ANTI-MESOTHELIN ANTIBODIES AND USES THEREOF

Abstract

The present invention provides recombinant antigen-binding regions and antibodies and functional fragments containing such antigen-binding regions that are specific for the membrane-anchored, 40 kDa mesothelin polypeptide, which is overexpressed in several tumors, such as pancreatic and ovarian tumors, mesothelioma and lung cancer cells. These antibodies, accordingly, can be used to treat these and other disorders and conditions. Antibodies of the invention also can be used in the diagnostics field, as well as for further investigating the role of mesothelin in the progression of disorders associated with cancer. The invention also provides nucleic acid sequences encoding the foregoing antibodies, vectors containing the same, pharmaceutical compositions and kits with instructions for use.

Description

[0001] The present invention provides recombinant antigen-binding regions and antibodies and functional fragments containing such antigen-binding regions that are specific for the membrane-anchored, 40 kDa mesothelin polypeptide, which is overexpressed in several tumors, such as pancreatic and ovarian tumors, mesothelioma and lung cancer cells. These antibodies, accordingly, can be used to treat these and other disorders and conditions. Antibodies of the invention also can be used in the diagnostics field, as well as for further investigating the role of mesothelin in the progression of disorders associated with cancer. The invention also provides nucleic acid sequences encoding the foregoing antibodies, vectors containing the same, pharmaceutical compositions and kits with instructions for use.

BACKGROUND OF THE INVENTION

[0002] Antibody-based therapy is proving very effective in the treatment of various cancers, including solid tumors. For example, HERCEPTIN® has been used successfully to treat breast cancer. Central to the development of a successful antibody-based therapy is isolation of antibodies against cell-surface proteins found to be preferentially expressed on tumor cells. The mesothelin precursor polypeptide is a glycophosphatidylinositol (GPI)-anchored, glycosylated cell surface protein that is proteolytically cleaved to a 30 kDa N-terminal secreted polypeptide and a 40 kDa, C-terminal polypeptide, which predominantly occurs in the membrane-bound, GPI-anchored form (Chang, K. and I. Pastan, Proc. Natl. Acad. Sci. USA, (1996) 93(1):136), and which is named mesothelin herein. Mesothelin is preferentially expressed by certain tumor cells, particularly mesothelioma cells, pancreatic tumor cells and ovarian carcinoma cells, while its expression is limited in normal tissue, making it an ideal target for tumor therapy (Argani, P. et al., Clin. Cancer Res. (2001) 7(12): 3862; Hassan, R., et al., Clin. Cancer Res. (2004) 10(12 Pt 1):3937). The function of mesothelin is unknown, and no apparent reproductive, hematologic, or anatomic abnormalities were observed in mice deficient in mesothelin gene expression (Bera, T. K. and I. Pastan, Mol. Cell. Biol. (2000) 20(8):2902).

[0003] Antibody-based, targeted therapy against mesothelin-expressing cancer cells has been proposed for the treatment of lung, ovarian and pancreatic cancer. Mab K1 was the first antibody to membrane-bound mesothelin polypeptide which was described (Chang, K., et al., Int. J. Cancer, (1992) 50(3):373). Mab K1 was generated by immunizing mice. Due to low affinity and poor internalization rates of the antibody, an immunotoxin consisting of Mab K1 linked to a chemically modified truncated form of Pseudomonas exotoxin A was not considered suitable for clinical development (Hassan, R., et al., J. Immunother. (2000) 23(4):473; Hassan, R., et al., Clin. Cancer Res. (2004) 10(12 Pt 1): 3937). Subsequently, single-chain antibodies with higher affinities were developed, including SS1-(dsFv)-PE38, which showed killing activity of tumor cells in vitro (Hassan, R., et al., Clin. Cancer Res. (2002) 8(11): 3520) as well as potency in a murine model of human mesothelin-expressing tumors (Fan, D., et al., Mol. Cancer Ther. (2002) 1(8): 595). These data validate mesothelin as a suitable target for immunotherapy of multiple cancers. However, in clinical trials, SS1-(dsFv)-PE38 was immunogenic, preventing a second administration for the majority of patients. Furthermore, SS1-(dsFv)-PE38 has been shown to have a fast blood clearance and attempts are being reported to increase the molecular weight by pegylating the fusion protein (Filpula, D., et al., Bioconjugate Chem. (2007) 18(3): 773).

[0004] MS-1, MS-2 and MS-3 are mesothelin-binding antibodies which elicit immune effector activity at the cell surface due to their human IgG1 isotype and internalize into mesothelin expressing cells (WO 2006/099141 A2). One of these antibodies, the unconjugated IgG anti-mesothelin antibody MOR Ab 009 is currently being tested in a clinical trial for therapeutic effects in the treatment of pancreatic cancer.

[0005] The predictive value of xenograft murine cancer models for clinical outcome of immunotoxin cancer therapy is often limited by a lack of cross-reactivity of the therapeutic antibodies with their murine homologues, which leads to reduced unspecific binding to normal tissue. On the other hand, neutralizing anti-mouse Fv antibodies which are formed in patients being treated with murine or chimeric antibodies may result in either dose-limiting toxicity or diminished therapeutic potency. Thus, to fully exploit the potential of specific mesothelin expression in cancer therapy, targeting antibodies are required which combine the advantages of increased affinities and reduced dissociation rates with a fully human variable chain format, and with murine cross-reactivity.

[0006] A further necessary feature of novel antibodies is invariant affinity to different cancer cell lines expressing mesothelin on their surface. Mesothelin is a highly variable protein, undergoing post-translational proteolytic digestion as well as glycosylation at multiple sites (Hassan, R., et al., Clin. Cancer Res. (2004) 10(12 Pt 1): 3937). Variability extends to the transcriptional level, since three different splice variants have been detected, although transcript variant 1 (NM 005823) seems to represent the major species present in tumor cell lines tested so far (Muminova, Z. E., et al., BMC Cancer (2004) 4:19; Hellstrom, I., et al., Cancer Epidemiol. Biomarkers Prev. (2006) 15(5):1014). Thus, effective anti-mesothelin antibodies must bind to an epitope invariantly presented by tumor cells from different patients, independently of individual variance including, but not restricted to, variances in glycosylation patterns, which leads to the expression of different forms of mesothelin.

[0007] Provided herein are antibodies, antigen-binding antibody fragments thereof, or variants thereof, that bind to mesothelin with high and invariant affinity, internalize efficiently, and that are preferably cross-reactive to mesothelin from another species. Also provided are antibody-based therapies for cancer, in particular for mesothelin expressing tumors, for example pancreatic, ovarian, or lung cancer, using antibodies, antigen-binding antibody fragments thereof, or variants thereof, that facilitate delivery of therapeutically active agents to cancer cells.

SUMMARY OF THE INVENTION

[0008] It is an object of the invention to provide human and humanized antibodies, or antigen-binding antibody fragments thereof, or variants thereof, that are highly selective for the 40 kDa, C-terminal extracellular part of the mesothelin precursor polypeptide, and which may be employed in methods for detection of mesothelin expression, which is associated with disease states such as cancer of the pancreas, ovary, and lung, and in the treatment of such disease states. Toward these ends, it is an object of the invention to provide isolated human antibodies, or antigen binding antibody fragments thereof, that specifically bind to an epitope present in the mesothelin polypeptide (SEQ ID NO:370), which is invariantly presented by mesothelin-expressing cancer cell lines, and which is bound by these antibodies with comparable affinities. As used herein, the term `invariant presentation` of the epitope refers to the presence of an epitope recognized by a particular antibody on a broad range of mesothelin expressing tumor cell lines which express different forms of mesothelin. As used herein, different `forms` of mesothelin include, but are not restricted to, different glycoforms, different isoforms or mesothelin polypeptides which undergo different translational and posttranslational modifications. As used herein, the term `comparable affinities` refers to half-maximal antibody potency (EC₅₀) values obtained by Scatchard Analysis of FACS data of antibody binding to cells expressing different forms of mesothelin, which do not differ by more than factor 10, or, preferably, factor 5, or, even preferably, factor 2.

[0009] It is another object of the invention to provide antibodies, or antigen-binding antibody fragments thereof, or variants thereof that are safe for human administration.

[0010] It is another object of the invention to provide antibodies, or antigen-binding antibody fragments thereof, or variants thereof, which bind to human mesothelin and are cross-reactive to mesothelin of another species. Preferably said other species is a rodent, such as for example mouse or rat. Most preferably the antibodies, or antigen-binding antibody fragments thereof, or variants thereof bind to human mesothelin and are cross-reactive to murine mesothelin.

[0011] It is another object of the invention to provide antibodies, or antigen-binding antibody fragments thereof, or variants thereof, which invariantly bind to different mesothelin-expressing cell lines with comparable affinity. As used herein, the term `invariant binding` of a particular antibody to mesothelin refers to its ability to bind to mesothelin on a broad range of mesothelin-expressing cancer cell lines which express different forms of mesothelin. Invariant binding may be caused by, but is not restricted to, the fact that antibodies, or antigen-binding antibody fragments thereof, or variants thereof, recognize an epitope of mesothelin that is not masked by another extracellular antigen, such as cancer antigen 125 (CA125), which interacts with mesothelin.

[0012] It is another object of the invention to provide antibodies or variants thereof, which bind to different mesothelin-expressing cancer cells or tumor cells and elicit immune effector activity (e.g. ADCC or CDC) against mesothelin-expressing cancer cells, by using one or more antibodies or variants thereof, of the invention.

[0013] It is another object of the invention to provide antibodies, or antigen-binding antibody fragments thereof, or variants thereof, which are internalized following binding to a mesothelin expressing cell. It is also an object of the present invention to provide methods for treating disease by delivering cytotoxic drugs or drug-releasing enzymes to mesothelin-expressing cancer cells, by using one or more antibodies, or antigen-binding antibody fragments thereof, or variants thereof, of the invention.

[0014] It is another object of the invention to provide antibodies which constitute a tool for diagnosis of malignant or dysplastic conditions in which mesothelin expression is elevated compared to normal tissue. Provided are anti-mesothelin antibodies conjugated to a detectable marker. Preferred markers are a radiolabel, an enzyme, a chromophore or a fluorescer.

