid sequence has 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:19, or

[0315] Both a) and b)

[0316] The polypeptide does not contain V containing SEQ ID NO:3. H Amino acid sequence and V of SEQ ID NO:19 L The antibody's amino acid sequence contains all six CDRs.

[0317] In some embodiments, the polypeptide comprises:

[0318] a)V H It has 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; and

[0319] b)V L It has 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:19.

[0320] The polypeptide does not contain V containing SEQ ID NO:3. H Amino acid sequence and V of SEQ ID NO:19 L The antibody's amino acid sequence contains all six CDRs.

[0321] In some embodiments, the polypeptide comprises:

[0322] a)V H Its amino acid sequence relative to SEQ ID NO:3 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved substitution);

[0323] b)V L Its amino acid sequence relative to SEQ ID NO:19 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved substitution), or

[0324] Both a) and b)

[0325] In some embodiments, the polypeptide comprises:

[0326] a)V HIts amino acid sequence relative to SEQ ID NO:3 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution); and

[0327] b)V L Its amino acid sequence relative to SEQ ID NO:19 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution).

[0328] In some embodiments, the polypeptide comprises:

[0329] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution such as a highly conserved amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residues of SEQ ID NO:3;

[0330] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of the LCDR1, LCDR2 and / or LCDR3 residues of SEQ ID NO:19, or

[0331] Both a) and b)

[0332] In some embodiments, the polypeptide comprises:

[0333] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residues of SEQ ID NO:3; and

[0334] b)V L It contains at least one amino acid substitution of the LCDR1, LCDR2 and / or LCDR3 residues of SEQ ID NO:19 (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution).

[0335] In some embodiments, the polypeptide comprises:

[0336] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue of SEQ ID NO:3 (e.g., within the framework region);

[0337] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue (e.g., within the framework region) of SEQ ID NO:19, or

[0338] Both a) and b)

[0339] In some embodiments, the polypeptide comprises:

[0340] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue (e.g., within the framework region) of SEQ ID NO:3; and

[0341] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue (e.g., within the framework region) of SEQ ID NO:19.

[0342] In some embodiments, the polypeptide comprises:

[0343] a)V H It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:4-17;

[0344] b)V L It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:20-30, or both a) and b).

[0345] In some embodiments, the polypeptide comprises:

[0346] a)V H It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:4-17; and

[0347] b)V L It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:20-30.

[0348] In some embodiments, the polypeptide comprises:

[0349] a)V HIt contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) relative to any one or more amino acid sequences in SEQ ID NO:4-17;

[0350] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) relative to any one or more amino acid sequences in SEQ ID NO:20-30, or

[0351] Both a) and b)

[0352] In some embodiments, the polypeptide comprises:

[0353] a)V H It contains at least one amino acid substitution relative to any one or more amino acid sequences in SEQ ID NO:4-17 (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution); and

[0354] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) relative to any one or more amino acid sequences in SEQ ID NO:20-30.

[0355] In some embodiments, the polypeptide comprises:

[0356] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution such as a highly conserved amino acid substitution) of any one or more HCDR1, HCDR2 and / or HCDR3 residues in SEQ ID NO:4-17;

[0357] b)V L It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more of the LCDR1, LCDR2 and / or LCDR3 residues in SEQ ID NO:20-30, or

[0358] Both a) and b)

[0359] In some embodiments, the polypeptide comprises:

[0360] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more HCDR1, HCDR2 and / or HCDR3 residues in SEQ ID NO:4-17; and

[0361] b)V LIt contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more of the LCDR1, LCDR2 and / or LCDR3 residues in SEQ ID NO:20-30.

[0362] In some embodiments, the polypeptide comprises:

[0363] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) in SEQ ID NO:4-17;

[0364] b)V L It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) of SEQ ID NO:20-30, or

[0365] Both a) and b)

[0366] In some embodiments, the polypeptide comprises:

[0367] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) of SEQ ID NO:4-17; and

[0368] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) in SEQ ID NO:20-30.

[0369] In some embodiments, the polypeptide disclosed herein comprises:

[0370] a)V H It contains the amino acid sequence of any one of SEQ ID NO:4-17; and

[0371] b)V L It contains the amino acid sequence of any one of SEQ ID NO:20-30.

[0372] In some embodiments, the polypeptide comprises:

[0373] a)V H It contains the amino acid sequence of any one of SEQ ID NO:4-17; and

[0374] b)V L It contains the amino acid sequence of SEQ ID NO:19.

[0375] In some embodiments, the polypeptide comprises:

[0376] a)V H It contains the amino acid sequence of SEQ ID NO:3; and

[0377] b)V L It contains the amino acid sequence of any one of SEQ ID NO:20-30.

[0378] In some embodiments, the polypeptide comprises:

[0379] a)V H It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:4-7;

[0380] b)V L It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:20-22, or both a) and b).

[0381] In some embodiments, the polypeptide comprises:

[0382] a)V H It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:4-7; and

[0383] b)V L It has at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, about 95%, 98%, or 99%) sequence identity with any one or more of the amino acid sequences in SEQ ID NO:20-22.

[0384] In some embodiments, the polypeptide comprises:

[0385] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) relative to any one or more amino acid sequences in SEQ ID NO:4-7;

[0386] b)V LIt contains at least one amino acid substitution relative to any one or more amino acid sequences in SEQ ID NO:20-22 (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution), or

[0387] Both a) and b)

[0388] In some embodiments, the polypeptide comprises:

[0389] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) relative to any one or more amino acid sequences in SEQ ID NO:4-7; and

[0390] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) relative to any one or more amino acid sequences in SEQ ID NO:20-22.

[0391] In some embodiments, the polypeptide comprises:

[0392] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more HCDR1, HCDR2 and / or HCDR3 residues in SEQ ID NO:4-7;

[0393] b)V L It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more of the LCDR1, LCDR2 and / or LCDR3 residues in SEQ ID NO:20-22, or

[0394] Both a) and b)

[0395] In some embodiments, the polypeptide comprises:

[0396] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more HCDR1, HCDR2 and / or HCDR3 residues in SEQ ID NO:4-7; and

[0397] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more of the LCDR1, LCDR2 and / or LCDR3 residues in SEQ ID NO:20-22.

[0398] In some embodiments, the polypeptide comprises:

[0399] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) in SEQ ID NO:4-7;

[0400] b)V L It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) of SEQ ID NO:20-22, or

[0401] Both a) and b)

[0402] In some embodiments, the polypeptide comprises:

[0403] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) of SEQ ID NO:4-7; and

[0404] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of any one or more non-CDR residues (e.g., within the framework region) in SEQ ID NO:20-22.

[0405] In some embodiments, the polypeptide disclosed herein comprises:

[0406] a)V H It contains the amino acid sequence of any one of SEQ ID NO:4-7; and

[0407] b)V L It contains the amino acid sequence of any one of SEQ ID NO:20-22.

[0408] In some embodiments, the polypeptide comprises:

[0409] a)V H It contains the amino acid sequence of any one of SEQ ID NO:4-7; and

[0410] b)V L It contains the amino acid sequence of SEQ ID NO:19.

[0411] In some embodiments, the polypeptide comprises:

[0412] a)VH It contains the amino acid sequence of SEQ ID NO:3; and

[0413] b)V L It contains the amino acid sequence of any one of SEQ ID NO:20-22.

[0414] In some embodiments, the polypeptide comprises:

[0415] a)V H It has 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;

[0416] b)V L Its amino acid sequence has 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, or

[0417] Both a) and b)

[0418] In some embodiments, the polypeptide comprises:

[0419] a)V H It has 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; and

[0420] b)V L It has 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.

[0421] In some embodiments, the polypeptide comprises:

[0422] a)V H Its amino acid sequence relative to SEQ ID NO:5 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution);

[0423] b)V L Its amino acid sequence relative to SEQ ID NO:21 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution), or

[0424] Both a) and b)

[0425] In some embodiments, the polypeptide comprises:

[0426] a)V H Its amino acid sequence relative to SEQ ID NO:5 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution); and

[0427] b)V L Its amino acid sequence relative to SEQ ID NO:21 contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution).

[0428] In some embodiments, the polypeptide comprises:

[0429] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution such as a highly conserved amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residues of SEQ ID NO:5;

[0430] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of the LCDR1, LCDR2 and / or LCDR3 residues of SEQ ID NO:21, or

[0431] Both a) and b)

[0432] In some embodiments, the polypeptide comprises:

[0433] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of the HCDR1, HCDR2 and / or HCDR3 residues of SEQ ID NO:5; and

[0434] b)V L It contains at least one amino acid substitution of the LCDR1, LCDR2 and / or LCDR3 residues of SEQ ID NO:21 (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution).

[0435] In some embodiments, the polypeptide comprises:

[0436] a)V H It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue (e.g., within the framework region) of SEQ ID NO:5;

[0437] b)V LIt contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue of SEQ ID NO:21 (e.g., within the framework region), or

[0438] Both a) and b)

[0439] In some embodiments, the polypeptide comprises:

[0440] a)V H It comprises at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue (e.g., within the framework region) of SEQ ID NO:5; and

[0441] b)V L It contains at least one amino acid substitution (e.g., at least one conserved substitution, such as a highly conserved amino acid substitution) of a non-CDR residue of SEQ ID NO:21 (e.g., within the framework region).

[0442] In some embodiments, the polypeptide disclosed herein comprises:

[0443] a)V H It contains the amino acid sequence of SEQ ID NO:4, and

[0444] b)V L It contains the amino acid sequence of SEQ ID NO:20(AB-1). In some embodiments, the polypeptide disclosed herein comprises:

[0445] a)V H It contains the amino acid sequence of SEQ ID NO:5, and

[0446] b)V L It contains the amino acid sequence of SEQ ID NO:21(AB-2). In some embodiments, the polypeptide disclosed herein comprises:

[0447] a)V H It contains the amino acid sequence of SEQ ID NO:6, and

[0448] b)V L It contains the amino acid sequence of SEQ ID NO:20(AB-3). In some embodiments, the polypeptide disclosed herein comprises:

[0449] a)V H It contains the amino acid sequence of SEQ ID NO:7, and

[0450] b)V LIt contains the amino acid sequence of SEQ ID NO:22(AB-4). In some embodiments, the polypeptide disclosed herein comprises:

[0451] a)V H It contains the amino acid sequence of SEQ ID NO:8, and

[0452] b)V L It contains the amino acid sequence of SEQ ID NO:23 (AB-5). In some embodiments, the polypeptide disclosed herein comprises:

[0453] a)V H It contains the amino acid sequence of SEQ ID NO:9, and

[0454] b)V L It contains the amino acid sequence of SEQ ID NO:24 (AB-6). In some embodiments, the polypeptide disclosed herein comprises:

[0455] a)V H It contains the amino acid sequence of SEQ ID NO:10, and

[0456] b)V L It contains the amino acid sequence of SEQ ID NO:25 (AB-7). In some embodiments, the polypeptide disclosed herein comprises:

[0457] a)V H It contains the amino acid sequence of SEQ ID NO:11, and

[0458] b)V L It contains the amino acid sequence of SEQ ID NO:20 (AB-8). In some embodiments, the polypeptide disclosed herein comprises:

[0459] a)V H It contains the amino acid sequence of SEQ ID NO:12, and

[0460] b)V L It contains the amino acid sequence of SEQ ID NO:20 (AB-9). In some embodiments, the polypeptide disclosed herein comprises:

[0461] a)V H It contains the amino acid sequence of SEQ ID NO:13, and

[0462] b)V L It contains the amino acid sequence of SEQ ID NO:26(AB-10).

[0463] In some embodiments, the polypeptide disclosed herein comprises:

[0464] a)V H It contains the amino acid sequence of SEQ ID NO:14, and

[0465] b)V L It contains the amino acid sequence of SEQ ID NO:27(AB-11).

[0466] In some embodiments, the polypeptide disclosed herein comprises:

[0467] a)V H It contains the amino acid sequence of SEQ ID NO:15, and

[0468] b)V L It contains the amino acid sequence of SEQ ID NO:28(AB-12).

[0469] In some embodiments, the polypeptide disclosed herein comprises:

[0470] a)V H It contains the amino acid sequence of SEQ ID NO:16, and

[0471] b)V L It contains the amino acid sequence of SEQ ID NO:29(AB-13).

[0472] In some embodiments, the polypeptide disclosed herein comprises:

[0473] a)V H It contains the amino acid sequence of SEQ ID NO:17, and

[0474] b)V L It contains the amino acid sequence of SEQ ID NO:30(AB-14).

[0475] Complementarity Determinant Region (CDR)

[0476] CDRs (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and / or LCDR3) can be defined by any method recognized in the art for identifying CDR residues of antibodies, as further described herein (e.g., CDRs defined by Kabat, CDRs defined by Chothia, or CDRs defined by IMGT).

[0477] In some embodiments, the polypeptide (e.g., an antibody or its antigen-binding fragment) does not contain V containing SEQ ID NO:3. H Amino acid sequence and V of SEQ ID NO:19L The antibody contains all six CDRs of the amino acid sequence. In some embodiments, the polypeptide does not contain all six sequences of 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 contain all four sequences of SEQ ID NO:33, SEQ ID NO:48, SEQ ID NO:59, and SEQ ID NO:69.

[0478] In some embodiments, the polypeptide comprises fewer than six (e.g., 1, 2, 3, 4, or 5) CDRs of an antibody, the antibody comprising the V of SEQ ID NO:3. H Amino acid sequence and V of SEQ ID NO:19 L Amino acid sequence. In some embodiments, the polypeptide comprises 1, 2, 3, 4, or 5 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, but not all 6 sequences. In some embodiments, the polypeptide comprises 1, 2, or 3 sequences selected from SEQ ID NO:33, SEQ ID NO:48, SEQ ID NO:59, and SEQ ID NO:69, but not all 4 sequences.

[0479] In some embodiments, the polypeptide comprises all six CDRs of the specific polypeptide disclosed herein. In some embodiments, the polypeptide comprises fewer than six (e.g., 1, 2, 3, 4, or 5) CDRs of the specific polypeptide disclosed herein.

[0480] In some embodiments, the polypeptide comprises:

[0481] a)V H The amino acid sequence contains V that is respectively associated with one or more of the sequences in SEQ ID NO:4-17. H HCDR1, HCDR2 and HCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity);

[0482] b)V L The amino acid sequence contains V that is respectively associated with one or more of SEQ ID NO:20-30. LLCDR1, LCDR2, and LCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity), or

[0483] Both a) and b)

[0484] In some embodiments, the polypeptide comprises:

[0485] a)V H The amino acid sequence contains V that is respectively associated with one or more of the sequences in SEQ ID NO:4-17. H HCDR1, HCDR2, and HCDR3 with fundamentally similar amino acid sequences (e.g., having at least about 90% sequence identity); and

[0486] b)V L The amino acid sequence contains V that is respectively associated with one or more of SEQ ID NO:20-30. L LCDR1, LCDR2, and LCDR3 have fundamentally similar amino acid sequences (e.g., having at least about 90% sequence identity).

[0487] See Table 2-3 and Figures 2A-3B Non-limiting examples of SEQ ID NOs:4-17 and 20-30, and the corresponding HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences.

[0488] In some embodiments, the polypeptide comprises:

[0489] a)V H The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:4-17 respectively. H The amino acid sequence of HCDR1, HCDR2, and HCDR3 has one or more functional characteristics of HCDR1, HCDR2, and HCDR3; or

[0490] b)V L The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:20-30 respectively. L The amino acid sequence of LCDR1, LCDR2, and LCDR3 contains one or more functional characteristics of LCDR1, LCDR2, and LCDR3, or

[0491] Both a) and b)

[0492] In some embodiments, the polypeptide comprises:

[0493] a)V H The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:4-17 respectively. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain one or more functional characteristics of HCDR1, HCDR2, and HCDR3; and

[0494] b)V L The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:20-30 respectively. L One or more functional properties of the amino acid sequences of LCDR1, LCDR2 and LCDR3.

[0495] In some embodiments, the polypeptide comprises:

[0496] a)V H The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:4-17 H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain only one or more conserved substitutions in HCDR1, HCDR2, and HCDR3 (e.g., only one or more highly conserved substitutions); or

[0497] b)V L The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-30 L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions), or

[0498] Both a) and b)

[0499] In some embodiments, the polypeptide comprises:

[0500] a)V H The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:4-17 H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain only one or more conserved substitutions in HCDR1, HCDR2, and HCDR3 (e.g., only one or more highly conserved substitutions); and

[0501] b)V L The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-30 LThe amino acid sequences of LCDR1, LCDR2, and LCDR3 contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions).

[0502] In some embodiments, the polypeptide comprises:

[0503] a)V H The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:4-17 H The amino acid sequences HCDR1, HCDR2, and HCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions); or

[0504] b)V L The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-30 L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions), or

[0505] Both a) and b)

[0506] In some embodiments, the polypeptide comprises:

[0507] a)V H The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:4-17 H The amino acid sequences HCDR1, HCDR2, and HCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions); and

[0508] b)V L The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-30 L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions).

[0509] In some embodiments, the polypeptide comprises:

[0510] a)V H The amino acid sequence contains V that is respectively associated with one or more of the sequences in SEQ ID NO:4-17. HThe amino acid sequences of HCDR1, HCDR2, and HCDR3 have 100% sequence identity; or

[0511] b)V L The amino acid sequence contains V that is respectively associated with one or more of SEQ ID NO:20-30. L LCDR1, LCDR2, and LCDR3 have 100% sequence identity, or

[0512] Both a) and b)

[0513] In some embodiments, the polypeptide comprises:

[0514] a)V H The amino acid sequence contains V that is respectively associated with any one or more of SEQ ID NO:4-17. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 have 100% sequence identity; and

[0515] b)V L The amino acid sequence contains V that is respectively associated with one or more of SEQ ID NO:20-30. L The amino acid sequences of LCDR1, LCDR2, and LCDR3 have 100% sequence identity.

[0516] In some embodiments, the peptide comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of an antibody, wherein the antibody comprises V selected from the group consisting of V. H / V L combination:

[0517] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0518] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0519] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0520] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0521] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0522] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0523] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0524] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0525] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0526] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0527] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0528] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0529] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0530] SEQ ID NO:17 and SEQ ID NO:30(AB-14).

