Antigen binding molecules targeting interleukin-4 receptor subunit alpha (il-4ra)

Abstract

The disclosure provides, in various embodiments, polypeptides (e.g., antibodies and antigen binding fragments thereof) that bind to IL-4 receptor subunit alpha (IL-4Rα). The disclosure also provides, in various embodiments, fusion proteins comprising one or more of polypeptides, polynucleotides encoding polypeptides, vectors and host cells suitable for expressing polypeptides, and methods for treating inflammatory conditions (e.g., atopic dermatitis).

Description

ANTIGEN BINDING MOLECULES TARGETING INTERLEUKIN-4 RECEPTOR SUBUNIT ALPHA (IL-4RA)RELATED APPLICATION

[0001] This Application claims the benefit of U.S. Provisional Application No. 63 / 659,671, filed on June 13, 2024. The entire teachings of the above applications are incorporated herein by reference.INCORPORATION BY REFERENCE OF MATERIAL IN XML

[0002] This application incorporates by reference the Sequence Listing contained in the following extensible Markup Language (XML) file being submitted concurrently herewith: a) File name: 57081079-002 Sequence Listing.xml; created June 13, 2025, 199,625 Bytes in size.BACKGROUND

[0003] The interleukin 4 (IL-4) / interleukin 13 (IL-13) axis has been implicated in numerous inflammatory conditions, including asthma, eczema (atopic dermatitis), food allergy, prurigo nodularis, chronic rhinosinusitis with nasal polyps, keloids, eosinophilic esophagitis, prostate cancer, chronic urticaria, bullous pemphigoid, localized scleroderma, alopecia areata, ulcerative colitis, aspirin-exacerbated respiratory disease, metastatic nonsmall cell lung cancer, and Netherton syndrome. According to the World Health Organization, in 2019 alone, an estimated 262 million people globally were affected by asthma, resulting in 455,000 deaths.

[0004] IL-4 receptor subunit alpha (TL-4Ru) mediates the activity of TL-4 and TL-13, two key proximal type 2 cytokines involved in type 2 immunity against mucosal pathogens and allergens (Gandhi et al., Targeting key proximal drivers of type 2 inflammation in disease, Nat. Rev. Drug Discov. 15:35-50 (2016)). Despite development of candidate anti-IL-4Ra therapies, only one has been FDA-approved. Accordingly, a widespread need exists for additional therapeutics that can target the IL-4Ra-mediated pathogenic pathways driving many immunological / inflammatory diseases (Nur Husna et al., IL-4 / IL-13 axis as therapeutic targets in allergic rhinitis and asthma, PeerJ. 10: 13444 (2022); Shi et al., Involvement oflL- 4, IL-13 and Their Receptors in Pancreatic Cancer, Int. J. Mol. Sci. 22(6):2998 (2021)).SUMMARY

[0005] There is a widespread need to develop novel therapeutic agents targeting the IL- 4 / IL-13 axis. The disclosure provides such therapeutics.

[0006] The disclosure provided herein is based, in part, on the discovery that polypeptides disclosed herein bind to interleukin-4 receptor alpha (IL-4Ra). Accordingly, the disclosure generally relates to compositions (e.g., polypeptides, pharmaceutical compositions) and methods that are useful for modulating (e.g., reducing) IL-4Ra-mediated immune activity.

[0007] The disclosure also provides, among other things, polypeptides (e.g., polypeptides that specifically binds an IL-4Ra) comprising: a VH amino acid sequence comprising a heavy chain complementarity determining region 1 (HCDR1), a heavy chain complementarity determining region 2 (HCDR2) and a heavy chain complementarity determining region 3 (HCDR3) that are substantially similar to a HCDR1, a HCDR2 and a HCDR3, respectively, of any one of SEQ ID NOs:6-20; and a VL amino acid sequence comprising a light chain complementarity determining region 1 (LCDR1), a light chain complementarity determining region 2 (LCDR2) and a light chain complementarity determining region 3 (LCDR3) that are substantially similar to a LCDR1, a LCDR2 and a LCDR3, respectively, of any one of SEQ ID NOs:25-39.

[0008] In some embodiments, a polypeptide comprises a HCDR1, HCDR2 and HCDR3, and a LCDR1, LCDR2 and LCDR3, of an antibody comprising a VH / VL combination selected from:SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); or SEQ ID NO:20 and SEQ ID NO:39 (AB-15c).

[0009] The disclosure provides, among other things, polypeptides (e.g., polypeptides that specifically binds an IL-4Ra) comprising a paratope that is substantially similar to a paratope of an antibody comprising an immunoglobulin heavy chain variable domain (Vn) / an immunoglobulin light chain variable domain (VL) combination selected from:SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); orSEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or a combination of any of the foregoing.

[0010] The disclosure also provides, among other things, polypeptides that comprise a VH comprising SEQ ID NO:4, wherein:Xi is not T;X2 is not R;X3 is not G;X4 is notN;X5 is not T;Xe is not K;X7 is not R;Xs is not L;Xg is not S;Xio is not I;Xu is not T;X12 is not I;X13 is not R;Xi4 is not R;X15 is not Y; orXi6 is not V; or any combination of the foregoing.

[0011] In some embodiments, a polypeptide comprises a VL comprising SEQ ID NO:23, wherein:X17 is not L;Xis is not I;X19 is not Y;X20 is not G;X21 is not A;X22 is not Q; orX23 is not T; or any combination of the foregoing.

[0012] In some embodiments, the disclosure provides polypeptides (e.g., polypeptides that specifically bind IL-4Ra) comprising: a VH sequence that has at least 70% sequence identity to SEQ ID NO:5; a VL sequence that has at least 70% sequence identity to SEQ ID NO:24; or a combination thereof, wherein the VH sequence does not comprise SEQ ID NO:5, the VL sequence does not comprise SEQ ID NO:24, or both.

[0013] In some embodiments, a polypeptide disclosed herein is a fusion protein.

[0014] In some embodiments, the disclosure provides polynucleotides encoding a polypeptide disclosed herein, vectors comprising such polynucleotides, and host cells comprising such polynucleotides and / or vectors.

[0015] In some embodiments, the disclosure provides compositions comprising a polypeptide, a fusion protein or a polynucleotide disclosed herein.

[0016] In some embodiments, the disclosure provides methods of treating a subject in need thereof, comprising administering an effective amount of the composition disclosed herein.

[0017] In some embodiments, the disclosure provides methods of reducing inflammation, a symptom of an inflammatory condition, or risk of developing an inflammatory condition in a cell of a subject, comprising contacting the cell with an effective amount of the composition disclosed herein.

[0018] An example embodiment is directed to a computer-implemented method for predicting a functional property of a polypeptide. The method begins by, via a computationally binding optimized (CBO) model, for each amino acid position of an amino acid sequence of the polypeptide: (1) determining a plurality of energy scores based on the amino acid position in the amino acid sequence, (2) generating a partition function based on the plurality of energy scores determined, and (3) determining a cross-entropy metric (e.g., cross-entropy loss) based on (i) an amino acid at the amino acid position in the amino acid sequence, (ii) a maximum energy score of the plurality of energy scores determined, and (iii) the generated partition function. An analysis score of the polypeptide is generated based on each cross-entropy metric determined. The analysis score indicates a predicted functional property of the polypeptide. According to some embodiments, the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:37. In some embodiments, the polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.

[0019] In some example embodiments, the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) amino acid sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least about 70% amino acid sequence identity (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) amino acid sequence identity to SEQ ID NO:37. In some example embodiments, the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising anamino acid sequence having at least 90% amino acid sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% amino acid sequence identity to SEQ ID NO:37. In some example embodiments, the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:37.

[0020] In an example embodiment, a given plurality of energy scores may include at least one of: a single amino acid energy score and a pairwise amino acid energy score.

[0021] According to an example embodiment, for at least one amino acid position of the amino acid sequence of the polypeptide, determining the plurality of energy scores may be further based on having substituted the amino acid at the amino acid position in the amino acid sequence with each of a plurality of different amino acids.

[0022] In an example embodiment, for at least one amino acid position of the amino acid sequence of the polypeptide, generating the partition function may be further based on a softmax function.

[0023] According to an example embodiment, at least one of: (1) predicting the functional property of the polypeptide may be implementable by a script of Appendix A or Appendix B, and (2) the CBO model may be substantially similar to a table of Appendix C.

[0024] In an example embodiment, the generated analysis score may be above a threshold. The threshold may be a score from the CBO model of one or more of a reference polypeptide that includes a VH and VL pair selected from:SEQ ID NO:5 and SEQ ID NO:24 (Reference);SEQ ID NO:6 and SEQ ID NO:25 (AB-1); SEQ ID NO:7 and SEQ ID NO:26 (AB-2); SEQ ID NO:8 and SEQ ID NO:27 (AB-3); SEQ ID NOV and SEQ ID NO:28 (AB-4); SEQ ID NO: 10 and SEQ ID NO:29 (AB-5); SEQ ID NO: 11 and SEQ ID NO:30 (AB-6); SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7); SEQ ID NO: 13 and SEQ ID NO:32 (AB-8); SEQ ID NO: 14 and SEQ ID NO:33 (AB-9); SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c);SEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or a combination of any of the foregoing.

[0025] According to an example embodiment, the predicted functional property may be a binding affinity for interleukin-4 receptor alpha (IL-4Ra).

[0026] In an example embodiment, the predicted functional property may be at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 1 pM or less, a binding affinity for IL-4Ra characterized by a kaof about 100 x 105M^s’1or less, a dissociation from IL-4Ra characterized by a kd of about 100 x 10’5s'1or less, a binding affinity for IL-4Ra characterized by an ECso of about 1 pM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 1 pM or less, a blocking activity against IL- 13 Type II signaling characterized by an ICso of about 1 pM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 1 pM or less.

[0027] According to an example embodiment, the predicted functional property may be at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.05 to 0.5 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7 x 105to 9 x 105T's-1or less, a dissociation from IL-4Ra characterized by a kd of about 2 x 10’5to 3 x 10’5s’ 1 or less, a binding affinity for IL-4Ra characterized by an EC50 of about 0.02 to 1.5 nM or less,a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 7.3 nM or less, a blocking activity against IL- 13 Type II signaling characterized by an IC50 of about 3 to 3.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 8 to 50 nM or less.

[0028] In an example embodiment, the predicted functional property may be at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.02 to 0.04 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7.7 x 105to 8.9 x 105M_|s_|or less, a dissociation from IL-4Ra characterized by a kd of about 2.1 x 10'5to 2.6 x 10'5s'1or less, a binding affinity for IL-4Ra characterized by an EC50 of about 0.04 to 1.3 nM or less, a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 4.4 to 7.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 30 to 50 nM or less.

[0029] According to an example embodiment, the predicted functional property may be at least one of: a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 1.8 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 30 to 34 nM or less.

[0030] In an example embodiment, the predicted functional property may be an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 8 to 8.8 nM or less.

[0031] According to an example embodiment, the predicted functional property may relate to modulating activity of a target molecule. The target molecule may be interleukin-4 receptor alpha (IL-4Ra).

[0032] In an example embodiment, the analysis score may be a binding score.

[0033] According to an example embodiment, the predicted functional property may be a binding affinity.

[0034] In an example embodiment, the polypeptide may not comprise a heavy chain having an amino acid sequence that is identical to SEQ ID NO: 104 and a light chain having an amino acid sequence that is identical to SEQ ID NO: 161.

[0035] Another example embodiment is directed to a computer-based system for predicting a functional property of a polypeptide. The system includes a processor and a memory with computer code instructions stored thereon. The processor and the memory, with the computer code instructions, are configured to cause the system to implement any embodiments or combination of embodiments described herein.

[0036] Yet another embodiment is directed to a computer program product for predicting a functional property of a polypeptide. The computer program product includes a non- transitory computer-readable medium with computer code instructions stored thereon. The computer code instructions are configured, when executed by a processor, to cause an apparatus associated with the processor to implement any embodiments or combination of embodiments described herein.

[0037] It is noted that embodiments of the method, system, and computer program product may be configured to implement any embodiments or combination of embodiments described herein.

[0038] An example embodiment is directed to a polypeptide that binds interleukin-4 receptor alpha (IL-4Ra). The polypeptide is assigned a score above a threshold by an analysis via a computationally binding optimized (CBO) model. The polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:37. The polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO: 5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.

[0039] In an example embodiment, at least one of: (1) the analysis may be implementable by a script of Appendix A or Appendix B employing the CBO model, (2) the CBO model may be calculated using a table substantially similar to that of Appendix C, (3) the score assigned to the polypeptide may be generated using Appendix C, (4) the polypeptide may beassigned the score by the script of Appendix A or Appendix B, and (5) the score assigned to the polypeptide may be at least about -27.8.

[0040] According to an example embodiment, the polypeptide may have at least one property selected from: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 1 pM or less, a binding affinity for IL-4Ra characterized by a kaof about 100 x 105M_|s-1or less, a dissociation from IL-4Ra characterized by a kd of about 100 x 10’5s'1or less, a binding affinity for IL-4Ra characterized by an ECso of about 1 pM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 1 pM or less, a blocking activity against IL- 13 Type II signaling characterized by an ICso of about 1 pM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 1 pM or less.

[0041] In an example embodiment, the polypeptide may have at least one property selected from: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.05 to 0.5 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7 x 105to 9 x 105T's-1or less, a dissociation from IL-4Ra characterized by a kd of about 2 x 10’5to 3 x 10’5s’ 1 or less, a binding affinity for IL-4Ra characterized by an EC50 of about 0.02 to 1.5 nM or less, a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 7.3 nM or less, a blocking activity against IL- 13 Type II signaling characterized by an IC50 of about 3 to 3.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by anIC50 of about 8 to 50 nM or less.

[0042] According to an example embodiment, the polypeptide may have at least one property selected from: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.02 to 0.04 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7.7 x 105to 8.9 x 105M_|s_|or less, a dissociation from IL-4Ra characterized by a kd of about 2.1 x 10'5to 2.6 x 10'5s'1or less, a binding affinity for IL-4Ra characterized by an ECso of about 0.04 to 1.3 nM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 4.4 to 7.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 30 to 50 nM or less.

[0043] In an example embodiment, the polypeptide has at least one property selected from: a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 1.8 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 30 to 34 nM or less.

[0044] According to an example embodiment, the polypeptide may have an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 8 to 8.8 nM or less.

[0045] In an example embodiment, the threshold may be a score from the CBO model of one or more of a reference polypeptide that includes a VH and VL pair selected from:SEQ ID NO:5 and SEQ ID NO:24 (Reference);SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c);SEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or a combination of any of the foregoing.

[0046] According to an example embodiment, the polypeptide may not comprise a heavy chain having an amino acid sequence that is identical to SEQ ID NO: 104 and a light chain having an amino acid sequence that is identical to SEQ ID NO: 161.

[0047] Another example embodiment is directed to a polypeptide that binds human interleukin-4 receptor alpha (IL-4Ra). The polypeptide is selected by a method comprising: evaluating a plurality of candidate polypeptides using a computationally binding optimized (CBO) model by, for each candidate polypeptide of the plurality of candidate polypeptides: for each amino acid position of an amino acid sequence of the candidate polypeptide: determining a plurality of energy scores based on the amino acid position in the amino acid sequence; generating a partition function based on the plurality of energy scores determined; and determining a cross-entropy metric based on (i) an amino acid at the amino acid position in the amino acid sequence, (ii) a maximum energy score of the plurality of energy scores determined, and (iii) the generated partition function; and generating an analysis score of the candidate polypeptide based on each cross-entropy metric determined, the analysis score indicating a functional property of the polypeptide’s ability to bind to human IL-4Ra; and selecting a given candidate polypeptide from among the plurality of candidate polypeptides based a result of the evaluating,wherein the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 37, and wherein the polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.BRIEF DESCRIPTION OF THE DRAWINGS

[0048] The foregoing will be apparent from the following more particular description of example embodiments, as illustrated in the accompanying drawings in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating embodiments.

[0049] In the drawings, “Reference” refers to the Reference Antibody.

[0050] FIG. 1 depicts the amino acid sequence of an epitope within the N-terminal receptor alpha domain of a human interleukin-4 receptor alpha (IL-4Ra) (an example sequence of IL-4Ra is set forth in SEQ ID NO: 1). The epitope residues bound by the Reference Antibody disclosed herein are indicated by asterisks (* ) . Also see non-limiting examples of IL-4Ra sequences (SEQ ID NOs: l-3) in Table 1.

[0051] FIGs. 2A and 2B depict an alignment of non-limiting examples of heavy chain variable domain (VH) amino acid sequences that are useful in polypeptides as disclosed herein. The heavy chain complementarity determining region (HCDR) amino acid sequences as determined by ImMunoGeneTics (IMGT) numbering (www.imgt.org / IMGTScientificChart / Nomenclature / IMGT-FRCDRdefmition.html, also accessible at www.imgt.org / ) are underlined. The bold letters indicate non-limiting examples of variable residues (designated throughout this disclosure by “Xn”) in the depicted sequences. An asterisk (*) indicates paratope residues. Paratope residues were defined as antibody residues in the Reference that, when bound to IL-4Ra, are within 5 angstroms (A) of the antigen. Also see VH consensus sequence (SEQ ID NO:4) for the Reference Antibody and AB-1 to AB-15 in Table 2. The antibody sequences were computationally generated using information from the sequence and structure of a reference polypeptide (“Reference”).

[0052] FIGs. 3 A and 3B depict an alignment of non-limiting examples of light chain variable domain (VL) amino acid sequences that are useful in polypeptides as disclosed herein. The light chain complementarity determining region (LCDR) amino acid sequences as determined by IMGT numbering are underlined. The bold letters indicate non-limiting examples of variable residues (designated throughout this disclosure by “Xn”) in the depicted sequences. An asterisk (*) indicates paratope residues. Paratope residues were defined as antibody residues in the Reference that, when bound to IL-4Ra, are within 5 angstroms (A) of the antigen. Also see VL consensus sequence (SEQ ID NO:23) for the Reference Antibody and AB-1 to AB- 15 in Table 2. The antibody sequences were computationally generated using information from the sequence and structure of a reference polypeptide (“Reference”).

[0053] FIGs. 4A-4C show antibody (mAb) binding in indirect ELISA binding assays. FIG. 4A shows hIgG4 ( “c” denotes wild-type IgG4) antibody binding as absorbance at 450 nm (A450) at varying antibody concentrations (nM). The Reference antibody is an IgG4 wild-type antibody. FIG. 4B shows hlgGl antibody binding as absorbance at 450 nm (A450) at varying antibody concentrations (nM). FIG. 4C shows hIgG4 (WT, YTE, or LS variant; “a” denotes IgG4 YTE variant, “b” denotes IgG4 LS variant, “c” denotes wild-type IgG4) antibody binding as absorbance at 450 nm (A450) at varying antibody concentrations (nM). The negative control was Isotype IgG4.

[0054] FIGs. 5A-5C show results from an assessment of the ability of antibodies to block IL-4-mediated activation of IL-4Ra via a HEK (human embryonic kidney)-Blue colorimetric assay (HEK-Blue (STAT6) assay). FIG. 5A shows hIgG4 (WT, YTE, or LS variant; “a” denotes IgG4 YTE variant, “b” denotes IgG4 LS variant, “c” denotes wild-type IgG4) antibody blocking as percent inhibition (% inhibition) at varying antibody (mAb) concentrations (nM). FIG. 5B shows hlgGl (“d” denotes IgGl) antibody blocking as percent inhibition at varying mAb concentrations (nM). FIG. 5C shows hIgG4 (WT, YTE, or LS variant) antibody blocking as percent inhibition at varying antibody mAb concentrations (nM). Isotype IgG4 was the negative control.

[0055] FIGs. 6A-6D show results from an IL-4Ra Human Primary Cell-based Assay (CD23 Inhibition assay) that used isolated primary B cells from four different donors. Percent inhibition was calculated from mean fluorescence intensity (MFI).

[0056] FIG. 7 shows results from a Ramos CD23 inhibition assay. Ramos cells were treated with an antibody or isotype control, then stained to identify CD23 inhibition by anti-IL-4Ra antibodies. Percent inhibition was calculated from mean fluorescence intensity (MFI).

[0057] FIGs. 8 A-8D depict results from blocking of IL-4 stimulation of SignalTransducer and Activator of Transcription 6 phosphorylation (pSTAT6) in whole human blood. The Reference and AB- 13c are IgG4 wild-type antibodies. FIGs. 8 A and 8C-8D show that with IL-4 stimulation (1 ng / mL), there is a dose-dependent decrease in pSTAT6 with both Reference Antibody (hIgG4) and AB-13c (hIgG4) in whole blood from three healthy donors in both B cells (CD19+) and T cells (CD4+) (Donor 1, FIG. 8 A; Donor 2, FIG. 8C; Donor 3, FIG. 8D). FIG. 8B shows representative images of a dose-dependent decrease in pSTAT6 expression in B and T cells in response to treatment with AB-13c or the Reference Antibody.

[0058] FIGs. 9A-9D depict evaluation of IL-4Ra target engagement in primary cells via functional inhibition of relevant atopic dermatitis (AD) biomarker Thymus- and Activation- Regulated Chemokine (TARC or CCL17). The Reference and AB-13c are IgG4 wild-type antibodies. FIG. 9A shows dose-dependent TARC stimulation seen with IL-4 in peripheral blood mononuclear cells (PBMCs) starting at 0.1 ng / mL of IL-4 with peak induction occurring at doses ^1 ng / mL. FIGs. 9B-9D show that AB-13c (hIgG4 WT) results in a dosedependent decrease of TARC release that is comparable to the Reference when cells are stimulated with 1 ng / mL (FIG. 9B), 10 ng / mL (FIG. 9C), or 100 ng / mL (FIG. 9D) of IL-4.

[0059] FIG. 10 is a flowchart of a method for predicting a functional property of a polypeptide according to an example embodiment.

[0060] FIG. 11 is a schematic view of a computer network in which embodiments may be implemented.

[0061] FIG. 12 is a block diagram illustrating an example embodiment of a computer node in the computer network of FIG. 11.DETAILED DESCRIPTION

[0062] A description of example embodiments follows.

[0063] Several aspects of the 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 the disclosure. One having ordinary skill in the relevant art, however, will readily recognize that the disclosure can be practiced without one or more of the specific details or practiced with other methods,protocols, reagents, cell lines and animals. The disclosure is not limited by the illustrated ordering of acts or events, as some acts may occur in different orders and / or concurrently with other acts or events. Furthermore, not all illustrated acts, steps or events are required to implement a methodology in accordance with the disclosure.Definitions

[0064] Unless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this disclosure pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and / or as otherwise defined herein.

[0065] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0066] When introducing elements disclosed herein, the articles “a,” “an,” “the,” and “said” are intended to mean that there are one or more of the elements. Further, the one or more elements may be the same or different. For example, unless the context clearly indicates otherwise, “a polypeptide” includes a single polypeptide, and two or more polypeptides.

[0067] Throughout this specification and the claims which follow, unless the context requires otherwise, the term “comprise,” and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of, e.g., a stated integer or step or group of integers or steps, but not the exclusion of any other integer or step or group of integer or step. When used herein, the term “comprising” can be substituted with the term “containing” or “including.”

[0068] As used herein, the term “consisting of’ excludes any element, step, or ingredient not specified in the claim element. When used herein, the term “consisting essentially of’ does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claim.

[0069] Any of the terms “comprising,” “containing,” “including,” and “having,” whenever used herein in the context of an aspect or embodiment disclosed herein, can insome embodiments, be replaced with the term “consisting of,” or “consisting essentially of’ to vary scopes disclosed herein.

[0070] As used herein, the conjunctive term “and / or” between multiple recited elements is understood as encompassing both individual and combined options. For instance, where two elements are conjoined by “and / or,” a first option refers to the applicability of the first element without the second. A second option refers to the applicability of the second element without the first. A third option refers to the applicability of the first and second elements together. Any one of these options is understood to fall within the meaning, and, therefore, satisfy the requirement of the term “and / or” as used herein. Concurrent applicability of more than one of the options is also understood to fall within the meaning, and, therefore, satisfy the requirement of the term “and / or.”

[0071] It should be understood that for all numerical bounds describing some parameter in this application, such as “about,” “at least,” “less than,” “fewer than,” and “more than,” the description also necessarily encompasses any range bounded by the recited values. Accordingly, for example, the description “at least 1, 2, 3, 4, or 5” also describes, inter alia, the ranges 1-2, 1-3, 1-4, 1-5, 2-3, 2-4, 2-5, 3-4, 3-5, and 4-5, et cetera.

[0072] When a list is presented, unless stated otherwise, it is to be understood that each individual element of that list, and every combination of that list, is a separate embodiment. For example, a list of embodiments presented as “A, B, or C” is to be interpreted as including the embodiments, “A,” “B,” “C,” “A or B,” “A or C,” “B or C,” or “A, B, or C .”

[0073] As used herein, the term “about” means within an acceptable error range for a particular value, as determined by one of ordinary skill in the art. Typically, an acceptable error range for a particular value depends, at least in part, on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within an acceptable standard deviation, per the practice in the art. Alternatively, “about” can mean a range of ± 20%, e.g., ± 10%, ± 5% or ± 1% of a given value. It is to be understood that the term “about” can precede any particular value specified herein, except for particular values used in the Exemplification. When “about” precedes a range, as in “90-99.9%,” the term “about” should be read as applying to both given values of the range, such that “about 90-99.9%” means about 90% to about 99.9%.

[0074] As used herein, the term “polypeptide” refers to a polymer of at least two amino acids covalently linked by an amide bond, regardless of length or post-translational modification (e.g., glycosylation or phosphorylation). A polypeptide can comprise anysuitable L-and / or D-amino acid, for example, common a-amino acids (e.g., alanine, glycine, valine), non-a-amino acids (e.g., [3-alanine, 4-aminobutyric acid, 6-aminocaproic acid, sarcosine, statine), and unusual amino acids (e.g., citrulline, homocitruline, homoserine, norleucine, norvaline, ornithine). The amino, carboxyl, and / or other functional groups on a polypeptide can be free (e.g., unmodified) or protected with a suitable protecting group. Suitable protecting groups for amino and carboxyl groups, and methods for adding or removing protecting groups are known in the art and are disclosed in, for example, Green and Wuts, “Protecting Groups in Organic Synthesis, ” John Wiley and Sons, 1991. The functional groups of a polypeptide can also be derivatized (e.g., alkylated) or labeled (e.g., with a detectable label, such as a fluorogen or a hapten) using methods known in the art. A polypeptide can comprise one or more modifications (e.g., amino acid linkers, acylation, acetylation, amidation, methylation, terminal modifiers (e.g., cyclizing modifications), N- methyl-a-amino group substitution), if desired. In addition, a polypeptide can be an analog of a known and / or naturally-occurring peptide, for example, a peptide analog having conservative amino acid residue substitution(s). The terms “polypeptide” and “protein” are used interchangeably herein.

[0075] 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, a polynucleotide disclosed herein comprises deoxyribonucleotides. In some embodiments, the polynucleotide comprises ribonucleotides. Non-limiting examples of polynucleotides include single-, double- or multi -stranded DNA or RNA, DNA-RNA hybrids (e.g., each “T” position may be independently substituted by a “U” or vice versa), or a polymer comprising purine and pyrimidine bases, or other natural, chemically, or biochemically modified, non-natural, or derivatized nucleotide bases. The backbone of the polynucleotide can comprise sugars and phosphate groups, modified or substituted sugar or phosphate groups, a polymer of synthetic subunits such as phosphoramidates, or a combination thereof.

[0076] As used herein, the term “sequence identity” refers to the extent to which two nucleotide sequences have the same residues at the same positions when the sequences are aligned to achieve a maximal level of identity, expressed as a percentage. For sequence alignment and comparison, typically one sequence is designated as a reference sequence, to which test sequences are compared. Sequence identity between reference and test sequences is expressed as a percentage of positions across the entire length of the reference sequence where the reference and test sequences share the same nucleotide or amino acid uponalignment of the reference and test sequences to achieve a maximal level of identity. As an example, two sequences are considered to have 70% sequence identity when, upon alignment to achieve a maximal level of identity, the test sequence has the same nucleotide residue at 70% of the same positions over the entire length of the reference sequence.

[0077] Alignment of sequences for comparison to achieve maximal levels of identity can be readily performed by a person of ordinary skill in the art using an appropriate alignment method or algorithm. In some instances, alignment can include introduced gaps to provide for the maximal level of identity. Examples include the local homology algorithm of Smith & Waterman, Adv. AppL Math. 2:482 (1981), the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), the search for similarity method of Pearson & Lipman, Proc. Nat’L Acad. Sci. USA 85:2444 (1988), computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), and visual inspection (see generally Ausubel et al., Current Protocols in Molecular Biology).

[0078] When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequent coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters. A commonly used tool for determining percent sequence identity is Protein Basic Local Alignment Search Tool (BLASTP) available through National Center for Biotechnology Information, National Library of Medicine, of the United States National Institutes of Health (Altschul etal., 1990).

[0079] As used herein, the term “substantially similar to” refers to a polypeptide disclosed herein that is substantially similar in amino acid sequence (e.g., has 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% of the amino acid residues amino acid sequence identity) and substantially preserves one or more functional properties of a specified polypeptide disclosed herein. In some embodiments, the one or more functional properties are selected from, without limitation, a substantially similar binding affinity, a substantially similar binding specificity, a substantially similar inhibitory activity, a substantially similar neutralization activity, and a substantially similar self-association property.

[0080] As used herein, a “complementarity determining region (CDR)” encompasses any CDR defined by an art-recognized method for identifying the CDR residues on an antibody.See, e.g., Kabat, E.A., et aL, (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242, Chothia et al., (1989) Nature 342:877; Chothia, C. et al., (1987) J. Mol. Biol. 196:901-917; Al-lazikani et al., (1997) J. 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. Two antibodies are determined to have the same CDR as one another with respect to a HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and / or LCDR3, when the identity of that CDR is determined for both antibodies using the same method.

[0081] The extent of the framework region and the CDRs of an antibody can be identified using one of several suitable methodologies that are well known in the art, for example, by the Kabat definition, the Chothia definition, the AbM definition, and / or the contact definition. Publicly and / or commercially available tools for identifying framework and / or CDR regions include, IgBlast (accessible at www.ncbi.nlm.nih.gov / igblast / ), Scaligner (available from drugdesigntech at www.scaligner.com / ), IMGT rules and / or tools (see, for example, www.imgt.org / IMGTScientificChart / Nomenclature / IMGT-FRCDRdefmition.html, also accessible at www.imgt.org / ), Chothia Canonical Assignment (accessible at www.bioinf.org.uk / abs / chothia.html), Antigen receptor Numbering And Receptor Calssificatilon (ANARCI, accessible at opig.stats.ox.ac.uk / webapps / newsabdab / sabpred / anarci / ), or see Vered Kunik, et al, Nucleic Acids Research, Volume 40, Issue Wl, 1 July 2012, Pages W521-W524).

[0082] As used herein, the term “antibody mimetic” refers to polypeptides capable of mimicking an antibody’s ability to bind an antigen, but structurally differ from native antibody structures. Examples of antibody mimetics include, but not limited to, Adnectins, Affibodies, Affilins, Affimers, Affitins, Alphabodies, Anticalins, Avimers, DARPins, Fynomers, Kunitz domain peptides, monobodies, nanobodies, nanoCLAMPs, and Versabodies.

[0083] The term “subject” refers to a mammalian subject, preferably human, diagnosed with or suspected of having an inflammatory condition in which the interleukin 4 (IL- 4) / interleukin 13 (IL- 13) axis has been implicated, such as, but not limited to: asthma, eczema (atopic dermatitis), food allergy, prurigo nodularis, chronic rhinosinusitis with nasal polyps, keloids, eosinophilic esophagitis, prostate cancer, chronic urticaria, bullous pemphigoid, localized scleroderma, alopecia areata, ulcerative colitis, aspirin-exacerbated respiratory disease, metastatic non-small cell lung cancer, and Netherton syndrome.

[0084] The phrase “pharmaceutically acceptable” means that the substance or composition the phrase modifies is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit / risk ratio.

[0085] As used herein, the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of mammals without undue toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, the relevant teachings of which are incorporated herein by reference in their entirety. Pharmaceutically acceptable salts of the agents / compounds described herein include salts derived from suitable inorganic and organic acids, and suitable inorganic and organic bases.

[0086] Examples of salts derived from suitable acids include salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts derived from suitable acids include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cinnamate, citrate, cyclopentanepropionate, di gluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, glutarate, glycolate, hemisulfate, heptanoate, hexanoate, hydroiodide, hydroxybenzoate, 2-hydroxy-ethanesulfonate, hydroxymaleate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 2-phenoxybenzoate, phenyl acetate, 3 -phenylpropionate, phosphate, pivalate, propionate, pyruvate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p- toluenesulfonate, undecanoate, valerate salts, and the like.

[0087] Either the mono-, di- or tri-acid salts can be formed, and such salts can exist in either a hydrated, solvated or substantially anhydrous form.

[0088] Salts derived from appropriate bases include salts derived from inorganic bases, such as alkali metal, alkaline earth metal, and ammonium bases, and salts derived from aliphatic, alicyclic or aromatic organic amines, such as methylamine, trimethylamine andpicoline, or N+((Ci-C4)alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, barium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.

[0089] “Pharmaceutically acceptable carrier” refers to a non-toxic carrier or excipient that does not destroy the pharmacological activity of the agent with which it is formulated and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the agent. Pharmaceutically acceptable carriers that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0090] “Treating” or “treatment,” as used herein, refers to taking steps to deliver a therapy to a subject, such as a mammal, in need thereof e.g., as by administering to a mammal one or more therapeutic agents). “Treating” or “treatment” includes inhibiting the disease or condition (e.g., as by slowing or stopping its progression or causing regression of the disease or condition) and relieving the symptoms resulting from the disease or condition. The term “treating,” or “treatment” refers to the medical management of a subject with the intent to improve, ameliorate, stabilize (i.e., not worsen), prevent, or cure a disease, pathological condition, or disorder — such as the particular indications exemplified herein. This term includes active treatment (treatment directed to improve the disease, pathological condition, or disorder), causal treatment (treatment directed to the cause of the associated disease, pathological condition, or disorder), palliative treatment (treatment designed for the relief of symptoms), preventative treatment (treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder); and supportive treatment (treatment employed to supplement another therapy). Treatment also includes diminishment of the extent of the disease or condition; preventing spread of the disease or condition; delay or slowing the progress of the disease or condition;amelioration or palliation of the disease or condition; and remission (whether partial or total), whether detectable or undetectable. “Ameliorating” or “palliating” a disease or condition means that the extent and / or undesirable clinical manifestations of the disease, disorder, or condition are lessened and / or time course of the progression is slowed or lengthened, as compared to the extent or time course in the absence of treatment. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the condition or disorder, as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.

[0091] A “pharmaceutical composition” refers to a formulation of one or more therapeutic agents and a medium generally accepted in the art for delivery of a biologically active agent to subjects, e.g., humans. In some embodiments, a pharmaceutical composition may include one or more pharmaceutically acceptable excipients, diluents, or carriers. In some embodiments, a pharmaceutical composition suitable for use in methods disclosed herein further comprises one or more pharmaceutically acceptable carriers.

[0092] “Pharmaceutically acceptable carrier, diluent, or excipient” includes any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.

[0093] “Pharmaceutically acceptable carrier” refers to an ingredient in a pharmaceutical composition, other than an active ingredient, which is nontoxic to a subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, excipient, stabilizer, or preservative. In some embodiments, the carrier may be a diluent, adjuvant, excipient, or vehicle with which the agent (e.g., polypeptide) is administered. Such vehicles may be liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. For example, 0.4% saline and 0.3% glycine can be used. These solutions are sterile and generally free of particulate matter. They may be sterilized by conventional, well-known sterilization techniques (e.g., filtration). The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, stabilizing, thickening, lubricating, and coloring agents, etc. The concentration of the agent in such pharmaceutical formulation may vary widely, z.e.,from less than about 0.5%, to at least about 1%, or to as much as 15% or 20%, 25%, 30%, 35%, 40%, 45% or 50% by weight. The concentration will be selected primarily based on required dose, fluid volumes, viscosities, etc., according to the mode of administration. Suitable vehicles and formulations, inclusive of other human proteins, e.g., human serum albumin, are described, for example, in Remington: The Science and Practice of Pharmacy, 21stEdition, Troy, D.B. ed., Lipincott Williams and Wilkins, Philadelphia, PA 2006, Part 5, Pharmaceutical Manufacturing: 691-1092 (e.g, pages 958-89).

[0094] Non-limiting examples of pharmaceutically acceptable carriers are solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible, such as salts, buffers, antioxidants, saccharides, aqueous or non-aqueous carriers, preservatives, wetting agents, surfactants or emulsifying agents, or combinations thereof.

[0095] Non-limiting examples of buffers are acetic acid, citric acid, formic acid, succinic acid, phosphoric acid, carbonic acid, malic acid, aspartic acid, histidine, boric acid, Tris buffers, HEPPSO, and HEPES.

[0096] Non-limiting examples of antioxidants are ascorbic acid, methionine, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, lecithin, citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, and tartaric acid.

[0097] Non-limiting examples of amino acids are histidine, isoleucine, methionine, glycine, arginine, lysine, L-leucine, tri-leucine, alanine, glutamic acid, L-threonine, and 2- phenylamine.

[0098] Non-limiting examples of surfactants are polysorbates (e.g, polysorbate-20 or polysorbate-80); poly oxamers (e.g., pol oxamer 188); Triton; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl- or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine (e.g., lauroamidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl- dimethylamine; sodium methyl cocoyl-, or disodium methyl oleyl-taurate; and the MONAQUA™ series (Mona Industries, Inc., Paterson, N.J.), poly ethyl glycol, polypropyl glycol, and copolymers of ethylene and propylene glycol (e.g., PLURONICS™, PF68, etc.).

[0099] Non-limiting examples of preservatives are phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrite, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride, alkylparaben (methyl, ethyl, propyl, butyl, and the like),benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof.

[0100] Non-limiting examples of saccharides are monosaccharides, di saccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, nonreducing sugars such as glucose, sucrose, trehalose, lactose, fructose, maltose, dextran, glycerin, dextran, erythritol, glycerol, arabitol, sylitol, sorbitol, mannitol, mellibiose, melezitose, raffinose, mannotriose, stachyose, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, or iso-maltulose.

[0101] Non-limiting examples of salts are acid addition salts and base addition salts. Acid addition salts include those derived from nontoxic inorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous, and the like, as well as from nontoxic organic acids such as aliphatic mono- and dicarboxylic acids, phenylsubstituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like. Base addition salts include those derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium, and the like, as well as from nontoxic organic amines, such as N,N’ -dibenzylethylenediamine, N-m ethyl glucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine, and the like. In some embodiments, the salt is sodium chloride (NaCl).

[0102] Agents (e.g., polypeptides) described herein may be prepared in accordance with standard procedures and are administered at dosages that are selected to reduce, prevent, or eliminate, or to slow or halt progression of, a condition being treated (see, e.g., Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, and Goodman and Gilman’s The Pharmaceutical Basis of Therapeutics, McGraw-Hill, New York, N.Y., the contents of which are incorporated herein by reference, for a general description of methods for administering various agents for human therapy).

[0103] “Administering” or “administration,” as used herein, refers to providing a compound, composition, or pharmaceutically acceptable salt thereof described herein to a subject in need of treatment or prevention. Administering can be performed, for example, once, a plurality of times, and / or over one or more extended periods. Administration includes both direct administration (including self-administration), and indirect administration (including an act of prescribing a drug or directing a subject to consume an agent). For example, as used herein, one (e.g., a physician) who instructs a subject (e.g., a human patient) to self-administer an agent (e.g., a drug), or to have an agent administered by another and / or who provides a patient with a prescription for a drug is administering an agent to a subject.