[0015] The invention is also related to polynucleotides encoding the antibodies of the invention, cells expressing the antibodies of the invention, methods for producing the antibodies of the invention, methods for inhibiting the growth of dysplastic cells using the antibodies, and methods for treating and detecting cancer using the antibodies.

[0016] The invention provides antibodies that are distinguished from Mab K1, SS1, MS-1, MS-2 and MS-3 in that they a) invariantly bind to mesothelin b) are cross-reactive to murine mesothelin c) bind to mesothelin with lower affinities d) internalize efficiently into mesothelin-expressing cells, and e) contain human variable regions.

[0017] These and other objects of the invention are more fully described herein.

[0018] In one aspect, the invention provides an isolated antibody or functional antibody fragment that contains an antigen-binding region that is specific for an epitope of the 40 kDa mesothelin polypeptide.

[0019] Such an antibody or functional fragment thereof may contain an antigen-binding region that contains an H-CDR3 region depicted in SEQ ID NO: 67-98; the antigen-binding region may further include an H-CDR2 region depicted in SEQ ID NO:31-66; and the antigen-binding region also may contain an H-CDR1 region depicted in SEQ ID NO:1-30. Such a mesothelin-specific antibody of the invention may contain an antigen-binding region that contains an L-CDR3 region depicted in SEQ ID NO:160-197; the antigen-binding region may further include an L-CDR1 region depicted in SEQ ID NO:99-128; and the antigen-binding region also may contain an L-CDR2 region depicted in SEQ ID NO:129-159.

[0020] Peptide variants of the sequences disclosed herein are also embraced by the present invention. Accordingly, the invention includes anti-mesothelin antibodies having a heavy chain amino acid sequence with: at least 60 percent sequence identity in the CDR regions with the CDR regions depicted in SEQ ID NO:1-197; and/or at least 80 percent sequence homology in the CDR regions with the CDR regions depicted in SEQ ID NO: 1-197. Further included are anti-mesothelin antibodies having a light chain amino acid sequence with: at least 60 percent sequence identity in the CDR regions with the CDR regions depicted in SEQ ID NO: 1-197; and/or at least 80 percent sequence homology in the CDR regions with the CDR regions depicted in SEQ ID NO: 1-197.

[0021] An antibody of the invention may be an IgG (e.g., IgG₁), while an antibody fragment may be a Fab or scFv, for example. An inventive antibody fragment, accordingly, may be, or may contain, an antigen-binding region that behaves in one or more ways as described herein.

[0022] The invention also is related to isolated nucleic acid sequences, each of which can encode an antigen-binding region of a human antibody or functional fragment thereof that is specific for an epitope of mesothelin. Such a nucleic acid sequence may encode a variable heavy chain of an antibody and include a sequence selected from the group consisting of SEQ ID NOS 284-326: or a nucleic acid sequence that hybridizes under high stringency conditions to the complementary strand of SEQ ID NO: 284-326. The nucleic acid might encode a variable light chain of an isolated antibody or functional fragment thereof, and may contain a sequence selected from the group consisting of SEQ ID NOS: 327-369, or a nucleic acid sequence that hybridizes under high stringency conditions to the complementary strand of SEQ ID NO: 327-369.

[0023] Nucleic acids of the invention are suitable for recombinant production. Thus, the invention also relates to vectors and host cells containing a nucleic acid sequence of the invention.

[0024] Compositions of the invention may be used for therapeutic or prophylactic applications. The invention, therefore, includes a pharmaceutical composition containing an inventive antibody (or functional antibody fragment) and a pharmaceutically acceptable carrier or excipient therefor. In a related aspect, the invention provides a method for treating a disorder or condition associated with the undesired presence of mesothelin expressing cells. Such method contains the steps of administering to a subject in need thereof an effective amount of the pharmaceutical composition that contains an inventive antibody as described or contemplated herein.

[0025] The invention also provides instructions for using the antibody library to isolate one or more members of such library that binds specifically and invariantly to mesothelin.

DESCRIPTION OF THE FIGURES

[0026] FIG. 1 shows anti-mesothelin antibody epitope grouping by Biacore pairwise binding analysis. Competitive binding of pairs of antibodies was determined by immobilizing one antibody to the sensor chip, binding soluble mesothelin to this antibody and immediately binding a second antibody to mesothelin. Pairs of antibodies which recognize the same or overlapping epitopes on mesothelin cannot bind simultaneously. All combinations of antibody pairs were tested. Representative data for MF-T are shown (A). Panel B depicts the relative positions of epitopes of seven anti-mesothelin antibodies, in which competition is depicted by overlapping circles.

[0027] FIG. 2 shows different forms of mesothelin recognized by antibodies of the invention. 1. and 2.: MF-J binding to mesothelin in OVCAR-3 cell extracts; 3. and 4.: MF-J binding to mesothelin in CHO-A9 cell extracts; 5. MF-J binding to mesothelin in NCI-H226 cell extracts; 6. MF-J binding to recombinant, deglycosylated mesothelin; 7. MOR06635 binding to OVCAR-3 cell extracts; and 8. MOR06635 binding to NCI-H226 cell extracts.

[0028] FIG. 3 shows that cancer antigen 125 (CA125) binds mesothelin when it is bound to a subset of mesothelin antibodies including MOR06640 and MF-T, while other antibodies, such as MF-226, compete with CA125 for mesothelin binding. Data shown are relative light units (RLU) detected by SECTOR Light Imager (Meso Scale Discovery). Plates were coated with the mesothelin antibody depicted. Mesothelin was added at the concentrations indicated and titrated down. CA125 was bound subsequently at a constant concentration. Detection was performed with a mouse anti-CA125 antibody and an MSD Sulfo tag labelled anti mouse FAB antibody.

[0029] FIG. 4 provides data on internalization of 125I-anti-mesothelin antibodies on CHO-A9 cells expressing mesothelin. Relative internalization of seven anti-mesothelin mabs, including the commercial positive control K1, in the absence (A), and in the presence (B) of the stabilizing second antibody. Representative data using MF-226 plus second antibody, showing relative amounts of dissociated, surface-bound and internalized antibody at 37° C. over time (C) is compared with that at the non-permissive temperature of 0° C. (D).

DETAILED DESCRIPTION OF THE INVENTION

[0030] The present invention is based on the discovery of novel antibodies that are specific to or have a high affinity for mesothelin and can deliver a therapeutic benefit to a subject. The antibodies of the invention, which may be human or humanized, can be used in many contexts, which are more fully described herein.

DEFINITIONS

[0031] A "human" antibody or functional human antibody fragment is hereby defined as one that is not chimeric (e.g., not "humanized") and not from (either in whole or in part) a non-human species. A human antibody or functional antibody fragment can be derived from a human or can be a synthetic human antibody. A "synthetic human antibody" is defined herein as an antibody having a sequence derived, in whole or in part, in silico from synthetic sequences that are based on the analysis of known human antibody sequences. In silico design of a human antibody sequence or fragment thereof can be achieved, for example, by analyzing a database of human antibody or antibody fragment sequences and devising a polypeptide sequence utilizing the data obtained therefrom. Another example of a human antibody or functional antibody fragment is one that is encoded by a nucleic acid isolated from a library of antibody sequences of human origin (i.e., such library being based on antibodies taken from a human natural source). Examples of human antibodies include HuCAL antibodies as described in Knappik et al., J. Mol. Biol. (2000) 296:57 and U.S. Pat. No. 6,300,064.

[0032] A "humanized antibody" or functional humanized antibody fragment is defined herein as one that is (i) derived from a non-human source (e.g., a transgenic mouse which bears a heterologous immune system), which antibody is based on a human germline sequence; or (ii) chimeric, wherein the variable domain is derived from a non-human origin and the constant domain is derived from a human origin or (iii) CDR-grafted, wherein the CDRs of the variable domain are from a non-human origin, while one or more frameworks of the variable domain are of human origin and the constant domain (if any) is of human origin.

[0033] As used herein, an antibody "binds specifically to," is "specific to/for" or "specifically recognizes" an antigen (here, mesothelin) if such antibody is able to discriminate between such antigen and one or more reference antigen(s), since binding specificity is not an absolute, but a relative property. In its most general form (and when no defined reference is mentioned), "specific binding" is referring to the ability of the antibody to discriminate between the antigen of interest and an unrelated antigen, as determined, for example, in accordance with one of the following methods. Such methods comprise, but are not limited to Western blots, ELISA-, RIA-, ECL-, IRMA-tests and peptide scans. For example, a standard ELISA assay can be carried out. The scoring may be carried out by standard color development (e.g. secondary antibody with horseradish peroxide and tetramethyl benzidine with hydrogenperoxide). The reaction in certain wells is scored by the optical density, for example, at 450 nm. Typical background (=negative reaction) may be 0.1 OD; typical positive reaction may be 1 OD. This means the difference positive/negative can be more than 10-fold. Typically, determination of binding specificity is performed by using not a single reference antigen, but a set of about three to five unrelated antigens, such as milk powder, BSA, transferrin or the like.

[0034] However, "specific binding" also may refer to the ability of an antibody to discriminate between the target antigen and one or more closely related antigen(s), which are used as reference points. Additionally, "specific binding" may relate to the ability of an antibody to discriminate between different parts of its target antigen, e.g. different domains or regions of mesothelin, such as epitopes in the N-terminal or in the C-terminal region of mesothelin, or between one or more key amino acid residues or stretches of amino acid residues of mesothelin.