[0531] In some embodiments, the polypeptide comprises:

[0532] a) HCDR1, which contains the amino acid sequence shown in SEQ ID NO:31;

[0533] b) HCDR2, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:33-46 (e.g., at least one amino acid sequence shown in SEQ ID NO:34-46);

[0534] c) HCDR3, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:48-57 (e.g., at least one amino acid sequence shown in SEQ ID NO:49-57);

[0535] d) LCDR1, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:59-66 (e.g., at least one amino acid sequence shown in SEQ ID NO:60-66);

[0536] e) LCDR2, which contains the amino acid sequence shown in SEQ ID NO:67;

[0537] f) LCDR3, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:69-80 (e.g., at least one amino acid sequence shown in SEQ ID NO:70-80);

[0538] Or any combination thereof.

[0539] In some embodiments, the polypeptide comprises:

[0540] a)V H The amino acid sequence contains V that corresponds to any one of the amino acids shown in SEQ ID NO:4-7. H HCDR1, HCDR2 and HCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity);

[0541] b)V L The amino acid sequence contains V that corresponds to any one of the amino acids shown in SEQ ID NO:20-22. L LCDR1, LCDR2, and LCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity), or

[0542] Both a) and b)

[0543] In some embodiments, the polypeptide comprises:

[0544] a)V H The amino acid sequence contains V that is respectively associated with any one or more of SEQ ID NO:4-7. H HCDR1, HCDR2 and HCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity);

[0545] b)V L The amino acid sequence contains V that is respectively associated with one or more of the sequences in SEQ ID NO:20-22. L LCDR1, LCDR2, and LCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity), or

[0546] Both a) and b)

[0547] In some embodiments, the polypeptide comprises:

[0548] a)V HThe amino acid sequence contains V that is respectively associated with any one or more of SEQ ID NO:4-7. H HCDR1, HCDR2, and HCDR3 with fundamentally similar amino acid sequences (e.g., having at least about 90% sequence identity); and

[0549] b)V L The amino acid sequence contains V that is respectively associated with one or more of the sequences in SEQ ID NO:20-22. L LCDR1, LCDR2, and LCDR3 have fundamentally similar amino acid sequences (e.g., having at least about 90% sequence identity).

[0550] In some embodiments, the polypeptide comprises:

[0551] a)V H The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:4-7 respectively. H The amino acid sequence of HCDR1, HCDR2, and HCDR3 has one or more functional characteristics of HCDR1, HCDR2, and HCDR3; or

[0552] b)V L The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:20-22 respectively. L The amino acid sequence of LCDR1, LCDR2, and LCDR3 contains one or more functional characteristics of LCDR1, LCDR2, and LCDR3, or

[0553] Both a) and b)

[0554] In some embodiments, the polypeptide comprises:

[0555] a)V H The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:4-7 respectively. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain one or more functional characteristics of HCDR1, HCDR2, and HCDR3; and

[0556] b)V L The amino acid sequence, which contains substantially one or more of the V values ​​from SEQ ID NO:20-22 respectively. L One or more functional properties of the amino acid sequences of LCDR1, LCDR2 and LCDR3.

[0557] In some embodiments, the polypeptide comprises:

[0558] a)V H The amino acid sequence contains V corresponding to any one or more of SEQ ID NO:4-7. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain only one or more conserved substitutions in HCDR1, HCDR2, and HCDR3 (e.g., only one or more highly conserved substitutions); or

[0559] b)V L The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-22 L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions), or

[0560] Both a) and b)

[0561] In some embodiments, the polypeptide comprises:

[0562] a)V H The amino acid sequence contains V corresponding to any one or more of SEQ ID NO:4-7. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain only one or more conserved substitutions in HCDR1, HCDR2, and HCDR3 (e.g., only one or more highly conserved substitutions); and

[0563] b)V L The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-22 L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions).

[0564] In some embodiments, the polypeptide comprises:

[0565] a)V H The amino acid sequence contains V corresponding to any one or more of SEQ ID NO:4-7. H The amino acid sequences HCDR1, HCDR2, and HCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions); or

[0566] b)V LThe amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-22 L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions), or

[0567] Both a) and b)

[0568] In some embodiments, the polypeptide comprises:

[0569] a)V H The amino acid sequence contains V corresponding to any one or more of SEQ ID NO:4-7. H The amino acid sequences HCDR1, HCDR2, and HCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions); and

[0570] b)V L The amino acid sequence comprising V, respectively, relative to any one or more of SEQ ID NO:20-22 L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions).

[0571] In some embodiments, the polypeptide comprises:

[0572] a)V H The amino acid sequence contains V that is respectively associated with any one or more of SEQ ID NO:4-7. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 have 100% sequence identity; or

[0573] b)V L The amino acid sequence contains V that is respectively associated with one or more of the sequences in SEQ ID NO:20-22. L LCDR1, LCDR2, and LCDR3 have 100% sequence identity, or

[0574] Both a) and b)

[0575] In some embodiments, the polypeptide comprises:

[0576] a)V HThe amino acid sequence contains V that is respectively associated with any one or more of SEQ ID NO:4-7. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 have 100% sequence identity; and

[0577] b)V L The amino acid sequence contains V that is respectively associated with one or more of the sequences in SEQ ID NO:20-22. L The amino acid sequences of LCDR1, LCDR2, and LCDR3 have 100% sequence identity.

[0578] In some embodiments, the peptide comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of an antibody, wherein the antibody comprises V selected from the group consisting of V. H / V L combination:

[0579] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0580] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0581] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0582] SEQ ID NO:7 and SEQ ID NO:22(AB-4).

[0583] In some embodiments, the polypeptide comprises:

[0584] a) HCDR1, which contains the amino acid sequence shown in SEQ ID NO:31;

[0585] b) HCDR2, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:33-37 (e.g., at least one amino acid sequence shown in SEQ ID NO:34-37);

[0586] c) HCDR3, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:48-52 (e.g., at least one amino acid sequence shown in SEQ ID NO:49-52);

[0587] d) LCDR1, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:59-61 (e.g., at least one amino acid sequence shown in SEQ ID NO:60 or SEQ ID NO:61);

[0588] e) LCDR2, which contains the amino acid sequence shown in SEQ ID NO:67;

[0589] f) LCDR3, which contains at least one amino acid substitution relative to at least one amino acid sequence shown in SEQ ID NO:69-72 (e.g., at least one amino acid sequence shown in SEQ ID NO:70-72);

[0590] Or any combination thereof.

[0591] In some embodiments, the polypeptide comprises:

[0592] a)V H The amino acid sequence contains V, which is respectively associated with SEQ ID NO:5. H HCDR1, HCDR2 and HCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity);

[0593] b)V L The amino acid sequence contains V, which is respectively associated with SEQ ID NO:21. L LCDR1, LCDR2, and LCDR3 have amino acid sequences that are substantially similar to each other (e.g., have at least about 90% sequence identity), or

[0594] Both a) and b)

[0595] In some embodiments, the polypeptide comprises:

[0596] a)V H The amino acid sequence contains V, which is respectively associated with SEQ ID NO:5. H HCDR1, HCDR2, and HCDR3 with fundamentally similar amino acid sequences (e.g., having at least about 90% sequence identity); and

[0597] b)V L The amino acid sequence contains V, which is respectively associated with SEQ ID NO:21. L LCDR1, LCDR2, and LCDR3 have fundamentally similar amino acid sequences (e.g., having at least about 90% sequence identity).

[0598] In some embodiments, the polypeptide comprises:

[0599] a)V H The amino acid sequence, which contains essentially the V of SEQ ID NO:5, respectively. H The amino acid sequence of HCDR1, HCDR2, and HCDR3 has one or more functional characteristics of HCDR1, HCDR2, and HCDR3; or

[0600] b)V L The amino acid sequence, which contains essentially the V of SEQ ID NO:21, respectively. L The amino acid sequence of LCDR1, LCDR2, and LCDR3 contains one or more functional characteristics of LCDR1, LCDR2, and LCDR3, or

[0601] Both a) and b)

[0602] In some embodiments, the polypeptide comprises:

[0603] a)V H The amino acid sequence, which contains essentially the V of SEQ ID NO:5, respectively. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain one or more functional characteristics of HCDR1, HCDR2, and HCDR3; and

[0604] b)V L The amino acid sequence, which contains essentially the V of SEQ ID NO:21, respectively. L One or more functional properties of the amino acid sequences of LCDR1, LCDR2 and LCDR3.

[0605] In some embodiments, the polypeptide comprises:

[0606] a)V H The amino acid sequence, which contains V corresponding to SEQ ID NO:5 respectively H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain only one or more conserved substitutions in HCDR1, HCDR2, and HCDR3 (e.g., only one or more highly conserved substitutions); or

[0607] b)V L The amino acid sequence, which contains V corresponding to SEQ ID NO:21 respectively LThe amino acid sequences of LCDR1, LCDR2, and LCDR3 contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions), or

[0608] Both a) and b)

[0609] In some embodiments, the polypeptide comprises:

[0610] a)V H The amino acid sequence, which contains V corresponding to SEQ ID NO:5 respectively H The amino acid sequences of HCDR1, HCDR2, and HCDR3 contain only one or more conserved substitutions in HCDR1, HCDR2, and HCDR3 (e.g., only one or more highly conserved substitutions); and

[0611] b)V L The amino acid sequence, which contains V corresponding to SEQ ID NO:21 respectively L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions).

[0612] In some embodiments, the polypeptide comprises:

[0613] a)V H The amino acid sequence, which contains V corresponding to SEQ ID NO:5 respectively H The amino acid sequences HCDR1, HCDR2, and HCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions); or

[0614] b)V L The amino acid sequence, which contains V corresponding to SEQ ID NO:21 respectively L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions), or

[0615] Both a) and b)

[0616] In some embodiments, the polypeptide comprises:

[0617] a)V H The amino acid sequence, which contains V corresponding to SEQ ID NO:5 respectively HThe amino acid sequences HCDR1, HCDR2, and HCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions); and

[0618] b)V L The amino acid sequence, which contains V corresponding to SEQ ID NO:21 respectively L The amino acid sequences of LCDR1, LCDR2, and LCDR3 contain up to 1, 2, or 3 conserved substitutions (e.g., up to 1, 2, or 3 highly conserved substitutions).

[0619] In some embodiments, the polypeptide comprises:

[0620] a)V H The amino acid sequence contains V, which is respectively associated with SEQ ID NO:5. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 have 100% sequence identity; or

[0621] b)V L The amino acid sequence contains V, which is respectively associated with SEQ ID NO:21. L LCDR1, LCDR2, and LCDR3 have 100% sequence identity, or

[0622] Both a) and b)

[0623] In some embodiments, the polypeptide comprises:

[0624] a)V H The amino acid sequence contains V, which is respectively associated with SEQ ID NO:5. H The amino acid sequences of HCDR1, HCDR2, and HCDR3 have 100% sequence identity; and

[0625] b)V L The amino acid sequence contains V, which is respectively associated with SEQ ID NO:21. L The amino acid sequences of LCDR1, LCDR2, and LCDR3 have 100% sequence identity.

[0626] In some embodiments, the polypeptide comprises:

[0627] a)V HIt contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:4, and

[0628] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:20(AB-1).

[0629] In some embodiments, the polypeptide comprises:

[0630] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:5; and

[0631] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:21(AB-2).

[0632] In some embodiments, the polypeptide comprises:

[0633] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:6; and

[0634] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:20(AB-3).

[0635] In some embodiments, the polypeptide comprises:

[0636] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:7; and

[0637] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:22(AB-4).

[0638] In some embodiments, the polypeptide comprises:

[0639] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:8; and

[0640] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:23(AB-5).

[0641] In some embodiments, the polypeptide comprises:

[0642] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:9; and

[0643] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:24(AB-6).

[0644] In some embodiments, the polypeptide comprises:

[0645] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:10; and

[0646] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:25(AB-7).

[0647] In some embodiments, the polypeptide comprises:

[0648] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:11; and

[0649] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:20(AB-8).

[0650] In some embodiments, the polypeptide comprises:

[0651] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:12; and

[0652] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:20(AB-9).

[0653] In some embodiments, the polypeptide comprises:

[0654] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:13; and

[0655] b)V LIt contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:26(AB-10).

[0656] In some embodiments, the polypeptide comprises:

[0657] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:14; and

[0658] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:27(AB-11).

[0659] In some embodiments, the polypeptide comprises:

[0660] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:15; and

[0661] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:28(AB-12).

[0662] In some embodiments, the polypeptide comprises:

[0663] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:16; and

[0664] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:29(AB-13).

[0665] In some embodiments, the polypeptide comprises:

[0666] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:17; and

[0667] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:30(AB-14).

[0668] In some embodiments, the polypeptide comprises:

[0669] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0670] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:34;

[0671] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:49;

[0672] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:60;

[0673] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0674] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:70, or any combination thereof.

[0675] In some embodiments, the polypeptide comprises:

[0676] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0677] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:35;

[0678] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:50;

[0679] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:60;

[0680] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0681] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:71, or any combination thereof.

[0682] In some embodiments, the polypeptide comprises:

[0683] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0684] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:36;

[0685] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:51;

[0686] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:60;

[0687] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0688] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:70, or any combination thereof.

[0689] In some embodiments, the polypeptide comprises:

[0690] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0691] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:37;

[0692] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:52;

[0693] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:61;

[0694] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0695] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:72, or any combination thereof.

[0696] In some embodiments, the polypeptide comprises:

[0697] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0698] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:38;

[0699] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:53;

[0700] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:59;

[0701] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0702] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:73, or any combination thereof.

[0703] In some embodiments, the polypeptide comprises:

[0704] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0705] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:39;

[0706] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:53;

[0707] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:60;

[0708] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0709] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:74, or any combination thereof.

[0710] In some embodiments, the polypeptide comprises:

[0711] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0712] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:40;

[0713] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:49;

[0714] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:62;

[0715] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0716] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:75, or any combination thereof.

[0717] In some embodiments, the polypeptide comprises:

[0718] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0719] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:41;

[0720] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:54;

[0721] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:60;

[0722] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0723] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:70, or any combination thereof.

[0724] In some embodiments, the polypeptide comprises:

[0725] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0726] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:41;

[0727] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:51;

[0728] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:60;

[0729] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0730] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:70, or any combination thereof.

[0731] In some embodiments, the polypeptide comprises:

[0732] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0733] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:42;

[0734] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:53;

[0735] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:63;

[0736] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0737] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:76, or any combination thereof.

[0738] In some embodiments, the polypeptide comprises:

[0739] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0740] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:43;

[0741] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:55;

[0742] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:64;

[0743] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0744] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:77, or any combination thereof.

[0745] In some embodiments, the polypeptide comprises:

[0746] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0747] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:44;

[0748] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:56;

[0749] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:65;

[0750] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0751] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:78, or

[0752] Any of the aforementioned combinations.

[0753] In some embodiments, the polypeptide comprises:

[0754] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0755] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:45;

[0756] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:57;

[0757] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:66;

[0758] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0759] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:79, or

[0760] Any of the aforementioned combinations.

[0761] In some embodiments, the polypeptide comprises:

[0762] a) HCDR1, which contains the amino acid sequence of SEQ ID NO:31;

[0763] b) HCDR2, which contains the amino acid sequence of SEQ ID NO:46;

[0764] c) HCDR3, which contains the amino acid sequence of SEQ ID NO:48;

[0765] d) LCDR1, which contains the amino acid sequence of SEQ ID NO:63;

[0766] e) LCDR2, which contains the amino acid sequence of SEQ ID NO:67;

[0767] f) LCDR3, which contains the amino acid sequence of SEQ ID NO:80, or

[0768] Any of the aforementioned combinations.

[0769] complementary position

[0770] The amino acid residues at the complementary site facilitate the interaction between the antibody and its target protein epitope. The interaction can be a hydrogen bond, salt bridge, van der Waals interaction, electrostatic interaction, hydrophobic interaction, π-interaction effect, ionic bond, and / or any combination thereof. The interaction can be direct or indirect, for example, through a coordinating intermediate molecule, such as an ion or water. In some embodiments, the residues at the complementary site comprise only residues that are part of a defined CDR. In some embodiments, the residues at the complementary site further comprise one or more residues that are not part of a defined CDR (e.g., residues defining a frame region).

[0771] In some embodiments, when the polypeptide binds to the target antigen, the distance between the complementary site and the epitope on the target antigen is less than about 5.0 Å, for example 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 Å, or about 0.9-5.0 or 0.9-4.8 Å from the epitope. 1.0-5, 1.0-4.5, 1.0-4.0, 1.0-3.5, 1.1-3.5, 1.1-3.0, 1.2-3.0, 1.2-2.5, 1.3-2.5, 1.3-2.4, 1.4-2.4, 1.4-2.3, 1.5-2.3, 1.5-2.2, 1.6-2.2, 1.6-2.1, 1.7-2.1, 1.7-2.0 or 1.8-2.0 angstroms. In some embodiments, when the polypeptide binds to the target antigen, the distance between less than all of the amino acid residues constituting the complementary site (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) and the epitope on the target antigen is less than about 5.0 Å.

[0772] In some embodiments, a polypeptide comprising the complementary site disclosed herein (e.g., an antibody or its antigen-binding fragment) comprises V H and V L In some embodiments, complementary residues are contained in the V of the polypeptide. H and V L Inside.

[0773] In some embodiments, the polypeptide comprises V containing SEQ ID NO:3 / SEQ ID NO:19 H / V L The complementary sites of the combined antibodies are different complementary sites.

[0774] In some embodiments, the polypeptide comprises, via substitution of 1 to 3 residues (e.g., 1, 2, or 3), a conserved substitution such as a highly conserved substitution, with respect to V containing SEQ ID NO:3 / SEQ ID NO:19. H / V L The complementary sites of the combined antibodies are different complementary sites.

[0775] In some embodiments, the polypeptide comprises V containing SEQ ID NO:3 / SEQ ID NO:19 H / V L The complementary sites of the combined antibodies are substantially similar (e.g., having at least about 90% sequence identity). In some embodiments, the polypeptide comprises V containing SEQ ID NO:3 / SEQ ID NO:19.H / V L The complementary sites of the combined antibodies are substantially similar (e.g., have at least about 90% sequence identity) and substantially retain one or more of their functional properties.

