ANTI-IFNAR1 ANTIBODIES FOR TREATING AUTOIMMUNE DISEASES

Abstract

Provided are antibodies or fragments thereof having binding specificity to the human interferon alpha and beta receptor subunit 1 (IFNAR1) protein. In various examples, the antibodies or fragments thereof include a VH and VL CDRs as disclosed herein, or variants thereof. Methods of using the antibodies or fragments thereof for treating autoimmune diseases and disorders are also provided.

Description

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a divisional of U.S. application Ser. No. 16/963,128, filed Jul. 17, 2020, now U.S. Pat. No. 11,059,897, which is the U.S. National Stage Application under 35 U.S.C. 371 of International Application No. PCT/CN2019/106412, filed Sep. 18, 2019, which claims priority to International Application PCT/CN2018/106157, filed Sep. 18, 2018. The contents of each of the aforementioned are hereby incorporated by reference in their entirety into the present disclosure.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jul. 11, 2021, is named 271415US2_SL.txt and is 102,221 bytes in size.

BACKGROUND

[0003] Systemic lupus erythematosus (SLE), also known simply as lupus, is an autoimmune disease in which the body's immune system mistakenly attacks healthy tissue in many parts of the body. Common symptoms include painful and swollen joints, fever, chest pain, hair loss, mouth ulcers, swollen lymph nodes, feeling tired, and a red rash which is most commonly on the face. Often there are periods of illness, called flares, and periods of remission during which there are few symptoms.

[0004] The cause of SLE is not clear. The mechanism involves an immune response by autoantibodies against a person's own tissues. These are most commonly anti-nuclear antibodies and they result in inflammation. There is no cure for SLE. Treatments may include NSAIDs, corticosteroids, immunosuppressants, hydroxychloroquine, and methotrexate. SLE significantly increases the risk of cardiovascular disease with this being the most common cause of death.

[0005] Type I IFNs, particularly the IFN-.alpha.s and IFN-0, have received attention for their roles in the pathogenesis of SLE and other autoimmune and inflammatory syndromes. By signaling through a common receptor (IFNAR), these pleiotropic cytokines affect almost every aspect of innate and adaptive immune responses, including upregulation of MHC and costimulatory molecules, and production of B cell survival factors (BAFF, April) by antigen-presenting cells, culminating in the engagement and expansion of autoreactive T and B cells. Of particular relevance to lupus pathogenesis is the induction of type I IFNs under sterile conditions through the engagement of endosomal Toll-like receptors (TLRs) by self-nucleic acids. There has been extensive interest in creating treatments based on blocking reagents against either the multiple IFN-.alpha.s and the single IFN-.beta., or their common receptor.

SUMMARY

[0006] The present disclosure provides antibodies or fragments thereof having binding specificity to the human interferon alpha and beta receptor subunit 1 (IFNAR1) protein, as well as bispecific antibodies having specificity to IFNAR1 and another antigen such as BAFF. These antibodies and fragments are useful in the treatment of autoimmune diseases such as systemic lupus erythematosus.

[0007] One embodiment of the present disclosure provides an antibody or fragment thereof having specificity to a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein, wherein the antibody or fragment thereof comprises a heavy chain variable region comprising heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and a light chain variable region comprising light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are selected from the group consisting of: (a) HCDR1: DYYMH (SEQ ID NO: 77), HCDR2: RIDPEDGETKYAPKFQG (SEQ ID NO: 78) or RIDPEDAETKYAPKFQG (SEQ ID NO:79), HCDR3: GGNFYVMDY (SEQ ID NO: 80), LCDR1: KASQNVGTNVV (SEQ ID NO: 81), LCDR2: SASYRVS (SEQ ID NO: 82), and LCDR3: QQKNNYPYT (SEQ ID NO: 83); and (b) HCDR1: DYYIH (SEQ ID NO: 92), HCDR2: RIDPEDGETKYAPKFQG (SEQ ID NO: 93) or RIDPEDAETKYAPKFQG (SEQ ID NO:94), HCDR3: YHGYWALDY (SEQ ID NO: 95), LCDR1: KTSQNVGTNVA (SEQ ID NO: 96), LCDR2: STSYRYS (SEQ ID NO: 97), and LCDR3: HQYFSYPYT (SEQ ID NO: 98).

[0008] In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are HCDR1: DYYMH (SEQ ID NO: 77), HCDR2: RIDPEDAETKYAPKFQG (SEQ ID NO:79), HCDR3: GGNFYVMDY (SEQ ID NO: 80), LCDR1: KASQNVGTNVV (SEQ ID NO: 81), LCDR2: SASYRVS (SEQ ID NO: 82), and LCDR3: QQKNNYPYT (SEQ ID NO: 83).

[0009] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 7, and 84-87 (e.g., SEQ ID NO:85), or a peptide having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 7, and 84-87 (e.g., SEQ ID NO:85). In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 8, and 88-91 (e.g., SEQ ID NO:88), or a peptide having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 8, and 88-91 (e.g., SEQ ID NO:88). The heavy chain variable region and the light chain variable, as recited here, include the CDR regions as recited above.

[0010] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 85, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 88.

[0011] In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are HCDR1: DYYIH (SEQ ID NO: 92), HCDR2: RIDPEDAETKYAPKFQG (SEQ ID NO:94), HCDR3: YHGYWALDY (SEQ ID NO: 95), LCDR1: KTSQNVGTNVA (SEQ ID NO: 96), LCDR2: STSYRYS (SEQ ID NO: 97), and LCDR3: HQYFSYPYT (SEQ ID NO: 98).

[0012] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 73, and 99-102 (e.g., SEQ ID NO:100), or a peptide having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 73, and 99-102 (e.g., SEQ ID NO:100).

[0013] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 74, and 103-106 (e.g., SEQ ID NO:105), or a peptide having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 74, and 103-106 (e.g., SEQ ID NO:105).

[0014] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 100, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 105.

[0015] In another embodiments, provided is an antibody or fragment thereof having specificity to a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein, wherein the antibody or fragment thereof can bind to one or more amino acid residues selected from the group consisting of H273, L274, Y275, K276, and K278 of the IFNAR1 protein.

[0016] In some embodiments, the antibody or fragment thereof of claim can bind to at least two of the amino acid residues selected from the group, such as K276, and K278. In some embodiments, the antibody or fragment thereof of claim can bind to at least three of the amino acid residues selected from the group, such as H273, K276, and K278; L274, K276, and K278; Y275, K276, and K278; or Y275, K276, and K278. In some embodiments, the antibody or fragment thereof of claim can bind to at least four, five or all of the amino acid residues selected from the group.

[0017] Also provided, in one embodiment, is a bifunctional molecule, comprising a first antigen-binding portion having specificity to a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein and a second portion having specificity to a second protein, wherein the first antigen-binding portion comprises an antibody fragment of the present disclosure.

[0018] In some embodiments, the second portion comprises peptide edratide (hCDR1) or TACI-Ig. In some embodiments, the second portion is an antigen-binding fragment having specificity to a protein selected from the group consisting of BAFF, CD20, CD22, CTLA4, IL6, CXCL13 and C5. In some embodiments, the second portion is an antigen-binding fragment having specificity to a human B-cell-activating factor (BAFF) protein.

[0019] In some embodiments, the bifunctional molecule has a format comprising a full antibody fused to two single chain fragments (scFv) or to two Fab fragments (as illustrated as Format 1 herein). In some embodiments, the second portion comprises an antigen-binding fragment of Belimumab.

[0020] Uses and methods of the presently disclosed antibodies and fragments are also provided. For instance, the presently disclosed antibodies and fragments can be used for suppressing an immune response or treating an autoimmune disease or disorder in a patient in need thereof.

[0021] In some embodiments, the autoimmune disease or disorder is selected from the group consisting of type 1 diabetes, rheumatoid arthritis (RA), psoriasis/psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus (lupus), inflammatory bowel disease, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, and celiac disease. In some embodiments, the autoimmune disease or disorder is systemic lupus erythematosus.

BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIG. 1A-B show that the 8G11H and 485G10H antibodies efficiently blocked IFN.alpha.c2b-induced reporter gene expression, with potency comparable to that of the corresponding chimeric antibodies.

[0023] FIG. 2A-B show that 8G11H and 485G10H antibodies dose-dependently reversed IFN.alpha.c2b-mediated inhibition of Daudi cell proliferation.

[0024] FIG. 3 shows that 8G11H and 485G10H dose-dependently inhibited recombinant IFN.alpha.c2b-induced secretion of IP-10 by normal PBMC culture.

[0025] FIG. 4 shows that the 8G11H and 485G10H antibodies efficiently inhibited the in vitro differentiation of dendritic cells as demonstrated by significant reduction of differentiated cell surface markers CD38 and CD86 expression.

[0026] FIG. 5 shows that the 8G11H and 485G10H antibodies efficiently inhibited the in vitro differentiation of dendritic cells, which resulted in impaired responses in differentiated cell-mediated MLR as demonstrated by decreased production of IFN-.gamma. by CD4.sup.+ T cells.

[0027] FIG. 6 shows that humanized monoclonal antibodies 8G11H and 485G10H significantly inhibited the SLE plasma mediated dendritic cell development in in vitro system.

[0028] FIG. 7 depicts a schematic of designed four formats for the anti-IFNAR/BAFF bispecific antibody (Bi-BFINR).

[0029] FIG. 8 shows that the bispecific antibody exhibited similar activity with 8G11 antibody in binding to IFNAR-1 and comparable activity to Belimumab in binding to BAFF.

[0030] FIG. 9A-B show that Bi-BFINF antibody had comparable activity to chimeric 8G11 in blocking IFN.alpha.2b signaling (A); Bi-BFINF antibody exhibited better activity in blockade of BAFF-induced B cell proliferation (B).

[0031] FIG. 10A shows time-concentration profiles of 485G10H1L3 in 2 cynomolgus monkeys.

[0032] FIG. 10B shows time-concentration profiles of 8G11H2L1 in 2 cynomolgus monkeys.

[0033] FIG. 11A shows CD11C, CD38, CD86, MHC Class I, MHC Class II and IFN-alpha R1 expression by FACs after dosing at 24 h, 48 h and 72 h.

[0034] FIG. 11B shows the serum level of monkey neopterin, beta-2-microglobulin and CRP at 24 h, 48 h and 72 h after dosing.

[0035] FIG. 11C shows the expression levels of type I IFN-induced gene signatures in monkey PBMCs at 24 h, 48 h and 72 h after dosing.

DETAILED DESCRIPTION

Definitions

[0036] It is to be noted that the term "a" or "an" entity refers to one or more of that entity; for example, "an antibody," is understood to represent one or more antibodies. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0037] As used herein, an "antibody" or "antigen-binding polypeptide" refers to a polypeptide or a polypeptide complex that specifically recognizes and binds to an antigen. An antibody can be a whole antibody and any antigen binding fragment or a single chain thereof. Thus the term "antibody" includes any protein or peptide containing molecule that comprises at least a portion of an immunoglobulin molecule having biological activity of binding to the antigen. Examples of such include, but are not limited to a complementarity determining region (CDR) of a heavy or light chain or a ligand binding portion thereof, a heavy chain or light chain variable region, a heavy chain or light chain constant region, a framework (FR) region, or any portion thereof, or at least one portion of a binding protein.

[0038] The terms "antibody fragment" or "antigen-binding fragment", as used herein, is a portion of an antibody such as F(ab').sub.2, F(ab).sub.2, Fab', Fab, Fv, scFv and the like. Regardless of structure, an antibody fragment binds with the same antigen that is recognized by the intact antibody. The term "antibody fragment" includes aptamers, spiegelmers, and diabodies. The term "antibody fragment" also includes any synthetic or genetically engineered protein that acts like an antibody by binding to a specific antigen to form a complex.

[0039] A "single-chain variable fragment" or "scFv" refers to a fusion protein of the variable regions of the heavy (V.sub.H) and light chains (V.sub.L) of immunoglobulins. In some aspects, the regions are connected with a short linker peptide of ten to about 25 amino acids. The linker can be rich in glycine for flexibility, as well as serine or threonine for solubility, and can either connect the N-terminus of the V.sub.H with the C-terminus of the V.sub.L, or vice versa. This protein retains the specificity of the original immunoglobulin, despite removal of the constant regions and the introduction of the linker. ScFv molecules are known in the art and are described, e.g., in U.S. Pat. No. 5,892,019.

[0040] The term antibody encompasses various broad classes of polypeptides that can be distinguished biochemically. Those skilled in the art will appreciate that heavy chains are classified as gamma, mu, alpha, delta, or epsilon (.gamma., .mu., .alpha., .delta., ) with some subclasses among them (e.g., .gamma.1-.gamma.4). It is the nature of this chain that determines the "class" of the antibody as IgG, IgM, IgA IgG, or IgE, respectively. The immunoglobulin subclasses (isotypes) e.g., IgG.sub.1, IgG.sub.2, IgG.sub.3, IgG.sub.4, IgG.sub.5, etc. are well characterized and are known to confer functional specialization. Modified versions of each of these classes and isotypes are readily discernable to the skilled artisan in view of the instant disclosure and, accordingly, are within the scope of the instant disclosure. All immunoglobulin classes are clearly within the scope of the present disclosure, the following discussion will generally be directed to the IgG class of immunoglobulin molecules. With regard to IgG, a standard immunoglobulin molecule comprises two identical light chain polypeptides of molecular weight approximately 23,000 Daltons, and two identical heavy chain polypeptides of molecular weight 53,000-70,000. The four chains are typically joined by disulfide bonds in a "Y" configuration wherein the light chains bracket the heavy chains starting at the mouth of the "Y" and continuing through the variable region.

[0041] Antibodies, antigen-binding polypeptides, variants, or derivatives thereof of the disclosure include, but are not limited to, polyclonal, monoclonal, multispecific, human, humanized, primatized, or chimeric antibodies, single chain antibodies, epitope-binding fragments, e.g., Fab, Fab' and F(ab').sub.2, Fd, Fvs, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv), fragments comprising either a VK or VH domain, fragments produced by a Fab expression library, and anti-idiotypic (anti-Id) antibodies (including, e.g., anti-Id antibodies to LIGHT antibodies disclosed herein) Immunoglobulin or antibody molecules of the disclosure can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass of immunoglobulin molecule.

[0042] Light chains are classified as either kappa or lambda (K, .lamda.). Each heavy chain class may be bound with either a kappa or lambda light chain. In general, the light and heavy chains are covalently bonded to each other, and the "tail" portions of the two heavy chains are bonded to each other by covalent disulfide linkages or non-covalent linkages when the immunoglobulins are generated either by hybridomas, B cells or genetically engineered host cells. In the heavy chain, the amino acid sequences run from an N-terminus at the forked ends of the Y configuration to the C-terminus at the bottom of each chain.

[0043] Both the light and heavy chains are divided into regions of structural and functional homology. The terms "constant" and "variable" are used functionally. In this regard, it will be appreciated that the variable domains of both the light (VK) and heavy (VH) chain portions determine antigen recognition and specificity. Conversely, the constant domains of the light chain (CK) and the heavy chain (CH1, CH2 or CH3) confer important biological properties such as secretion, transplacental mobility, Fc receptor binding, complement binding, and the like. By convention the numbering of the constant region domains increases as they become more distal from the antigen-binding site or amino-terminus of the antibody. The N-terminal portion is a variable region and at the C-terminal portion is a constant region; the CH3 and CK domains actually comprise the carboxy-terminus of the heavy and light chain, respectively.

[0044] As indicated above, the variable region allows the antibody to selectively recognize and specifically bind epitopes on antigens. That is, the VK domain and VH domain, or subset of the complementarity determining regions (CDRs), of an antibody combine to form the variable region that defines a three dimensional antigen-binding site. This quaternary antibody structure forms the antigen-binding site present at the end of each arm of the Y. More specifically, the antigen-binding site is defined by three CDRs on each of the VH and VK chains (i.e. CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3). In some instances, e.g., certain immunoglobulin molecules derived from camelid species or engineered based on camelid immunoglobulins, a complete immunoglobulin molecule may consist of heavy chains only, with no light chains. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993).

[0045] In naturally occurring antibodies, the six "complementarity determining regions" or "CDRs" present in each antigen-binding domain are short, non-contiguous sequences of amino acids that are specifically positioned to form the antigen-binding domain as the antibody assumes its three dimensional configuration in an aqueous environment. The remainder of the amino acids in the antigen-binding domains, referred to as "framework" regions, show less inter-molecular variability. The framework regions largely adopt a .beta.-sheet conformation and the CDRs form loops which connect, and in some cases form part of, the .beta.-sheet structure. Thus, framework regions act to form a scaffold that provides for positioning the CDRs in correct orientation by inter-chain, non-covalent interactions. The antigen-binding domain formed by the positioned CDRs defines a surface complementary to the epitope on the immunoreactive antigen. This complementary surface promotes the non-covalent binding of the antibody to its cognate epitope. The amino acids comprising the CDRs and the framework regions, respectively, can be readily identified for any given heavy or light chain variable region by one of ordinary skill in the art, since they have been precisely defined (see "Sequences of Proteins of Immunological Interest," Kabat, E., et al., U.S. Department of Health and Human Services, (1983); and Chothia and Lesk, J. Mol. Biol., 196:901-917 (1987)).

[0046] In the case where there are two or more definitions of a term which is used and/or accepted within the art, the definition of the term as used herein is intended to include all such meanings unless explicitly stated to the contrary. A specific example is the use of the term "complementarity determining region" ("CDR") to describe the non-contiguous antigen combining sites found within the variable region of both heavy and light chain polypeptides. This particular region has been described by Kabat et al., U.S. Dept. of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983) and by Chothia et al., J. Mol. Biol. 196:901-917 (1987), which are incorporated herein by reference in their entireties. The CDR definitions according to Kabat and Chothia include overlapping or subsets of amino acid residues when compared against each other. Nevertheless, application of either definition to refer to a CDR of an antibody or variants thereof is intended to be within the scope of the term as defined and used herein. The appropriate amino acid residues which encompass the CDRs as defined by each of the above cited references are set forth in the table below as a comparison. The exact residue numbers which encompass a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in the art can routinely determine which residues comprise a particular CDR given the variable region amino acid sequence of the antibody.