[0104] “A therapeutically effective amount” or “an effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic or biological result (e.g., treatment, healing, inhibition or amelioration of physiological response or condition, etc. . A therapeutically effective amount may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of a therapeutic or a combination of therapeutics to elicit a desired response in the individual.Polypeptides

[0105] Interleukin-4 Receptor Subunit Alpha (IL-4Ra)

[0106] As used herein, the terms “IL-4Ra” and “IL-4R” are used interchangeably. As used herein, IL-4Ra includes wild-type IL-4Ra proteins (e.g., wild-type human IL-4Ra proteins or homologs thereof) and subunits, domains (e.g., N-terminal cytokine homology domain (CHR) and ectodomain of Ra chain), and truncated forms thereof, mutant and engineered versions of full-length and truncated IL-4Ra proteins, and modified forms (e.g., post-translationally modified forms) of full-length and truncated IL-4Ra proteins. Nonlimiting examples of IL-4Ra sequences include SEQ ID NOs: l-3 (Table 1).

[0107] In some embodiments, a polypeptide binds to a wild-type IL-4Ra protein.

[0108] In some embodiments, a polypeptide binds to a mutant or engineered IL-4Ra protein. In some embodiments, a mutant or engineered IL-4Ra protein comprises an amino acid sequence having at least about 90% sequence identity to a wildtype IL-4Ra 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 to a wildtype IL-4Ra protein. In some embodiments, a mutant or engineered IL-4Ra protein comprises an amino acid sequence having about 90-99.9%, 90-99.8%, 92-99.8%, 92-99.6%, 94-99.6%, 94-99.5%, 95-99.5%, 95-99.4%, 96-99.4%, 96-99.2%, 97-99.2%, or 97-99% sequence identity to a wildtype IL-4Ra protein.

[0109] In some embodiments, a polypeptide binds to an IL-4Ra protein that comprises a mutation associated with a disease (e.g., severe asthma).

[0110] In some embodiments, a polypeptide binds to a modified IL-4Ra protein.

[0111] In some embodiments, a polypeptide is capable of binding to one or more epitope residues in an IL-4Ra protein, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 epitope residues of an IL-4Ra protein. In some embodiments, a polypeptide is capable of binding to one or more epitope residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, or all 17 residues) selected from Q38, L39, F41, L42, L43, S44, E45, H47, L64, D66, D67, V68, V69, A71, D72, N73, and Y74 of SEQ ID NO:1.

[0112] In some embodiments, a polypeptide binds to one or more epitope residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or all 17 residues) selected from Q38, L39, F41, L42, L43, S44, E45, H47, L64, D66, D67, V68, V69, A71, D72, N73, and Y74 of SEQ ID NO: 1.Comparator Polypeptides

[0113] As used herein, the term “comparator” or “comparator polypeptide” refers to a polypeptide (e.g., immunoglobulin molecule) that specifically binds to an IL-4Ra protein and is not a polypeptide disclosed herein. The sequence of a comparator polypeptide and a polypeptide disclosed herein may be compared to illustrate structural differences between them (e.g., differences at one or more amino acid positions, such as amino acid substitutions). Polypeptides disclosed herein have more than insubstantial differences (e.g., one or more substantial differences) in comparison to a comparator polypeptide, such that, polypeptides disclosed herein will, under controlled conditions, exhibit one or more (z.e., one, two, or all three) of: a different function, in a different way, to achieve a different result, in comparison to a comparator polypeptide. A comparator polypeptide may vary from a polypeptide disclosed herein by one or more amino acids, e.g., in some embodiments, by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acids. In some embodiments, a comparator polypeptide diverges from a polypeptide disclosed herein by at least about: 0.4%, 0.8%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or more amino acid percent identity.

[0114] In some embodiments, a comparator polypeptide comprises: a) a heavy chain complementarity determining region 1 (HCDR1), a heavy chain complementarity determining region 2 (HCDR2) and a heavy chain complementarity determining region 3 (HCDR3) sequences of SEQ ID NO:43, SEQ ID NO:47 and SEQ ID NO:55, respectively; b) a light chain complementarity determining region 1 (LCDR1), a light chain complementarity determining region 2 (LCDR2) and a light chain complementarity determining region 3 (LCDR3) sequences of SEQ ID NO:72, SEQ ID NO: 84 and SEQ ID NO: 87, respectively; or both a) and b).

[0115] In some embodiments, a comparator polypeptide comprises: a) a HCDR1, a HCDR2 and a HCDR3 sequences of SEQ ID NO:43, SEQ ID NO:47 and SEQ ID NO:55, respectively; and b) a LCDR1 , a LCDR2 and a LCDR3 sequences of SEQ ID NO : 72, SEQ ID NO:84 and SEQ ID NO:87, respectively.

[0116] See Table 3 and FIGs. 2A-2B for SEQ ID NOs:43, 47 and 55. See Table 3 andFIGs. 3A-3B for SEQ ID NOs:72, 84 and 87.

[0117] In some embodiments, a comparator polypeptide comprises: a) an immunoglobulin heavy chain variable region (VH) domain comprising the amino acid sequence of SEQ ID NO:5; b) an immunoglobulin light chain variable region (VL) domain comprising the amino acid sequence of SEQ ID NO:24; or both a) and b).

[0118] In some embodiments, a comparator polypeptide comprises: a) a VH domain comprising the amino acid sequence of SEQ ID NO: 5; and b) a VL domain comprising the amino acid sequence of SEQ ID NO:24.

[0119] See Table 2 and FIGs. 2A-2B for SEQ ID NO:5. See Table 2 and FIGs. 3A-3B for SEQ ID NO:24.

[0120] In some embodiments, a comparator polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 104; b) a light chain comprising the amino acid sequence of SEQ ID NO: 161; or both a) and b).

[0121] In some embodiments, a comparator polypeptide is an antibody, referred to herein as the “Reference Antibody” or “Reference.” The Reference Antibody comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 104; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 161.

[0122] See Table 5 for SEQ ID NO: 104. See Table 6 for SEQ ID NO: 161.

[0123] The Reference Antibody is an antibody that binds IL-4Ra protein and blocks IL-4 and IL- 13 activity.Variable Domains

[0124] In some embodiments, a polypeptide comprises an immunoglobulin heavy chain variable region (VH), an immunoglobulin light chain variable region (VL), or a VH and a VL.

[0125] In some embodiments, a polypeptide comprises a VH. In some embodiments, a polypeptide comprises a VH that is humanized, contains human framework regions, or both.

[0126] In some embodiments, a polypeptide comprises a VH having less than 100% sequence identity to the amino acid sequence of SEQ ID NO: 5.

[0127] In some embodiments, a polypeptide comprises a VH that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO:5. In some embodiments, a polypeptide comprises a VH that 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 to the amino acid sequence of SEQ ID NO:5. In some embodiments, a polypeptide comprises a VH that has at least about 85% sequence identity to the amino acid sequence of SEQ ID NO:5. In some embodiments, a polypeptide comprises a VH that has at least about 90% sequence identity to the amino acid sequence of SEQ ID NO: 5.

[0128] In some embodiments, a polypeptide comprises a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative substitution) relative to the amino acid sequence of SEQ ID NO:5. For example, the number of amino acid substitutions can be at least about: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about: 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4- 15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a VH that comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the at least one amino acid substitution replaces only a heavy chain complementarity determining region 1 (HCDR1), a heavy chain complementarity determining region 2 (HCDR2), and / or a heavy chain complementarity determining region 3 (HCDR3) residue, of SEQ ID NO:5. In some embodiments, the at least one amino acid substitution replaces only a non-CDR residue (e.g., within a framework region), of SEQ ID NO:5.

[0129] In some embodiments, an amino acid substitution is a conservative substitution. The term “a conservative amino acid substitution” or “a conservative substitution” refers to an amino acid substitution having a value of 0 or greater in BLOSUM62.

[0130] In some embodiments, an amino acid substitution is a highly conservative substitution. The term “a highly conservative amino acid substitution” or “a highly conservative substitution” refers to an amino acid substitution having a value of at least 1 (e.g., at least 2) in BLOSUM62.

[0131] In some embodiments, a polypeptide comprises a VH that has 100% sequence identity to the amino acid sequence of SEQ ID NO: 5. In some embodiments, a polypeptide comprises a VH that comprises the amino acid sequence of SEQ ID NO:5.

[0132] In some embodiments, a polypeptide comprises a VH that has at least about 70% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:6-20. The sequences identified as SEQ ID NOs:6-20 are shown in Table 2, which correspond to human VH domains. In some embodiments, a polypeptide comprises a VH that 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 to the amino acid sequence of any one or more of SEQ ID NOs:6-20. In some embodiments, a VH has at least about 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:6-20. In some embodiments, a polypeptide comprises a VH that has at least about 90% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:6-20.

[0133] In some embodiments, a polypeptide comprises a VH that comprises at least one amino acid substitution relative to the amino acid sequence of any one or more of SEQ ID NOs:6-20. For example, the number of amino acid substitutions can be at least about: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20, or about: 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a VH that comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of any one or more of SEQ ID NOs:6-20.

[0134] In some embodiments, a polypeptide comprises a VH that has 100% sequence identity to the amino acid sequence of any one of SEQ ID NO:6-20. In some embodiments, a polypeptide comprises a VH that comprises the amino acid sequence of any one of SEQ ID NO:6-20.

[0135] In some embodiments, a polypeptide comprises a VH that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, a polypeptide comprises a VH that 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 to the amino acid sequence of SEQ ID NO: 18. In some embodiments, a VH has at least about 85% sequence identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, a polypeptide comprises a VH that has at least about 90% sequence identity to the amino acid sequence of SEQ ID NO: 18.

[0136] In some embodiments, a polypeptide comprises a VH that comprises at least one amino acid substitution relative to the amino acid sequence of SEQ ID NO: 18. For example, the number of amino acid substitutions can be at least about: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20, or about: 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a VH that comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of SEQ ID NO: 18.

[0137] In some embodiments, a polypeptide comprises a VH that has 100% sequence identity to the amino acid sequence of SEQ ID NO: 18. In some embodiments, a polypeptide comprises a VH that comprises the amino acid sequence of SEQ ID NO:18.

[0138] In some embodiments, a polypeptide comprises a VL. In some embodiments, a polypeptide comprises a VL that is humanized, contains human framework regions, or both.

[0139] In some embodiments, a polypeptide comprises a VL having less than 100% sequence identity to the amino acid sequence of SEQ ID NO:24.

[0140] In some embodiments, a polypeptide comprises a VL that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO:24. In some embodiments, a polypeptide comprises a VL that 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 to the amino acid sequence of SEQ ID NO:24. In some embodiments, a polypeptide comprises a VL that has at least about 85% sequence identity to the amino acid sequence of SEQ ID NO:24. In some embodiments, a polypeptide comprises a VL that has at least about 90% sequence identity to the amino acid sequence of SEQ ID NO:24.

[0141] In some embodiments, a polypeptide comprises a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative substitution) relative to the amino acid sequence of SEQ ID NO:24. For example, the number of amino acid substitutions can be at least about: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about: 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4- 16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a VL that comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of SEQ ID NO:24. In some embodiments, the at least one amino acid substitution replaces only a light chain complementarity determining region 1 (LCDR1), a light chain complementarity determining region 2 (LCDR2), and / or a light chaincomplementarity determining region 3 (LCDR3) residue, of SEQ ID NO:24. In some embodiments, the at least one amino acid substitution replaces only a non-CDR residue (e.g., within a framework region), of SEQ ID NO:24.

[0142] In some embodiments, an amino acid substitution is a conservative substitution.

[0143] In some embodiments, an amino acid substitution is a highly conservative substitution.

[0144] In some embodiments, a polypeptide comprises a VL that has 100% sequence identity to the amino acid sequence of SEQ ID NO:24. In some embodiments, a polypeptide comprises a VL that comprises the amino acid sequence of SEQ ID NO:24.

[0145] In some embodiments, a polypeptide comprises a VL that has at least about 70% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:25-39. The sequences identified as SEQ ID NOs:25-39 are shown in Table 2, which correspond to human VL domains. In some embodiments, a polypeptide comprises a VL that 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 to the amino acid sequence of any one or more of SEQ ID NOs:25-39. In some embodiments, a VL has at least about 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:25-39. In some embodiments, a polypeptide comprises a VL that has at least about 90% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:25-39.

[0146] In some embodiments, a polypeptide comprises a VL that comprises at least one amino acid substitution relative to the amino acid sequence of any one or more of SEQ ID NOs:25-39. For example, the number of amino acid substitutions can be at least about: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about: 1-20, 1-19, 2-19, 2-18, 2- 17, 3-17, 3-16, 4-16, 4-15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a VL that comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of any one or more of SEQ ID NOs:25-39.

[0147] In some embodiments, a polypeptide comprises a VL that has 100% sequence identity to the amino acid sequence of any one of SEQ ID NO:25-39. In some embodiments, a polypeptide comprises a VL that comprises the amino acid sequence of any one of SEQ ID NO:25-39.

[0148] In some embodiments, a polypeptide comprises a VL that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO:37. In some embodiments, apolypeptide comprises a VL that 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 to the amino acid sequence of SEQ ID NO:37. In some embodiments, a VL has at least about 85% sequence identity to the amino acid sequence of SEQ ID NO:37. In some embodiments, a polypeptide comprises a VL that has at least about 90% sequence identity to the amino acid sequence of SEQ ID NO:37.

[0149] In some embodiments, a polypeptide comprises a VL that comprises at least one amino acid substitution relative to the amino acid sequence of SEQ ID NO:37. For example, the number of amino acid substitutions can be at least about: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or about: 1-20, 1-19, 2-19, 2-18, 2-17, 3-17, 3-16, 4-16, 4- 15, 5-15, 5-14, 6-14, 6-13, 7-13, 7-12, 8-12, 8-11, or 9-11. In some embodiments, a polypeptide comprises a VL that comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of SEQ ID NO:37.

[0150] In some embodiments, a polypeptide comprises a VL that has 100% sequence identity to the amino acid sequence of SEQ ID NO:37. In some embodiments, a polypeptide comprises a VL that comprises the amino acid sequence of SEQ ID NO:37.

[0151] In some embodiments, a polypeptide comprises a VH and a VL. In some embodiments, a polypeptide comprises a VH and VL that are humanized, contain human framework regions, or both.

[0152] In some embodiments, a polypeptide comprises: a) a VH having less than 100% sequence identity to the amino acid sequence of SEQ ID NO:5; b) a VL having less than 100% sequence identity to the amino acid sequence of SEQ ID NO:24; or both a) and b).

[0153] In some embodiments, a polypeptide comprises: a) a VH having less than 100% sequence identity to the amino acid sequence of SEQ ID NO: 5; and b) a VL having less than 100% sequence identity to the amino acid sequence of SEQ ID NO:24.

[0154] In some embodiments, a polypeptide comprises:a) a VH that has at least about 55% (e.g., at least about: 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:5; b) a VL that has at least about 55% (e.g., at least about: 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:24; or both a) and b), wherein the polypeptide does not comprise all 6 CDRs of an antibody comprising a VH amino acid sequence of SEQ ID NO: 5 and a VL amino acid sequence of SEQ ID NO:24.

[0155] In some embodiments, a polypeptide comprises: a) a VH that has at least about 55% (e.g., at least about: 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:5; and b) a VL that has at least about 55% (e.g., at least about: 60, 65, 70, 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:24, wherein the polypeptide does not comprise all 6 CDRs of an antibody comprising a VH amino acid sequence of SEQ ID NO: 5 and a VL amino acid sequence of SEQ ID NO:24.

[0156] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative substitution) relative to the amino acid sequence of SEQ ID NO: 5; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative substitution) relative to the amino acid sequence of SEQ ID NO:24; or both a) and b).

[0157] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of SEQ ID NO: 5; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of SEQ ID NO:24.

[0158] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a HCDR1, a HCDR2 and / or a HCDR3 residue, of SEQ ID NO:5; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a LCDR1, a LCDR2 and / or a LCDR3 residue, of SEQ ID NO:24; or both a) and b).

[0159] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a HCDR1, a HCDR2 and / or a HCDR3 residue, of SEQ ID NO:5; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a LCDR1, a LCDR2 and / or a LCDR3 residue, of SEQ ID NO:24.

[0160] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO: 5; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO:24; or both a) and b).

[0161] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO: 5; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO:24.

[0162] In some embodiments, a polypeptide comprises:a) a VH that has at least about 70% (e.g., at least about: 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:6-20; b) a VL that has at least about 70% (e.g., at least about: 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:25-39; or both a) and b).

[0163] In some embodiments, a polypeptide comprises: a) a VH that has at least about 70% (e.g., at least about: 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:6-20; and b) a VL that has at least about 70% (e.g., at least about: 75, 80, 85, 90, 95, 98, or 99%) sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:25-39.

[0164] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of any one or more of SEQ ID NOs:6-20; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of any one or more of SEQ ID NOs:25-39; or both a) and b).

[0165] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of any one or more of SEQ ID NOs:6-20; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of any one or more of SEQ ID NOs:25-39.

[0166] In some embodiments, a polypeptide comprises:a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a HCDR1, a HCDR2 and / or a HCDR3 residue, of any one or more of SEQ ID NOs: 6-20; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a LCDR1, a LCDR2 and / or a LCDR3 residue, of any one or more of SEQ ID NOs:25-39; or both a) and b).

[0167] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a HCDR1, a HCDR2 and / or a HCDR3 residue, of any one or more of SEQ ID NOs: 6-20; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a LCDR1, a LCDR2 and / or a LCDR3 residue, of any one or more of SEQ ID NOs:25-39.

[0168] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of any one or more of SEQ ID NOs: 6-20; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of any one or more of SEQ ID NOs:25-39; or both a) and b).

[0169] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of any one or more of SEQ ID NOs: 6-20; andb) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of any one or more of SEQ ID NOs:25-39.

[0170] In some embodiments, a polypeptide disclosed herein comprises: a) a VH comprising the amino acid sequence of any one of SEQ ID NOs:6-20; and b) a VL comprising the amino acid sequence of any one of SEQ ID NO:25-39.

[0171] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of any one of SEQ ID NOs:6-20; and b) a VL comprising the amino acid sequence of SEQ ID NO:24.

[0172] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO:5; and b) a VL comprising the amino acid sequence of any one of SEQ ID NO:25-39.

[0173] In some embodiments, a polypeptide comprises: a) a VH that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO: 18; b) a VL that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO:37; or both a) and b).

[0174] In some embodiments, a polypeptide comprises: a) a VH that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO: 18; and b) a VL that has at least about 70% sequence identity to the amino acid sequence of SEQ ID NO:37.

[0175] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of SEQ ID NO: 18; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of SEQ ID NO: 37; orboth a) and b).

[0176] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of SEQ ID NO: 18; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) relative to the amino acid sequence of SEQ ID NO:37.

[0177] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a HCDRl, a HCDR2 and / or a HCDR3 residue, of SEQ ID NO: 18; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a LCDR1, a LCDR2 and / or a LCDR3 residue, of SEQ ID NO:37; or both a) and b).

[0178] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a HCDRl, a HCDR2 and / or a HCDR3 residue, of SEQ ID NO: 18; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a LCDR1, a LCDR2 and / or a LCDR3 residue, of SEQ ID NO:37.

[0179] In some embodiments, a polypeptide comprises: a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO: 18; b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO:37; or both a) and b).

[0180] In some embodiments, a polypeptide comprises:a) a VH that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO: 18; and b) a VL that comprises at least one amino acid substitution (e.g., at least one conservative substitution such as highly conservative amino acid substitution) of a non-CDR residue (e.g., within a framework region), of SEQ ID NO:37.

[0181] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 18; and b) a VL comprising the amino acid sequence of any one of SEQ ID NO:24-39.

[0182] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of any one of SEQ ID NOs:5-20; and b) a VL comprising the amino acid sequence of SEQ ID NO:37.

[0183] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO:6; and b) a VL comprising the amino acid sequence of SEQ ID NO:25 (AB-1).

[0184] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO:7; and b) a VL comprising the amino acid sequence of SEQ ID NO:26 (AB-2).

[0185] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO:8; and b) a VL comprising the amino acid sequence of SEQ ID NO:27 (AB-3).

[0186] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NOV; and b) a VL comprising the amino acid sequence of SEQ ID NO:28 (AB-4).

[0187] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 10; and b) a VL comprising the amino acid sequence of SEQ ID NO:29 (AB-5).

[0188] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 11 ; and b) a VL comprising the amino acid sequence of SEQ ID NO:30 (AB-6).

[0189] In some embodiments, a polypeptide comprises:a) a VH comprising the amino acid sequence of SEQ ID NO: 12; and b) a VL comprising the amino acid sequence of SEQ ID NO:31 (AB-7).

[0190] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 13; and b) a VL comprising the amino acid sequence of SEQ ID NO:32 (AB-8).

[0191] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 14; and b) a VL comprising the amino acid sequence of SEQ ID NO:33 (AB-9).

[0192] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 15; and b) a VL comprising the amino acid sequence of SEQ ID NO:34 (AB-10).

[0193] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 16; and b) a VL comprising the amino acid sequence of SEQ ID NO:35 (AB-11).

[0194] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 17; and b) a VL comprising the amino acid sequence of SEQ ID NO:36 (AB-12).

[0195] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 18; and b) a VL comprising the amino acid sequence of SEQ ID NO:37 (AB-13).

[0196] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO: 19; and b) a VL comprising the amino acid sequence of SEQ ID NO:38 (AB-14).

[0197] In some embodiments, a polypeptide comprises: a) a VH comprising the amino acid sequence of SEQ ID NO:20; and b) a VL comprising the amino acid sequence of SEQ ID NO:39 (AB-15).Complementarity Determining Regions (CDRs)

[0198] A CDR (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and / or LCDR3) can be a CDR defined by any art-recognized method for identifying CDR residues of an antibody, as described further herein (e.g., a CDR as defined by Kabat, a CDR as defined by Chothia, or a CDR as defined by IMGT).

[0199] In some embodiments, a polypeptide does not comprise all six CDRs of an antibody comprising a VH amino acid sequence of SEQ ID NO:5 and a VL amino acid sequence of SEQ ID NO:24. In some embodiments, a polypeptide does not comprise all six sequences of SEQ ID NO:43, SEQ ID NO:47, SEQ ID NO:55, SEQ ID NO:72, SEQ ID NO:84 and SEQ ID NO:87. See Table 2 for SEQ ID NO:43, SEQ ID NO:47, SEQ ID NO:55, SEQ ID NO:72, SEQ ID NO:84 and SEQ ID NO:87.

[0200] In some embodiments, a polypeptide comprises fewer than six (e.g., 1, 2, 3, 4, or 5) CDRs of an antibody comprising a VH amino acid sequence of SEQ ID NO:5 and a VL amino acid sequence of SEQ ID NO:24. In some embodiments, a polypeptide comprises 1, 2, 3, 4, or 5 CDRs selected from SEQ ID NO:43, SEQ ID NO:47, SEQ ID NO:55, SEQ ID NO:72, SEQ ID NO:84 and SEQ ID NO:87.

[0201] In some embodiments, a polypeptide comprises all six CDRs of a specific polypeptide disclosed herein. In some embodiments, a polypeptide comprises fewer than six (e.g., 1, 2, 3, 4, or 5) of CDRs of a specific polypeptide disclosed herein.

[0202] In some embodiments, a polypeptide comprises: a) a VH amino acid sequence comprising a HCDR1, a HCDR2, and a HCDR3 that are substantially similar in amino acid sequence to a HCDR1, a HCDR2 and a HCDR3, respectively, of a VH amino acid sequence set forth in any one of SEQ ID NOs: 6-20; b) a VL amino acid sequence comprising a LCDR1, a LCDR2 and a LCDR3 that are substantially similar in amino acid sequence to a LCDR1, a LCDR2 and a LCDR3, respectively, of a VL amino acid sequence set forth in any one of SEQ ID NOs:25-39; or both a) and b).

[0203] In some embodiments, a polypeptide comprises: a) a VH amino acid sequence comprising a HCDR1, a HCDR2, and a HCDR3 that are substantially similar in amino acid sequence to a HCDR1, a HCDR2 and a HCDR3, respectively, of a VH amino acid sequence set forth in any one of SEQ ID NOs: 6-20; and b) a VL amino acid sequence comprising a LCDR1, a LCDR2 and a LCDR3 that are substantially similar in amino acid sequence to a LCDR1, a LCDR2 and a LCDR3, respectively, of a VL amino acid sequence set forth in any one of SEQ ID NOs:25-39.

[0204] See Tables 2-3 and FIGs. 2A-3B for SEQ ID NOs:6-20 and 25-39, and nonlimiting examples of corresponding HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences.

[0205] In some embodiments, a polypeptide comprises: a) a VH amino acid sequence comprising a HCDR1, a HCDR2, and a HCDR3 that are substantially similar in amino acid sequence to a HCDR1, a HCDR2 and a HCDR3, respectively, of a VH amino acid sequence of any one or more of SEQ ID NOs: 6-20; and b) a VL amino acid sequence comprising a LCDR1, a LCDR2, and a LCDR3 that are substantially similar in amino acid sequence to a LCDR1, a LCDR2 and a LCDR3, respectively, of a VL amino acid sequence of any one or more of SEQ ID NOs:25-39.

[0206] In some embodiments, a polypeptide comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 that substantially preserve one or more functional properties of a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 of a polypeptide selected from any one of AB-1 to AB-15c (e.g., AB-13).

[0207] In some embodiments, a polypeptide comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 comprising only one or more conservative substitutions (e.g., only one or more highly conservative substitutions), relative a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 of a polypeptide selected from any one of AB-1 to AB-15c (e.g., AB- 13).

[0208] In some embodiments, a polypeptide comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 comprising up to 1, 2, or 3 conservative substitutions (e.g., up to 1, 2, or 3 highly conservative substitutions), relative a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 of a polypeptide selected from any one of AB-1 to AB-15c (e.g., AB- 13).

[0209] In some embodiments, a polypeptide disclosed herein comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 having 100% sequence identity to a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3 of a polypeptide selected from any one of AB-1 to AB-15c (e.g., AB-13).

[0210] In some embodiments, a polypeptide comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2 and a LCDR3, of an antibody comprising a VH / VL combination selected from:SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); or SEQ ID NO:20 and SEQ ID NO:39 (AB-15c).

[0211] In some embodiments, a polypeptide comprises: a) a HCDR1 comprising at least one amino acid substitution relative to at least one amino acid sequence set forth in SEQ ID NOs:43-45 (e.g., at least one amino acid sequence set forth in SEQ ID NO:44 or SEQ ID NO:45); b) a HCDR2 comprising at least one amino acid substitution relative to at least one amino acid sequence set forth in SEQ ID NOs:47-53 (e.g., at least one amino acid sequence set forth in SEQ ID NOs:48-53); c) a HCDR3 comprising at least one amino acid substitution relative to at least one amino acid sequence set forth in SEQ ID NOs:55-70 e.g., at least one amino acid sequence set forth in SEQ ID NOs:56-70); d) a LCDR1 comprising at least one amino acid substitution relative to the amino acid sequence set forth in SEQ ID NOs:72-82 e.g., at least one amino acid sequence set forth in SEQ ID NOs:73-82);e) a LCDR2 comprising at least one amino acid substitution relative to at least one amino acid sequence set forth in SEQ ID NO:84 or SEQ ID NO:85 (e.g., SEQ ID NO:85); f) a LCDR3 comprising at least one amino acid substitution relative to at least one amino acid sequence set forth in SEQ ID NOs:87-97 (e.g., at least one amino acid sequence set forth in SEQ ID NOs:88-97); or any combination of the foregoing.Paratopes

[0212] Amino acid residues of a paratope contribute to an antibody’s interaction with an epitope of its target protein. An interaction can be a hydrogen bond, a salt bridge, a van der Waals interaction, an electrostatic interaction, a hydrophobic interaction, pi-interaction effects, an ionic bond, and / or any combination thereof. An interaction can be direct, or indirect, e.g., via a coordinated intermediate molecule, such as an ion or water. The residues of a paratope, in some embodiments, comprise only residues that are part of a defined CDR. In some embodiments, the residues of a paratope further comprise one or more residues that are not part of a defined CDR (e.g., residues within a defined framework region).

[0213] In some embodiments, a paratope is oriented less than about 5.0 angstroms from an epitope on a target antigen when a polypeptide is bound to the target antigen, e.g., less than about: 4.5, 4.0, 3.5, 3.0, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0 or 0.9 angstroms, or about: 0.9-5.0, 0.9-4.8. 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, from the epitope. In some embodiments, less than all of the amino acid residues constituting a paratope (e.g., about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% of the amino acid residues) in the paratope are oriented less than about 5.0 angstroms from an epitope on a target antigen when a polypeptide is bound to the target antigen.

[0214] In some embodiments, a polypeptide comprising a paratope disclosed herein comprises a VH and a VL. In some embodiments, paratope residues are contained within the VH and VL of a polypeptide.

[0215] In some embodiments, a polypeptide comprises a paratope that differs from a paratope of an antibody comprising a VH / VL combination of SEQ ID NO:5 / SEQ ID NO:24.

[0216] In some embodiments, a polypeptide comprises a paratope that differs from a paratope of an antibody comprising a VH / VL combination of SEQ ID NO:5 / SEQ ID NO:24, by substitution (e.g., conservative substitution such as highly conservative substitution) of from 1 to 3 (e.g., 1, 2 or 3) residues.

[0217] In some embodiments, a polypeptide comprises a paratope that is substantially similar (e.g., having at least about 90% sequence identity) to a paratope of an antibody comprising a VH / VL combination of SEQ ID NO:5 / SEQ ID NO:24. In some embodiments, a polypeptide comprises a paratope that is substantially similar (e.g, having at least about 90% sequence identity) to, and substantially preserves one or more functional properties of, a paratope of an antibody comprising a VH / VL combination of SEQ ID NO:5 / SEQ ID NO:24.

[0218] In some embodiments, a polypeptide comprises a paratope that is identical to a paratope of an antibody comprising a VH / VL combination of SEQ ID NO:5 / SEQ ID NO:24

[0219] In some embodiments, a polypeptide comprises a paratope that differs from a paratope of an antibody comprising a VH / VL combination selected from: SEQ ID NO:6 / SEQ ID NO:25 (AB-1), SEQ ID NO:7 / SEQ ID NO:26 (AB-2), SEQ ID NO:8 / SEQ ID NO:27 (AB-3), SEQ ID NO:9 / SEQ ID NO:28 (AB-4), SEQ ID NO: 10 / SEQ ID NO:29 (AB-5), SEQ ID NO: 11 / SEQ ID NO:30 (AB-6), SEQ ID NO: 12 / SEQ ID NO:31 (AB-7), SEQ ID NO: 13 / SEQ ID NO:32 (AB-8), SEQ ID NO: 14 / SEQ ID NO:33 (AB-9), SEQ ID NO: 15 / SEQ ID NO : 34 (AB- 10), SEQ ID NO : 16 / SEQ ID NO : 35 (AB- 11 ), SEQ ID NO : 17 / SEQ ID NO : 36 (AB-12), SEQ ID NO: 18 / SEQ ID NO:37 (AB-13), SEQ ID NO: 19 / SEQ ID NO:38 (AB-14c), or SEQ ID NO:20 / SEQ ID NO:39 (AB-15c).

[0220] See Table 2 for SEQ ID NOs:6-20 and SEQ ID NOs:25-39, and FIGs. 2A-3B for the paratope residues of antibodies comprising VH sequences set forth in SEQ ID NOs:6-20 and VL sequences set forth in SEQ ID Nos:25-39.

[0221] In some embodiments, a polypeptide comprises a paratope that differs from a paratope of an antibody comprising a VH / VL combination selected from: SEQ ID NO:6 / SEQ ID NO:25 (AB-1), SEQ ID NO:7 / SEQ ID NO:26 (AB-2), SEQ ID NO:8 / SEQ ID NO:27 (AB-3), SEQ ID NO:9 / SEQ ID NO:28 (AB-4), SEQ ID NO: 10 / SEQ ID NO:29 (AB-5), SEQ ID NO: 11 / SEQ ID NO:30 (AB-6), SEQ ID NO: 12 / SEQ ID NO:31 (AB-7), SEQ ID NO: 13 / SEQ ID NO:32 (AB-8), SEQ ID NO: 14 / SEQ ID NO:33 (AB-9), SEQ ID NO: 15 / SEQ ID NO : 34 (AB- 10), SEQ ID NO : 16 / SEQ ID NO : 35 (AB- 11 ), SEQ ID NO : 17 / SEQ ID NO : 36 (AB-12), SEQ ID NO: 18 / SEQ ID NO:37 (AB-13), SEQ ID NO: 19 / SEQ ID NO:38 (AB-14c),or SEQ ID NO:20 / SEQ ID NO:39 (AB-15c), by substitution (e.g., conservative substitution such as highly conservative substitution) of from 1 to 3 (e.g., 1, 2 or 3) residues.

[0222] In some embodiments, a polypeptide comprises a paratope that is substantially similar (e.g., having at least about 90% sequence identity) to a paratope of an antibody comprising a VH / VL combination selected from:SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NOV and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); orSEQ ID NOVO and SEQ ID NO:39 (AB-15c); or any combination of the foregoing.

[0223] In some embodiments, a polypeptide comprises a paratope that is substantially similar (e.g., having at least about 90% sequence identity) to, and substantially preserves one or more functional properties of, a paratope of an antibody comprising a VH / VL combination selected from:SEQ ID NOV and SEQ ID NO:25 (AB-1);SEQ ID NOV and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NOV and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); orSEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or any combination of the foregoing.

[0224] In some embodiments, a polypeptide comprises a paratope comprising only one or more conservative substitutions (e.g., only one or more highly conservative substitutions) relative a paratope of an antibody comprising a VH / VL combination selected from:SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NOV and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); orSEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or any combination of the foregoing.

[0225] In some embodiments, a polypeptide comprises a paratope comprising up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 conservative substitutions (e.g., up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 highly conservative substitutions), relative a paratope of an antibody comprising a VH / VL combination selected from:SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID N0:8 and SEQ ID NO:27 (AB-3);SEQ ID N0:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); orSEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or any combination of the foregoing.

[0226] In some embodiments, a polypeptide comprises a paratope that has 100% sequence identity to a paratope of an antibody comprising a VH / VL combination selected from:SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); orSEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or any combination of the foregoing.

[0227] In some embodiments, a polypeptide comprises a paratope that differs from a paratope of an antibody comprising a VH / VL combination of SEQ ID NO: 18 / SEQ ID NO:37 (AB-13).

[0228] In some embodiments, a polypeptide comprises a paratope that differs from a paratope of an antibody comprising a VH / VL combination of SEQ ID NO: 18 / SEQ ID NO:37 (AB- 13), by substitution (e.g., conservative substitution such as highly conservative substitution) of from 1 to 3 (e.g., 1, 2 or 3) residues.

[0229] In some embodiments, a polypeptide comprises a paratope that is substantially similar (e.g., having at least about 90% sequence identity) to a paratope of an antibody comprising a VH / VL combination of SEQ ID NO: 18 / SEQ ID NO:37 (AB-13).

[0230] In some embodiments, a polypeptide comprises a paratope that is substantially similar (e.g., having at least about 90% sequence identity) to, and substantially preserves one or more functional properties of, a paratope of an antibody comprising a VH / VL combination of SEQ ID NO: 18 / SEQ ID NO:37 (AB- 13).

[0231] In some embodiments, a polypeptide comprises a paratope comprising only one or more conservative substitutions (e.g, only one or more highly conservative substitutions) relative a paratope of an antibody comprising a VH / VL of SEQ ID NO: 18 / SEQ ID NO:37 (AB-13).

[0232] In some embodiments, a polypeptide comprises a paratope comprising up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 conservative substitutions (e.g., up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 highly conservative substitutions), relative a paratope of an antibody comprising a VH / VL combination of SEQ ID NO: 18 / SEQ ID NO:37 (AB-13).

[0233] In some embodiments, a polypeptide comprises a paratope that has 100% sequence identity to a paratope of an antibody comprising a VH / VL combination of SEQ ID NO: 18 / SEQ ID NO:37 (AB-13).

[0234] In some embodiments, a polypeptide comprises a paratope comprising amino acid residues corresponding to each of X2 (position 30), A33, T35, S50, 151, S52, G53, S54, X4 (position 57), Y59, D99, X7 (position 100), X9 (position 102), X10 (position 103), Xu (position 104), X12 (position 105), X13 (position 106), Pl 07, and X14 (position 108) of SEQ ID NO:4, and Y31, S32, Y37, X21 (position 96), L97, X22 (position 98), X23 (position T99), and Y101 of SEQ ID NO:23, or a subset thereof.Consensus Sequences

[0235] In some embodiments, a polypeptide comprises a VH comprising the amino acid sequence of SEQ ID NO:4, wherein:Xi is not T;X2 is not R;X3 is not G;X4 is not N;X5 is not T;Xe is not K;X7 is not R;Xs is not L;Xg is not S;Xio is not I;Xu is not T;X12 is not I;X13 is not R;X14 is not R;X15 is not Y; orXi6 is not V; or any combination of the foregoing.

[0236] The sequence identified as SEQ ID NO:4 is shown in Table 2, which is a consensus VH sequence for SEQ ID Nos:5-20 herein.

[0237] In some embodiments:Xi is T or N;X2 is R, S or K;X3 is G, S, D or T;X4 is N, T or K;X5 is T, K or I;Xe is K or R;X7 is R or A;Xs is L, W or Y;Xg is S or T;X10 is I, A, K, T, S or V;Xu is T, A, K, S or R;X12 is I, V, A or T;X13 is R or K;Xi4 is R, Y, E, I, N, S, V or K;X15 is Y, V or R; orXi6 is V or I, or any combination of the foregoing.

[0238] In some embodiments:Xi is N;X2 is S or K;X3is S, D or T;X4is T or K;X5is K or I;Xe is R;X7 is A;Xs is W or Y;X9is T;X10 is A, K, T, S or V;Xn is A, K, S or R;X12 is V, A or T;X13 is K;X14 is Y, E, I, N, S, V or K;X15 is V or R; orXi6 is I; or any combination of the foregoing.

[0239] In some embodiments, Xi is not T. In some embodiments, Xi is T or N. In some embodiments, Xi is T. In some embodiments, Xi is N.

[0240] In some embodiments, X2 is not R. In some embodiments, X2 is R, S or K. In some embodiments, X2 is S or K. In some embodiments, X2 is R. In some embodiments, X2 is S. In some embodiments, X2 is K.

[0241] In some embodiments, X3 is not G. In some embodiments, X3 is G, S, D or T. In some embodiments, X3 is S, D or T. In some embodiments, X3 is G. In some embodiments, X3 is S. In some embodiments, X3 is D. In some embodiments, X3 is T.

[0242] In some embodiments, X4 is not N. In some embodiments, X4 is N, T or K. In some embodiments, X4 is T or K. In some embodiments, X4 is N. In some embodiments, X4 is T. In some embodiments, X4 is K.

[0243] In some embodiments, X5 is not T. In some embodiments, X5 is T, K or I. In some embodiments, X5 is K or I. In some embodiments, X5 is T. In some embodiments, X5 is K. In some embodiments, X5 is I.

[0244] In some embodiments, Xe is not K. In some embodiments, Xe is K or R. In some embodiments, Xe is K. In some embodiments, Xe is R.

[0245] In some embodiments, X7 is not R. In some embodiments, X7 is R or A. In some embodiments, X7 is R. In some embodiments, X7 is A.

[0246] In some embodiments, Xs is not L. In some embodiments, Xs is L, W or Y. In some embodiments, Xs is W or Y. In some embodiments, Xs is L. In some embodiments, Xs is W. In some embodiments, Xs is Y.

[0247] In some embodiments, X9 is not S. In some embodiments, X9 is S or T. In some embodiments, X9 is S. In some embodiments, X9 is T.

[0248] In some embodiments, X10 is not I. In some embodiments, X10 is I, A, K, T, S or V. In some embodiments, X10 is A, K, T, S or V. In some embodiments, X10 is I. In some embodiments, X10 is A. In some embodiments, X10 is K. In some embodiments, X10 is T. In some embodiments, X10 is S. In some embodiments, X10 is V.