[0776] In some embodiments, the polypeptide comprises V containing SEQ ID NO:3 / SEQ ID NO:19 H / V L The complementary sites of the combined antibodies have 100% sequence identity.

[0777] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V L The complementary sites of the combined antibodies are different: 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).

[0778] See Table 1 for SEQ ID NO:4-17, and V containing any one of SEQ ID NO:4-17. H The complementary residues of the antibody sequence are shown in [link to antibody sequence]. Figure 2A-2B See Table 2 for SEQ ID NO:20-30.

[0779] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V LThe complementary sites of the combined antibodies are different: 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), by substitution of 1 to 3 (e.g., 1, 2 or 3) residues (e.g., conservative substitution, such as highly conservative substitution).

[0780] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V L The complementary sites of the combined antibodies are substantially similar (e.g., having at least about 90% sequence identity):

[0781] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0782] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0783] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0784] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0785] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0786] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0787] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0788] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0789] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0790] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0791] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0792] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0793] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0794] SEQ ID NO:17 and SEQ ID NO:30 (AB-14), or

[0795] Its combination.

[0796] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V L The complementary sites of the combined antibodies are substantially similar (e.g., have at least about 90% sequence identity) and substantially retain one or more of their functional properties:

[0797] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0798] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0799] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0800] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0801] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0802] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0803] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0804] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0805] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0806] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0807] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0808] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0809] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0810] SEQ ID NO:17 and SEQ ID NO:30 (AB-14), or

[0811] Its combination.

[0812] In some embodiments, relative to V which includes the following selections H / V L The complementary sites of the combined antibodies, the peptides contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions):

[0813] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0814] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0815] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0816] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0817] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0818] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0819] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0820] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0821] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0822] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0823] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0824] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0825] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0826] SEQ ID NO:17 and SEQ ID NO:30 (AB-14), or

[0827] Its combination.

[0828] In some embodiments, relative to V which includes the following selections H / V L The complementary sites of the combined antibodies, the peptides containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 conserved substitutions (e.g., up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 highly conserved substitutions):

[0829] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0830] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0831] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0832] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0833] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0834] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0835] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0836] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0837] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0838] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0839] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0840] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0841] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0842] SEQ ID NO:17 and SEQ ID NO:30 (AB-14), or

[0843] Its combination.

[0844] In some embodiments, the polypeptide comprises V selected from the following: H / V L The complementary sites of the combined antibodies have 100% sequence identity:

[0845] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0846] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0847] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[0848] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[0849] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[0850] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[0851] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[0852] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[0853] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[0854] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[0855] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[0856] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[0857] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[0858] SEQ ID NO:17 and SEQ ID NO:30 (AB-14), or

[0859] Its combination.

[0860] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V L The complementary sites of the combined antibodies are different: 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).

[0861] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V L The complementary sites of the combined antibodies are different: 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), by substitution of 1 to 3 (e.g. 1, 2 or 3) residues (e.g., conservative substitution, such as highly conserved substitution).

[0862] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V L The complementary sites of the combined antibodies are substantially similar (e.g., having at least about 90% sequence identity):

[0863] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0864] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0865] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0866] SEQ ID NO:7 and SEQ ID NO:22(AB-4), or

[0867] Its combination.

[0868] In some embodiments, the polypeptide comprises V containing a subset of the following. H / V L The complementary sites of the combined antibodies are substantially similar (e.g., have at least about 90% sequence identity) and substantially retain one or more of their functional properties:

[0869] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0870] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0871] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0872] SEQ ID NO:7 and SEQ ID NO:22(AB-4), or

[0873] Its combination.

[0874] In some embodiments, relative to V which includes the following selections H / V L The complementary sites of the combined antibodies, the peptides contain only one or more conserved substitutions (e.g., only one or more highly conserved substitutions):

[0875] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0876] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0877] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0878] SEQ ID NO:7 and SEQ ID NO:22(AB-4), or

[0879] Its combination.

[0880] In some embodiments, relative to V which includes the following selections H / V L The complementary sites of the combined antibodies, the peptides containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 conserved substitutions (e.g., up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 highly conserved substitutions):

[0881] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0882] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0883] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0884] SEQ ID NO:7 and SEQ ID NO:22(AB-4), or

[0885] Its combination.

[0886] In some embodiments, the polypeptide comprises V selected from the following H / V L The complementary sites of the combined antibodies have 100% sequence identity:

[0887] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[0888] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[0889] SEQ ID NO:6 and SEQ ID NO:20(AB-3); or

[0890] SEQ ID NO:7 and SEQ ID NO:22(AB-4), or

[0891] Its combination.

[0892] In some embodiments, the polypeptide includes a complementary site comprising an amino acid residue, or a subset thereof, corresponding to each of M2, G26, F27, T28, R30, T31, Y32, Y53, R98, P100, Q101, W102, E103, and E107 of SEQ ID NO:2. In some embodiments, the polypeptide includes a complementary site comprising an amino acid residue, or a subset thereof, corresponding to each of T28, T31, Y32, Y53, R98, W102, and E107 of SEQ ID NO:2.

[0893] consensus sequence

[0894] In some embodiments, the polypeptide (e.g., an antibody or its antigen-binding fragment) comprises V containing the amino acid sequence of SEQ ID NO:2. H ,in:

[0895] X1 is not I;

[0896] X2 is not W;

[0897] X3 is not D;

[0898] X4 is not S;

[0899] X5 is not N;

[0900] X6 is not K;

[0901] X7 is not A;

[0902] X8 is not A;

[0903] X9 is not L;

[0904] X 10 Not V;

[0905] X 11 Not H;

[0906] X 12 Not A;

[0907] X 13 Not F; or

[0908] X 14 Not I, or

[0909] Any of the aforementioned combinations.

[0910] The sequence identified as SEQ ID NO:2 is shown in Table 1, and it is a common sequence of SEQ ID NO:3-17 in this paper. H sequence.

[0911] In some embodiments:

[0912] X1 is either I or V;

[0913] X2 is either W or S;

[0914] X3 is either D or S;

[0915] X4 is S, T, Y, or A;

[0916] X5 is N, Y, D, or T;

[0917] X6 is K, T, or I;

[0918] X7 is A, S, or T;

[0919] X8 is either A or S;

[0920] X9 is L, D, E, or S;

[0921] X 10 It is V or I;

[0922] X 11 It is H, F, or Y;

[0923] X 12 It is either A or S;

[0924] X 13 It is F, M, or L;

[0925] X 14 Is it I or V?

[0926] Or any combination thereof.

[0927] In some embodiments:

[0928] X1 is V;

[0929] X2 is S;

[0930] X3 is S;

[0931] X4 is T, Y, or A;

[0932] X5 is Y, D, or T;

[0933] X6 is either T or I;

[0934] X7 is either S or T;

[0935] X8 is S;

[0936] X9 is D, E, or S;

[0937] X 10 It's I;

[0938] X 11 It is either F or Y;

[0939] X 12 It is S;

[0940] X 13 Is it M or L;

[0941] X 14 It's V.

[0942] Or any combination thereof.

[0943] 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.

[0944] 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.

[0945] 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.

[0946] 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.

[0947] 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.

[0948] 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.

[0949] 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.

[0950] 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.

[0951] 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.

[0952] In some embodiments, X 10 Not V. In some embodiments, X 10 It is V or I. In some embodiments, X 10 It is I. In some embodiments, X 10 It is V.

[0953] In some embodiments, X 11 Not H. In some embodiments, X 11 It is H, F, or Y. In some embodiments, X 11 It is F or Y. In some embodiments, X 11 It is F. In some embodiments, X 11 It is Y. In some embodiments, X 11 It’s H.

[0954] In some embodiments, X 12 Not A. In some embodiments, X 12 It is A or S. In some embodiments, X 12 It is S. In some embodiments, X 12 It’s A.

[0955] In some embodiments, X 13 Not F. In some embodiments, X 13 It is F, M, or L. In some embodiments, X 13 It is M or L. In some embodiments, X 13 It is M. In some embodiments, X 13 It is L. In some embodiments, X 13 It's F.

[0956] In some embodiments, X 14 Not I. In some embodiments, X 14 It is I or V. In some embodiments, X 14 It is V. In some embodiments, X 14 It's I.

[0957] In some embodiments, the polypeptide comprises V containing the amino acid sequence of SEQ ID NO:18. L ,in:

[0958] X 15 Not N;

[0959] X 16 Not L;

[0960] X 17 Not S;

[0961] X 18 Not K;

[0962] X 19 Not S;

[0963] X 20 Not V;

[0964] X 21 Not W;

[0965] X 22 Not D;

[0966] X 23 Not S;

[0967] X 24 Not S;

[0968] X 25 Not S;

[0969] X 26 Not D; or

[0970] X 27 Not H, or

[0971] Any of the aforementioned combinations.

[0972] The sequence identified as SEQ ID NO:18 is shown in Table 2, and it is a common sequence of SEQ ID NO:19-30 in this paper. L sequence.

[0973] In some embodiments:

[0974] X 15 It is either N or Y;

[0975] X 16 It is L or I;

[0976] X 17 It is S or R;

[0977] X 18 Is it K, F, or Y?

[0978] X 19 It is S or N;

[0979] X 20 It is V or I;

[0980] X 21 It is W or Y;

[0981] X 22 Is it D, V, or S?

[0982] X 23 Is it S, M, or E?

[0983] X 24 It is S, A, or T;

[0984] X 25 It is S, D, or E;

[0985] X 26 It is D, S, R, or F;

[0986] X 27 Is it H, L, K, or E?

[0987] Or any combination thereof.

[0988] In some embodiments:

[0989] X 15 It is Y;

[0990] X 16 It's I;

[0991] X 17 It is R;

[0992] X 18 It is either F or Y;

[0993] X 19 It is N;

[0994] X 20 It's I;

[0995] X 21 It is Y;

[0996] X 22 Is it V or S;

[0997] X 23 It is M or E;

[0998] X 24 It is either A or T;

[0999] X 25 It is either D or E;

[1000] X 26 Is it S, R, or F?

[1001] X 27 Is it L, K, or E?

[1002] Or any combination thereof.

[1003] In some embodiments, X 15Not N. In some embodiments, X 15 It is N or Y. In some embodiments, X 15 It is Y. In some embodiments, X 15 It's N.

[1004] In some embodiments, X 16 Not L. In some embodiments, X 16 It is L or I. In some embodiments, X 16 It is I. In some embodiments, X 16 It is L.

[1005] In some embodiments, X 17 Not S. In some embodiments, X 17 It is S or R. In some embodiments, X 17 It is R. In some embodiments, X 17 It’s S.

[1006] In some embodiments, X 18 Not K. In some embodiments, X 18 It is K, F, or Y. In some embodiments, X 18 It is F or Y. In some embodiments, X 18 It is F. In some embodiments, X 18 It is Y. In some embodiments, X 18 It's K.

[1007] In some embodiments, X 19 Not S. In some embodiments, X 19 It is S or N. In some embodiments, X 19 It is N. In some embodiments, X 19 It’s S.

[1008] In some embodiments, X 20 Not V. In some embodiments, X 20 It is V or I. In some embodiments, X 20 It is I. In some embodiments, X 20 It is V.

[1009] In some embodiments, X 21 Not W. In some embodiments, X 21 It is W or Y. In some embodiments, X 21 It is Y. In some embodiments, X 21 It is W.

[1010] In some embodiments, X 22 Not D. In some embodiments, X 22 It is D, V, or S. In some embodiments, X22 It is V or S. In some embodiments, X 22 It is V. In some embodiments, X 22 It is S. In some embodiments, X 22 It is D.

[1011] In some embodiments, X 23 Not S. In some embodiments, X 23 It is S, M, or E. In some embodiments, X 23 It is M or E. In some embodiments, X 23 It is M. In some embodiments, X 23 It is E. In some embodiments, X 23 It’s S.

[1012] In some embodiments, X 24 Not S. In some embodiments, X 24 It is S, A, or T. In some embodiments, X 24 It is A or T. In some embodiments, X 24 It is A. In some embodiments, X 24 It is T. In some embodiments, X 24 It’s S.

[1013] In some embodiments, X 25 Not S. In some embodiments, X 25 It is S, D, or E. In some embodiments, X 25 It is D or E. In some embodiments, X 25 It is D. In some embodiments, X 25 It is E. In some embodiments, X 25 It’s S.

[1014] In some embodiments, X 26 Not D. In some embodiments, X 26 It is D, S, R, or F. In some embodiments, X 26 It is S, R, or F. In some embodiments, X 26 It is S. In some embodiments, X 26 It is R. In some embodiments, X 26 It is F. In some embodiments, X 26 It is D.

[1015] In some embodiments, X 27 Not H. In some embodiments, X 27 It is H, L, K, or E. In some embodiments, X 27 It is L, K, or E. In some embodiments, X 27 It is L. In some embodiments, X27 It is K. In some embodiments, X 27 It is E. In some embodiments, X 27 It’s H.

[1016] In some embodiments,

[1017] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is M, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1018] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is M, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is I, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1019] 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, X 10 It is I, X 11 It is F, X 12 It is A, X13 It is F, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1020] 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, X 10 It is I, X 11 It is F, X 12 It is S, X 13 It is M, X 14 It is V, X 15 It is Y, X 16 It is I, X 17 It is S, X 18 It is F, X 19 It is S, X 20 It is I, X 21 It is Y, X 22 It is V, X 23 It is S, X 24 It is A, X 25 It is S, X 26 Is it R or X? 27 It is L, or a combination thereof;

[1021] 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, X 10 It is I, X 11 It is F, X 12 It is S, X 13 It is M, X 14 It is I, X 15 It is N, X 16 It is L, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is M, X 24 It is S, X 25 It is S, X 26Is it D or X? 27 It is L, or a combination thereof;

[1022] 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, X 10 It is I, X 11 It is F, X 12 It is S, X 13 It is M, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is I, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it D or X? 27 It is H, or a combination thereof;

[1023] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is M, X 14 It is I, X 15 It is N, X 16 It is L, X 17 It is S, X 18 It is Y, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is M, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1024] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is F, X 14 It is I, X 15 It is N, X 16 It is I, X 17It is S, X 18 It is K, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1025] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is F, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1026] 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, X 10 It is I, X 11 It is F, X 12 It is S, X 13 It is M, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is N, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is D, X 26 Is it F or X? 27 It is L, or a combination thereof;

[1027] 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, X 10 It is V, X 11 It is Y, X 12 It is A, X 13 It is L, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is F, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is S, X 23 It is S, X 24 It is T, X 25 It is S, X 26 Is it R or X? 27 It is H, or a combination thereof;

[1028] 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, X 10 It is V, X 11 It is H, X 12 It is A, X 13 It is F, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is R, X 18 It is F, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is V, X 23 It is S, X 24 It is T, X 25 It is S, X 26 Is it D or X? 27 It is K, or a combination thereof;

[1029] 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, X 10 It is V, X 11 It is Y, X 12 It is A, X 13 It is F, X 14 It is I, X 15 It is N, X 16 It is L, X 17 It is S, X 18 It is F, X 19 It is S, X 20It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is E, X 26 Is it D or X? 27 It is L, or a combination thereof; or

[1030] (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, X 10 It is V, X 11 It is H, X 12 It is A, X 13 It is F, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is N, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is E, X 24 It is S, X 25 It is S, X 26 Is it D or X? 27 It is E, or a combination thereof.

[1031] In some embodiments:

[1032] X1 is either I or V;

[1033] X2 is W;

[1034] X3 is either D or S;

[1035] X4 is either S or T;

[1036] X5 is N, Y, or D;

[1037] X6 is either K or T;

[1038] X7 is either A or S;

[1039] X8 is either A or S;

[1040] X9 is L, D, E, or S;

[1041] X 10 It is V or I;

[1042] X 11 It is H or F;

[1043] X 12 It is either A or S;

[1044] X 13 Is it F or M; or

[1045] X 14 Is it I or V, or

[1046] Any of the aforementioned combinations.

[1047] In some embodiments:

[1048] X1 is V;

[1049] X2 is W;

[1050] X3 is S;

[1051] X4 is T;

[1052] X5 is either Y or D;

[1053] X6 is T;

[1054] X7 is S;

[1055] X8 is S;

[1056] X9 is D, E, or S;

[1057] X 10 It's I;

[1058] X 11 It is F;

[1059] X 12 It is S;

[1060] X 13 It is M;

[1061] X 14 It's V.

[1062] Or any combination thereof.

[1063] 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.

[1064] In some embodiments, X2 is W.

[1065] 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.

[1066] 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.

[1067] 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.

[1068] 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.

[1069] 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.

[1070] 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.

[1071] 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.

[1072] In some embodiments, X 10 Not V. In some embodiments, X 10 It is V or I. In some embodiments, X 10 It is I. In some embodiments, X 10 It is V.

[1073] In some embodiments, X 11 Not H. In some embodiments, X 11 It is H or F. In some embodiments, X 11 It is F. In some embodiments, X 11 It’s H.

[1074] In some embodiments, X 12 Not A. In some embodiments, X 12 It is A or S. In some embodiments, X 12 It is S. In some embodiments, X 12 It’s A.

[1075] In some embodiments, X 13 Not F. In some embodiments, X 13 It is F or M. In some embodiments, X 13 It is M. In some embodiments, X 13 It's F.

[1076] In some embodiments, X 14 Not I. In some embodiments, X 14 It is I or V. In some embodiments, X 14 It is V. In some embodiments, X 14 It's I.

[1077] In some embodiments:

[1078] X 15 It is either N or Y;

[1079] X 16 It is L or I;

[1080] X 17 It is S;

[1081] X 18 It is K or F;

[1082] X 19 It is S;

[1083] X 20 It is V or I;

[1084] X 21 It is W or Y;

[1085] X 22 It is D or V;

[1086] X 23 It is S;

[1087] X 24 It is S or A;

[1088] X 25 It is S;

[1089] X 26 Is it D, S, or R?

[1090] X 27 Is it H or L?

[1091] Or any combination thereof.