[0047] Antibodies disclosed herein may be from any animal origin including birds and mammals Preferably, the antibodies are human, murine, donkey, rabbit, goat, guinea pig, camel, llama, horse, or chicken antibodies. In another embodiment, the variable region may be condricthoid in origin (e.g., from sharks).

[0048] As used herein, the term "heavy chain constant region" includes amino acid sequences derived from an immunoglobulin heavy chain. A polypeptide comprising a heavy chain constant region comprises at least one of: a CH1 domain, a hinge (e.g., upper, middle, and/or lower hinge region) domain, a CH2 domain, a CH3 domain, or a variant or fragment thereof. For example, an antigen-binding polypeptide for use in the disclosure may comprise a polypeptide chain comprising a CH1 domain; a polypeptide chain comprising a CH1 domain, at least a portion of a hinge domain, and a CH2 domain; a polypeptide chain comprising a CH1 domain and a CH3 domain; a polypeptide chain comprising a CH1 domain, at least a portion of a hinge domain, and a CH3 domain, or a polypeptide chain comprising a CH1 domain, at least a portion of a hinge domain, a CH2 domain, and a CH3 domain. In another embodiment, a polypeptide of the disclosure comprises a polypeptide chain comprising a CH3 domain. Further, an antibody for use in the disclosure may lack at least a portion of a CH2 domain (e.g., all or part of a CH2 domain). As set forth above, it will be understood by one of ordinary skill in the art that the heavy chain constant region may be modified such that they vary in amino acid sequence from the naturally occurring immunoglobulin molecule.

[0049] The heavy chain constant region of an antibody disclosed herein may be derived from different immunoglobulin molecules. For example, a heavy chain constant region of a polypeptide may comprise a CH1 domain derived from an IgG1 molecule and a hinge region derived from an IgG.sub.3 molecule. In another example, a heavy chain constant region can comprise a hinge region derived, in part, from an IgG.sub.1 molecule and, in part, from an IgG.sub.3 molecule. In another example, a heavy chain portion can comprise a chimeric hinge derived, in part, from an IgG.sub.1 molecule and, in part, from an IgG.sub.4 molecule.

[0050] As used herein, the term "light chain constant region" includes amino acid sequences derived from antibody light chain. Preferably, the light chain constant region comprises at least one of a constant kappa domain or constant lambda domain.

[0051] A "light chain-heavy chain pair" refers to the collection of a light chain and heavy chain that can form a dimer through a disulfide bond between the CL domain of the light chain and the CH1 domain of the heavy chain.

[0052] As previously indicated, the subunit structures and three dimensional configuration of the constant regions of the various immunoglobulin classes are well known. As used herein, the term "VH domain" includes the amino terminal variable domain of an immunoglobulin heavy chain and the term "CH1 domain" includes the first (most amino terminal) constant region domain of an immunoglobulin heavy chain. The CH1 domain is adjacent to the VH domain and is amino terminal to the hinge region of an immunoglobulin heavy chain molecule.

[0053] As used herein the term "CH2 domain" includes the portion of a heavy chain molecule that extends, e.g., from about residue 244 to residue 360 of an antibody using conventional numbering schemes (residues 244 to 360, Kabat numbering system; and residues 231-340, EU numbering system; see Kabat et al., U.S. Dept. of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983). The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It is also well documented that the CH3 domain extends from the CH2 domain to the C-terminal of the IgG molecule and comprises approximately 108 residues.

[0054] As used herein, the term "hinge region" includes the portion of a heavy chain molecule that joins the CH1 domain to the CH2 domain. This hinge region comprises approximately 25 residues and is flexible, thus allowing the two N-terminal antigen-binding regions to move independently. Hinge regions can be subdivided into three distinct domains: upper, middle, and lower hinge domains (Roux et al., J. Immunol 161:4083 (1998)).

[0055] As used herein the term "disulfide bond" includes the covalent bond formed between two sulfur atoms. The amino acid cysteine comprises a thiol group that can form a disulfide bond or bridge with a second thiol group. In most naturally occurring IgG molecules, the CH1 and CK regions are linked by a disulfide bond and the two heavy chains are linked by two disulfide bonds at positions corresponding to 239 and 242 using the Kabat numbering system (position 226 or 229, EU numbering system).

[0056] As used herein, the term "chimeric antibody" will be held to mean any antibody wherein the immunoreactive region or site is obtained or derived from a first species and the constant region (which may be intact, partial or modified in accordance with the instant disclosure) is obtained from a second species. In certain embodiments the target binding region or site will be from a non-human source (e.g. mouse or primate) and the constant region is human.

[0057] By "specifically binds" or "has specificity to," it is generally meant that an antibody binds to an epitope via its antigen-binding domain, and that the binding entails some complementarity between the antigen-binding domain and the epitope. According to this definition, an antibody is said to "specifically bind" to an epitope when it binds to that epitope, via its antigen-binding domain more readily than it would bind to a random, unrelated epitope. The term "specificity" is used herein to qualify the relative affinity by which a certain antibody binds to a certain epitope. For example, antibody "A" may be deemed to have a higher specificity for a given epitope than antibody "B," or antibody "A" may be said to bind to epitope "C" with a higher specificity than it has for related epitope "D."

[0058] As used herein, the terms "treat" or "treatment" refer to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, such as the progression of cancer. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "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.

[0059] By "subject" or "individual" or "animal" or "patient" or "mammal," is meant any subject, particularly a mammalian subject, for whom diagnosis, prognosis, or therapy is desired. Mammalian subjects include humans, domestic animals, farm animals, and zoo, sport, or pet animals such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, cows, and so on.

[0060] As used herein, phrases such as "to a patient in need of treatment" or "a subject in need of treatment" includes subjects, such as mammalian subjects, that would benefit from administration of an antibody or composition of the present disclosure used, e.g., for detection, for a diagnostic procedure and/or for treatment.

Anti-IFNAR1 Antibodies

[0061] The present disclosure provides antibodies, including bispecific antibodies and fragments, that have binding specificity to the human interferon alpha and beta receptor subunit 1 (IFNAR1) protein. As demonstrated in the experimental examples, numerous murine anti-human IFNAR1 antibodies were obtained, having high binding affinity to the human IFNAR1 protein. Two of the murine antibody clones, 8G11 and 485G10, were selected for further humanization and characterization. The humanized antibodies dose-dependently reversed IFN.alpha.c2b-mediated inhibition of Daudi cell proliferation, IFN.alpha.c2b-induced anti-viral function as well as efficiently inhibited the cell signal elicited by multiple type I IFNs at pM levels. The IFNs included IFN.alpha. 1, IFN.alpha. 2a, IFN.alpha. 4, IFN.alpha. 5, IFN.alpha. 6, IFN.alpha. 7, IFN.alpha. 8, IFN.alpha. 10, IFN.alpha. 14, IFN.alpha. 16, IFN.alpha. 17, and IFN.alpha. 21.

[0062] Additional functional studies showed that these antibodies efficiently inhibited IFN.alpha.2b-induced secretion of IP-10 by human PBMCs and inhibited IFN.alpha.-dependent or SLE plasma-mediated dendritic cell development as demonstrated by reduced cell surface markers CD38 and CD86 expression, and decreased production of IFN-.gamma. by CD4+ T cells in MLR system.

[0063] Various formats of anti-IFNAR1/anti-BAFF bispecific antibodies were also prepared and tested for their binding capabilities and biological functions. These bispecific antibodies exhibited comparable binding activity to monospecific 8G11 in blocking IFN.alpha.2b signaling, and even better activity in blockade of BAFF-induced B cell proliferation.

[0064] In accordance with one embodiment of the present disclosure, provided are antibodies and fragments thereof that include the heavy chain and light chain variable domains with the CDR regions of the antibodies prepared in the experimental examples. The CDRs are summarized in Table A below.

TABLE-US-00001 TABLE A CDR Sequences Antibody CDR Sequences (CDR1, CDR2, chain CDR3 in order, for VH or VL) SEQ ID NO: 4A6-VH DYYMH 77 RIDPDDGETKYAPKFQG 107 GGNYYVMDN 108 4A6-VL KASQNVGTNVA 109 TASYRYS 110 QQYFSYPHT 111 4B12-VH DSYMH 112 RIDPEDGETNYAPKFQG 113 RVSSLYAMDY 114 4B12-VL KASQNVGTNVA 109 LASYRYS 115 QQYNNYPWT 116 4D8-VH DYYMH 77 RIDPEDAETKYAPKFQG 79 GGNFYVMDY 80 4D8-VL KASQNVGTNVV 81 SASYRYS 117 QQKNSYPYT 118 8G11-VH DYYMH 77 RIDPEDGETKYAPKFQG 78 GGNFYVMDY 80 8G11-VL KASQNVGTNVV 81 SASYRVS 82 QQKNNYPYT 83 12G11-VH DYYMH 77 RIDPEDGETKYAPKFQG 78 GGNYYAMDY 119 12G11-VL KASQNVGTNVA 109 SASYRYS 117 QQHNSYTYK 120 17F9-VH DYYIH 92 RIDPEDGETKYAPKFQD 121 YDGYYGFDY 122 17F9-VL KASQNVGTNVA 109 STSYRYS 97 HQYNNYPYT 123 18B6-VH DYYMC 124 RIDPEDGETKYAPKFQG 78 GGNYYAMDY 119 18B6-VL KASQNVGTNVA 109 SATYRYS 125 QQHNSYSYT 126 18C1-VH DYYMH 77 RIDPEDGETKYAPKFQG 78 LGNWVFDY 127 18C1-VL KASQNVGTNVD 128 SASYRYS 117 QQYNTYT 129 19B6-VH DYYMH 77 RIDPEDGETKYAPKFQV 130 GGNFYYFDY 131 19B6-VL KASQNVGTNVA 109 SASYRYS 117 QQCINYPYT 132 20B3-VH DYYIH 92 RIDPEDGETKYAPTFQG 133 YNGYSGFDY 134 20B3-VL KASQNVGTNVV 81 SASYRYS 117 QQYNRYPFT 135 20E10-VH DYYIH 92 RIDPEDGETKYAPKFQD 121 YDGYYGFDY 122 20E10-VL KASQNVGTNVA 109 STSYRYS 97 HQYNNYPYT 123 20E12-VH DYYMH 77 RIDPEDGETKYVPKFQG 136 GGSYYVMDY 137 20E12-VL KASQNVGTSVA 138 SASYRYS 117 QQDNSYPHT 139 21D6-VH DYYMH 77 RIDPEDGETKYAPKFQG 78 LHWSLDS 140 21D6-VL KASQNVGTAVA 141 STANRDT 142 QQYSSYPYT 143 24F6-VH DYYIH 92 RVDPEDGETKYVPKFLD 144 GGNYYAMDY 119 24F6-VL KASQNVGTNVA 109 LASYRYS 115 QQCNNYRLT 145 29E12-VH DYYIH 92 RIDPEDGETKYAPKFQD 121 YDGYYGFDY 122 29E12-VL KASQNVGTNVA 109 STSYRYS 97 HQYNNYPYT 123 30B5-VH DSYIH 146 RIDPEDGETKYAPKFQG 78 WLADYSAMDN 147 30B5-VL KASEDIYNRLA 148 GATSLET 149 QQYWNTLYT 150 30C8-VH DSYMH 112 RIDPEDGETKYAPKFQG 78 RGSSLYAVDY 151 30C8-VL KASQNVGTSVA 138 LASYRHR 152 QQFNIYPWT 153 34H8-VH DYYLH 154 RIDPEDGETKYAPKFQG 78 GGNYDVMDY 155 34H8-VL KASQNVGTYVV 156 SASYRYS 117 QQKNTYPFT 157 36E3-VH DSYMH 112 RIDPEDGETKYAPKFQG 78 RGSSLYAVDY 151 36E3-VL KASQNVGTSVA 138 LASYRHR 152 QQFNIYPWT 153 39F5-VH DYYIH 92 RIDPEDGETKYAPKFQV 130 GGNFYYFDY 131 39F5-VL KASQNVGTNVA 109 SASYRYS 117 QQCINYPYT 132 41F1-VH DYYMH 77 RIDPEDGETKYVPKFQG 136 GGNYYVMDY 158 41F1-VL KASQNVGTYVA 159 SASYRYN 160 QQYNNYPLT 161 46A10-VH DYYMH 77 RIDPEDGETKYAPKFQV 130 GGNFYYFDF 162 46A10-VL KASQNVGTNVA 109 SASYRYS 117 QQCINYPYT 132 47C6-VH DYYIH 92 RIDPEDGETKYAPKFQG 78 GGNFYYFDY 131 47C6-VL KASQNVGTNVA 109 SASYRYS 117 QQCNSYSYT 163 47F5-VH DYYMH 77 RIDPEDGETKYVPKFQG 136 GGSYYVMDY 137 47F5-VL KASQNVGTSVA 138 SASYRYS 117 QQDNSYPHT 139 104A3-VH DYYIH 92 RIDPEDGETKYAPKFQG 78 YDGYYCFDY 164 104A3-VL KASQNVGTNVA 109 STSYRYS 97 QQYNNYPYT 165 106A7-VH DYYMH 77 RIDPEDGETKYAPKFQG 78 DWGHSFDY 166 106A7-VL KASQNVGTTVA 167 SASYRYS 117 QQYNSYT 168 107E12-VH DYYIH 92 RIDPEDGETKYAPKFQD 121 EGSFTGWFPY 169 107E12-VL KARQSVGTYVA 170 STSYRYN 171 QQYHSYPYT 172 124D12-VH DYYIH 92 RIDPEDGETKYAPKFQG 78 YDGYYCFDY 164 124D12-VL KASQNVGTNVA 109 STSYRYS 97 QQYNNYPYT 165 260H8-VH SDYWN 173 YISYSGSIYYNPSLKS 174 SGGMYYFDY 175 260H8-VL RASGNIHNYLA 176 NAKTLED 177 QHFWSIPPT 178 268E9-VH SDYWN 173 YISYSGSIYYNPSLKS 174 SGGMYYFDY 175 268E9-VL RASGNIHNYLA 176 NAKTLED 177 QHFWSIPPT 178 269A7-VH SDYWN 173 YISYSGTIYYNPSLKS 179 SGGMYYFDY 175

269A7-VL RASGNIHNYLA 176 NAKTLED 177 QHFWSIPPT 178 293H10-VH SDYWN 173 YISYSGNTDYNPSLKS 180 SEGMYFFDY 181 293H10-VL RASGNIHNYLA 176 NAKTLAD 182 QHFWSTPPT 183 370E5-VH DYYIH 92 RIDPEDGETKYAPKFQG 78 FGGLTAMDY 184 370E5-VL KASQNVGTNVA 109 ATSYRYS 185 QQYNNYPYT 165 392D6-VH DYYVH 186 RIDPEDGETKYAPKFQG 78 FGGLDAMDY 187 392D6-VL KASQNVGTNVA 109 STSYRYN 171 QQFNRYPYT 188 402G3-VH SHFIH 189 WIYPGDDDTEYNHKFNG 190 RVEYYNGGFAY 191 402G3-VL KASKNIRNNLG 192 SGSTLQS 193 QQYDQYPLT 194 430H6-VH TYGMGVG 195 NIWWDDDKYYNPSLKN 196 HPLPGYKDNYVVDA 197 430H6-VL RSSQSLEYSDQYTYLE 198 GVSNRFS 199 FQATHDPYT 200 485G10-VH DYYIH 92 RIDPEDGETKYAPKFQG 78 YHGYWALDY 95 485G10-VL KTSQNVGTNVA 96 STSYRYS 97 HQYFSYPYT 98 487E3-VH DCYIH 201 RIDPEDGETKYAPKFQA 202 HCNFLYFDY 203 487E3-VL KASQNVGTIVA 204 SASYRSS 205 QQYNNYPVI 206

[0065] In some embodiments, the VH CDR1, CDR2, and CDR3 are selected from any set of VH CDR1, CDR2, and CDR3 shown in Table A, and the VL CDR1, CDR2, and CDR3 are selected from any set of VL CDR1, CDR2, and CDR3 shown in Table A. In some embodiments, the VH CDR1, CDR2, and CDR3 and the VL CDR1, CDR2, and CDR3 are selected from those derived from the same antibody in the examples.

[0066] In some embodiments, at least one, or two, or three, or four, or five, or six of the VH CDR1, CDR2, and CDR3 and the VL CDR1, CDR2, and CDR3 of the above are modified by one, two or three amino acid additions, deletions, substitutions, or the combinations thereof.

[0067] In one embodiment, the anti-IFNAR1 antibody or fragment thereof includes the following CDRs: HCDR1: DYYMH (SEQ ID NO: 77), HCDR2: RIDPEDGETKYAPKFQG (SEQ ID NO: 78), HCDR3: GGNFYVMDY (SEQ ID NO: 80), LCDR1: KASQNVGTNVV (SEQ ID NO: 81), LCDR2: SASYRVS (SEQ ID NO: 82), and LCDR3: QQKNNYPYT (SEQ ID NO: 83).

[0068] In one embodiment, one or more of the amino acid residues in the CDRs are substituted with a different amino acid to avoid post-translational modification. An example anti-IFNAR1 antibody or fragment thereof includes the following CDRs: HCDR1: DYYMH (SEQ ID NO: 77), HCDR2: RIDPEDAETKYAPKFQG (SEQ ID NO: 79), HCDR3: GGNFYVMDY (SEQ ID NO: 80), LCDR1: KASQNVGTNVV (SEQ ID NO: 81), LCDR2: SASYRVS (SEQ ID NO: 82), and LCDR3: QQKNNYPYT (SEQ ID NO: 83).

[0069] In some embodiments, the antibody is humanized but with one or more of the following back mutations on the heavy chain: 12V, 20L, 24G, 38K, 48I, 68A, 70I, 72A, 79A and 81L, according to Kabat numbering, and combinations thereof. In some embodiments, the antibody is humanized but with one or more of the following back mutations on the light chain: 4M, 13T, 21V, 43S, 46V, 74I, 78V and 87F according to Kabat numbering, and combinations thereof.

[0070] Non-limiting examples of heavy chain variable regions include SEQ ID NO: 7, and 84-87. Non-limiting example of light chain variable regions include SEQ ID NO: 8, and 88-91.