[0249] In some embodiments, X11 is not T. In some embodiments, X11 is T, A, K, S or R. In some embodiments, X11 is A, K, S or R. In some embodiments, X11 is T. In some embodiments, X11 is A. In some embodiments, X11 is K. In some embodiments, X11 is S. In some embodiments, X11 is R.

[0250] In some embodiments, X12 is not I. In some embodiments, X12 is I, V, A or T. In some embodiments, X12 is V, A or T. In some embodiments, X12 is I. In some embodiments, X12 is V. In some embodiments, X12 is A. In some embodiments, X12 is T.

[0251] In some embodiments, X13 is not R. In some embodiments, X13 is R or K. In some embodiments, X13 is R. In some embodiments, X13 is K.

[0252] In some embodiments, X14 is not R. In some embodiments, X14 is R, Y, E, I, N, S, V or K. In some embodiments, X14 is Y, E, I, N, S, V or K. In some embodiments, X14 is R. In some embodiments, X14 is Y. In some embodiments, X14 is E. In some embodiments, X14 is I. In some embodiments, X14 is N. In some embodiments, X14 is S. In some embodiments, X14 is V. In some embodiments, X14 is K.

[0253] In some embodiments, X15 is not Y. In some embodiments, X15 is Y, V or R. In some embodiments, X15 is V or R. In some embodiments, X15 is Y. In some embodiments, X15 is V. In some embodiments, X15 is R.

[0254] In some embodiments, Xi6 is not V. In some embodiments, Xi6 is V or I. In some embodiments, Xi6 is V. In some embodiments, Xi6 is I.

[0255] In some embodiments, a polypeptide comprises a VL comprising the amino acid sequence of SEQ ID NO:23, wherein:X17 is not L;Xis is not I;X19 is not Y;X20 is not G;X21 is not A;X22 is not Q; orX23 is not T; or any combination of the foregoing.

[0256] The sequence identified as SEQ ID NO:23 is shown in Table 2, which is a consensus VL sequence for SEQ ID Nos:24-39 herein.

[0257] In some embodiments:X17 is L or D;Xis is I, Y, S, T, E, L, V, Q, H or R;X19 is Y or K;X20 is G or A;X21 is A, S, T or D;X22 is Q, S, K, T, F, H or Y; orX23 is T or H; or any combination of the foregoing.

[0258] In some embodiments:X17 is D;Xis is Y, S, T, E, L, V, Q, H or R;X19 is K;X20 is A;X21 is S, T or D;X22 is S, K, T, F, H or Y; orX23 is H; or any combination of the foregoing.

[0259] In some embodiments, X17 is not L. In some embodiments, X17 is L or D. In some embodiments, X17 is L. In some embodiments, X17 is D.

[0260] In some embodiments, Xis is not I. In some embodiments, Xis is I, Y, S, T, E, L, V, Q, H or R. In some embodiments, Xis is Y, S, T, E, L, V, Q, H or R. In some embodiments, Xis is Y. In some embodiments, Xis is S. In some embodiments, Xis is T. In some embodiments, Xis is E. In some embodiments, Xis is L. In some embodiments, Xis is V. In some embodiments, Xis is Q. In some embodiments, Xis is H. In some embodiments, Xis is R.

[0261] In some embodiments, X19 is not Y. In some embodiments, X19 is Y or K. In some embodiments, X19 is Y. In some embodiments, X19 is K.

[0262] In some embodiments, X20 is not G. In some embodiments, X20 is G or A. In some embodiments, X20 is G. In some embodiments, X20 is A.

[0263] In some embodiments, X21 is not A. In some embodiments, X21 is A, S, T or D. In some embodiments, X21 is S, T or D. In some embodiments, X21 is S. In some embodiments, X21 is T. In some embodiments, X21 is D.

[0264] In some embodiments, X22 is not Q. In some embodiments, X22 is Q, S, K, T, F, H or Y. In some embodiments, X22 is S, K, T, F, H or Y. In some embodiments, X22 is S. In some embodiments, X22 is K. In some embodiments, X22 is T. In some embodiments, X22 is F. In some embodiments, X22 is H. In some embodiments, X22 is Y.

[0265] In some embodiments, a) Xi is T, X2 is S, X3 is G, X4is N, X5is T, X6is K, X7 is A, X8is L, X9is T, X10 is I, Xu is A, X12 is I, X13 is R, Xi4is Y, X15 is Y, Xi6 is V, X17 is D, Xis is Y, X19 is Y, X20 is A, X21 is S, X22 is S, X23 is T, or a combination thereof (AB-1); b) Xi is T, X2 is R, X3 is S, X4is N, X5is T, X6is R, X7 is R, X8is L, X9is T, X10 is A, Xu is A, X12 is V, X13 is R, Xi4is Y, X15 is Y, Xi6 is V, X17 is D, Xis is S, X19 is Y, X20 is A, X21 is S, X22 is Q, X23 is T, or a combination thereof (AB-2); c) Xi is T, X2 is R, X3is G, X4is N, X5is T, X6is K, X7 is R, X8is L, X9is S, X10 is K, Xu is T, X12 is I, X13 is R, Xi4is Y, X15 is Y, Xi6is V, X17 is L, Xisis T, X19 is Y, X20 is G, X21 is S, X22 is S, X23 is T, or a combination thereof (AB-3); d) Xi is T, X2 is S, X3is G, X4is N, X5is T, X6is R, X7is R, Xs is L, X9 is T, X10 is I, Xu is T, X12 is I, X13 is R, Xi4is R, X15 is Y, Xi6 is V, X17 is L, Xis is T, X19 is Y, X20 is A, X21 is A, X22 is S, X23 is T, or a combination thereof (AB-4); e) Xi is T, X2 is S, X3 is S, X4is N, X5is T, X6is K, X7 is R, Xs is L, X9is T, X10 is I, Xu is A, X12 is I, X13 is R, Xi4is Y, X15 is Y, Xi6 is V, X17 is D, Xis is I, X19 is Y, X20 is A, X21 is A, X22 is Q, X23 is T, or a combination thereof (AB- 5); f) X1 is T, X2 is R, X3is S, X4is N, X5is T, X6is K, X7 is R, X8is L, X9is T, X10 is T, Xu is K, X12 is V, X13 is R, Xi4is E, X15 is Y, Xi6 is V, X17 is L, Xis is I, X19 is Y, X20 is A, X21 is S, X22 is Q, X23 is T, or a combination thereof (AB-6); g) Xi is T, X2 is R, X3is S, X4is T, X5is T, X6is K, X7 is R, X8is L, X9is T, X10 is A, Xu is S, X12 is V, X13 is R, Xi4is I, X15 is V, Xi6 is V, X17 is L, Xis is E, X19 is Y, X20 is A, X21 is T, X22 is K, X23 is T, or a combination thereof (AB- 7); h) Xi is T, X2 is R, X3 is S, X4is N, X5is T, X6is K, X7 is R, X8is L, X9is T, X10 is A, Xu is S, X12 is V, X13 is R, Xi4is N, X15 is Y, Xi6is V, X17 is L, Xis is E, X19 is Y, X20 is A, X21 is S, X22 is T, X23 is T, or a combination thereof (AB-8); i) Xi is T, X2 is R, X3 is S, X4is N, X5is T, X6is K, X7 is R, Xs is L, X9is T, X10 is T, Xu is A, X12 is V, X13 is R, Xi4is S, X15 is Y, Xi6 is V, X17 is L, Xis is L, X19 is Y, X20 is A, X21 is S, X22 is Q, X23 is T, or a combination thereof (AB-9); j) Xi is T, X2 is R, X3is D, X4is K, X5is T, X6is R, X7 is R, Xs is L, X9is S, X10 is S, Xu is S, X12 is A, X13 is R, Xi4is V, X15 is V, Xi6is V, X17 is L, Xis is V, X19 is K, X20 is A, X21 is D, X22 is F, X23 is H, or a combination thereof (AB- 10); k) Xi is T, X2 is R, X3 is T, X4is N, X5is T, X6is K, X7 is R, Xs is L, X9is S, X10 is V, Xu is S, X12 is T, X13 is R, Xi4is K, X15 is Y, Xi6is V, X17 is D, Xisis Y, X19 is Y, X20 is A, X21 is S, X22 is H, X23 is T, or a combination thereof (AB-11); l) Xi is T, X2 is R, X3is S, X4is N, X5is K, X6is K, X7is R, X8is L, X9 is T, X10 is S, Xu is R, X12 is V, X13 is R, Xi4is Y, X15 is Y, Xi6is V, X17 is L, Xis is Q, X19 is Y, X20 is A, X21 is S, X22 is F, X23 is T, or a combination thereof (AB- 12); m) Xi is T, X2 is R, X3 is S, X4is N, X5is T, X6is K, X7 is R, X8is W, X9is T, X10 is S, Xu is S, X12 is V, X13 is R, Xi4is V, X15 is R, Xi6 is I, X17 is L, Xis is H, X19 is Y, X20 is A, X21 is S, X22 is Y, X23 is T, or a combination thereof (AB-13); n) Xi is T, X2 is R, X3 is S, X4is N, X5is I, X6is K, X7 is R, X8is Y, X9is T, X10 is K, Xu is S, X12 is V, X13 is R, Xi4is V, X15 is R, Xi6 is I, X17 is L, Xis is H, X19 is Y, X20 is A, X21 is S, X22 is K, X23 is T, or a combination thereof (AB- 14c); o) Xi is N, X2 is K, X3is S, X4is N, X5is T, X6is K, X7 is R, X8is W, X9is T, X10 is S, Xu is S, X12 is V, X13 is K, Xi4is R, X15 is R, Xi6 is I, X17 is L, Xis is R, X19 is Y, X20 is A, X21 is S, X22 is Y, X23 is T, or a combination thereof (AB-15c).Constant Domains

[0266] In some embodiments, a polypeptide comprises: a) an antibody heavy chain constant domain sequence; b) an antibody light chain constant domain sequence; or both a) and b).

[0267] In some embodiments, a polypeptide comprises an antibody heavy chain constant domain sequence. In some embodiments, an antibody heavy chain constant domain is selected from the group consisting of an IgA constant domain, an IgD constant domain, an IgE constant domain, an IgG constant domain and an IgM constant domain.

[0268] In some embodiments, an IgG constant domain is an IgGl constant domain, an IgG2 constant domain, an IgG3 constant domain or an IgG4 constant domain. In some embodiments, the IgG2 constant domain is an IgG2a, an IgG2b constant domain or an IgG2c constant domain. In some embodiments, an IgA constant domain is an IgAl constant domainor an IgA2 constant domain. In some embodiments, the antibody heavy chain constant domain is an IgGl constant domain (e.g., IGHV1-5 or IGHV5-51).

[0269] In some embodiments, the name of a polypeptide comprising a human IgGl (hlgGl) domain is followed by “d”. In some embodiments, the name of a polypeptide comprising a human IgG4 (hIgG4) domain is followed by “c”. In some embodiments, the name of a polypeptide comprising a human IgG4 LS variant (hIgG4 LS) domain is followed by “b”. In some embodiments, the name of a polypeptide comprising a human IgG4 YTE variant (hIgG4 YTE) domain is followed by “a”.

[0270] In some embodiments, an antibody heavy chain constant domain sequence has at least about 60% sequence identity to the amino acid sequence of any one of SEQ ID NOs:98- 101. For example, an antibody heavy chain constant domain sequence can have at least about: 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of any one of SEQ ID NOs:98-101. In some embodiments, the antibody heavy chain constant domain sequence has at least about 70% or at least about 80% sequence identity to the amino acid sequence of any one of SEQ ID NOs:98-101.

[0271] In some embodiments, an antibody heavy chain constant domain sequence comprises at least one amino acid substitution relative to the amino acid sequence of any one of SEQ ID NOs:98-101. 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, an antibody heavy chain constant domain sequence comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of any one of SEQ ID NOs:98-101.

[0272] In some embodiments, a polypeptide comprises an Fc polypeptide, or Fc domain (e.g., an IgG4 domain). In certain embodiments, the Fc domain comprises a mutation that decreases (e.g., inhibits, ablates) an effector function of the Fc domain. See, e.g., Dumet et al., Insights into the IgG heavy chain engineering patent landscape as applied to IgG4 antibody development, MAbs. 11(8): 1341-50 (2019) (particularly Tables 1 and 2 therein) and W002060919, the contents of which are incorporated by reference herein in their entirety. In particular embodiments, the Fc domain comprises LS (M428L / N434S by Kabat numbering) or YTE (M252Y / S254T / T256E by Kabat numbering).

[0273] In some embodiments, a polypeptide comprises an antibody light chain constant domain sequence.

[0274] In some embodiments, an antibody light chain constant domain is selected from the group consisting of a K constant domain and a X constant domain.

[0275] In some embodiments, an antibody light chain constant domain sequence has at least about 60% sequence identity to the amino acid sequence of SEQ ID NO: 102 or SEQ ID NO: 103. For example, an antibody light chain constant domain sequence can have at least about: 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 102 or SEQ ID NO: 103. In some embodiments, an antibody light chain constant domain sequence has at least about 70% or at least about 80% sequence identity to SEQ ID NO: 102 or SEQ ID NO: 103.

[0276] In some embodiments, an antibody light chain constant domain sequence comprises at least one amino acid substitution relative to the amino acid sequence of SEQ ID NO: 102 or SEQ ID NO: 103. 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, an antibody light chain constant domain sequence comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of SEQ ID NO: 102 or SEQ ID NO: 103. In some embodiments, the at least one amino acid substitution is a conservative substitution. In some embodiments, the at least one amino acid substitution is a highly conservative substitution.

[0277] In some embodiments, a polypeptide comprises: a) an antibody heavy chain constant domain sequence; and b) an antibody light chain constant domain sequence.

[0278] In some embodiments, an antibody heavy chain constant domain is an IgG4 constant domain (e.g., SEQ ID NO:98), and an antibody light chain constant domain is a K constant domain (e.g., SEQ ID NO: 102).Antibodies and Antigen Binding Fragments

[0279] In some embodiments, a 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.

[0280] In some embodiments, a polypeptide is an antibody. As used herein, the term “antibody” refers to an immunoglobulin molecule capable of specific binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site, located in the variable domain of the immunoglobulin molecule. As used herein, the term “antibody” refers to a full-length antibody.

[0281] In some embodiments, a polypeptide is an antibody comprising two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds or multimers thereof (for example, IgM). Each heavy chain comprises a VH and a heavy chain constant domain (comprising domains CHI, hinge CH2 and CH3). Each light chain comprises a VL and a light chain constant domain (CL). VH and VL regions may be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed within framework regions (FRs). VH and VL each comprises three CDRs and four FRs, arranged from the amino-terminus to the carboxy -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. An antibody can be of any species, such as a murine antibody, a human antibody, or a humanized antibody.

[0282] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 105-160. In some embodiments, a polypeptide comprises a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-176.

[0283] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 105-117. In some embodiments, a polypeptide comprises a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-174. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 105-117, and a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-174.

[0284] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 131-145. In some embodiments, a polypeptide comprises a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-176. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 131-145, and a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-176.

[0285] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 148-160. In some embodiments, a polypeptide comprises a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-174. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 148-160, and a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-174.

[0286] In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 118-130. In some embodiments, a polypeptide comprises a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-174. In some embodiments, a polypeptide comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 118-130, and a light chain amino acid sequence set forth in any one of SEQ ID NOs: 162-174.

[0287] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 118; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 162 (AB- 1b).

[0288] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 119; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 163 (AB- 2b).

[0289] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 120; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 164 (AB- 3b).

[0290] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 121; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 165 (AB- 4b).

[0291] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 122; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 166 (AB- 4B).

[0292] In some embodiments, a polypeptide comprises:a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 123; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 167 (AB- 66).

[0293] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 124; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 168 (AB- 7b).

[0294] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 125; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 169 (AB- 8b).

[0295] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 126; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 170 (AB- 9b).

[0296] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 127; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 171 (AB- 10b).

[0297] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 128; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 172 (AB- 11b).

[0298] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 129; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 173 (AB- 12b).

[0299] In some embodiments, a polypeptide comprises: a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 130; and b) a light chain comprising the amino acid sequence of SEQ ID NO: 174 (AB- 13b).

[0300] In some embodiments, a polypeptide is a single-domain antibody or an antigenbinding fragment thereof. As used herein, the term “single-domain antibody (sdAb)” or“nanobody” refers to an immunoglobulin molecule consisting of a single monomeric variable antibody domain and capable of specific binding to a target. A single-domain antibody can be of any species, such as a murine antibody, a human antibody or a humanized single-domain antibody.

[0301] In some embodiments, a VH domain and a VL domain may be linked together via a linker (e.g., a synthetic linker) to form various types of single-chain antibody designs in which the VH / VL domains pair intramolecularly, or intermolecularly in those cases when the VH and VL domains are expressed by separate chains, to form a monovalent antigen binding site.

[0302] In some embodiments, a polypeptide is a heavy-chain antibody comprising two or more heavy chains, but lacking light chain, or an antigen-binding fragment thereof. Nonlimiting examples of heavy chain antibodies include camelid Vhh (also referred to as VHH or VHH) antibodies. Camelid antibodies are antibodies from the Camelidae family of mammals that include llamas, camels, and alpacas.

[0303] In some embodiments, a polypeptide is an antibody mimetic. The term “antibody mimetic” refers to polypeptides capable of mimicking an antibody’s ability to bind an antigen, but structurally differ from native antibody structures. Non-limiting examples of antibody mimetics include Adnectins, Affibodies, Affilins, Affimers, Affitins, Alphabodies, Anticalins, Avimers, DARPins, Fynomers, Kunitz domain peptides, monobodies, nanobodies, nanoCLAMPs, and Versabodies.

[0304] In some embodiments, a polypeptide is an antigen-binding fragment of an antibody. The term “antigen-binding fragment” refers to a portion of an immunoglobulin molecule (e.g., antibody) that retains the antigen binding properties of the full-length antibody. Non-limiting examples of antigen-binding fragments include a VH region, a VL region, an Fab fragment, an F(ab’)2 fragment, an Fd fragment, an Fv fragment, and a domain antibody (dAb) consisting of one VH domain or one VL domain, etc. VH and VL domains may be linked together via a synthetic linker to form various types of single-chain antibody designs in which the VH / VL domains pair intramolecularly, or intermolecularly in those cases when the VH and VL domains are expressed by separate chains, to form a monovalent antigen binding site, such as single chain Fv (scFv) or diabody. In some embodiments, a polypeptide disclosed herein is an antigen binding fragment selected from Fab, F(ab’)2, Fab’, scFv, or Fv. In some embodiments, a polypeptide is a scFv.

[0305] In some embodiments, a polypeptide is an isolated polypeptide.

[0306] In some embodiments, a polypeptide (e.g., an isolated polypeptide) is recombinantly produced. In some embodiments, a polypeptide (e.g., an isolated polypeptide) is synthetically produced.

[0307] In some embodiments, a polypeptide is linked to a second polypeptide. The term “linked” means attached, via a covalent or noncovalent interaction. Conjugation can employ a suitable linking agent. Non-limiting examples include peptide linkers, compound linkers, and chemical cross-linking agents. In some embodiments, the linker is a disulfide bond.

[0308] In some embodiments, a polypeptide is conjugated to a heterologous moiety. The term “conjugated” refers to attached, via a covalent or noncovalent interaction. Conjugation can employ any one or more of suitable linking agents. Non-limiting examples include peptide linkers, compound linkers, and chemical cross-linking agents.

[0309] In some embodiments, a heterologous moiety comprises a therapeutic agent, a diagnostic agent, or both. In some embodiments, a heterologous moiety is selected from polyethylene glycol (PEG), hexadecanoic acid, hydrogels, nanoparticles, multimerization domains and carrier peptides.

[0310] In some embodiments, a nanoparticle is a lipid nanoparticle. In some embodiments, a nanoparticle is a polymer nanoparticle. In some embodiments, a polymer is an amphiphilic polymer. In some embodiments, a polymer is a hydrophobic or hydrophilic polymer. Non-limiting examples of polymers include poly(lactic acid)-poly(ethylene glycol), poly(lactic-co-glycolic acid)-poly(ethylene glycol), poly(lactic-co-glycolic) acid (PLGA), poly(lactic-co-glycolic acid)-d-a-tocopheryl polyethylene glycol succinate, poly(lactic-co- glycolic acid)-ethylene oxide fumarate, poly(glycolic acid)-poly(ethylene glycol), polycaprolactone-poly(ethylene glycol), or any salts thereof. In some embodiments, a polymer nanoparticle comprises poly(lactic-co-glycolic) acid (PLGA).

[0311] In some embodiments, a carrier polypeptide is albumin or an Fc polypeptide.

[0312] In some embodiments, a polypeptide binds an IL-4Ra with a binding constant(KD) of about 1 pM or less. As used herein the term “KD,” also referred to as “binding constant,” “equilibrium dissociation constant” or “affinity constant,” is a measure of the extent of a reversible association between two molecular species (e.g., antibody and target protein) and includes both the actual binding affinity as well as the apparent binding affinity. Binding affinity can be determined using methods known in the art including, for example, by measurement of surface plasmon resonance, e.g., using a Biolayer interferometry (Octet, ForteBio) or a surface plasmon resonance (Biacore) system and assay. A reference thatcompares various surface technologies for measuring binding affinity and kinetics is Yang, D., Singh, A., Wu, H., & Kroe-Barrett, R., Comparison of biosensor platforms in the evaluation of high affinity antibody-antigen binding kinetics, Analytical Biochemistry 508: 78-96 (2016), the contents of which are incorporated by reference herein in their entirety.

[0313] In some embodiments, a polypeptide binds an IL-4Ra with a binding constant (KD) of about 0.05 to 0.5 nM or less or about 0.02 to 0.04 nM or less.

[0314] In some embodiments, a polypeptide binds an IL-4Ra with an association constant (ka) of about 100 x 105M^s'1or less, about 7 x 105to 9 x 105M^s'1or less, or about 7.7 x 105to 8.9 x 105M^s'1or less.

[0315] In some embodiments, a polypeptide dissociates from an IL-4Ra with a dissociation constant (kd) of about about 100 x 10'5s'1or less, 2 x 10'5to 3 x 10'5s'1or less, or about 2.1 x 10'5to 2.6 x 10'5s'1or less.

[0316] In some embodiments, a polypeptide binds an IL-4Ra with an ECso of about 1 pM or less, about 0.02 to 1.5 nM or less, or about 0.04 to 1.3 nM or less.

[0317] In some embodiments, a polypeptide blocks IL-4 Type II signaling with an ICso of about 1 pM or less, about 1.1 to 7.3 nM or less, about 4.4 to 7.3 nM or less, or about 1.1 to 1.8 nM or less.

[0318] In some embodiments, a polypeptide blocks IL- 13 Type II signaling with an ICso of about 1 pM or less or about 3 to 3.3 nM or less.

[0319] In some embodiments, a polypeptide inhibits CD23 expression in B cells with an ICso of about 1 pM or less, about 8 to 50nM or less, about 30 to 50 nM or less, about 30 to 34 nM or less, or about 8 to 8.8 nM or less.

[0320] In some embodiments, a polypeptide competes with the Reference Antibody for binding to an interleukin-4 receptor alpha (IL-4Ra). Techniques and assays for assessing competition between antibodies are known in the art.

[0321] In some embodiments, a polypeptide has a weaker self-association than the Reference Antibody, for example, as determined by an affinity-capture self-interaction nanoparticle spectroscopy (AC-SINS) value. The AC-SINS value is the change in maximum absorbance wavelength in the coated-nanoparticle absorption spectra compared to the spectra of the nanoparticle alone. Thus, the greater the change in maximum absorbance wavelength, the more self-interaction of the antibody coated on the nanoparticle. Self-association is an unwanted property that correlates with poor viscosity and poor PK properties. Techniques and assays for assessing self-association of proteins are known in the art. See, e.g., Patro &Przybycien, Biotechnol Bioeng. 52(2): 193-203 (1996), the contents of which are incorporated herein in their entirety. In some embodiments, a polypeptide has a weaker self-association than the Reference Antibody.

[0322] In some embodiments, a polypeptide has an AC-SINS value of no more than about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 18, 20, 21, 22, 23, 24 or 25. In some embodiments, a polypeptide has an AC-SINS value of no more than about 14. In some embodiments, a polypeptide has an AC-SINS value of no more than about 8. In some embodiments, a polypeptide has an AC-SINS value of about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 18, 20, 21, 22, 23, 24 or 25. In some embodiments, a polypeptide has an AC-SINS value of about 0-25, e.g., 0-20, 0-15, 0-10, 0-8, 0-5, 2-20, 2-15, 2-10, 2-8, 2- 5, 5-20, 5-15, 5-10, 5-8, 7-8 or 13-15. In some embodiments, a polypeptide has an AC-SINS value of about 13-14, 13-15, 7-9 or 7-8. In some embodiments, a polypeptide has an AC- SINS value of about: 8 or 14.

[0323] In some embodiments, a polypeptide has an improved developability (e.g., reduced AC-SINS) relative to the Reference Antibody. In some embodiments, the selfassociation of a polypeptide is at least about 10% lower than that of the Reference Antibody, for example, by at least about: 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% lower than that of the Reference Antibody. In some embodiments, the self-association of a polypeptide is at least about 30% lower than that of the Reference Antibody.

[0324] In some embodiments, the self-association of a polypeptide is less than about 90% of that 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 that of the Reference Antibody.

[0325] In some embodiments, the self-association of a polypeptide is about 1-90% relative to that of 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 that of the Reference Antibody.

[0326] In some embodiments, the reduction in self-association relative to the Reference Antibody is at least about 10%, for example, by at least about: 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.Fusion Proteins

[0327] In some embodiments, the disclosure provides a fusion protein comprising one or more of polypeptides described herein.

[0328] The term “fusion protein” refers to a synthetic, semi-synthetic or recombinant single protein molecule. A fusion protein can comprise all or a portion of two or more different proteins and / or polypeptides that are attached by covalent bonds (e.g., peptide bonds). For example, a fusion protein can comprise a full-length polypeptide disclosed herein (e.g., a whole antibody), or a fragment thereof (e.g., an antigen-binding fragment of an antibody). The heterologous partner can be a full-length protein or a fragment thereof (e.g., a truncated protein).

[0329] Fusion proteins can be produced recombinantly or synthetically, using routine methods and reagents that are well known in the art. For example, a fusion protein disclosed herein can be produced recombinantly in a suitable host cell (e.g., bacteria) according to methods known in the art. See, e.g., Current Protocols in Molecular Biology, Second Edition, Ausubel et al. eds., John Wiley & Sons, 1992; and Molecular Cloning: a Laboratory Manual, 2nd edition, Sambrook et al., 1989, Cold Spring Harbor Laboratory Press. For example, a nucleic acid molecule comprising a nucleotide sequence encoding a fusion protein described herein can be introduced and expressed in suitable host cell (e.g., E. coif), and the expressed fusion protein can be isolated / purified from the host cell (e.g., in inclusion bodies) using routine methods and readily available reagents. For example, DNA fragments coding for different protein sequences (e.g., a light-responsive domain, a heterologous peptide component) can be ligated together in-frame in accordance with conventional techniques. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. In some embodiments, PCR amplification of nucleic acid fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive nucleic acid fragments that can subsequently be annealed and re-amplified to generate a chimeric nucleic acid sequence (see Ausubel et al., Current Protocols in Molecular Biology, 1992).Nucleic Acids, Vectors., Host Cells

[0330] In some embodiments, the disclosure provides one or more polynucleotides (e.g., DNA, RNA, or analogs of either, e.g., optionally including one or more modified nucleotides; the polynucleotide may be linear or circular, e.g., linear or circular RNA) encoding any oneof polypeptides or fusion proteins described herein. In some embodiments, a polypeptide or fusion protein disclosed herein is encoded by a single polynucleotide. In some embodiments, a polypeptide or fusion protein disclosed herein is encoded by multiple polynucleotides.

[0331] In some embodiments, the polynucleotide comprises a nucleotide sequence that is codon-optimized for a chosen host cell.

[0332] In some embodiments, the disclosure provides a vector (e.g., an expression vector, including a viral-delivery vector) comprising any one or more of the polynucleotides described herein.

[0333] The term “expression vector” refers to a replicable nucleic acid from which one or more proteins can be expressed when the expression vector is transformed into a suitable expression host cell.

[0334] In some embodiments, the vector (e.g., expression vector) comprises an expression control polynucleotide sequence operably linked to the polynucleotide, a polynucleotide sequence encoding a selectable marker, or both. 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 region of DNA to which RNA polymerase binds and initiates the transcription of a gene. The term “operably linked” means that the nucleic acid is positioned in the recombinant polynucleotide, e.g., vector, in such a way that enables expression of the nucleic acid under control of the element (e.g., promoter) to which it is linked. The term “selectable marker element” is an element that confers a trait suitable for artificial selection. Selectable marker elements can be negative or positive selection markers.

[0335] In some embodiments, the disclosure provides an expression host cell comprising any one or more of the polynucleotides or expression vectors described herein.

[0336] The term “expression host cell” refers to a cell useful for receiving, maintaining, reproducing and / or amplifying a vector.

[0337] Non-limiting examples of expression host cells include mammalian cells such as hybridoma cells, Chinese hamster ovary (CHO) cells, COS cells, human embryonic kidney (HEK), yeast cells such as Pichia pastoris cells, or bacterial cells such as E. coh. including DH5a, etc.Compositions

[0338] In some embodiments, the disclosure provides a composition comprising any one of polypeptides or fusion proteins described herein. In some embodiments, the composition is a pharmaceutical composition.

[0339] In some embodiments, the composition (e.g., pharmaceutical composition) comprises pharmaceutically acceptable carriers, excipients, stabilizers, diluents or tonifiers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Suitable pharmaceutically acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed. Non-limiting examples of pharmaceutically acceptable carriers, excipients, stabilizers, diluents or tonifiers include buffers (e.g., phosphate, citrate, histidine), antioxidants (e.g., ascorbic acid or methionine), preservatives, proteins (e.g., serum albumin, gelatin or immunoglobulins); hydrophilic polymers, amino acids, carbohydrates (e.g., monosaccharides, disaccharides, glucose, mannose or dextrins); chelating agents (e.g., EDTA), sugars (e.g., sucrose, mannitol, trehalose or sorbitol), saltforming counter-ions (e.g., sodium), metal complexes (e.g., Zn-protein complexes); non-ionic surfactants e.g., Tween), PLURONICS™ and polyethylene glycol (PEG).

[0340] In some embodiments, the composition (e.g, pharmaceutical composition) disclosed herein is formulated for a suitable administration schedule and route. Non-limiting examples of administration routes include oral, rectal, mucosal, intravenous, intramuscular, subcutaneous and topical, etc. In some embodiments, the composition (e.g, pharmaceutical composition) disclosed herein is stored in the form of an aqueous solution or a dried formulation (e.g., lyophilized).

[0341] In some embodiments, the composition is formulated to be administered by infusion (e.g., intravenous infusion).

[0342] In some embodiments, the composition is formulated to be administered with a second therapeutic agent as a combination therapy. In some embodiments, the second therapeutic agent is any one of polypeptides described herein. In some embodiments, the second therapeutic agent comprises an agent indicated for treatment of an inflammatory condition, non-limiting examples of which include: atopic dermatitis, asthma, eczema (atopic dermatitis), food allergy, prurigo nodularis, chronic rhinosinusitis with nasal polyps, keloids, eosinophilic esophagitis, prostate cancer, chronic urticaria, bullous pemphigoid, localized scleroderma, alopecia areata, ulcerative colitis, aspirin-exacerbated respiratory disease, metastatic non-small cell lung cancer, Netherton syndrome, and combinations thereof.

[0343] In some embodiments, non-limiting examples of the second therapeutic agent include: an additional anti-IL-4Ra antibody or antigen-binding fragment thereof, an inhaled corticosteroid, a leukotriene modifier, a combination corticosteroid / long-acting beta agonist inhaler, a bronchodilator, a short-acting beta agonist, an anticholinergic agent, an oral or intravenous corticosteroid, a calcineurin inhibitor, an anti-inflammatory agent, an antirheumatic agent, an immunosuppressant, a biologic agent, an antihistamine, an epinephrine, an oral immunotherapy, a topical corticosteroid, a topical anesthetic, an immunomodulatory agent, a nasal corticosteroid, an antifungal agent, an anti-androgen agent, a luteinizing hormone-releasing hormone (LHRH) agonist, an gonadotropin-releasing hormone (GnRH) antagonist, a chemotherapy, a targeted drug therapy, a steroid-sparing agent, and combinations thereof.Methods of Use

[0344] In some embodiments, the disclosure provides methods of treating a subject in need thereof, comprising administering an effective amount of the composition disclosed herein. In some embodiments, the composition comprises a pharmaceutically acceptable carrier and, wherein as an active ingredient, any one of polypeptides or fusion proteins described herein.

[0345] In some embodiments, the amount of pathogenic IL-4 / IL-13 signaling in the subject is reduced by at least about 10%, e.g., by at least about: 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0346] In some embodiments, the likelihood of pathogenic IL-4 / IL-13 signaling in the subject in the presence of a polypeptide is about 1-90% relative to the likelihood in the absence of the polypeptide, 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%.

[0347] The term “subject” and “patient” are used herein interchangeably to refer to an animal (e.g., a mammal, such as a human) who is to be treated according to a method disclosed herein. A subject to be treated according to methods described herein may be one who has been diagnosed with a particular condition (e.g., in inflammatory condition), or one at risk of developing such conditions or symptoms of such conditions. Diagnosis may be performed by any method or technique known in the art. One skilled in the art willunderstand that a subject to be treated according to the present disclosure may have been subjected to standard tests or may have been identified, without examination, as one at risk due to the presence of one or more risk factors associated with the disease or condition.

[0348] In some embodiments, the subject has (e.g., confirmed by testing), or is suspected of having, an inflammatory condition. In some embodiments, the subject has an inflammatory condition. In some embodiments, the subject has been diagnosed with an inflammatory condition. In some embodiments, the subject is at risk of developing an inflammatory condition.

[0349] In some embodiments, the subject is a mammal. In some embodiments, the subject is a mammal selected from the group consisting of a dog, a cat, a mouse, a rat, a hamster, a guinea pig, a horse, a pig, a sheep, a cow, a chimpanzee, a macaque, a cynomolgus, and a human. In some embodiments, the subject is a primate. In some embodiments, the subject is a human.

[0350] In some embodiments, the subject has asthma, eczema (atopic dermatitis), food allergy, prurigo nodularis, chronic rhinosinusitis with nasal polyps, keloids, eosinophilic esophagitis, prostate cancer, chronic urticaria, bullous pemphigoid, localized scleroderma, alopecia areata, ulcerative colitis, aspirin-exacerbated respiratory disease, metastatic nonsmall cell lung cancer, Netherton syndrome, or a combination thereof.

[0351] In some embodiments, the subject is immune-compromised (e.g., has an underlying disorder or is on immunosuppressive therapy).

[0352] In some embodiments, the subject is 40 years or older, e.g., at least: 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 years old.

[0353] “A therapeutically effective amount,” “an effective amount” or “an effective dosage” is an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result (e.g., treatment, healing, inhibition or amelioration of physiological response or condition, etc.). The therapeutic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses.Thus, a therapeutically effective amount may be administered in one or more administrations. A therapeutically effective amount may vary according to factors such as disease state, age, sex, and weight of a mammal, mode of administration and the ability of a therapeutic, or combination of therapeutics, to elicit a desired response in an individual.

[0354] An effective amount of an agent to be administered can be determined by a clinician of ordinary skill using the guidance provided herein and other methods known in theart. Relevant factors include the given agent, the pharmaceutical formulation, the route of administration, the type of disease or disorder, the identity of the subject (e.g., age, sex, weight) or host being treated, and the like. For example, suitable dosages can be from about 0.001 mg / kg to about 100 mg / kg, from about 0.01 mg / kg to about 100 mg / kg, from about 0.01 mg / kg to about 10 mg / kg, from about 0.01 mg / kg to about 1 mg / kg body weight per treatment. Determining the dosage for a particular agent, subject and disease is well within the abilities of one of skill in the art. Preferably, the dosage does not cause or produces minimal adverse side effects.

[0355] Desired response or desired results include effects at the cellular level, tissue level, or clinical results. As such, “a therapeutically effective amount” or synonym thereto depends upon the context in which it is being applied. For example, in some embodiments it is an amount of the composition sufficient to achieve a treatment response as compared to the response obtained without administration of the composition. In some embodiments, it is an amount that results in a beneficial or desired result in a subject as compared to a control. As defined herein, a therapeutically effective amount of a composition disclosed herein may be readily determined by one of ordinary skill by routine methods known in the art. Dosage regimen and route of administration may be adjusted to provide the optimum therapeutic response.

[0356] In some embodiments, methods disclosed herein are used for prophylactic therapy. In some embodiments, the effective dosage is sufficient to prevent the subject from experiencing symptoms of an inflammatory condition.

[0357] In some embodiments, methods disclosed herein are used for treating an inflammatory condition e.g., atopic dermatitis).

[0358] The term “treating” or “treatment” refers to the medical management of a subject with the intent to improve, ameliorate, stabilize (i.e., not worsen), prevent or cure a disease, pathological condition, or disorder — such as the particular indications exemplified herein. This term includes active treatment (treatment directed to improve the disease, pathological condition, or disorder), causal treatment (treatment directed to the cause of the associated disease, pathological condition, or disorder), palliative treatment (treatment designed for the relief of symptoms), preventative treatment (treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder); and supportive treatment (treatment employed to supplement another therapy). Treatment also includes diminishment of the extent of the disease or condition; preventingspread of the disease or condition; delay or slowing the progress of the disease or condition; amelioration or palliation of the disease or condition; and remission (whether partial or total), whether detectable or undetectable. “Ameliorating” or “palliating” a disease or condition means that the extent and / or undesirable clinical manifestations of the disease, disorder, or condition are lessened and / or time course of the progression is slowed or lengthened, as compared to the extent or time course in the absence of treatment. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the condition or disorder, as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.

[0359] In some embodiments, the effective dosage is sufficient to reduce IL-4 / IL-13 signaling in the subject. In some embodiments, the reduction in IL-4 / IL-13 signaling is by at least about 10%, e.g., by at least about: 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. In some embodiments, the reduction in viral load is about 10-99%, e.g., about: 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0360] In some embodiments, the effective dosage is sufficient to inhibit binding of IL-4 or IL- 13 to IL-4Ra, target cells, or both. In some embodiments, the reduction in binding is by at least about 10%, e.g., by at least about: 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. In some embodiments, the reduction in binding is about 10-99%, e.g., about: 10-98%, 15-98%, 15-97%, 20-97%, 20-96%, 25-96%, 25-95%, 30-95%, 30-94%, 35-94%, 35-93%, 40-93%, 40-92%, 45-92%, 45-91%, 50-91%, 50-90%, 55-90%, 55-85%, 60-85%, 60-80%, 65-80%, 65-75%, or 70-75%.

[0361] A therapeutic agent described herein can be administered via a variety of routes of administration, including, for example, oral, dietary, topical, transdermal, rectal, parenteral (e.g., intra-arterial, intravenous, intramuscular, subcutaneous injection, intradermal injection), intravenous infusion and inhalation (e.g., intrabronchial, intranasal or oral inhalation, intranasal drops) routes of administration, depending on the compound and the particular disease to be treated. Administration can be local or systemic as indicated. The preferred mode of administration can vary depending on the particular compound chosen.

[0362] In some embodiments, a polypeptide, composition, or pharmaceutical composition disclosed herein is administered to a subject as a monotherapy.

[0363] In some embodiments, a polypeptide, composition, or pharmaceutical composition disclosed herein is administered to a subject in combination with one or more additional therapeutic agents (e.g., concurrently or sequentially with one or more additional therapeutic agents) or prophylactic agents (e.g., concurrently or sequentially with one or more prophylactic agents). In some embodiments, a subject has been previously treated with one or more therapeutic agents prior to being administered a polypeptide, composition, or pharmaceutical composition disclosed herein. In some embodiments, methods disclosed herein comprise administering a therapeutically effective amount of one or more additional therapeutic agents to the subject at the same time as, or following administration of a polypeptide, composition, or pharmaceutical composition disclosed herein. In some embodiments, methods disclosed herein comprise administering a therapeutically effective amount of one or more prophylactic agents to the subject before, at the same time as, or following administration of a polypeptide, composition, or pharmaceutical composition disclosed herein. In some embodiments, the subject previously received a therapeutic or prophylactic agent.