[0035] Also, as used herein, an "immunoglobulin" (Ig) hereby is defined as a protein belonging to the class IgG, IgM, IgE, IgA, or IgD (or any subclass thereof), and includes all conventionally known antibodies and functional fragments thereof. A "functional fragment" or "antigen-binding antibody fragment" of an antibody/immunoglobulin hereby is defined as a fragment of an antibody/immunoglobulin (e.g., a variable region of an IgG) that retains the antigen-binding region. An "antigen-binding region" of an antibody typically is found in one or more hypervariable region(s) of an antibody, i.e., the CDR-1, -2, and/or -3 regions; however, the variable "framework" regions can also play an important role in antigen binding, such as by providing a scaffold for the CDRs. Preferably, the "antigen-binding region" comprises at least amino acid residues 4 to 103 of the variable light (VL) chain and 5 to 109 of the variable heavy (VH) chain, more preferably amino acid residues 3 to 107 of VL and 4 to 111 of VH, and particularly preferred are the complete VL and VH chains (amino acid positions 1 to 109 of VL and 1 to 113 of VH; numbering according to WO 97/08320). A preferred class of immunoglobulins for use in the present invention is IgG. "Functional fragments" of the invention include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules (scFv); and multispecific antibodies formed from antibody fragments (C. A. K Borrebaeck, editor (1995) Antibody Engineering (Breakthroughs in Molecular Biology), Oxford University Press; R. Kontermann & S. Duebel, editors (2001) Antibody Engineering (Springer Laboratory Manual), Springer Verlag). An antibody other than a "bispecific" or "bifunctional" antibody is understood to have each of its binding sites identical. The F(ab')2 or Fab may be engineered to minimize or completely remove the intermolecular disulphide interactions that occur between the Cm and CL domains.

[0036] An antibody of the invention may be derived from a recombinant antibody library that is based on amino acid sequences that have been designed in silico and encoded by nucleic acids that are synthetically created. In silico design of an antibody sequence is achieved, for example, by analyzing a database of human sequences and devising a polypeptide sequence utilizing the data obtained therefrom. Methods for designing and obtaining in silico-created sequences are described, for example, in Knappik et al., J. Mol. Biol. (2000) 296:57; Krebs et al., J. Immunol. Methods. (2001) 254:67; and U.S. Pat. No. 6,300,064 issued to Knappik et al., which hereby are incorporated by reference in their entirety.

[0037] As used herein, different `forms` of antigen, e.g. mesothelin, are hereby defined as different protein molecules resulting from different translational and posttranslational modifications, such as, but not limited to, differences in splicing of the primary mesothelin transcript, differences in glycosylation, and differences in posttranslational proteolytic cleavage.

[0038] As used herein, the term `invariant binding` of a particular antibody to mesothelin refers to its ability to bind to mesothelin on a broad range of mesothelin-expressing cancer cell lines which express different forms of mesothelin. For invariantly binding antibodies, EC50 values determined by FACS titration on two distinct cancer cell lines might differ no more than 10 fold, or, preferably, 5 fold, and most preferably between 1 and 3 fold.

[0039] As used herein, the term `epitope` includes any protein determinant capable of specific binding to an immunoglobulin or T-cell receptor. Epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics. Two antibodies are said to `bind the same epitope` if one antibody is shown to compete with the second antibody in a competitive binding assay, by any of the methods well known to those of skill in the art.

Antibodies of the Invention

[0040] The present invention relates to methods to inhibit growth of mesothelin-positive cancer cells and the progression of neoplastic disease by providing anti-mesothelin antibodies. Provided are human monoclonal antibodies, antigen-binding antibody fragments thereof, and variants of the antibodies and fragments, that specifically bind to the 40 kDa, C-terminal domain of the mesothelin precursor polypeptide (SEQ ID NO 370), which is named `mesothelin` herein.

[0041] The antibodies, antigen-binding antibody fragments, and variants of the antibodies and fragments of the invention are comprised of a light chain variable region and a heavy chain variable region. Variants of the antibodies or antigen-binding antibody fragments contemplated in the invention are molecules in which the binding activity of the antibody or antigen-binding antibody fragment for mesothelin is maintained.

[0042] Throughout this document, reference is made to the following representative antibodies of the invention: "MF-J", "MOR07265", "MOR06631", "MOR 06635", "MOR06669", "MOR07111", "MOR06640", "MOR06642", "MOR06643", "MF-226", "MOR06626", "MOR06638", "MF-A", "MOR06657", "MF-T", "MF1", "MF-5", "MF-8", "MF-24", "MF-25", "MF-27", "MF-73", "MF-78", "MF-84", "MF-101", "MF-230", "MF-236", "MF-252", "MF-257", "MF-423", "MF-427", "MF-428", MF-C'', "MF-I", "MF-L", "MF-M", "MF-P", "MF-Q", MF-S'', "MF-V", "MF-W", and "MF-Y". MF-J represents an antibody having a variable heavy region corresponding to SEQ ID NO: 284 (DNA)/SEQ ID NO: 198 (protein) and a variable light region corresponding to SEQ ID NO: 327 (DNA)/SEQ ID NO: 241 (protein). MOR 07265 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 285 (DNA)/SEQ ID NO: 199 (protein) and a variable light region corresponding to SEQ ID NO: 328 (DNA)/SEQ ID NO: 242 (protein). MOR 06631 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 286 (DNA)/SEQ ID NO: 200 (protein) and a variable light region corresponding to SEQ ID NO: 329 (DNA)/SEQ ID NO: 243 (protein). MOR 06669 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 287 (DNA)/SEQ ID NO: 201 (protein) and a variable light region corresponding to SEQ ID NO: 330 (DNA)/SEQ ID NO: 244 (protein). MOR 07111 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 288 (DNA)/SEQ ID NO: 202 (protein) and a variable light region corresponding to SEQ ID NO: 331 (DNA)/SEQ ID NO: 245 (protein). MOR 06640 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 289 (DNA)/SEQ ID NO: 203 (protein) and a variable light region corresponding to SEQ ID NO: 332 (DNA)/SEQ ID NO: 246 (protein). MOR 06642 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 290 (DNA)/SEQ ID NO: 204 (protein) and a variable light region corresponding to SEQ ID NO: 333 (DNA)/SEQ ID NO: 247 (protein). MOR 06643 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 291 (DNA)/SEQ ID NO: 205 (protein) and a variable light region corresponding to SEQ ID NO: 334 (DNA)/SEQ ID NO: 248 (protein). MF-226 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 292 (DNA)/SEQ ID NO: 206 (protein) and a variable light region corresponding to SEQ ID NO: 335 (DNA)/SEQ ID NO: 249 (protein). MOR 06626 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 293 (DNA)/SEQ ID NO: 207 (protein) and a variable light region corresponding to SEQ ID NO: 336 (DNA)/SEQ ID NO: 250 (protein). MOR 06635 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 294 (DNA)/SEQ ID NO: 208 (protein) and a variable light region corresponding to SEQ ID NO: 337 (DNA)/SEQ ID NO: 251 (protein). MOR 06638 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 295 (DNA)/SEQ ID NO: 209 (protein) and a variable light region corresponding to SEQ ID NO: 338 (DNA)/SEQ ID NO: 252 (protein). MF-A represents an antibody having a variable heavy region corresponding to SEQ ID NO: 296 (DNA)/SEQ ID NO: 210 (protein) and a variable light region corresponding to SEQ ID NO: 339 (DNA)/SEQ ID NO: 253 (protein). MOR 06657 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 297 (DNA)/SEQ ID NO: 211 (protein) and a variable light region corresponding to SEQ ID NO: 340 (DNA)/SEQ ID NO: 254 (protein). MF-T represents an antibody having a variable heavy region corresponding to SEQ ID NO: 298 (DNA)/SEQ ID NO: 212 (protein) and a variable light region corresponding to SEQ ID NO: 341 (DNA)/SEQ ID NO: 255 (protein). MF-L represents an antibody having a variable heavy region corresponding to SEQ ID NO: 299 (DNA)/SEQ ID NO: 213 (protein) and a variable light region corresponding to SEQ ID NO: 342 (DNA)/SEQ ID NO: 256 (protein). MF-1 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 300 (DNA)/SEQ ID NO: 214 (protein) and a variable light region corresponding to SEQ ID NO: 343 (DNA)/SEQ ID NO: 257 (protein). MF-5 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 301 (DNA)/SEQ ID NO: 215 (protein) and a variable light region corresponding to SEQ ID NO: 344 (DNA)/SEQ ID NO: 258 (protein). MF-8 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 302 (DNA)/SEQ ID NO: 216 (protein) and a variable light region corresponding to SEQ ID NO: 345 (DNA)/SEQ ID NO: 259 (protein). MF-24 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 303 (DNA)/SEQ ID NO: 217 (protein) and a variable light region corresponding to SEQ ID NO: 346 (DNA)/SEQ ID NO: 260 (protein). MF-25 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 304 (DNA)/SEQ ID NO: 218 (protein) and a variable light region corresponding to SEQ ID NO: 347 (DNA)/SEQ ID NO: 261 (protein). MF-27 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 305 (DNA)/SEQ ID NO: 219 (protein) and a variable light region corresponding to SEQ ID NO: 348 (DNA)/SEQ ID NO: 262 (protein). MF-73 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 306 (DNA)/SEQ ID NO: 220 (protein) and a variable light region corresponding to SEQ ID NO: 349 (DNA)/SEQ ID NO: 263 (protein). MF-78 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 307 (DNA)/SEQ ID NO: 221 (protein) and a variable light region corresponding to SEQ ID NO: 350 (DNA)/SEQ ID NO: 264 (protein). MF-84 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 308 (DNA)/SEQ ID NO: 222 (protein) and a variable light region corresponding to SEQ ID NO: 351 (DNA)/SEQ ID NO: 265 (protein). MF-101 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 309 (DNA)/SEQ ID NO: 223 (protein) and a variable light region corresponding to SEQ ID NO: 352 (DNA)/SEQ ID NO: 266 (protein). MF-230 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 310 (DNA)/SEQ ID NO: 224 (protein) and a variable light region corresponding to SEQ ID NO: 353 (DNA)/SEQ ID NO: 267 (protein). MF-236 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 311 (DNA)/SEQ ID NO: 225 (protein) and a variable light region corresponding to SEQ ID NO: 354 (DNA)/SEQ ID NO: 268 (protein). MF-252 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 312 (DNA)/SEQ ID NO: 226 (protein) and a variable light region corresponding to SEQ ID NO: 355 (DNA)/SEQ ID NO: 269 (protein). MF-275 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 313 (DNA)/SEQ ID NO: 227 (protein) and a variable light region corresponding to SEQ ID NO: 356 (DNA)/SEQ ID NO: 270 (protein). MF-423 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 314 (DNA)/SEQ ID NO: 228 (protein) and a variable light region corresponding to SEQ ID NO: 357 (DNA)/SEQ ID NO: 271 (protein). MF-427 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 315 (DNA)/SEQ ID NO: 229 (protein) and a variable light region corresponding to SEQ ID NO: 358 (DNA)/SEQ ID NO: 272 (protein). MF-428 represents an antibody having a variable heavy region corresponding to SEQ ID NO: 316 (DNA)/SEQ ID NO: 230 (protein) and a variable light region corresponding to SEQ ID NO: 359 (DNA)/SEQ ID NO: 273 (protein). MF-C represents an antibody having a variable heavy region corresponding to SEQ ID NO: 317 (DNA)/SEQ ID NO: 231 (protein) and a variable light region corresponding to SEQ ID NO: 360 (DNA)/SEQ ID NO: 274 (protein). MF-I represents an antibody having a variable heavy region corresponding to SEQ ID NO: 318 (DNA)/SEQ ID NO: 232 (protein) and a variable light region corresponding to SEQ ID NO: 361 (DNA)/SEQ ID NO: 275 (protein). MF-M represents an antibody having a variable heavy region corresponding to SEQ ID NO: 319 (DNA)/SEQ ID NO: 233 (protein) and a variable light region corresponding to SEQ ID NO: 362 (DNA)/SEQ ID NO: 276 (protein). MF-P represents an antibody having a variable heavy region corresponding to SEQ ID NO: 320 (DNA)/SEQ ID NO: 234 (protein) and a variable light region corresponding to SEQ ID NO: 363 (DNA)/SEQ ID NO: 277 (protein). MF-Q represents an antibody having a variable heavy region corresponding to SEQ ID NO: 321 (DNA)/SEQ ID NO: 235 (protein) and a variable light region corresponding to SEQ ID NO: 364 (DNA)/SEQ ID NO: 278 (protein). MF-S represents an antibody having a variable heavy region corresponding to SEQ ID NO: 322 (DNA)/SEQ ID NO: 236 (protein) and a variable light region corresponding to SEQ ID NO: 365 (DNA)/SEQ ID NO: 279 (protein). MF-U represents an antibody having a variable heavy region corresponding to SEQ ID NO: 323 (DNA)/SEQ ID NO: 237 (protein) and a variable light region corresponding to SEQ ID NO: 366 (DNA)/SEQ ID NO: 280 (protein). MF-V represents an antibody having a variable heavy region corresponding to SEQ ID NO: 324 (DNA)/SEQ ID NO: 238 (protein) and a variable light region corresponding to SEQ ID NO: 367 (DNA)/SEQ ID NO: 281 (protein). MF-W represents an antibody having a variable heavy region corresponding to SEQ ID NO: 325 (DNA)/SEQ ID NO: 239 (protein) and a variable light region corresponding to SEQ ID NO: 368 (DNA)/SEQ ID NO: 282 (protein). MF-Y represents an antibody having a variable heavy region corresponding to SEQ ID NO: 326 (DNA)/SEQ ID NO: 240 (protein) and a variable light region corresponding to SEQ ID NO: 369 (DNA)/SEQ ID NO: 283 (protein).