[0071] In some embodiments, the heavy chain variable region includes SEQ ID NO:85. In some embodiments, the light chain variable region includes SEQ ID NO:88.

[0072] In one embodiment, the anti-IFNAR1 antibody or fragment thereof includes the following CDRs: HCDR1: DYYIH (SEQ ID NO: 92), HCDR2: RIDPEDGETKYAPKFQG (SEQ ID NO: 93), HCDR3: YHGYWALDY (SEQ ID NO: 95), LCDR1: KTSQNVGTNVA (SEQ ID NO: 96), LCDR2: STSYRYS (SEQ ID NO: 97), and LCDR3: HQYFSYPYT (SEQ ID NO: C6).

[0073] In one embodiment, the anti-IFNAR1 antibody or fragment thereof includes the following CDRs: HCDR1: DYYIH (SEQ ID NO: 92), HCDR2: RIDPEDAETKYAPKFQG (SEQ ID NO:94), HCDR3: YHGYWALDY (SEQ ID NO: 95), LCDR1: KTSQNVGTNVA (SEQ ID NO: 96), LCDR2: STSYRYS (SEQ ID NO: 97), and LCDR3: HQYFSYPYT (SEQ ID NO: C6).

[0074] In some embodiments, the antibody is humanized but with one or more of the following back mutations on the heavy chain: 20L, 24G, 38K, 48I, 68A, 70I, 72A, 81L, and 97G, according to Kabat numbering, and combinations thereof. In some embodiments, the antibody is humanized but with one or more of the following back mutations on the light chain: 13T, 21V, 36Y, 46P, 78V, and 104L according to Kabat numbering, and combinations thereof.

[0075] Non-limiting examples of heavy chain variable regions include SEQ ID NO: 73, and 99-102. Non-limiting example of light chain variable regions include SEQ ID NO: 74, and 103-106.

[0076] In some embodiments, the heavy chain variable region includes SEQ ID NO:100. In some embodiments, the light chain variable region includes SEQ ID NO:105.

[0077] It was an interesting discovery that the presently prepared antibodies target an epitope that is different from known anti-IFNAR1 antibody Anifrolumab. Accordingly, in one embodiment, provided is an antibody or fragment thereof having specificity to a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein, wherein the antibody or fragment thereof can bind to one or more amino acid residues selected from the group consisting of H273, L274, Y275, K276, and K278 of the IFNAR1 protein.

[0078] In some embodiments, the antibody or fragment can bind to at least two of these epitope residues, such as H273 and L274, H273 and Y275, H273 and K276, H273 and K278, L274 and Y275, L274 and K276, L274 and K278, Y275 and K276, Y275 and K278, or K276 and K278.

[0079] In some embodiments, the antibody or fragment can bind to at least two of these epitope residues, such as H273, L274, and Y275; H273, L274, and K276; H273, L274, and K278; H273, Y275, and K276; H273, Y275, and K278; H273, K276, and K278; L274, Y275, and K276; L274, Y275, and K278; L274, K276, and K278; and Y275, K276, and K278.

[0080] In some embodiments, the antibody or fragment can bind to at least four of these epitope residues, such as L274, Y275, K276, and K278; H273, Y275, K276, and K278; H273, L274, K276, and K278; H273, L274, Y275, and K278; and H273, L274, Y275, and K276.

[0081] In some embodiments, the antibody or fragment can bind to all of these epitope residues.

[0082] The CDRs, heavy chain variable regions and light chain variable regions of the present disclosure can be further modified. In some embodiments, the modified heavy chain variable region or light chain variable region retains at least about 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% sequence identity and is still capable of binding to IFNAR1.

[0083] In some embodiments, the modification is substitution at no more than one hot spot position from each of the CDRs. In some embodiments, the modification is substitution at one, two or three such hot spot positions. In one embodiment, the modification is substitution at one of the hot spot positions. Such substitutions, in some embodiments, are conservative substitutions.

[0084] A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a nonessential amino acid residue in an immunoglobulin polypeptide is preferably replaced with another amino acid residue from the same side chain family. In another embodiment, a string of amino acids can be replaced with a structurally similar string that differs in order and/or composition of side chain family members.

[0085] Non-limiting examples of conservative amino acid substitutions are provided in the table below, where a similarity score of 0 or higher indicates conservative substitution between the two amino acids.

Amino Acid Similarity Matrix

TABLE-US-00002

[0086] C G P S A T D E N Q H K R V M I L F Y W W -8 -7 -6 -2 -6 -5 -7 -7 -4 -5 -3 -3 2 -6 -4 -5 -2 0 0 17 Y 0 -5 -5 -3 -3 -3 -4 -4 -2 -4 0 -4 -5 -2 -2 -1 -1 7 10 F -4 -5 -5 -3 -4 -3 -6 -5 -4 -5 -2 -5 -4 -1 0 1 2 9 L -6 -4 -3 -3 -2 -2 -4 -3 -3 -2 -2 -3 -3 2 4 2 6 I -2 -3 -2 -1 -1 0 -2 -2 -2 -2 -2 -2 -2 4 2 5 M -5 -3 -2 -2 -1 -1 -3 -2 0 -1 -2 0 0 2 6 V -2 -1 -1 -1 0 0 -2 -2 -2 -2 -2 -2 -2 4 R -4 -3 0 0 -2 -1 -1 -1 0 1 2 3 6 K -5 -2 -1 0 -1 0 0 0 1 1 0 5 H -3 -2 0 -1 -1 -1 1 1 2 3 6 Q -5 -1 0 -1 0 -1 2 2 1 4 N -4 0 -1 1 0 0 2 1 2 E -5 0 -1 0 0 0 3 4 D -5 1 -1 0 0 0 4 T -2 0 0 1 1 3 A -2 1 1 1 2 S 0 1 1 1 P -3 -1 6 G -3 5 C 12

Conservative Amino Acid Substitutions

TABLE-US-00003

[0087] For Amino Acid Substitution With Alanine D-Ala, Gly, Aib, .beta.-Ala, L-Cys, D-Cys Arginine D-Arg, Lys, D-Lys, Orn D-Orn Asparagine D-Asn, Asp, D-Asp, Glu, D-Glu Gln, D-Gln Aspartic Acid D-Asp, D-Asn, Asn, Glu, D-Glu, Gln, D-Gln Cysteine D-Cys, S-Me-Cys, Met, D-Met, Thr, D-Thr, L-Ser, D-Ser Glutamine D-Gln, Asn, D-Asn, Glu, D-Glu, Asp, D-Asp Glutamic Acid D-Glu, D-Asp, Asp, Asn, D-Asn, Gln, D-Gln Glycine Ala, D-Ala, Pro, D-Pro, Aib, .beta.-Ala Isoleucine D-Ile, Val, D-Val, Leu, D-Leu, Met, D-Met Leucine Val, D-Val, Met, D-Met, D-Ile, D-Leu, Ile Lysine D-Lys, Arg, D-Arg, Orn, D-Orn Methionine D-Met, S-Me-Cys, Ile, D-Ile, Leu, D-Leu, Val, D-Val Phenylalanine D-Phe, Tyr, D-Tyr, His, D-His, Trp, D-Trp Proline D-Pro Serine D-Ser, Thr, D-Thr, allo-Thr, L-Cys, D-Cys Threonine D-Thr, Ser, D-Ser, allo-Thr, Met, D-Met, Val, D-Val Tyrosine D-Tyr, Phe, D-Phe, His, D-His, Trp, D-Trp Valine D-Val, Leu, D-Leu, Ile, D-Ile, Met, D-Met

[0088] It will also be understood by one of ordinary skill in the art that antibodies as disclosed herein may be modified such that they vary in amino acid sequence from the naturally occurring binding polypeptide from which they were derived. For example, a polypeptide or amino acid sequence derived from a designated protein may be similar, e.g., have a certain percent identity to the starting sequence, e.g., it may be 60%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to the starting sequence.

[0089] In certain embodiments, the antibody comprises an amino acid sequence or one or more moieties not normally associated with an antibody. Exemplary modifications are described in more detail below. For example, an antibody of the disclosure may comprise a flexible linker sequence, or may be modified to add a functional moiety (e.g., PEG, a drug, a toxin, or a label).

[0090] Antibodies, variants, or derivatives thereof of the disclosure include derivatives that are modified, i.e., by the covalent attachment of any type of molecule to the antibody such that covalent attachment does not prevent the antibody from binding to the epitope. For example, but not by way of limitation, the antibodies can be modified, e.g., by glycosylation, acetylation, pegylation, phosphorylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. Any of numerous chemical modifications may be carried out by known techniques, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Additionally, the antibodies may contain one or more non-classical amino acids.

[0091] In some embodiments, the antibodies may be conjugated to therapeutic agents, prodrugs, peptides, proteins, enzymes, viruses, lipids, biological response modifiers, pharmaceutical agents, or PEG.

[0092] The antibodies may be conjugated or fused to a therapeutic agent, which may include detectable labels such as radioactive labels, an immunomodulator, a hormone, an enzyme, an oligonucleotide, a photoactive therapeutic or diagnostic agent, a cytotoxic agent, which may be a drug or a toxin, an ultrasound enhancing agent, a non-radioactive label, a combination thereof and other such agents known in the art.

Bi-Functional Molecules

[0093] It is contemplated that binding of both the IFNAR1 and another protein involved in the immune system would be advantageous or even synergistic. Accordingly, in some embodiments, bi-functional molecules are provided, such as a bispecific antibody with an anti-IFNAR1 fragment as disclosed herein and another antigen-binding fragment/portion.

[0094] The second portion of the bi-functional molecule may be any one of the following, (1) targeting B cells, including targeting B cell growth and survival factor such as anti-BAFF fragment, TACI-Ig; targeting surface molecules of B cells, such as anti-CD20 and anti-CD22 antibody fragments; (2) targeting co-stimulatory molecules, such as CTLA4-Ig and anti-CTLA4 antibody fragment; (3) targeting T cells, such as Edratide (hCDR1); and (4) targeting cytokines and complements, such anti-IL6, anti-CXCL13 and anti-05 antibody fragments.

[0095] In particular it is contemplated that the IFNAR1 and the BAFF (B-cell-activating factor) pathways can have synergistic effect in treating autoimmune diseases and disorders, such as lupus. Various formats of anti-IFNAR1/anti-BAFF bispecific antibodies (illustrated in FIG. 7) were tested for their binding capabilities and biological functions. Surprisingly, these bispecific antibodies exhibited comparable binding activity to monospecific 8G11 in blocking IFN.alpha.2b signaling, and even better activity in blockade of BAFF-induced B cell proliferation.

[0096] In one embodiment, therefore, provided is a bi-functional molecule having a first antigen-binding portion having specificity to a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein and a second antigen-binding portion having specificity to a human B-cell-activating factor (BAFF) protein. The anti-IFNAR1 portion can a fragment of any embodiment of the present disclosure. The anti-BAFF portion can be a fragment from Belimumab (see, e.g., Dubey A K, et al., Journal of Pharmacology & Pharmacotherapeutics. 2011; 2(4):317-319).

[0097] The bi-functional molecule can take a format as illustrated in FIG. 7, e.g., having a full antibody fused to two single chain fragments (scFv) or to two Fab fragments. Other formats of bi-functional or bispecific molecules are also provided. In some embodiments, each of the anti-IFNAR1 and anti-BAFF fragments is independently selected from a Fab fragment, a single-chain variable fragment (scFv), or a single-domain antibody. In some embodiments, the bispecific antibody further includes a Fc fragment.

Polynucleotides Encoding the Antibodies and Methods of Preparing the Antibodies

[0098] The present disclosure also provides isolated polynucleotides or nucleic acid molecules encoding the antibodies, variants or derivatives thereof of the disclosure. The polynucleotides of the present disclosure may encode the entire heavy and light chain variable regions of the antigen-binding polypeptides, variants or derivatives thereof on the same polynucleotide molecule or on separate polynucleotide molecules. Additionally, the polynucleotides of the present disclosure may encode portions of the heavy and light chain variable regions of the antigen-binding polypeptides, variants or derivatives thereof on the same polynucleotide molecule or on separate polynucleotide molecules.

[0099] Methods of making antibodies are well known in the art and described herein. In certain embodiments, both the variable and constant regions of the antigen-binding polypeptides of the present disclosure are fully human Fully human antibodies can be made using techniques described in the art and as described herein. For example, fully human antibodies against a specific antigen can be prepared by administering the antigen to a transgenic animal which has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled. Exemplary techniques that can be used to make such antibodies are described in U.S. Pat. Nos. 6,150,584; 6,458,592; 6,420,140 which are incorporated by reference in their entireties.

Treatment and Diagnostic Methods

[0100] As described herein, the antibodies, variants or derivatives of the present disclosure may be used in certain treatment and diagnostic methods.

[0101] The present disclosure is further directed to antibody-based therapies which involve administering the antibodies of the disclosure to a patient such as an animal, a mammal, and a human for treating one or more of the disorders or conditions described herein. Therapeutic compounds of the disclosure include, but are not limited to, antibodies of the disclosure (including variants and derivatives thereof as described herein) and nucleic acids or polynucleotides encoding antibodies of the disclosure (including variants and derivatives thereof as described herein).

[0102] One embodiment provides a method of suppressing an immune response in a patient in need thereof. The method entails administering to the patient an antibody, fragment, or bi-functional molecule of the present disclosure. In some embodiments, the patient is a tissue or organ transplant recipient.

[0103] In some embodiments, a method of treating an autoimmune disease or disorder is provided. Non-limiting examples of autoimmune disease or disorder include type 1 diabetes, rheumatoid arthritis (RA), psoriasis/psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus (lupus), inflammatory bowel disease, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, and celiac disease.

[0104] In a particular embodiments, the method is useful for treating systemic lupus erythematosus (lupus).

[0105] A specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the particular antibodies, variant or derivative thereof used, the patient's age, body weight, general health, sex, and diet, and the time of administration, rate of excretion, drug combination, and the severity of the particular disease being treated. Judgment of such factors by medical caregivers is within the ordinary skill in the art. The amount will also depend on the individual patient to be treated, the route of administration, the type of formulation, the characteristics of the compound used, the severity of the disease, and the desired effect. The amount used can be determined by pharmacological and pharmacokinetic principles well known in the art.

[0106] Methods of administration of the antibodies, variants or include but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The antigen-binding polypeptides or compositions may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Thus, pharmaceutical compositions containing the antigen-binding polypeptides of the disclosure may be administered orally, rectally, parenterally, intracistemally, intravaginally, intraperitoneally, topically (as by powders, ointments, drops or transdermal patch), bucally, or as an oral or nasal spray.

[0107] The term "parenteral" as used herein refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intra-articular injection and infusion.

[0108] Administration can be systemic or local. In addition, it may be desirable to introduce the antibodies of the disclosure into the central nervous system by any suitable route, including intraventricular and intrathecal injection; intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir, such as an Ommaya reservoir. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent.

[0109] It may be desirable to administer the antigen-binding polypeptides or compositions of the disclosure locally to the area in need of treatment; this may be achieved by, for example, and not by way of limitation, local infusion during surgery, topical application, e.g., in conjunction, with a wound dressing after surgery, by injection, by means of a catheter, by means of a suppository, or by means of an implant, said implant being of a porous, non-porous, or gelatinous material, including membranes, such as sialastic membranes, or fibers. Preferably, when administering a protein, including an antibody, of the disclosure, care must be taken to use materials to which the protein does not absorb.

[0110] Methods of detecting expression of a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein in a sample are also provided, in some embodiments, comprising contacting the sample with the antibody or fragment thereof, and detecting the binding which indicates expression of IFNAR1 in the sample.

Compositions

[0111] The present disclosure also provides pharmaceutical compositions. Such compositions comprise an effective amount of an antibody, and an acceptable carrier. In some embodiments, the composition further includes a second anticancer agent (e.g., an immune checkpoint inhibitor).

[0112] In a specific embodiment, the term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. Further, a "pharmaceutically acceptable carrier" will generally be a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

[0113] The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile 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. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates or phosphates. Antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; and agents for the adjustment of tonicity such as sodium chloride or dextrose are also envisioned. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences by E. W. Martin, incorporated herein by reference. Such compositions will contain a therapeutically effective amount of the antigen-binding polypeptide, preferably in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration. The parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0114] In an embodiment, the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to human beings. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.

[0115] The compounds of the disclosure can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

EXAMPLES

Example 1. Generation of Mouse Monoclonal Antibodies Against Human IFNAR-1

[0116] This example shows generation of anti-human-IFNAR-1 mouse monoclonal antibodies using the hybridoma technology.

Immunization

[0117] Recombinant human IFNAR-1 proteins containing the entire extracellular region of human IFNAR-1 were used as the immunogen to raise anti-human IFNAR-1 antibodies. C57BL/6, Balb/c, SJL mice or SD rats were first immunized subcutaneously (s.c.) with 50 .mu.g immunogen and then immunized intraperitoneally (i.p.). or s.c. biweekly with 25 .mu.g immunogen. Immune response was monitored by retroorbital bleeds. Plasma was screened by ELISA binding assay. In short, His-tagged IFNAR-1 was coated at 0.5 .mu.g/ml overnight and then blocked by 5% BSA in PBS. Serial diluted sera were incubated with the coated antigen for 1 h at room temperature. The resulting plates were washed with PBS/T and incubated with goat anti-mouse IgG-HRP for 1 h at room temperature. The plates were developed with TMB substrate and analyzed by spectrophotometer at OD 450-630 nm. The mice with high titers of anti-IFNAR1 immunoglobulin were selected for fusion and further screening. Three days prior to sacrifice and removal of the spleens, the mice were final boosted i.p. with 25 .mu.g antigen. The spleens were used for fusion.

Fusion and Hybridoma Screening

[0118] The mouse splenocytes, isolated from the mice, were fused with a mouse myeloma cell line based upon standard protocols. Single cell suspensions of splenic lymphocytes from immunized mice were fused to one-third the number of SP2/0 non secreting mouse myeloma cells with electrofusion machine. Cells were plated at approximately 1*10E5/well in flat bottom microtiter plate, followed by about 10 days incubation in selective medium containing 1*HAT, 10% fetal bovine serum, in DMEM. After 10 days, cells were cultured in medium in which the HAT was replaced with HT. Individual wells were then screened by ELISA for mouse anti-IFNAR-1 monoclonal IgG antibodies. The antibody secreting hybridomas were changed medium and screened again after 2 days. If still positive for mouse anti-IFNAR-1 antibodies, hybridoma were sub cloned twice by limiting dilution. The stable sub clones were then cultured in vitro to generate small amount of antibody in tissue culture medium for further characterization with various functional assays.