[1092] In some embodiments:

[1093] X 15 It is Y;

[1094] X 16 It's I;

[1095] X 17 It is S;

[1096] X 18 It is F;

[1097] X 19 It is S;

[1098] X 20 It's I;

[1099] X 21 It is Y;

[1100] X 22 It is V;

[1101] X 23 It is S;

[1102] X 24 It is A;

[1103] X 25 It is S;

[1104] X 26 It is S or R;

[1105] X 27 It is L.

[1106] Or any combination thereof.

[1107] In some embodiments, X 15 Not N. In some embodiments, X 15 It is N or Y. In some embodiments, X 15 It is Y. In some embodiments, X 15 It's N.

[1108] In some embodiments, X 16 Not L. In some embodiments, X 16 It is L or I. In some embodiments, X 16 It is I. In some embodiments, X 16 It is L.

[1109] In some embodiments, X 17 It’s S.

[1110] In some embodiments, X 18 Not K. In some embodiments, X 18 It is K or F. In some embodiments, X 18 It is F. In some embodiments, X 18 It's K.

[1111] In some embodiments, X 19 It’s S.

[1112] In some embodiments, X 20 Not V. In some embodiments, X 20 It is V or I. In some embodiments, X 20 It is I. In some embodiments, X 20 It is V.

[1113] In some embodiments, X 21 Not W. In some embodiments, X 21 It is W or Y. In some embodiments, X 21 It is Y. In some embodiments, X 21 It is W.

[1114] In some embodiments, X 22 Not D. In some embodiments, X 22 It is D or V. In some embodiments, X 22 It is V. In some embodiments, X 22 It is D.

[1115] In some embodiments, X 23 It’s S.

[1116] In some embodiments, X 24 Not S. In some embodiments, X 24 It is S or A. In some embodiments, X 24 It is A. In some embodiments, X 24 It’s S.

[1117] In some embodiments, X 25 It’s S.

[1118] In some embodiments, X 26 Not D. In some embodiments, X 26 It is D, S, or R. In some embodiments, X 26 It is S or R. In some embodiments, X 26 It is S. In some embodiments, X 26 It is R. In some embodiments, X 26 It is D.

[1119] In some embodiments, X 27 Not H. In some embodiments, X 27 It is H or L. In some embodiments, X 27 It is L. In some embodiments, X 27 It’s H.

[1120] In some embodiments,

[1121] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is M, X 14 It is I, X15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1122] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is M, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is I, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof;

[1123] 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, X 10 It is I, X 11 It is F, X 12 It is A, X 13 It is F, X 14 It is I, X 15 It is N, X 16 It is I, X 17 It is S, X 18 It is K, X 19 It is S, X 20 It is V, X 21 It is W, X 22 It is D, X 23 It is S, X 24 It is S, X 25 It is S, X 26 Is it S or X? 27 It is L, or a combination thereof; or

[1124] 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, X 10 It is I, X 11 It is F, X 12 It is S, X 13 It is M, X 14 It is V, X 15 It is Y, X 16 It is I, X 17 It is S, X 18 It is F, X 19 It is S, X 20 It is I, X 21 It is Y, X 22 It is V, X 23 It is S, X 24 It is A, X 25 It is S, X 26 Is it R or X? 27 It is L, or a combination thereof.

[1125] Constant structure domain

[1126] In some embodiments, the polypeptide (e.g., an antibody or an antigen-binding fragment thereof) comprises:

[1127] a) Antibody heavy chain constant domain sequence;

[1128] b) Antibody light chain constant domain sequence; or

[1129] Both a) and b)

[1130] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence. In some embodiments, the antibody heavy chain constant domain is selected from the group consisting of: IgA constant domain, IgD constant domain, IgE constant domain, IgG constant domain, and IgM constant domain.

[1131] In some embodiments, the IgG constant domain is an IgG1 constant domain (e.g., any one of SEQ ID NO: 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, an IgG2b, or an 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).

[1132] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:84).

[1133] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:85).

[1134] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:86).

[1135] ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:87).

[1136] ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHN AKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTFPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO:88).

[1137] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having at least about 60% sequence identity with an amino acid sequence of any one of SEQ ID NO:84-88 (e.g., any one of SEQ ID NO:84-86). For example, the polypeptide may contain 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 with any of the amino acid sequences in any of SEQ ID NO:84-88 (e.g., any of SEQ ID NO: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 with an amino acid sequence of any one of SEQ ID NO:84-88 (e.g., any one of SEQ ID NO:84-86).

[1138] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence containing at least one amino acid substitution (e.g., any one of SEQ ID NO: 84-86) relative to the amino acid sequence of any one of SEQ ID NO: 84-88. For example, the number of amino acid substitutions may 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 heavy chain constant domain sequence with about 1-10 amino acid substitutions (e.g., any one of SEQ ID NO: 84-86) relative to the amino acid sequence of any one of SEQ ID NO: 84-88.

[1139] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:84-88 (e.g., any one of SEQ ID NO:84-86).

[1140] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:84. 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 with 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 with the amino acid sequence of SEQ ID NO:84.

[1141] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising at least one amino acid substitution relative 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: 1-20, 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 heavy chain constant domain sequence comprising about 1-10 amino acid substitutions relative to the amino acid sequence of SEQ ID NO:84.

[1142] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having 100% sequence identity with the amino acid sequence of SEQ ID NO:84. The antibody disclosed herein using the name "AB-[]a" (e.g., AB-2a, see Table 4 for example) comprises the heavy chain constant domain amino acid sequence of SEQ ID NO:84, namely comprising M135, S137, and T139.

[1143] In some embodiments, the polypeptide comprises (e.g., an antibody or an antigen-binding fragment thereof) an antibody heavy chain constant domain sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:85. 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 with 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 with the amino acid sequence of SEQ ID NO: 85.

[1144] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence comprising at least one amino acid substitution relative 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: 1-20, 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 heavy chain constant domain sequence comprising about 1-10 amino acid substitutions relative to the amino acid sequence of SEQ ID NO:85.

[1145] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain sequence having 100% sequence identity with the amino acid sequence of SEQ ID NO:85. The antibody disclosed herein using the name "AB-[]b" (e.g., AB-2b, see, for example, Table 4) comprises the heavy chain constant domain amino acid sequence of SEQ ID NO:85, namely comprising Y135, T137, and E139.

[1146] 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) the 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, for example, Dumet et al., Insights into the IgG heavy chain engineering patent landscape as applied to IgG4 antibody development, Mabs. [Monoclonal Antibodies] 11(8):1341-50 (2019) (in particular Tables 1 and 2 therein) and WO02060919, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the Fc domain includes LS modification (i.e., substitution by M428L / N434S as determined by the Kabat number) or YTE modification (i.e., substitution by M252Y / S254T / T256E as determined by the Kabat number when comparing SEQ ID NO:84 and SEQ ID NO:85, corresponding to M135Y / S137T / T139E). In some embodiments, the Fc domain includes YTE modification.

[1147] In some embodiments, the polypeptide (e.g., an antibody or an antigen-binding fragment thereof) comprises a constant domain sequence of the antibody light chain.

[1148] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence selected from the group consisting of: a κ constant domain (e.g., SEQ ID NO: 89) and a λ constant domain (e.g., SEQ ID NO: 90).

[1149] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 89).

[1150] GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:90).

[1151] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:89 or SEQ ID NO:90. For example, the polypeptide may 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 with 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 with SEQ ID NO:89 or SEQ ID NO:90.

[1152] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence comprising at least one amino acid substitution relative 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 relative 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 conserved substitution. In some embodiments, at least one amino acid substitution is a highly conserved substitution.

[1153] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having 100% sequence identity with the amino acid sequence of SEQ ID NO:89 or SEQ ID NO:90.

[1154] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:90. For example, the polypeptide may 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 with 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 with SEQ ID NO:90.

[1155] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence comprising at least one amino acid substitution relative 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 relative to the amino acid sequence of SEQ ID NO:90. In some embodiments, at least one amino acid substitution is a conserved substitution. In some embodiments, at least one amino acid substitution is a highly conserved substitution.

[1156] In some embodiments, the polypeptide comprises an antibody light chain constant domain sequence having 100% sequence identity with the amino acid sequence of SEQ ID NO:90.

[1157] In some embodiments, the polypeptide comprises:

[1158] a) Antibody heavy chain constant domain sequence; and

[1159] b) Antibody light chain constant domain sequence.

[1160] In some embodiments, the polypeptide comprises an antibody heavy chain constant domain (e.g., SEQ ID NO:84 or SEQ ID NO:85) as an IgG1 constant domain and an antibody light chain constant domain (e.g., SEQ ID NO:90) as a λ constant domain.

[1161] In some embodiments, the polypeptide comprises:

[1162] a) An antibody heavy chain constant domain containing SEQ ID NO:85; or

[1163] b) Contains the antibody light chain constant domain of SEQ ID NO:90.

[1164] Or both a) and b).

[1165] In some embodiments, the polypeptide comprises:

[1166] a) An antibody heavy chain constant domain containing SEQ ID NO:85; and

[1167] b) Contains the antibody light chain constant domain of SEQ ID NO:90.

[1168] In some embodiments, the polypeptide comprises:

[1169] a) An antibody heavy chain constant domain containing SEQ ID NO:84; or

[1170] b) Contains the antibody light chain constant domain of SEQ ID NO:90.

[1171] Or both a) and b).

[1172] In some embodiments, the polypeptide comprises:

[1173] a) An antibody heavy chain constant domain containing SEQ ID NO:84; and

[1174] b) Contains the antibody light chain constant domain of SEQ ID NO:90.

[1175] Antibody and antigen binding fragment

[1176] In some embodiments, the polypeptide is an immunoglobulin molecule, such as an antibody (e.g., a whole antibody, an intact antibody) or an antigen-binding fragment of an antibody.

[1177] In some embodiments, the polypeptide is an antibody. As used herein, the term "antibody" refers to an immunoglobulin molecule capable of specifically binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site located in a variable domain of the immunoglobulin molecule. As used herein, the term "antibody" refers to a full-length antibody.

[1178] In some embodiments, the polypeptide is an antibody comprising two heavy (H) chains and two light (L) chains, or polymers thereof (e.g., IgM), linked by disulfide bonds. Each heavy chain contains V H And heavy chain constant structural domains (containing structural domain CH1, hinges CH2 and CH3). Each light chain contains V L And light chain constant structural domains (CL). V H and V L The region can be further subdivided into highly variable regions, called complementary determinant regions (CDRs), which are scattered within the frame regions (FRs). H and V L Each antibody contains three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The antibodies can be from any species, such as mouse antibodies, human antibodies, or humanized antibodies.

[1179] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of any one or more of SEQ ID NO:91-118. 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 with the amino acid sequence of any one or more of SEQ ID NO: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 with any one or more of the amino acid sequences in SEQ ID NO: 91-118.

[1180] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence that includes at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO: 91-118. For example, the number of amino acid substitutions may 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 that includes about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO: 91-118.

[1181] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:91-118.

[1182] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of any one or more of SEQ ID NO: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 with the amino acid sequence of any one or more of SEQ ID NO:91-94 and 105-108. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 80% sequence identity with an amino acid sequence of any one or more of SEQ ID NO: 91-94 and 105-108.

[1183] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence that includes at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:91-94 and 105-108. For example, the number of amino acid substitutions may 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 that includes about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO:91-94 and 105-108.

[1184] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO: 91-94 and 105-108.

[1185] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of any one or more of SEQ ID NO:91-94. 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 with the amino acid sequence of any one or more of SEQ ID NO: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 with an amino acid sequence of any one or more of SEQ ID NO:91-94.

[1186] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence that includes at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:91-94. For example, the number of amino acid substitutions may 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 that includes about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO:91-94.

[1187] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:91-94.

[1188] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:92. 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 with the amino acid sequence of SEQ ID NO:92. In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 70% or at least about 80% sequence identity with the amino acid sequence of SEQ ID NO:92.

[1189] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution relative 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 relative to the amino acid sequence of SEQ ID NO:92.

[1190] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity with the amino acid sequence of SEQ ID NO:92.

[1191] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of any one or more of SEQ ID NO: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 with the amino acid sequence of any one or more of SEQ ID NO: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 with any one or more of the amino acid sequences in SEQ ID NO:105-108.

[1192] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence that includes at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:105-108. For example, the number of amino acid substitutions may 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 that includes about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO:105-108.

[1193] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO:105-108.

[1194] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:106. 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 with 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 with the amino acid sequence of SEQ ID NO:106.

[1195] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence comprising at least one amino acid substitution relative 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 relative to the amino acid sequence of SEQ ID NO:106.

[1196] In some embodiments, the polypeptide comprises a heavy chain amino acid sequence having 100% sequence identity with the amino acid sequence of SEQ ID NO:106.

[1197] In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of any one or more of SEQ ID NO:119-129. For example, the polypeptide may 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 with the amino acid sequence of any one or more of SEQ ID NO: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 with any one or more of the amino acid sequences in SEQ ID NO:119-129.

[1198] In some embodiments, the polypeptide comprises a light chain amino acid sequence that includes at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:119-129. For example, the number of amino acid substitutions may 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 that includes about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO:119-129.

[1199] In some embodiments, the polypeptide comprises a light chain amino acid sequence having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:119-129.

[1200] In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of any one or more of SEQ ID NO:119-121. For example, the polypeptide may 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 with the amino acid sequence of any one or more of SEQ ID NO:119-121. In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 80% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:119-121.

[1201] In some embodiments, the polypeptide comprises a light chain amino acid sequence that includes at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO:119-121. For example, the number of amino acid substitutions may 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 that includes about 1-10 amino acid substitutions relative to any one or more of the amino acid sequences in SEQ ID NO:119-121.

[1202] In some embodiments, the polypeptide comprises a light chain amino acid sequence having 100% sequence identity with the amino acid sequence of any one of SEQ ID NO:119-121.

[1203] In some embodiments, the polypeptide comprises a light chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:120. For example, the polypeptide may 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 with 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 with the amino acid sequence of SEQ ID NO:120.

[1204] In some embodiments, the polypeptide comprises a light chain amino acid sequence comprising at least one amino acid substitution relative 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 relative to the amino acid sequence of SEQ ID NO:120.

[1205] In some embodiments, the polypeptide comprises a light chain amino acid sequence having 100% sequence identity with the amino acid sequence of SEQ ID NO:120.

[1206] In some embodiments, the polypeptide comprises:

[1207] a) A heavy chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 91-118; or

[1208] b) A light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 119-129, or

[1209] Both a) and b)

[1210] In some embodiments, the polypeptide comprises:

[1211] a) A heavy chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 91-118; and

[1212] b) A light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:119-129.

[1213] In some embodiments, the polypeptide comprises:

[1214] a) The heavy chain amino acid sequence contains at least one amino acid substituted amino acid relative to any one or more of the amino acid sequences in SEQ ID NO: 91-118; or

[1215] b) The amino acid sequence comprising at least one amino acid-substituted light chain amino acid sequence relative to any one or more of SEQ ID NO:119-129, or

[1216] Both a) and b)

[1217] In some embodiments, the polypeptide comprises:

[1218] a) A heavy chain amino acid sequence comprising at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO: 91-118; and

[1219] b) The amino acid sequence relative to any one or more of SEQ ID NO:119-129 contains a light chain amino acid sequence with at least one amino acid substitution.

[1220] In some embodiments, the polypeptide comprises:

[1221] a) A heavy chain amino acid sequence comprising about 1-10 substituted amino acids relative to any one or more of the amino acid sequences in SEQ ID NO: 91-118; or

[1222] b) A light chain amino acid sequence comprising about 1-10 amino acid substitutions relative to any one or more of SEQ ID NO: 119-129, or

[1223] Both a) and b)

[1224] In some embodiments, the polypeptide comprises:

[1225] a) A heavy chain amino acid sequence comprising about 1-10 substituted amino acids relative to any one or more of SEQ ID NO: 91-118; and

[1226] b) The amino acid sequence relative to any one or more of SEQ ID NO:119-129 contains a light chain amino acid sequence with about 1-10 amino acid substitutions.

[1227] In some embodiments, the polypeptide comprises:

[1228] a) A heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 91-118; or

[1229] b) A light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 119-129, or

[1230] Both a) and b)

[1231] In some embodiments, the polypeptide comprises:

[1232] a) A heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 91-118; and

[1233] b) A light chain amino acid sequence having 100% sequence identity with any of the amino acid sequences in SEQ ID NO:119-129.

[1234] In some embodiments, the polypeptide comprises:

[1235] a) A heavy chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 91-94 and 105-108; or

[1236] b) A light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 119-121, or

[1237] Both a) and b)

[1238] In some embodiments, the polypeptide comprises:

[1239] a) A heavy chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 91-94 and 105-108; and

[1240] b) A light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:119-121.

[1241] In some embodiments, the polypeptide comprises:

[1242] a) The amino acid sequence relative to any one or more of SEQ ID NO: 91-94 and 105-108 contains a heavy chain amino acid sequence with at least one amino acid substitution; or

[1243] b) The amino acid sequence comprising at least one amino acid-substituted light chain amino acid sequence relative to any one or more of SEQ ID NO:119-121, or

[1244] Both a) and b)

[1245] In some embodiments, the polypeptide comprises:

[1246] a) The amino acid sequence relative to any one or more of SEQ ID NO: 91-94 and 105-108 comprises a heavy chain amino acid sequence with at least one amino acid substitution; and

[1247] b) The amino acid sequence relative to any one or more of SEQ ID NO:119-121 contains a light chain amino acid sequence with at least one amino acid substitution.

[1248] In some embodiments, the polypeptide comprises:

[1249] a) The amino acid sequence relative to any one or more of SEQ ID NO: 91-94 and 105-108 comprises a heavy chain amino acid sequence with about 1-10 amino acid substitutions; or

[1250] b) A light chain amino acid sequence comprising about 1-10 amino acid substitutions relative to any one or more of SEQ ID NO: 119-121, or

[1251] Both a) and b)

[1252] In some embodiments, the polypeptide comprises:

[1253] a) A heavy chain amino acid sequence comprising about 1-10 substituted amino acids relative to any one or more of SEQ ID NO: 91-94 and 105-108; and

[1254] b) The amino acid sequence relative to any one or more of SEQ ID NO:119-121 contains a light chain amino acid sequence with about 1-10 amino acid substitutions.