[0364] Non-limiting examples of additional therapeutic agents include an additional anti- IL-4Ra antibody or antigen-binding fragment thereof, an inhaled corticosteroid, a leukotriene modifier, a combination corticosteroid / long-acting beta agonist inhaler, a bronchodilator, a short-acting beta agonist, an anticholinergic agent, an oral or intravenous corticosteroid, a calcineurin inhibitor, an anti-inflammatory agent, an anti-rheumatic agent, an immunosuppressant, a biologic agent, an antihistamine, an epinephrine, an oral immunotherapy, a topical corticosteroid, a topical anesthetic, an immunomodulatory agent, a nasal corticosteroid, an antifungal agent, an anti-androgen agent, a luteinizing hormone- releasing hormone (LHRH) agonist, an gonadotropin-releasing hormone (GnRH) antagonist, a chemotherapy, a targeted drug therapy, a steroid-sparing agent, and combinations thereof.

[0365] Administration of the two or more therapeutic agents encompasses coadministration of the therapeutic agents in a substantially simultaneous manner, such as in a pharmaceutical combination. In some embodiments, such administration encompasses coadministration in multiple containers, or separate containers (e.g., capsules, powders, and liquids) for each therapeutic agent. Such administration also encompasses use of each type of therapeutic agent in a sequential manner, either at approximately the same time or at differenttimes. The composition described herein and the second therapeutic agent can be administered via the same administration route or via different administration routes.

[0366] In some embodiments, the disclosure provides methods of preventing one or more symptoms of an inflammatory condition in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable carrier and, wherein as an active ingredient, any one of polypeptides or fusion proteins described herein.

[0367] In some embodiments, the disclosure provides methods of treating an inflammatory condition in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable carrier and, wherein as an active ingredient, any polypeptide or fusion protein described herein.

[0368] In some embodiments, the disclosure provides methods of reducing one or more symptoms of an inflammatory condition in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable carrier and, wherein as an active ingredient, any polypeptide or fusion protein described herein.

[0369] In some embodiments, the disclosure provides methods of inhibiting binding of IL-4 or IL-13 to a target cell, comprising contacting the target cell an effective amount of any polypeptide or fusion protein described herein.

[0370] In some embodiments, the disclosure provides methods of inhibiting binding of IL-4 or IL- 13 to a target protein on a target cell, comprising contacting the target cell an effective amount of any polypeptide or fusion protein described herein.

[0371] Headings used in this application are for convenience only and do not affect the interpretation of this application.

[0372] Preferred features of each of the aspects or embodiments provided by the invention are applicable to all of the other aspects or embodiments of the invention mutatis mutandis and, without limitation, are exemplified by the dependent claims and also encompass combinations and permutations of individual features (e.g., elements, including numerical ranges and exemplary embodiments) of particular embodiments and aspects of the invention, including the working examples. For example, particular experimental parameters exemplified in the working examples can be adapted for use in the claimed invention piecemeal without departing from the invention. For example, for materials that are disclosed, while specific reference of each of the various individual and collectivecombinations and permutations of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein. Thus, if a class of elements A, B, and C are disclosed as well as a class of elements D, E, and F and an example of a combination of elements A-D is disclosed, then, even if each is not individually recited, each is individually and collectively contemplated. Thus, in this example, each of the combinations A-E, A-F, B- D, B-E, B-F, C-D, C-E, and C-F are specifically contemplated and should be considered disclosed from disclosure of A, B, and C; D, E, and F; and the example combination A-D. Likewise, any subset or combination of these is also specifically contemplated and disclosed. Thus, for example, the sub-groups of A-E, B-F, and C-E are specifically contemplated and should be considered disclosed from disclosure of A, B, and C; D, E, and F; and the example combination A-D. This concept applies to all aspects of this application, including elements of a composition of matter and steps of method of making or using the compositions.

[0373] The forgoing aspects of the invention, as recognized by the person having ordinary skill in the art following the teachings of the specification, can be claimed in any combination or permutation to the extent that they are novel and non-obvious over the prior art — thus, to the extent an element is described in one or more references known to the person having ordinary skill in the art, they may be excluded from the claimed invention by, inter alia, a negative proviso or disclaimer of the feature or combination of features.Table 1. Non-limiting examples of IL-4Rq sequencesTable 2, Variable Domain Amino Acid SequencesTable 3 CDR Amino Acid SequencesTable 4, Constant Domain Amino Acid SequencesTable 5, Heavy Chain Amino Acid SequencesTable 6, Light Chain Amino Acid SequencesComputational Definition and Verification of Sequences

[0374] Applicant, through computational design and experimental validation, has facilitated an understanding of structure and function interrelation of anti-IL-4Ra antibodies and herein provides the skilled artisan the means to use such understanding. For example, the Application provides Computer Program Listing Appendices (also referred to herein as Appendices A, B, and C, respectively) that can be used to both evaluate (score) a given sequence or generate a sequence having a score that, when evaluated, is above a given threshold. These Appendices include a Potts model of polypeptides provided by embodiments.

[0375] A statistical machine learning model, according to some embodiments, was developed to predict functional properties of polypeptides, e.g., antibody binding to an interleukin-4 receptor alpha (IL-4Ra) target. An example model was trained on experimental binding data and uses a Potts statistical model to make predictions about, e.g., whether novel antibody sequences will bind to IL-4Ra.

[0376] For model selection, multiple example candidate model architectures were evaluated as described below, and a Potts model was selected based on optimal performance across multiple metrics: a) Linear model (224 parameters) b) Uniform model (224 parameters) c) Positive Potts model (169,408 parameters) d) Mixture model (231,492 parameters) e) Joint model (246,717 parameters) f) Ratio model (383,128 parameters)

[0377] The selected Potts model, according to some embodiments, demonstrates strong predictive performance according to the following example model performance metrics: a) Area Under Curve (probability of distinguishing one randomly selected positive instance from one randomly selected negative instance): 0.831 b) Accuracy: 76.2% c) Precision (true positives / (true positives + false positives)): 60.5% d) Recall (true positives / (true positives + false negatives)): 84.3% e) Fl Score (harmonic mean of precision and recall): 0.704 [2 * ((0.605 * 0.843) / (0.605 + 0.843)) = -0.704]f) Binding Threshold: -27.87 (model score)

[0378] In some embodiments, the model may utilize an example dataset that is sourced from experimental binding affinity data from antibody variants for a target of interleukin-4 receptor alpha (IL-4Ra). The dataset may include 378 example antibody sequences with binding affinities measured to determine binding dissociation constants (KD values). An example binding threshold of log(Ko) = 0.375 may be established, z.e., sequences with log(Ko) < 0.375 may be classified as “binding.” Other binding threshold values are also suitable.

[0379] The model architecture may be a positive Potts model that employes a statistical physics-based approach for sequence modeling. 169,408 example model parameters may be used. The model may take as input variable regions from heavy chain (VH) and light chain (VL) antibody sequences. An example VH region input may include 25 amino acid positions, while an example VL region input may include 17 amino acid positions, resulting in a total of 42 example amino acid positions for analysis.

[0380] In some embodiments, for a statistical approach, the model may leverage a Potts statistical framework that analyzes single amino acid site effects. For example, the framework may capture a preference for specific amino acids at each amino acid position in a given sequence of a polypeptide. The framework may further analyze pairwise interactions between amino acids at different positions. For example, the framework may model epistatic effects between amino acid positions. The framework may further perform cross-entropy scoring. For example, the framework may use negative log-pseudolikelihood for sequence evaluation.

[0381] In certain embodiments, the model may utilize a scoring mechanism where higher scores indicate, e.g., a greater likelihood of binding. According to an embodiment, a threshold value of approximately -27.87 for negative log-pseudolikelihood may separate, e.g., binding from non-binding sequences. Other threshold values may also be used. As described herein, a scoring mechanism according to some embodiments may incorporate both individual amino acid preferences and inter-position correlations.

[0382] It should be noted that in certain embodiments, a Potts model may not directly predict KD or log(Ko) values, but rather may generate a score (e.g., a negative log- pseudolikelihood) that correlates with KD or log(Ko) values. According to some embodiments, a threshold for a Potts model score may be specified so as to maximally correctly classify a set of binders (e.g., log(Ko) < 0.375) from nonbinders (e.g., log(Ko) >=0.375) using training data. In certain embodiments, a threshold for a Potts model score may correspond either exactly or approximately to a log(Ko) value of 0.375.

[0383] The Computer Program Listing Appendices are referred to as Appendix A (score_concise.txt), Appendix B (score.txt), and Appendix C (fit_model.txt), which are herein incorporated by reference in their entireties. A person having ordinary skill in the art can recognize that Appendix A or Appendix B can respectively be renamed “score concise.py” or “score. py” to be executed using standard libraries (e.g., math, sys, etc.) and without external dependencies in a Python environment (e.g., Python 3.x), compiler, or other equivalent environment that can run Python scripts, and that Appendix C can be renamed “fit model.etab” and can be loaded by the script of Appendix A or Appendix B. A person having ordinary skill in the art can recognize that, in some embodiments, a Python environment may also run or compile the script of Appendix A or Appendix B without renaming. Other known programming languages and / or scripting environments are also suitable. When running the score_concise . py or score . py file, two sequences (or one combined sequence) can be inputted. In embodiments, the below description further describes the scripts of Appendices A and B and the model file of Appendix C.

[0384] The Potts model may be represented by a table of trained model parameters. The table may include single amino acid site parameters and pairwise amino acid parameters. For the single site parameters, the table may include a position of a residue (e.g., amino acid), the residue, and a value (e.g., a floating-point value) associated with the residue at that position. For the pairwise parameters, the table may include two positions, two residues, and a value (e.g., a floating-point value) associated with the two residues at the respective positions. The model parameters may be stored in the plaintext model file f it_model . etab.

[0385] In some embodiments, a script (e.g., Appendix A or Appendix B) may be employed to determine whether a sequence pair (e.g., VH and VL — although, for clarity, the sequence pair, in different embodiments, can be separate polypeptide chains or, in some embodiments, a single polypeptide chain, e.g., an scFV) is claimed under the model — z.e., whether the sequence pair, when scored by the model, is above a threshold. The script may be referred to as a computationally binding optimized (CBO) script. The script may take full- length sequences as inputs, e.g., VH and VL sequences provided as complete antibody sequences. In an embodiment, the sequences may be in standard single-letter amino acid code format; other known formats are also suitable. To continue, the script may then confirm thatthe input sequences match specific constant region templates. Shown below are example sequence templates according to an embodiment:Heavy Chain TemplateEVQLVESGGGLEQPGGSLRLSCAGSGXXXXXXAMTWVRQAPGKGLEWVSSIXGSGXXXYYADSVKGRFTISRDNS KNTLYLQMNSLRAEDTAVYYCXXXXXXXXXXPXYXGXXXWGQGTTVTVSSLight Chain TemplateDIVMTQSPLSLPVTPGEPASISCRSSXSXXYXXGXXYLDWYLQKSGQSPQLLIYXXXNRASGVPDRFSGSGSGTD FTLKI SRVEAEDVGFYYCXXXXXXPXTFGQGTKLE I K where 'X' denotes variable positions that are analyzed by the model.

[0386] In certain embodiments, a script, e.g., score_concise . py (Appendix A), may be used in either manner shown below:Command Line Usage python score_concise . py "VH_SEQUENCE" "VL_SEQUENCE"Python Application Programming Interface (API) Usage from score_concise import is_binding_antibody# Returns True if predicted to bind, False otherwise result = is_binding_antibody (vh_sequence , vl_sequence)

[0387] The script may construct two parameter hash tables for single site parameters and pairwise parameters, respectively, by loading an existing model file (e.g., Appendix C) and extracting single site amino acid positions / amino acid position pairs, amino acid identifiers, and corresponding values for each. For the single site parameters table, the script may extract one position, one residue identifier, and one value associated with the residue at the position. For the pairwise parameters table, the script may extract a first position, a second position, a first residue identifier, a second residue identifier, and a value associated with the first residue at the first position and the second residue at the second position.

[0388] The two sequences, the VH sequence and VL sequence, input to the script may then be aligned. The alignment may confirm that the VH sequence and VL sequence are the same length as template sequence(s) and that given sections of both input sequences are the same as the template sequence(s). Once confirmed, residues from both input sequences at specified positions may then be extracted to form a trimmed and concatenated sequence. Inturn, a sequence score may be determined for the given concatenated sequence using the two parameter hash tables.

[0389] The following steps describe some embodiments of determining the sequence score. The is_binding_antibody ( ) function of the script of Appendix A may call the calculate_binding_score ( ) function to determine and return a score for the sequence. The return value of the calculate_binding_score ( ) function may represent a sum of cross-entropy loss at each amino acid position in the concatenated sequence. If the sequence score is greater than the threshold, which in some embodiments is approximately -27.87, then the sequences meet the required parameters (e.g., are confirmed / verified by the Potts model) and the is_binding_antibody ( ) function may return true. Otherwise, the function may return false and the sequence pair is not confirmed by the model. When the is_binding_antibody ( ) function returns true, this may indicate that the sequence pair is suitable for the given purpose (e.g., a purpose related to binding to human IL-4Ra).

[0390] In some embodiments, the script of Appendix A may define a sequence scoring function (calculate_binding_score) that takes as inputs a sequence and a pair of parameter hash tables constructed from a model file (e.g., Appendix C). This function may determine a plurality of energy scores, including single amino acid energy scores and pairwise amino acid energy scores, for each amino acid position in the sequence. For a given amino acid position in the sequence, the calculate_binding_score function may use a Potts model to determine a single amino acid energy score and a sum of pairwise amino acid energy scores for each possible residue (e.g., amino acid) at the position. As described above, the function may check each possible residue at each position requested. A person of ordinary skill in the art can recognize that the script can replace residues at each given position (as in the script of Appendix A). In some embodiments, a similar script could add residues into the sequence. Continuing with the calculate_binding_score function, the single amino acid energy score and the sum of pairwise amino acid energy scores may be summed and added to an array of amino acid energies. A maximum energy value in the array of amino acid energies may then be determined and the array may be further used to generate a partition function. In some embodiments, a “softmax” function or “LogSumExp” (also called “RealSoftMax”) function may be applied to normalize the amino acid energies for numerical stability; other known normalization techniques are also suitable. A cross-entropymetric e.g., cross-entropy loss) may be determined for the given amino acid position based on an amino acid at the amino acid position, the maximum energy value, and the partition function. Other known metrics are also suitable. In turn, the calculate_binding_score function may sum the cross-entropy metrics at each amino acid position and use the resulting sum to generate a sequence score, which score may be returned by the function.

[0391] In some embodiments, the script of Appendix A may define a load_model_parameters ( ) function to load Potts model parameters from a model file (e.g., Appendix C). The file may be a f it_model . etab file in plaintext “.etab” format; other model file formats are also suitable, including text-based and binary formats, for nonlimiting examples. To continue, each line in the file having three items may be unpacked by the load_model_parameters ( ) function and converted into an entry in a first hash table for single amino acid site parameters. Each line in the file having five items may be unpacked and converted into an entry of a second hash table for pairwise amino acid parameters. The two hash tables may then be returned by the load_model_parameters ( ) function.

[0392] The above steps and functions relate to examples of a “computationally binding optimized” (CBO) sequence, amino acid sequence, or polypeptide sequence. Such a sequence is a sequence that, when inputted into the above CBO script of Appendix A, returns a value of true. A person having ordinary skill in the art can recognize that the script can be implemented in other ways and / or with other series of steps, e.g., in the form of the script of Appendix B. A person having ordinary skill in the art can also recognize that other functionally equivalent tables can be used in place of the table of Appendix C. However, the provided Python scripts and table, when executed by a processor, are configured to output whether a given polypeptide sequence is a CBO sequence. Therefore, any sequence that, when analyzed by the scripts, results in a Boolean “true” output, is considered a CBO sequence. Any sequence that, when run by the scripts, results in a Boolean “false” output, is considered excluded from the group of the CBO sequences. Therefore, in some embodiments, the scripts of Appendix A and Appendix B themselves do not determine CBO sequences, but instead are tools to confirm whether a given sequence is a CBO sequence or not. When a CBO sequence is an amino acid sequence, the corresponding molecule having the amino acid sequence is referred to as a CBO polypeptide.

[0393] In certain embodiments, a similar script to Appendix A or Appendix B can return calculated scores of a given or multiple sequences in addition to or separate from a Boolean value representing the scores exceeding a threshold.

[0394] In some embodiments, a similar script to Appendix A or Appendix B 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 sequence template. In some embodiments, a brute force technique can be applied using the calculate_binding_score function of Appendix A.

[0395] Shown below are example predictions by the script of Appendix A (score_concise . py) according to an embodiment:Binding Prediction (True) python score_concise . py " EVQLVESGGGLEQPGGSLRLSCAGSGQLDSSYAMTWVRQAPGKGLEWVSSINGSGLASYYA DSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCMQSLQTYFTFPRYDGYSSWGQGTTVTV SS" " DIVMTQSPLSLPVTPGEPAS I SCRSSNSTAYRDGRLYLDWYLJQKSGQSPQLLJIYTAANRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGFYYCVRYYLDPVTFGQGTKLEIK"Score = -19.85 (> threshold -27.87) Predicted to bindNon-binding Prediction (False) python score_concise . py " EVQLVESGGGLEQPGGSLRLSCAGSGQLDSYYAMTWVRQAPGKGLEWVSSINGSGLASYYA DSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCMQSLSTYFTFPSYDGYSGWGQGTTVTV SS" "DIVMTQSPLSLPVTPGEPASISCRSSNSTAYKDGALYLDWYLQKSGQSPQLLIYTIANRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGFYYCIRYYLDPVTFGQGTKLEIK"Score = -28.37 (< threshold -27.87) Predicted not to bind

[0396] The Potts model, according to some embodiments, was validated using multiple example techniques as described below:a) Threshold optimization'. Youden’s J statistic was used to find an optimal discrimination threshold. b) Cross-validation'. Model performance was assessed on held-out test sequences. c) Template matching'. Templates were utilized to ensure sequence compatibility before prediction.

[0397] Shown below are example validation results for the Potts model according to some embodiments: a) True Positive Rate. 84.3% (correctly identifies binding antibodies) b) False Positive Rate. 27.9% (incorrectly predicts binding for non-binding antibodies) c) True Negative Rate. 72.1% (correctly identifies non-binding antibodies) d) False Negative Rate. 15.7% (incorrectly predicts non-binding for binding antibodies)

[0398] In some embodiments, the scripts of Appendix A and Appendix B may depend on sequence templates by requiring input sequences to match specific constant region templates. The Potts model, according to some embodiments, may have a training data scope based on training the model on specific IL-4Ra binding data without validating generalization to other potential targets. Experimental validation may be performed on computational predictions generated by a model to confirm the predictions with experimental binding assays. The 27.9% false positive rate according to some embodiments discussed herein may indicate that approximately 1 in 4 predicted binders may not actually bind.

[0399] A Potts model according to an embodiment was applied to an example set of 17 test sequences, resulting in 12 of the 17 sequences (approx. 70.6%) being predicted as binding. Table 10 shows the 12 sequences predicted as binding and their corresponding log(Ko) scores.Table 10: Example Variable Sequences and log(Ko) ScoresExample Method Embodiment

[0400] FIG. 10 is a flowchart of a method 1000 for predicting a functional property of a polypeptide according to an embodiment. The method 1000 is computer-implemented and may be implemented using any computing device, e.g., a processor, or combination of computing devices known to those of skill in the art.

[0401] The method 1000 begins at step 1001 by, via a computationally binding optimized (CBO) model, for each amino acid position of an amino acid sequence of the polypeptide: (1) determining a plurality of energy scores based on the amino acid position in the amino acid sequence, (2) generating a partition function based on the plurality of energy scores determined, and (3) determining a cross-entropy metric (e.g., cross-entropy loss) based on (i) an amino acid at the amino acid position in the amino acid sequence, (ii) a maximum energy score of the plurality of energy scores determined, and (iii) the generated partition function. At step 1002, an analysis score of the polypeptide is generated based on each cross-entropy metric determined. The analysis score indicates a predicted functional property of thepolypeptide. The polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:37. The polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO: 5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.

[0402] In some example embodiments, the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) amino acid sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least about 70% amino acid sequence identity (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) amino acid sequence identity to SEQ ID NO:37. In some example embodiments, the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% amino acid sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% amino acid sequence identity to SEQ ID NO:37. In some example embodiments, the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:37.

[0403] As noted, the method 1000 is computer-implemented and, as such, the functionality and effective operations, e.g., the determining, generating, and determining (1001) and generating (1002), are automatically implemented by one or more digital processors. The method 1000 can also be implemented using any computer device or combination of computing devices known in the art. Among other examples, the method 1000 can be implemented using computer(s) / device(s) 50 and / or 60 described hereinbelow in relation to FIGs. 11 and 12.

[0404] In an example embodiment of the method 1000, a given plurality of energy scores determined at step 1001 may include at least one of: a single amino acid energy score and a pairwise amino acid energy score.

[0405] According to an example embodiment of the method 1000, for at least one amino acid position of the amino acid sequence of the polypeptide, determining the plurality of energy scores at step 1001 may be further based on having substituted the amino acid at the amino acid position in the amino acid sequence with each of a plurality of different amino acids.

[0406] In an example embodiment of the method 1000, for at least one amino acid position of the amino acid sequence of the polypeptide, generating the partition function at step 1001 may be further based on a softmax function.

[0407] According to an example embodiment of the method 1000, at least one of: (1) predicting the functional property of the polypeptide may be implementable by a script of Appendix A or Appendix B, and (2) the CBO model may be substantially similar to a table of Appendix C.

[0408] In an example embodiment of the method 1000, the generated analysis score at step 1002 may be above a threshold. The threshold may be a score from the CBO model of one or more of a reference polypeptide that includes a VH and VL pair selected from:SEQ ID NO:5 and SEQ ID NO:24 (Reference);SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6); SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7); SEQ ID NO: 13 and SEQ ID NO:32 (AB-8); SEQ ID NO: 14 and SEQ ID NO:33 (AB-9); SEQ ID NO: 15 and SEQ ID NO:34 (AB-10); SEQ ID NO: 16 and SEQ ID NO:35 (AB-11); SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12); SEQ ID NO: 18 and SEQ ID NO:37 (AB-13); SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c);SEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or a combination of any of the foregoing.

[0409] According to an example embodiment of the method 1000, the functional property predicted at step 1002 may be a binding affinity for interleukin-4 receptor alpha (IL-4Ra).

[0410] In an example embodiment of the method 1000, the functional property predicted at step 1002 may be at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 1 pM or less, a binding affinity for IL-4Ra characterized by a kaof about 100 x 105M_|s-1or less, a dissociation from IL-4Ra characterized by a kd of about 100 x 10’5s'1or less, a binding affinity for IL-4Ra characterized by an ECso of about 1 pM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 1 pM or less, a blocking activity against IL- 13 Type II signaling characterized by an ICso of about 1 pM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 1 pM or less.

[0411] According to an example embodiment of the method 1000, the functional property predicted at step 1002 may be at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.05 to 0.5 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7 x 105to 9 x 105T's-1or less, a dissociation from IL-4Ra characterized by a kd of about 2 x 10’5to 3 x 10’5s’ 1 or less, a binding affinity for IL-4Ra characterized by an EC50 of about 0.02 to 1.5 nM or less, a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 7.3 nM or less, a blocking activity against IL- 13 Type II signaling characterized by an IC50 of about 3 to 3.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by anIC50 of about 8 to 50 nM or less.

[0412] In an example embodiment of the method 1000, the functional property predicted at step 1002 may be at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.02 to 0.04 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7.7 x 105to 8.9 x 105M_|s_|or less, a dissociation from IL-4Ra characterized by a kd of about 2.1 x 10'5to 2.6 x 10'5s'1or less, a binding affinity for IL-4Ra characterized by an ECso of about 0.04 to 1.3 nM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 4.4 to 7.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 30 to 50 nM or less.

[0413] According to an example embodiment of the method 1000, the functional property predicted at step 1002 may be at least one of: a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 1.8 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 30 to 34 nM or less.

[0414] In an example embodiment of the method 1000, the functional property predicted at step 1002 may be an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 8 to 8.8 nM or less.

[0415] According to an example embodiment of the method 1000, the functional property predicted at step 1002 may relate to modulating activity of a target molecule. The target molecule may be interleukin-4 receptor alpha (IL-4Ra).

[0416] In an example embodiment of the method 1000, the analysis score generated at step 1002 may be a binding score.

[0417] According to an example embodiment of the method 1000, the functional property predicted at step 1002 may be a binding affinity.

[0418] In an example embodiment of the method 1000, the polypeptide may not comprise a heavy chain having an amino acid sequence that is identical to SEQ ID NO: 104 and a light chain having an amino acid sequence that is identical to SEQ ID NO: 161.

[0419] Embodiments, e.g., the method 1000, can be used as part of a design or development process. For instance, embodiments can be employed as part of an iterative design process where functional properties of candidate polypeptide sequences are evaluated. Based on results of the evaluation, e.g., a given candidate polypeptide sequence meeting a standard, a real-world polypeptide may be manufactured based on the polypeptide sequence that met the standard. In such an embodiment, a given polypeptide sequence may be automatically transmitted to a manufacturing device and the manufacturing device may responsively and automatically generate the real-world polypeptide.Computer Support

[0420] FIG. 11 is a schematic view of a computer network in which embodiments may be implemented. Client computer(s) / devices 50 and server computer(s) 60 provide processing, storage, and input / output (I / O) devices executing application programs and the like. Client computer(s) / device(s) 50 can also be linked through communications network 70 to other computing devices, including other client device(s) / processor(s) 50 and server computer(s) 60. The communications network 70 can be part of a remote access network, a global network (e.g., the Internet), cloud computing servers or service, a worldwide collection of computers, local area or wide area networks, and gateways that currently use respective protocols (e.g., TCP / IP, Bluetooth®, etc.) to communicate with one another. Other electronic device / computer network architectures are also suitable.

[0421] FIG. 12 is a block diagram illustrating an example embodiment of a computer node (e.g., client processor(s) / device(s) 50 or server computer(s) 60) in the computer network 70 of FIG. 11. Each computer node 50, 60 contains system bus 79, where a bus is a set of hardware lines used for data transfer among components of a computer or processing system. The system bus 79 is essentially a shared conduit that connects different elements of a computer system (e.g., processor, disk storage, memory, I / O ports, network ports, etc.) that enables transfer of information between the elements. Attached to the system bus 79 is an I / O devices interface 82 for connecting various input and output devices (e.g., keyboard, mouse, display(s), printer(s), speaker(s), etc.) to the computer node 50, 60. A network interface 86 allows the computer node to connect to various other devices attached to a network (e.g., the network 70 of FIG. 11). A memory 90 provides volatile storage for computer software instructions 92a and data 94a used to implement some embodiments of the present disclosure(e.g., the method 1000 of FIG. 10, etc.). A disk storage 95 provides non-volatile storage for the computer software instructions 92b and data 94b used to implement some embodiments of the present disclosure. A central processor unit 84 is also attached to the system bus 79 and provides for execution of computer instructions.

[0422] In an embodiment, the processor routines 92a-92b and data 94a-94b are a computer program product (generally referenced as 92), including a non-transitory, computer readable medium (e.g., a removable storage medium such as DVD-ROM(s), CD-ROM(s), diskette(s), tape(s), etc.) that provides at least a portion of the software instructions for the disclosure system. The computer program product 92 can be installed by any suitable software installation procedure, as is well known in the art. In another embodiment, at least a portion of the software instructions may also be downloaded over a cable, communication, and / or wireless connection. In other embodiments, the disclosure programs are a computer program propagated signal product embodied on a propagated signal on a propagation medium (e.g., a radio wave, an infrared wave, a laser wave, a sound wave, or an electrical wave propagated over a global network such as the Internet, or other network(s)). Such carrier medium or signals provide at least a portion of the software instructions for the present disclosure routines / program 92.

[0423] In alternative embodiments, the propagated signal is an analog carrier wave or digital signal carried on the propagated medium. For example, the propagated signal may be a digitized signal propagated over a global network (e.g., the Internet), a telecommunications network, or other networks (such as the network 70 of FIG. 11). In one embodiment, the propagated signal is a signal that is transmitted over the propagation medium over a period of time, such as the instructions for a software application sent in packets over a network over a period of milliseconds, seconds, minutes, or longer. In another embodiment, the computer readable medium of the computer program product 92 is a propagation medium that the computer system 50 may receive and read, such as by receiving the propagation medium and identifying a propagated signal embodied in the propagation medium, as described above for computer program propagated signal product.

[0424] Generally speaking, the term “carrier medium” or transient carrier encompasses the foregoing transient signals, propagated signals, propagated medium, storage medium, and the like.

[0425] In other embodiments, the program product 92 may be implemented as a so-called Software as a Service (SaaS), or other installation or communication supporting end-users.

[0426] Embodiments or aspects thereof may be implemented in the form of hardware including but not limited to hardware circuitry, firmware, or software. If implemented in software, the software may be stored on any non-transient computer readable medium that is configured to enable a processor to load the software or subsets of instructions thereof. The processor then executes the instructions and is configured to operate or cause an apparatus to operate in a manner as described herein.

[0427] Further, hardware, firmware, software, routines, or instructions may be described herein as performing certain actions and / or functions of the data processors. However, it should be appreciated that such descriptions contained herein are merely for convenience and that such actions in fact result from computing devices, processors, controllers, or other devices executing the firmware, software, routines, instructions, etc.

[0428] It should be understood that the flow diagrams, block diagrams, and network diagrams may include more or fewer elements, be arranged differently, or be represented differently. But it further should be understood that certain implementations may dictate the block and network diagrams and the number of block and network diagrams illustrating the execution of the embodiments be implemented in a particular way.

[0429] Accordingly, further embodiments may also be implemented in a variety of computer architectures, physical, virtual, cloud computers, and / or some combination thereof, and, thus, the data processors described herein are intended for purposes of illustration only and not as a limitation of the embodiments.EXEMPLIFICATIONExample 1. Generation and Characterization of IL-4Ra-Binding Polypeptides (Overview)Diversity Generation Campaign

[0430] The diversity generation campaign chose a human anti-IL-4Ra antibody as a Reference Antibody (“Reference”). A variant set of 384 antibodies was generated, and data on target binding (surface plasmon resonance or “SPR”) was acquired. A subset of 205 target-binding antibodies was selected and tested for developability and function. Two of the 205 target-binding antibodies bound IL-4Ra with high affinity and were selected as seeds for the co-optimization campaign.Co-Optimization Campaign

[0431] Using seeds from the diversity generation campaign, a second variant set of 384 antibodies was generated in the project learning and co-optimization campaign, and data on target binding was acquired. A subset of 163 target-binding antibodies was selected and tested for developability and function. Developability and function data were combined to identify 20 screening hits with acceptable developability parameters (Affinity-Capture SelfInteraction Nanoparticle Spectroscopy (AC-SINS) < 10 shift, size-exclusion chromatography (SEC or aSEC) > 85% monomer, a poly-specificity reagent binding via dissociation-enhanced lanthanide fluorescence immunoassay (PSR-DELFIA or PSR) < 10 fold-change, hydrophobic interaction chromatography (HIC or aHIC) < 10 minutes retention time) and similar binding kinetics relative to the Reference Antibody.Affinity Maturation Campaign

[0432] A set of 164 variants was screened of which 16 were further selected for functional screening. No variants exhibited improved functionality.Example 2. Binding AssaysSurface Plasmon Resonance (SPR)

[0433] Surface plasmon resonance (SPR) was employed to determine the kinetics and affinities of binding of a panel of interleukin 4 receptor subunit alpha (IL-4Ra) antibodies to human IL-4Ra. The Carterra LSA and Cytiva Biacore 8K+ instruments are label-free platforms that use SPR to measure binding interactions in real-time. Kinetic and affinity parameters were extracted from experimental data by an iterative process that found the best fit for a set of equations describing the interaction. The association rate constant ka (M^s'1) describes the rate at which a complex was formed. The dissociation rate constant kd (s'1) describes the rate at which a complex dissociated. The equilibrium dissociation constant KD (M) describes the strength of the interaction.Carterra LSA Non-Regenerative Capture Kinetics

[0434] To evaluate the affinities of binding of a panel of anti-IL-4Ra antibodies to human IL-4Ra at 25 °C, a non-regenerative capture kinetics approach was performed using a Carterra LSA instrument (Carterra, Salt Lake City, UT). A goat anti-human polyclonal antibody lawnwas prepared via amine-coupling onto a poly carboxylate hydrogel (HC30M) sensor chip using a single flow cell (SFC) to attain approximately 5,000 response units (RU). Each antibody was prepared at 8 pg / mL and captured onto regions of interest (ROI) for 15 min using four serial dockings of a 96-channel printhead (96PH) to create a 384-antibody array. Human IL-4Ra was prepared in HBSTE + BSA (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.01% Tween 20, 0.5 mg / mL BSA) running buffer and injected over the captured antibody array surface at nine concentrations in a 3 -fold serial dilution series from 1000 nM to 0.2 nM. Each analyte concentration was injected sequentially from low to high concentration in a single cycle using the SFC with a 5-min association phase and a 10-min dissociation phase. The surface was regenerated by injecting two pulses of 0.85% phosphoric acid for 15 seconds between each cycle. The assay was performed at 25°C with technical replicates. Kinetic parameters for the concentration series were obtained by double referencing and globally fitting the data to a 1 : 1 binding model with the 5% kd option selected using the Kinetics analysis software (Carterra). The affinities of human IL-4Ra binding to a panel of IL-4Ra antibodies were reported, and the results were expressed as the average ± standard deviation of one independent experiment with technical replicates (Table 7).Table 7, Carterra LSA Non-regenerative Capture Kinetics.Biacore 8K+ Multi-Cycle Capture Kinetics

[0435] To evaluate the kinetics of bindings of select anti-IL-4Ra antibodies to human IL- 4Ra at 25 °C, a multi-cycle capture kinetics approach was implemented using a Biacore 8K± instrument (Cytiva, Marlborough, MA). A goat anti-human polyclonal antibody surface was prepared via amine-coupling onto a carboxymethylated dextran (CM4) sensor chip to attainapproximately 5,000 response units (RU). Each antibody was prepared at 2-9 pg / mL and captured for 30 s at a flow rate of 10 pL min1to achieve a capture level of approximately 150 RU. Human IL-4Ra was prepared in HBS-EP+ (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, and 0.05% v / v surfactant P20) running buffer and injected over the captured antibody surface at six concentrations in a 3-fold serial dilution series from 30 nM to 0.1 nM. Each analyte concentration was injected in a separate cycle for 180 seconds at a flow rate of 30 pL min and the complex was allowed to dissociate for 3,600 seconds. The surface was regenerated by injecting two pulses of 0.85% phosphoric acid for 20 seconds at a flow rate of 30 pL min1in between each cycle. The assay was performed at 25 °C with technical replicates. Kinetic parameters for the concentration series were obtained by double referencing and globally fitting the data to a 1 : 1 binding model using the Biacore Insight Evaluation software (Cytiva). The kinetics and affinities of human IL-4Ra binding to select IL-4Ra antibodies were reported, and results were expressed from a global fit of replicate data of one independent experiment (Table 8).Table 8, Multi-Cycle Kinetics with Sample as Ligand (Biacore 8K+),LS = IgG4 LS variantYTE = IgG4 YTE variantExample 3. Functional AssaysHuman IL-4Ra Binding Assay (Indirect ELISA)

[0436] Binding affinity of antibodies to human IL-4Ra was assessed using an indirect enzyme-linked immunosorbent assay (ELISA). A 96-well half-area plate was coated with 2 pg / mL of recombinant human IL-4Ra (BioLegend, San Diego, CA) overnight. The next day, the plate was washed three times with ELISA Wash Buffer (phosphate-buffered saline (PBS)- Tween 0.5%), blocked with lx ELISA diluent (BioLegend) for one hour. The plates were then washed three times with ELISA Wash Buffer, followed by addition of the anti-IL-4Ra antibodies (1 :5 serial dilution 8-point curve starting at 5pg / mL antibody) and incubated at 37°C for one hour. The plate was washed three times with ELISA Wash Buffer. Horseradishperoxidase (HRP)-conjugated anti-human immunoglobulin G fragment crystallizable region (IgG Fc) secondary antibody (Promega Corporation, Madison, WI) at 1 :2000 dilution in lx ELISA diluent was added to detect binding of the primary antibody to the antigen. The plate was incubated for 30 minutes at 37°C with the secondary detection antibody, then the plate was washed and developed with 3,3',5,5'-tetramethylbenzidine (TMB) substrate solution (Thermo Fisher Scientific, Waltham, MA). The HRP enzymatic reaction was stopped after about 1 minute with acidic stop solution (Thermo Fisher Scientific), and the absorbance at 450 nm (A450) was determined using a standard plate reader (FIGs. 4A-4C).HEK-Blue Blocking Assay

[0437] The ability of an antibody to block binding of IL-4 / 13 to IL-4Ra was assessed using the HEK (human embryonic kidney)-Blue colorimetric assay. HEK-Blue cells are an engineered cell line developed by InvivoGen (San Diego, CA); these modified HEK cells have the gene for secreted alkaline phosphatase (SEAP) inserted within the locus activated by Signal Transducer and Activator of Transcription 6 (STAT6) downstream of IL-4 / IL-4Ra binding. When SEAP interacts with InvivoGen’ s QUANTLBlue solution, a reaction turns the liquid from pink to blue.

[0438] For this assay, HEK-Blue cells were grown to confluency in a media comprising DMEM + GLUTAMAX® (Gibco, Waltham, MA), 10% fetal bovine serum (FBS), 1% Penicillin-Streptomycin (Gibco), and 100 pg / mL Normocin (InvivoGen). Cells were removed from their flasks using 0.25% trypsin-EDTA (Gibco), washed in media, and replated in a 96- well flat-bottom plate at 1 million cells per milliliter, and 100 pl per well. The cells were then blocked with the anti-IL-4Ra antibody at a concentration curve for thirty minutes at 37°C. Afterwards, they were incubated overnight with lOOpl of 5 ng / mL human IL-4 / 13 cytokine (ACROBiosystems, Newark, DE).

[0439] The next day, 20 pl of cell supernatant was extracted from each well and mixed with 180 pl of QUANTLBlue solution (InvivoGen). This mixture was then allowed to develop in the dark for thirty minutes at room temperature. After developing, the absorbance was measured on a plate reader at 635 nm, indicating the amount of SEAP produced and thus the level of IL-4 / IL-4Ra binding in the cells despite antibody blocking (FIGs. 5A-5C).IL-4Ra Human Primary B Cell-based Assay (CD23 Inhibition)

[0440] Frozen peripheral blood mononuclear cells (PBMCs) from Donors were thawed, and B cells were isolated from frozen cell samples using the B cell isolation kit from STEMCELL Technologies (Vancouver, Canada; Cat. No. 17954) following the manufacturer’s instructions. A sample from each donor was taken and stained using the B cell flow panel listed in the table below to check purity of the B cell population.* Becton, Dickinson and Company (BD), Franklin Lakes, NJ

[0441] After isolation, B cells were plated at 3 x io5cells in 100 pL CIO media in a 96 well U-bottom plate and rested overnight at 37°C. The following day, anti-IL-4Ra antibody was added in 50 pL (3-fold 10-point curve starting at 150 pg / mL antibody) and incubated for 30 minutes at 37°C. IL-4 cytokine (BioLegend Cat. No. 574006) was added in 50 pL of media for controls to a final concentration of cytokine of 10 ng / mL in the assay. Total assay volume was 200 pL and was incubated for 48 hours. After 48 hours, the cells were stained according to the staining protocol in the table below to identify CD23 inhibition by anti-IL- 4Ra antibodies.- I l l -

[0442] Inhibition of CD23 expression was reported from 4 donors as IC50 (Table 9) and percent inhibition (FIGs. 6A-6D).Table 9. Inhibition of CD23 Expression by Primary B cells (IC50)LS = IgG4 LS variantYTE = IgG4 YTE variantRamos CD23 Inhibition Assay

[0443] Ramos (RAI) cells (ATCC, Manassas, VA; Cat. No. CRL-1596) were harvested when the cell density was greater than 3 * 106cells / mL. Ramos cells were plated at 3 * 105cells in 100 pL RPMI 1640 with 10% FBS and 1% Pen Strep media in a 96 well U-bottom plate. Anti-IL-4Ra antibody was added in 50 pL (3-fold 8-point curve starting at 30 pg / mL antibody) and was incubated for 30 minutes at 37°C. 50 pL IL-4 cytokine (BioLegend Cat. No. 574006) was added for a final concentration of 10 ng / mL of cytokine in a total assay volume of 200 pL. In control experiments, 50 pL media was added. The assay was incubated for 24 hours. After 24 hours, the cells were stained according to the protocol set forth in the tables below to identify CD23 inhibition by anti-IL-4Ra antibodies.