[0119] Clones showing strong blocking ability in IFN-responsive reporter assay were selected for sub cloning. Supernatants of 2-round sub clone were used to confirm ELISA-based human and rhesus IFNAR-1 binding and IFN alpha blocking ability, followed by sequencing and further analysis. After these screenings, 38 clones (4A6, 4B12, 4D8, 8G11, 12G11, 17F9, 18B6, 18C1, 19B6, 20B3, 20E10, 20E12, 21D6, 24F6, 29E12, 30B5, 3008, 34H8, 36E3, 39F5, 41F1, 46A10, 47C6, 47F5, 104A3, 106A7, 107E12, 124D12, 260H8, 268E9, 269A7,293H10, 370E5, 392D6, 402G3, 430H6, 485G10 and 487E3) were selected. Sequences of these clones are list in Table 1. Chimeric antibodies fused to human IgG1 Fc of these hybridoma were generated for further characterization.

TABLE-US-00004 TABLE 1 Antibody sequences selected from screening Anti- SEQ body ID chain Sequences* (CDR underlined and bold) NO: 4A6-VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDYYMH 1 WVKQRTEQGLEWIGR IDPDDGETKYAPKFQGKATMTADTSSNTAYLQLGS LTSEDAAVYYCARGG NYYVMDNWGQGTSVTVSS 4A6-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 2 WYQQKSGQSPKALIYT ASYRYSGVPDRFTGSGSGTDFTLTISNLQSEDLADYL CQQYFSYPHTFGG GTKLEIK 4B12-VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDSYMH 3 WVKERTEQGLEWIGR IDPEDGETNYAPKFQGKATLTADTSSNTAYLQLSG LTSEDTAVYYCARRV SSLYAMDYWGQGTSVTVSS 4B12-VL DIVMTQSQKSMSTSVGDRVSVTCKASQNVGTNVA 4 WYQQKPGQSPKPLIYL ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQYNNYPWTFGG GTKLEIK 4D8-VH EVRLQQSGAELVQPGASVKLSCTGFGFNIKDYYMH 5 WVKQRTEQGLEWIGR IDPEDAETKYAPKFQGQATITADTSSNTAYVQVSSL SSEDTAVYYCARGG NFYVMDYWGQGTSVTVSS 4D8-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVV 6 WYQQKPGQSPKVLIYS ASYRYSGVPDRFTGSGSGTDFTLIISNVQSEDLAEYF CQQKNSYPYTFGG GTKLEIK 8G11-VH EVQLQQSGAELVKPGASVKLSCTGFGFNIKDYYMH 7 WVKQRAEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQVSSL TSEDTAVYYCARGG NFYVMDYWGQGTSVTVSS 8G11-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVV 8 WYQQKPGQSPKVLIYS ASYRVSGVPDRFTGSGSGTDFTLIISNVQSEDLAEYF CQQKNNYPYTFGG GTKLEIK 12G11-VH AVQLQQSGAELVKPGASVKLSCTASGFNIKDYYMH 9 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLSSL TSEDTAVYYCARGG NYYAMDYWGQGTSVTVSS 12G11-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 10 WYQQKPGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQHNSYTYKFGG GTKVEIK 17F9-VH EVQLLQSWADLVKPGASVKLSCTASGFNIKDYYIH 11 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQDKAAITADTSSNTAYLQLSSL TSEGTAVYYCARYD GYYGFDYWGQGTTLTVSS 17F9-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 12 WYQQKPGQSPKPLIYS TSYRYSGVPDRFTGSGSGTDFTLTITNVQSEDLAEYF CHQYNNYPYTFGG GTKLEIK 18B6-VH EVQLQQSGAKLVKPGASVKLSCTASGFNIKDYYMC 13 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTASLQLSSL TSEDTAVYYCARGG NYYAMDYWGQGTSVTVSS 18B6-VL DTVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 14 WYQQKPGQSPKALIYS ATYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQHNSYSYTFGG GTKLEIK 18C1-VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDYYMH 15 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLSSL TSEDTAVYYCARLG NWVFDYWGQGTTLTVSS 18C1-VL DTVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVD 16 WYQQKSGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQYNTYTFGGGT KLEIK 19B6-VH EVQLQQFGAELVKPGASVKLSCTASGFNIKDYYMH 17 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQVKATITADTSSNTAYLHFSSL TSEDTAVYYCVRGG NFYYFDYWGQGTTLTVSS 19B6-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 18 WYQQKPGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQCINYPYTFGG GTKLEIK 20B3-VH EVHLQQSGAELVKPGTSLKLSCTASGFNIKDYYIHW 19 VKQRTEQGLEWIGR IDPEDGETKYAPTFQGKATITADTSSNTAYLQLSSL TSEDTAVYYCARYN GYSGFDYWGQGTTLTVSS 20B3-VL DIVMTQSQKFMSTSEGDRVSVTCKASQNVGTNVV 20 WYQQKPGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQYNRYPFTFGA GTKLELK 20E10-VH EVQLLQSWADLVKPGASVKLSCTASGFNIKDYYIH 21 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQDKAAITADTSSNTAYLQLSSL TSEGTAVYYCARYD GYYGFDYWGQGTTLTVSS 20E10-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 22 WYQQKPGQSPKPLIYS TSYRYSGVPDRFTGSGSGTDFTLTITNVQSEDLAEYF CHQYNNYPYTFGG GTKLEIK 20E12-VH EVQLQQSGAEVVKPGASVKLSCTASGFNIKDYYMH 23 WVKQRTEQGLECIGR IDPEDGETKYVPKFQGKATITAETSSNTAYLQLSSL TAEDTAVYYCSRGG SYYVMDYWGQGTSVTVSS 20E12-VL DVVMTQSRKFMSTSVGDRVSVTCKASQNVGTSVA 24 WYQQKLGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQDNSYPHTFGG GTKLEIK 21D6-VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDYYMH 25 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLSSL TSEDTAVYYCTSLH WSLDSWGQGTTLTVSS 21D6-VL DIVMTQSQKFMSTTVGDRVSITCKASQNVGTAVAW 26 YQQQPGQSPKPLIYS TANRDTGVPDRFTGSGSGTDFTLTISNMQSEDLAHY FCQQYSSYPYTFGG GTKLEIK 24F6-VH DVQLQQSGAELVKPGASVNLSCTGSGFNIKDYYIH 27 WVKQRTEQGLEWIGR VDPEDGETKYVPKFLDKATITADTSSNTAYLQLSSL TSEDTAVYYCTRGG NYYAMDYWGQGTSVTVSS 24F6-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 28 WYQQKSGQSPKALIFL ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQCNNYRLTFGS GTKLEIK 29E12-VH EVQLLQSWADLVKPGASVKLSCTASGFNIKDYYIH 29 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQDKAAITADTSSNTAYLQLSSL TSEGTAVYYCARYD GYYGFDYWGQGTTLTVSS 29E12-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 30 WYQQKPGQSPKPLIYS TSYRYSGVPDRFTGSGSGTDFTLTITNVQSEDLAEYF CHQYNNYPYTFGG GTKLEIK 30B5-VH EAQLQQSGAELVKPGASVKLSCTASGFNIRDSYIHW 31 VNQRTEQGLEWIGR IDPEDGETKYAPKFQGKATMTADTSSNTAYLQLSS LTSEDTAVYYCASWL ADYSAMDNWGQGTSVTVSS 30B5-VL DIQMTQSSSSFSVSLGDRVTITCKASEDIYNRLAWY 32 QQKPGNAPRLLISG ATSLETGVPSRFSGSGSGKDYTLSITSLQTEDVATYY CQQYWNTLYTFGG GTKLEMK 3008-VH EVQLQQSGAELVKPGASVKLSCTSSGFNIKDSYMH 33 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLNSL TSEDTAVYYCARRG SSLYAVDYWGQGTSVTVSS 3008-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTSVA 34 WYQQKPGQSPKAVIYL ASYRHRGVPARFTGSGSGTDFTLTISNVQSEDLAEY FCQQFNIYPWTFGG GTKLEIK 34H8-VH EVQLLQSGAELVKPGASVRLSCTASGFNIKDYYLH 35 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITTDTSSNTAYLQLSSL TSEDTAVYYCTRGG NYDVMDYWGQGTSVTVSS 34H8-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTYVV 36 WYQQKPGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTIGNVQSEDLAEYF CQQKNTYPFTFGG GTKLEIE 36E3-VH EVQLQQSGAELVKPGASVKLSCTSSGFNIKDSYMH 37 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLNSL TSEDTAVYYCARRG SSLYAVDYWGQGTSVTVSS 36E3-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTSVA 38 WYQQKPGQSPKAVIYL ASYRHRGVPARFTGSGSGTDFTLTISNVQSEDLAEY FCQQFNIYPWTFGG GTKLEIK 39F5-VH EVQLQQSGADLVRPGASVKLSCTASGFNIKDYYIHW 39 VKQRTEQGLEWIGR IDPEDGETKYAPKFQVKTTITADTSSNTAYLQFSSL TSEDTAVYYCVRGG NFYYFDYWGQGSTLTVSS 39F5-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 40 WYQQKPGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQCINYPYTFGG GTKLEIK 41F1-VH EVQLQQSGAELVKSGASVRLSCTASGFNIKDYYMH 41 WVKQRTEKGLEWIGR IDPEDGETKYVPKFQGKATITADTSSNTVYLQLNSL

TSEDTAVYYCVRGG NYYVMDYWGQGTSVTVSS 41F1-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTYVA 42 WYQQKPGQSPKVVIYS ASYRYNGVPDRFTGSGSGTDFTLTISNVQPEDLAEYF CQQYNNYPLTFGS GTKLEIK 46A10-VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDYYMH 43 WVKQRTEQGLELIGR IDPEDGETKYAPKFQVKATITADASSNTAYLQFSSL TSEDAAVYYCVRGG NFYYFDFWGQGTTLTVSS 46A10-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 44 WYQQKPGQSPKALVYS ASYRYSGVPDRFTGSGSGTDFTLTISDVQSEDLAEYF CQQCINYPYTFGG GTKLEIK 47C6-VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDYYIHW 45 VKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLHLSSL TSEDTAVYYCSRGG NFYYFDYWGQGTSLTVSS 47C6-VL DIVMTQSQKFMSTLVGDRVSVTCKASQNVGTNVA 46 WYQQKPGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQCNSYSYTFGG GTKLEIK 47F5-VH EVQLQQSGAEVVKPGASVKLSCTASGFNIKDYYMH 47 WVKQRTEQGLECIGR IDPEDGETKYVPKFQGKATITAETSSNTAYLQLSSL TAEDTAVYYCSRGG SYYVMDYWGQGTSVTVSS 47F5-VL DVVMTQSRKFMSTSVGDRVSVTCKASQNVGTSVA 48 WYQQKLGQSPKALIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQDNSYPHTFGG GTKLEIK 104A3-VH EVQLQQSGAELVKPGASVKVSCTGSGFNIKDYYIH 49 WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITSDTSSNTAYLQLSSL TSGDTAVYFCARYD GYYCFDYWGQGTTLTVSS 104A3-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 50 WYQQKPGQSPKPLIYS TSYRYSGVPDRFTGSGSGTDFTLTISNVQSADLAAYF CQQYNNYPYTFGG GTRLEIK 106A7-VH EVQLQQSGADLVKPGASVKLSCTTSGFNIKDYYMH 51 WVNQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLSSL TSEDTAVYYCARDW GHSFDYWGQGTTLTVSS 106A7-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTTVA 52 WYQQKPGQSPKTLIYS ASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQYNSYTFGGGT KLEMK 107E12- EVQLQQSGAEFVKPGASVKLSCTASGFNIKDYYIHW 53 VH VTQKTEQGLEWIGR IDPEDGETKYAPKFQDKATITADSSSNTAYLQLSSL TSVDTAVYFCSREG SFTGWFPYWGQGTLVSVSA 370E5-VL DIVMTQSQKFMSTSVGDRVSVTCKARQSVGTYVA 54 WYQQKPGQSPKALIYS TSYRYNGVPDRFTGSGSGTDFTLTISNVQSEDLADY FCQQYHSYPYTFGG GTKLEIK 124D12- EVQLQQSGAELVKPGASVKVSCTGSGFNIKDYYIH 55 VH WVKQRTEQGLEWIGR IDPEDGETKYAPKFQGRATITSDTSSNTAYLQLSSL TSGDTAVYYCARYD GYYCFDYWGQGTTLTVSS 124D12- DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 56 VL WYQQKPGQSPKPLIYS TSYRYSGVPDRFTGSGSGTDFTLTISNVQSADLAAYF CQQYNNYPYTFGG GTRLEIK 260H8-VH EVQLQESGPGLAKPSQTLSLTCSVTGYSITSDYWNW 57 IRKFPGNKLEYMGY ISYSGSIYYNPSLKSRISITRDTSKNQYYLQLNSVTNE DTATYYCARSGG MYYFDYWGQGTTLTVSS 260H8-VL DIQMTQSPASLSASVGETVTITCRASGNIHNYLAWY 58 QQKQGKSPQLLVYN AKTLEDGVPSRFSGSGSGTQYSLKINSLQPEDFGSYY CQHFWSIPPTFGS GTKLEIK 268E9-VH EVQLQESGPGLAKPSQTLSLTCSVTGYSITSDYWNW 59 IRKFPGNKLEYMGY ISYSGSIYYNPSLKSRISITRATSKNQYYLQLNSVTNE DTATYYCARSGG MYYFDYWGQGTTLTVSS 268E9-VL DIQMTQSPASLSASVGETVTITCRASGNIHNYLAWY 60 QQKQGKSPQLLVYN AKTLEDGVPSRFSGSGSGTQYSLKINSLQPEDFGSYY CQHFWSIPPTFGS GTKLEIK 269A7-VH EVQLQESGPGLAKPSQTLSLTCSVTGYSITSDYWNW 61 IRKFPGNKLEYMGY ISYSGTIYYNPSLKSRISITRDTSKNQYYLQLNSVTN EDTATYYCARSGG MYYFDYWGQGTTLTVSS 269A7-VL DIQMTQSPASLSASVGETVTITCRASGNIHNYLAWY 62 QQKQGKSPQLLVYN AKTLEDGVPSRFSGSGSGTQYSLKINSLQPEDFGSYY CQHFWSIPPTFGS GTKLEIK 293H10- EVQLQESGPGLAKPSQTLSLTCSVTGYSITSDYWNW 63 VH IRKFPGNKLEYMGY ISYSGNTDYNPSLKSRFSITRDTSKNQFYLQLNSVTT EDTATYYCARSEG MYFFDYWGQGTTLTVSS 293H10- DIQMTQSPASLSASVGETVSITCRASGNIHNYLAWY 64 VL HQKQGKSPQLLVYN AKTLADGVPSRFSGSGSGTQYSLKINSLQPEDFGSY YCQHFWSTPPTFGS GTKLEIK 370E5-VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDYYIHW 65 VKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLSSL TSEDTAVYYCARFG GLTAMDYWGQGTSVTVSS 370E5-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 66 WYQQKPGQSPKALIYA TSYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQYNNYPYTFGG GTKLEIK 392D6-VH AVQLQQSGTELVKPGASVKLSCSASGFNIKDYYVH 67 WVKQKTEQGLEWIGR IDPEDGETKYAPKFQGKATVTADTSSNTAYMQLSS LTSEDTAVYYCTRFG GLDAMDYWGQGTSVTVSS 392D6-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVA 68 WYQQKPGQSPKPLIYS TSYRYNGVPDRFTGSGSGTEFTLTISNVQSEDLAEYF CQQFNRYPYTFGG GTKLEIK 402G3-VH QVQLQQSGAELVKPGSSVKISCKASGFTFTSHFIHWI 69 KQQPGNGLKWIGW IYPGDDDTEYNHKFNGKATLTADKSSSTAYMHLSS LTSEDSAVYFCARRV EYYNGGFAYWGQGTLVTVSS 402G3-VL DVQMTQSPSYLAAPPGESVSISCKASKNIRNNLGWY 70 QERPGKTPNLLIHS GSTLQSGAPSRFSGGGSGTDFTLTIRSLESEDSAVYY CQQYDQYPLTFGS GTKLEIK 430H6-VH QVTLKESGPGILQPSQTLSLTCTFSGFSLNTYGMGV 71 GWIRQPSGKGLEWL ANIWWDDDKYYNPSLKNRLTISKDTSNNQAFLKIT NVDTADTATYFCARH PLPGYKDNYVVDAWGQGASVTVSS 430H6-VL DVVLTQTPGSLSVTLGDQASISCRSSCISLEYSDQYT 72 YLEWYLQKSGQSPQ LLIYGVSNRFSGVPDRFIGSGSGTDFTLKISRVEPEDL GVYYCFQATHDP YTFGAGTKLELK 485G10- EVQLQQSGAELVKPGASVKLSCTGSGFNIKDYYIHW 73 VH VKQRTEQGLEWIGR IDPEDGETKYAPKFQGKATITADTSSNTAYLQLSSL TSEDTVVYYCGRYH GYWALDYWGQGTSVTVSS 485G10- DIVMTQSQKFMSTSVGDRVSVTCKTSQNVGTNVA 74 VL WYQQKPGQSPKPLIYS TSYRYSGVPDRFTGSGSGTDFTLIISNVQSEDLAEYF CHQYFSYPYTFGG GTLLEIK 487E3-VH EVQLQQSGAELVKPGASVKLSCSASGFNIKDCYIHW 75 VKQRTEQGLEWIGR IDPEDGETKYAPKFQAKATITADTSSNTAYLQLNSL TSEDTAVYYCARHC NFLYFDYWGQGSTLTVS 487E3-VL DIVMTQSQKSMSTSLGDRVTVTCKASQNVGTIVAW 76 YQLKPGQSPKTLIYS ASYRSSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYF CQQYNNYPVIFGS GTKLEIR

Example 2. Binding Properties of Anti-IFNAR-1 Mouse Monoclonal Antibodies

[0120] This example tested the binding properties of the anti-IFNAR-1 mouse antibodies to the human IFNAR-1 proteins by ELISA assay. In short, His-tagged IFNAR-1 was coated at 0.5 .mu.g/ml overnight and then blocked by 5% BSA in PBS. Serial diluted anti-IFNAR-1 antibodies were incubated with the coated antigen for 1 h at room temperature. The resulting plates were washed with PBS/T and incubated with goat anti-mouse IgG-HRP for 1 h at room temperature. The plates were developed with TMB substrate and analyzed by spectrophotometer at OD 450-630 nm. The results of the ELISA assays are summarized in Table 2, which shows EC.sub.50 of binding to human IFNAR-1 protein.