[1255] In some embodiments, the polypeptide comprises:

[1256] a) A heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 91-94 and 105-108; or

[1257] b) A light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 119-121, or

[1258] Both a) and b)

[1259] In some embodiments, the polypeptide comprises:

[1260] a) A heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 91-94 and 105-108; and

[1261] b) A light chain amino acid sequence having 100% sequence identity with any of the amino acid sequences in SEQ ID NO:119-121.

[1262] In some embodiments, the polypeptide comprises:

[1263] a) A heavy chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 105-108; or

[1264] b) A light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 119-121, or

[1265] Both a) and b)

[1266] In some embodiments, the polypeptide comprises:

[1267] a) A heavy chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO: 105-108; and

[1268] b) A light chain amino acid sequence having at least about 60% sequence identity with any one or more of the amino acid sequences in SEQ ID NO:119-121.

[1269] In some embodiments, the polypeptide comprises:

[1270] a) The amino acid sequence relative to any one or more of SEQ ID NO: 105-108 contains a heavy chain amino acid sequence with at least one amino acid substitution; or

[1271] b) The amino acid sequence comprising at least one amino acid-substituted light chain amino acid sequence relative to any one or more of SEQ ID NO:119-121, or

[1272] Both a) and b)

[1273] In some embodiments, the polypeptide comprises:

[1274] a) A heavy chain amino acid sequence comprising at least one amino acid substitution relative to any one or more of the amino acid sequences in SEQ ID NO: 105-108; and

[1275] b) The amino acid sequence relative to any one or more of SEQ ID NO:119-121 contains a light chain amino acid sequence with at least one amino acid substitution.

[1276] In some embodiments, the polypeptide comprises:

[1277] a) A heavy chain amino acid sequence comprising about 1-10 substituted amino acids relative to any one or more of SEQ ID NO: 105-108; or

[1278] b) A light chain amino acid sequence comprising about 1-10 amino acid substitutions relative to any one or more of SEQ ID NO: 119-121, or

[1279] Both a) and b)

[1280] In some embodiments, the polypeptide comprises:

[1281] a) A heavy chain amino acid sequence comprising about 1-10 substituted amino acids relative to any one or more of SEQ ID NO: 105-108; and

[1282] b) The amino acid sequence relative to any one or more of SEQ ID NO:119-121 contains a light chain amino acid sequence with about 1-10 amino acid substitutions.

[1283] In some embodiments, the polypeptide comprises:

[1284] a) A heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 105-108; or

[1285] b) A light chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 119-121, or

[1286] Both a) and b)

[1287] In some embodiments, the polypeptide comprises:

[1288] a) A heavy chain amino acid sequence having 100% sequence identity with any one of the amino acid sequences in SEQ ID NO: 105-108; and

[1289] b) A light chain amino acid sequence having 100% sequence identity with any of the amino acid sequences in SEQ ID NO:119-121.

[1290] In some embodiments, the polypeptide comprises:

[1291] a) A heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:106; or

[1292] b) A light chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:120, or

[1293] Both a) and b)

[1294] In some embodiments, the polypeptide comprises:

[1295] a) A heavy chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:106; and

[1296] b) A light chain amino acid sequence having at least about 60% sequence identity with the amino acid sequence of SEQ ID NO:120.

[1297] In some embodiments, the polypeptide comprises:

[1298] a) The amino acid sequence relative to SEQ ID NO:106 contains a heavy chain amino acid sequence with at least one amino acid substitution; or

[1299] b) A light chain amino acid sequence containing approximately 1-10 substituted amino acids relative to the amino acid sequence of SEQ ID NO:120, or

[1300] Both a) and b)

[1301] In some embodiments, the polypeptide comprises:

[1302] a) A heavy chain amino acid sequence containing at least one amino acid substitution relative to the amino acid sequence of SEQ ID NO:106; and

[1303] b) The amino acid sequence relative to SEQ ID NO:120 contains a light chain amino acid sequence with about 1-10 amino acid substitutions.

[1304] In some embodiments, the polypeptide comprises:

[1305] a) A heavy chain containing the amino acid sequence of SEQ ID NO:105; and

[1306] b) A light chain comprising the amino acid sequence of SEQ ID NO:119(AB-1b). In some embodiments, the polypeptide comprises:

[1307] a) A heavy chain containing the amino acid sequence of SEQ ID NO:106; and

[1308] b) A light chain comprising the amino acid sequence of SEQ ID NO:120 (AB-2b). In some embodiments, the polypeptide comprises:

[1309] a) A heavy chain containing the amino acid sequence of SEQ ID NO:107; and

[1310] b) A light chain comprising the amino acid sequence of SEQ ID NO:119(AB-3b). In some embodiments, the polypeptide comprises:

[1311] a) A heavy chain containing the amino acid sequence of SEQ ID NO:108; and

[1312] b) A light chain comprising the amino acid sequence of SEQ ID NO:121 (AB-4b). In some embodiments, the polypeptide comprises:

[1313] a) A heavy chain containing the amino acid sequence of SEQ ID NO:109; and

[1314] b) A light chain comprising the amino acid sequence of SEQ ID NO:122 (AB-5b). In some embodiments, the polypeptide comprises:

[1315] a) A heavy chain containing the amino acid sequence of SEQ ID NO:110; and

[1316] b) A light chain comprising the amino acid sequence of SEQ ID NO:123 (AB-6b). In some embodiments, the polypeptide comprises:

[1317] a) A heavy chain containing the amino acid sequence of SEQ ID NO:111; and

[1318] b) A light chain comprising the amino acid sequence of SEQ ID NO:124 (AB-7b). In some embodiments, the polypeptide comprises:

[1319] a) A heavy chain containing the amino acid sequence of SEQ ID NO:112; and

[1320] b) A light chain comprising the amino acid sequence of SEQ ID NO:119 (AB-8b). In some embodiments, the polypeptide comprises:

[1321] a) A heavy chain containing the amino acid sequence of SEQ ID NO:113; and

[1322] b) A light chain comprising the amino acid sequence of SEQ ID NO:119(AB-9b). In some embodiments, the polypeptide comprises:

[1323] a) A heavy chain containing the amino acid sequence of SEQ ID NO:114; and

[1324] b) A light chain containing the amino acid sequence of SEQ ID NO:125(AB-10b).

[1325] In some embodiments, the polypeptide comprises:

[1326] a) A heavy chain containing the amino acid sequence of SEQ ID NO:115; and

[1327] b) A light chain containing the amino acid sequence of SEQ ID NO:126(AB-11b).

[1328] In some embodiments, the polypeptide comprises:

[1329] a) A heavy chain containing the amino acid sequence of SEQ ID NO:116; and

[1330] b) A light chain containing the amino acid sequence of SEQ ID NO:127(AB-12b).

[1331] In some embodiments, the polypeptide comprises:

[1332] a) A heavy chain containing the amino acid sequence of SEQ ID NO:117; and

[1333] b) A light chain containing the amino acid sequence of SEQ ID NO:128(AB-13b).

[1334] In some embodiments, the polypeptide comprises:

[1335] a) A heavy chain containing the amino acid sequence of SEQ ID NO:118; and

[1336] b) A light chain containing the amino acid sequence of SEQ ID NO:129(AB-14b).

[1337] 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 composed of a single monomeric variable antibody domain and capable of specifically binding to a target. Single-domain antibodies can be of any species, such as mouse antibodies, human antibodies, or humanized single-domain antibodies.

[1338] In some embodiments, VH domain and V L Domains can be linked together via adapters (e.g., synthetic adapters) to form various types of single-chain antibody designs, where V H / V L Domains pair within the molecule, or when V H and V L Domains that are expressed by independent chains pair in molecules to form monovalent antigen binding sites.

[1339] In some embodiments, the polypeptide is a heavy chain antibody or its antigen-binding fragment comprising two or more heavy chains but lacking light chains. Non-limiting examples of heavy chain antibodies include camel Vhh (also known as VHH or V). H H) Antibodies. Cameloid antibodies are antibodies from the camel family of mammals, including llamas, camels, and alpacas.

[1340] In some embodiments, the polypeptide is an antibody mimic. The term "antibody mimic" refers to a polypeptide that mimics the ability of an antibody to bind to an antigen, but is structurally different from the structure of a natural antibody. Non-limiting examples of antibody mimics include adnectin, affibodies, affilins, affimers, affitins, alphabodies, anticoagulants, avimers, DARPins, fynomers, Kunitz domain peptides, monomers, nanobodies, and nanoclamps.

[1341] 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 District, V L Region, Fab fragment, F(ab')2 fragment, Fd fragment, Fv fragment and a V H A structural domain or a V L Domain-specific antibodies (dAbs) composed of structural domains, etc. V H and V L The domains can be linked together via synthetic linkers to form various types of single-chain antibody designs, among which V H / V L Domains pair within the molecule, or when V H and V LIn cases where the domains are expressed by individual chains, they pair intermolecularly to form a monovalent antigen-binding site, such as a single-chain Fv (scFv) or a double-chain antibody. In some embodiments, the polypeptide disclosed herein is an antigen-binding fragment selected from Fab, F(ab')2, Fab', scFv, or Fv. In some embodiments, the polypeptide is scFv.

[1342] In some embodiments, the polypeptide is an isolated polypeptide.

[1343] In some embodiments, the polypeptide (e.g., isolated polypeptide) is generated by recombinant synthesis. In some embodiments, the polypeptide (e.g., isolated polypeptide) is generated by synthesis.

[1344] In some embodiments, the polypeptide is linked to a second polypeptide. The term "link" refers to attachment via covalent or non-covalent interactions. Conjugation can be performed using suitable linking agents. Non-limiting examples include peptide linkers, compound linkers, and chemical crosslinking agents. In some embodiments, the linker is a disulfide bond.

[1345] In some embodiments, the polypeptide is conjugated to a heterologous moiety. The term "conjugation" refers to attachment via covalent or non-covalent interactions. Conjugation can be performed using any one or more suitable linkers. Non-limiting examples include peptide linkers, compound linkers, and chemical crosslinking agents.

[1346] In some embodiments, the heterologous portion comprises a therapeutic agent, a diagnostic agent, or both. In some embodiments, the heterologous portion is selected from polyethylene glycol (PEG), hexadecanoic acid, hydrogels, nanoparticles, polymerized domains, and carrier peptides.

[1347] In some embodiments, the nanoparticles are lipid nanoparticles. In some embodiments, the nanoparticles are polymer 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) 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 polymer nanoparticles comprise poly(lactic acid-co-glycolic acid) acid (PLGA).

[1348] In some embodiments, the carrier peptide is albumin or Fc peptide.

[1349] In some embodiments, polypeptide:

[1350] a) Binds to TSLP protein, K D10 pM or lower (e.g., as measured by KinExA);

[1351] b) Binds to the AB loop region and C-terminal region of the D helix of the TSLP protein;

[1352] c) Binding to epitopes in TSLP proteins (e.g., full-length human TSLP);

[1353] d) Competes with the reference antibody to bind to the TSLP protein;

[1354] e) Reduce the binding of TSLP protein to TSLPR;

[1355] f) Modulate (e.g., reduce, inhibit, neutralize) TSLP-mediated biological activity, or

[1356] The aforementioned combination.

[1357] In some embodiments, the peptide is at 10 pM or lower K D (e.g., as measured by KinExA) combined with TSLP (e.g., human TSLP).

[1358] In some embodiments, the polypeptide binds (e.g., with binding affinity, with binding specificity) the AB loop region and C-terminal region of the helix D of the TSLP.

[1359] In some embodiments, the peptide has neutralizing activity against TSLP (e.g., full-length human TSLP). In some embodiments, the peptide has inhibitory activity against TSLP-mediated signaling.

[1360] In some embodiments, the peptide binds to a TSLP protein (e.g., SEQ ID NO:1), whose K D Approximately: 10pM, 8pM, 5pM, 4pM, 3pM, 2pM, 1.8pM, 1.5pM, 1.2pM, 1.0pM, 0.9pM, 0.8pM, 0.7pM, 0.6pM, 0.5pM, 0.4pM, 0.3pM, 0.2pM, or 0.1pM, or less; or approximately: 0.1-10pM, 0.2-10pM 0.2-8 pM, 0.3-8 pM, 0.3-5 pM, 0.4-5 pM, 0.4-4 pM, 0.5-4 pM, 0.5-3 pM, 0.6-3 pM, 0.6-2 pM, 0.7-2 pM, 0.7-1.8 pM, 0.8-1.8 pM, 0.8-1.5 pM, 1.0-1.5 pM, or 1.2-1.5 pM. In some embodiments, the peptide is at 2 pM or lower K. D Binds to TSLP proteins (e.g., SEQ ID NO:1).

[1361] In some embodiments, the peptide binds to a TSLP protein (e.g., SEQ ID NO:1), whose K D Approximately: 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 lower; or approximately: 0.1-2.0 pM, 0.2-2.0 pM, 0.2-1.9 pM. 0.3-1.9 pM, 0.3-1.8 pM, 0.4-1.8 pM, 0.4-1.7 pM, 0.5-1.7 pM, 0.5-1.6 pM, 0.6-1.6 pM, 0.6-1.5 pM, 0.7-1.5 pM, 0.7-1.4 pM, 0.8-1.4 pM, 0.8-1.3 pM, 0.9-1.3 pM, 1.0-1.3 pM, 1.0-1.2 pM, or 1.1-1.2 pM. In some embodiments, the peptide is at a Kc of about 1 pM or less. D Combine with TSLP.

[1362] In some embodiments, the polypeptide binds to the AB loop region and C-terminal region of the helix D of the TSLP protein.

[1363] In some embodiments, the peptide binds to an epitope in a TSLP protein (e.g., full-length human TSLP).

[1364] In some embodiments, the peptide competes with a reference antibody for binding to the TSLP protein (e.g., SEQ ID NO:1). Techniques and assays for assessing competition between antibodies are known in the art.

[1365] In some embodiments, the peptide reduces the 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 peptide reduces the binding of the TSLP protein to TSLPR, IC 50Approximately 500 pM or lower, such as approximately: 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 lower; or approximately: 0.1-500 pM, 0.1-450 pM 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.

[1366] In some embodiments, the peptide reduces the 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%, for example 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 peptide reduces the 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%.

[1367] In some embodiments, peptides regulate (e.g., reduce, inhibit, neutralize) TSLP-mediated biological activity.

[1368] Non-limiting examples of TSLP-mediated activity include TSLPR binding, TSLP-induced signal transducer and activator of transcription 5 (STAT5) signaling, TSLP-induced STAT3 signaling, TSLP-induced nuclear factor κ light chain enhancer (NFκB) signaling in activated B cells, TSLP-induced phosphoinositol 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, and IL-13, and mitogen-activated protein kinase kinase 2 (MEK2), as well as dendritic cell activation. In some embodiments, TSLP-mediated activity includes one or more innate immune mechanisms and / or one or more adaptive immune mechanisms.

[1369] TSLP-mediated activity can be determined, for example, by measuring STAT5 reporter activity, dendritic cell activation, TSLPR-expressing cell proliferation, 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. [Nature Communications] 8:14937 (2017), the contents of which are incorporated herein by reference).

[1370] In some embodiments, the peptide neutralizes the activity mediated by a TSLP protein (e.g., a full-length TSLP protein such as SEQ ID NO:1).

[1371] In some embodiments, the peptide neutralizes TSLP-mediated activity, IC 50 Approximately 500 pM or lower, such as approximately: 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 lower; or approximately: 0.1-500 pM, 0.1-450 pM 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.

[1372] In some embodiments, the peptide reduces TSLP-mediated activity by at least about 10%, for example, 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 peptide reduces TSLP-mediated activity by at least about 30%.

[1373] In some embodiments, the peptide reduces T2 inflammatory responses mediated by one or more innate immune mechanisms of TSLP.

[1374] Non-limiting examples of T2 inflammatory responses include activation of type 2 innate lymphocytes (ILC2); activation, migration, and / or local differentiation of eosinophil progenitor cells (EoP) (e.g., by upregulation of ICAM-1 and / or CD18, and / or by inhibition of L-selectin surface expression); increased eosinophil viability (e.g., by production of IL-6, eosinophil-derived neurotoxins, and chemokines (including CXC motif chemokine ligand 8 (CXCL8), CXCL1, and chemokine (CC motif) ligand 2 (CCL2)); basophil differentiation; mast cell production of T2 cytokines; mast cell production of chemokines CXCL8 and / or CCL1; and macrophage differentiation (e.g., by enhanced expression of CD80 activation markers).

[1375] In some embodiments, the peptide reduces the T2 inflammatory response mediated by one or more innate immune mechanisms of action of TSLP by at least about 10%, for example 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 peptide reduces the T2 inflammatory response mediated by one or more innate immune mechanisms of action of TSLP by at least about 30%.

[1376] In some embodiments, the peptide reduces T2 inflammatory responses mediated by one or more adaptive immune mechanisms of TSLP.

[1377] 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 co-stimulatory 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 (OX40 L); and increased expression of naïve T cell differentiation and / or polarization (e.g., naïve CD40 L). + T cells differentiate into TNF-α + IL-10 - T helper cells (Th2 cells); naïve CD4 + T cell proliferation and / or differentiation into Th2 cells or memory T cells; and CD8+ after TCR stimulation + Increased T cell proliferation.

[1378] In some embodiments, the peptide reduces the T2 inflammatory response mediated by the adaptive immune mechanism of TSLP by at least about 10%, for example 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 peptide reduces the T2 inflammatory response mediated by the adaptive immune mechanism of TSLP by at least about 30%.

[1379] In some embodiments, the peptide exhibits lower multispecificity for nonspecific targets (e.g., DNA) than a reference antibody. The DNA multispecific reagent (PSR) score is a measure of antibody binding to a nonspecific 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 multispecific binding activity, which is preferred for the peptides of the present invention.

[1380] In some embodiments, the polypeptide has a DNA PSR score of no more than 1, 2, 3, or 4. In some embodiments, the polypeptide has a DNA PSR score of 1, 2, 3, or 4. In some embodiments, the polypeptide has a DNA PSR score of 1-4, 1-3, 1-2, 2-4, 2-3, or 3-4. In some embodiments, the polypeptide has a DNA PSR score of 2-4, 2-3, or 3-4.