[0043] In one aspect, the invention provides antibodies which bind to epitopes of mesothelin, whose amino acid sequence is depicted by SEQ ID NO: 370, that are distinct from the mesothelin epitope recognized by Mab K1.

[0044] In other aspects the invention provides antibodies which bind to one or more amino acids of the epitopes of antibodies MF-J or MF-T. In certain aspects said antibodies bind to at least to two, at least three, at least four, at least five or at least six amino acids of the epitopes of antibodies MF-J or MF-T. In certain aspects the antibodies of the present invention bind to one or more amino acids of the epitope recognized by the antibody MF-J. In alternative aspects the antibodies of the present invention bind to one or more amino acids of the epitope recognized by the antibody MF-T.

[0045] In another aspect, the invention provides antibodies having an antigen-binding region that can bind specifically to or has a high affinity for one or more regions of mesothelin, whose amino acid sequence is depicted by SEQ ID NO: 370. An antibody is said to have a "high affinity" for an antigen if the affinity measurement is at least 100 nM (monovalent affinity of Fab fragment). An inventive antibody or antigen-binding region preferably can bind to mesothelin with an affinity of less than about 100 nM, more preferably less than about 60 nM, and still more preferably less than about 30 nM. Further preferred are antibodies that bind to mesothelin with an affinity of less than about 10 nM, and more preferably less than about 3 nM. For instance, the affinity of an antibody of the invention against mesothelin may be about 10.0 nM or 0.19 nM (monovalent affinity of Fab fragment).

[0046] Table 1 provides a summary of dissociation constants and dissociation rates of representative antibodies of the invention, as determined by surface plasmon resonance (Biacore) on directly immobilized mesothelin.

TABLE-US-00001 TABLE 1 Monovalent dissociation constants and dissociation rates to mesothelin determined for anti-mesothelin Fabs by surface plasmon resonance Antibody K_D [M] kd [1/s] MF-A 1.9 × 10^-8 7.9 × 10^-2 MOR06657 9.5 × 10^-10 5.5 × 10^-3 MF-J 9.2 × 10^-9 2.9 × 10^-3 MOR06631 9 × 10^-11 1.4 × 10^-5 MOR06669 2.4 × 10^-10 8.1 × 10^-5 MOR06643 3.6 × 10^-10 2.8 × 10^-4 MF-226 5.8 × 10^-8 3.8 × 10^-2 MOR06626 6.7 × 10^-10 1.2 × 10^-3 MOR06638 1.6 × 10^-8 6.3 × 10^-3

[0047] The IgG1 format was used for the cell-based affinity determination, determined by fluorescence-activated cell sorting (FACS) combined with Scatchard analysis, and live cell enzyme-linked immunosorbed assay (ELISA). Table 2 denotes the binding strength of representative IgG antibodies on mesothelin-expressing CHO-A9 cells.

TABLE-US-00002 TABLE 2 Cell-based binding potency of anti-mesothelin antibodies as determined by cell ELISA and FACS on mesothelin-expressing CHO-A9 cells EC50 Antibody (IgG) FACS [nM] Cell ELISA [nM] MF-A 0.05 0.8 MF-J 0.11 3.9 MF-L 0.07 0.8 MF-T 0.27 1.5 MF-226 0.15 0.4

[0048] Antibody Generation

[0049] A synthetic antibody phage display library (Knappik, A., et al., J. Mol. Biol. (2000) 296(1): 57) was used to isolate high affinity, mesothelin-specific, human monoclonal antibodies, by a combination of whole cell and protein pannings and through the development of specific tools. These tools and methods include a mesothelin-expressing recombinant cell-line and the development of panning procedures and screening assays capable of identifying antibodies that preferentially bind to mesothelin displayed on the cell surface and that are crossreactive to mesothelin from other species.

[0050] Antibodies to the mesothelial cancer cell-surface marker, mesothelin, were discovered by a combination of three non-conventional approaches in phage-display technology (PDT). First, a recombinant cell line expressing the membrane-bound, 40 kDa domain of mesothelin was constructed by stable transfection of CHO-K1 cells with a plasmid encoding the GPI-anchored C-terminal part of the protein (SEQ ID 371), to give the CHO-A9 cell line. Second, dual-alternating cell-surface selections were performed with the latter recombinant cell line and the squamous cancer cell line NCI-H226. Pre-adsorption with CHO-K1 cells was included to avoid the selection of Fab fragments binding to epitopes of the parental cells. Additional selections were performed with recombinant, soluble purified human mesothelin (unique source of "MF-24", "MF-25", and "MF-27"), with recombinant, murine mesothelin, with purified deglycosylated mesothelin (unique source of "MF-5" and "MF-8"), and with biotinylated mesothelin in soluble phase. Third, screening methods were developed which allowed for successive screening of the phage outputs obtained in panning on whole NCI-H226 cells as well as CHO-A9 cells. The combination of these specific methods allowed the isolation of the unique antibodies "MF-J", "MF-226", "MF-A", "MF-T", "MF-1", "MF-5", "MF-8", "MF-24", "MF-25", "MF-27", "MF-73", "MF-78", "MF-84", "MF-101", "MF-230", "MF-236", "MF-252", "MF-275", "MF-423", "MF-427", "MF-428", MF-C'', "MF-I", "MF-L", "MF-M", "MF-P", "MF-Q", MF-S'', "MF-U", "MF-V", "MF-W", and "MF-Y". These unique antibodies were further characterized by their binding affinity in two cell based ELISA's, by BIAcore binding to soluble mesothelin, by their ability to recognize different epitopes on soluble mesothelin, and by their ability to cross react with murine mesothelin assessed by FACS and immunoblotting, and their ability to be internalized in three different cell based assays. Two of the internalization assays quantitatively measured the internalization of radiolabelled anti-mesothelin antibodies either in the absence of presence of a secondary antibody to human IgG. This data was used to select four antibodies for further affinity maturation.

[0051] In order to obtain antibodies with robust invariant binding to different forms of mesothelin displayed on different cancer cell lines, to increase species cross-reactivity, and to further increase affinity and decrease dissociation rates, a strategy for affinity maturation was designed. Affinity maturation was performed on antibodies `MF-J`, `MF-226`, `MF-L` and `MF-A`. Affinity maturation included generation of new antibody repertoires by the exchange of H-CDR2, L-CDR3, or a combination of both H-CDR2 and L-CDR3 regions of the parental antibodies. Alternating selections were performed with the two mesothelin-expressing cancer cell lines NCI-H226 and OVCAR-3, as well as recombinant purified and biotinylated human and murine meothelin in solution using magnetic beads. Increasing stringency was obtained by gradual reduction of antigen and extension of the washing procedure.

[0052] Screening was performed by first ranking the hits by decreasing affinity, as determined on antigen-coated beads in solution, by measuring an electrochemiluminescent signal in a M-384 Workstation (BioVeris). Subsequently, a resulting selection of high-affinity binders was submitted to solution-equilibrium titration (SET) screening (Haenel, C., et al., Anal. Biochem. (2005) 339(1): 182). The best binders were further screened by analysis of cross-reactivity to murine mesothelin, as well as for binding to mesothelin on NCI-H226 cells by FACS. The combination of these specific methods allowed the isolation of the unique antibodies `MOR07265`, `MOR06631`, `MOR 06635`, `MOR06669`, `MOR07111`, `MOR06640`, `MOR06642`, `MOR06643`, `MOR06626`, `MOR06638` and `MOR06657`.