TABLE-US-00005 TABLE 2 Binding EC.sub.50(nM) on hIFNAR-1 protein. Antibody EC.sub.50 (nM) Antibody EC.sub.50 (nM) 4D8 0.067 269A7 0.476 8G11 0.069 293H10 0.097 34H8 0.064 370 E5 0.052 104A3 0.056 392D6 0.054 106A7 0.099 402G3 0.137 107 E12 0.075 430H6 0.733 124D12 0.062 485G10 0.015 260H8 0.055 487 E3 0.028 268 E9 0.088

Anti-IFNAR-1 Antibody BIACORE Analysis

[0121] The binding of the antibodies to recombinant His-tagged human IFNAR-1-ECD protein was examined by Biacore T200 using a capture method. The anti-IFNAR-1 antibodies were captured using anti-human Fc antibody or Protein A which were coated on chip. The serial concentrations of his-tagged human IFNAR-1-ECD protein (0-8 nM) were injected over capture antibodies at the flow rate of 30 .mu.l/min. The dissociation phases were 600 s or 1200 s. The results are shown in Table 3 below. The Biacore results for the anti-IFNAR-1 antibodies demonstrate that these anti-IFNAR-1 antibodies are high affinity binders to human IFNAR-1.

TABLE-US-00006 TABLE 3 Binding of antibodies to recombinant IFNAR-1 protein Antibody ka (1/Ms) kd (1/s) KD (M) 4D8 2.31E+06 9.03E-05 3.90E-11 8G11 2.36E+06 6.40E-05 2.71E-11 34H8 2.37E+06 4.41E-05 1.86E-11

Example 3. IFN-Responsive Reporter Assay for Screening Anti-IFNAR-1 Mouse Monoclonal Antibodies

[0122] HEK-Blue.TM. IFN.alpha./.beta. cells (InvivoGen) are specifically designed to monitor the activation of the JAK-STAT pathway induced by type I IFNs. Upon IFN-.alpha. or IFN-.beta. stimulation, these cells activate the JAK-STAT pathway and subsequently the expression of the reporter gene named secreted embryonic alkaline phosphatase (SEAP). SEAP which is secreted in the supernatant is easily detectable by using a SEAP detection reagent QUANTI-Blue.TM. (InvivoGen). A neutralizing antibody, which blocks interferon binding to its receptor, will inhibit IFN-induced reporter gene expression. This reporter assay was utilized for screening anti-IFNAR-1 mouse monoclonal antibodies.

[0123] HEK-Blue.TM. IFN.alpha./.beta. cells were incubated with 400 U/ml IFN.alpha.2b in the presence of serial dilutions of anti-IFNAR-1 mouse monoclonal antibodies overnight. The expression of reporter gene SEAP was determined by using a spectrophotometer at 650 nm. EC.sub.50 of the tested antibodies are listed in Table 4. Among these antibodies, 4D8, 8G11, 34H8, 485G10 and 487E3 antibodies showed superior effects in blocking IFN.alpha.2b signal. Given the analysis on sequence homology, the clones of 8G11 and 485G10 were selected for further humanization and characterization.

TABLE-US-00007 TABLE 4 Reporter assay for the antibodies Clone ID EC.sub.50 (nM) Clone ID EC.sub.50(nM) 4D8 0.460 286E9 3.871 8G11 0.465 293H10 1.285 34H8 0.560 370E5 2.170 104A3 1.310 392D6 0.824 106A7 3.516 402G3 1.267 107E12 1.207 430H6 1.297 124D12 0.981 485G10 0.286 260H8 5.074 487E3 0.161 269A7 6.713

Example 4. Anti-IFNAR-1 mAb Humanization

A. 8G11

[0124] The mouse antibody 8G11 variable region genes were employed to create a humanized MAb. In the first step of this process, the amino acid sequences of the VH and VK of 8G11 were compared against the available database of human Ig gene sequences to identify the overall best-matching human germline Ig gene sequences. For the heavy chain, the closest human match was the IGHV1-46*01 gene. For the light chain, the best human match was the IGKV1-9*01 gene.

[0125] Humanized variable domain sequences were then designed where the CDR1 (SEQ ID NO:77), 2 (SEQ ID NO:78), and 3 (SEQ ID NO:80) sequences of the 8G11 VH were grafted onto framework sequences of the IGHV1-46*01 gene and the CDR1 (SEQ ID NO:81), 2 (SEQ ID NO:82) and 3 (SEQ ID NO:83) of the 8G11 light chain were grafted onto framework sequences of the IGKV1-9*01 gene. A 3D model was then generated to determine if there were any framework positions where replacing the mouse amino acid to the human amino acid could affect binding and/or CDR conformation. In the case of the heavy chain, K12, V20, A24, R38, M48, V68, M70, R72, V79 and M81 (Kabat numbering) in human framework was identified and subjected to back-mutations to their moue counterpart amino acid i.e.: K12V, V20L, A24G, R38K, M48I, V68A, M70I, R72A, V79A and M81L. In the case of the light chain, L4, A13, 121, A43, L46, T74, L78 and Y87 (Kabat numbering) in human framework was identified and subjected to back-mutation to their moue counterpart amino acid i.e.: L4M, A13T, I21V, A43S, L46V, T74I, L78V and Y87F. At the meantime, G56 A mutation was employed to remove the PTM.

TABLE-US-00008 TABLE 5 8G11 sequences and CDRs Antibody chain SEQ ID or domain Sequences (CDR underlined and bold) NO: 8G11-VH EVQLQQSGAELVKPGASVKLSCTGFGFNIKDYYMHWVKQRAEQGLEWIGR 7 IDPEDGETKYAPKFQGKATITADTSSNTAYLQVSSLTSEDTAVYYCARGG NFYVMDYWGQGTSVTVSS 8G11-VL DIVMTQSQKFMSTSVGDRVSVTCKASQNVGTNVVWYQQKPGQSPKVLIYS 8 ASYRVSGVPDRFTGSGSGTDFTLIISNVQSEDLAEYFCQQKNNYPYTFGG GTKLEIK CDRH1 DYYMH 77 CDRH2 RIDPEDGETKYAPKFQG 78 CDRH2 RIDPEDAETKYAPKFQG 79 (modified) CDRH3 GGNFYVMDY 80 CDRL1 KASQNVGTNVV 81 CDRL2 SASYRVS 82 CDRL3 QQKNNYPYT 83

[0126] The humanized sequences are listed in Table 6: 8G11-VH1, 8G11-VH2, 8G11-VH3, 8G11-VH4, 8G11-VL1, 8G11-VL2, 8G11-VL3, and 8G11-VL4.

TABLE-US-00009 TABLE 6 Humanized sequences Antibody SEQ ID chain Sequences (CDR italic; back mutations bold and underlined) NO: 8G11-VH1 QVQLVQSGAEVKKPGASVKVSCKASGFNIKDYYMHWVRQAPGQGLEWMGR 84 IDPED ETKYAPKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGG NFYVMDYWGQGTLVTVSS 8G11-VH2 QVQLVQSGAEVVKPGASVKVSCKGSGFNIKDYYMHWVRQAPGQGLEWIGR 85 IDPED ETKYAPKFQGRVTMTADTSTSTVYMELSSLRSEDTAVYYCARGG NFYVMDYWGQGTLVTVSS 8G11-VH3 QVQLVQSGAEVVKPGASVKVSCKGSGFNIKDYYMHWVKQAPGQGLEWIGR 86 IDPED ETKYAPKFQGRVTITADTSTSTVYLELSSLRSEDTAVYYCARGG NFYVMDYWGQGTLVTVSS 8G11-VH4 QVQLVQSGAEVVKPGASVKLSCKGSGFNIKDYYMHWVKQAPGQGLEWIGR 87 IDPED ETKYAPKFQGRATITADTSTSTAYLELSSLRSEDTAVYYCARGG NFYVMDYWGQGTLVTVSS 8G11-VL1 DIQLTQSPSFLSASVGDRVTITCKASQNVGTNVVWYQQKPGKAPKLLIYS 88 ASYRVSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQKNNYPYTFGQ GTKLEIK 8G11-VL2 DIQLTQSPSFLSTSVGDRVTITCKASQNVGTNVVWYQQKPGKSPKVLIYS 89 ASYRVSGVPSRFSGSGSGTEFTLIISSLQPEDFATYYCQQKNNYPYTFGQ GTKLEIK 8G11-VL3 DIQLTQSPSFLSTSVGDRVTITCKASQNVGTNVVWYQQKPGKSPKVLIYS 90 ASYRVSGVPSRFSGSGSGTEFTLIISSVQPEDFATYFCQQKNNYPYTFGQ GTKLEIK 8G11-VL4 DIQMTQSPSFLSTSVGDRVTVTCKASQNVGTNVVWYQQKPGKSPKVLIYS 91 ASYRVSGVPSRFSGSGSGTEFTLIISSVQPEDFATYFCQQKNNYPYTFGQ GTKLEIK

[0127] All the 16 IgGs were expressed in the HEK293 cell line. Express antibodies were performed for affinity ranking. Detailed data of affinity ranking were summarized in Table 7.

TABLE-US-00010 TABLE 7 Affinity ranking of humanized antibodies. Antibody k.sub.a (1/Ms) k.sub.d (1/s) K.sub.D (M) VH4 + VL3 3.01E+05 8.52E-05 2.84E-10 VH2 + VL2 2.87E+05 9.42E-05 3.28E-10 VH4 + VL2 2.92E+05 9.69E-05 3.32E-10 VH3 + VL3 3.01E+05 1.10E-04 3.64E-10 VH2 + VL3 2.65E+05 9.65E-05 3.64E-10 VH4 + VL4 2.83E+05 1.05E-04 3.72E-10 VH2 + VL4 2.62E+05 1.02E-04 3.88E-10 VH3 + VL2 2.84E+05 1.11E-04 3.89E-10 VH3 + VL4 2.81E+05 1.13E-04 4.01E-10 VH3 + VL1 2.56E+05 1.06E-04 4.13E-10 VH2 + VL1 2.02E+05 8.84E-05 4.39E-10 VH4 + VL1 2.03E+05 9.94E-05 4.89E-10 VH1 + VL1 1.48E+05 9.83E-05 6.62E-10 VH1 + VL2 1.67E+05 1.27E-04 7.65E-10 VH1 + VL4 1.69E+05 1.41E-04 8.36E-10 VH1 + VL3 2.31E+05 2.00E-04 8.65E-10

[0128] Based on the affinity ranking results, 4 IgGs (VH4+VL3, VH2+VL2, VH2+VL1, VH1+VL1) were expressed and purified. The purified antibodies were further selected for affinity measurement under different concentrations. Detailed data were summarized in Table 8.

TABLE-US-00011 TABLE 8 Affinity of selected humanized antibodies. Antibody ka (1/Ms) kd (1/s) KD (M) 8G11-H4L3 3.40E+05 7.47E-05 2.20E-10 8G11-H2L2 2.98E+05 1.25E-04 4.19E-10 8G11-H2L1 2.29E+05 9.24E-05 4.03E-10 8G11-H1L1 1.61E+05 2.19E-04 1.36E-09

B. 485G10

[0129] The mouse antibody 485G10 variable region genes were employed to create a humanized MAb. In the first step of this process, the amino acid sequences of the VH and VK of 485G10 were compared against the available database of human Ig gene sequences to identify the overall best-matching human germline Ig gene sequences. For the heavy chain, the closest human match was the IGHV1-2*05 gene. For the light chain, the best human match was the IGKV1-16*01 gene.

[0130] Humanized variable domain sequences were then designed where the CDR1 (SEQ ID NO:92), 2 (SEQ ID NO:93), and 3 (SEQ ID NO:95) sequences of the 485G10 heavy chain were grafted onto framework sequences of the IGHV1-2*05 gene and the CDR1 (SEQ ID NO:96), 2 (SEQ ID NO:C5) and 3 (SEQ ID NO:98) of the 485G10 light chain were grafted onto framework sequences of the IGKV1-16*01 gene. A 3D model was then generated to determine if there were any framework positions where replacing the mouse amino acid to the human amino acid could affect binding and/or CDR conformation. In the case of the heavy chain, V20, A24, R38, M48, V68, M70, R72, M81, A97 (Kabat numbering) in human framework was identified and subjected to back-mutations to their moue counterpart amino acid i.e.: V20L, A24G, R38K, M48I, V68A, M70I, R72A, M81L, A97G. In the case of the light chain, A13, I21, F36, S46, L78, V104 (Kabat numbering) in human framework was identified and subjected to back-mutation to their moue counterpart amino acid i.e.: A13T, I21V, F36Y, S46P, L78V, V104L. At the meantime, G56 A mutation was employed to remove the PTM.

TABLE-US-00012 TABLE 9 485G10 sequences and CDRs Anti- body chain SEQ or ID domain Sequences (CDR underlined and bold) NO: 485G10- EVQLQQSGAELVKPGASVKLSCTGSGFNIK 73 VH DYYIHWVKQRTEQGLEWIGRIDPEDGETKY APKFQGKATITADTSSNTAYLQLSSLTSED TVVYYCGRYHGYWALDYWGQGTSVTVSS 485G10- DIVMTQSQKFMSTSVGDRVSVTCKTSQNV 74 VL GTNVAWYQQKPGQSPKPLIYSTSYRYSGVP DRFTGSGSGTDFTLIISNVQSEDLAEYFC HQYFSYPYTFGGGTLLEIK CDRH1 DYYIH 92 CDRH2 RIDPEDGETKYAPKFQG 93 CDRH2 RIDPEDAETKYAPKFQG 94 (modi- fied) CDRH3 YHGYWALDY 95 CDRL1 KTSQNVGTNVA 96 CDRL2 STSYRYS 97 CDRL3 HQYFSYPYT 98

[0131] The humanized sequences are listed in Table 10: 485G10-VH0, 485G10-VH1, 485G10-VH2, 485G10-VH3, 485G10-VL1, 485G10-VL2, 485G10-VL3, and 485G10-VL4.

TABLE-US-00013 TABLE 10 Humanized sequences Anti- SEQ body Sequences (CDR italic and underlined; ID chain back mutations bold) NO: 485G10- QVQLVQSGAEVKKPGASVKVSCKASGFNIKDYYIHWVRQAPGQGLEWMGR 99 VH0 IDPED ETKYAPKFQGRVTMTRDTSISTAYMELSRLRSDDTVVYYCARYH GYWALDYWGQGTLVTVSS 485G10- QVQLVQSGAEVKKPGASVKVSCKASGFNIKDYYIHWVRQAPGQGLEWMGR 100 VH1 IDPED ETKYAPKFQGRVTMTADTSISTAYMELSRLRSDDTVVYYCARYH GYWALDYWGQGTLVTVSS 485G10- QVQLVQSGAEVKKPGASVKLSCKASGFNIKDYYIHWVKQAPGQGLEWIGR 101 VH2 IDPED ETKYAPKFQGRVTMTADTSISTAYMELSRLRSDDTVVYYCGRYH GYWALDYWGQGTLVTVSS 485G10- QVQLVQSGAEVKKPGASVKLSCKGSGFNIKDYYIHWVKQAPGQGLEWIGR 102 VH3 IDPED ETKYAPKFQGRATITADTSISTAYLELSRLRSDDTVVYYCGRYH GYWALDYWGQGTLVTVSS 485G10- DIQMTQSPSSLSASVGDRVTITCKTSQNVGTNVAWFQQKPGKAPKSLIYS 103 VL1 TSYRYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYFSYPYTFGP GTKVDIK 485G10- DIQMTQSPSSLSTSVGDRVTITCKTSQNVGTNVAWYQQKPGKAPKPLIYS 104 VL2 TSYRYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYFSYPYTFGP GTKVDIK 485G10- DIQMTQSPSSLSTSVGDRVTITCKTSQNVGTNVAWYQQKPGKAPKPLIYS 105 VL3 TSYRYSGVPSRFSGSGSGTDFTLTISSVQPEDFATYYCHQYFSYPYTFGP GTKLDIK 485G10- DIQMTQSPSSLSTSVGDRVTVTCKTSQNVGTNVAWYQQKPGKAPKPLIYS 106 VL4 TSYRYSGVPSRFSGSGSGTDFTLTISSVQPEDFATYYCHQYFSYPYTFGP GTKLDIK

[0132] All the 16 IgGs could be expressed in the HEK293 cell line. Express antibodies were performed for affinity ranking. Detailed data of affinity ranking were summarized in table 11.

TABLE-US-00014 TABLE 11 Affinity ranking of humanized antibodies. Antibody k.sub.a (1/Ms) k.sub.d (1/s) K.sub.D (M) VH1 + VL3 3.11E+05 1.01E-04 3.25E-10 VH1 + VL2 3.08E+05 2.01E-04 6.52E-10 VH1 + VL4 3.27E+05 2.16E-04 6.60E-10 VH2 + VL3 2.81E+05 2.38E-04 8.48E-10 VH3 + VL2 3.06E+05 2.64E-04 8.64E-10 VH3 + VL4 3.54E+05 2.71E-04 7.66E-10 VH3 + VL3 3.32E+05 2.79E-04 8.41E-10 VH2 + VL2 3.29E+05 2.98E-04 9.06E-10 VH2 + VL4 3.03E+05 3.00E-04 9.90E-10 VH0 + VL3 2.94E+05 3.16E-04 1.07E-09 VH0 + VL4 2.55E+05 3.18E-04 1.25E-09 VH0 + VL2 2.79E+05 4.39E-04 1.57E-09 VH1 + VL1 3.28E+05 1.03E-03 3.14E-09 VH0 + VL1 5.09E+05 4.75E-03 9.33E-09 VH2 + VL1 3.23E+05 6.63E-03 2.05E-08 VH3 + VL1 9.71E+05 1.85E-02 1.91E-08

[0133] Based on the affinity ranking results, 4 IgGs (VH1+VL2, VH1+VL4, VH1+VL3, VH2+VL3) were expressed and purified. The purified antibodies were further selected for affinity measurement under different concentrations. Detailed data were summarized in Table 12.