[1381] In some embodiments, the DNA PSR score of the peptide is at least about 10% lower than the DNA PSR score of the reference antibody, for example, 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 DNA PSR score of the peptide is at least about 30% lower than the DNA PSR score of the reference antibody.

[1382] In some embodiments, the DNA PSR score of the peptide is less than about 90% of the DNA PSR score of the reference antibody, for example, 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.

[1383] In some embodiments, the peptide has a DNA PSR score of about 1%-90% relative to the reference antibody's DNA PSR score, for example, about 2%-90%, 2%-85%, 3%-85%, 3%-80%, 4%-80%, 4%-75%, 5%-75%, 5%-70%, 6%-70%, 6%-65%, 7%-65%, 7%-60%, 8%-60%, 8%-55%, 9%-55%, 9%-50%, 10%-50%, 10%-45%, 15%-45%, 15%-40%, 20%-40%, 20%-35%, 25%-35%, or 25%-30% relative to the reference antibody.

[1384] In some embodiments, the peptide exhibits weaker self-association than a reference antibody, for example, as determined by affinity-capture self-interacting nanoparticle spectroscopy (AC-SINS) values. The AC-SINS value is the change in the maximum absorption wavelength of the absorption spectrum of the coated nanoparticle compared to the spectrum of the nanoparticle alone. Therefore, the greater the change in the maximum absorption wavelength, the stronger the self-interaction of the antibody coated on the nanoparticle. Self-association is an undesirable property, associated with poor viscosity and poor PK properties. Techniques and assays for assessing protein self-association are known in the art. See, for example, Patro & Przybycien, Biotechnol Bioeng. [Biotechnology and Bioengineering] 52(2):193-203 (1996), the contents of which are incorporated herein by reference in their entirety.

[1385] In some embodiments, the polypeptide has an AC-SINS value not exceeding the following: about 0, 1, 2, 3, 4, 5, 6, or 7. In some embodiments, the polypeptide has an AC-SINS value not exceeding the following: about 0, 1, 2, 3, or 4. In some embodiments, the polypeptide has the following AC-SINS values: about 0, 1, 2, 3, 4, 5, 6, or 7. In some embodiments, the polypeptide has the following AC-SINS values: about 0, 1, 2, 3, or 4. In some embodiments, the polypeptide has the following AC-SINS values: 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.

[1386] In some embodiments, the AC-SINS value of the peptide is at least about 10% lower than the AC-SINS value of the reference antibody, for example, 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 AC-SINS value of the peptide is at least about 30% lower than the AC-SINS value of the reference antibody.

[1387] In some embodiments, the AC-SINS value of the peptide is less than about 90% of the AC-SINS value of the reference antibody, for example, 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.

[1388] In some embodiments, the peptide has an AC-SINS value of about 1%-90% relative to the reference antibody, for example, about 2%-90%, 2%-85%, 3%-85%, 3%-80%, 4%-80%, 4%-75%, 5%-75%, 5%-70%, 6%-70%, 6%-65%, 7%-65%, 7%-60%, 8%-60%, 8%-55%, 9%-55%, 9%-50%, 10%-50%, 10%-45%, 15%-45%, 15%-40%, 20%-40%, 20%-35%, 25%-35%, or 25%-30% relative to the reference antibody.

[1389] Fusion protein

[1390] This disclosure provides, among other things, fusion proteins comprising one or more of the polypeptides disclosed herein.

[1391] The term "fusion protein" refers to a synthetic, semi-synthetic, or recombinant single protein molecule. Fusion proteins may comprise all or part of two or more distinct proteins and / or polypeptides linked by covalent bonds (e.g., peptide bonds).

[1392] The fusion proteins disclosed herein can be recombined or synthesized using conventional methods and reagents well known in the art. For example, the fusion proteins of the present invention can be recombined in suitable host cells (e.g., bacteria) according to methods known in the art. See, for example, Current Protocols in Molecular Biology, 2nd edition, edited by Ausubel et al., John Wiley & Sons, 1992; and Molecular Cloning: a Laboratory Manual, 2nd edition, Sambrook et al., 1989, Cold Spring Harbor Laboratory Press. For example, nucleic acid molecules containing the nucleotide sequence encoding the fusion protein described herein can be introduced and expressed in suitable host cells (e.g., *E. coli*), and the expressed fusions can be isolated / purified from the host cells (e.g., in inclusion bodies) using conventional methods and readily available reagents. For example, DNA fragments encoding different protein sequences (e.g., light-responsive domains, heteropeptide components) can be linked together according to conventional techniques. In another embodiment, the fusion gene can be synthesized using a variety of conventional techniques, including automated DNA synthesizers. Alternatively, PCR amplification of nucleic acid fragments can be performed using anchor primers that generate complementary overhangs between two consecutive nucleic acid fragments, which can then be annealed and re-amplified to produce a chimeric nucleic acid sequence (see Ausubel et al., Current Protocols in Molecular Biology, 1992).

[1393] Nucleic acids, vectors, host cells

[1394] This disclosure, among other things, provides one or more polynucleotides (e.g., DNA, RNA, or analogues thereof, optionally including one or more modified nucleotides; the polynucleotides may be linear or circular, such as linear or circular RNA). In some embodiments, the polypeptides or fusion proteins disclosed herein are encoded by a single polynucleotide. In some embodiments, the polypeptides or fusion proteins disclosed herein are encoded by multiple polynucleotides.

[1395] In some embodiments, the polynucleotide comprises a nucleotide sequence optimized for the selected host cell codon.

[1396] This disclosure also provides, among other things, vectors containing any one or more polynucleotides disclosed herein (e.g., expression vectors, including viral delivery vectors).

[1397] The term "expression vector" refers to a reproducible nucleic acid that can be used to express one or more proteins when transformed into a suitable expression host cell.

[1398] In some embodiments, the vector (e.g., an expression vector) further comprises an expression control polynucleotide sequence, a polynucleotide sequence encoding a selection marker, or both, operably linked to a polynucleotide. In some embodiments, the expression control polynucleotide sequence comprises a promoter sequence, an enhancer sequence, or both. In some embodiments, the expression control polynucleotide sequence comprises an inducible promoter sequence. The term "promoter" refers to a DNA region to which RNA polymerase binds and initiates gene transcription. The term "operably linked" means that the nucleic acid is located in a recombinant polynucleotide (e.g., a vector) in a manner that allows the nucleic acid to be expressed under the control of an element (e.g., a promoter) to which it is linked. The term "selection marker element" is an element that confers traits suitable for artificial selection. A selection marker element can be a negative selection marker or a positive selection marker.

[1399] This disclosure, among other things, provides expression host cells containing any one or more polynucleotides or expression vectors disclosed herein.

[1400] The term "expression host cell" refers to a cell that can be used to receive, maintain, replicate, and / or amplify 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 Escherichia coli, including DH5α, etc.

[1401] Composition

[1402] This disclosure provides, among other things, compositions comprising any of the polypeptides or fusion proteins disclosed herein. In some embodiments, the composition is a pharmaceutical composition.

[1403] In some embodiments, the composition (e.g., a pharmaceutical composition) further comprises a pharmaceutically acceptable carrier, excipient, stabilizer, diluent, or tonifier (Remington's Pharmaceutical Sciences 16th edition, Osol, A. ed. (1980)). A suitable pharmaceutically acceptable carrier, excipient, or stabilizer is non-toxic to the recipient at the dose and concentration employed. Non-limiting examples of pharmaceutically acceptable carriers, excipients, stabilizers, diluents, or tensioning agents include buffers (e.g., phosphates, citrates, histidines), 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 dextrin); 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); nonionic surfactants (e.g., Tween), and PLURONICS. TM And polyethylene glycol (PEG).

[1404] In some embodiments, the composition (e.g., a pharmaceutical composition) is formulated for a suitable schedule and route of administration. Non-limiting examples of routes of administration include oral, rectal, mucosal, intravenous, intramuscular, subcutaneous, and topical administration. In some embodiments, the composition (e.g., a pharmaceutical composition) is stored in the form of an aqueous solution or a dried formulation (e.g., lyophilized).

[1405] In some embodiments, the composition is formulated for administration by infusion (e.g., intravenous infusion). In other embodiments, the composition is formulated for subcutaneous administration.

[1406] In some embodiments, the composition is provided in a dosage form, for example, in a pre-filled syringe or an autoinjector.

[1407] In some embodiments, the pharmaceutical composition comprises about 50 mg to about 300 mg of the polypeptide or fusion protein disclosed herein, such as 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 the polypeptide or fusion protein disclosed herein, such as about: 60-280 mg, 80-280 mg, 80-260 mg, 100-260 mg, 100-250 mg, 120-250 mg, 120-240 mg, 140-240 mg, 140-220 mg, 150-250 mg, 150-200 mg, 160-220 mg, 160-200 mg, or 180-200 mg.

[1408] In some embodiments, the pharmaceutical composition comprises about 50 mg / ml to about 200 mg / ml of the polypeptide or fusion protein disclosed herein, such as 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 the polypeptide or fusion protein disclosed herein. In some embodiments, the pharmaceutical composition comprises about 60 mg / ml to about 200 mg / ml of the polypeptide or fusion protein disclosed herein, such as 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.

[1409] In some embodiments, the composition is formulated for administration as a combination therapy together with a second therapeutic agent. In some embodiments, the second therapeutic agent comprises a corticosteroid, a β-agonist (e.g., a long-acting β-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, ciroxonide, fluticasone furoate, fluticasone propionate, mometasone, and mometasone furoate. In some embodiments, the combination therapy comprises a β-agonist (e.g., a long-acting β-agonist). Non-limiting examples of long-acting β-agonists include salbutamol sulfate, formoterol fumarate, salmeterol oxaliplatin, atorol tartrate, olodaterol, and combinations thereof. In some embodiments, the combination therapy comprises a muscarinic antagonist. In some embodiments, the combination therapy comprises an IL-4 and / or IL-13 antagonist (e.g., for the treatment of allergic and / or asthma conditions). Non-limiting examples of IL-4 and / or IL-13 antagonists include Pitrakinra, anti-IL-13 antibodies or antigen-binding fragments thereof, anti-IL-4 antibodies or antigen-binding fragments thereof, and anti-IL-4R antibodies or antigen-binding fragments 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 antigens thereof (e.g., Pascolizumab (SB 240683)), and anti-IL-4R antibodies or antigen-binding fragments thereof (e.g., Dupilumab)).

[1410] In some embodiments, the composition is formulated for administration together with an inhaled corticosteroid (ICS), a long-acting β2-agonist (LABA), a long-acting muscarinic antagonist (LAMA), or any combination thereof.

[1411] In some embodiments, the composition (e.g., a pharmaceutical composition) comprises a polypeptide, the polypeptide comprising:

[1412] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:5; and

[1413] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:21(AB-2).

[1414] In some embodiments, the composition (e.g., a pharmaceutical composition) comprises a polypeptide, the polypeptide comprising:

[1415] a)V H It contains the amino acid sequence of SEQ ID NO:5, and

[1416] b)V L It contains the amino acid sequence of SEQ ID NO:21(AB-2).

[1417] In some embodiments, the composition (e.g., a pharmaceutical composition) comprises a polypeptide, the polypeptide comprising:

[1418] a) A heavy chain containing the amino acid sequence of SEQ ID NO:106; and

[1419] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2b).

[1420] In some embodiments, the composition (e.g., a pharmaceutical composition) comprises a polypeptide, the polypeptide comprising:

[1421] a) A heavy chain containing the amino acid sequence of SEQ ID NO:92; and

[1422] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2a).

[1423] How to use

[1424] This disclosure also provides, among other things, a method for neutralizing TSLP proteins (e.g., full-length TSLP proteins) in a subject, the method comprising administering to the subject an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable carrier and any of the polypeptides or fusion proteins disclosed herein as active ingredients.

[1425] In some embodiments, neutralizing TSLP proteins in a subject comprises administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1426] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:5; and

[1427] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:21(AB-2).

[1428] In some embodiments, neutralizing TSLP proteins in a subject comprises administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1429] a)V H It contains the amino acid sequence of SEQ ID NO:5, and

[1430] b)V L It contains the amino acid sequence of SEQ ID NO:21(AB-2).

[1431] In some embodiments, neutralizing TSLP proteins in a subject comprises administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1432] a) A heavy chain containing the amino acid sequence of SEQ ID NO:106; and

[1433] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2b).

[1434] In some embodiments, neutralizing TSLP proteins in a subject comprises administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1435] a) A heavy chain containing the amino acid sequence of SEQ ID NO:92; and

[1436] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2a).

[1437] In some embodiments, the subject is a mammal. In some embodiments, the subject is a mammal selected from the group consisting of: dogs, cats, mice, rats, hamsters, guinea pigs, horses, pigs, sheep, cattle, chimpanzees, macaques, cynomolgus monkeys, and humans. In some embodiments, the subject is a primate. In some embodiments, the subject is a human.

[1438] In some embodiments, the subject is 18 years of age or older, for example, 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.

[1439] 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 two years of age or older, for example, 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.

[1440] In some embodiments, the subjects are pediatric patients. In some embodiments, the subjects are pediatric patients aged 12 years or older.

[1441] 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 more.

[1442] 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 TSLP expression dysregulation, such as TSLP overexpression). 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.

[1443] Non-limiting examples of TSLP-related diseases and / or conditions (e.g., diseases and / or conditions associated with TSLP expression dysregulation) include asthma, Alzheimer's disease (AD), allergic conjunctivitis, chronic obstructive pulmonary disease (COPD), chronic spontaneous urticaria (CSU), nasal polyposis, rheumatoid arthritis (RA), sinusitis (RS) (e.g., allergic sinusitis, severe chronic sinusitis (e.g., severe chronic sinusitis with nasal polyposis)), cancer, and food allergies. Non-limiting examples of TSLP-related cancers (e.g., cancers associated with TSLP expression dysregulation) include lymphoma, acute lymphoblastic 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 their variants)).

[1444] In some embodiments, the subject has an inflammatory disorder (e.g., allergic inflammatory disorder). In some embodiments, the subject has an inflammatory disease or condition of the lungs. 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.

[1445] In some embodiments, the subject has been pretreated with one or more therapeutic agents prior to administration of the composition (e.g., a peptide or a pharmaceutical composition disclosed herein).

[1446] In some embodiments, the subject has previously received a therapeutic or preventative agent. For example, a COPD patient may have received inhaled corticosteroids (ICS), long-acting β2-agonists (LABA), long-acting muscarinic antagonists (LAMA), or any combination thereof.

[1447] This disclosure also provides, among other things, a method for treating a subject with a TSLP-related disease or condition, the method comprising administering to the subject an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable carrier and any of the polypeptides or fusion proteins disclosed herein as active ingredients.

[1448] In some embodiments, treating a subject for a TSLP-related disease or condition includes administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1449] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:5; and

[1450] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:21(AB-2).

[1451] In some embodiments, treating a subject for a TSLP-related disease or condition includes administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1452] a)V H It contains the amino acid sequence of SEQ ID NO:5, and

[1453] b)V L It contains the amino acid sequence of SEQ ID NO:21(AB-2).

[1454] In some embodiments, treating a subject for a TSLP-related disease or condition includes administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1455] a) A heavy chain containing the amino acid sequence of SEQ ID NO:106; and

[1456] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2b).

[1457] In some embodiments, treating a subject for a TSLP-related disease or condition includes administering an effective amount of a pharmaceutical composition to the subject, the pharmaceutical composition comprising a pharmaceutically acceptable carrier and a peptide, the peptide comprising:

[1458] a) A heavy chain containing the amino acid sequence of SEQ ID NO:92; and

[1459] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2a).

[1460] 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 Alzheimer's disease (AD).

[1461] In some embodiments, treatment of TSLP-related diseases or conditions is a preventative therapy.

[1462] In some embodiments, the effective amount is sufficient to prevent the subject from developing TSLP-related diseases or conditions (e.g., diseases or conditions associated with TSLP expression dysregulation).

[1463] In some embodiments, the effective amount is sufficient to reduce TSLPR occupancy in the subject. In some embodiments, the effective amount is sufficient to reduce TSLPR occupancy by at least about 10%, for example 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 TSLPR occupancy rate by approximately 10-99%, such as approximately: 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%.

[1464] In some embodiments, the effective amount is sufficient to reduce the TSLP receptor complex in the subject. In some embodiments, the effective amount is sufficient to reduce the TSLP receptor complex by at least about 10%, for example 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 TSLP receptor complex by about 10-99%, for example, 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%.

[1465] In some embodiments, the effective amount is sufficient to reduce the activity of the TSLP protein in the subject. In some embodiments, the effective amount is sufficient to reduce the activity of the TSLP protein by at least about 10%, for example 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 activity of the TSLP protein by about 10-99%, for example, 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%.

[1466] In some embodiments, the effective amount is sufficient to reduce airway inflammation in the subject (e.g., the number and / or density of one or more types of submucosal inflammatory cells in the airways). In some embodiments, the effective amount is sufficient to reduce airway inflammation by at least about 10%, for example 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 airway inflammation by about 10-99%, such as 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%.

[1467] In some embodiments, the effective amount is sufficient to reduce airway remodeling (e.g., reticular basement membrane thickening, airway epithelial integrity) in the subject. In some embodiments, the effective amount is sufficient to reduce airway remodeling by at least about 10%, for example 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 airway remodeling by about 10-99%, such as 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%.

[1468] In some embodiments, the effective amount is sufficient to treat the subject's nasal polyps (e.g., reduce the size and / or number of nasal polyps). In some embodiments, the effective amount is sufficient to reduce the size of the nasal polyps by at least about 10%, for example 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 the nasal polyp by about 10-99%, for example, 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%, for example 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%, for example, 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%.

[1469] The therapeutic agents disclosed herein can be administered via a variety of routes of administration, including, for example, local, transdermal, parenteral (e.g., intra-arterial, intravenous, intramuscular, subcutaneous, intradermal injection), intravenous infusion, and inhalation (e.g., intrabronchial, intranasal, or oral inhalation, intranasal drops), depending on the compound and the specific disease to be treated.

[1470] According to the instructions, application can be local or systemic. The preferred mode of application may vary depending on the specific compound selected.