Peptide Variants

[0053] Antibodies of the invention are not limited to the specific peptide sequences provided herein. Rather, the invention also embodies variants of these polypeptides. With reference to the instant disclosure and conventionally available technologies and references, the skilled worker will be able to prepare, test and utilize functional variants of the antibodies disclosed herein, while appreciating that variants having the ability to bind to mesothelin fall within the scope of the present invention.

[0054] A variant can include, for example, an antibody that has at least one altered complementary determining region (CDR) (hyper-variable) and/or framework (FR) (variable) domain/position, vis-a-vis a peptide sequence disclosed herein. To better illustrate this concept, a brief description of antibody structure follows.

[0055] An antibody is composed of two peptide chains, each containing one (light chain) or three (heavy chain) constant domains and a variable region (VL, VH), the latter of which is in each case made up of four FR regions and three interspaced CDRs. The antigen-binding site is formed by one or more CDRs, yet the FR regions provide the structural framework for the CDRs and, hence, play an important role in antigen binding. By altering one or more amino acid residues in a CDR or FR region, the skilled worker routinely can generate mutated or diversified antibody sequences, which can be screened against the antigen, for new or improved properties, for example.

[0056] Tables 3 (VH) and 4 (VL) delineate the CDR and FR regions for certain antibodies of the invention and compare amino acids at a given position to each other and to corresponding consensus or "master gene" sequences (as described in U.S. Pat. No. 6,300,064):

TABLE-US-00003 TABLE 3 VH Sequences ##STR00001## ##STR00002## ##STR00003##

TABLE-US-00004 TABLE 4 VL Sequences ##STR00004## ##STR00005## ##STR00006##

[0057] In certain aspects the present invention provides antibodies

[0058] wherein the HCDR1 region is selected from sequence ID's [all respective SEQ IDs of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30.

[0059] wherein the HCDR2 region is selected from sequence ID's 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65 or 66.

[0060] wherein the HCDR3 region is selected from sequence ID's 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97 or 98.

[0061] wherein the LCDR1 region is selected from sequence ID's 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 102 or 128.

[0062] wherein the LCDR2 region is selected from sequence ID's 129, 130, 131 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159 or 155.

[0063] wherein the LCDR3 region is selected from sequence ID's 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196 or 197. or combinations of these CDR regions.

[0064] Preferred aspects are antibodies: in which the CDR sequences are selected from the MF-J series as shown in table 7 or other combinations of the CDR regions shown in table 7.

[0065] In certain aspects the present invention provides antibodies

[0066] wherein the VH is selected from sequence ID 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239 or 240,

[0067] wherein the VL is selected from sequence ID 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282 or 283.

[0068] As above, preferred aspects for MF-J series as shown in table 7 or other combinations of the VH and VL regions shown in table 7.

[0069] The skilled worker can use the data in Tables 3, 4 and 7 to design peptide variants that are within the scope of the present invention. It is preferred that variants are constructed by changing amino acids within one or more CDR regions; a variant might also have one or more altered framework regions. With reference to a comparison of the novel antibodies to each other, candidate residues that can be changed include e.g. residues 3 or 45 of the variable light and e.g. residues 16 or 43 of the variable heavy chains of MF-226 and MF-T, since these are positions of variance vis-a-vis each other. Alterations also may be made in the framework regions. For example, a peptide FR domain might be altered where there is a deviation in a residue compared to a germline sequence.

[0070] With reference to a comparison of the novel antibodies to the corresponding consensus or "master gene" sequence, which are listed in Knappik et al., 2000, candidate residues that can be changed include e.g. residues 29 or 52 of the variable light chain of MF-T compared to VLX2 and e.g. residues 43 or 57 of the variable heavy chain of MF-A compared to VH1A (Knappik, A., et al., J. Mol. Biol. (2000) 296(1): 57). Alternatively, the skilled worker could make the same analysis by comparing the amino acid sequences disclosed herein to known sequences of the same class of such antibodies, using, for example, the procedure described by Knappik, A., et al. (2000) and U.S. Pat. No. 6,300,064 issued to Knappik et al.

[0071] Furthermore, variants may be obtained by using one antibody as starting point for optimization by diversifying one or more amino acid residues in the antibody, preferably amino acid residues in one or more CDRs, and by screening the resulting collection of antibody variants for variants with improved properties. Particularly preferred is diversification of one or more amino acid residues in CDR-3 of VL, CDR-3 of VH, CDR-1 of VL and/or CDR-2 of VH. Diversification can be done by synthesizing a collection of DNA molecules using trinucleotide mutagenesis (TRIM) technology (Virnekas, B., Ge, L., Pluckthun, A., Schneider, K. C., Wellnhofer, G., and Moroney S. E. (1994) Trinucleotide phosphoramidites: ideal reagents for the synthesis of mixed oligonucleotides for random mutagenesis. Nucl. Acids Res. 22, 5600.).

Conservative Amino Acid Variants

[0072] Polypeptide variants may be made that conserve the overall molecular structure of an antibody peptide sequence described herein. Given the properties of the individual amino acids, some rational substitutions will be recognized by the skilled worker. Amino acid substitutions, i.e., "conservative substitutions," may be made, for instance, on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues involved.

[0073] For example, (a) nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; (b) polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; (c) positively charged (basic) amino acids include arginine, lysine, and histidine; and (d) negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Substitutions typically may be made within groups (a)-(d). In addition, glycine and proline may be substituted for one another based on their ability to disrupt α-helices. Similarly, certain amino acids, such as alanine, cysteine, leucine, methionine, glutamic acid, glutamine, histidine and lysine are more commonly found in α-helices, while valine, isoleucine, phenylalanine, tyrosine, tryptophan and threonine are more commonly found in β-pleated sheets. Glycine, serine, aspartic acid, asparagine, and proline are commonly found in turns. Some preferred substitutions may be made among the following groups: (i) S and T; (ii) P and G; and (iii) A, V, L and I. Given the known genetic code, and recombinant and synthetic DNA techniques, the skilled scientist readily can construct DNAs encoding the conservative amino acid variants. In one particular example, amino acid position 3 in SEQ ID NOS: 199-205, 207-211 or 213-240 can be changed from a Q to an E.

[0074] As used herein, "sequence identity" between two polypeptide sequences, indicates the percentage of amino acids that are identical between the sequences. "Sequence homology" indicates the percentage of amino acids that either are identical or that represent conservative amino acid substitutions. Preferred polypeptide sequences of the invention have a sequence identity in the CDR regions of at least 60%, more preferably, at least 70% or 80%, still more preferably at least 90% and most preferably at least 95%. Preferred antibodies also have a sequence homology in the CDR regions of at least 80%, more preferably 90% and most preferably 95%.

DNA Molecules of the Invention

[0075] The present invention also relates to the DNA molecules that encode an antibody of the invention. These sequences include, but are not limited to, those DNA molecules set forth in SEQ IDs 284-369.

[0076] DNA molecules of the invention are not limited to the sequences disclosed herein, but also include variants thereof. DNA variants within the invention may be described by reference to their physical properties in hybridization. The skilled worker will recognize that DNA can be used to identify its complement and, since DNA is double stranded, its equivalent or homolog, using nucleic acid hybridization techniques. It also will be recognized that hybridization can occur with less than 100% complementarity. However, given appropriate choice of conditions, hybridization techniques can be used to differentiate among DNA sequences based on their structural relatedness to a particular probe. For guidance regarding such conditions see, Sambrook et al., 1989 (Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A laboratory manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, USA) and Ausubel et al., 1995 (Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Sedman, J. G., Smith, J. A., & Struhl, K. eds. (1995). Current Protocols in Molecular Biology. New York: John Wiley and Sons).

[0077] Structural similarity between two polynucleotide sequences can be expressed as a function of "stringency" of the conditions under which the two sequences will hybridize with one another. As used herein, the term "stringency" refers to the extent that the conditions disfavor hybridization. Stringent conditions strongly disfavor hybridization, and only the most structurally related molecules will hybridize to one another under such conditions. Conversely, non-stringent conditions favor hybridization of molecules displaying a lesser degree of structural relatedness. Hybridization stringency, therefore, directly correlates with the structural relationships of two nucleic acid sequences. The following relationships are useful in correlating hybridization and relatedness (where T_m is the melting temperature of a nucleic acid duplex):

[0078] a. T_m=69.3+0.41(G+C)%

[0079] b. The T_m of a duplex DNA decreases by 1° C. with every increase of 1% in the number of mismatched base pairs.

[0080] c. (T_m).sub.μ2-(T_m).sub.μ1=18.5 log₁₀μ2/μl

[0081] where μ1 and μ2 are the ionic strengths of two solutions.

[0082] Hybridization stringency is a function of many factors, including overall DNA concentration, ionic strength, temperature, probe size and the presence of agents which disrupt hydrogen bonding. Factors promoting hybridization include high DNA concentrations, high ionic strengths, low temperatures, longer probe size and the absence of agents that disrupt hydrogen bonding. Hybridization typically is performed in two phases: the "binding" phase and the "washing" phase.

[0083] First, in the binding phase, the probe is bound to the target under conditions favoring hybridization. Stringency is usually controlled at this stage by altering the temperature. For high stringency, the temperature is usually between 65° C. and 70° C., unless short (<20 nt) oligonucleotide probes are used. A representative hybridization solution comprises 6×SSC, 0.5% SDS, 5×Denhardt's solution and 100 μg of nonspecific carrier DNA. See Ausubel et al., section 2.9, supplement 27 (1994). Of course, many different, yet functionally equivalent, buffer conditions are known. Where the degree of relatedness is lower, a lower temperature may be chosen. Low stringency binding temperatures are between about 25° C. and 40° C. Medium stringency is between at least about 40° C. to less than about 65° C. High stringency is at least about 65° C.

[0084] Second, the excess probe is removed by washing. It is at this phase that more stringent conditions usually are applied. Hence, it is this "washing" stage that is most important in determining relatedness via hybridization. Washing solutions typically contain lower salt concentrations. One exemplary medium stringency solution contains 2×SSC and 0.1% SDS. A high stringency wash solution contains the equivalent (in ionic strength) of less than about 0.2×SSC, with a preferred stringent solution containing about 0.1×SSC. The temperatures associated with various stringencies are the same as discussed above for "binding." The washing solution also typically is replaced a number of times during washing. For example, typical high stringency washing conditions comprise washing twice for 30 minutes at 55° C. and three times for 15 minutes at 60° C.