TABLE-US-00015 TABLE 12 Affinity of selected humanized antibodies. Antibody ka (1/Ms) kd (1/s) KD (M) 485G10-H1L2 2.07E+05 3.21E-04 1.55E-09 485G10-H1L4 1.96E+05 2.59E-04 1.32E-09 485G10-H1L3 1.87E+05 2.32E-04 1.24E-09 485G10-H2L3 1.86E+05 3.66E-04 1.96E-09

Example 5. Anit-IFNAR-1 Humanized Monoclonal Antibodies Inhibit the Biological Activity of Interferon .alpha.2b in IFN-Responsive Reporter Assay

[0134] To evaluate the IFNAR-1-blocking function of humanized antibodies, in vitro IFN-responsive reporter assay described in Example 3 was used. HEK-Blue.TM. IFN.alpha./.beta. cells were incubated with 400 U/ml IFN.alpha.2b in the presence of serial diluted anti-IFNAR-1 humanized monoclonal antibodies overnight prior to determination by using a spectrophotometer at 650 nM. As shown in FIG. 1, the 8G11H, and 485G10H antibodies efficiently blocked IFN.alpha.c2b-induced reporter gene expression. Their potency is comparable to that of the corresponding chimeric antibodies.

Example 6. Anit-IFNAR-1 Humanized Monoclonal Antibodies Inhibit the Biological Activity of Interferon .alpha.2b in Daudi Cell Proliferation

[0135] The cell line Daudi, derived from a human B-lymphoblast Burkitt's lymphoma, expresses high levels of IFNAR, and the growth of these cells is inhibited by type I interferons. To measure the functional blocking ability of humanized anti-IFNAR-1 antibodies, Daudi cells were cultured with interferon oc2b in the presence or absence of a serial diluted anti-IFNAR-1 humanized antibodies. The cell proliferation was measured by Cell Counting Kit-8 (CCK8, DOJINDO Laboratories). As described in FIG. 2, the 8G11H and 485G10H antibodies dose-dependently reverse IFN.alpha.c2b-mediated inhibition of Daudi cell proliferation. Their potency is comparable to that of the corresponding chimeric antibodies.

Example 7. Anti-IFNAR-1 Human Monoclonal Antibodies Inhibit the Biological Activity of Multiple Type I IFNs

[0136] To assess the ability of humanized anti-IFNAR-1 antibodies to inhibit the biological activity of multiple type I IFNs, distinct IFN alpha subtypes were tested in the IFN-responsive reporter assay. HEK-Blue.TM. IFN-.alpha./.beta. cells were incubated with one of the following IFN alpha subtypes: 1, 2a, 4, 5, 6, 7, 8, 10, 14, 16, 17 and 21 in the presence of serial dilutions of the tested antibodies or an isotype control. The reporter signal was determined as described in Example 3. As shown in Table 13, the 8G11H and 485G10H antibodies can efficiently inhibit the cell signal elicited by multiple type I IFNs. Their EC.sub.50 is in the pM level.

TABLE-US-00016 TABLE 13 Reporter assay of multiple Type I IFNs EC.sub.50 (nM) Clone ID 8G11H 485G10H IFN.alpha. 1 0.057 0.051 IFN.alpha. 2a 0.052 0.048 IFN.alpha. 4 0.055 0.054 IFN.alpha. 5 0.087 0.098 IFN.alpha. 6 0.090 0.072 IFN.alpha. 7 0.039 0.047 IFN.alpha. 8 0.604 0.490 IFN.alpha. 10 0.402 0.550 IFN.alpha. 14 0.215 0.153 IFN.alpha. 16 0.061 0.079 IFN.alpha. 17 0.035 0.035 IFN.alpha. 21 0.079 0.065

Example 8. Anti-IFNAR-1 Human Monoclonal Antibodies Inhibit Type I IFN Mediated Anti-Viral Function

[0137] Type I IFNs were firstly identified by their unique anti-viral function. To evaluate the blocking effects of the anti-IFNAR-1 antibodies on type I IFN-mediated anti-viral function, an IFN.alpha.2b-based anti-viral assay was established. Briefly, cell line Huh7 was engineered to contain the full-length HCV genotype 1b replicon (Con1b) which was linked to a firefly luciferase reporter gene. The abundance of reporter gene closely correlated with replication levels of HCV replicon. The activity of anti-IFNAR-1 antibodies in blocking IFN.alpha.2b induced anti-viral function can be determined by the restoration of reporter gene levels in IFN.alpha.c2b-treated Huh7-Con1b cells.

[0138] Huh7-Con1b cells were cultured with 400 U/ml IFN.alpha.2b in the presence or absence of serial diluted anti-IFNAR-1 humanized monoclonal antibodies for 48 hours. The expression of firefly luciferase reporter gene was measured by addition of Britelite plus reagent and further determined by using a spectrophotometer. As shown in Table 14, anti-IFNAR-1 humanized monoclonal antibodies 8G11H and 485G10H efficiently inhibit IFN.alpha.c2b-induced anti-viral function, their EC50 is comparable to that of the corresponding chimeric antibodies.

TABLE-US-00017 TABLE 14 IFN.alpha.2b-mediated anti-viral assay Clone ID EC.sub.50 (nM) 8G11-chimeric 7.733 8G11H 8.067 485G10-chimeric 16.267 485G10H 13.867

Example 9. Inhibition of Type I IFN-Induced IP-10 Secretion by Anti-IFNAR-1 Humanized Monoclonal Antibodies

[0139] Upon type I IFNs stimulation, normal peripheral blood mononuclear cells (PBMCs) express a serial of IFN-response genes including IP-10. The activity of anti-IFNAR-1 antibodies was tested for inhibition of IFN.alpha.c2b-induced secretion of IP-10 by normal PBMCs.

[0140] PBMCs were incubated with 400 U/ml IFN.alpha.2b in the presence or absence of anti-IFNAR-1 humanized monoclonal antibodies for 48 hours. Supernatants were collected and analyzed for IP-10 concentration by using a quantitative LANCE kit (Cisbio) according to manufacturer recommendations. As shown in FIG. 3, anti-IFNAR-1 humanized monoclonal antibodies 8G11H and 485G10H dose-dependently inhibit recombinant IFN.alpha.c2b-induced secretion of IP-10 by normal PBMC culture.

Example 10. Inhibition of Type I IFN-Induced Dendritic Cell Development by Anti-IFNAR-1 Humanized Monoclonal Antibodies

[0141] It is well known that type I IFNs induce the development and maturation of dendritic cells, as defined by the expression of specific cell surface markers such as CD38, CD80 and CD86 et al, and the capability to stimulate naive allogeneic CD4.sup.+ T cell proliferation (mixed lymphocyte reaction, MLR). To evaluate the function of anti-IFNAR-1 antibodies in blocking type I IFN signaling on dendritic cells differentiation, IFN.alpha.2b-mediated dendritic cells differentiation assay was established. Monocytes isolated from human PBMCs were cultured with 20 ng/ml GMCSF and 400 U/ml IFN.alpha.2b in the presence or absence of anti-IFNAR-1 humanized monoclonal antibodies for 72 hours. Monocyte-derived dendritic cells were collected and further cultured with human naive allogeneic CD4.sup.+ T cells at a ratio of 1:5 for 5 days. The cytokine IFN-.gamma., reflecting the proliferation status of CD4.sup.+ T cells, in the culture supernatants were analyzed by ELISA assay.

[0142] As shown in FIG. 4, the addition of anti-IFNAR-1 humanized monoclonal antibodies, 8G11H or 485G10H, into the differentiation systems results in significant reduction of cell surface markers CD38 and CD86 expression, as well as obvious impaired responses in MLR as demonstrated by decreased production of IFN-.gamma. by CD4+ T cells (FIG. 5). The results demonstrated that anti-IFNAR-1 humanized monoclonal antibodies, 8G11H and 485G10H, can efficiently block IFN.alpha.2b signaling on dendritic cells development.

Example 11. Inhibition of SLE Plasma Mediated Dendritic Cell Development by Anti-IFNAR-1 Humanized Monoclonal Antibodies

[0143] The capacity of SLE patients' plasma to induce dendritic cell development correlates with disease activity and depends on the actions of IFN.alpha.. In this example, the purified human anti-IFNAR-1 antibodies, 8G11H and 485G10H, were tested for inhibition of SLE plasma-induced dendritic cell development, as assessed by the impaired capability of antibody-treated dendritic cells to stimulate naive allogeneic CD4.sup.+ T cell proliferation (MLR).

[0144] Monocytes isolated from human PBMCs were cultured in media containing 25% SLE patient plasma in the presence or absence of anti-IFNAR-1 antibodies (15 .mu.g/ml) for three to five days. The differentiated cells were then collected and co-cultured with human naive allogeneic CD4.sup.+ T cells at a ratio of 1:5 for five days. The cytokine IFN-.gamma., reflecting the proliferation status of CD4.sup.+ T cells, in the culture supernatants were analyzed by ELISA assay. As shown in FIG. 6, anti-IFNAR-1 humanized monoclonal antibodies 8G11H and 485G10H significantly inhibit the IFN.alpha.-dependent dendritic cell development, as demonstrated by impaired capability of differentiated cells to stimulate MLR with marked reduction of the cytokine IFN-.gamma. production by CD4.sup.+ T cells.

Example 12. Bispecific Antibody to Target BAFF and IFNAR-1

[0145] This example tested whether certain formats of anti-BAFF/anti-IFNAR1 bispecific antibodies can retain the binding affinities, and inhibitive activities, to both antigens.

[0146] The bispecific monoclonal antibody was generated with Belimumab (anti-BAFF) and 8G11 chimeric antibody. FIG. 7 depicts a schematic of designed four formats for the anti-IFNAR/BAFF bispecific antibody (Bi-BFINR). The binding properties of Bi-BFINR to human IFNAR-1 protein and human BAFF protein were detected respectively by ELISA assay. As shown in FIG. 8, the format 1 of Bi-BFINF exhibited similar activity as 8G11 antibody in binding to IFNAR-1 and comparable activity to Belimumab in binding to BAFF. Furthermore, cell-based assay was performed to characterize the biological function of Bi-BFINF. For anti-IFNAR-1 arm, IFN.alpha.2b-based reporter assay was applied. As shown in FIG. 9A, Bi-BFINF antibody with format 1 has the comparable activity to chimeric 8G11 in blocking IFN.alpha.2b signaling. For anti-BAFF arm, BAFF-induced B cell proliferation assay was performed. B cells isolated from Tonsil were cultured with 2 .mu.g/ml recombinant BAFF in the presence or absence of Bi-BFINF for 72 hours. Cell-title glo was added to the cultures to determine B cell numbers. As shown in FIG. 9B, Bi-BFINF antibody with format 1 exhibits better activity in blockade of BAFF-induced B cell proliferation.

Example 13. Identification of Critical Amino Acids of hIFNAR-1 for 8G11 Binding

[0147] For epitope-mapping for the 8G11 antibody, a mutation library (alanine scan) of hIFNAR-1(1aa-409aa) was created. Binding of 8G11 Fab to each mutant clone in the alanine scanning library was determined, in duplicate, by high-throughput flow cytometry. For each point, background fluorescence was subtracted from the raw data, which were then normalized to Fab reactivity with WT target protein. For each mutant clone, the mean binding value was plotted as a function of expression (represented by control reactivity). 85288 Mab (R&D) and Anifrolumab were used as control antibody. To identify preliminary primary critical clones a threshold of >70% WT binding to control MAb or 8G11 Fab and <30% WT binding to test 8G11 Fab was applied.

[0148] Screening Results of the mutations are listed in Table 15. The clone H273A that did not meet the set thresholds but whose decreased binding activity and proximity to critical residues suggested that the mutated residue may be part of the antibody epitope. Critical residues whose mutation gave the lowest reactivities with specific antibodies are highlighted in bold and underlined. Validated critical residues represent amino acids whose side chains make the highest energetic contributions to the antibody-epitope interaction. Therefore, the highlighted (bold and underlined) residues K276, K278 are likely the major energetic contributors to binding. Results were showed in Table 16.

TABLE-US-00018 TABLE 15 Identification of critical residues for 8G11 Fab binding. Binding Reactivity (% WT) 8G11 85288 Mutation Fab Mab Anifrolumab Mab H273A 31.2 95.5 88.1 L274A 22.7 115.1 105.6 Y275A 19.1 86.8 145.5 K276A 1.5 104.4 99.4 K278A 3.9 101.2 105.1

TABLE-US-00019 TABLE 16 Important residues for binding of Fab to the hIFNAR-1 protein. Antibody Name Residues 8G11 H273, L274, Y275, K276, K278

[0149] Competition assays were used to test whether the antibodies 4D8, 8G11, 4B12, 30C8, 30B5, 34H8 and Anifrolumab competed with one another. The results showed that 4D8, 8G11, 4B12, 30C8, 30B5, and 34H8 competed with one another but none competed with Anifrolumab for binding to IFNAR-1. These data show that the newly identified antibodies target the same epitope on IFNAR-1 which is different from that for Anifrolumab.

Example 14. Pharmacokinetics of Humanized Anti-IFNAR1 Antibodies in Cynomolgus Monkeys

[0150] A preliminary pharmacokinetics (PK) study was conducted in cynomolgus monkeys at a dose of 10 mg/kg of 8G11-H2L1 and 485G10-H1L3. Serum samples were collected relative to antibody injection at -0 (predose), 5 Min, 30 Min, 1 h, 2h, 4h, 8h, 24 hr (1 day), 48 h (2 days), 72 hr (3 days), 96 hr (4 days), 120 hr (5 days), 168 hr (7 days), 240 hr (10 days), 36 hr (14 days), 504 hr (21 days), 672 hr (28 days) and 840 hr (35 days). The levels of anti-IFNAR1 mAbs were measured by a generic ELISA method using recombinant IFNAR1 protein ECD as the capture antigen. PK parameters were calculated based on statistical moment theory by WinNonlin 6.3 software. Results are shown in Table 17.

[0151] Time-concentration profiles indicated that the PK of 8G11-H2L1 and 485G10-H1L3 were more in line with what one would expect for a typical IgG (FIGS. 10A and 10B),

TABLE-US-00020 TABLE 17 8G11-H2L1 and 485G10-H1L3 PK parameters in cynomolgus monkeys i.v. 10 mg/kg 485G10-H1L3 8G11-H2L1 HL_Lambda_z h 41.9 71.37 T.sub.max h 0.08 0.08 C.sub.max .mu.g/mL 299.05 241.32 AUC.sub.last h * .mu.g/mL 12942.47 15450.93 Vz_F_obs mL/kg 46.4 65.6 Cl_F_obs mL/h/kg 0.77 0.64 MRT.sub.last h 62.43 99.49

Example 15. Pharmacodynamics of Humanized Anti-IFNAR1 Antibodies in Cynomolgus Monkeys

[0152] A pharmacodynamic (PD) model was to used study the ability of the anti-IFNAR1 antibodies to inhibit interferon activity in vivo. Cynomolgus monkeys were treated with i.v. infusion of 10 mg/kg 8G11-H2L1 and 485G10-H1L3 or vehicle control followed by i.m. dose of human IFN-.alpha.2b (3*10E6U/Kg). After treatment and IFN-.alpha.2b injection, blood was collected at pre-dose, 24, 72 and 168 h after treatment for PD marker measurement. The expression of CD11C, CD38, CD86, MHC Class I, MHC Class II and IFN-alpha R1 was detected by FACS in total PBMC. As shown in FIG. 11A, 8G11-H2L1 and 485G10-H1L3 could significantly decrease the HLA-DR expression of CD14+ cell at 24 h after treatment. Meanwhile, 8G11-H2L1 and 485G10-H1L3 were also able to suppress the IFN-alpha R1 expression of CD14+ cell both at 24 h and 72 h after treatment.

[0153] The serum levels of monkey neopterin and beta-2-microglobulin were detected by ELISA performed according to the manufacturer's instruction. The level of CRP in peripheral blood was measured by blood biochemistry. As shown in FIG. 11B, Neopterin, Beta-2 Microglobulin and CRP in vehicle group were increased rapidly within 24 h. Compared with vehicle control, 8G11-H2L1 and 485G10-H1L3 could suppress the neopterin and beta-2 Microglobulin significantly at 24 h and 72 h after dosing. Also, 8G11-H2L1 and 485G10-H1L3 were able to decrease the CRP significantly at 24 h after dosing.

[0154] The expression levels of type I IFN-induced gene signatures in peripheral blood mononuclear cells were detected by real-time PCR. As shown in FIG. 11C, the mRNA levels of ISG15, HERC5, IFI27 and IFIT3 were increased rapidly within 24 h in vehicle group. Compared with vehicle control, 8G11-H2L1 and 485G10-H1L3 significantly blocked the induction effects of type I IFN on these genes.

[0155] The present disclosure is not to be limited in scope by the specific embodiments described which are intended as single illustrations of individual aspects of the disclosure, and any compositions or methods which are functionally equivalent are within the scope of this disclosure. It will be apparent to those skilled in the art that various modifications and variations can be made in the methods and compositions of the present disclosure without departing from the spirit or scope of the disclosure. Thus, it is intended that the present disclosure cover the modifications and variations of this disclosure provided they come within the scope of the appended claims and their equivalents.