[0444] Results were reported as percent inhibition (FIG. 7).Example 4. Translational Assays pSTAT6 Whole Blood Assay

[0445] The objective was to evaluate IL-4Ra target engagement in primary cells via functional inhibition of relevant atopic dermatitis (AD) biomarkers, such as phosphorylated Signal Transducer and Activator of Transcription 6 (pSTAT6), and Thymus- and Activation- Regulated Chemokine (TARC or CCL17). Assay development, optimization, and proof-of- concept were done using the Reference (anti-IL-4Ra mAb) in peripheral blood mononuclear cells (PBMCs) and ultimately transferred into whole blood across three distinct donors.

[0446] See tables below for a list of materials used in this assay.*R&D Systems, Minneapolis, MN**eBioscience, Waltham, MA***BD Biosciences, Franklin Lakes, NJ

[0447] A volume of 180 pL / well of whole blood was plated in RPMI-1% FBS, in a 96 well flat bottom plate. Appropriate concentration of mAb was added at 20x (in a volume of 10 pL). The plate was incubated at 37°C for 20 minutes. Either 10 ng / mL or 1 ng / mL of rh- IL-4 was then added in 20x (volume of lOpL). The plate was incubated at 10 minutes at 37°C. The plate was centrifuged, and the supernatant was aspirated to leave a volume of about 100 pL.

[0448] Stimulation was then stopped by addition of Lyse / Fix buffer (BD) at a volume of 20 times the remaining volume of the blood. The samples were vigorously mixed via pipetting and then incubated at 37°C for 10-15 minutes during lysing of red blood cells (RBCs) and fixation. The cells were then washed twice in flow cytometry staining buffer and stained with fixable viability dye and FC block for 15 minutes at 4°C. Extracellular markers were added according to the concentrations suggested by manufacturer’s instructions. The cells were then washed 2x with flow cytometry staining buffer before being resuspended in BD Phosflow Perm II buffer for 30 minutes. The cells were once again washed as describedpreviously and resuspended in intracellular staining markers (pSTAT6) according to manufacturer’s instructions for 30 minutes at room temperature. After final 2 washes, the cells were resuspended in flow cytometry staining buffer, run on Attune CytPix flow cytometer (Invitrogen), and analyzed using FlowJo software (FlowJo, Ashland, OR). The graphs of data exported from FlowJo were visualized and statistically analyzed using GraphPad Prism (GraphPad Software, Boston, MA).

[0449] In the presence of IL-4, there was a dose-dependent decrease in pSTAT6 expression with both the Reference (hIgG4) and AB-13c (hIgG4) in both B cells (CD19+) and T cells (CD4+) (FIGs. 8 A, 8B). These results were consistent with the data obtained from two additional whole blood donors (FIGs. 8C, 8D). The dose-dependent decreases in pSTAT6 were comparable between the Reference and AB-13c in whole blood. Minimal impact from the biological matrix of whole blood (matrix effect) was observed as evidenced by the ability of the respective antibodies to retain pSTAT6 modulation, indicative of functional inhibition of IL-4Ra.TARC release (from PBMCs) assay

[0450] The objective was to evaluate IL-4Ra target engagement in primary cells via functional inhibition of relevant atopic dermatitis (AD) biomarkers, such as phosphorylated Signal Transducer and Activator of Transcription 6 (pSTAT6) and Thymus- and Activation- Regulated Chemokine (TARC or CCL17). TARC release can be used as a functional readout of anti-IL-4Ra activity via ELISA readout. Assay development, optimization and proof-of- concept experiments were done using the Reference (hIgG4 IL-4Ra mAb) in PBMCs. AB- 13c (hIgG4) was tested in comparison to the Reference in one donor. The assay was optimized to be used in further potential uses of AB- 13 c.

[0451] See table below for a list of materials used in this assay.

[0452] PBMCs were plated in a 96 well flat bottom plate at about 5* 104to about 1 * 105cells per well in a volume of 50 pL of RPMI-10% FBS. An appropriate concentration of mAb was added at 2x (in a volume of 50pL). The plate was incubated at 37°C for 20 minutes.Zero, 1, or 10 ng / mL of rh-IL-4 was then added in 3x (volume of 50 pL). The plate was then incubated at 37°C for 48-72 hours and then centrifuged. The supernatants were collected and frozen at -80°C until use. Undiluted samples were run in accordance with manufacturer’s instructions for the Human CCL17 / TARC QUANTIKINE® ELISA kit (R&D Systems). Optical density (OD) of the samples were read at 450 nM on a spectrophotometer. The data was analyzed using a nonlinear regression curve and graphed using GraphPad Prism software.

[0453] A dose-dependent induction of TARC release in response to rh-IL-4 stimulation was observed (FIG. 9A). An ideal range for IL-4 stimulation was from 0.1 to 100 ng / mL. At different levels of IL-4 stimulation (1, 10, and 100 ng / mL IL-4), AB-13c showed a degree of TARC suppression comparable to the Reference (FIGs. 9B-9D).

[0454] The teachings of all patents, published applications, and references cited herein are incorporated by reference in their entirety.

[0455] While example embodiments have been particularly shown and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the embodiments encompassed by the appended claims.APPENDIX A - “SCORE CONCISE.PY” import math# Model parameters and amino acid constants BINDING_THRESHOLD = -27.869728088378906 AMINO_ACIDS = ' ACDEFGHIKLMNPQRSTVWY- '# Antibody templates with variable regions marked as 'X' VH_TEM PLATE =' EVQLVESGGGLEQPGGSLRLSCAGSGXXXXXXAMTWVRQAPGKGLEWVSSIXGSGXXXYYADSVKGRFTI SRDNSKNTLYLQMNSLRAEDTAVYYCXXXXXXXXXXPXYXGXXXWGQGTTVTVSS 'VL_TEM PLATE =' DIVMTQSPLSLPVTPGEPASISCRSSXSXXYXXGXXYLDWYLQKSGQSPQLLIYXXXNRASGVPDRFSGS GSGTDFTLKISRVEAEDVGFYYCXXXXXXPXTFGQGTKLEIK '# Positions of variable regions in the templates VH_VARIABLE_POSITIONS = [26,27,28,29,30,31,51,55,56,57,96,97,98,99,100,101,102,103,104,105,107, 109, 111, 112, 113]VL_VARIABLE_POSITIONS =[26,28,29,31,32,34,35,54,55,56,93,94,95,96,97,98,100]# Amino acid code mappingdef load_model_parameters ( ) :"""Load single-site and pairwise interaction parameters from model file" " " single_site_params = { } pairwise_params = {} with open ( ' fit_model . etab ' ) as model_file: for line in model_file: parts = line. split () if len (parts) == 3: position, amino_acid, value = int (parts [0] [1:] ) , AA_CODE_MAP [parts [1] ] , -float (parts [2] ) single_site_params [ (position, amino_acid) ] = value elif len(parts) == 5: posl, pos2 = int (parts [0] [1 : ] ) , int (parts [1] [1 : ] ) aal, aa2 = AA_CODE_MAP [parts