[1471] In some embodiments, the composition (e.g., a peptide or pharmaceutical composition) is administered to the subject as a single therapy.

[1472] In some embodiments, the composition (e.g., a peptide or pharmaceutical composition) is administered to the subject in combination with one or more other therapeutic agents (e.g., simultaneously or sequentially with one or more other therapeutic agents) or preventive agents (e.g., simultaneously or sequentially with one or more preventive agents).

[1473] In some embodiments, treatment of TSLP-related diseases or conditions further includes administering a therapeutically effective amount of one or more additional therapeutic agents to the subject concurrently with or after the administration of the composition (e.g., a peptide or pharmaceutical composition). In some embodiments, treatment of TSLP-related diseases or conditions further includes administering a therapeutically effective amount of one or more additional preventative agents to the subject before, concurrently with, or after the administration of the composition (e.g., a peptide or pharmaceutical composition).

[1474] Administration of two or more therapeutic agents includes co-administering the therapeutic agents in a substantially simultaneous manner, such as in the form of a combination of drugs. Alternatively, such administration covers co-administration in multiple containers or co-administration of each therapeutic agent in separate containers (e.g., capsules, powders, and liquids). This administration also covers using each type of therapeutic agent sequentially, at approximately the same time or at different times. The composition and the second therapeutic agent may be administered via the same route of administration or via different routes of administration.

[1475] This disclosure also provides, among other things, a method for reducing (e.g., inhibiting) the binding of TSLP to TSLPR in cells, the method comprising contacting the cells with an effective amount of any one or more polypeptides or fusion proteins disclosed herein.

[1476] In some embodiments, reducing (e.g., inhibiting) the binding of TSLP to TSLPR in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1477] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:5; and

[1478] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:21(AB-2).

[1479] In some embodiments, reducing (e.g., inhibiting) the binding of TSLP to TSLPR in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1480] a)V H It contains the amino acid sequence of SEQ ID NO:5, and

[1481] b)VL It contains the amino acid sequence of SEQ ID NO:21(AB-2).

[1482] In some embodiments, reducing (e.g., inhibiting) the binding of TSLP to TSLPR in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1483] a) A heavy chain containing the amino acid sequence of SEQ ID NO:106; and

[1484] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2b).

[1485] In some embodiments, reducing (e.g., inhibiting) the binding of TSLP to TSLPR in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1486] a) A heavy chain containing the amino acid sequence of SEQ ID NO:92; and

[1487] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2a).

[1488] This disclosure provides, among other things, a method for inhibiting TSLP-induced signaling in cells, which involves contacting cells with an effective amount of any of the polypeptides or fusion proteins disclosed herein.

[1489] In some embodiments, inhibiting TSLP-induced signaling in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1490] a)V H It contains the amino acid sequences HCDR1, HCDR2, and HCDR3 of SEQ ID NO:5; and

[1491] b)V L It contains the amino acid sequences LCDR1, LCDR2 and LCDR3 of SEQ ID NO:21(AB-2).

[1492] In some embodiments, inhibiting TSLP-induced signaling in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1493] a)V H It contains the amino acid sequence of SEQ ID NO:5, and

[1494] b)V L It contains the amino acid sequence of SEQ ID NO:21(AB-2).

[1495] In some embodiments, inhibiting TSLP-induced signaling in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1496] a) A heavy chain containing the amino acid sequence of SEQ ID NO:106; and

[1497] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2b).

[1498] In some embodiments, inhibiting TSLP-induced signaling in cells includes contacting the cells with an effective amount of a polypeptide comprising:

[1499] a) A heavy chain containing the amino acid sequence of SEQ ID NO:92; and

[1500] b) A light chain containing the amino acid sequence of SEQ ID NO:120(AB-2a).

[1501] In some embodiments, the cells are selected from the group consisting of: airway smooth muscle cells (ASMC), basophils, dendritic cells, eosinophils, type 2 innate lymphocytes (ILC2), hematopoietic progenitor cells, lymphocytes, macrophages, mast cells, and monocytes.

[1502] AB-2

[1503] This disclosure also provides, among other things, a polypeptide comprising:

[1504] a) HCDR 1 containing the amino acid sequence of SEQ ID NO:31, HCDR 2 containing the amino acid sequence of SEQ ID NO:35, and HCDR 3 containing the amino acid sequence of SEQ ID NO:50; and

[1505] b) LCDR 1 containing the amino acid sequence of SEQ ID NO:60, LCDR 2 containing the amino acid sequence of SEQ ID NO:67, and LCDR 3 containing the amino acid sequence of SEQ ID NO:71.

[1506] The polypeptide is an antibody or its antigen-binding fragment.

[1507] This disclosure also provides, among other things, a polypeptide comprising:

[1508] a) HCDR1 composed of the amino acid sequence of SEQ ID NO:31, HCDR2 composed of the amino acid sequence of SEQ ID NO:35, and HCDR3 composed of the amino acid sequence of SEQ ID NO:50; and

[1509] b) LCDR1 composed of the amino acid sequence of SEQ ID NO:60, LCDR2 composed of the amino acid sequence of SEQ ID NO:67, and LCDR3 composed of the amino acid sequence of SEQ ID NO:71.

[1510] The polypeptide is an antibody or its antigen-binding fragment.

[1511] The amino acid sequence of AB-2CDR, as determined by Kabat numbering, is:

[1512] HCDR1: TYGMH (SEQ ID NO: 130)

[1513] HCDR2:VVWYDGSYTHYADSVKG(SEQ ID NO:131)

[1514] HCDR3:SPQWEEIFEAMDI(SEQ ID NO:132)

[1515] LCDR1:GGNNIGSKSVH(SEQ ID NO:133)

[1516] LCDR2:DDSDRPS(SEQ ID NO:134)

[1517] LCDR3: QIWDSSSSLVV(SEQ ID NO:71)

[1518] This disclosure also provides, among other things, a polypeptide comprising:

[1519] a) HCDR 1 containing the amino acid sequence of SEQ ID NO:130, HCDR 2 containing the amino acid sequence of SEQ ID NO:131, and HCDR 3 containing the amino acid sequence of SEQ ID NO:132; and

[1520] b) LCDR 1 containing the amino acid sequence of SEQ ID NO:133, LCDR 2 containing the amino acid sequence of SEQ ID NO:134, and LCDR 3 containing the amino acid sequence of SEQ ID NO:71.

[1521] The polypeptide is an antibody or its antigen-binding fragment.

[1522] This disclosure also provides, among other things, a polypeptide comprising:

[1523] a) HCDR1 composed of the amino acid sequence of SEQ ID NO:130, HCDR2 composed of the amino acid sequence of SEQ ID NO:131, and HCDR3 composed of the amino acid sequence of SEQ ID NO:132; and

[1524] b) LCDR1 composed of the amino acid sequence of SEQ ID NO:133, LCDR2 composed of the amino acid sequence of SEQ ID NO:134, and LCDR3 composed of the amino acid sequence of SEQ ID NO:71.

[1525] The polypeptide is an antibody or its antigen-binding fragment.

[1526] The amino acid sequence of AB-2CDR, as determined by Chothia numbering, is:

[1527] HCDR1: GFTFRTY(SEQ ID NO:135)

[1528] HCDR2: WYDGSY (SEQ ID NO:136)

[1529] HCDR3:SPQWEEIFEAMDI(SEQ ID NO:132)

[1530] LCDR1:GGNNIGSKSVH(SEQ ID NO:133)

[1531] LCDR2:DDSDRPS(SEQ ID NO:134)

[1532] LCDR3: QIWDSSSSLVV(SEQ ID NO:71)

[1533] This disclosure also provides, among other things, a polypeptide comprising:

[1534] a) HCDR 1 containing the amino acid sequence of SEQ ID NO:135, HCDR 2 containing the amino acid sequence of SEQ ID NO:136, and HCDR 3 containing the amino acid sequence of SEQ ID NO:132; and

[1535] b) LCDR 1 containing the amino acid sequence of SEQ ID NO:133, LCDR 2 containing the amino acid sequence of SEQ ID NO:134, and LCDR 3 containing the amino acid sequence of SEQ ID NO:71.

[1536] The polypeptide is an antibody or its antigen-binding fragment.

[1537] This disclosure also provides, among other things, a polypeptide comprising:

[1538] a) HCDR1 composed of the amino acid sequence of SEQ ID NO:135, HCDR2 composed of the amino acid sequence of SEQ ID NO:136, and HCDR3 composed of the amino acid sequence of SEQ ID NO:132; and

[1539] b) LCDR1 composed of the amino acid sequence of SEQ ID NO:133, LCDR2 composed of the amino acid sequence of SEQ ID NO:134, and LCDR3 composed of the amino acid sequence of SEQ ID NO:71.

[1540] The polypeptide is an antibody or its antigen-binding fragment.

[1541] In some embodiments, the polypeptide comprises:

[1542] a)V H It is humanized, containing the human frame region, or a combination thereof;

[1543] b)V L It is humanized, containing the human frame region, or a combination thereof.

[1544] Or both a) and b).

[1545] In some embodiments, the polypeptide comprises:

[1546] a)V H It contains the amino acid sequence of SEQ ID NO:5;

[1547] b)V L It contains the amino acid sequence of SEQ ID NO:21.

[1548] Or both a) and b).

[1549] In some embodiments, the antigen-binding fragment includes a single-chain variable fragment (scFv) and a heavy-chain variable restructure domain (V). HH ), antigen-binding fragment (Fab), Fab' or F(ab')2.

[1550] In some embodiments, the polypeptide comprises:

[1551] a) Antibody heavy chain constant domain;

[1552] b) Constant structural domain of antibody light chain

[1553] Or both a) and b).

[1554] 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 contains one or more mutations that increase the serum half-life of the antibody or its antigen-binding fragment in humans. In some embodiments, relative to the wild-type human IgG constant domain, the antibody heavy chain constant domain contains amino acid substitutions at amino acid residues 252, 254, and 256, respectively, using tyrosine, threonine, and glutamic acid, wherein these amino acid residues are numbered according to the EU index in Kabat.

[1555] In some embodiments, the polypeptide comprises:

[1556] a) An antibody heavy chain (HC) containing the amino acid sequence of SEQ ID NO:92;

[1557] b) An antibody light chain (LC) containing the amino acid sequence of SEQ ID NO:120,

[1558] Or both a) and b).

[1559] In some embodiments, the polypeptide comprises:

[1560] a) An antibody heavy chain (HC) containing the amino acid sequence of SEQ ID NO:106;

[1561] b) An antibody light chain (LC) containing the amino acid sequence of SEQ ID NO:120,

[1562] Or both a) and b).

[1563] In some embodiments, the peptide specifically binds to TSLP.

[1564] The present invention also provides a composition comprising one or more polypeptides disclosed in the portion entitled "AB-2", and one or more pharmaceutical excipients, diluents or carriers.

[1565] In some embodiments, the composition further comprises one or more additional therapeutic agents. In some embodiments, one or more additional therapeutic agents include corticosteroids, β-agonists, muscarinic antagonists, anti-inflammatory agents, IL-4 and / or IL-13 antagonists (e.g., antibodies or antigen-binding fragments of antibodies targeting IL-13, IL-4, or IL-4R) or combinations thereof.

[1566] This disclosure also provides, among other things, a method of treating a subject in need, comprising administering an effective amount of the composition disclosed in the section entitled "AB-2".

[1567] In some embodiments, the subject has asthma, atopic dermatitis (AD), allergic conjunctivitis, chronic obstructive pulmonary disease (COPD), chronic spontaneous urticaria (CSU), rheumatoid arthritis (RA), sinusitis (RS), eosinophilic esophagitis (EE), or food allergy. 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 moderate or severe asthma.

[1568] In some embodiments, the method further includes administering to the subject an effective amount of a corticosteroid, a β-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 a combination thereof.

[1569] This disclosure provides, among other things, a method for reducing the binding of TSLP to TSLPR on subject cells, the method comprising contacting the cells with an effective amount of the composition disclosed in the section entitled "AB-2".

[1570] This disclosure, among other things, provides a fusion protein comprising the polypeptide disclosed in the section entitled “AB-2”.

[1571] This disclosure, among other things, provides a polynucleotide containing the nucleotide sequence of the polypeptide disclosed in the section entitled "AB-2".

[1572] This disclosure, among other things, provides host cells containing the polynucleotides disclosed in the section entitled “AB-2”.

[1573] This disclosure also provides, among other things, a method for preparing the polypeptide disclosed in the section entitled "AB-2", which involves culturing a host cell containing a nucleotide sequence encoding the polypeptide under conditions in which the polypeptide is expressed in a host cell.

[1574] Sequence computation definition and verification

[1575] The applicant has facilitated an understanding of the structural and functional interactions of anti-TSLP antibodies through computational design and experimental verification, and this document provides those skilled in the art with means to utilize this understanding. For example, this application provides an appendix listing computer programs (also referred to herein as Appendices A and B, respectively) that can be used to evaluate (score) a given sequence or generate sequences that score above a given threshold when evaluated. These appendices include the Potts model (generalized Ising model) of peptides provided by this invention. In some embodiments, the implementation of the Potts model assumes that amino acids interact with their nearest neighbors, but it is also possible to determine relationships with distant amino acids (e.g., beyond one or more positions, thus representing features of secondary and tertiary structures).

[1576] The Potts model is a second-order sequence model. In some embodiments, the Potts model is energy-based. The model is designed to output the energy of a specific configuration. Given a sequence provided to the energy-based model, the model can be trained to produce quantizations of any measurement. The Potts model is represented by a model weight table. This table includes first-order and second-order weights. In the first-order weights, the table includes the position of a residue, the residue (e.g., an amino acid), and a score associated with the residue at that position. In the second-order weights, the table includes two positions, two residues, and a score for the two residues at the corresponding positions. This score is either self-energy or energy score.

[1577] The computer program listing appendices, referred to as Appendix A (claim.py) and Appendix B (fit_model.etab), are available online at the corresponding 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 running Python scripts. Those skilled in the art will also recognize that Appendix B can be loaded via a script from Appendix A. When running the claim.py file, two sequences (or a combined sequence) can be input. In embodiments, the source code and model of Appendix A and Appendix B are further described below.

[1578] Figure 12AFigure 1000 illustrates the zero-order, first-order, and second-order models. The zero-order model 1002 is a set of disordered amino acids. Since the model is zero-order, the amino acids have no order or structure. Using a physical analogy, it can be thought of as a "bag" of amino acids, because the amino acids in the bag are jumbled and have no order. Next, the first-order model 1004 is a matrix representing the quantifications (e.g., peptide mass fingerprints (PMF)) at each residue of the protein sequence. Each row of the matrix is ​​the amino acid sequence, and each column is configured such that the sum of the quantifications for each residue in that column is 1. These first-order energies can be called h tensors, denoted below as the "first hash table." First-order terms in the model are considered to be located at a given position, and the model can assign multiple points for each specific position. Using paired terms, the model can compare the scores of any two given positions. In an embodiment, the model can output an "energy," which is a number, and lower numbers are better.

[1579] The second-order sequence model 1006, or Potts model, represents the energy of two residues found together and the energy associated with the first-order effect. The second-order energy of the two residues found together is considered the j tensor, while the energy associated with the first-order effect is called the h tensor. In other words, the second-order energy compares the energy between two positions in an amino acid sequence. Therefore, the second-order sequence can compare the energy between two positions in any amino acid sequence. Those skilled in the art will recognize that this can be repeated for multiple sequences, thereby rating all residue pairs of many similar sequences for similarity.

[1580] Figure 12B This is a flowchart 1050 illustrating an example of a process for determining a claimed sequence according to an embodiment of this disclosure. First, the process receives a given polypeptide sequence or sequence pair (1052). In different embodiments, the sequence pair may be a single polypeptide chain, or in some embodiments, a single polypeptide chain, such as scFV. Those skilled in the art will also recognize that, in embodiments, the model of Appendix B can be used to generate polypeptide sequences or sequence pairs for testing using the script of Appendix A. The script of Appendix A scores the sequences using the model weights provided in Appendix B (1054). The script then determines whether the score is above a predetermined threshold (although those skilled in the art will recognize that, using different scoring mechanisms, the score may be below the threshold) (1056). If not, the polypeptide sequence or sequence pair is not verified (1058). If yes, the polypeptide sequence or sequence pair is verified (1060).

[1581] The Potts models disclosed in this paper were generated from experimental validation of approximately 100 sequences to identify sequences with beneficial functions. Model coefficients are stored in the ASCII text file [fit_model.etab]. These sequences exhibited acceptable multispecific reactivity, self-aggregation tendency, and expression at the time of analysis. Furthermore, they all possessed biological functions determined by SEAP reporting assays. The validated models only declared sequences not reported in existing literature.

[1582] In some embodiments, a script is used to determine the sequence pairs that require protection under this model (e.g., V). H and V L —Although for clarity, in different embodiments the sequence pairs may be separate polypeptide chains, or in some embodiments, a single polypeptide chain, such as scFV, whether it is above a threshold when scoring. This script is called a computational binding optimization (CBO) script. Loading two hash tables from an existing file, the script extracts the first-order and second-order index / position values, residue values, and corresponding scores for each hash table. For the first-order table, the script extracts a position, a residue, and a score associated with the residue at that position. For the second-order table, the script extracts a first position, a second position, a first residue, a second residue, and scores associated with the first residue at the first position and the second residue at the second position.

[1583] Two sequences (V) H Sequence and V L The sequence is input into the script and then compared. The comparison confirms V. H Sequence and V L The sequences are the same length as one or more reference sequences, and a given portion of both sequences is identical to one or more reference sequences. Once confirmed, residues are extracted from the two input sequences at specified positions to form a pruned and tandem sequence. The positions used for residue extraction are where mutations were applied during model training. The positions used to confirm that a given portion of the two sequences is identical to one or more reference sequences are where mutations were not applied during model training. Once pruned to the positions used for training the model, the two input sequences are tandemly concatenated. Energy scores for the given tandem sequence and two hash tables are then calculated. The following steps describe some embodiments for calculating the energy scores.