[0156] All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference

Sequence CWU 1

1

2061118PRTArtificial SequenceSynthetic 1Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Asp Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Met Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Gly Ser Leu Thr Ser Glu Asp Ala Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Tyr Tyr Val Met Asp Asn Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 1152107PRTArtificial SequenceSynthetic 2Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Thr Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Leu Gln Ser65 70 75 80Glu Asp Leu Ala Asp Tyr Leu Cys Gln Gln Tyr Phe Ser Tyr Pro His 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 1053119PRTArtificial SequenceSynthetic 3Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Ser 20 25 30Tyr Met His Trp Val Lys Glu Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Asn Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Leu Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Gly Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg Val Ser Ser Leu Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110Thr Ser Val Thr Val Ser Ser 1154107PRTArtificial SequenceSynthetic 4Asp Ile Val Met Thr Gln Ser Gln Lys Ser Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Leu Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Asn Tyr Pro Trp 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 1055118PRTArtificial SequenceSynthetic 5Glu Val Arg Leu Gln Gln Ser Gly Ala Glu Leu Val Gln Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Gly Phe Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Gln Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Val Gln Val Ser Ser Leu Ser Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Phe Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 1156107PRTArtificial SequenceSynthetic 6Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Val Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Ile Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Lys Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 1057118PRTArtificial SequenceSynthetic 7Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Gly Phe Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Ala Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Val Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Phe Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 1158107PRTArtificial SequenceSynthetic 8Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Val Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Val Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Ile Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Lys Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 1059118PRTArtificial SequenceSynthetic 9Ala Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11510107PRTArtificial SequenceSynthetic 10Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln His Asn Ser Tyr Thr Tyr 85 90 95Lys Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10511118PRTArtificial SequenceSynthetic 11Glu Val Gln Leu Leu Gln Ser Trp Ala Asp Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Asp Lys Ala Ala Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Gly Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Tyr Asp Gly Tyr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11512107PRTArtificial SequenceSynthetic 12Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys His Gln Tyr Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10513118PRTArtificial SequenceSynthetic 13Glu Val Gln Leu Gln Gln Ser Gly Ala Lys Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met Cys Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Ser65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11514107PRTArtificial SequenceSynthetic 14Asp Thr Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Thr Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln His Asn Ser Tyr Ser Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10515117PRTArtificial SequenceSynthetic 15Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Leu Gly Asn Trp Val Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110Leu Thr Val Ser Ser 11516105PRTArtificial SequenceSynthetic 16Asp Thr Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Asp Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Thr Tyr Thr Phe 85 90 95Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10517118PRTArtificial SequenceSynthetic 17Glu Val Gln Leu Gln Gln Phe Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Val Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu His Phe Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Val Arg Gly Gly Asn Phe Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11518107PRTArtificial SequenceSynthetic 18Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Cys Ile Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10519118PRTArtificial SequenceSynthetic 19Glu Val His Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Thr1 5 10 15Ser Leu Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Thr Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Tyr Asn Gly Tyr Ser Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11520107PRTArtificial SequenceSynthetic 20Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Glu Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Arg Tyr Pro Phe 85 90 95Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 10521118PRTArtificial SequenceSynthetic 21Glu Val Gln Leu Leu Gln Ser Trp Ala Asp Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Asp Lys Ala Ala Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Gly Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Tyr Asp Gly Tyr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11522107PRTArtificial SequenceSynthetic 22Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp

Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys His Gln Tyr Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10523118PRTArtificial SequenceSynthetic 23Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Cys Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Val Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Glu Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ser Arg Gly Gly Ser Tyr Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11524107PRTArtificial SequenceSynthetic 24Asp Val Val Met Thr Gln Ser Arg Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ser 20 25 30Val Ala Trp Tyr Gln Gln Lys Leu Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Asp Asn Ser Tyr Pro His 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10525116PRTArtificial SequenceSynthetic 25Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Thr Ser Leu His Trp Ser Leu Asp Ser Trp Gly Gln Gly Thr Thr Leu 100 105 110Thr Val Ser Ser 11526107PRTArtificial SequenceSynthetic 26Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Thr Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ala 20 25 30Val Ala Trp Tyr Gln Gln Gln Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ala Asn Arg Asp Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Met Gln Ser65 70 75 80Glu Asp Leu Ala His Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10527118PRTArtificial SequenceSynthetic 27Asp Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Asn Leu Ser Cys Thr Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Val Asp Pro Glu Asp Gly Glu Thr Lys Tyr Val Pro Lys Phe 50 55 60Leu Asp Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Thr Arg Gly Gly Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11528107PRTArtificial SequenceSynthetic 28Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Phe Leu Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Cys Asn Asn Tyr Arg Leu 85 90 95Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 10529118PRTArtificial SequenceSynthetic 29Glu Val Gln Leu Leu Gln Ser Trp Ala Asp Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Asp Lys Ala Ala Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Gly Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Tyr Asp Gly Tyr Tyr Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11530107PRTArtificial SequenceSynthetic 30Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys His Gln Tyr Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10531119PRTArtificial SequenceSynthetic 31Glu Ala Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Arg Asp Ser 20 25 30Tyr Ile His Trp Val Asn Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Met Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Ser Trp Leu Ala Asp Tyr Ser Ala Met Asp Asn Trp Gly Gln Gly 100 105 110Thr Ser Val Thr Val Ser Ser 11532107PRTArtificial SequenceSynthetic 32Asp Ile Gln Met Thr Gln Ser Ser Ser Ser Phe Ser Val Ser Leu Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Glu Asp Ile Tyr Asn Arg 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Arg Leu Leu Ile 35 40 45Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Lys Asp Tyr Thr Leu Ser Ile Thr Ser Leu Gln Thr65 70 75 80Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Asn Thr Leu Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Met Lys 100 10533119PRTArtificial SequenceSynthetic 33Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ser Ser Gly Phe Asn Ile Lys Asp Ser 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Asn Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg Gly Ser Ser Leu Tyr Ala Val Asp Tyr Trp Gly Gln Gly 100 105 110Thr Ser Val Thr Val Ser Ser 11534107PRTArtificial SequenceSynthetic 34Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ser 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Val Ile 35 40 45Tyr Leu Ala Ser Tyr Arg His Arg Gly Val Pro Ala Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Phe Asn Ile Tyr Pro Trp 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10535118PRTArtificial SequenceSynthetic 35Glu Val Gln Leu Leu Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Arg Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Leu His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Thr Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Thr Arg Gly Gly Asn Tyr Asp Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11536107PRTArtificial SequenceSynthetic 36Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Tyr 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Gly Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Lys Asn Thr Tyr Pro Phe 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Glu 100 10537119PRTArtificial SequenceSynthetic 37Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ser Ser Gly Phe Asn Ile Lys Asp Ser 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Asn Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg Gly Ser Ser Leu Tyr Ala Val Asp Tyr Trp Gly Gln Gly 100 105 110Thr Ser Val Thr Val Ser Ser 11538107PRTArtificial SequenceSynthetic 38Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ser 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Val Ile 35 40 45Tyr Leu Ala Ser Tyr Arg His Arg Gly Val Pro Ala Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Phe Asn Ile Tyr Pro Trp 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10539118PRTArtificial SequenceSynthetic 39Glu Val Gln Leu Gln Gln Ser Gly Ala Asp Leu Val Arg Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Val Lys Thr Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Phe Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Val Arg Gly Gly Asn Phe Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Ser 100 105 110Thr Leu Thr Val Ser Ser 11540107PRTArtificial SequenceSynthetic 40Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Cys Ile Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10541118PRTArtificial SequenceSynthetic 41Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Ser Gly Ala1 5 10 15Ser Val Arg Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Lys Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Val Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Val Tyr65 70 75 80Leu Gln Leu Asn Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Val Arg Gly Gly Asn Tyr Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11542107PRTArtificial SequenceSynthetic 42Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Tyr 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Val Val Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Asn Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Pro65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Asn Tyr Pro Leu 85 90 95Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 10543118PRTArtificial SequenceSynthetic 43Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Leu Ile 35 40

45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Val Lys Ala Thr Ile Thr Ala Asp Ala Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Phe Ser Ser Leu Thr Ser Glu Asp Ala Ala Val Tyr Tyr Cys 85 90 95Val Arg Gly Gly Asn Phe Tyr Tyr Phe Asp Phe Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11544107PRTArtificial SequenceSynthetic 44Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Val 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asp Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Cys Ile Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10545118PRTArtificial SequenceSynthetic 45Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu His Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ser Arg Gly Gly Asn Phe Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Leu Thr Val Ser Ser 11546107PRTArtificial SequenceSynthetic 46Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Leu Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Cys Asn Ser Tyr Ser Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10547118PRTArtificial SequenceSynthetic 47Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Cys Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Val Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Glu Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ser Arg Gly Gly Ser Tyr Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11548107PRTArtificial SequenceSynthetic 48Asp Val Val Met Thr Gln Ser Arg Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ser 20 25 30Val Ala Trp Tyr Gln Gln Lys Leu Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Asp Asn Ser Tyr Pro His 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10549118PRTArtificial SequenceSynthetic 49Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Thr Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ser Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Gly Asp Thr Ala Val Tyr Phe Cys 85 90 95Ala Arg Tyr Asp Gly Tyr Tyr Cys Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11550107PRTArtificial SequenceSynthetic 50Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Ala Asp Leu Ala Ala Tyr Phe Cys Gln Gln Tyr Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys 100 10551117PRTArtificial SequenceSynthetic 51Glu Val Gln Leu Gln Gln Ser Gly Ala Asp Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Thr Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Asn Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Trp Gly His Ser Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110Leu Thr Val Ser Ser 11552105PRTArtificial SequenceSynthetic 52Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Thr 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Thr Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Ser Tyr Thr Phe 85 90 95Gly Gly Gly Thr Lys Leu Glu Met Lys 100 10553119PRTArtificial SequenceSynthetic 53Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Phe Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Thr Gln Lys Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Asp Lys Ala Thr Ile Thr Ala Asp Ser Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Val Asp Thr Ala Val Tyr Phe Cys 85 90 95Ser Arg Glu Gly Ser Phe Thr Gly Trp Phe Pro Tyr Trp Gly Gln Gly 100 105 110Thr Leu Val Ser Val Ser Ala 11554107PRTArtificial SequenceSynthetic 54Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Arg Gln Ser Val Gly Thr Tyr 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Asn Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr His Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10555118PRTArtificial SequenceSynthetic 55Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Thr Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Ala Thr Ile Thr Ser Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Gly Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Tyr Asp Gly Tyr Tyr Cys Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110Thr Leu Thr Val Ser Ser 11556107PRTArtificial SequenceSynthetic 56Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Ala Asp Leu Ala Ala Tyr Phe Cys Gln Gln Tyr Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys 100 10557117PRTArtificial SequenceSynthetic 57Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Ala Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 35 40 45Gly Tyr Ile Ser Tyr Ser Gly Ser Ile Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu65 70 75 80Gln Leu Asn Ser Val Thr Asn Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 85 90 95Arg Ser Gly Gly Met Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110Leu Thr Val Ser Ser 11558107PRTArtificial SequenceSynthetic 58Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly1 5 10 15Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Gly Asn Ile His Asn Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45Tyr Asn Ala Lys Thr Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His Phe Trp Ser Ile Pro Pro 85 90 95Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 10559117PRTArtificial SequenceSynthetic 59Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Ala Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 35 40 45Gly Tyr Ile Ser Tyr Ser Gly Ser Ile Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Ile Ser Ile Thr Arg Ala Thr Ser Lys Asn Gln Tyr Tyr Leu65 70 75 80Gln Leu Asn Ser Val Thr Asn Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 85 90 95Arg Ser Gly Gly Met Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110Leu Thr Val Ser Ser 11560107PRTArtificial SequenceSynthetic 60Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly1 5 10 15Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Gly Asn Ile His Asn Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45Tyr Asn Ala Lys Thr Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His Phe Trp Ser Ile Pro Pro 85 90 95Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 10561117PRTArtificial SequenceSynthetic 61Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Ala Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 35 40 45Gly Tyr Ile Ser Tyr Ser Gly Thr Ile Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu65 70 75 80Gln Leu Asn Ser Val Thr Asn Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 85 90 95Arg Ser Gly Gly Met Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110Leu Thr Val Ser Ser 11562107PRTArtificial SequenceSynthetic 62Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly1 5 10 15Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Gly Asn Ile His Asn Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45Tyr Asn Ala Lys Thr Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His Phe Trp Ser Ile Pro Pro 85 90 95Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 10563117PRTArtificial SequenceSynthetic 63Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Ala Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met 35 40 45Gly Tyr Ile Ser Tyr Ser Gly Asn Thr Asp Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Phe Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Tyr Leu65 70 75 80Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 85 90 95Arg Ser Glu Gly Met Tyr Phe Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110Leu Thr Val Ser Ser 11564107PRTArtificial SequenceSynthetic 64Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly1 5 10 15Glu Thr Val Ser Ile Thr Cys Arg Ala Ser Gly Asn Ile His Asn Tyr 20 25 30Leu Ala Trp Tyr His Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45Tyr Asn Ala Lys Thr Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Asn Ser Leu Gln Pro65

70 75 80Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His Phe Trp Ser Thr Pro Pro 85 90 95Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 10565118PRTArtificial SequenceSynthetic 65Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Phe Gly Gly Leu Thr Ala Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11566107PRTArtificial SequenceSynthetic 66Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45Tyr Ala Thr Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10567118PRTArtificial SequenceSynthetic 67Ala Val Gln Leu Gln Gln Ser Gly Thr Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Ser Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Val His Trp Val Lys Gln Lys Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Val Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Thr Arg Phe Gly Gly Leu Asp Ala Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11568107PRTArtificial SequenceSynthetic 68Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Asn Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Phe Asn Arg Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10569120PRTArtificial SequenceSynthetic 69Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ser1 5 10 15Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Phe Thr Phe Thr Ser His 20 25 30Phe Ile His Trp Ile Lys Gln Gln Pro Gly Asn Gly Leu Lys Trp Ile 35 40 45Gly Trp Ile Tyr Pro Gly Asp Asp Asp Thr Glu Tyr Asn His Lys Phe 50 55 60Asn Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr65 70 75 80Met His Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95Ala Arg Arg Val Glu Tyr Tyr Asn Gly Gly Phe Ala Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser 115 12070107PRTArtificial SequenceSynthetic 70Asp Val Gln Met Thr Gln Ser Pro Ser Tyr Leu Ala Ala Pro Pro Gly1 5 10 15Glu Ser Val Ser Ile Ser Cys Lys Ala Ser Lys Asn Ile Arg Asn Asn 20 25 30Leu Gly Trp Tyr Gln Glu Arg Pro Gly Lys Thr Pro Asn Leu Leu Ile 35 40 45His Ser Gly Ser Thr Leu Gln Ser Gly Ala Pro Ser Arg Phe Ser Gly 50 55 60Gly Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Arg Ser Leu Glu Ser65 70 75 80Glu Asp Ser Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Gln Tyr Pro Leu 85 90 95Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 10571124PRTArtificial SequenceSynthetic 71Gln Val Thr Leu Lys Glu Ser Gly Pro Gly Ile Leu Gln Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Asn Thr Tyr 20 25 30Gly Met Gly Val Gly Trp Ile Arg Gln Pro Ser Gly Lys Gly Leu Glu 35 40 45Trp Leu Ala Asn Ile Trp Trp Asp Asp Asp Lys Tyr Tyr Asn Pro Ser 50 55 60Leu Lys Asn Arg Leu Thr Ile Ser Lys Asp Thr Ser Asn Asn Gln Ala65 70 75 80Phe Leu Lys Ile Thr Asn Val Asp Thr Ala Asp Thr Ala Thr Tyr Phe 85 90 95Cys Ala Arg His Pro Leu Pro Gly Tyr Lys Asp Asn Tyr Val Val Asp 100 105 110Ala Trp Gly Gln Gly Ala Ser Val Thr Val Ser Ser 115 12072112PRTArtificial SequenceSynthetic 72Asp Val Val Leu Thr Gln Thr Pro Gly Ser Leu Ser Val Thr Leu Gly1 5 10 15Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Glu Tyr Ser 20 25 30Asp Gln Tyr Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Ser Gly Gln Ser 35 40 45Pro Gln Leu Leu Ile Tyr Gly Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Pro Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Ala 85 90 95Thr His Asp Pro Tyr Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 11073118PRTArtificial SequenceSynthetic 73Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Thr Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Val Val Tyr Tyr Cys 85 90 95Gly Arg Tyr His Gly Tyr Trp Ala Leu Asp Tyr Trp Gly Gln Gly Thr 100 105 110Ser Val Thr Val Ser Ser 11574107PRTArtificial SequenceSynthetic 74Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Val Thr Cys Lys Thr Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Ile Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys His Gln Tyr Phe Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gly Gly Thr Leu Leu Glu Ile Lys 100 10575117PRTArtificial SequenceSynthetic 75Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Ser Ala Ser Gly Phe Asn Ile Lys Asp Cys 20 25 30Tyr Ile His Trp Val Lys Gln Arg Thr Glu Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Ala Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr65 70 75 80Leu Gln Leu Asn Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg His Cys Asn Phe Leu Tyr Phe Asp Tyr Trp Gly Gln Gly Ser 100 105 110Thr Leu Thr Val Ser 11576107PRTArtificial SequenceSynthetic 76Asp Ile Val Met Thr Gln Ser Gln Lys Ser Met Ser Thr Ser Leu Gly1 5 10 15Asp Arg Val Thr Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ile 20 25 30Val Ala Trp Tyr Gln Leu Lys Pro Gly Gln Ser Pro Lys Thr Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Ser Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser65 70 75 80Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr Asn Asn Tyr Pro Val 85 90 95Ile Phe Gly Ser Gly Thr Lys Leu Glu Ile Arg 100 105775PRTArtificial SequenceSynthetic 77Asp Tyr Tyr Met His1 57817PRTArtificial SequenceSynthetic 78Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Gly7917PRTArtificial SequenceSynthetic 79Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Gly809PRTArtificial SequenceSynthetic 80Gly Gly Asn Phe Tyr Val Met Asp Tyr1 58111PRTArtificial SequenceSynthetic 81Lys Ala Ser Gln Asn Val Gly Thr Asn Val Val1 5 10827PRTArtificial SequenceSynthetic 82Ser Ala Ser Tyr Arg Val Ser1 5839PRTArtificial SequenceSynthetic 83Gln Gln Lys Asn Asn Tyr Pro Tyr Thr1 584118PRTArtificial SequenceSynthetic 84Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Phe Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11585118PRTArtificial SequenceSynthetic 85Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Ala Asp Thr Ser Thr Ser Thr Val Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Phe Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11586118PRTArtificial SequenceSynthetic 86Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Val Tyr65 70 75 80Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Phe Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11587118PRTArtificial SequenceSynthetic 87Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Lys Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Ala Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75 80Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Asn Phe Tyr Val Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11588107PRTArtificial SequenceSynthetic 88Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Val Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10589107PRTArtificial SequenceSynthetic 89Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Val Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Val Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Ile Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10590107PRTArtificial SequenceSynthetic 90Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Val Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Val Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Ile Ile Ser Ser Val Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Lys Asn Asn Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10591107PRTArtificial SequenceSynthetic 91Asp Ile Gln Met Thr Gln Ser Pro Ser Phe Leu Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Thr Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30Val Val Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Val Leu Ile 35 40 45Tyr Ser Ala Ser Tyr Arg Val Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Ile Ile Ser Ser Val Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Lys Asn Asn Tyr Pro Tyr 85 90