[0002] ] , AA_CODE_MAP [parts [3] ] value = -float (parts [4] )# Store both orientations for symmetric interactions pairwise_params [ (posl , pos2, aal, aa2)] = value pairwise_params [ (pos2 , posl, aa2, aal) ] = value return single_site_params , pairwise_params def extract_variable_regions (vh_sequence , vl_sequence) :"""Extract variable region sequences if templates match""" vh_matches_template = (len (vh_sequence) == len (VH_TEMPLATE) and all(seq_aa == template_aa for seq_aa, template_aa in zip (vh_sequence , VH_TEMPLATE) if template_aa != 'X' ) )vl_matches_template = (len (vl_sequence ) == len (VL_TEMPLATE) and all(seq_aa == template_aa for seq_aa, template_aa in zip (vl_sequence , VL_TEMPLATE) if template_aa != 'X' ) ) if not (vh_matches_template and vl_matches_template) : return None, None vh_variable = ' ' . j oin ( vh_sequence [pos] for pos in VH_VARIABLE_POSITIONS ) vl_variable = ' ' . j oin ( vl_sequence [pos] for pos in VL_VARIABLE_POSITIONS ) return vh_variable, vl_variable def calculate_binding_score (combined_sequence , single_site_params , pairwise_params) :"""Calculate binding score using statistical model""" total_loss = 0 for position in range (len (combined_sequence) ) : current_amino_acid = combined_sequence [position]# Calculate energy for each possible amino acid at this position amino_acid_energies = [] for amino_acid in AMINO_ACIDS: single_energy = single_site_params . get ( (position, amino_acid) , 0) pairwise_energy = sum (pairwise_params . get ( (position, other_pos, amino_acid, combined_sequence [other_pos] ) , 0) for other_pos in range (len (combined_sequence) ) if other_pos != position) amino_acid_energies . append (single_energy + pairwise_energy )# Apply softmax normalization for numerical stability max_energy = max (amino_acid_energies) normalized_energies = [energy - max_energy for energy in amino_acid_energies] partition_function = sum (math. exp (energy) for energy in normal ized_energies)# Calculate cross-entropy loss for current position current_aa_index = AMINO_ACIDS . index (current_amino_acid) position_loss = ( -amino_acid_energies [current_aa_index] + max_energy + math . log (partition_function) ) total_loss += position_loss return -total_loss def is_binding_antibody (vh_sequence , vl_sequence) :"""Predict if antibody sequences bind to IL4Ra target""" vh_variable, vl_variable = extract_variable_regions (vh_sequence , vl_sequence)if vh_variable is None or vl_variable is None: return False single_site_params , pairwise_params = load_model_parameters ( ) combined_variable_sequence = vh_variable + vl_variable binding_score = calculate_binding_score (combined_variable_sequence , single_site_params , pairwise_params) return binding_score > BINDING_THRESHOLD if > name == " main " : import sys if len (sys . argv) == 3: print (is_binding_antibody (sys . argv [1] , sys. argv [2] ) )5708.1079002 APPENDIX B – “SCORE.PY” import itertools import argparse import numpy as np #MAGIC NUMBERS THRESHOLD = -27.869728088378906 VH_CONST = 'EVQLVESGGGLEQPGGSLRLSCAGSGXXXXXXAMTWVRQAPGKGLEWVSSIXGSGXXXYYADSVKGRFTI SRDNSKNTLYLQMNSLRAEDTAVYYCXXXXXXXXXXPXYXGXXXWGQGTTVTVSS' VL_CONST = 'DIVMTQSPLSLPVTPGEPASISCRSSXSXXYXXGXXYLDWYLQKSGQSPQLLIYXXXNRASGVPDRFSGS GSGTDFTLKISRVEAEDVGFYYCXXXXXXPXTFGQGTKLEIK' VH_POSITIONS = [26, 27, 28, 29, 30, 31, 51, 55, 56, 57, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 107, 109, 111, 112, 113] VL_POSITION 26, 28, 29, 31, 32, 34, 35, 54, 55, 56, 93, 94, 95, 96, 97, 98, 100 ETAB_FILE =_model.etab' tripleToSingle = {def read_etab(etab_file:str) -> (dict,dict): '''load an etab file and produce two hash tables''' h = dict() j = dict() with open(etab_file,'r') as file: for line in file: items = line.replace('\n','').split(' ') if len(items) == 3: #Unpack i1, r1, v = items #Transform i1 = int(i1[1:]) r1 = tripleToSingle[r1] v = -float(v) # Negate like PottsModel does - 120 - 4178026.v1#Create New Entry in the hash table h [ (il, rl) ] = v elif len(items) == 5:#Unpack il, i2 , rl , r2 , v = items#Transform11 = int (il [1 : ] )12 = int (i2 [1 : ] ) rl = tripleToSingle [rl] r2 = tripleToSingle [r2] v = -float (v) # Negate like PottsModel does#Create New Entry in the hash table (symmetric) j [ (il , i2 , rl , r2 ) ] = v j [ (i2 , il , r2 , rl ) ] = v return h,j def trim_sequence ( sequence , positions, constant) : if len (sequence) != len (constant ) : return None elif not all ( [r == n for r, n in zip (list (constant ) , list (sequence) ) if r ! = 'X' ] ) : return None else : return j oin ( [sequence [p] for p in positions] ) def sequence_energy (h : diet , j zdict, sequence : str , tokens= ' ACDEFGHIKLMNPQRSTVWY- ' ) -> float: score = 0 #Score First Order Contributions for i,s in enumerate (sequence) : try: score += h [ (i , s) ] except KeyError: pass#Score Second Order Contributions for cl, c2 in itertools . combinations (enumerate (sequence) , 2 ) : assert cl [0] <= c2 [0] , 'first element of sequence should come before the second! ' try: score += j [ (cl [0] , c2 [0] , cl [1] , c2 [1] ) ] except KeyError: pass return score def score_sequence (h : diet , j :dict, sequence : str , tokens= ' ACDEFGHIKLMNPQRSTVWY- ' ) -> float:"""Compute negative log pseudolikelihood of sequence given Potts model parameters.""" total loss = 0.0 for position in range ( len ( sequence ) ) :# Compute logits for all possible tokens at this position logits = [] for token in tokens: logit = h . get ( (position, token) , 0.0)# Add coupling contributions from all other positions for other_pos in range (len (sequence) ) : if other_pos != position: logit += j . get ( (position, other_pos, token, sequence [other_pos] ) , 0.0) logits . append ( logit )# Cross-entropy loss: -log (softmax (logits) [native_token] ) native_idx = tokens . index ( sequence [position] ) max_logit = max (logits) log_sum_exp = max_logit + np . log (sum (np . exp (logit - max_logit) for logit in logits) ) total_loss += -logits [native_idx] + log_sum_exp return total_loss def check_sequence (vh : str , vl:str) -> bool:' ' ' take in a vh and vl sequence and return a boolean value either claiming or not claiming a sequence' ' ' h,j = read_etab (ETAB_FILE)# Module for Sequence Alignment: tvh = trim_sequence (vh, VH_P0SITI0NS, VH_C0NST) if tvh is None: return False tvl = trim_sequence (vl , VL_P0SITI0NS, VL_C0NST) if tvl is None: return False sequence = tvh+tvl#Score Aligned Sequence & Claim score = -score_sequence (h, j , sequence) if score > THRESHOLD: return True else : return False if > name == " main " : parser = argparse . Argument Parser ( ) parser . add_argument ( "vh" , type=str, help="a vh sequence to check if it is claimed") parser . add_argument ( "vl " , type=str, help="a vl sequence to check if it is claimed") args = parser . parse_args ( ) print (check_sequence (args . vh, args . vl ) )APPENDIX C - “FIT MODEL.ETAB” ALA 0.895677 CYS 0.895677 ASP 0.895677 GLU -4. 3349 PHE 0.895677 GLY 0.895677 HIS 0.895677 ILE 0.895677 LYS -3.00118 LEU 0.895677 MET 0.895677 ASN 0.895677 PRO 0.895677 GLN -5.54801 ARG 0.895677 SER -2.24382 THR 0.895677 VAL 0.895677 TRP 0.895677 TYR 0.895677 - 0.895677 ALA 0.922948 CYS 0.922948 ASP 0.922948 GLU -2.92475 PHE 0.922948 GLY 0.922948 HIS 0.922948 ILE 0.922948 LYS 0.922948 LEU -5.11871 MET 0.922948 ASN 0.922948 PRO 0.922948 GLN 0.922948 ARG 0.922948 SER -2.91924 THR 0.922948 VAL -4.72743 TRP 0.922948 TYR 0.922948 - 0.922948 ALA -1.26169 CYS 1.46015 ASP -3.04872 GLU 1.46015 PHE -1.9902 GLY 1.46015 HIS 1.46015 ILE 1.46015 LYS 1.46015 LEU -4.40878 MET 1.46015 ASN 1.46015 PRO 1.46015GLN -1.92623 ARG -3.05488 SER -1. 2104 THR 1.46015 VAL -1.87036 TRP 1.46015 TYR 1.46015 - 1.46015 ALA 0.538396 CYS 0.538396 ASP 0.538396 GLU 0.538396 PHE 0.538396 GLY 0.538396 HIS 0.538396 ILE 0.538396 LYS 0.538396 LEU 0.538396 MET 0.538396 ASN 0.538396 PRO 0.538396 GLN 0.538396 ARG 0.538396 SER -7.00321 THR -3.22632 VAL 0.538396 TRP 0.538396 TYR 0.538396 - 0.538396 ALA 1.754 CYS 1.754 ASP 1.754 GLU -2.82358 PHE 1.754 GLY 1.754 HIS -0.989874 ILE -2.6613 LYS 1.13113 LEU -1.4663 MET 1.754 ASN 1.754 PRO 1.754 GLN -0.324618 ARG -1.45489 SER -2.42668 THR -2.36249 VAL -3.39792 TRP 1.754 TYR -0.763516 - 1.754 ALA 1.09938 CYS 1.09938 ASP 1.09938 GLU 1.09938 PHE 1.09938 GLY 1.09938 HIS -2.92874ILE 1.09938 LYS -4.39023 LEU 1.09938 MET 1.09938 ASN -2.61388 PRO 1.09938 GLN 1.09938 ARG 1.09938 SER 1.09938 THR 1.09938 VAL -2.55157 TRP 1.09938 TYR -5.10569 - 1.09938 ALA 0.928653 CYS -2.92237 ASP -3.86799 GLU 0.928653 PHE 0.928653 GLY 0.928653 HIS 0.928653 ILE 0.928653 LYS 0.928653 LEU 0.928653 MET 0.928653 ASN -5. 2817 PRO 0.928653 GLN 0.928653 ARG 0.928653 SER -3.56856 THR 0.928653 VAL 0.928653 TRP 0.928653 TYR 0.928653 --- 0.928653 ALA -2.92528 CYS -2.12846 ASP 1.38191 GLU -2.07173 PHE 1.38191 GLY 1.38191 HIS 1.38191 ILE 1.38191 LYS -2.82335 LEU -4.16216 MET 1.38191 ASN 1.38191 PRO 1.38191 GLN 1.38191 ARG -3.22053 SER 1.38191 THR -2.01523 VAL 1.38191 TRP 1.38191 TYR 1.38191 - 1.38191 ALA -5.20899S8 CYS 0.998976S8 ASP 0.998976S8 GLU 0.998976S8 PHE 0.998976S8 GLY -3.95852S8 HIS 0.998976S8 ILE 0.998976S8 LYS 0.998976S8 LEU 0.998976S8 MET 0.998976S8 ASN 0.998976S8 PRO 0.998976S8 GLN 0.998976S8 ARG 0.998976S8 SER 0.998976S8 THR -4.19177S8 VAL -3.62332S8 TRP 0.998976S8 TYR 0.99897658 --- 0.99897659 ALA 0.921314S9 CYS 0.921314S9 ASP -4.58953S9 GLU 0.921314S9 PHE 0.921314S9 GLY 0.921314S9 HIS -2.9006S9 ILE 0.921314S9 LYS 0.921314S9 LEU 0.921314S9 MET 0.921314S9 ASN 0.921314S9 PRO 0.921314S9 GLN -2.84776S9 ARG 0.921314S9 SER -5.32445S9 THR 0.921314S9 VAL 0.921314S9 TRP 0.921314S9 TYR 0.921314S9 - 0.921314S10 ALA 1.07427S10 CYS 1.07427S10 ASP 1.07427S10 GLU 1.07427S10 PHE 1.07427S10 GLY 1.07427S10 HIS 1.07427S10 ILE 1.07427S10 LYS -3.32534S10 LEU -2.08882S10 MET -4.85527S10 ASN 1.07427S10 PRO 1.07427S10 GLN -4.39035S10 ARG 1.07427S10 SER 1.07427S10 THR -2.52858S10 VAL 1.07427S10 TRP 1.07427S10 TYR 1.07427S10 1.07427Sil ALA 0.374352Sil CYS 0.374352Sil ASP 0.374352Sil GLU 0.374352Sil PHE 0.374352Sil GLY 0.374352Sil HIS 0.374352Sil ILE 0.374352Sil LYS 0.374352Sil LEU 0.374352Sil MET 0.374352Sil ASN 0.374352Sil PRO 0.374352Sil GLN -7.48704Sil ARG 0.374352Sil SER 0.374352Sil THR 0.374352Sil VAL 0.374352Sil TRP 0.374352Sil TYR 0.374352Sil 0.374352S12 ALA -3.71744S12 CYS 1.18785S12 ASP -3.04852S12 GLU 1.18785S12 PHE 1.18785S12 GLY 1.18785S12 HIS -3.14202S12 ILE 1.18785S12 LYS 1.18785S12 LEU 1.18785S12 MET 1.18785S12 ASN 0.188663S12 PRO 1.18785S12 GLN 1.18785S12 ARG 1.18785S12 SER -4.66725S12 THR -3.43113S12 VAL 1.18785S12 TRP 1.18785S12 TYR 1.18785S12 1.18785S13 ALA 0.961887S13 CYS 0.961887S13 ASP -2.90209S13 GLU 0.961887S13 PHE 0.961887S13 GLY 0.961887S13 HIS 0.961887S13 ILE 0.961887S13 LYS 0.961887S13 LEU -5.64273S13 MET 0.961887S13 ASN 0.961887S13 PRO -2.85926S13 GLN 0.961887S13 ARG 0.961887S13 SER 0.961887S13 THR -2.84373S13 VAL 0.961887S13 TRP 0.961887S13 TYR -1.14238S13 - 0.961887S14 ALA 1.46715S14 CYS 1.46715S14 ASP 1.46715S14 GLU 1.46715S14 PHE -4.01509S14 GLY 1.46715S14 HIS -2.15429S14 ILE 1.46715S14 LYS -2.71889S14 LEU 1.46715S14 MET 1.46715S14 ASN 1.46715S14 PRO 1.46715S14 GLN -3.74874S14 ARG 0.696693S14 SER -1.92429S14 THR -2.38304S14 VAL 1.46715S14 TRP 1.46715S14 TYR -2.82528S14 - 1.46715S15 ALA 0.781994S15 CYS 0.781994S15 ASP 0.781994S15 GLU 0.781994S15 PHE -3.11058S15 GLY 0.781994S15 HIS -5.20635S15 ILE 0.781994S15 LYS 0.781994S15 LEU 0.781994S15 MET 0.781994S15 ASN 0.781994S15 PRO 0.781994S15 GLN 0.781994S15 ARG 0.781994S15 SER 0.781994S15 THR -5.75897S15 VAL 0.781994S15 TRP 0.781994S15 TYR 0.781994S15 - 0.781994S16 ALA 0.733142S16 CYS 0.733142S16 ASP 0.733142S16 GLU 0.733142S16 PHE -0.362268S16 GLY 0.733142S16 HIS 0.733142S16 ILE 0.733142S16 LYS 0.733142S16 LEU -1.57201S16 MET 0.733142S16 ASN 0.733142S16 PRO 0.733142S16 GLN 0.733142S16 ARG 0.733142S16 SER 0.733142S16 THR 0.733142S16 VAL 0.733142S16 TRP -4.3517S16 TYR -6.17744S16 - 0.733142S17 ALA 0.951334S17 CYS 0.951334S17 ASP -4.28382S17 GLU 0.951334S17 PHE -5.15747S17 GLY 0.951334S17 HIS 0.951334S17 ILE 0.951334S17 LYS -2.8882S17 LEU 0.951334S17 MET 0.951334S17 ASN 0.951334S17 PRO 0.951334S17 GLN 0.951334S17 ARG 0.951334S17 SER 0.951334S17 THR 0.951334S17 VAL 0.951334S17 TRP 0.951334S17 TYR -3.84317S17 - 0.951334S18 ALA -3.26733S18 CYS 1.29892S18 ASP 1.29892S18 GLU 1.29892S18 PHE 1.29892S18 GLY 1.29892S18 HIS 1.29892S18 ILE -2.86698S18 LYS 1.29892S18 LEU 1.29892S18 MET 1.29892S18 ASN -3.34842S18 PRO 1.29892S18 GLN -2.25519S18 ARG 1.29892S18 SER -3.46368S18 THR -4.28218S18 VAL 1.29892S18 TRP 1.29892S18 TYR 1.29892S18 - 1.29892S19 ALA 0.696632S19 CYS 0.696632S19 ASP 0.696632S19 GLU 0.696632S19 PHE -6.41695S19 GLY 0.696632S19 HIS 0.696632S19 ILE 0.696632S19 LYS 0.696632S19 LEU 0.696632S19 MET 0.696632S19 ASN 0.696632S19 PRO 0.696632S19 GLN -2.69291S19 ARG 0.696632S19 SER 0.696632S19 THR 0.696632S19 VAL 0.696632S19 TRP -3.42952S19 TYR 0.696632S19 - 0.696632S20 ALA 0.975145S20 CYS 0.975145S20 ASP 0.975145S20 GLU -0.122216S20 PHE 0.975145S20 GLY 0.975145S20 HIS 0.975145S20 ILE 0.975145S20 LYS -2.50824S20 LEU 0.975145S20 MET 0.975145S20 ASN 0.975145S20 PRO 0.975145S20 GLN 0.975145S20 ARG -5.63992S20 SER -4.5873S20 THR -2.74464S20 VAL 0.975145S20 TRP 0.975145S20 TYR 0.975145S20 - 0.975145S21 ALA 0.620787S21 CYS 0.620787S21 ASP -6.82068S21 GLU 0.620787S21 PHE 0.620787S21 GLY 0.620787S21 HIS 0.620787S21 ILE 0.620787S21 LYS 0.620787S21 LEU 0.620787S21 MET 0.620787S21 ASN 0.620787S21 PRO 0.620787S21 GLN 0.620787S21 ARG 0.620787S21 SER -3.83895S21 THR 0.620787S21 VAL 0.620787S21 TRP -0.514539S21 TYR 0.620787S21 - 0.620787S22 ALA 0.771591S22 CYS 0.771591S22 ASP 0.771591S22 GLU 0.771591S22 PHE -4.81622S22 GLY 0.771591S22 HIS 0.771591S22 ILE 0.771591S22 LYS 0.771591S22 LEU 0.771591S22 MET 0.771591S22 ASN 0.771591S22 PRO 0.771591S22 GLN 0.771591S22 ARG 0.771591S22 SER 0.771591S22 THR -3.23609S22 VAL 0.771591S22 TRP 0.771591S22 TYR -5.83634S22 - 0.771591S23 ALA 0.737721S23 CYS 0.737721S23 ASP 0.737721S23 GLU 0.737721S23 PHE 0.737721S23 GLY 0.737721S23 HIS 0.737721S23 ILE -3.34383S23 LYS 0.737721S23 LEU 0.737721S23 MET 0.737721S23 ASN 0.737721S23 PRO 0.737721S23 GLN 0.737721S23 ARG 0.737721S23 SER -6.35944S23 THR -3.5757S23 VAL 0.737721S23 TRP 0.737721S23 TYR 0.737721S23 - 0.737721S24 ALA 0.996126S24 CYS 0.996126S24 ASP -3.77447S24 GLU 0.996126S24 PHE 0.996126S24 GLY -3.88926S24 HIS 0.996126S24 ILE 0.996126S24 LYS 0.996126S24 LEU 0.996126S24 MET 0.996126S24 ASN 0.996126S24 PRO 0.996126S24 GLN 0.996126S24 ARG 0.996126S24 SER -5.00007S24 THR -4.27035S24 VAL 0.996126S24 TRP 0.996126S24 TYR 0.996126S24 - 0.996126S25 ALA 1.16098S25 CYS 1.16098S25 ASP 1.16098S25 GLU 1.16098S25 PHE -2.5124S25 GLY 1.16098S25 HIS 1.16098S25 ILE 1.16098S25 LYS -3.35433S25 LEU 1.16098S25 MET 1.16098S25 ASN -4.92245S25 PRO 1.16098S25 GLN 1.16098S25 ARG 1.16098S25 SER -2.75822S25 THR -4.27873S25 VAL 1.16098S25 TRP 1.16098S25 TYR 0.411442S25 - 1.16098S26 ALA 1.14752S26 CYS 1.14752S26 ASP 1.14752S26 GLU 1.14752S26 PHE 1.14752S26 GLY 1.14752S26 HIS 1.14752S26 ILE -3.49337S26 LYS -4.86848S26 LEU 1.14752S26 MET 1.14752S26 ASN 1.14752S26 PRO -2.4624S26 GLN 1.14752S26 ARG 1.14752S26 SER 1.14752S26 THR -3.87533S26 VAL 1.14752S26 TRP 1.14752S26 TYR -3.6608S26 - 1.14752S27 ALA -5.95322S27 CYS 0.761597S27 ASP 0.761597S27 GLU 0.761597S27 PHE 0.761597S27 GLY 0.761597S27 HIS 0.761597S27 ILE 0.761597S27 LYS 0.761597S27 LEU 0.761597S27 MET 0.761597S27 ASN 0.761597S27 PRO 0.761597S27 GLN 0.761597S27 ARG -3.28895S27 SER 0.761597S27 THR -4.46656S27 VAL 0.761597S27 TRP 0.761597S27 TYR 0.761597S27 - 0.761597S28 ALA 1.64865S28 CYS 1.64865S28 ASP 1.64865S28 GLU -1.71887S28 PHE 1.64865S28 GLY -1.02037S28 HIS -1.61583S28 ILE -1.59768S28 LYS -2.93014S28 LEU 0.790686S28 MET -0.97651S28 ASN 1.64865S28 PRO 1.64865S28 GLN 1.64865S28 ARG -3.9409S28 SER 1.64865S28 THR 1.64865S28 VAL 0.375285S28 TRP -2.85368S28 TYR -0.998431S28 - 1.64865S29 ALA 0.374352S29 CYS 0.374352S29 ASP -7.48703S29 GLU 0.374352S29 PHE 0.374352S29 GLY 0.374352S29 HIS 0.374352S29 ILE 0.374352S29 LYS 0.374352S29 LEU 0.374352S29 MET 0.374352S29 ASN 0.374352S29 PRO 0.374352S29 GLN 0.374352S29 ARG 0.374352S29 SER 0.374352S29 THR 0.374352S29 VAL 0.374352S29 TRP 0.374352S29 TYR 0.374352S29 - 0.374352S30 ALA -4.50574S30 CYS 0.590571S30 ASP 0.590571S30 GLU 0.590571S30 PHE 0.590571S30 GLY 0.590571S30 HIS 0.590571S30 ILE 0.590571S30 LYS 0.590571S30 LEU 0.590571S30 MET 0.590571S30 ASN 0.590571S30 PRO 0.590571S30 GLN 0.590571S30 ARG -6.7151S30 SER 0.590571S30 THR 0.590571S30 VAL 0.590571S30 TRP 0.590571S30 TYR 0.590571S30 - 0.590571S31 ALA -1.20828S31 CYS 1.90403S31 ASP 1.90403S31 GLU -1.40079S31 PHE -1.33822S31 GLY -1.3408S31 HIS 1.20301S31 ILE -2.03803S31 LYS -0.800861S31 LEU -3.17664S31 MET 1.90403S31 ASN 1.90403S31 PRO 1.90403S31 GLN 1.90403S31 ARG -1.37676S31 SER -1.7164S31 THR 1.90403S31 VAL 1.90403S31 TRP -2.47325S31 TYR -1.46927S31 - 1.90403S32 ALA 0.940523S32 CYS 0.940523S32 ASP 0.940523S32 GLU 0.940523S32 PHE -2.9074S32 GLY -2.84362S32 HIS 0.940523S32 ILE 0.940523S32 LYS 0.940523S32 LEU 0.940523S32 MET 0.940523S32 ASN 0.940523S32 PRO 0.940523S32 GLN 0.940523S32 ARG 0.940523S32 SER -5.06444S32 THR -5.17343S32 VAL 0.940523S32 TRP 0.940523S32 TYR 0.940523S32 0.940523S33 ALA -3.11711S33 CYS 1.49682S33 ASP 0.748273S33 GLU -1.98311S33 PHE 1.49682S33 GLY 1.49682S33 HIS 1.49682S33 ILE -4.0465S33 LYS -0.270053S33 LEU 1.49682S33 MET 1.49682S33 ASN -0.246105S33 PRO 1.49682S33 GLN 0.602298S33 ARG -0.135242S33 SER -3.7326S33 THR -1.81728S33 VAL -0.970764S33 TRP 1.49682S33 TYR 1.49682S33 1.49682S34 ALA -3.68035S34 CYS 1.05171S34 ASP 1.05171S34 GLU 1.05171S34 PHE 1.05171S34 GLY 1.05171S34 HIS 1.05171S34 ILE 1.05171S34 LYS -0.138403S34 LEU 1.05171S34 MET 1.05171S34 ASN 1.05171S34 PRO 1.05171S34 GLN 1.05171S34 ARG -4.22279S34 SER -4.99245S34 THR -3.79337S34 VAL 1.05171S34 TRP 1.05171S34 TYR 1.05171S34 1.05171S35 ALA -3.46819S35 CYS 1.40614S35 ASP 1.40614S35 GLU -3.56246S35 PHE -1.33106S35 GLY 1.40614S35 HIS 1.40614S35 ILE -3.59998S35 LYS 1.40614S35 LEU -3.13508S35 MET 1.40614S35 ASN 1.40614S35 PRO 1.40614S35 GLN 1.40614S35 ARG 1.40614S35 SER 1.40614S35 THR -1.1115S35 VAL -3.47768S35 TRP 1.40614S35 TYR 1.40614S35 1.40614S36 ALA -2.06924S36 CYS 1.17995S36 ASP -2.50762S36 GLU 1.17995S36 PHE 1.17995S36 GLY 1.17995S36 HIS 1.17995S36 ILE 1.17995S36 LYS -2.9499S36 LEU 1.17995S36 MET 1.17995S36 ASN 1.17995S36 PRO 1.17995S36 GLN 1.17995S36 ARG -5.25862S36 SER -2.43951S36 THR 1.17995S36 VAL 1.17995S36 TRP -2.47433S36 TYR 1.17995S36 1.17995S37 ALA 1.55809S37 CYS 1.55809S37 ASP 1.55809S37 GLU -2.46647S37 PHE 1.55809S37 GLY 1.55809S37 HIS 1.55809S37 ILE -2.74202S37 LYS -0.87747S37 LEU 1.55809S37 MET 1.55809S37 ASN 0.560965S37 PRO -1.90277S37 GLN 1.55809S37 ARG -3.18005S37 SER 0.3709S37 THR 0.169053S37 VAL -3.4419S37 TRP 1.55809S37 TYR -3.62927S37 1.55809S38 ALA -2.50337S38 CYS 1.53418S38 ASP 1.53418S38 GLU 1.53418S38 PHE -2.45495S38 GLY 1.53418S38 HIS 1.53418S38 ILE -2.54748S38 LYS 1.53418S38 LEU 1.53418S38 MET 1.53418S38 ASN 1.53418S38 PRO 1.53418S38 GLN 1.53418S38 ARG -2.04271S38 SER -1.18619S38 THR 1.53418S38 VAL -4.10348S38 TRP -1.83638S38 TYR -3.26974S38 1.53418S39 ALA 1.32683S39 CYS 1.32683S39 ASP 1.32683S39 GLU 1.32683S39 PHE -3.5779S39 GLY 1.32683S39 HIS 1.32683S39 ILE 1.32683S39 LYS 1.32683S39 LEU -3.53985S39 MET -3.0458S39 ASN -4.11409S39 PRO -2.28858S39 GLN 1.32683S39 ARG 1.32683S39 SER 1.32683S39 THR 1.32683S39 VAL 1.32683S39 TRP 1.32683S39 TYR -3.33627S39 1.32683S40 ALA -4.42109S40 CYS 0.911932S40 ASP -5.4143S40 GLU 0.911932S40 PHE 0.911932S40 GLY 0.911932S40 HIS 0.911932S40 ILE 0.911932S40 LYS 0.911932S40 LEU 0.911932S40 MET 0.911932S40 ASN -2.99386S40 PRO 0.911932S40 GLN 0.911932S40 ARG -2.6736S40 SER 0.911932S40 THR 0.911932S40 VAL 0.911932S40 TRP 0.911932S40 TYR 0.911932540 - 0.911932541 ALA 0.371475S41 CYS 1.25366S41 ASP -2.30521S41 GLU 1.25366S41 PHE 1.25366S41 GLY 1.25366S41 HIS -2.3859S41 ILE -4.07155S41 LYS 1.25366S41 LEU -2.30675S41 MET 1.25366S41 ASN 1.25366S41 PRO 1.25366S41 GLN 1.25366S41 ARG 1.25366S41 SER -2.3004S41 THR 1.25366S41 VAL -4.55291S41 TRP 1.253 66 S41 TYR 1.253 66 S41 - 1.253 66 SO SI ALA GLU 1.88968e-05 SO SI ALA LEU 0.000311545 SO SI ALA SER 2.06957e-05 SO SI ALA VAL 0.000206061 SO SI CYS GLU 1.88968e-05 SO SI CYS LEU 0.000311545 SO SI CYS SER 2.06957e-05 SO SI CYS VAL 0.000206061 SO SI ASP GLU 1.88968e-05 SO SI ASP LEU 0.000311545 SO SI ASP SER 2.06957e-05 SO SI ASP VAL 0.000206061 SO SI GLU ALA 0.0001171 SO SI GLU CYS 0.0001171 SO SI GLU ASP 0.0001171 SO SI GLU GLU 0.00758128 SO SI GLU PHE 0.0001171 SO SI GLU GLY 0.0001171 SO SI GLU HIS 0.0001171 SO SI GLU ILE 0.0001171 SO SI GLU LYS 0.0001171 SO SI GLU LEU 0.0615903 SO SI GLU MET 0.0001171 SO SI GLU ASN 0.0001171 SO SI GLU PRO 0.0001171 SO SI GLU GLN 0.0001171 SO SI GLU ARG 0.0001171 SO SI GLU SER 0.00743999S41 THR LEU 2.19518e-05 S41 THR SER 3.49475e-05 S41 THR VAL 0.000295087 S41 VAL ALA 9.24962e-07 S41 VAL ASP 2.05039e-05 S41 VAL HIS 1.50841e-05 S41 VAL ILE 0.000168699 S41 VAL LEU 2.19518e-05 S41 VAL SER 3.49475e-05 S41 VAL VAL 0.000295087 S41 TRP ALA 9.24962e-07 S41 TRP ASP 2.05039e-05 S41 TRP HIS 1.50841e-05 S41 TRP ILE 0.000168699 S41 TRP LEU 2.19518e-05 S41 TRP SER 3.49475e-05 S41 TRP VAL 0.000295087 S41 TYR ALA 9.24962e-07 S41 TYR ASP 2.05039e-05 S41 TYR HIS 1.50841e-05 S41 TYR ILE 0.000168699 S41 TYR LEU 2.19518e-05 S41 TYR SER 3.49475e-05 S41 TYR VAL 0.000295087 S41 - ALA 9.24962e-07 S41 - ASP 2.05039e-05 S41 - HIS 1.50841e-05 S41 - ILE 0.000168699 S41 - LEU 2.19518e-05 S41 - SER 3.49475e-05 S41 - VAL 0.000295087 S2 ALA ALA 1.03615e-05 S2 ALA ASP 7.54619e-05 S2 ALA PHE 2.74683e-05 S2 ALA LEU 0.000330065 S2 ALA GLN 1.80009e-05 S2 ALA ARG 8.15538e-05 S2 ALA SER 1.25566e-05 S2 ALA VAL 2.05353e-05 S2 CYS ALA 1.03615e-05 S2 CYS ASP 7.54619e-05 S2 CYS PHE 2.74683e-05 S2 CYS LEU 0.000330065 S2 CYS GLN 1.80009e-05 S2 CYS ARG 8.15538e-05 S2 CYS SER 1.25566e-05 S2 CYS VAL 2.05353e-05 S2 ASP ALA 1.03615e-05 S2 ASP ASP 7.54619e-05 S2 ASP PHE 2.74683e-05 S2 ASP LEU 0.000330065 S2 ASP GLN 1.80009e-05 S2 ASP ARG 8.15538e-05 S2 ASP SER 1.25566e-05 S2 ASP VAL 2.05353e-05 S2 GLU ALA 0.00070969 S2 GLU CYS 2 ,28669e-05S41 SER TRP 3.12437e-05 S41 SER TYR 3.12437e-05 S41 SER - 3.12437e-05 S41 THR ALA 9.61713e-07 S41 THR ASP 2.4559e-05 S41 THR HIS 1.63033e-05 S41 THR ILE 0.00013502 S41 THR LEU 1.9545e-05 S41 THR SER 2.6156e-05 S41 THR VAL 0.000353459 S41 VAL ALA 0.000655793 S41 VAL CYS 0.000205952 S41 VAL ASP 0.0109811 S41 VAL GLU 0.000205952 S41 VAL PHE 0.000205952 S41 VAL GLY 0.000205952 S41 VAL HIS 0.00819723 S41 VAL ILE -0.0402067 S41 VAL LYS 0.000205952 S41 VAL LEU 0.0109351 S41 VAL MET 0.000205952 S41 VAL ASN 0.000205952 S41 VAL PRO 0.000205952 S41 VAL GLN 0.000205952 S41 VAL ARG 0.000205952 S41 VAL SER -0.0295014 S41 VAL THR 0.000205952 S41 VAL VAL 0.0332202 S41 VAL TRP 0.000205952 S41 VAL TYR 0.000205952 S41 VAL - 0.000205952 S41 TRP ALA 9.61713e-07 S41 TRP ASP 2.4559e-05 S41 TRP HIS 1.63033e-05 S41 TRP ILE 0.00013502 S41 TRP LEU 1.9545e-05 S41 TRP SER 2.6156e-05 S41 TRP VAL 0.000353459 S41 TYR ALA 9.61713e-07 S41 TYR ASP 2.4559e-05 S41 TYR HIS 1.63033e-05 S41 TYR ILE 0.00013502 S41 TYR LEU 1.9545e-05 S41 TYR SER 2.6156e-05 S41 TYR VAL 0.000353459 S41 - ALA 9.61713e-07 S41 - ASP 2.4559e-05 S41 - HIS 1.63033e-05 S41 - ILE 0.00013502 S41 - LEU 1.9545e-05 S41 - SER 2.6156e-05 S41 - VAL 0.000353459 S3 ALA ALA 6.59033e-06 S3 ALA CYS 6.59033e-06 S3 ALA ASP 6.59033e-06 S3 ALA GLU 6.59033e-06 S3 ALA PHE 6.59033e-06S41 TRP LEU 4.21964e-05 S41 TRP SER 4.20388e-05 S41 TRP VAL 0.000540022 S41 TYR ALA 1.68983e-06 S41 TYR ASP 3.04494e-05 S41 TYR HIS 2.35041e-05 S41 TYR ILE 0.000222142 S41 TYR LEU 4.21964e-05 S41 TYR SER 4.20388e-05 S41 TYR VAL 0.000540022 S41 - ALA 1.68983e-06 S41 - ASP 3.04494e-05 S41 - HIS 2.35041e-05 S41 - ILE 0.000222142 S41 - LEU 4.21964e-05 S41 - SER 4.20388e-05 S41 - VAL 0.000540022 S4 ALA GLU 5.01377e-05 S4 ALA HIS 9.26495e-06 S4 ALA ILE 4.86085e-05 S4 ALA LYS 6.38285e-07 S4 ALA LEU 1.44678e-05 S4 ALA GLN 4.24909e-06 S4 ALA ARG 1.40732e-05 S4 ALA SER 4.1197e-05 S4 ALA THR 2.8341e-05 S4 ALA VAL 0.000118406 S4 ALA TYR 6.7449e-06 S4 CYS GLU 5.01377e-05 S4 CYS HIS 9.26495e-06 S4 CYS ILE 4.86085e-05 S4 CYS LYS 6.38285e-07 S4 CYS LEU 1.44678e-05 S4 CYS GLN 4.24909e-06 S4 CYS ARG 1.40732e-05 S4 CYS SER 4.1197e-05 S4 CYS THR 2.8341e-05 S4 CYS VAL 0.000118406 S4 CYS TYR 6.7449e-06 S4 ASP GLU 5.01377e-05 S4 ASP HIS 9.26495e-06 S4 ASP ILE 4.86085e-05 S4 ASP LYS 6.38285e-07 S4 ASP LEU 1.44678e-05 S4 ASP GLN 4.24909e-06 S4 ASP ARG 1.40732e-05 S4 ASP SER 4.1197e-05 S4 ASP THR 2.8341e-05 S4 ASP VAL 0.000118406 S4 ASP TYR 6.7449e-06 S4 GLU GLU 5.01377e-05 S4 GLU HIS 9.26495e-06 S4 GLU ILE 4.86085e-05 S4 GLU LYS 6.38285e-07 S4 GLU LEU 1.44678e-05 S4 GLU GLN 4.24909e-06 S4 GLU ARG 1.40732e-05S41 SER SER -0.00268126 S41 SER THR 0.000768007 S41 SER VAL 0.00647686 S41 SER TRP 0.000768007 S41 SER TYR 0.000768007 S41 SER - 0.000768007 S41 THR ALA 7.54409e-05 S41 THR CYS 1.59096e-05 S41 THR ASP 0.0012413 S41 THR GLU 1.59096e-05 S41 THR PHE 1.59096e-05 S41 THR GLY 1.59096e-05 S41 THR HIS 0.00121538 S41 THR ILE 0.00629243 S41 THR LYS 1.59096e-05 S41 THR LEU 0.00123868 S41 THR MET 1.59096e-05 S41 THR ASN 1.59096e-05 S41 THR PRO 1.59096e-05 S41 THR GLN 1.59096e-05 S41 THR ARG 1.59096e-05 S41 THR SER 0.0010061 S41 THR THR 1.59096e-05 S41 THR VAL -0.0133091 S41 THR TRP 1.59096e-05 S41 THR TYR 1.59096e-05 S41 THR - 1.59096e-05 S41 VAL ALA 7.48777 e- 07 S41 VAL ASP 1.37407e-05 S41 VAL HIS 1.19263e-05 S41 VAL ILE 7.68526e-05 S41 VAL LEU 1.32211e-05 S41 VAL SER 1.26327e-05 S41 VAL VAL 0.000207007 S41 TRP ALA 7.48777 e- 07 S41 TRP ASP 1.37407e-05 S41 TRP HIS 1.19263e-05 S41 TRP ILE 7.68526e-05 S41 TRP LEU 1.32211e-05 S41 TRP SER 1.26327e-05 S41 TRP VAL 0.000207007 S41 TYR ALA 7.48777 e- 07 S41 TYR ASP 1.37407e-05 S41 TYR HIS 1.19263e-05 S41 TYR ILE 7.68526e-05 S41 TYR LEU 1.32211e-05 S41 TYR SER 1.26327e-05 S41 TYR VAL 0.000207007 S41 - ALA 7.48777e-07 S41 - ASP 1.37407e-05 S41 - HIS 1.19263e-05 S41 - ILE 7.68526e-05 S41 - LEU 1.32211e-05 S41 - SER 1.26327e-05 S41 - VAL 0.000207007 S5 ALA HIS 9.0571e-05 S5 ALA LYS 0.000380982S4 S30 SER ASP 4.70908e-05S4 S30 SER GLU 4.70908e-05S4 S30 SER PHE 4.70908e-05S4 S30 SER GLY 4.70908e-05S4 S30 SER HIS 4.70908e-05S4 S30 SER ILE 4.70908e-05S4 S30 SER LYS 4.70908e-05S4 S30 SER LEU 4.70908e-05S4 S30 SER MET 4.70908e-05S4 S30 SER ASN 4.70908e-05S4 S30 SER PRO 4.70908e-05S4 S30 SER GLN 4.70908e-05S4 S30 SER ARG -0.0165873S4 S30 SER SER 4.70908e-05S4 S30 SER THR 4.70908e-05S4 S30 SER VAL 4.70908e-05S4 S30 SER TRP 4.70908e-05S4 S30 SER TYR 4.70908e-05S4 S30 SER - 4.70908e-05S4 S30 THR ALA 0.00871227S4 S30 THR CYS 3.0944e-05S4 S30 THR ASP 3.0944e-05S4 S30 THR GLU 3.0944e-05S4 S30 THR PHE 3.0944e-05S4 S30 THR GLY 3.0944e-05S4 S30 THR HIS 3.0944e-05S4 S30 THR ILE 3.0944e-05S4 S30 THR LYS 3.0944e-05S4 S30 THR LEU 3.0944e-05S4 S30 THR MET 3.0944e-05S4 S30 THR ASN 3.0944e-05S4 S30 THR PRO 3.0944e-05S4 S30 THR GLN 3.0944e-05S4 S30 THR ARG -0.0107502S4 S30 THR SER 3.0944e-05S4 S30 THR THR 3.0944e-05S4 S30 THR VAL 3.0944e-05S4 S30 THR TRP 3.0944e-05S4 S30 THR TYR 3.0944e-05S4 S30 THR - 3.0944e-05S4 S30 VAL ALA 0.0276675S4 S30 VAL CYS 9.72146e-05S4 S30 VAL ASP 9.72146e-05S4 S30 VAL GLU 9.72146e-05S4 S30 VAL PHE 9.72146e-05S4 S30 VAL GLY 9.72146e-05S4 S30 VAL HIS 9.72146e-05S4 S30 VAL ILE 9.72146e-05S4 S30 VAL LYS 9.72146e-05S4 S30 VAL LEU 9.72146e-05S4 S30 VAL MET 9.72146e-05S4 S30 VAL ASN 9.72146e-05S4 S30 VAL PRO 9.72146e-05S4 S30 VAL GLN 9.72146e-05S4 S30 VAL ARG -0.0316446S4 S30 VAL SER 9.72146e-05S4 S30 VAL THR 9.72146e-05-771 -S4 S31 HIS PHE 0.00132664S4 S31 HIS GLY 0.00123175S4 S31 HIS HIS 9.534856-05S4 S31 HIS ILE 0.0033664S4 S31 HIS LYS 0.00112952S4 S31 HIS LEU -0.0149112S4 S31 HIS MET 2.738486-05S4 S31 HIS ASN 2.738486-05S4 S31 HIS PRO 2.738486-05S4 S31 HIS GLN 2.738486-05S4 S31 HIS ARG 0.00121444S4 S31 HIS SER 0.00176301S4 S31 HIS THR 2.738486-05S4 S31 HIS VAL 2.738486-05S4 S31 HIS TRP -0.00101299S4 S31 HIS TYR 0.00149637S4 S31 HIS - 2.738486-05S4 S31 ILE ALA 0.00911888S4 S31 ILE CYS 0.000205937S4 S31 ILE ASP 0.000205937S4 S31 ILE GLU -0.000186856S4 S31 ILE PHE 0.00692141S4 S31 ILE GLY 0.00666862S4 S31 ILE HIS -0.00217018S4 S31 ILE ILE -0.00525795S4 S31 ILE LYS 0.00585871S4 S31 ILE LEU 0.0328023S4 S31 ILE MET 0.000205937S4 S31 ILE ASN 0.000205937S4 S31 ILE PRO 0.000205937S4 S31 ILE GLN 0.000205937S4 S31 ILE ARG 0.00777107S4 S31 ILE SER -0.0565054S4 S31 ILE THR 0.000205937S4 S31 ILE VAL 0.000205937S4 S31 ILE TRP -0.0171504S4 S31 ILE TYR 0.00864777S4 S31 ILE - 0.000205937S4 S31 LYS ALA 0.00014029S4 S31 LYS CYS 1.900766-06S4 S31 LYS ASP 1.90076e-06S4 S31 LYS GLU 0.000178569S4 S31 LYS PHE 0.000128983S4 S31 LYS GLY 0.000110397S4 S31 LYS HIS 8.502236-06S4 S31 LYS ILE 0.00031993S4 S31 LYS LYS 0.000107059S4 S31 LYS LEU -0.00123869S4 S31 LYS MET 1.90076e-06S4 S31 LYS ASN 1.90076e-06S4 S31 LYS PRO 1.900766-06S4 S31 LYS GLN 1.900766-06S4 S31 LYS ARG 0.000120545S4 S31 LYS SER 0.000172068S4 S31 LYS THR 1.90076e-06S4 S31 LYS VAL 1.90076e-06S4 S31 LYS TRP 0.000511056-774-S4 S31 SER GLN 0.000168868S4 S31 SER ARG 0.00683078S4 S31 SER SER 0.0102897S4 S31 SER THR 0.000168868S4 S31 SER VAL 0.000168868S4 S31 SER TRP -0.0198181S4 S31 SER TYR 0.000192032S4 S31 SER - 0.000168868S4 S31 THR ALA 0.00455955S4 S31 THR CYS 0.000112653S4 S31 THR ASP 0.000112653S4 S31 THR GLU 0.00559298S4 S31 THR PHE 0.00584095S4 S31 THR GLY 0.0066781S4 S31 THR HIS 0.000361301S4 S31 THR ILE 0.0115048S4 S31 THR LYS 0.00307416S4 S31 THR LEU -0.0240456S4 S31 THR MET 0.000112653S4 S31 THR ASN 0.000112653S4 S31 THR PRO 0.000112653S4 S31 THR GLN 0.000112653S4 S31 THR ARG -0.0451863S4 S31 THR SER 0.00709354S4 S31 THR THR 0.000112653S4 S31 THR VAL 0.000112653S4 S31 THR TRP 0.016343S4 S31 THR TYR 0.00572003S4 S31 THR - 0.000112653S4 S31 VAL ALA 0.012901S4 S31 VAL CYS 0.000361401S4 S31 VAL ASP 0.000361401S4 S31 VAL GLU 0.0204687S4 S31 VAL PHE 0.0117687S4 S31 VAL GLY -0.0330917S4 S31 VAL HIS 0.000962441S4 S31 VAL ILE -0.0609191S4 S31 VAL LYS 0.0118355S4 S31 VAL LEU 0.00650356S4 S31 VAL MET 0.000361401S4 S31 VAL ASN 0.000361401S4 S31 VAL PRO 0.000361401S4 S31 VAL GLN 0.000361401S4 S31 VAL ARG 0.014594S4 S31 VAL SER 0.0175447S4 S31 VAL THR 0.000361401S4 S31 VAL VAL 0.000361401S4 S31 VAL TRP 0.0218727S4 S31 VAL TYR -0.0298235S4 S31 VAL - 0.000361401S4 S31 TRP ALA 4.767076-05S4 S31 TRP GLU 6.32856-05S4 S31 TRP PHE 4.484416-05S4 S31 TRP GLY 3.637496-05S4 S31 TRP HIS 2.463036-06S4 S31 TRP ILE 0.000104608S4 S31 TRP LYS 3.570566-05-777-S41 VAL PRO 4.32829e-05 S41 VAL GLN 4.32829e-05 S41 VAL ARG 4.32829e-05 S41 VAL SER 0.00199443 S41 VAL THR 4.32829e-05 S41 VAL VAL 0.0323841 S41 VAL TRP 4.32829e-05 S41 VAL TYR 4.32829e-05 S41 VAL - 4.32829e-05 S41 TRP ALA 1.60515e-06 S41 TRP ASP 4.28314e-05 S41 TRP HIS 2.45582e-05 S41 TRP ILE 0.000182265 S41 TRP LEU 2.36206e-05 S41 TRP SER 2.20062e-05 S41 TRP VAL 0.000387122 S41 TYR ALA -0.00144482 S41 TYR CYS 0.0003774 S41 TYR ASP 0.0242772 S41 TYR GLU 0.0003774 S41 TYR PHE 0.0003774 S41 TYR GLY 0.0003774 S41 TYR HIS -0.027154 S41 TYR ILE 0.0295294 S41 TYR LYS 0.0003774 S41 TYR LEU -0.000578575 S41 TYR MET 0.0003774 S41 TYR ASN 0.0003774 S41 TYR PRO 0.0003774 S41 TYR GLN 0.0003774 S41 TYR ARG 0.0003774 S41 TYR SER -0.0218204 S41 TYR THR 0.0003774 S41 TYR VAL -0.0112013 S41 TYR TRP 0.0003774 S41 TYR TYR 0.0003774 S41 TYR - 0.0003774 S41 - ALA 1.60515e-06 S41 - ASP 4.28314e-05 S41 - HIS 2.45582e-05 S41 - ILE 0.000182265 S41 - LEU 2.36206e-05 S41 - SER 2.20062e-05 S41 - VAL 0.000387122 S7 ALA ALA 4.00114e-05 S7 ALA CYS 3.93239e-05 S7 ALA GLU 1.93586e-05 S7 ALA LYS 8.4318e-05 S7 ALA LEU 0.000279552 S7 ALA ARG 9.73859e-05 S7 ALA THR 2.04181e-05 S7 CYS ALA 0.00434345 S7 CYS CYS -0.0522341 S7 CYS ASP 4.76732e-05 S7 CYS GLU 0.00226503 S7 CYS PHE 4.76732e-05 S7 CYS GLY 4.76732e-05S41 SER LEU 0.00385692 S41 SER MET 8.30199e-05 S41 SER ASN 8.30199e-05 S41 SER PRO 8.30199e-05 S41 SER GLN 8.30199e-05 S41 SER ARG 8.30199e-05 S41 SER SER -0.0457959 S41 SER THR 8.30199e-05 S41 SER VAL 0.0412805 S41 SER TRP 8.30199e-05 S41 SER TYR 8.30199e-05 S41 SER - 8.30199e-05 S41 THR ALA 1.13481e-06 S41 THR ASP 1.96378e-05 S41 THR HIS 1.47935e-05 S41 THR ILE 0.000174286 S41 THR LEU 1.68531e-05 S41 THR SER 4.21229e-05 S41 THR VAL 0.000311539 S41 VAL ALA 1.13481e-06 S41 VAL ASP 1.96378e-05 S41 VAL HIS 1.47935e-05 S41 VAL ILE 0.000174286 S41 VAL LEU 1.68531e-05 S41 VAL SER 4.21229e-05 S41 VAL VAL 0.000311539 S41 TRP ALA 1.13481e-06 S41 TRP ASP 1.96378e-05 S41 TRP HIS 1.47935e-05 S41 TRP ILE 0.000174286 S41 TRP LEU 1.68531e-05 S41 TRP SER 4.21229e-05 S41 TRP VAL 0.000311539 S41 TYR ALA 1.13481e-06 S41 TYR ASP 1.96378e-05 S41 TYR HIS 1.47935e-05 S41 TYR ILE 0.000174286 S41 TYR LEU 1.68531e-05 S41 TYR SER 4.21229e-05 S41 TYR VAL 0.000311539 S41 - ALA 1.13481e-06 S41 - ASP 1.96378e-05 S41 - HIS 1.47935e-05 S41 - ILE 0.000174286 S41 - LEU 1.68531e-05 S41 - SER 4.21229e-05 S41 - VAL 0.000311539 S8 ALA ALA 0.00248326 S8 ALA CYS 5.02184e-05 S8 ALA ASP 5.02184e-05 S8 ALA GLU 5.02184e-05 S8 ALA PHE 5.02184e-05 S8 ALA GLY 0.0158826 S8 ALA HIS 5.02184e-05 S8 ALA ILE 5.02184e-05 S8 ALA LYS 5.02184e-05 S8 ALA LEU 5.02184e-05S7 S22 LEU - 0.00024207S7 S22 MET PHE 0.000206992S7 S22 MET THR 4.76732e-05S7 S22 MET TYR 0.000604169S7 S22 ASN PHE 0.000206992S7 S22 ASN THR 4.76732e-05S7 S22 ASN TYR 0.000604169S7 S22 PRO PHE 0.000206992S7 S22 PRO THR 4.76732e-05S7 S22 PRO TYR 0.000604169S7 S22 GLN PHE 0.000206992S7 S22 GLN THR 4.76732e-05S7 S22 GLN TYR 0.000604169S7 S22 ARG ALA 7.12807e-05S7 S22 ARG CYS 7.12807e-05S7 S22 ARG ASP 7.12807e-05S7 S22 ARG GLU 7.12807e-05S7 S22 ARG PHE -0.00595361S7 S22 ARG GLY 7.12807e-05S7 S22 ARG HIS 7.12807e-05S7 S22 ARG ILE 7.12807e-05S7 S22 ARG LYS 7.12807e-05S7 S22 ARG LEU 7.12807e-05S7 S22 ARG MET 7.12807e-05S7 S22 ARG ASN 7.12807e-05S7 S22 ARG PRO 7.12807e-05S7 S22 ARG GLN 7.12807e-05S7 S22 ARG ARG 7.12807e-05S7 S22 ARG SER 7.12807e-05S7 S22 ARG THR 0.00709367S7 S22 ARG VAL 7.12807e-05S7 S22 ARG TRP 7.12807e-05S7 S22 ARG TYR -0.0044313S7 S22 ARG - 7.12807e-05S7 S22 SER PHE 0.000206992S7 S22 SER THR 4.76732e-05S7 S22 SER TYR 0.000604169S7 S22 THR ALA 2.949e-05S7 S22 THR CYS 2.949e-05S7 S22 THR ASP 2.949e-05S7 S22 THR GLU 2.949e-05S7 S22 THR PHE -0.0438628S7 S22 THR GLY 2.949e-05S7 S22 THR HIS 2.949e-05S7 S22 THR ILE 2.949e-05S7 S22 THR LYS 2.949e-05S7 S22 THR LEU 2.949e-05S7 S22 THR MET 2.949e-05S7 S22 THR ASN 2.949e-05S7 S22 THR PRO 2.949e-05S7 S22 THR GLN 2.949e-05S7 S22 THR ARG 2.949e-05S7 S22 THR SER 2.949e-05S7 S22 THR THR 0.00344317S7 S22 THR VAL 2.949e-05S7 S22 THR TRP 2.949e-05S7 S22 THR TYR 0.0386397- mi -S41 TRP LEU 2.72761e-05 S41 TRP SER 3.47941e-05 S41 TRP VAL 0.000366764 S41 TYR ALA 1.22588e-06 S41 TYR ASP 2.24948e-05 S41 TYR HIS 1.76038e-05 S41 TYR ILE 0.000156502 S41 TYR LEU 2.72761e-05 S41 TYR SER 3.47941e-05 S41 TYR VAL 0.000366764 S41 - ALA 1.22588e-06 S41 - ASP 2.24948e-05 S41 - HIS 1.76038e-05 S41 - ILE 0.000156502 S41 - LEU 2.72761e-05 S41 - SER 3.47941e-05 S41 - VAL 0.000366764 S10 ALA LYS 6.08126e-05 S10 ALA LEU 1.19398e-05 S10 ALA MET 0.000300904 S10 ALA GLN 0.000176064 S10 ALA THR 2.48351e-05 S10 CYS LYS 6.08126e-05 S10 CYS LEU 1.19398e-05 S10 CYS MET 0.000300904 S10 CYS GLN 0.000176064 S10 CYS THR 2.48351e-05 S10 ASP ALA 0.00018849 S10 ASP CYS 0.00018849 S10 ASP ASP 0.00018849 S10 ASP GLU 0.00018849 S10 ASP PHE 0.00018849 S10 ASP GLY 0.00018849 S10 ASP HIS 0.00018849 S10 ASP ILE 0.00018849 S10 ASP LYS -0.0567103 S10 ASP LEU 0.00681297 S10 ASP MET 0.0355068 S10 ASP ASN 0.00018849 S10 ASP PRO 0.00018849 S10 ASP GLN -0.00663106 S10 ASP ARG 0.00018849 S10 ASP SER 0.00018849 S10 ASP THR 0.015233 S10 ASP VAL 0.00018849 S10 ASP TRP 0.00018849 S10 ASP TYR 0.00018849 S10 ASP - 0.00018849 S10 GLU LYS 6.08126e-05 S10 GLU LEU 1.19398e-05 S10 GLU MET 0.000300904 S10 GLU GLN 0.000176064 S10 GLU THR 2.48351e-05 S10 PHE LYS 6.08126e-05 S10 PHE LEU 1.19398e-05 S10 PHE MET 0.000300904 S10 PHE GLN 0.000176064S9 S41 VAL HIS 1 57903e-05S9 S41 VAL ILE 0 000139944S9 S41 VAL LEU 1 83874e-05S9 S41 VAL SER 2 98363e-05S9 S41 VAL VAL 0 000324402S9 S41 TRP ALA 1 03709e-06S9 S41 TRP ASP 4 51576e-05S9 S41 TRP HIS 1 57903e-05S9 S41 TRP ILE 0 000139944S9 S41 TRP LEU 1 83874e-05S9 S41 TRP SER 2 98363e-05S9 S41 TRP VAL 0 000324402S9 S41 TYR ALA 1 03709e-06S9 S41 TYR ASP 4 51576e-05S9 S41 TYR HIS 1 57903e-05S9 S41 TYR ILE 0 000139944S9 S41 TYR LEU 1 83874e-05S9 S41 TYR SER 2 98363e-05S9 S41 TYR VAL 0 000324402S9 S41 ALA 1 03709e-06S9 S41 ASP 4 51576e-05S9 S41 HIS 1 57903e-05S9 S41 ILE 0 000139944S9 S41 LEU 1 83874e-05S9 S41 SER 2 98363e-05S9 S41 VAL 0 000324402S10 Sil ALA GLN 0.000669087S10 Sil CYS GLN 0.000669087S10 Sil ASP GLN 0.000669087S10 Sil GLU GLN 0.000669087S10 Sil PHE GLN 0.000669087S10 Sil GLY GLN 0.000669087S10 Sil HIS GLN 0.000669087S10 Sil ILE GLN 0.000669087S10 Sil LYS ALA 1.96973e-05S10 Sil LYS CYS 1.96973e-05S10 Sil LYS ASP 1.96973e-05S10 Sil LYS GLU 1.96973e-05S10 Sil LYS PHE 1.96973e-05S10 Sil LYS GLY 1.96973e-05S10 Sil LYS HIS 1.96973e-05S10 Sil LYS ILE 1.96973e-05S10 Sil LYS LYS 1.96973e-05S10 Sil LYS LEU 1.96973e-05S10 Sil LYS MET 1.96973e-05S10 Sil LYS ASN 1.96973e-05S10 Sil LYS PRO 1.96973e-05S10 Sil LYS GLN -0.00243929S10 Sil LYS ARG 1.96973e-05S10 Sil LYS SER 1.96973e-05S10 Sil LYS THR 1.96973e-05S10 Sil LYS VAL 1.96973e-05S10 Sil LYS TRP 1.96973e-05S10 Sil LYS TYR 1.96973e-05S10 Sil LYS - 1.96973e-05S10 Sil LEU ALA 5.41977e-06S10 Sil LEU CYS 5.41977e-06S10 Sil LEU ASP 5.41977e-06S10 Sil LEU GLU 5.41977e-06S10 Sil LEU PHE 5.41977e-06S10 Sil LEU GLY 5.41977e-06S10 Sil LEU HIS 5.41977e-06S10 Sil LEU ILE 5.41977e-06S10 Sil LEU LYS 5.41977e-06S10 Sil LEU LEU 5.41977e-06S10 Sil LEU MET 5.41977e-06S10 Sil LEU ASN 5.41977e-06S10 Sil LEU PRO 5.41977e-06S10 Sil LEU GLN -0.00141123S10 Sil LEU ARG 5.41977e-06S10 Sil LEU SER 5.41977e-06S10 Sil LEU THR 5.41977e-06S10 Sil LEU VAL 5.41977e-06S10 Sil LEU TRP 5.41977e-06S10 Sil LEU TYR 5.41977e-06S10 Sil LEU - 5.41977e-06S10 Sil MET ALA 0.000146075S10 Sil MET CYS 0.000146075S10 Sil MET ASP 0.000146075S10 Sil MET GLU 0.000146075S10 Sil MET PHE 0.000146075S10 Sil MET GLY 0.000146075S10 Sil MET HIS 0.000146075S10 Sil MET ILE 0.000146075S10 Sil MET LYS 0.000146075S10 Sil MET LEU 0.000146075S10 Sil MET MET 0.000146075S10 Sil MET ASN 0.000146075S10 Sil MET PRO 0.000146075S10 Sil MET GLN -0.00603735S10 Sil MET ARG 0.000146075S10 Sil MET SER 0.000146075S10 Sil MET THR 0.000146075S10 Sil MET VAL 0.000146075S10 Sil MET TRP 0.000146075S10 Sil MET TYR 0.000146075S10 Sil MET - 0.000146075S10 Sil ASN GLN 0.000669087S10 Sil PRO GLN 0.000669087S10 Sil GLN ALA 4.95542e-05S10 Sil GLN CYS 4.95542e-05S10 Sil GLN ASP 4.95542e-05S10 Sil GLN GLU 4.95542e-05S10 Sil GLN PHE 4.95542e-05S10 Sil GLN GLY 4.95542e-05S10 Sil GLN HIS 4.95542e-05S10 Sil GLN ILE 4.95542e-05S10 Sil GLN LYS 4.95542e-05S10 Sil GLN LEU 4.95542e-05S10 Sil GLN MET 4.95542e-05S10 Sil GLN ASN 4.95542e-05S10 Sil GLN PRO 4.95542e-05S10 Sil GLN GLN -0.00366811S10 Sil GLN ARG 4.95542e-05S10 Sil GLN SER 4.95542e-05S10 Sil GLN THR 4.95542e-05S10 Sil GLN VAL 4.95542e-05S10 Sil GLN TRP 4.95542e-05S10 Sil GLN TYR 4.95542e-05S10 Sil GLN 4.95542e-05S10 Sil ARG GLN 0.000669087S10 