[1584] First of all, let Specify the sequence to be scored, where σ i It is its i-th amino acid. Furthermore, let h denote the self-energy matrix (e.g., a first-order matrix) of the Potts model, such that h i (a) is the energy associated with amino acid a at position i. Finally, let J denote the pairwise energy matrix of the model (e.g., a second-order matrix) such that J ij(a,b) represents the pair energy associated with amino acids a and b located at positions i and j, respectively. Using this representation, the model sequence... The total energy is:

[1585]

[1586] Where N is the length of the sequence. Furthermore, This can represent the sequence that leads to the sequence. The sequence in which the i-th amino acid is replaced by amino acid a. Using this representation, the sequence in the model... The pseudo-likelihood can be calculated as:

[1587]

[1588] Where μ is the total number of possible amino acids. In some embodiments, μ is 20, but those skilled in the art will recognize that the value of μ can vary. Finally, The logarithm of the pseudolikelihood can be expressed as:

[1589]

[1590] Appendix A's Score_Sequence function calculates and returns the above values. The Score_Sequence function returns the log probability of a sequence occurring given the Potts model weights input to the script. If the score is greater than a threshold, -7.7 in this example, the sequence satisfies the required parameters (e.g., confirmed / verified by the Potts model) and the function returns true. This score represents the log probability. Otherwise, the function returns false, and the sequence pair is not confirmed as a model. When the function returns true, it indicates that the sequence pair is suitable for a given purpose (e.g., a purpose related to combining with human TSLP).

[1591] Those skilled in the art will recognize that other thresholds representing the logarithmic probability that a sequence can satisfy the desired parameters can be used. In some embodiments, the range of instances of thresholds 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 threshold increases toward zero, the sequence space scoring within these thresholds represents a smaller sequence space with a higher confidence level. In some embodiments, the constants can be modified in the script or hash table, for example, by multiplying all constants by a constant. In this case, the threshold can be adjusted as can be determined by those skilled in the art. The threshold can also be calculated in a non-logarithmic manner and represented as a confidence interval, or as a non-logarithmic range, or as other probability thresholds.

[1592] In some embodiments, the script in Appendix A defines a sequence scoring function (sequence_energy). This function uses a Potts model to calculate the sequence energy of replacing residues (e.g., amino acids) with each possible residue at a set of positions in the sequence, the Potts model being trained to replace residues at those positions. As described above, the scoring checks each possible residue at each requested position. Those skilled in the art will recognize that this script can replace residues at each given position (as in the script in Appendix A). In some embodiments, a similar script can add residues to the sequence. Each score is calculated using the sequence_energy function and added to a score array. For each position checked, the scores in the array are summed and normalized using a softmax function. The scores are then logarithmically summed and returned.

[1593] In some embodiments, the sequence energy function takes a sequence as input and a hash table determined according to the input etab file (or another format). The function iterates over all first-order combinations, scoring them using a first hash table, and over all second-order combinations, scoring them using a second hash table. The sequence energy function considers all substituted residues and their first and second-order combinations, and sums these energies.

[1594] In some embodiments, the etab file is read using the `read_etab` function. Each line in the file with three items is unpacked and converted into an entry for a first-order combination in a first hash table. Each line in the file with five items is unpacked and converted into an entry for a second-order combination in a second hash table. Two hash tables are returned.

[1595] The steps and functions described above describe a "computationally optimized binding" (CBO) sequence, amino acid sequence, or peptide sequence. Such a sequence is one that returns a true value when input to the computationally optimized binding script described above. Those skilled in the art will recognize that this script can be implemented in other ways or through other series of steps. Those skilled in the art will also recognize that other functionally equivalent tables and constants can be used. However, the Python script, tables, and constants provided are configured, when executed by a processor, to output whether a given peptide sequence is a CBO sequence. Therefore, any sequence that results in a Boolean "true" output when the script runs is considered a CBO sequence. Any sequence that results in a Boolean "false" output when the script runs is considered excluded from the CBO sequence group. Thus, in some embodiments, the script in Appendix A does not itself determine a CBO sequence, but rather serves as a tool to confirm 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 called a CBO peptide.

[1596] In some embodiments, in addition to or separately from the Boolean value representing the threshold, similar scripts are able to return a calculated score for a given sequence or multiple sequences.

[1597] In some embodiments, a similar script can return one or more generated sequences from a Potts model. To generate 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 scoring function can be used to apply a brute-force method.

[1598] Given the above, Table 10 illustrates the pruned sequences (vlt, vht), tandem / combined sequences (vhvlt), and Potts model scores obtained when the script was applied to proteins AB-1 to AB-14. The sequences of AB-1 to AB-14 are shown in Tables 1 and 2. All proteins AB-1 to AB-14 have been or will be validated by the script, as their Potts model scores are higher than -7.7.

[1599] Based on Table 10, those skilled in the art will recognize that equivalent Potts model scores, such as scores with different scales or multiplied by a negative one, can result in scores equal to or higher than or lower than different thresholds.

[1600] Figure 13A computer network or similar digital processing environment in which embodiments of the invention can be implemented is shown.

[1601] Multiple client computers / devices 50 and multiple server computers 60 provide processing, storage, and input / output devices for executing applications, etc. The multiple client computers / devices 50 can also be linked to other computing devices, including other client devices / processes 50 and multiple server computers 60, via a communication network 70. The communication network 70 can be a remote access network, a global network (e.g., the Internet), a global computer collection, a local area network (LAN) or a wide area network (WAN), and can currently use their respective protocols (TCP / IP, etc.). Gateways for communication between devices (etc.). Other electronic device / computer network architectures are suitable.

[1602] Figure 14 yes Figure 13 This diagram illustrates an exemplary internal structure of a computer (e.g., client processor / device 50 or server computer 60) in a computer system. Each computer 50, 60 includes a system bus 79, where a bus is a set of hardware lines used for data transfer between components of a computer or processing system. The system bus 79 is essentially a shared conduit connecting different components of the computer system (e.g., processor, disk storage, memory, input / output ports, network ports, etc.), enabling the transfer of information between components. Connected to the system bus 79 is an I / O device interface 82 for connecting various input and output devices (e.g., keyboard, mouse, monitor, printer, speakers, etc.) to the computer 50, 60. A network interface 86 allows the computer to connect to a network (e.g., ...). Figure 13 Various other devices (network 70). Memory 90 provides volatile storage for computer software instructions 92 and data 94 (e.g., CBO determination scripts, Potts models, model weight code detailed above and in Appendices A and B) used to implement some embodiments of the invention. Disk storage 95 provides non-volatile storage for computer software instructions 92 and data 94 used to implement some embodiments of the invention. Central processing unit 84 is also attached to system bus 79 and provides execution of computer instructions.

[1603] In one embodiment, processor routine 92 and data 94 are computer program products (generally designated 92), referring to a non-transitory computer-readable medium (e.g., a removable storage medium, such as one or more DVD-ROMs, CD-ROMs, disks, magnetic tapes, etc.) that provides at least a portion of the software instructions for the system of the present invention. Computer program product 92 can be installed by any suitable software installer, as is known in the art. In another embodiment, at least a portion of the software instructions can also be downloaded via wired communication and / or wireless connection. In some embodiments, the program of the present invention is a computer program propagation signal product embodied on a propagation medium (e.g., radio waves, infrared waves, laser waves, sound waves, or radio waves propagating through a global network such as the Internet or one or more other networks). Such a carrier medium or signal can be used to provide at least a portion of the software instructions for the routine / program 92 of the present invention.

[1604] In another aspect, this disclosure provides peptides that specifically bind to TSLP (e.g., hTSLP): wherein the CBO model outputs a score above a predetermined threshold after scoring the amino acid sequence of the peptide.

[1605] In another aspect, this disclosure provides CBO peptides that specifically bind to TSLP (e.g., hTSLP): wherein the CBO model outputs a score above a predetermined threshold after scoring the amino acid sequence representing the CBO peptide.

[1606] On the other hand, this disclosure provides a polypeptide that specifically binds to TSLP (e.g., hTSLP) and obtains a score above a predetermined threshold from the CBO model after scoring the amino acid sequence representing the polypeptide.

[1607] In another aspect, this disclosure provides peptides that specifically bind to TSLP (e.g., hTSLP): wherein, after scoring the amino acid sequence representing the peptide, the peptide obtains a score above a predetermined threshold from the CBO model.

[1608] On the other hand, this disclosure provides computationally bound optimized (CBO) peptides determined by a computationally bound optimized (CBO) model that specifically binds to TSLP (e.g., hTSLP).

[1609] In some embodiments, the CBO peptide contains a complementary site substantially similar to the complementary site of an antibody comprising an amino acid sequence selected from:

[1610] SEQ ID NO:4 and SEQ ID NO:20(AB-1);

[1611] SEQ ID NO:5 and SEQ ID NO:21(AB-2);

[1612] SEQ ID NO:6 and SEQ ID NO:20(AB-3);

[1613] SEQ ID NO:7 and SEQ ID NO:22(AB-4);

[1614] SEQ ID NO:8 and SEQ ID NO:23(AB-5);

[1615] SEQ ID NO:9 and SEQ ID NO:24 (AB-6);

[1616] SEQ ID NO:10 and SEQ ID NO:25 (AB-7);

[1617] SEQ ID NO:11 and SEQ ID NO:20 (AB-8);

[1618] SEQ ID NO:12 and SEQ ID NO:20 (AB-9);

[1619] SEQ ID NO:13 and SEQ ID NO:26 (AB-10);

[1620] SEQ ID NO:14 and SEQ ID NO:27 (AB-11);

[1621] SEQ ID NO:15 and SEQ ID NO:28 (AB-12);

[1622] SEQ ID NO:16 and SEQ ID NO:29 (AB-13); or

[1623] SEQ ID NO:17 and SEQ ID NO:30 (AB-14), or

[1624] The aforementioned combination.

[1625] In some embodiments, the CBO polypeptide comprises a V selected from the following H / V LEpitopes of the combination that are substantially similar (e.g., having at least about 90% sequence identity; having 100% sequence identity): 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).

[1626] In some embodiments, the CBO polypeptide comprises a V selected from the following H / V LCombinations with different complementary positions: 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), by substitution of 1 to 3 (e.g., 1, 2 or 3) residues (e.g., conservative substitution, such as highly conservative substitution).

[1627] In certain embodiments, the CBO peptide does not contain V selected from the following H and V L right:

[1628] SEQ ID NO:4 and SEQ ID NO:20(AB-1),

[1629] SEQ ID NO:5 and SEQ ID NO:21(AB-2),

[1630] SEQ ID NO:6 and SEQ ID NO:20(AB-3),

[1631] SEQ ID NO:7 and SEQ ID NO:22(AB-4),

[1632] SEQ ID NO:8 and SEQ ID NO:23(AB-5),

[1633] SEQ ID ...

Claims

1. A polypeptide comprising: a) immunoglobulin heavy chain variable domain (V H ) amino acid sequence comprising a heavy chain complementary determining region 1 (HCDR1), a heavy chain complementary determining region 2 (HCDR2) and a heavy chain complementary determining region 3 (HCDR3) that are substantially similar to the amino acid sequence 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; and b) Immunoglobulin light chain variable domain (V L ) amino acid sequence comprising LCDR1, LCDR2 and LCDR3 that are substantially similar to the light chain complementary determining region 1 (LCDR1), light chain complementary determining region 2 (LCDR2) and light chain complementary determining region 3 (LCDR3) 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, respectively, Wherein the polypeptide is an antibody or an antigen-binding fragment thereof.

2. The polypeptide according to claim 1, comprising a V selected from the group consisting of H and V L HCDR1, HCDR2 and HCDR3 and LCDR1, LCDR2 and LCDR3 of the antibody: 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).

3. The polypeptide according to claim 1, comprising a V selected from the group consisting of H and V L right: 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).

4. A polypeptide comprising: 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, Wherein the polypeptide is an antibody or an antigen-binding fragment thereof.

5. A method comprising an immunoglobulin heavy chain variable domain (V H ) and immunoglobulin light chain variable domain (V L ), the polypeptide comprising a paratope substantially similar to the paratope of an antibody comprising an amino acid sequence selected from the group consisting of: 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 The above combination, Wherein the polypeptide is an antibody or an antigen-binding fragment thereof.

6. A method comprising an immunoglobulin heavy chain variable domain (V H ) wherein the V H Comprising the amino acid sequence of SEQ ID NO:2, wherein: 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 foregoing, And wherein the polypeptide is an antibody or an antigen-binding fragment thereof.

7. A method comprising an immunoglobulin heavy chain variable domain (V H ) wherein the V H Comprising the amino acid sequence of SEQ ID NO:2, wherein: 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 It is S; X 13 is M or L; X 14 It is V, or any combination of the foregoing, And wherein the polypeptide is an antibody or an antigen-binding fragment thereof.

8. The polypeptide of claim 6 or 7, further comprising an immunoglobulin light chain variable domain (V L ), where V L Comprising 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.

9. The polypeptide of claim 8, wherein: X 15 It is Y; X 16 is I; X 17 It is R; X 18 is F or Y; X 19 is N; X 20 is I; X 21 It is Y; X 22 is V or S; X 23 is M or E; X 24 It 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.

10. The polypeptide of any one of the preceding claims, wherein the polypeptide specifically binds to thymic stromal lymphopoietin (TSLP).

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

12. The polypeptide of any one of the preceding claims, 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. The polypeptide of claim 12, wherein: 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).

14. A polypeptide as described in 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 its antigen-binding fragment in humans, optionally wherein 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, relative to the wild-type human IgG constant domain, wherein the amino acid residues are numbered according to the EU index as in Kabat.

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

85.

16. A fusion protein comprising the polypeptide according to any one of the preceding claims.

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

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

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

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

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

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

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

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

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

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

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

30. A method for preparing the polypeptide of any one of claims 1-15, the method comprising culturing a host cell comprising a nucleotide sequence encoding the polypeptide under conditions whereby the polypeptide is expressed in the host cell.

31. A computer-implemented method comprising: The functional properties of a polypeptide comprising an amino acid sequence related to modulating the activity of a target molecule are scored using a computational binding optimization (CBO) model, which: For each amino acid position of the amino acid sequence of the polypeptide, a plurality of energy scores are calculated, the energy scores being based on the amino acid at the given position in the amino acid sequence, Add each energy score calculated to the array, Produces a normalized sum of the energy scores for each location, and A score for the polypeptide is generated by summing each of the calculated energy scores, which score represents a functional property of the polypeptide that is associated with the polypeptide modulating the activity of the target molecule.

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

33. The computer-implemented method of any one of claims 31-32, wherein scoring the polypeptide using the CBO model can be implemented by the script in Appendix A, and the CBO model is substantially similar to the table in Appendix B.

34. The method of any one of claims 31-33, wherein the functional characteristic is at least one of: Binding affinity for thymic stromal lymphopoietin (TSLP) polypeptide, characterized in that K D is 10 pM or less; (optionally, as measured by KinExA); Binding specificity to the AB loop region and 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 signaling, or a combination of the foregoing.

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

36. A system comprising: Processor; and A memory having computer code instructions stored thereon, the processor and the memory having the computer code instructions being configured to cause the system to: The functional properties of a polypeptide comprising an amino acid sequence related to regulating the activity of a target molecule are scored using the CBO model, which: For each amino acid position of the amino acid sequence of the polypeptide, a plurality of energy scores are calculated, the energy scores being based on the amino acid at the given position in the amino acid sequence, Add each energy score calculated to the array, Produces a normalized sum of the energy scores for each location, and A score for the amino acid sequence is generated by summing each of the calculated energy scores, which score represents the functional property of the polypeptide that is associated with the polypeptide modulating the activities of the target molecules.

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

38. The system of any one of claims 36-37, wherein scoring the polypeptide using the CBO model can be implemented by the script in Appendix A, and wherein the CBO model is substantially similar to the table in Appendix B.

39. The system of any one of claims 36-38, wherein the functional characteristic is at least one of the following: Binding affinity for thymic stromal lymphopoietin (TSLP) polypeptide, characterized in that K D is 10 pM or less; (optionally, as measured by KinExA); Binding specificity to the AB loop region and 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 signaling, or a combination of the foregoing.

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

41. A polypeptide that specifically binds to thymic stromal lymphopoietin (TSLP): The polypeptide comprises an amino acid sequence, and after scoring the amino acid sequence of the polypeptide, a score higher than a predetermined threshold is assigned to the amino acid sequence by a computational binding optimization (CBO) model.

42. The polypeptide of claim 41, wherein the polypeptide can be scored by the script of Computer Program Listing Appendix A, and wherein the CBO model is calculated by a table substantially similar to the table of Computer Program Listing Appendix B, and wherein the polypeptide is assigned a score calculated using the Computer Program Listing Appendix B; wherein the score is assigned to the polypeptide by a script substantially similar to that in Appendix A of the Computer Program Listing, which script employs a CBO model substantially similar to the table in Appendix B of the Computer Program Listing, wherein the log score of the polypeptide is 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. The polypeptide of any one of claims 41-42, wherein the polypeptide has one or more properties selected from the group consisting of: Binding affinity for thymic stromal lymphopoietin (TSLP) polypeptide, characterized in that K D is 10 pM or less; (optionally, as measured by KinExA); Binding specificity to the AB loop region and 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 signaling, or a combination of the foregoing.

44. The polypeptide of any one of claims 41-43, wherein the score output by the CBO model is equal to or higher than the score of the CBO model of one or more reference polypeptides, the reference polypeptide comprising a V selected from the group consisting of H and V L right: 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).

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

82.

46. ​​A polypeptide, The polypeptide comprises an amino acid sequence, and after scoring the amino acid sequence of the polypeptide, a score higher than a predetermined threshold is assigned to the amino acid sequence by a computational binding optimization (CBO) model.

47. A polypeptide that binds to human thymic stromal lymphopoietin (TSLP), wherein the polypeptide is designed by a method comprising: Generate peptide sequences using the CBO model; Use the CBO model to verify the generated peptide sequence: For each amino acid position in the polypeptide sequence, a plurality of energy scores are calculated based on replacing the amino acid at a given position in the polypeptide sequence with each of a plurality of different amino acids, Add each energy score calculated to the array, Generates the normalized sum of the energy scores for each position, Calculate the logarithm of each energy score calculated, and A score for the polypeptide sequence is generated by summing the logarithms of each energy score calculated, which score represents a functional property of the ability of the polypeptide to bind to human TSLP.

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