95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105925PRTArtificial SequenceSynthetic 92Asp Tyr Tyr Ile His1 59317PRTArtificial SequenceSynthetic 93Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Gly9417PRTArtificial SequenceSynthetic 94Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Gly959PRTArtificial SequenceSynthetic 95Tyr His Gly Tyr Trp Ala Leu Asp Tyr1 59611PRTArtificial SequenceSynthetic 96Lys Thr Ser Gln Asn Val Gly Thr Asn Val Ala1 5 10977PRTArtificial SequenceSynthetic 97Ser Thr Ser Tyr Arg Tyr Ser1 5989PRTArtificial SequenceSynthetic 98His Gln Tyr Phe Ser Tyr Pro Tyr Thr1 599118PRTArtificial SequenceSynthetic 99Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Val Val Tyr Tyr Cys 85 90 95Ala Arg Tyr His Gly Tyr Trp Ala Leu Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 115100118PRTArtificial SequenceSynthetic 100Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Ala Asp Thr Ser Ile Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Val Val Tyr Tyr Cys 85 90 95Ala Arg Tyr His Gly Tyr Trp Ala Leu Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 115101118PRTArtificial SequenceSynthetic 101Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Ala Asp Thr Ser Ile Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Val Val Tyr Tyr Cys 85 90 95Gly Arg Tyr His Gly Tyr Trp Ala Leu Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 115102118PRTArtificial SequenceSynthetic 102Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10 15Ser Val Lys Leu Ser Cys Lys Gly Ser Gly Phe Asn Ile Lys Asp Tyr 20 25 30Tyr Ile His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45Gly Arg Ile Asp Pro Glu Asp Ala Glu Thr Lys Tyr Ala Pro Lys Phe 50 55 60Gln Gly Arg Ala Thr Ile Thr Ala Asp Thr Ser Ile Ser Thr Ala Tyr65 70 75 80Leu Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Val Val Tyr Tyr Cys 85 90 95Gly Arg Tyr His Gly Tyr Trp Ala Leu Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 115103107PRTArtificial SequenceSynthetic 103Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Thr Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Phe Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105104107PRTArtificial SequenceSynthetic 104Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Thr Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Phe Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105105107PRTArtificial SequenceSynthetic 105Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Lys Thr Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Phe Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Pro Gly Thr Lys Leu Asp Ile Lys 100 105106107PRTArtificial SequenceSynthetic 106Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Thr Val Thr Cys Lys Thr Ser Gln Asn Val Gly Thr Asn 20 25 30Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Pro Leu Ile 35 40 45Tyr Ser Thr Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Phe Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Pro Gly Thr Lys Leu Asp Ile Lys 100 10510717PRTArtificial SequenceSynthetic 107Arg Ile Asp Pro Asp Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Gly1089PRTArtificial SequenceSynthetic 108Gly Gly Asn Tyr Tyr Val Met Asp Asn1 510911PRTArtificial SequenceSynthetic 109Lys Ala Ser Gln Asn Val Gly Thr Asn Val Ala1 5 101107PRTArtificial SequenceSynthetic 110Thr Ala Ser Tyr Arg Tyr Ser1 51119PRTArtificial SequenceSynthetic 111Gln Gln Tyr Phe Ser Tyr Pro His Thr1 51125PRTArtificial SequenceSynthetic 112Asp Ser Tyr Met His1 511317PRTArtificial SequenceSynthetic 113Arg Ile Asp Pro Glu Asp Gly Glu Thr Asn Tyr Ala Pro Lys Phe Gln1 5 10 15Gly11410PRTArtificial SequenceSynthetic 114Arg Val Ser Ser Leu Tyr Ala Met Asp Tyr1 5 101157PRTArtificial SequenceSynthetic 115Leu Ala Ser Tyr Arg Tyr Ser1 51169PRTArtificial SequenceSynthetic 116Gln Gln Tyr Asn Asn Tyr Pro Trp Thr1 51177PRTArtificial SequenceSynthetic 117Ser Ala Ser Tyr Arg Tyr Ser1 51189PRTArtificial SequenceSynthetic 118Gln Gln Lys Asn Ser Tyr Pro Tyr Thr1 51199PRTArtificial SequenceSynthetic 119Gly Gly Asn Tyr Tyr Ala Met Asp Tyr1 51209PRTArtificial SequenceSynthetic 120Gln Gln His Asn Ser Tyr Thr Tyr Lys1 512117PRTArtificial SequenceSynthetic 121Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Asp1229PRTArtificial SequenceSynthetic 122Tyr Asp Gly Tyr Tyr Gly Phe Asp Tyr1 51239PRTArtificial SequenceSynthetic 123His Gln Tyr Asn Asn Tyr Pro Tyr Thr1 51245PRTArtificial SequenceSynthetic 124Asp Tyr Tyr Met Cys1 51257PRTArtificial SequenceSynthetic 125Ser Ala Thr Tyr Arg Tyr Ser1 51269PRTArtificial SequenceSynthetic 126Gln Gln His Asn Ser Tyr Ser Tyr Thr1 51278PRTArtificial SequenceSynthetic 127Leu Gly Asn Trp Val Phe Asp Tyr1 512811PRTArtificial SequenceSynthetic 128Lys Ala Ser Gln Asn Val Gly Thr Asn Val Asp1 5 101297PRTArtificial SequenceSynthetic 129Gln Gln Tyr Asn Thr Tyr Thr1 513017PRTArtificial SequenceSynthetic 130Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Val1319PRTArtificial SequenceSynthetic 131Gly Gly Asn Phe Tyr Tyr Phe Asp Tyr1 51329PRTArtificial SequenceSynthetic 132Gln Gln Cys Ile Asn Tyr Pro Tyr Thr1 513317PRTArtificial SequenceSynthetic 133Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Thr Phe Gln1 5 10 15Gly1349PRTArtificial SequenceSynthetic 134Tyr Asn Gly Tyr Ser Gly Phe Asp Tyr1 51359PRTArtificial SequenceSynthetic 135Gln Gln Tyr Asn Arg Tyr Pro Phe Thr1 513617PRTArtificial SequenceSynthetic 136Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Val Pro Lys Phe Gln1 5 10 15Gly1379PRTArtificial SequenceSynthetic 137Gly Gly Ser Tyr Tyr Val Met Asp Tyr1 513811PRTArtificial SequenceSynthetic 138Lys Ala Ser Gln Asn Val Gly Thr Ser Val Ala1 5 101399PRTArtificial SequenceSynthetic 139Gln Gln Asp Asn Ser Tyr Pro His Thr1 51407PRTArtificial SequenceSynthetic 140Leu His Trp Ser Leu Asp Ser1 514111PRTArtificial SequenceSynthetic 141Lys Ala Ser Gln Asn Val Gly Thr Ala Val Ala1 5 101427PRTArtificial SequenceSynthetic 142Ser Thr Ala Asn Arg Asp Thr1 51439PRTArtificial SequenceSynthetic 143Gln Gln Tyr Ser Ser Tyr Pro Tyr Thr1 514417PRTArtificial SequenceSynthetic 144Arg Val Asp Pro Glu Asp Gly Glu Thr Lys Tyr Val Pro Lys Phe Leu1 5 10 15Asp1459PRTArtificial SequenceSynthetic 145Gln Gln Cys Asn Asn Tyr Arg Leu Thr1 51465PRTArtificial SequenceSynthetic 146Asp Ser Tyr Ile His1 514710PRTArtificial SequenceSynthetic 147Trp Leu Ala Asp Tyr Ser Ala Met Asp Asn1 5 1014811PRTArtificial SequenceSynthetic 148Lys Ala Ser Glu Asp Ile Tyr Asn Arg Leu Ala1 5 101497PRTArtificial SequenceSynthetic 149Gly Ala Thr Ser Leu Glu Thr1 51509PRTArtificial SequenceSynthetic 150Gln Gln Tyr Trp Asn Thr Leu Tyr Thr1 515110PRTArtificial SequenceSynthetic 151Arg Gly Ser Ser Leu Tyr Ala Val Asp Tyr1 5 101527PRTArtificial SequenceSynthetic 152Leu Ala Ser Tyr Arg His Arg1 51539PRTArtificial SequenceSynthetic 153Gln Gln Phe Asn Ile Tyr Pro Trp Thr1 51545PRTArtificial SequenceSynthetic 154Asp Tyr Tyr Leu His1 51559PRTArtificial SequenceSynthetic 155Gly Gly Asn Tyr Asp Val Met Asp Tyr1 515611PRTArtificial SequenceSynthetic 156Lys Ala Ser Gln Asn Val Gly Thr Tyr Val Val1 5 101579PRTArtificial SequenceSynthetic 157Gln Gln Lys Asn Thr Tyr Pro Phe Thr1 51589PRTArtificial SequenceSynthetic 158Gly Gly Asn Tyr Tyr Val Met Asp Tyr1 515911PRTArtificial SequenceSynthetic 159Lys Ala Ser Gln Asn Val Gly Thr Tyr Val Ala1 5 101607PRTArtificial SequenceSynthetic 160Ser Ala Ser Tyr Arg Tyr Asn1 51619PRTArtificial SequenceSynthetic 161Gln Gln Tyr Asn Asn Tyr Pro Leu Thr1 51629PRTArtificial SequenceSynthetic 162Gly Gly Asn Phe Tyr Tyr Phe Asp Phe1 51639PRTArtificial SequenceSynthetic 163Gln Gln Cys Asn Ser Tyr Ser Tyr Thr1 51649PRTArtificial SequenceSynthetic 164Tyr Asp Gly Tyr Tyr Cys Phe Asp Tyr1 51659PRTArtificial SequenceSynthetic 165Gln Gln Tyr Asn Asn Tyr Pro Tyr Thr1 51668PRTArtificial SequenceSynthetic 166Asp Trp Gly His Ser Phe Asp Tyr1 516711PRTArtificial SequenceSynthetic 167Lys Ala Ser Gln Asn Val Gly Thr Thr Val Ala1 5 101687PRTArtificial SequenceSynthetic 168Gln Gln Tyr Asn Ser Tyr Thr1 516910PRTArtificial SequenceSynthetic 169Glu Gly Ser Phe Thr Gly Trp Phe Pro Tyr1 5 1017011PRTArtificial SequenceSynthetic 170Lys Ala Arg Gln Ser Val Gly Thr Tyr Val Ala1 5 101717PRTArtificial SequenceSynthetic 171Ser Thr Ser Tyr Arg Tyr Asn1 51729PRTArtificial SequenceSynthetic 172Gln Gln Tyr His Ser Tyr Pro Tyr Thr1 51735PRTArtificial SequenceSynthetic 173Ser Asp Tyr Trp Asn1 517416PRTArtificial SequenceSynthetic 174Tyr Ile Ser Tyr Ser Gly Ser Ile Tyr Tyr Asn Pro Ser Leu Lys Ser1 5 10 151759PRTArtificial SequenceSynthetic 175Ser Gly Gly Met Tyr Tyr Phe Asp Tyr1 517611PRTArtificial SequenceSynthetic 176Arg Ala Ser Gly Asn Ile His Asn Tyr Leu Ala1 5 101777PRTArtificial SequenceSynthetic 177Asn Ala Lys Thr Leu Glu Asp1 51789PRTArtificial SequenceSynthetic 178Gln His Phe Trp Ser Ile Pro Pro Thr1 517916PRTArtificial SequenceSynthetic 179Tyr Ile Ser Tyr Ser Gly Thr Ile Tyr Tyr Asn Pro Ser Leu Lys Ser1 5 10 1518016PRTArtificial SequenceSynthetic 180Tyr Ile Ser Tyr Ser Gly Asn Thr Asp Tyr Asn Pro Ser Leu Lys Ser1 5 10 151819PRTArtificial SequenceSynthetic 181Ser Glu Gly Met Tyr Phe Phe Asp Tyr1 51827PRTArtificial SequenceSynthetic 182Asn Ala Lys Thr Leu Ala Asp1 51839PRTArtificial SequenceSynthetic 183Gln His Phe Trp Ser Thr Pro Pro Thr1 51849PRTArtificial SequenceSynthetic 184Phe Gly Gly Leu Thr Ala Met Asp Tyr1 51857PRTArtificial SequenceSynthetic 185Ala Thr Ser Tyr Arg Tyr Ser1 51865PRTArtificial SequenceSynthetic 186Asp Tyr Tyr Val His1 51879PRTArtificial SequenceSynthetic 187Phe Gly Gly Leu Asp Ala Met Asp Tyr1 51889PRTArtificial SequenceSynthetic 188Gln Gln Phe Asn Arg Tyr Pro Tyr Thr1 51895PRTArtificial SequenceSynthetic 189Ser His Phe Ile His1 519017PRTArtificial SequenceSynthetic 190Trp Ile Tyr Pro Gly Asp Asp Asp Thr Glu Tyr Asn His Lys Phe Asn1 5 10 15Gly19111PRTArtificial SequenceSynthetic 191Arg Val Glu Tyr Tyr Asn Gly Gly Phe Ala Tyr1 5 1019211PRTArtificial SequenceSynthetic 192Lys Ala Ser Lys Asn Ile Arg Asn Asn Leu Gly1 5 101937PRTArtificial SequenceSynthetic 193Ser Gly Ser Thr Leu Gln Ser1 51949PRTArtificial SequenceSynthetic 194Gln Gln Tyr Asp Gln Tyr Pro Leu Thr1 51957PRTArtificial SequenceSynthetic 195Thr Tyr Gly Met Gly Val Gly1 519616PRTArtificial SequenceSynthetic 196Asn Ile Trp Trp Asp Asp Asp Lys Tyr Tyr Asn Pro Ser Leu Lys Asn1 5 10 1519714PRTArtificial SequenceSynthetic 197His Pro Leu Pro Gly Tyr Lys Asp Asn Tyr Val Val Asp Ala1 5 1019816PRTArtificial SequenceSynthetic 198Arg Ser Ser Gln Ser Leu Glu Tyr Ser Asp Gln Tyr Thr Tyr Leu Glu1 5 10 151997PRTArtificial SequenceSynthetic 199Gly Val Ser Asn Arg Phe Ser1 52009PRTArtificial SequenceSynthetic 200Phe Gln Ala Thr His Asp Pro Tyr Thr1 52015PRTArtificial SequenceSynthetic 201Asp Cys Tyr Ile His1 520217PRTArtificial SequenceSynthetic 202Arg Ile Asp Pro Glu Asp Gly Glu Thr Lys Tyr Ala Pro Lys Phe Gln1 5 10 15Ala2039PRTArtificial SequenceSynthetic 203His Cys Asn Phe

Leu Tyr Phe Asp Tyr1 520411PRTArtificial SequenceSynthetic 204Lys Ala Ser Gln Asn Val Gly Thr Ile Val Ala1 5 102057PRTArtificial SequenceSynthetic 205Ser Ala Ser Tyr Arg Ser Ser1 52069PRTArtificial SequenceSynthetic 206Gln Gln Tyr Asn Asn Tyr Pro Val Ile1 5

Claims

1. An antibody or fragment thereof having specificity to a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein, wherein the antibody or fragment thereof comprises a heavy chain variable region comprising heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and a light chain variable region comprising light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises the amino acid sequence of DYYIH (SEQ ID NO: 92), the HCDR2 comprises the amino acid sequence of RIDPEDGETKYAPKFQG (SEQ ID NO: 93) or RIDPEDAETKYAPKFQG (SEQ ID NO:94), the HCDR3 comprises the amino acid sequence of YHGYWALDY (SEQ ID NO: 95), the LCDR1 comprises the amino acid sequence of KTSQNVGTNVA (SEQ ID NO: 96), the LCDR2 comprises the amino acid sequence of STSYRYS (SEQ ID NO: 97), and the LCDR3 comprises the amino acid sequence of HQYFSYPYT (SEQ ID NO: 98).

2. The antibody or fragment thereof of claim 1, wherein the antibody is a chimeric antibody or a humanized antibody.

3. The antibody or fragment thereof of claim 1, which is humanized and wherein the heavy chain variable region comprises one or more back mutations selected from the group consisting of 20L, 24G, 38K, 48I, 68A, 70I, 72A, 81L, and 97G, according to Kabat numbering, and combinations thereof.

4. The antibody or fragment thereof of claim 1, which is humanized and wherein the light chain variable region comprises one or more back mutations selected from the group consisting of 13T, 21V, 36Y, 46P, 78V, and 104L, according to Kabat numbering, and combinations thereof.

5. The antibody or fragment thereof of claim 1, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 73, and 99-102.

6. The antibody or fragment thereof of claim 1, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 74, and 103-106.

7. The antibody or fragment thereof of claim 1, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 100 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 105.

8. A bifunctional molecule, comprising a first antigen-binding portion having specificity to a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein and a second portion having specificity to a second protein, wherein the first antigen-binding portion comprises an antibody fragment of claim 1.

9. The bifunctional molecule of claim 8, wherein the second portion comprises peptide edratide (hCDR1) or TACI-Ig.

10. The bifunctional molecule of claim 8, wherein the second portion is an antigen-binding fragment having specificity to a protein selected from the group consisting of BAFF, CD20, CD22, CTLA4, IL6, CXCL13 and C5.

11. One or more polynucleotide encoding the antibody or fragment thereof of claim 1.

12. A method of suppressing an immune response or treating an autoimmune disease or disorder in a patient in need thereof, comprising administering to the patient the antibody or fragment thereof of claim 1.

13. The method of claim 12, for treating an autoimmune disease or disorder.

14. The method of claim 13, wherein the autoimmune disease or disorder is selected from the group consisting of type 1 diabetes, rheumatoid arthritis (RA), psoriasis/psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus (lupus), inflammatory bowel disease, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, and celiac disease.

15. A method of detecting expression of a human interferon alpha and beta receptor subunit 1 (IFNAR1) protein in a sample, comprising contacting the sample with the antibody or fragment thereof of claim 1, and detecting the binding which indicates expression of IFNAR1 in the sample.