Sil SER GLN 0.000669087S10 Sil THR ALA 9.65074e-06S10 Sil THR CYS 9.65074e-06S10 Sil THR ASP 9.65074e-06S10 Sil THR GLU 9.65074e-06S10 Sil THR PHE 9.65074e-06S10 Sil THR GLY 9.65074e-06S10 Sil THR HIS 9.65074e-06S10 Sil THR ILE 9.65074e-06S10 Sil THR LYS 9.65074e-06S10 Sil THR LEU 9.65074e-06S10 Sil THR MET 9.65074e-06S10 Sil THR ASN 9.65074e-06S10 Sil THR PRO 9.65074e-06S10 Sil THR GLN -0.00175737S10 Sil THR ARG 9.65074e-06S10 Sil THR SER 9.65074e-06S10 Sil THR THR 9.65074e-06S10 Sil THR VAL 9.65074e-06S10 Sil THR TRP 9.65074e-06S10 Sil THR TYR 9.65074e-06S10 Sil THR 9.65074e-06S10 Sil VAL GLN 0.000669087S10 Sil TRP GLN 0.000669087S10 Sil TYR GLN 0.000669087S10 Sil GLN 0.000669087S10 S12 ALA ALA 0.000154741S10 S12 ALA ASP 5.2419e-05S10 S12 ALA HIS 7.34132e-05S10 S12 ALA ASN 1.13851e-06S10 S12 ALA SER 0.000319857S10 S12 ALA THR 6.75156e-05S10 S12 CYS ALA 0.000154741S10 S12 CYS ASP 5.2419e-05S10 S12 CYS HIS 7.34132e-05S10 S12 CYS ASN 1.13851e-06S10 S12 CYS SER 0.000319857S10 S12 CYS THR 6.75156e-05S10 S12 ASP ALA 0.000154741S10 S12 ASP ASP 5.2419e-05S10 S12 ASP HIS 7.34132e-05S10 S12 ASP ASN 1.13851e-06S10 S12 ASP SER 0.000319857S10 S12 ASP THR 6.75156e-05S10 S12 GLU ALA 0.000154741S10 S12 GLU ASP 5.2419e-05S10 S12 GLU HIS 7.34132e-05S10 S12 GLU ASN 1.13851e-06S10 S12 GLU SER 0.000319857S10 S12 GLU THR 6.75156e-05S10 S12 PHE ALA 0.000154741S10 S12 PHE ASP 5.2419e-05S10 S12 PHE HIS 7.34132e-05S10 S12 PHE ASN 1.13851e-06S10 S12 PHE SER 0.000319857S10 S12 PHE THR 6.75156e-05S10 S12 GLY ALA 0.000154741S10 S12 GLY ASP 5.2419e-05S10 S12 GLY HIS 7.34132e-05S10 S12 GLY ASN 1.13851e-06S10 S12 GLY SER 0.000319857S10 S12 GLY THR 6.75156e-05S10 S12 HIS ALA 0.000154741S10 S12 HIS ASP 5.2419e-05S10 S12 HIS HIS 7.34132e-05S10 S12 HIS ASN 1.13851e-06S10 S12 HIS SER 0.000319857S10 S12 HIS THR 6.75156e-05S10 S12 ILE ALA 0.000154741S10 S12 ILE ASP 5.2419e-05S10 S12 ILE HIS 7.34132e-05S10 S12 ILE ASN 1.13851e-06S10 S12 ILE SER 0.000319857S10 S12 ILE THR 6.75156e-05S10 S12 LYS ALA 0.0175394S10 S12 LYS CYS 6.89051e-05S10 S12 LYS ASP 0.00822381S10 S12 LYS GLU 6.89051e-05S10 S12 LYS PHE 6.89051e-05S10 S12 LYS GLY 6.89051e-05S10 S12 LYS HIS -0.0359375S10 S12 LYS ILE 6.89051e-05S10 S12 LYS LYS 6.89051e-05S10 S12 LYS LEU 6.89051e-05S10 S12 LYS MET 6.89051e-05S10 S12 LYS ASN 0.000247286S10 S12 LYS PRO 6.89051e-05S10 S12 LYS GLN 6.89051e-05S10 S12 LYS ARG 6.89051e-05S10 S12 LYS SER -0.0056042S10 S12 LYS THR 0.0124525S10 S12 LYS VAL 6.89051e-05S10 S12 LYS TRP 6.89051e-05S10 S12 LYS TYR 6.89051e-05S10 S12 LYS - 6.89051e-05S10 S12 LEU ALA -0.020466S10 S12 LEU CYS 1.65823e-05S10 S12 LEU ASP 0.00213301S10 S12 LEU GLU 1.65823e-05S10 S12 LEU PHE 1.65823e-05S10 S12 LEU GLY 1.65823e-05S10 S12 LEU HIS 0.00285827S10 S12 LEU ILE 1.65823e-05S10 S12 LEU LYS 1.65823e-05S10 S12 LEU LEU 1.65823e-05S10 S12 LEU MET 1.65823e-05S10 S12 LEU ASN 7.31111e-05S10 S12 LEU PRO 1.65823e-05S10 S12 LEU GLN 1.65823e-05S10 S12 LEU ARG 1.65823e-05S10 S12 LEU SER 0.0110786S10 S12 LEU THR 0.00277143S10 S12 LEU VAL 1.65823e-05S10 S12 LEU TRP 1.65823e-05S10 S12 LEU TYR 1.65823e-05S10 S12 LEU - 1.65823e-05S10 S12 MET ALA -0.0205475S10 S12 MET CYS 0.000449937S10 S12 MET ASP 0.00700104S10 S12 MET GLU 0.000449937S10 S12 MET PHE 0.000449937S10 S12 MET GLY 0.000449937S10 S12 MET HIS 0.0444137S10 S12 MET ILE 0.000449937S10 S12 MET LYS 0.000449937S10 S12 MET LEU 0.000449937S10 S12 MET MET 0.000449937S10 S12 MET ASN -0.000973486S10 S12 MET PRO 0.000449937S10 S12 MET GLN 0.000449937S10 S12 MET ARG 0.000449937S10 S12 MET SER -0.0374405S10 S12 MET THR -0.00231804S10 S12 MET VAL 0.000449937S10 S12 MET TRP 0.000449937S10 S12 MET TYR 0.000449937S10 S12 MET - 0.000449937S10 S12 ASN ALA 0.000154741S10 S12 ASN ASP 5.2419e-05S10 S12 ASN HIS 7.34132e-05S10 S12 ASN ASN 1.13851e-06S10 S12 ASN SER 0.000319857S10 S12 ASN THR 6.75156e-05S10 S12 PRO ALA 0.000154741S10 S12 PRO ASP 5.2419e-05S10 S12 PRO HIS 7.34132e-05S10 S12 PRO ASN 1.13851e-06S10 S12 PRO SER 0.000319857S10 S12 PRO THR 6.75156e-05S10 S12 GLN ALA 0.00838634S10 S12 GLN CYS 0.000181761S10 S12 GLN ASP -0.024409S10 S12 GLN GLU 0.000181761S10 S12 GLN PHE 0.000181761S10 S12 GLN GLY 0.000181761S10 S12 GLN HIS -0.0185644S10 S12 GLN ILE 0.000181761S10 S12 GLN LYS 0.000181761S10 S12 GLN LEU 0.000181761S10 S12 GLN MET 0.000181761S10 S12 GLN ASN 0.000654004S10 S12 GLN PRO 0.000181761S10 S12 GLN GLN 0.000181761S10 S12 GLN ARG 0.000181761S10 S12 GLN SER 0.0504292S10 S12 GLN THR -0.0218995S10 S12 GLN VAL 0.000181761S10 S12 GLN TRP 0.000181761S10 S12 GLN TYR 0.000181761S10 S12 GLN - 0.000181761S10 S12 ARG ALA 0.000154741S10 S12 ARG ASP 5.2419e-05S10 S12 ARG HIS 7.34132e-05S10 S12 ARG ASN 1.13851e-06S10 S12 ARG SER 0.000319857S10 S12 ARG THR 6.75156e-05S10 S12 SER ALA 0.000154741S10 S12 SER ASP 5.2419e-05S10 S12 SER HIS 7.34132e-05S10 S12 SER ASN 1.13851e-06S10 S12 SER SER 0.000319857S10 S12 SER THR 6.75156e-05S10 S12 THR ALA 0.010222S10 S12 THR CYS 2.81142e-05S10 S12 THR ASP 0.00429929S10 S12 THR GLU 2.81142e-05S10 S12 THR PHE 2.81142e-05S10 S12 THR GLY 2.81142e-05S10 S12 THR HIS 0.004088S10 S12 THR ILE 2.81142e-05S10 S12 THR LYS 2.81142e-05S10 S12 THR LEU 2.81142e-05S10 S12 THR MET 2.81142e-05S10 S12 THR ASN 0.000126401S10 S12 THR PRO 2.81142e-05S10 S12 THR GLN 2.81142e-05S10 S12 THR ARG 2.81142e-05S10 S12 THR SER -0.0264844S10 S12 THR THR 0.00576277S10 S12 THR VAL 2.81142e-05S10 S12 THR TRP 2.81142e-05S10 S12 THR TYR 2.81142e-05S10 S12 THR - 2.81142e-05S10 S12 VAL ALA 0.000154741S10 S12 VAL ASP 5.2419e-05S10 S12 VAL HIS 7.34132e-05S10 S12 VAL ASN 1.13851e-06S10 S12 VAL SER 0.000319857S10 S12 VAL THR 6.75156e-05S10 S12 TRP ALA 0.000154741S10 S12 TRP ASP 5.2419e-05S10 S12 TRP HIS 7.34132e-05S10 S12 TRP ASN 1.13851e-06S10 S12 TRP SER 0.000319857S10 S12 TRP THR 6.75156e-05S10 S12 TYR ALA 0.000154741S10 S12 TYR ASP 5.2419e-05S10 S12 TYR HIS 7.34132e-05S10 S12 TYR ASN 1.13851e-06S10 S12 TYR SER 0.000319857S10 S12 TYR THR 6.75156e-05S10 S12 - ALA 0.000154741S10 S12 - ASP 5.2419e-05S10 S12 - HIS 7.34132e-05S10 S12 - ASN 1.13851e-06S10 S12 - SER 0.000319857S10 S12 - THR 6.75156e-05S10 S13 ALA ASP 1.58622e-05S10 S13 ALA LEU 0.000596889S10 S13 ALA PRO 3.26708e-05S10 S13 ALA THR 1.98803e-05S10 S13 ALA TYR 3.78312e-06S10 S13 CYS ASP 1.58622e-05S10 S13 CYS LEU 0.000596889S10 S13 CYS PRO 3.26708e-05S10 S13 CYS THR 1.98803e-05S10 S13 CYS TYR 3.78312e-06S10 S13 ASP ASP 1.58622e-05S10 S13 ASP LEU 0.000596889S10 S13 ASP PRO 3.26708e-05S10 S13 ASP THR 1.98803e-05S10 S13 ASP TYR 3.78312e-06S10 S13 GLU ASP 1.58622e-05S10 S13 GLU LEU 0.000596889S10 S13 GLU PRO 3.26708e-05S10 S13 GLU THR 1.98803e-05S10 S13 GLU TYR 3.78312e-06S10 S13 PHE ASP 1.58622e-05S10 S13 PHE LEU 0.000596889S10 S13 PHE PRO 3.26708e-05S10 S13 PHE THR 1.98803e-05S10 S13 PHE TYR 3.78312e-06S10 S13 GLY ASP 1.58622e-05S10 S13 GLY LEU 0.000596889S10 S13 GLY PRO 3.26708e-05S10 S13 GLY THR 1.98803e-05S10 S13 GLY TYR 3.78312e-06S10 S13 HIS ASP 1.58622e-05S10 S13 HIS LEU 0.000596889S10 S13 HIS PRO 3.26708e-05S10 S13 HIS THR 1.98803e-05S10 S13 HIS TYR 3.78312e-06S10 S13 ILE ASP 1.58622e-05S10 S13 ILE LEU 0.000596889S10 S13 ILE PRO 3.26708e-05S10 S13 ILE THR 1.98803e-05S10 S13 ILE TYR 3.78312e-06S10 S13 LYS ALA 4.0321e-05S10 S13 LYS CYS 4.0321e-05S10 S13 LYS ASP 0.00243108S10 S13 LYS GLU 4.0321e-05S10 S13 LYS PHE 4.0321e-05S10 S13 LYS GLY 4.0321e-05S10 S13 LYS HIS 4.0321e-05S10 S13 LYS ILE 4.0321e-05S10 S13 LYS LYS 4.0321e-05S10 S13 LYS LEU -0.0130401S10 S13 LYS MET 4.0321e-05S10 S13 LYS ASN 4.0321e-05S10 S13 LYS PRO 0.0045701S10 S13 LYS GLN 4.0321e-05S10 S13 LYS ARG 4.0321e-05S10 S13 LYS SER 4.0321e-05S10 S13 LYS THR 0.00280195S10 S13 LYS VAL 4.0321e-05S10 S13 LYS TRP 4.0321e-05S10 S13 LYS TYR 0.000546922S10 S13 LYS 4.0321e-05S10 S13 LEU ALA 9.24212e-06S10 S13 LEU CYS 9.24212e-06S10 S13 LEU ASP 0.000772604S10 S13 LEU GLU 9.24212e-06S10 S13 LEU PHE 9.24212e-06S10 S13 LEU GLY 9.24212e-06S10 S13 LEU HIS 9.24212e-06S10 S13 LEU ILE 9.24212e-06S10 S13 LEU LYS 9.24212e-06S10 S13 LEU LEU -0.00424906S10 S13 LEU MET 9.24212e-06S10 S13 LEU ASN 9.24212e-06S10 S13 LEU PRO 0.00107761S10 S13 LEU GLN 9.24212e-06S10 S13 LEU ARG 9.24212e-06S10 S13 LEU SER 9.24212e-06S10 S13 LEU THR 0.000777521S10 S13 LEU VAL 9.24212e-06S10 S13 LEU TRP 9.24212e-06S10 S13 LEU TYR 0.000170582S10 S13 LEU 9.24212e-06S10 S13 MET ALA 0.000394043S10 S13 MET CYS 0.000394043S10 S13 MET ASP -0.0137826S10 S13 MET GLU 0.000394043S10 S13 MET PHE 0.000394043S10 S13 MET GLY 0.000394043S10 S13 MET HIS 0.000394043S10 S13 MET ILE 0.000394043S10 S13 MET LYS 0.000394043S10 S13 MET LEU -0.00442121S10 S13 MET MET 0.000394043S10 S13 MET ASN 0.000394043S10 S13 MET PRO 0.0282174S10 S13 MET GLN 0.000394043S10 S13 MET ARG 0.000394043S10 S13 MET SER 0.000394043S10 S13 MET THR -0.0159964S10 S13 MET VAL 0.000394043S10 S13 MET TRP 0.000394043S10 S13 MET TYR -0.00343791S10 S13 MET 0.000394043S10 S13 ASN ASP 1.58622e-05S10 S13 ASN LEU 0.000596889S10 S13 ASN PRO 3.26708e-05S10 S13 ASN THR 1.98803e-05S10 S13 ASN TYR 3.78312e-06S10 S13 PRO ASP 1.58622e-05S10 S13 PRO LEU 0.000596889S10 S13 PRO PRO 3.26708e-05S10 S13 PRO THR 1.98803e-05S10 S13 PRO TYR 3.78312e-06S10 S13 GLN ALA 0.000129975S10 S13 GLN CYS 0.000129975S10 S13 GLN ASP 0.00729979S10 S13 GLN GLU 0.000129975S10 S13 GLN PHE 0.000129975S10 S13 GLN GLY 0.000129975S10 S13 GLN HIS 0.000129975S10 S13 GLN ILE 0.000129975S10 S13 GLN LYS 0.000129975S10 S13 GLN LEU 0.0153717S10 S13 GLN MET 0.000129975S10 S13 GLN ASN 0.000129975S10 S13 GLN PRO -0.0379417S10 S13 GLN GLN 0.000129975S10 S13 GLN ARG 0.000129975S10 S13 GLN SER 0.000129975S10 S13 GLN THR 0.0089188S10 S13 GLN VAL 0.000129975S10 S13 GLN TRP 0.000129975S10 S13 GLN TYR 0.00159493S10 S13 GLN 0.000129975S10 S13 ARG ASP 1.58622e-05S10 S13 ARG LEU 0.000596889S10 S13 ARG PRO 3.26708e-05S10 S13 ARG THR 1.98803e-05S10 S13 ARG TYR 3.78312e-06S10 S13 SER ASP 1.58622e-05S10 S13 SER LEU 0.000596889S10 S13 SER PRO 3.26708e-05S10 S13 SER THR 1.98803e-05S10 S13 SER TYR 3.78312e-06S10 S13 THR ALA 1.77088e-05S10 S13 THR CYS 1.77088e-05S10 S13 THR ASP 0.0012439S10 S13 THR GLU 1.77088e-05S10 S13 THR PHE 1.77088e-05S10 S13 THR GLY 1.77088e-05S10 S13 THR HIS 1.77088e-05S10 S13 THR ILE 1.77088e-05S10 S13 THR LYS 1.77088e-05S10 S13 THR LEU -0.0066894S10 S13 THR MET 1.77088e-05S10 S13 THR ASN 1.77088e-05S10 S13 THR PRO 0.00182298S10 S13 THR GLN 1.77088e-05S10 S13 THR ARG 1.77088e-05S10 S13 THR SER 1.77088e-05S10 S13 THR THR 0.00145243S10 S13 THR VAL 1.77088e-05S10 S13 THR TRP 1.77088e-05S10 S13 THR TYR 0.00032251S10 S13 THR 1.77088e-05S10 S13 VAL ASP 1.58622e-05 S10 S13 VAL LEU 0.000596889 S10 S13 VAL PRO 3.26708e-05 S10 S13 VAL THR 1.98803e-05 S10 S13 VAL TYR 3.78312e-06 S10 S13 TRP ASP 1.58622e-05 S10 S13 TRP LEU 0.000596889 S10 S13 TRP PRO 3.26708e-05 S10 S13 TRP THR 1.98803e-05 S10 S13 TRP TYR 3.78312e-06 S10 S13 TYR ASP 1.58622e-05 S10 S13 TYR LEU 0.000596889 S10 S13 TYR PRO 3.26708e-05 S10 S13 TYR THR 1.98803e-05 S10 S13 TYR TYR 3.78312e-06 S10 S13 - ASP 1.58622e-05 S10 S13 - LEU 0.000596889 S10 S13 - PRO 3.26708e-05 S10 S13 - THR 1.98803e-05 S10 S13 - TYR 3.78312e-06 S10 S14 ALA PHE 0.000215118 S10 S14 ALA HIS 2.61752e-05 S10 S14 ALA LYS 6.57248e-05 S10 S14 ALA GLN 0.000259217 S10 S14 ALA ARG 9.35592e-07 S10 S14 ALA SER 2.32654e-05 S10 S14 ALA THR 3.15112e-05 S10 S14 ALA TYR 4.71366e-05 S10 S14 CYS PHE 0.000215118 S10 S14 CYS HIS 2.61752e-05 S10 S14 CYS LYS 6.57248e-05 S10 S14 CYS GLN 0.000259217 S10 S14 CYS ARG 9.35592e-07 S10 S14 CYS SER 2.32654e-05 S10 S14 CYS THR 3.15112e-05 S10 S14 CYS TYR 4.71366e-05 S10 S14 ASP PHE 0.000215118 S10 S14 ASP HIS 2.61752e-05 S10 S14 ASP LYS 6.57248e-05 S10 S14 ASP GLN 0.000259217 S10 S14 ASP ARG 9.35592e-07 S10 S14 ASP SER 2.32654e-05 S10 S14 ASP THR 3.15112e-05 S10 S14 ASP TYR 4.71366e-05 S10 S14 GLU PHE 0.000215118 S10 S14 GLU HIS 2.61752e-05 S10 S14 GLU LYS 6.57248e-05 S10 S14 GLU GLN 0.000259217 S10 S14 GLU ARG 9.35592e-07 S10 S14 GLU SER 2.32654e-05 S10 S14 GLU THR 3.15112e-05 S10 S14 GLU TYR 4.71366e-05 S10 S14 PHE PHE 0.000215118 S10 S14 PHE HIS 2.61752e-05 S10 S14 PHE LYS 6.57248e-05 S10 S14 PHE GLN 0.000259217 S10 S14 PHE ARG 9.35592e-07S10 S14 PHE SER 2.32654e-05S10 S14 PHE THR 3.15112e-05S10 S14 PHE TYR 4.71366e-05S10 S14 GLY PHE 0.000215118S10 S14 GLY HIS 2.61752e-05S10 S14 GLY LYS 6.57248e-05S10 S14 GLY GLN 0.000259217S10 S14 GLY ARG 9.35592e-07S10 S14 GLY SER 2.32654e-05S10 S14 GLY THR 3.15112e-05S10 S14 GLY TYR 4.71366e-05S10 S14 HIS PHE 0.000215118S10 S14 HIS HIS 2.61752e-05S10 S14 HIS LYS 6.57248e-05S10 S14 HIS GLN 0.000259217S10 S14 HIS ARG 9.35592e-07S10 S14 HIS SER 2.32654e-05S10 S14 HIS THR 3.15112e-05S10 S14 HIS TYR 4.71366e-05S10 S14 ILE PHE 0.000215118S10 S14 ILE HIS 2.61752e-05S10 S14 ILE LYS 6.57248e-05S10 S14 ILE GLN 0.000259217S10 S14 ILE ARG 9.35592e-07S10 S14 ILE SER 2.32654e-05S10 S14 ILE THR 3.15112e-05S10 S14 ILE TYR 4.71366e-05S10 S14 LYS ALA 5.84373e-05S10 S14 LYS CYS 5.84373e-05S10 S14 LYS ASP 5.84373e-05S10 S14 LYS GLU 5.84373e-05S10 S14 LYS PHE -0.0153686S10 S14 LYS GLY 5.84373e-05S10 S14 LYS HIS 0.00336875S10 S14 LYS ILE 5.84373e-05S10 S14 LYS LYS 0.0110753S10 S14 LYS LEU 5.84373e-05S10 S14 LYS MET 5.84373e-05S10 S14 LYS ASN 5.84373e-05S10 S14 LYS PRO 5.84373e-05S10 S14 LYS GLN -0.0151343S10 S14 LYS ARG 0.000182653S10 S14 LYS SER 0.00300351S10 S14 LYS THR 0.00431619S10 S14 LYS VAL 5.84373e-05S10 S14 LYS TRP 5.84373e-05S10 S14 LYS TYR 0.00575178S10 S14 LYS - 5.84373e-05S10 S14 LEU ALA 1.68919e-05S10 S14 LEU CYS 1.68919e-05S10 S14 LEU ASP 1.68919e-05S10 S14 LEU GLU 1.68919e-05S10 S14 LEU PHE 0.00724846S10 S14 LEU GLY 1.68919e-05S10 S14 LEU HIS 0.00128423S10 S14 LEU ILE 1.68919e-05S10 S14 LEU LYS 0.0024879S10 S14 LEU LEU 1.68919e-05S10 S14 LEU MET 1.68919e-05S10 S14 LEU ASN 1.68919e-05S10 S14 LEU PRO 1.68919e-05S10 S14 LEU GLN -0.0174534S10 S14 LEU ARG 5.92542e-05S10 S14 LEU SER 0.00125255S10 S14 LEU THR 0.00131393S10 S14 LEU VAL 1.68919e-05S10 S14 LEU TRP 1.68919e-05S10 S14 LEU TYR 0.00228459S10 S14 LEU - 1.68919e-05S10 S14 MET ALA 0.000647877S10 S14 MET CYS 0.000647877S10 S14 MET ASP 0.000647877S10 S14 MET GLU 0.000647877S10 S14 MET PHE 0.041061S10 S14 MET GLY 0.000647877S10 S14 MET HIS -0.0188736S10 S14 MET ILE 0.000647877S10 S14 MET LYS 0.00490876S10 S14 MET LEU 0.000647877S10 S14 MET MET 0.000647877S10 S14 MET ASN 0.000647877S10 S14 MET PRO 0.000647877S10 S14 MET GLN 0.0313316S10 S14 MET ARG -0.0003826S10 S14 MET SER -0.0163618S10 S14 MET THR -0.0223407S10 S14 MET VAL 0.000647877S10 S14 MET TRP 0.000647877S10 S14 MET TYR -0.0308815S10 S14 MET - 0.000647877S10 S14 ASN PHE 0.000215118S10 S14 ASN HIS 2.61752e-05S10 S14 ASN LYS 6.57248e-05S10 S14 ASN GLN 0.000259217S10 S14 ASN ARG 9.35592e-07S10 S14 ASN SER 2.32654e-05S10 S14 ASN THR 3.15112e-05S10 S14 ASN TYR 4.71366e-05S10 S14 PRO PHE 0.000215118S10 S14 PRO HIS 2.61752e-05S10 S14 PRO LYS 6.57248e-05S10 S14 PRO GLN 0.000259217S10 S14 PRO ARG 9.35592e-07S10 S14 PRO SER 2.32654e-05S10 S14 PRO THR 3.15112e-05S10 S14 PRO TYR 4.71366e-05S10 S14 GLN ALA 0.000161327S10 S14 GLN CYS 0.000161327S10 S14 GLN ASP 0.000161327S10 S14 GLN GLU 0.000161327S10 S14 GLN PHE -0.0506112S10 S14 GLN GLY 0.000161327S10 S14 GLN HIS 0.0101134S10 S14 GLN ILE 0.000161327S10 S14 GLN LYS -0.0268228S10 S14 GLN LEU 0.000161327S10 S14 GLN MET 0.000161327S10 S14 GLN ASN 0.000161327S10 S14 GLN PRO 0.000161327S10 S14 GLN GLN 0.0249595S10 S14 GLN ARG 0.000468934S10 S14 GLN SER 0.00839072S10 S14 GLN THR 0.0121293S10 S14 GLN VAL 0.000161327S10 S14 GLN TRP 0.000161327S10 S14 GLN TYR 0.0165975S10 S14 GLN - 0.000161327S10 S14 ARG PHE 0.000215118S10 S14 ARG HIS 2.61752e-05S10 S14 ARG LYS 6.57248e-05S10 S14 ARG GLN 0.000259217S10 S14 ARG ARG 9.35592e-07S10 S14 ARG SER 2.32654e-05S10 S14 ARG THR 3.15112e-05S10 S14 ARG TYR 4.71366e-05S10 S14 SER PHE 0.000215118S10 S14 SER HIS 2.61752e-05S10 S14 SER LYS 6.57248e-05S10 S14 SER GLN 0.000259217S10 S14 SER ARG 9.35592e-07S10 S14 SER SER 2.32654e-05S10 S14 SER THR 3.15112e-05S10 S14 SER TYR 4.71366e-05S10 S14 THR ALA 3.26655e-05S10 S14 THR CYS 3.26655e-05S10 S14 THR ASP 3.26655e-05S10 S14 THR GLU 3.26655e-05S10 S14 THR PHE 0.0117111S10 S14 THR GLY 3.26655e-05S10 S14 THR HIS 0.00234046S10 S14 THR ILE 3.26655e-05S10 S14 THR LYS 0.00564957S10 S14 THR LEU 3.26655e-05S10 S14 THR MET 3.26655e-05S10 S14 THR ASN 3.26655e-05S10 S14 THR PRO 3.26655e-05S10 S14 THR GLN -0.0302602S10 S14 THR ARG 0.00011402S10 S14 THR SER 0.00213868S10 S14 THR THR 0.0025737S10 S14 THR VAL 3.26655e-05S10 S14 THR TRP 3.26655e-05S10 S14 THR TYR 0.00374371S10 S14 THR - 3.26655e-05S10 S14 VAL PHE 0.000215118S10 S14 VAL HIS 2.61752e-05S10 S14 VAL LYS 6.57248e-05S10 S14 VAL GLN 0.000259217S10 S14 VAL ARG 9.35592e-07S10 S14 VAL SER 2.32654e-05S10 S14 VAL THR 3.15112e-05S10 S14 VAL TYR 4.71366e-05S10 S14 TRP PHE 0.000215118S10 S14 TRP HIS 2.61752e-05S10 S14 TRP LYS 6.57248e-05S10 S14 TRP GLN 0.000259217S10 S14 TRP ARG 9.35592e-07S10 S14 TRP SER 2.32654e-05S10 S14 TRP THR 3.15112e-05S10 S14 TRP TYR 4.71366e-05S10 S14 TYR PHE 0.000215118S10 S14 TYR HIS 2.61752e-05S10 S14 TYR LYS 6.57248e-05S10 S14 TYR GLN 0.000259217S10 S14 TYR ARG 9.35592e-07S10 S14 TYR SER 2.32654e-05S10 S14 TYR THR 3.15112e-05S10 S14 TYR TYR 4.71366e-05S10 S14 PHE 0.000215118S10 S14 HIS 2.61752e-05S10 S14 LYS 6.57248e-05S10 S14 GLN 0.000259217S10 S14 ARG 9.35592e-07S10 S14 SER 2.32654e-05S10 S14 THR 3.15112e-05S10 S14 TYR 4.71366e-05S10 S15 ALA PHE 2.23736e-05S10 S15 ALA HIS 0.000208778S10 S15 ALA THR 0.000437933S10 S15 CYS PHE 2.23736e-05S10 S15 CYS HIS 0.000208778S10 S15 CYS THR 0.000437933S10 S15 ASP PHE 2.23736e-05S10 S15 ASP HIS 0.000208778S10 S15 ASP THR 0.000437933S10 S15 GLU PHE 2.23736e-05S10 S15 GLU HIS 0.000208778S10 S15 GLU THR 0.000437933S10 S15 PHE PHE 2.23736e-05S10 S15 PHE HIS 0.000208778S10 S15 PHE THR 0.000437933S10 S15 GLY PHE 2.23736e-05S10 S15 GLY HIS 0.000208778S10 S15 GLY THR 0.000437933S10 S15 HIS PHE 2.23736e-05S10 S15 HIS HIS 0.000208778S10 S15 HIS THR 0.000437933S10 S15 ILE PHE 2.23736e-05S10 S15 ILE HIS 0.000208778S10 S15 ILE THR 0.000437933S10 S15 LYS ALA 3.97772e-05S10 S15 LYS CYS 3.97772e-05S10 S15 LYS ASP 3.97772e-05S10 S15 LYS GLU 3.97772e-05S10 S15 LYS PHE 0.00340837S10 S15 LYS GLY 3.97772e-05S10 S15 LYS HIS -0.0154856S10 S15 LYS ILE 3.97772e-05S10 S15 LYS LYS 3.97772e-05S10 S15 LYS LEU 3.97772e-05S10 S15 LYS MET 3.97772e-05S10 S15 LYS ASN 3.97772e-05S10 S15 LYS PRO 3.97772e-05S10 S15 LYS GLN 3.97772e-05S10 S15 LYS ARG 3.97772e-05S10 S15 LYS SER 3.97772e-05S10 S15 LYS THR 0.00931639S10 S15 LYS VAL 3.97772e-05S10 S15 LYS TRP 3.97772e-05S10 S15 LYS TYR 3.97772e-05S10 S15 LYS 3.97772e-05S10 S15 LEU ALA 7.94894e-06S10 S15 LEU CYS 7.94894e-06S10 S15 LEU ASP 7.94894e-06S10 S15 LEU GLU 7.94894e-06S10 S15 LEU PHE 0.000896898S10 S15 LEU GLY 7.94894e-06S10 S15 LEU HIS 0.00637702S10 S15 LEU ILE 7.94894e-06S10 S15 LEU LYS 7.94894e-06S10 S15 LEU LEU 7.94894e-06S10 S15 LEU MET 7.94894e-06S10 S15 LEU ASN 7.94894e-06S10 S15 LEU PRO 7.94894e-06S10 S15 LEU GLN 7.94894e-06S10 S15 LEU ARG 7.94894e-06S10 S15 LEU SER 7.94894e-06S10 S15 LEU THR -0.00871986S10 S15 LEU VAL 7.94894e-06S10 S15 LEU TRP 7.94894e-06S10 S15 LEU TYR 7.94894e-06S10 S15 LEU 7.94894e-06S10 S15 MET ALA 0.000322203S10 S15 MET CYS 0.000322203S10 S15 MET ASP 0.000322203S10 S15 MET GLU 0.000322203S10 S15 MET PHE -0.0176549S10 S15 MET GLY 0.000322203S10 S15 MET HIS 0.0390947S10 S15 MET ILE 0.000322203S10 S15 MET LYS 0.000322203S10 S15 MET LEU 0.000322203S10 S15 MET MET 0.000322203S10 S15 MET ASN 0.000322203S10 S15 MET PRO 0.000322203S10 S15 MET GLN 0.000322203S10 S15 MET ARG 0.000322203S10 S15 MET SER 0.000322203S10 S15 MET THR -0.0303553S10 S15 MET VAL 0.000322203S10 S15 MET TRP 0.000322203S10 S15 MET TYR 0.000322203S10 S15 MET 0.000322203S10 S15 ASN PHE 2.23736e-05S10 S15 ASN HIS 0.000208778S10 S15 ASN THR 0.000437933S10 S15 PRO PHE 2.23736e-05S10 S15 PRO HIS 0.000208778S10 S15 PRO THR 0.000437933S10 S15 GLN ALA 0.000104651S10 S15 GLN CYS 0.000104651S10 S15 GLN ASP 0.000104651S10 S15 GLN GLU 0.000104651S10 S15 GLN PHE 0.00959263S10 S15 GLN GLY 0.000104651S10 S15 GLN HIS -0.0472124S10 S15 GLN ILE 0.000104651S10 S15 GLN LYS 0.000104651S10 S15 GLN LEU 0.000104651S10 S15 GLN MET 0.000104651S10 S15 GLN ASN 0.000104651S10 S15 GLN PRO 0.000104651S10 S15 GLN GLN 0.000104651S10 S15 GLN ARG 0.000104651S10 S15 GLN SER 0.000104651S10 S15 GLN THR 0.0330588S10 S15 GLN VAL 0.000104651S10 S15 GLN TRP 0.000104651S10 S15 GLN TYR 0.000104651S10 S15 GLN 0.000104651S10 S15 ARG PHE 2.23736e-05S10 S15 ARG HIS 0.000208778S10 S15 ARG THR 0.000437933S10 S15 SER PHE 2.23736e-05S10 S15 SER HIS 0.000208778S10 S15 SER THR 0.000437933S10 S15 THR ALA 1.48777e-05S10 S15 THR CYS 1.48777e-05S10 S15 THR ASP 1.48777e-05S10 S15 THR GLU 1.48777e-05S10 S15 THR PHE 0.00146213S10 S15 THR GLY 1.48777e-05S10 S15 THR HIS 0.010674S10 S15 THR ILE 1.48777e-05S10 S15 THR LYS 1.48777e-05S10 S15 THR LEU 1.48777e-05S10 S15 THR MET 1.48777e-05S10 S15 THR ASN 1.48777e-05S10 S15 THR PRO 1.48777e-05S10 S15 THR GLN 1.48777e-05S10 S15 THR ARG 1.48777e-05S10 S15 THR SER 1.48777e-05S10 S15 THR THR -0.0139682S10 S15 THR VAL 1.48777e-05S10 S15 THR TRP 1.48777e-05S10 S15 THR TYR 1.48777e-05S10 S15 THR 1.48777e-05S10 S15 VAL PHE 2.23736e-05S10 S15 VAL HIS 0.000208778S10 S15 VAL THR 0.000437933S10 S15 TRP PHE 2.23736e-05S10 S15 TRP HIS 0.000208778S10 S15 TRP THR 0.000437933 S10 S15 TYR PHE 2.23736e-05 S10 S15 TYR HIS 0.000208778 S10 S15 TYR THR 0.000437933 S10 S15 - PHE 2.23736e-05 S10 S15 - HIS 0.000208778 S10 S15 - THR 0.000437933 S10 S16 ALA PHE 1.52842e-06 S10 S16 ALA LEU 4.30443e-06 S10 S16 ALA TRP 0.000106084 S10 S16 ALA TYR 0.000557169 S10 S16 CYS PHE 1.52842e-06 S10 S16 CYS LEU 4.30443e-06 S10 S16 CYS TRP 0.000106084 S10 S16 CYS TYR 0.000557169 S10 S16 ASP PHE 1.52842e-06 S10 S16 ASP LEU 4.30443e-06 S10 S16 ASP TRP 0.000106084 S10 S16 ASP TYR 0.000557169 S10 S16 GLU PHE 1.52842e-06 S10 S16 GLU LEU 4.30443e-06 S10 S16 GLU TRP 0.000106084 S10 S16 GLU TYR 0.000557169S10 S16 PHE PHE 1.52842e-06 S10 S16 PHE LEU 4.30443e-06 S10 S16 PHE TRP 0.000106084 S10 S16 PHE TYR 0.000557169 S10 S16 GLY PHE 1.52842e-06 S10 S16 GLY LEU 4.30443e-06 S10 S16 GLY TRP 0.000106084 S10 S16 GLY TYR 0.000557169 S10 S16 HIS PHE 1.52842e-06 S10 S16 HIS LEU 4.30443e-06 S10 S16 HIS TRP 0.000106084 S10 S16 HIS TYR 0.000557169 S10 S16 ILE PHE 1.52842e-06 S10 S16 ILE LEU 4.30443e-06 S10 S16 ILE TRP 0.000106084 S10 S16 ILE TYR 0.000557169 S10 S16 LYS ALA 5.12553e-05 S10 S16 LYS CYS 5.12553e-05 S10 S16 LYS ASP 5.12553e-05 S10 S16 LYS GLU 5.12553e-05 S10 S16 LYS PHE 0.000253162 S10 S16 LYS GLY 5.12553e-05 S10 S16 LYS HIS 5.12553e-05 S10 S16 LYS ILE 5.12553e-05 S10 S16 LYS LYS 5.12553e-05 S10 S16 LYS LEU 0.000771979 S10 S16 LYS MET 5.12553e-05 S10 S16 LYS ASN 5.12553e-05 S10 S16 LYS PRO 5.12553e-05 S10 S16 LYS GLN 5.12553e-05 S10 S16 LYS ARG 5.12553e-05 S10 S16 LYS SER 5.12553e-05 S10 S16 LYS THR 5.12553e-05 S10 S16 LYS VAL 5.12553e-05S10 S16 LYS TRP -0.0270199S10 S16 LYS TYR 0.0230783S10 S16 LYS - 5.12553e-05S10 S16 LEU ALA 1.00737e-05S10 S16 LEU CYS 1.00737e-05S10 S16 LEU ASP 1.00737e-05S10 S16 LEU GLU 1.00737e-05S10 S16 LEU PHE 7.23459e-05S10 S16 LEU GLY 1.00737e-05S10 S16 LEU HIS 1.00737e-05S10 S16 LEU ILE 1.00737e-05S10 S16 LEU LYS 1.00737e-05S10 S16 LEU LEU 0.000225176S10 S16 LEU MET 1.00737e-05S10 S16 LEU ASN 1.00737e-05S10 S16 LEU PRO 1.00737e-05S10 S16 LEU GLN 1.00737e-05S10 S16 LEU ARG 1.00737e-05S10 S16 LEU SER 1.00737e-05S10 S16 LEU THR 1.00737e-05S10 S16 LEU VAL 1.00737e-05S10 S16 LEU TRP 0.00373206S10 S16 LEU TYR -0.00550371S10 S16 LEU - 1.00737e-05S10 S16 MET ALA 0.000254636S10 S16 MET CYS 0.000254636S10 S16 MET ASP 0.000254636S10 S16 MET GLU 0.000254636S10 S16 MET PHE -0.00141513S10 S16 MET GLY 0.000254636S10 S16 MET HIS 0.000254636S10 S16 MET ILE 0.000254636S10 S16 MET LYS 0.000254636S10 S16 MET LEU -0.00441484S10 S16 MET MET 0.000254636S10 S16 MET ASN 0.000254636S10 S16 MET PRO 0.000254636S10 S16 MET GLN 0.000254636S10 S16 MET ARG 0.000254636S10 S16 MET SER 0.000254636S10 S16 MET THR 0.000254636S10 S16 MET VAL 0.000254636S10 S16 MET TRP 0.0609215S10 S16 MET TYR -0.062536S10 S16 MET - 0.000254636S10 S16 ASN PHE 1.52842e-06S10 S16 ASN LEU 4.30443e-06S10 S16 ASN TRP 0.000106084S10 S16 ASN TYR 0.000557169S10 S16 PRO PHE 1.52842e-06S10 S16 PRO LEU 4.30443e-06S10 S16 PRO TRP 0.000106084S10 S16 PRO TYR 0.000557169S10 S16 GLN ALA 0.000120307S10 S16 GLN CYS 0.000120307S10 S16 GLN ASP 0.000120307S10 S16 GLN GLU 0.000120307S10 S16 GLN PHE 0.000593189S10 S16 GLN GLY 0.000120307S10 S16 GLN HIS 0.000120307S10 S16 GLN ILE 0.000120307S10 S16 GLN LYS 0.000120307S10 S16 GLN LEU 0.00185299S10 S16 GLN MET 0.000120307S10 S16 GLN ASN 0.000120307S10 S16 GLN PRO 0.000120307S10 S16 GLN GLN 0.000120307S10 S16 GLN ARG 0.000120307S10 S16 GLN SER 0.000120307S10 S16 GLN THR 0.000120307S10 S16 GLN VAL 0.000120307S10 S16 GLN TRP -0.0470587S10 S16 GLN TYR 0.0398902S10 S16 GLN - 0.000120307S10 S16 ARG PHE 1.52842e-06S10 S16 ARG LEU 4.30443e-06S10 S16 ARG TRP 0.000106084S10 S16 ARG TYR 0.000557169S10 S16 SER PHE 1.52842e-06S10 S16 SER LEU 4.30443e-06S10 S16 SER TRP 0.000106084S10 S16 SER TYR 0.000557169S10 S16 THR ALA 1.53405e-05S10 S16 THR CYS 1.53405e-05S10 S16 THR ASP 1.53405e-05S10 S16 THR GLU 1.53405e-05S10 S16 THR PHE 0.000121226S10 S16 THR GLY 1.53405e-05S10 S16 THR HIS 1.53405e-05S10 S16 THR ILE 1.53405e-05S10 S16 THR LYS 1.53405e-05S10 S16 THR LEU 0.000348047S10 S16 THR MET 1.53405e-05S10 S16 THR ASN 1.53405e-05S10 S16 THR PRO 1.53405e-05S10 S16 THR GLN 1.53405e-05S10 S16 THR ARG 1.53405e-05S10 S16 THR SER 1.53405e-05S10 S16 THR THR 1.53405e-05S10 S16 THR VAL 1.53405e-05S10 S16 THR TRP 0.0052542S10 S16 THR TYR -0.00754863S10 S16 THR - 1.53405e-05S10 S16 VAL PHE 1.52842e-06S10 S16 VAL LEU 4.30443e-06S10 S16 VAL TRP 0.000106084S10 S16 VAL TYR 0.000557169S10 S16 TRP PHE 1.52842e-06S10 S16 TRP LEU 4.30443e-06S10 S16 TRP TRP 0.000106084S10 S16 TRP TYR 0.000557169S10 S16 TYR PHE 1.52842e-06S10 S16 TYR LEU 4.30443e-06S10 S16 TYR TRP 0.000106084S10 S16 TYR TYR 0.000557169S10 S16 - PHE 1.52842e-06S10 S16 - LEU 4.30443e-06S10 S16 - TRP 0.000106084S10 S16 - TYR 0.000557169S10 S17 ALA ASP 0.000162782S10 S17 ALA PHE 0.000370912S10 S17 ALA LYS 1.98803e-05S10 S17 ALA TYR 0.000115511S10 S17 CYS ASP 0.000162782S10 S17 CYS PHE 0.000370912S10 S17 CYS LYS 1.98803e-05S10 S17 CYS TYR 0.000115511S10 S17 ASP ASP 0.000162782S10 S17 ASP PHE 0.000370912S10 S17 ASP LYS 1.98803e-05S10 S17 ASP TYR 0.000115511S10 S17 GLU ASP 0.000162782S10 S17 GLU PHE 0.000370912S10 S17 GLU LYS 1.98803e-05S10 S17 GLU TYR 0.000115511S10 S17 PHE ASP 0.000162782S10 S17 PHE PHE 0.000370912S10 S17 PHE LYS 1.98803e-05S10 S17 PHE TYR 0.000115511S10 S17 GLY ASP 0.000162782S10 S17 GLY PHE 0.000370912S10 S17 GLY LYS 1.98803e-05S10 S17 GLY TYR 0.000115511S10 S17 HIS ASP 0.000162782S10 S17 HIS PHE 0.000370912S10 S17 HIS LYS 1.98803e-05S10 S17 HIS TYR 0.000115511S10 S17 ILE ASP 0.000162782S10 S17 ILE PHE 0.000370912S10 S17 ILE LYS 1.98803e-05S10 S17 ILE TYR 0.000115511S10 S17 LYS ALA 7.03447e-05S10 S17 LYS CYS 7.03447e-05S10 S17 LYS ASP -0.0152585S10 S17 LYS GLU 7.03447e-05S10 S17 LYS PHE 0.0403025S10 S17 LYS GLY 7.03447e-05S10 S17 LYS HIS 7.03447e-05S10 S17 LYS ILE 7.03447e-05S10 S17 LYS LYS 0.0038635S10 S17 LYS LEU 7.03447e-05S10 S17 LYS MET 7.03447e-05S10 S17 LYS ASN 7.03447e-05S10 S17 LYS PRO 7.03447e-05S10 S17 LYS GLN 7.03447e-05S10 S17 LYS ARG 7.03447e-05S10 S17 LYS SER 7.03447e-05S10 S17 LYS THR 7.03447e-05S10 S17 LYS VAL 7.03447e-05S10 S17 LYS TRP 7.03447e-05S10 S17 LYS TYR -0.0321486S10 S17 LYS 7.03447e-05S10 S17 LEU ALA 1.52861e-05S10 S17 LEU CYS 1.52861e-05S10 S17 LEU ASP 0.00568869S10 S17 LEU GLU 1.52861e-05S10 S17 LEU PHE -0.0145567S10 S17 LEU GLY 1.52861e-05S10 S17 LEU HIS 1.52861e-05S10 S17 LEU ILE 1.52861e-05S10 S17 LEU LYS 0.00101187S10 S17 LEU LEU 1.52861e-05S10 S17 LEU MET 1.52861e-05S10 S17 LEU ASN 1.52861e-05S10 S17 LEU PRO 1.52861e-05S10 S17 LEU GLN 1.52861e-05S10 S17 LEU ARG 1.52861e-05S10 S17 LEU SER 1.52861e-05S10 S17 LEU THR 1.52861e-05S10 S17 LEU VAL 1.52861e-05S10 S17 LEU TRP 1.52861e-05S10 S17 LEU TYR 0.00629351S10 S17 LEU 1.52861e-05S10 S17 MET ALA 0.000334373S10 S17 MET CYS 0.000334373S10 S17 MET ASP 0.0680471S10 S17 MET GLU 0.000334373S10 S17 MET PHE -0.0788293S10 S17 MET GLY 0.000334373S10 S17 MET HIS 0.000334373S10 S17 MET ILE 0.000334373S10 S17 MET LYS -0.0186689S10 S17 MET LEU 0.000334373S10 S17 MET MET 0.000334373S10 S17 MET ASN 0.000334373S10 S17 MET PRO 0.000334373S10 S17 MET GLN 0.000334373S10 S17 MET ARG 0.000334373S10 S17 MET SER 0.000334373S10 S17 MET THR 0.000334373S10 S17 MET VAL 0.000334373S10 S17 MET TRP 0.000334373S10 S17 MET TYR 0.0206516S10 S17 MET 0.000334373S10 S17 ASN ASP 0.000162782S10 S17 ASN PHE 0.000370912S10 S17 ASN LYS 1.98803e-05S10 S17 ASN TYR 0.000115511S10 S17 PRO ASP 0.000162782S10 S17 PRO PHE 0.000370912S10 S17 PRO LYS 1.98803e-05S10 S17 PRO TYR 0.000115511S10 S17 GLN ALA 0.000140303S10 S17 GLN CYS 0.000140303S10 S17 GLN ASP -0.0231373S10 S17 GLN GLU 0.000140303S10 S17 GLN PHE 0.0177545S10 S17 GLN GLY 0.000140303S10 S17 GLN HIS 0.000140303S10 S17 GLN ILE 0.000140303S10 S17 GLN LYS 0.00954699S10 S17 GLN LEU 0.000140303S10 S17 GLN MET 0.000140303S10 S17 GLN ASN 0.000140303S10 S17 GLN PRO 0.000140303S10 S17 GLN GLN 0.000140303S10 S17 GLN ARG 0.000140303S10 S17 GLN SER 0.000140303S10 S17 GLN THR 0.000140303S10 S17 GLN VAL 0.000140303S10 S17 GLN TRP 0.000140303S10 S17 GLN TYR -0.0092266S10 S17 GLN 0.000140303S10 S17 ARG ASP 0.000162782S10 S17 ARG PHE 0.000370912S10 S17 ARG LYS 1.98803e-05S10 S17 ARG TYR 0.000115511S10 S17 SER ASP 0.000162782S10 S17 SER PHE 0.000370912S10 S17 SER LYS 1.98803e-05S10 S17 SER TYR 0.000115511S10 S17 THR ALA 3.51564e-05S10 S17 THR CYS 3.51564e-05S10 S17 THR ASP -0.0406169S10 S17 THR GLU 3.51564e-05S10 S17 THR PHE 0.0261455S10 S17 THR GLY 3.51564e-05S10 S17 THR HIS 3.51564e-05S10 S17 THR ILE 3.51564e-05S10 S17 THR LYS 0.00210623S10 S17 THR LEU 3.51564e-05S10 S17 THR MET 3.51564e-05S10 S17 THR ASN 3.51564e-05S10 S17 THR PRO 3.51564e-05S10 S17 THR GLN 3.51564e-05S10 S17 THR ARG 3.51564e-05S10 S17 THR SER 3.51564e-05S10 S17 THR THR 3.51564e-05S10 S17 THR VAL 3.51564e-05S10 S17 THR TRP 3.51564e-05S10 S17 THR TYR 0.0102032S10 S17 THR 3.51564e-05S10 S17 VAL ASP 0.000162782S10 S17 VAL PHE 0.000370912S10 S17 VAL LYS 1.98803e-05S10 S17 VAL TYR 0.000115511S10 S17 TRP ASP 0.000162782S10 S17 TRP PHE 0.000370912S10 S17 TRP LYS 1.98803e-05S10 S17 TRP TYR 0.000115511S10 S17 TYR ASP 0.000162782S10 S17 TYR PHE 0.000370912S10 S17 TYR LYS 1.98803e-05S10 S17 TYR TYR 0.000115511S10 S17 ASP 0.000162782S10 S17 - PHE 0.000370912S10 S17 - LYS 1.98803e-05S10 517 - TYR 0.000115511S10 518 ALA ALA 0.000112054S10 SI 8 ALA ILE 6.16972e-05S10 SI 8 ALA ASN 9.85341e-05S10 SI 8 ALA GLN 1.98803e-05S10 SI 8 ALA SER 0.000112672S10 SI 8 ALA THR 0.000264247S10 SI 8 CYS ALA 0.000112054S10 S18 CYS ILE 6.16972e-05S10 SI 8 CYS ASN 9.85341e-05S10 SI 8 CYS GLN 1.98803e-05S10 S18 CYS SER 0.000112672S10 SI 8 CYS THR 0.000264247S10 SI 8 ASP ALA 0.000112054S10 S18 ASP ILE 6.16972e-05S10 S18 ASP ASN 9.85341e-05S10 S18 ASP GLN 1.98803e-05S10 S18 ASP SER 0.000112672S10 SI 8 ASP THR 0.000264247S10 SI 8 GLU ALA 0.000112054S10 S18 GLU ILE 6.16972e-05S10 SI 8 GLU ASN 9.85341e-05S10 SI 8 GLU GLN 1.98803e-05S10 SI 8 GLU SER 0.000112672S10 SI 8 GLU THR 0.000264247S10 SI 8 PHE ALA 0.000112054S10 S18 PHE ILE 6.16972e-05S10 SI 8 PHE ASN 9.85341e-05S10 SI 8 PHE GLN 1.98803e-05S10 SI 8 PHE SER 0.000112672S10 SI 8 PHE THR 0.000264247S10 SI 8 GLY ALA 0.000112054S10 S18 GLY ILE 6.16972e-05S10 SI 8 GLY ASN 9.85341e-05S10 SI 8 GLY GLN 1.98803e-05S10 S18 GLY SER 0.000112672S10 SI 8 GLY THR 0.000264247S10 S18 HIS ALA 0.000112054S10 S18 HIS ILE 6.16972e-05S10 S18 HIS ASN 9.85341e-05S10 S18 HIS GLN 1.98803e-05S10 S18 HIS SER 0.000112672S10 S18 HIS THR 0.000264247S10 SI 8 ILE ALA 0.000112054S10 S18 ILE ILE 6.16972e-05S10 S18 ILE ASN 9.85341e-05S10 S18 ILE GLN 1.98803e-05S10 S18 ILE SER 0.000112672S10 S18 ILE THR 0.000264247S10 SI 8 LYS ALA 0.0163878S10 S18 LYS CYS 7.75889e-05S10 S18 LYS ASP 7.75889e-05S10 SI 8 LYS GLU 7.75889e-05S10 SI 8 LYS PHE 7.75889e-05S10 SI 8 LYS GLY 7.75889e-05S10 S18 LYS HIS 7.75889e-05S10 S18 LYS ILE 0.00921064S10 S18 LYS LYS 7.75889e-05S10 S18 LYS LEU 7.75889e-05S10 S18 LYS MET 7.75889e-05S10 S18 LYS ASN -0.0287633S10 S18 LYS PRO 7.75889e-05S10 S18 LYS GLN 0.00345152S10 S18 LYS ARG 7.75889e-05S10 S18 LYS SER -0.031161S10 S18 LYS THR 0.0276655S10 S18 LYS VAL 7.75889e-05S10 S18 LYS TRP 7.75889e-05S10 S18 LYS TYR 7.75889e-05S10 S18 LYS - 7.75889e-05S10 S18 LEU ALA 0.00481817S10 S18 LEU CYS 1.70807e-05S10 S18 LEU ASP 1.70807e-05S10 S18 LEU GLU 1.70807e-05S10 S18 LEU PHE 1.70807e-05S10 S18 LEU GLY 1.70807e-05S10 S18 LEU HIS 1.70807e-05S10 S18 LEU ILE 0.00336634S10 S18 LEU LYS 1.70807e-05S10 S18 LEU LEU 1.70807e-05S10 S18 LEU MET 1.70807e-05S10 S18 LEU ASN 0.00379397S10 S18 LEU PRO 1.70807e-05S10 S18 LEU GLN 0.000917363S10 S18 LEU ARG 1.70807e-05S10 S18 LEU SER 0.00380239S10 S18 LEU THR -0.0182573S10 S18 LEU VAL 1.70807e-05S10 S18 LEU TRP 1.70807e-05S10 S18 LEU TYR 1.70807e-05S10 S18 LEU - 1.70807e-05S10 S18 MET ALA 0.0244565S10 S18 MET CYS 0.000485956S10 S18 MET ASP 0.000485956S10 S18 MET GLU 0.000485956S10 S18 MET PHE 0.000485956S10 S18 MET GLY 0.000485956S10 S18 MET HIS 0.000485956S10 S18 MET ILE 0.00912724S10 S18 MET LYS 0.000485956S10 S18 MET LEU 0.000485956S10 S18 MET MET 0.000485956S10 S18 MET ASN 0.0172733S10 S18 MET PRO 0.000485956S10 S18 MET GLN -0.0175811S10 S18 MET ARG 0.000485956S10 S18 MET SER 0.0215206S10 S18 MET THR -0.0652018S10 S18 MET VAL 0.000485956S10 S18 MET TRP 0.000485956S10 S18 MET TYR 0.000485956S10 S18 MET - 0.000485956S10 S18 ASN ALA 0.000112054S10 S18 ASN ILE 6.16972e-05S10 S18 ASN ASN 9.85341e-05S10 S18 ASN GLN 1.98803e-05S10 S18 ASN SER 0.000112672S10 S18 ASN THR 0.000264247S10 S18 PRO ALA 0.000112054S10 S18 PRO ILE 6.16972e-05S10 S18 PRO ASN 9.85341e-05S10 S18 PRO GLN 1.98803e-05S10 S18 PRO SER 0.000112672S10 S18 PRO THR 0.000264247S10 S18 GLN ALA -0.00340432S10 S18 GLN CYS 0.000192319S10 S18 GLN ASP 0.000192319S10 S18 GLN GLU 0.000192319S10 S18 GLN PHE 0.000192319S10 S18 GLN GLY 0.000192319S10 S18 GLN HIS 0.000192319S10 S18 GLN ILE -0.0289844S10 S18 GLN LYS 0.000192319S10 S18 GLN LEU 0.000192319S10 S18 GLN MET 0.000192319S10 S18 GLN ASN -0.00293703S10 S18 GLN PRO 0.000192319S10 S18 GLN GLN 0.00961149S10 S18 GLN ARG 0.000192319S10 S18 GLN SER -0.00713292S10 S18 GLN THR 0.0272853S10 S18 GLN VAL 0.000192319S10 S18 GLN TRP 0.000192319S10 S18 GLN TYR 0.000192319S10 S18 GLN 0.000192319S10 S18 ARG ALA 0.000112054S10 S18 ARG ILE 6.16972e-05S10 S18 ARG ASN 9.85341e-05S10 S18 ARG GLN 1.98803e-05S10 S18 ARG SER 0.000112672S10 S18 ARG THR 0.000264247S10 S18 SER ALA 0.000112054S10 S18 SER ILE 6.16972e-05S10 S18 SER ASN 9.85341e-05S10 S18 SER GLN 1.98803e-05S10 S18 SER SER 0.000112672S10 S18 SER THR 0.000264247S10 S18 THR ALA -0.046074S10 S18 THR CYS 3.8468e-05S10 S18 THR ASP 3.8468e-05S10 S18 THR GLU 3.8468e-05S10 S18 THR PHE 3.8468e-05S10 S18 THR GLY 3.8468e-05S10 S18 THR HIS 3.8468e-05S10 S18 THR ILE 0.00451526S10 S18 THR LYS 3.8468e-05S10 S18 THR LEU 3.8468e-05S10 S18 THR MET 3.8468e-05S10 S18 THR ASN 0.00700826S10 S18 THR PRO 3.8468e-05S10 S18 THR GLN 0.00185767S10 S18 THR ARG 3.8468e-05S10 S18 THR SER 0.00900898S10 S18 THR THR 0.0215424S10 S18 THR VAL 3.8468e-05S10 S18 THR TRP 3.8468e-05S10 S18 THR TYR 3.8468e-05S10 S18 THR 3.8468e-05S10 S18 VAL ALA 0.000112054S10 S18 VAL ILE 6.16972e-05S10 S18 VAL ASN 9.85341e-05S10 S18 VAL GLN 1.98803e-05S10 S18 VAL SER 0.000112672S10 S18 VAL THR 0.000264247S10 S18 TRP ALA 0.000112054S10 S18 TRP ILE 6.16972e-05S10 S18 TRP ASN 9.85341e-05S10 S18 TRP GLN 1.98803e-05S10 S18 TRP SER 0.000112672S10 S18 TRP THR 0.000264247S10 S18 TYR ALA 0.000112054S10 S18 TYR ILE 6.16972e-05S10 S18 TYR ASN 9.85341e-05S10 S18 TYR GLN 1.98803e-05S10 S18 TYR SER 0.000112672S10 S18 TYR THR 0.000264247S10 S18 ALA 0.000112054S10 S18 ILE 6.16972e-05S10 S18 ASN 9.85341e-05S10 S18 GLN 1.98803e-05S10 S18 SER 0.000112672S10 S18 THR 0.000264247S10 S19 ALA PHE 0.000638484S10 S19 ALA GLN 1.47385e-05S10 S19 ALA TRP 1.58622e-05S10 S19 CYS PHE 0.000638484S10 S19 CYS GLN 1.47385e-05S10 S19 CYS TRP 1.58622e-05S10 S19 ASP PHE 0.000638484S10 S19 ASP GLN 1.47385e-05S10 S19 ASP TRP 1.58622e-05S10 S19 GLU PHE 0.000638484S10 S19 GLU GLN 1.47385e-05S10 S19 GLU TRP 1.58622e-05S10 S19 PHE PHE 0.000638484S10 S19 PHE GLN 1.47385e-05S10 S19 PHE TRP 1.58622e-05S10 S19 GLY PHE 0.000638484S10 S19 GLY GLN 1.47385e-05S10 S19 GLY TRP 1.58622e-05S10 S19 HIS PHE 0.000638484S10 S19 HIS GLN 1.47385e-05S10 S19 HIS TRP 1.58622e-05S10 S19 ILE PHE 0.000638484S10 S19 ILE GLN 1.47385e-05S10 S19 ILE TRP 1.58622e-05S10 S19 LYS ALA 3.04788e-05S10 S19 LYS CYS 3.04788e-05S10 S19 LYS ASP 3.04788e-05S10 S19 LYS GLU 3.04788e-05S10 S19 LYS PHE -0.00685325S10 S19 LYS GLY 3.04788e-05S10 S19 LYS HIS 3.04788e-05S10 S19 LYS ILE 3.04788e-05S10 S19 LYS LYS 3.04788e-05S10 S19 LYS LEU 3.04788e-05S10 S19 LYS MET 3.04788e-05S10 S19 LYS ASN 3.04788e-05S10 S19 LYS PRO 3.04788e-05S10 S19 LYS GLN 0.0018525S10 S19 LYS ARG 3.04788e-05S10 S19 LYS SER 3.04788e-05S10 S19 LYS THR 3.04788e-05S10 S19 LYS VAL 3.04788e-05S10 S19 LYS TRP 0.00240663S10 S19 LYS TYR 3.04788e-05S10 S19 LYS - 3.04788e-05S10 S19 LEU ALA 1.1056e-05S10 S19 LEU CYS 1.1056e-05S10 S19 LEU ASP 1.1056e-05S10 S19 LEU GLU 1.1056e-05S10 S19 LEU PHE -0.00343151S10 S19 LEU GLY 1.1056e-05S10 S19 LEU HIS 1.1056e-05S10 S19 LEU ILE 1.1056e-05S10 S19 LEU LYS 1.1056e-05S10 S19 LEU LEU 1.1056e-05S10 S19 LEU MET 1.1056e-05S10 S19 LEU ASN 1.1056e-05S10 S19 LEU PRO 1.1056e-05S10 S19 LEU GLN 0.000950382S10 S19 LEU ARG 1.1056e-05S10 S19 LEU SER 1.1056e-05S10 S19 LEU THR 1.1056e-05S10 S19 LEU VAL 1.1056e-05S10 S19 LEU TRP 0.000979249S10 S19 LEU TYR 1.1056e-05S10 S19 LEU - 1.1056e-05S10 S19 MET ALA 0.00029357S10 S19 MET CYS 0.00029357S10 S19 MET ASP 0.00029357S10 S19 MET GLU 0.00029357S10 S19 MET PHE 0.0164157S10 S19 MET GLY 0.00029357S10 S19 MET HIS 0.00029357S10 S19 MET ILE 0.00029357S10 S19 MET LYS 0.00029357S10 S19 MET LEU 0.00029357S10 S19 MET MET 0.00029357S10 S19 MET ASN 0.00029357S10 S19 MET PRO 0.00029357S10 S19 MET GLN -0.0110221S10 S19 MET ARG 0.00029357S10 S19 MET SER 0.00029357S10 S19 MET THR 0.00029357S10 S19 MET VAL 0.00029357S10 S19 MET TRP -0.0137952S10 S19 MET TYR 0.00029357S10 S19 MET - 0.00029357S10 S19 ASN PHE 0.000638484S10 S19 ASN GLN 1.47385e-05S10 S19 ASN TRP 1.58622e-05S10 S19 PRO PHE 0.000638484S10 S19 PRO GLN 1.47385e-05S10 S19 PRO TRP 1.58622e-05S10 S19 GLN ALA 8.17503e-05S10 S19 GLN CYS 8.17503e-05S10 S19 GLN ASP 8.17503e-05S10 S19 GLN GLU 8.17503e-05S10 S19 GLN PHE -0.015948S10 S19 GLN GLY 8.17503e-05S10 S19 GLN HIS 8.17503e-05S10 S19 GLN ILE 8.17503e-05S10 S19 GLN LYS 8.17503e-05S10 S19 GLN LEU 8.17503e-05S10 S19 GLN MET 8.17503e-05S10 S19 GLN ASN 8.17503e-05S10 S19 GLN PRO 8.17503e-05S10 S19 GLN GLN 0.00519373S10 S19 GLN ARG 8.17503e-05S10 S19 GLN SER 8.17503e-05S10 S19 GLN THR 8.17503e-05S10 S19 GLN VAL 8.17503e-05S10 S19 GLN TRP 0.00660556S10 S19 GLN TYR 8.17503e-05S10 S19 GLN - 8.17503e-05S10 S19 ARG PHE 0.000638484S10 S19 ARG GLN 1.47385e-05S10 S19 ARG TRP 1.58622e-05S10 S19 SER PHE 0.000638484S10 S19 SER GLN 1.47385e-05S10 S...