[0085] Accordingly, the present invention includes nucleic acid molecules that hybridize to the molecules of set forth in SEQ ID 284-369 under high stringency binding and washing conditions, where such nucleic molecules encode an antibody or functional fragment thereof having properties as described herein. Preferred molecules (from an mRNA perspective) are those that have at least 75% or 80% (preferably at least 85%, more preferably at least 90% and most preferably at least 95%) homology or sequence identity with one of the DNA molecules described herein. In one particular example of a variant of the invention, nucleic acid position 7 in SEQ ID NOS: 285-291, 293-297, or 299-326 can be substituted from a C to a G, thereby changing the codon from CAA to GAA.

Functionally Equivalent Variants

[0086] Yet another class of DNA variants within the scope of the invention may be described with reference to the product they encode. These functionally equivalent genes are characterized by the fact that they encode the same peptide sequences found in SEQ ID 284-369 due to the degeneracy of the genetic code.

[0087] It is recognized that variants of DNA molecules provided herein can be constructed in several different ways. For example, they may be constructed as completely synthetic DNAs. Methods of efficiently synthesizing oligonucleotides in the range of 20 to about 150 nucleotides are widely available. See Ausubel et al., section 2.11, Supplement 21 (1993). Overlapping oligonucleotides may be synthesized and assembled in a fashion first reported by Khorana et al., J. Mol. Biol. 72:209-217 (1971); see also Ausubel et al., supra, Section 8.2. Synthetic DNAs preferably are designed with convenient restriction sites engineered at the 5' and 3' ends of the gene to facilitate cloning into an appropriate vector.

[0088] As indicated, a method of generating variants is to start with one of the DNAs disclosed herein and then to conduct site-directed mutagenesis. See Ausubel et al., supra, chapter 8, Supplement 37 (1997). In a typical method, a target DNA is cloned into a single-stranded DNA bacteriophage vehicle. Single-stranded DNA is isolated and hybridized with an oligonucleotide containing the desired nucleotide alteration(s). The complementary strand is synthesized and the double stranded phage is introduced into a host. Some of the resulting progeny will contain the desired mutant, which can be confirmed using DNA sequencing. In addition, various methods are available that increase the probability that the progeny phage will be the desired mutant. These methods are well known to those in the field and kits are commercially available for generating such mutants.

Recombinant DNA Constructs and Expression

[0089] The present invention further provides recombinant DNA constructs comprising one or more of the nucleotide sequences of the present invention. The recombinant constructs of the present invention are used in connection with a vector, such as a plasmid, phagemid, phage or viral vector, into which a DNA molecule encoding an antibody of the invention is inserted.

[0090] The encoded gene may be produced by techniques described in Sambrook et al., 1989, and Ausubel et al., 1989. Alternatively, the DNA sequences may be chemically synthesized using, for example, synthesizers. See, for example, the techniques described in OLIGONUCLEOTIDE SYNTHESIS (1984, Gait, ed., IRL Press, Oxford), which is incorporated by reference herein in its entirety. Recombinant constructs of the invention are comprised with expression vectors that are capable of expressing the RNA and/or protein products of the encoded DNA(s). The vector may further comprise regulatory sequences, including a promoter operably linked to the open reading frame (ORF). The vector may further comprise a selectable marker sequence. Specific initiation and bacterial secretory signals also may be required for efficient translation of inserted target gene coding sequences.

[0091] The present invention further provides host cells containing at least one of the DNAs of the present invention. The host cell can be virtually any cell for which expression vectors are available. It may be, for example, a higher eukaryotic host cell, such as a mammalian cell, a lower eukaryotic host cell, such as a yeast cell, and may be a prokaryotic cell, such as a bacterial cell. Introduction of the recombinant construct into the host cell can be effected by calcium phosphate transfection, DEAE, dextran mediated transfection, electroporation or phage infection.

Bacterial Expression

[0092] Useful expression vectors for bacterial use are constructed by inserting a structural DNA sequence encoding a desired protein together with suitable translation initiation and termination signals in operable reading phase with a functional promoter. The vector will comprise one or more phenotypic selectable markers and an origin of replication to ensure maintenance of the vector and, if desirable, to provide amplification within the host. Suitable prokaryotic hosts for transformation include E. coli, Bacillus subtilis, Salmonella typhimurium and various species within the genera Pseudomonas, Streptomyces, and Staphylococcus.

[0093] Bacterial vectors may be, for example, bacteriophage-, plasmid- or phagemid-based. These vectors can contain a selectable marker and bacterial origin of replication derived from commercially available plasmids typically containing elements of the well known cloning vector pBR322 (ATCC 37017). Following transformation of a suitable host strain and growth of the host strain to an appropriate cell density, the selected promoter is de-repressed/induced by appropriate means (e.g., temperature shift or chemical induction) and cells are cultured for an additional period. Cells are typically harvested by centrifugation, disrupted by physical or chemical means, and the resulting crude extract retained for further purification.

[0094] In bacterial systems, a number of expression vectors may be advantageously selected depending upon the use intended for the protein being expressed. For example, when a large quantity of such a protein is to be produced, for the generation of antibodies or to screen peptide libraries, for example, vectors which direct the expression of high levels of fusion protein products that are readily purified may be desirable.

Therapeutic Methods

[0095] Therapeutic methods involve administering to a subject in need of treatment a therapeutically effective amount of an antibody contemplated by the invention. A "therapeutically effective" amount hereby is defined as the amount of an antibody that is of sufficient quantity to deplete mesothelin-positive cells in a treated area of a subject--either as a single dose or according to a multiple dose regimen, alone or in combination with other agents, which leads to the alleviation of an adverse condition, yet which amount is toxicologically tolerable. The subject may be a human or non-human animal (e.g., rabbit, rat, mouse, monkey or other lower-order primate).

[0096] An antibody of the invention might be co-administered with known medicaments, and in some instances the antibody might itself be modified. For example, an antibody could be conjugated to an immunotoxin or radioisotope to potentially further increase efficacy.

[0097] The inventive antibodies can be used as a therapeutic or a diagnostic tool in a variety of situations where mesothelin undesirably expressed or found. Disorders and conditions particularly suitable for treatment with an antibody of the inventions are pancreatic cancer, ovarian cancer, mesothelioma and lung cancer.

[0098] To treat any of the foregoing disorders, pharmaceutical compositions for use in accordance with the present invention may be formulated in a conventional manner using one or more physiologically acceptable carriers or excipients. An antibody of the invention can be administered by any suitable means, which can vary, depending on the type of disorder being treated. Possible administration routes include parenteral (e.g., intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous), intrapulmonary and intranasal, and, if desired for local immunosuppressive treatment, intralesional administration. In addition, an antibody of the invention might be administered by pulse infusion, with, e.g., declining doses of the antibody. Preferably, the dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic. The amount to be administered will depend on a variety of factors such as the clinical symptoms, weight of the individual, whether other drugs are administered. The skilled artisan will recognize that the route of administration will vary depending on the disorder or condition to be treated.

[0099] Determining a therapeutically effective amount of the novel polypeptide, according to this invention, largely will depend on particular patient characteristics, route of administration, and the nature of the disorder being treated. General guidance can be found, for example, in the publications of the International Conference on Harmonisation and in REMINGTON'S PHARMACEUTICAL SCIENCES, chapters 27 and 28, pp. 484-528 (18th ed., Alfonso R. Gennaro, Ed., Easton, Pa.: Mack Pub. Co., 1990). More specifically, determining a therapeutically effective amount will depend on such factors as toxicity and efficacy of the medicament. Toxicity may be determined using methods well known in the art and found in the foregoing references. Efficacy may be determined utilizing the same guidance in conjunction with the methods described below in the Examples.

Diagnostic Methods

[0100] Mesothelin antibodies can be used for detecting the presence of mesothelin-expressing tumors. The presence of mesothelin-containing cells within various biological samples, including serum, prostate and other tissue biopsy specimens, may be detected with mesothelin antibodies. In addition, mesothelin antibodies may be used in various imaging methodologies such as immunoscintigraphy with a ⁹⁹mTc (or other isotope) conjugated antibody. For example, an imaging protocol similar to the one recently described using a ¹¹lIn conjugated anti-PSMA antibody may be used to detect pancreaetic or ovarian carcinomas (Sodee et al., Clin. Nuc. Med. 21: 759-766, 1997). Another method of detection that can be used is positron emitting tomography (see Herzog et al., J. Nucl. Med. 34:2222-2226, 1993).

Pharmaceutical Compositions and Administration

[0101] The present invention also relates to pharmaceutical compositions which may comprise mesothelin antibodies, alone or in combination with at least one other agent, such as stabilizing compound, which may be administered in any sterile, biocompatible pharmaceutical carrier, including, but not limited to, saline, buffered saline, dextrose, and water. Any of these molecules can be administered to a patient alone, or in combination with other agents, drugs or hormones, in pharmaceutical compositions where it is mixed with excipient(s) or pharmaceutically acceptable carriers. In one embodiment of the present invention, the pharmaceutically acceptable carrier is pharmaceutically inert.

[0102] The present invention also relates to the administration of pharmaceutical compositions. Such administration is accomplished orally or parenterally. Methods of parenteral delivery include topical, intra-arterial (directly to the tumor), intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, or intranasal administration. In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers comprising excipients and auxilliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Further details on techniques for formulation and administration may be found in the latest edition of Remington's Pharmaceutical Sciences (Ed. Maack Publishing Co, Easton, Pa.).

[0103] Pharmaceutical compositions for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration. Such carriers enable the pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for ingestion by the patient.

[0104] Pharmaceutical preparations for oral use can be obtained through combination of active compounds with solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxilliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are carbohydrate or protein fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato, or other plants; cellulose such as methyl, cellulose, hydroxypropylmethylcellulose, or sodium carboxymethylcellulose; and gums including arabic and tragacanth; and proteins such as gelatin and collagen. If desired, disintegrating or solubilizing agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate.

[0105] Dragee cores are provided with suitable coatings such as concentrated sugar solutions, which may also contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for product identification or to characterize the quantity of active compound, ie. dosage.

[0106] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a coating such as glycerol or sorbitol. Push-fit capsules can contain active ingredients mixed with a filler or binders such as lactose or starches, lubricants such as talc or magnesium stearate, and optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycol with or without stabilizers.

[0107] Pharmaceutical formulations for parenteral administration include aqueous solutions of active compounds. For injection, the pharmaceutical compositions of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiologically buffered saline. Aqueous injection suspensions may contain substances that increase viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[0108] For topical or nasal administration, penetrants appropriate to the particular barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

Kits

[0109] The invention further relates to pharmaceutical packs and kits comprising one or more containers filled with one or more of the ingredients of the aforementioned compositions of the invention. Associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, reflecting approval by the agency of the manufacture, use or sale of the product for human administration.

[0110] In another embodiment, the kits may contain DNA sequences encoding the antibodies of the invention. Preferably the DNA sequences encoding these antibodies are provided in a plasmid suitable for transfection into and expression by a host cell. The plasmid may contain a promoter (often an inducible promoter) to regulate expression of the DNA in the host cell. The plasmid may also contain appropriate restriction sites to facilitate the insertion of other DNA sequences into the plasmid to produce various antibodies. The plasmids may also contain numerous other elements to facilitate cloning and expression of the encoded proteins. Such elements are well known to those of skill in the art and include, for example, selectable markers, initiation codons, termination codons, and the like.

Manufacture and Storage.

[0111] The pharmaceutical compositions of the present invention may be manufactured in a manner that is known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.

[0112] The pharmaceutical composition may be provided as a salt and can be formed with may acids, including by not limited to hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend to be more soluble in aqueous or other protonic solvents that are the corresponding free base forms. In other cases, the preferred preparation may be a lyophilized powder in 1 mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol at a pH range of 4.5 to 5.5 that is combined with buffer prior to use.

[0113] After pharmaceutical compositions comprising a compound of the invention formulated in an acceptable carrier have been prepared, they can be placed in an appropriate container and labeled for treatment of an indicated condition. For administration of mesothelin antibodies, such labeling would include amount, frequency and method of administration.

Therapeutically Effective Dose.

[0114] Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose, i.e. treatment of a particular disease state characterized by mesothelin expression. The determination of an effective dose is well within the capability of those skilled in the art.

[0115] For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays, e.g., neoplastic cells, or in animal models, usually mice, rabbits, dogs, or pigs. The animal model is also used to achieve a desirable concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans.

[0116] A therapeutically effective dose refers to that amount of protein or its antibodies, antagonists, or inhibitors that ameliorate the symptoms or condition. Therapeutic efficacy and toxicity of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., ED₅₀ (the dose therapeutically effective in 50% of the population) and LD₅₀ (the dose lethal to 50% of the population). The dose ratio between therapeutic and toxic effects is the therapeutic index, and it can be expressed as the ratio, ED₅₀/LD₅₀. Pharmaceutical compositions that exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used in formulating a range of dosage for human use. The dosage of such compounds lies preferably within a range of circulating concentrations what include the ED₅₀ with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, sensitivity of the patient, and the route of administration.

[0117] The exact dosage is chosen by the individual physician in view of the patient to be treated. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Additional factors that may be taken into account include the severity of the disease state, eg, tumor size and location; age, weight and gender of the patient; diet, time and frequency of administration, drug combination(s), reaction sensitivities, and tolerance/response to therapy. Long acting pharmaceutical compositions might be administered every 3 to 4 days, every week, or once every two weeks depending on half-life and clearance rate of the particular formulation.

[0118] Normal dosage amounts may vary from 0.1 to 100,000 micrograms, up to a total dose of about 1 g, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature. See U.S. Pat. Nos. 4,657,760; 5,206,344; or 5,225,212. Those skilled in the art will employ different formulations for polynucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc. Preferred specific activities for for a radiolabeled antibody may range from 0.1 to 10 mCi/mg of protein (Riva et al., Clin. Cancer Res. 5:3275s-3280s, 1999; Wong et al., Clin. Cancer Res. 6:3855-3863, 2000; Wagner et al., J. Nuclear Med. 43:267-272, 2002).

[0119] The present invention is further described by the following examples. The examples are provided solely to illustrate the invention by reference to specific embodiments. These exemplifications, while illustrating certain specific aspects of the invention, do not portray the limitations or circumscribe the scope of the disclosed invention.

[0120] All examples were carried out using standard techniques, which are well known and routine to those of skill in the art, except where otherwise described in detail. Routine molecular biology techniques of the following examples can be carried out as described in standard laboratory manuals, such as Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed.; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.

EXAMPLES

Example 1

Antibody Generation from HuCAL Libraries

[0121] For the generation of therapeutic antibodies against mesothelin, selections with the MorphoSys HuCAL GOLD phage display library were carried out. HuCAL GOLD® is a Fab library based on the HuCAL® concept (Knappik, A., et al., J. Mol. Biol. (2000) 296(1): 57; Krebs, B., et al., J. Immunol. Methods. (2001) 254(1-2): 67), in which all six CDRs are diversified, and which employs the CysDisplay® technology for linking Fab fragments to the phage surface (Lohning, 2001; WO 01/05950).

A. Phagemid Rescue, Phage Amplification and Purification

[0122] HuCAL GOLD® phagemid library was amplified in 2×TY medium containing 34 μg/ml chloramphenicol and 1% glucose (2×TY-CG). After helper phage infection (VCSM13) at an OD600 of 0.5 (30 min at 37° C. without shaking; 30 min at 37° C. shaking at 250 rpm), cells were spun down (4120 g; 5 min; 4° C.), resuspended in 2×TY/34 μg/ml chloramphenicol/50 μg/ml kanamycin and grown overnight at 22° C. Phages were PEG-precipitated from the supernatant, resuspended in PBS/20% glycerol and stored at -80° C. Phage amplification between two panning rounds was conducted as follows: mid-log phase TG1 cells were infected with eluted phages and plated onto LB-agar supplemented with 1% of glucose and 34 μg/ml of chloramphenicol (LB-CG). After overnight incubation at 30° C., colonies were scraped off, adjusted to an OD600 of 0.5 and helper phage added as described above.

B. Pannings with HuCAL GOLD®

[0123] For the selections HuCAL GOLD® antibody-phages were divided into three pools corresponding to different VH master genes (pool 1: VH1/5κ, pool 2: VH3 λκ, pool 3: VH2/4/6 λκ). These pools were individually pre-absorbed on mesothelin-negative CHO-K1 cells for depletion of irrelevant antibody phages and subsequently subjected to 3 rounds of alternating whole cell panning on mesothelin-expressing CHO-A9 and NCI-H226 cells followed by pH-elution. Finally, the remaining antibody phages were used to infect E. coli TG1 cells. After centrifugation the bacterial pellet was resuspended in 2×TY medium, plated on agar plates and incubated overnight at 30° C. The selected clones were then scraped from the plates, phages were rescued and amplified. The second and the third round of selections were performed as the initial one.

[0124] The Fab encoding inserts of the selected HuCAL GOLD® phages were subcloned into the expression vector pMORPH® x9_Fab_FS (Rauchenberger, R., et al., J. Biol. Chem. (2003) 278(40): 38194) to facilitate rapid expression of soluble Fab. The DNA of the selected clones was digested with XbaI and EcoRI thereby cutting out the Fab encoding insert (ompA-VLCL and phoA-Fd), and cloned into the XbaI/EcoRI cut vector pMORPH® x9_Fab_FS. Fab expressed in this vector carry two C-terminal tags (FLAG® and Strep-Tag® II) for detection and purification.

C. Affinity Maturation Affinity Maturation of Selected Fab by Stepwise Exchange of CDR Cassettes

[0125] To increase affinity and biological activity of selected antibody fragments (MF-L, MF-A, MF-J, MF-T and MF-226), L-CDR3 and H-CDR2 regions were optimized in parallel by cassette mutagenesis using trinucleotide directed mutagenesis (Virnekas et al, Nucleic Acids Res. 22(25): 5600-7), while the framework regions were kept constant (WO2006122797). Pannings for selection of high affinity phage displayed Fab fragments were performed either on purified biotinylated recombinant mesothelin (human or murine mesothelin) or directly on mesothelin expressing cell lines (NCI-H226 or OVCAR-3). Combinations of these different panning strategies were also applied throughout the three panning rounds which were performed.

Example 2

Epitope Grouping

[0126] Epitope grouping experiments were performed using Biacore by monitering simultaneous binding of pairs of anti-mesothelin antibodies to immobilized mesothelin. Briefly, the first antibody was covalently immobilized to the sensor chip through primary amine coupling using n-hydroxysuccinamide (NHC) and N-ethyl-N'-dimethylaminopropyl carbodiimide (EDC). Unoccupied binding sites on the surface were then blocked with ethanolamide. Soluble mesothelin was captured on the surface via the immobilized antibody, therefore, the epitope of the capture antibody is blocked for all bound mesothelin molecules. A second antibody was immediately passed over the surface to bind to the immobilized mesothelin. Two antibodies recognizing the same or overlapping epitopes cannot bind to the mesothelin, whereas antibodies with distinct epitopes are able to bind. The antibody surface was regenerated with glycine, pH 2.8, to remove bound proteins and then the process was repeated with other antibodies. All combinations of seven antibodies were tested. Representative results using MF-T and several other antibodies are shown in FIG. 1A. Use of MF-T as the second antibody served as a positive control and anti-FLAG served as a negative control. FIG. 1B depicts a summary of the pairwise binding results for seven anti-mesothelin antibodies in a Venn diagram with circles representing individual epitopes. Overlapping circles represent overlapping epitopes. MF428 competed for binding with all other antibodies tested. MF-J and MF-T bind to distinct epitopes compared to each other and to MF-A, MF-226 and MF-L, which seem to compete for the same epitope region. The commercially available mouse antibody K1 binds to an epitope region distinct from the one recognized by MF-J and MF-T, but seems to share a similar epitope region to MF-A, MF-L and MF-226.