Claims

CLAIMSWhat is claimed is:

1. A polypeptide comprising: a) an immunoglobulin heavy chain variable domain (VH) amino acid sequence comprising a heavy chain complementarity determining region 1 (HCDR1), a heavy chain complementarity determining region 2 (HCDR2), and a heavy chain complementarity determining region 3 (HCDR3) that are substantially similar to an HCDR1, HCDR2 and HCDR3, respectively, of the amino acid sequence of any one of SEQ ID NOs: 18, 6-17, 19, and 20; and b) an immunoglobulin light chain variable domain (VL) amino acid sequence comprising a light chain complementarity determining region 1 (LCDR1), a light chain complementarity determining region 2 (LCDR2), and a light chain complementarity determining region 3 (LCDR3) that are substantially similar to an LCDR1, LCDR2 and LCDR3, respectively, of the amino acid sequence of any one of SEQ ID NOs:37, 25-36, 38, and 39.

2. The polypeptide of claim 1, comprising the HCDR1, HCDR2 and HCDR3, and LCDR1, LCDR2 and LCDR3, of an antibody comprising an amino acid sequence selected from:SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); orSEQ ID NO:20 and SEQ ID NO:39 (AB-15c).

3. A polypeptide comprising a paratope that is substantially similar to a paratope of an antibody comprising a VH and VL pair selected from:SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NOV and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c); or SEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or a combination of any of the foregoing.

4. A polypeptide comprising an immunoglobulin heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:4, wherein:Xi is not T;X2 is not R;X3 is not G;X4 is notN;X5 is not T;Xe is not K;X7 is not R;Xs is not L;Xgis not S;Xiois not I;Xu is not T;X12 is not I;X13 is not R;Xi4 is not R;X15 is not Y; orXi6 is not V; or any combination of the foregoing.

5. The polypeptide of claim 4, comprising an immunoglobulin light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:23, wherein:X17 is not L;Xis is not I;X19 is not Y;X20 is not G;X21 is not A;X22 is not Q; orX23 is not T; or any combination of the foregoing.

6. A polypeptide comprising an immunoglobulin heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:4, wherein:Xi is T or N;X2 is R, S or K;X3is G, S, D or T;X4is N, T or K;X5is T, K or I;Xe is K or R;X7 is R or A;Xs is L, W or Y;X9is S or T;X10 is I, A, K, T, S or V;Xn is T, A, K, S or R;X12 is I, V, A or T;X13 is R or K;X14 is R, Y, E, I, N, S, V or K;X15 is Y, V or R; orXi6 is V or I; or any combination of the foregoing.

7. The polypeptide of claim 6, wherein:Xi is N;X2 is S or K;X3is S, D or T;X4is T or K;X5is K or I;Xe is R;X7 is A;Xs is W or Y;X9is T;X10 is A, K, T, S or V;Xn is A, K, S or R;X12 is V, A or T;X13 is K;X14 is Y, E, I, N, S, V or K;X15 is V or R; orXi6 is I; or any combination of the foregoing.

8. The polypeptide of claim 6 or 7, comprising an immunoglobulin light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:23, wherein:X17 is L or D;Xis is I, Y, S, T, E, L, V, Q, H or R;X19 is Y or K;X20 is G or A;X21 is A, S, T or D;X22 is Q, S, K, T, F, H or Y; orX23 is T or H; or any combination of the foregoing.

9. The polypeptide of claim 8, wherein:X17 is D;Xis is Y, S, T, E, L, V, Q, H or R;X19 is K;X20 is A;X21 is S, T or D;X22 is S, K, T, F, H or Y; orX23 is H; or any combination of the foregoing.

10. The polypeptide of any one of claims 1-9, wherein the VH comprises a heavy chain complementarity determining region 1 (HCDR1), a heavy chain complementarity determining region 2 (HCDR2) and a heavy chain complementarity determining region 3 (HCDR3) that are identical in amino acid sequence to the HCDR1, HCDR2 and HCDR3, respectively, of any one of SEQ ID NOs:6-20.

11. The polypeptide of any one of claims 1-10, wherein the VL comprises a light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2) and light chain complementarity determining region 3 (LCDR3) that are identical in amino acid sequence to the LCDR1, LCDR2 and LCDR3, respectively, of any one of SEQ ID NOs:25-39.

12. The polypeptide of any one of claims 1-11, wherein the VH has at least 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:6-20.

13. The polypeptide of any one of claims 1-12, wherein the VH comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of any one or more of SEQ ID NOs:6-20.

14. The polypeptide of any one of claims 1-13, wherein the VL has at least 85% sequence identity to the amino acid sequence of any one or more of SEQ ID NOs:25-39.

15. The polypeptide of any one of claims 1-14, wherein the VL comprises about 1-10 amino acid substitutions, relative to the amino acid sequence of any one or more of SEQ ID NOs:25-39.

16. The polypeptide of claim 13 or 15, wherein the amino acid substitutions are conservative substitutions.

17. The polypeptide of claim 16, wherein the amino acid substitutions are highly conservative substitutions.

18. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO:6; and b) the VL comprises the amino acid sequence of SEQ ID NO:25 (AB-1).

19. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO:7; and b) the VL comprises the amino acid sequence of SEQ ID NO:26 (AB-2).

20. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO:8; and b) the VL comprises the amino acid sequence of SEQ ID NO:27 (AB-3).

21. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NOV; and b) the VL comprises the amino acid sequence of SEQ ID NO:28 (AB-4).

22. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 10; and b) the VL comprises the amino acid sequence of SEQ ID NO:29 (AB-5).

23. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 11; and b) the VL comprises the amino acid sequence of SEQ ID NO:30 (AB-6).

24. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 12; and b) the VL comprises the amino acid sequence of SEQ ID NO:31 (AB-7).

25. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 13; and b) the VL comprises the amino acid sequence of SEQ ID NO:32 (AB-8).

26. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 14; and b) the VL comprises the amino acid sequence of SEQ ID NO:33 (AB-9).

27. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 15; and b) the VL comprises the amino acid sequence of SEQ ID NO:34 (AB-10).

28. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 16; and b) the VL comprises the amino acid sequence of SEQ ID NO:35 (AB-11).

29. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 17; and b) the VL comprises the amino acid sequence of SEQ ID NO:36 (AB-12).

30. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 18; and b) the VL comprises the amino acid sequence of SEQ ID NO:37 (AB-13).

31. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO: 19; and b) the VL comprises the amino acid sequence of SEQ ID NO:38 (AB-14c).

32. The polypeptide of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence of SEQ ID NO:20; and b) the VL comprises the amino acid sequence of SEQ ID NO:39 (AB-15c).

33. The polypeptide of any one of claims 1-32, wherein the VH and VL are humanized, contain human framework regions, or any combination of the foregoing.

34. The polypeptide of any of the preceding claims, wherein the polypeptide binds an interleukin-4 receptor alpha (IL-4Ra).

35. The polypeptide of any one of claims 1-34, wherein the polypeptide is an antibody or an antigen-binding fragment thereof.

36. The polypeptide of claim 35, wherein the antigen binding fragment is selected from Fab, F(ab’)2, Fab’, scFv, or Fv.

37. The polypeptide of claim 35, comprising an antibody heavy chain constant domain sequence, an antibody light chain constant domain sequence, or both an antibody heavy chain constant domain sequence and an antibody light chain constant domain sequence.

38. The polypeptide of claim 37, wherein the antibody heavy chain constant domain is selected from the group consisting of an IgA constant domain, an IgD constant domain, an IgE constant domain, an IgG constant domain and an IgM constant domain.

39. The polypeptide of claim 38, wherein the antibody heavy chain constant domain is an IgG4 heavy chain constant domain.

40. The polypeptide of any one of claims 37-39, comprising an antibody light chain constant domain selected from the group consisting of a K constant domain or a constant domain.

41. The polypeptide of claim 40, wherein the antibody light chain constant domain is a K light chain constant domain.

42. The polypeptide of any one of claims 1-41, wherein the polypeptide is conjugated to a heterologous moiety.

43. The polypeptide of claim 42, wherein the heterologous moiety is a therapeutic agent, a diagnostic agent, or any combination of the foregoing.

44. The polypeptide of claim 42, wherein the heterologous moiety is polyethylene glycol (PEG), hexadecanoic acid, a hydrogel, a lipid nanoparticle, a polymer nanoparticle, a heterologous polypeptide sequence, or any combination of the foregoing.

45. The polypeptide of claim 44, wherein the polymer nanoparticle comprises poly(lactic- co-glycolic acid) (PLGA).

46. The polypeptide of claim 42, wherein the heterologous polypeptide sequence comprises a carrier polypeptide.

47. The polypeptide of claim 46, wherein the carrier polypeptide is albumin or an Fc polypeptide.

48. The polypeptide of any one of claims 1-47, wherein the IL-4Ra comprises one of SEQ ID NOs: 1-3.

49. The polypeptide of claim 48, wherein the polypeptide: a) binds IL-4Ra with a KD of about 1 pM or less; b) binds IL-4Ra with a kaof about 100 x 105M^s'1or less; c) dissociates from IL-4Ra with a kd of about 100 x 10'5s'1or less; d) binds IL-4Ra with an ECso of about 1 pM or less; e) blocks IL-4 Type II signaling with an ICso of about 1 pM or less; f) blocks IL- 13 Type II signaling with an ICso of about 1 pM or less; g) inhibits CD23 expression in B cells with an ICso of about 1 pM or less; or any combination of the foregoing.

50. The polypeptide of claim 48, wherein the polypeptide: a) binds IL-4Ra with a KD of about 0.05 to 0.5 nM or less; b) binds IL-4Ra with a kaof about 7 x 105to 9 x 105M^s'1or less; c) dissociates from IL-4Ra with a kd of about 2 x 10'5to 3 x 10'5s'1or less; d) binds IL-4Ra with an ECso of about 0.02 to 1.5 nM or less; e) blocks IL-4 Type II signaling with an ICso of about 1.1 to 7.3 nM or less; f) blocks IL- 13 Type II signaling with an ICso of about 3 to 3.3 nM or less; g) inhibits CD23 expression in B cells with an ICso of about 8 to 50nM or less; or any combination of the foregoing.

51. The polypeptide of claim 48, wherein the polypeptide: a) binds IL-4Ra with a KD of about 0.02 to 0.04 nM or less; b) binds IL-4Ra with a kaof about 7.7 x 105to 8.9 x 105M^s'1or less; c) dissociates from IL-4Ra with a kd of about 2.1 x 10'5to 2.6 x 10'5s'1or less; d) binds IL-4Ra with an ECso of about 0.04 to 1.3 nM or less; e) blocks IL-4 Type II signaling with an ICso of about 4.4 to 7.3 nM or less;f) inhibits CD23 expression in B cells with an IC50 of about 30 to 50 nM or less; or any combination of the foregoing.

52. The polypeptide of claim 48, wherein the polypeptide: a) blocks IL-4 Type II signaling with an IC50 of about 1.1 to 1.8 nM or less; b) inhibits CD23 expression in B cells with an IC50 of about 30 to 34 nM or less; or both.

53. The polypeptide of claim 48, wherein the polypeptide inhibits CD23 expression in B cells with an IC50 of about 8 to 8.8 nM or less.

54. A fusion protein comprising the polypeptide of any one of claims 1-53.

55. A polynucleotide (e.g., DNA or RNA; linear or circular; optionally containing one or more modified nucleotides) comprising a sequence encoding the polypeptide of any one of claims 1-53 or the fusion protein of claim 54.

56. A vector (e.g., an expression vector, including a viral-delivery vector) comprising the polynucleotide of claim 55.

57. A host cell comprising the polynucleotide of claim 55 or the vector of claim 56.

58. A composition comprising the polypeptide of any one of claims 1-53 or the fusion protein of claim 54 or the polynucleotide of claim 55.

59. The composition of claim 58, comprising one or more pharmaceutical excipients, diluents, or carriers.

60. A method of treating a subject in need thereof, comprising administering an effective amount of the composition of claim 58 or 59 to the subject.

61. A method of reducing inflammation, a symptom of an inflammatory condition, or risk of developing an inflammatory condition in a cell of a subject, comprising contacting the cell with an effective amount of the composition of claim 58 or 59.

62. The method of claim 60 or 61, wherein the subject has, is suspected of having, or is at risk of developing an inflammatory condition.

63. The method of claim 62, wherein the inflammatory condition is selected from the group comprising asthma, eczema (atopic dermatitis), food allergy, prurigo nodularis, chronic rhinosinusitis with nasal polyps, keloids, eosinophilic esophagitis, prostate cancer, chronic urticaria, bullous pemphigoid, localized scleroderma, alopecia areata, ulcerative colitis, aspirin-exacerbated respiratory disease, metastatic non-small cell lung cancer, Netherton syndrome, and combinations thereof.

64. The method of any one of claims 60-63, wherein the subject has an atopic dermatitis.

65. The method of any one of claims 60-64, wherein the subject is a human.

66. The method of any one of claims 60-65, comprising administering a therapeutically effective amount of an additional therapeutic or prophylactic agent to the subject.

67. The method of claim 66, wherein the additional therapeutic or prophylactic agent is selected from the group comprising an additional anti-IL-4Ra antibody or antigenbinding fragment thereof, an inhaled corticosteroid, a leukotriene modifier, a combination corticosteroid / long-acting beta agonist inhaler, a bronchodilator, a shortacting beta agonist, an anticholinergic agent, an oral or intravenous corticosteroid, a calcineurin inhibitor, an anti-inflammatory agent, an anti-rheumatic agent, an immunosuppressant, a biologic agent, an antihistamine, an epinephrine, an oral immunotherapy, a topical corticosteroid, a topical anesthetic, an immunomodulatory agent, a nasal corticosteroid, an antifungal agent, an anti-androgen agent, a luteinizing hormone-releasing hormone (LHRH) agonist, an gonadotropin-releasing hormone (GnRH) antagonist, a chemotherapy, a targeted drug therapy, a steroid-sparing agent, and combinations thereof.

68. The method of any one of claims 60-67, wherein the subject previously received a therapeutic or prophylactic agent.

69. A computer-implemented method for predicting a functional property of a polypeptide, the method comprising, via a computationally binding optimized (CBO) model: for each amino acid position of an amino acid sequence of the polypeptide:determining a plurality of energy scores based on the amino acid position in the amino acid sequence; generating a partition function based on the plurality of energy scores determined; and determining a cross-entropy metric based on (i) an amino acid at the amino acid position in the amino acid sequence, (ii) a maximum energy score of the plurality of energy scores determined, and (iii) the generated partition function; and generating an analysis score of the polypeptide based on each cross-entropy metric determined, the analysis score indicating a predicted functional property of the polypeptide, wherein the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 37, and wherein the polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.

70. The computer-implemented method of claim 69, wherein a given plurality of energy scores include at least one of: a single amino acid energy score and a pairwise amino acid energy score.

71. The computer-implemented method of claim 69, wherein for at least one amino acid position of the amino acid sequence of the polypeptide, determining the plurality of energy scores is further based on having substituted the amino acid at the amino acid position in the amino acid sequence with each of a plurality of different amino acids.

72. The computer-implemented method of claim 69, wherein for at least one amino acid position of the amino acid sequence of the polypeptide, generating the partition function is further based on a softmax function.

73. The computer-implemented method of claim 69, wherein at least one of:predicting the functional property of the polypeptide is implementable by a script of Appendix A or Appendix B, and the CBO model is substantially similar to a table of Appendix C.

74. The computer-implemented method of claim 69, wherein the generated analysis score is above a threshold, and wherein the threshold is a score from the CBO model of one or more of a reference polypeptide that includes a VH and VL pair selected from:SEQ ID NO:5 and SEQ ID NO:24 (Reference);SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c);SEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or a combination of any of the foregoing.

75. The computer-implemented method of claim 69, wherein the predicted functional property is a binding affinity for interleukin-4 receptor alpha (IL-4Ra).

76. The computer-implemented method of claim 69, wherein the predicted functional property is at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 1 pM or less, a binding affinity for IL-4Ra characterized by a kaof about 100 x 105T's-1or less,a dissociation from IL-4Ra characterized by a kd of about 100 x 10’5s’1or less, a binding affinity for IL-4Ra characterized by an ECso of about 1 pM or less, a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1 pM or less, a blocking activity against IL- 13 Type II signaling characterized by an IC50 of about 1 pM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 1 pM or less.

77. The computer-implemented method of claim 69, wherein the predicted functional property is at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.05 to 0.5 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7 x 105to 9 x 105T's-1or less, a dissociation from IL-4Ra characterized by a kd of about 2 x 10’5to 3 x 10’5s’ 1 or less, a binding affinity for IL-4Ra characterized by an EC50 of about 0.02 to 1.5 nM or less, a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 7.3 nM or less, a blocking activity against IL- 13 Type II signaling characterized by an IC50 of about 3 to 3.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 8 to 50 nM or less.

78. The computer-implemented method of claim 69, wherein the predicted functional property is at least one of: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.02 to 0.04 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7.7 x 105to 8.9 x 105M_|s_|or less,a dissociation from IL-4Ra characterized by a kd of about 2.1 x 10'5to 2.6 x 10'5s'1or less, a binding affinity for IL-4Ra characterized by an ECso of about 0.04 to 1.3 nM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 4.4 to 7.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 30 to 50 nM or less.

79. The computer-implemented method of claim 69, wherein the predicted functional property is at least one of: a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 1.8 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 30 to 34 nM or less.

80. The computer-implemented method of claim 69, wherein the predicted functional property is an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 8 to 8.8 nM or less.

81. The computer-implemented method of claim 69, wherein the predicted functional property relates to modulating activity of a target molecule, and wherein the target molecule is interleukin-4 receptor alpha (IL-4Ra).

82. The computer-implemented method of claim 69, wherein the analysis score is a binding score.

83. The computer-implemented method of claim 69, wherein the predicted functional property is a binding affinity.

84. The computer-implemented method of claim 69, wherein the polypeptide does not comprise a heavy chain having an amino acid sequence that is identical to SEQ ID NO: 104 and a light chain having an amino acid sequence that is identical to SEQ ID NO: 161.

85. A computer-based system for predicting a functional property of a polypeptide, the computer-based system comprising: a processor; and a memory with computer code instructions stored thereon, the processor and the memory, with the computer code instructions, being configured to cause the computer-based system to: implement a computationally binding optimized (CBO) model, the CBO model configured to: for each amino acid position of an amino acid sequence of the polypeptide: determine a plurality of energy scores based on the amino acid position in the amino acid sequence; generate a partition function based on the plurality of energy scores determined; and determine a cross-entropy metric based on (i) an amino acid at the amino acid position in the amino acid sequence, (ii) a maximum energy score of the plurality of energy scores determined, and (iii) the generated partition function; and generate an analysis score of the polypeptide based on each cross-entropy metric determined, the analysis score indicating a predicted functional property of the polypeptide, wherein the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 37, and wherein the polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.

86. A polypeptide that binds interleukin-4 receptor alpha (IL-4Ra): wherein the polypeptide is assigned a score above a threshold by an analysis via a computationally binding optimized (CBO) model,wherein the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 37, and wherein the polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.

87. The polypeptide of claim 86, wherein at least one of: the analysis is implementable by a script of Appendix A or Appendix B employing the CBO model, the CBO model is calculated using a table substantially similar to that of Appendix C, the score assigned to the polypeptide is generated using Appendix C, the polypeptide is assigned the score by the script of Appendix A or Appendix B, and the score assigned to the polypeptide is at least about -27.8.

88. The polypeptide of claim 86, wherein the polypeptide has at least one property selected from: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 1 pM or less, a binding affinity for IL-4Ra characterized by a kaof about 100 x 105M^s'1or less, a dissociation from IL-4Ra characterized by a kd of about 100 x 10'5s'1or less, a binding affinity for IL-4Ra characterized by an ECso of about 1 pM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 1 pM or less, a blocking activity against IL- 13 Type II signaling characterized by an ICso of about 1 pM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 1 pM or less.

89. The polypeptide of claim 86, wherein the polypeptide has at least one property selected from: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.05 to 0.5 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7 x 105to 9 x 105T's-1or less, a dissociation from IL-4Ra characterized by a kd of about 2 x 10’5to 3 x 10’5s’ 1 or less, a binding affinity for IL-4Ra characterized by an ECso of about 0.02 to 1.5 nM or less, a blocking activity against IL-4 Type II signaling characterized by an ICso of about 1.1 to 7.3 nM or less, a blocking activity against IL- 13 Type II signaling characterized by an ICso of about 3 to 3.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an ICso of about 8 to 50 nM or less.

90. The polypeptide of claim 86, wherein the polypeptide has at least one property selected from: a binding affinity for interleukin-4 receptor alpha (IL-4Ra) characterized by a KD of about 0.02 to 0.04 nM or less, a binding affinity for IL-4Ra characterized by a kaof about 7.7 x 105to 8.9 x105M_|s_|or less, a dissociation from IL-4Ra characterized by a kd of about 2.1 x 10’5to 2.6 x 10’5s’1or less, a binding affinity for IL-4Ra characterized by an EC50 of about 0.04 to 1.3 nM or less, a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 4.4 to 7.3 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 30 to 50 nM or less.

91. The polypeptide of claim 86, wherein the polypeptide has at least one property selected from:a blocking activity against IL-4 Type II signaling characterized by an IC50 of about 1.1 to 1.8 nM or less, and an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 30 to 34 nM or less.

92. The polypeptide of claim 86, wherein the polypeptide has an inhibitory activity against CD23 expression in B cells characterized by an IC50 of about 8 to 8.8 nM or less.

93. The polypeptide of claim 86, wherein the threshold is a score from the CBO model of one or more of a reference polypeptide that includes a VH and VL pair selected from:SEQ ID NO:5 and SEQ ID NO:24 (Reference);SEQ ID NO:6 and SEQ ID NO:25 (AB-1);SEQ ID NO:7 and SEQ ID NO:26 (AB-2);SEQ ID NO:8 and SEQ ID NO:27 (AB-3);SEQ ID NO:9 and SEQ ID NO:28 (AB-4);SEQ ID NO: 10 and SEQ ID NO:29 (AB-5);SEQ ID NO: 11 and SEQ ID NO:30 (AB-6);SEQ ID NO: 12 and SEQ ID NO: 31 (AB-7);SEQ ID NO: 13 and SEQ ID NO:32 (AB-8);SEQ ID NO: 14 and SEQ ID NO:33 (AB-9);SEQ ID NO: 15 and SEQ ID NO:34 (AB-10);SEQ ID NO: 16 and SEQ ID NO:35 (AB-11);SEQ ID NO: 17 and SEQ ID NO:36 (AB- 12);SEQ ID NO: 18 and SEQ ID NO:37 (AB-13);SEQ ID NO: 19 and SEQ ID NO:38 (AB- 14c);SEQ ID NO:20 and SEQ ID NO:39 (AB-15c); or a combination of any of the foregoing.

94. The polypeptide of claim 86, wherein the polypeptide does not comprise a heavy chain having an amino acid sequence that is identical to SEQ ID NO: 104 and a light chain having an amino acid sequence that is identical to SEQ ID NO: 161.

5. A polypeptide that binds human interleukin-4 receptor alpha (IL-4Ra), wherein the polypeptide is selected by a method comprising: evaluating a plurality of candidate polypeptides using a computationally binding optimized (CBO) model by, for each candidate polypeptide of the plurality of candidate polypeptides: for each amino acid position of an amino acid sequence of the candidate polypeptide: determining a plurality of energy scores based on the amino acid position in the amino acid sequence; generating a partition function based on the plurality of energy scores determined; and determining a cross-entropy metric based on (i) an amino acid at the amino acid position in the amino acid sequence, (ii) a maximum energy score of the plurality of energy scores determined, and (iii) the generated partition function; and generating an analysis score of the candidate polypeptide based on each cross-entropy metric determined, the analysis score indicating a functional property of the polypeptide’s ability to bind to human IL-4Ra; and selecting a given candidate polypeptide from among the plurality of candidate polypeptides based a result of the evaluating, wherein the polypeptide comprises an immunoglobulin heavy chain variable domain (VH) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 18 and an immunoglobulin light chain variable domain (VL) comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 37, and wherein the polypeptide does not comprise a VH comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:5 and a VL comprising an amino acid sequence having 100% sequence identity to SEQ ID NO:24.

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