Example 3

Cross-Reactivity to Murine Mesothelin

[0127] Shown in Table 5 are results of Biacore and ELISA studies showing cross-reactivity of antibodies of the invention to murine mesothelin. The kinetic constants k_on and k_off were determined with serial dilutions of the respective purified Fab fragment binding to covalently immobilized human or murine mesothelin using the Biacore 3000 instrument (Biacore, Uppsala, Sweden). Covalent antigen immobilization was achieved by a standard EDC-NHS coupling procedure. Kinetic measurements were done in PBS, pH 7.2 at a flow rate of 20 μl/min using Fab concentration ranging from 1.5-500 nM. Injection time for each concentration was 1 min, followed by 3 min dissociation phase. For regeneration 5 μl 10 mM glycine buffer, pH 1.8 was used. All sensograms were fitted using the BIA evaluation software 3.1 (Biacore).

TABLE-US-00005 TABLE 5 Monovalent anti-mesothelin antibody affinities to human and murine mesothelin (Fab formats) Human mesothelin Murine mesothelin Antibody (Fab) K_D [M] kd [1/s] K_D [M] kd [1/s] MF-226 5.8 × 10^-8 3.8 × 10^-2 1.28 × 10^-6 1.4 × 10^-1 MOR 06626 .sup. 6.7 × 10^-10 1.2 × 10^-3 6.7 × 10^-9 9.8 × 10^-3 MOR 06638 1.6 × 10^-8 6.3 × 10^-3 3.2 × 10^-7 4.0 × 10^-2 MF-A 1.9 × 10^-8 7.9 × 10^-2 6.7 × 10^-7 2.7 × 10^-1 MOR 06657 .sup. 9.5 × 10^-10 5.5 × 10^-3 3.6 × 10^-7 1.6 × 10^-1

Example 4

Invariant Binding to Mesothelin on Different Cancer Cell Lines

[0128] FIG. 2 depicts immunoblots of mesothelin-expressing cell lines generated with anti-mesothelin antibody MF-J (A) and MOR 06635 (B). Briefly, cell extracts were generated by a standard lysis protocol by sonicating the cells for 3 min in the presence of DNAse and RNAse. Cell proteins were separated by SDS-PAGE under denaturing and reducing conditions, blotted onto nitrocellulose membranes and incubated with the appropriate primary antibody (MF-J-IgG or MOR 06635-Fab). Anti-human IgG peroxidase-coupled secondary antibody was used for detection, which was performed with ECL substrate. While only one band appeared when extracts of OVCAR-3 cells were blotted with mesothelin antibodies, multiple bands were observed in CHO-A9 and NCI-H226 cells. This indicates the presence of different isoforms of mesothelin in OVCAR-3, CHO-A9 and NCI-H226 cell lines. Since OVCAR-3 and CHO-A9 express the same, fully spliced transcript variant (Muminova, Z. E., et al., BMC Cancer (2004) 4:19), and SEQ ID 371, the multiple bands must be caused by translational or posttranslational modifications, which might consist in, but are not limited to, for example, differences in glycosylation patterns.

[0129] Table 6 shows that EC₅₀ values obtained by FACS titration of representative affinity matured antibodies of the invention on NCI-H226 and OVCAR-3 cells do not vary significantly for a subset of IgGs (i.e. MOR07265, -6631, -6669, -7111, -6640, -6642) while other IgGs show a more than eight fold higher ECso value on OVCAR-3 than NCI-H226 (i.e. MOR06626, -6638, -6657. -6643). Most notably IgGs MOR07265, -6631, -6635, -6669, -7111, -6640, -6642 are affinity matured derivatives of parental IgG MF-J, indicating that these IgGs bind to a related epitope which is invariably present on OVCAR-3 as well as NCI-H226 cells. Thus these data demonstrate the quality of invariant binding provided in the present invention.

[0130] FACS titration was performed in a 96 well microtiter plate, in which serial dilutions of the primary antibody in a volume of 80 μl of FACS buffer (3% FCS, 0.02% NaN3 in PBS) were mixed with 20 μl of a cell suspension consisting of 10⁶ cells/ml which had been detached with accutase or trypsin/EDTA, and resuspended in FACS buffer. Incubation was performed at 4° C. for 1 hour with agitation. Cells were washed twice with FACS buffer and resuspended in 100 μl/well of anti-human PE conjugate solution in FACS buffer. Incubation and washing was performed as before. Analysis of cell-bound antibodies was done using the FACS Array device. EC₅₀ values were determined from fluorescence medians of duplicates using Prism 4.0 software (GraphPad) applying non-linear regression fit.

TABLE-US-00006 TABLE 6 FACS titration of IgG antibodies on NCI-H226 and OVCAR-3 cells EC₅₀ [nM] x-fold different EC₅₀ on Antibody (IgG) NCI-H226 OVCAR-3 OVCAR-3 vs. NCI-H226 MOR06626 0.44 9.68 22.0 MOR06638 0.19 4.19 22.1 MOR07265 1.11 1.06 1.0 MOR06631 2.02 0.96 0.5 MOR 06669 0.41 1.40 3.4 MOR07111 0.80 1.35 1.7 MOR06640 0.63 0.53 0.8 MOR06642 0.58 0.54 0.9 MOR06657 0.14 0.53 14 MOR06643 0.23 1.86 8.1

Example 5

Binding to Mesothelin in the Presence of Cancer Antigen 125 (CA125)

[0131] FIG. 3 shows that cancer antigen 125 (CA125) binds to mesothelin which is in turn bound to a subset of mesothelin antibodies including MOR06640 and MF-T, while other antibodies, such as MF-226, compete with CA125 for mesothelin binding. Data shown are relative light units (RLU) detected by SECTOR Light Imager (Meso Scale Discovery). Plates were coated with the mesothelin antibody depicted at 15 μg/ml, and washed and blocked after each subsequent incubation. Mesothelin was added at the concentrations indicated and titrated down from 10 μg/ml to 0.08 μg/ml. Plates were subsequently incubated with CA125 (Lee Biosolutions, Cat #150-11, 50 000 U/ml diluted 1:300). Detection was performed with a mouse anti-CA125 antibody and an MSD Sulfo tag (Meso Scale Discovery) labelled anti mouse Fab antibody. An unspecific human control antibody was coated as a control. Further controls included the full assay setup with mesothelin at the highest concentrations tested (10 μg/ml) and omission of either CA125 or the mouse anti-CA125 antibody, or full assay setup without mesothelin. This example shows that antibodies, antigen-binding antibody fragments, or variants thereof, which invariantly bind mesothelin can be identified by in vitro testing.

Example 6

Internalization

[0132] Relative internalization of anti-mesothelin antibodies on CHO-A9 cells is shown in FIG. 4. Briefly, CHO-A9 cells expressing mesothelin protein were labeled with ¹²⁵I-anti-mesothelin antibodies for 2 hours at 0° C., to bind the labeled antibody to cell surface mesothelin. The low temperature inhibited internalization. Unbound antibody was washed away using cold buffer and individual aliquots of labeled cells were placed in a 37° C. water bath to initiate internalization. A time course was run in which triplicate samples were collected at: 0, 15, 30, 45, 60, 75 and 90 minutes. At each time point, samples were centrifuged to pellet cells and the supernatant was collected, which contained antibody that had dissociated from the cells. The cell pellet was then briefly washed with acid (PBS+1% glucose pH1.0) in order to remove cell surface-bound labeled antibody, and then pelleted by centrifugation. The supernatant, containing antibody eluted from the cell surface was collected. The pellet fraction, containing internalized antibody, was collected separately. After completion of the time course, the radioactivity in each of the fractions from all time points was determined using a gamma counter. The percentage of total counts present in the fractions represents the percentage of the antibody that was dissociated, bound to the cell surface or internalized at each time point. In experiments in which a second antibody (goat anti-human IgG Fc, or goat anti-mouse IgG Fc, respectively) was added along with the primary labeled antibody to crosslink and thus stabilize the cell surface-bound antibody, much lower antibody dissociation rates were observed compared to cells only treated with the primary antibody. Correspondingly higher internalization levels were also achieved for all antibodies tested with the second antibody. In the absence of a second antibody, the relatively rapid off-rates of the antibodies, as seen in the Biacore studies, reduced the antibodies' residency time on the cell surface such that internalization was significantly reduced. Therefore, four candidate antibodies were chosen for affinity maturation to obtain progenitor antibodies with reduced dissociation rates.

TABLE-US-00007 TABLE 7 Sequences of the antibodies HCDR1 HCDR2 HCDR3 LCDR1 LCDR2 LCDR3 VH VL VH VL SEQ SEQ SEQ SEQ SEQ SEQ Protein Protein Nucleotide Nucleotide Antibody ID ID ID ID ID ID SEQ ID SEQ ID SEQ ID SEQ ID MF-J 1 31 67 99 129 160 198 241 284 327 MOR 1 32 67 99 129 161 199 242 285 328 07265 MOR 1 32 67 99 129 160 200 243 286 329 06631 MOR 1 33 67 99 129 160 201 244 287 330 06669 MOR 1 31 67 99 129 162 202 245 288 331 07111 MOR 1 31 67 99 129 161 203 246 289 332 06640 MOR 1 31 67 99 129 163 204 247 290 333 06642 MOR 2 34 68 100 130 164 205 248 291 334 06643 MF-226 3 35 69 101 131 165 206 249 292 335 MOR 3 36 69 101 131 165 207 250 293 336 06626 MOR 1 37 67 99 129 160 208 251 294 337 06635 MOR 3 35 69 101 131 166 209 252 295 338 06638 MF-A 4 38 70 102 132 167 210 253 296 339 MOR 4 38 70 102 132 168 211 254 297 340 06657 MF-T 5 39 71 103 133 169 212 255 298 341 MF-L 2 34 68 100 130 170 213 256 299 342 MF-1 6 40 72 104 134 171 214 257 300 343 MF-5 7 41 73 105 135 172 215 258 301 344 MF-8 8 42 74 106 136 173 216 259 302 345 MF-24 9 43 75 107 137 174 217 260 303 346 MF-25 10 44 76 108 138 175 218 261 304 347 MF-27 1 45 77 109 139 176 219 262 305 348 MF-73 11 46 78 110 140 177 220 263 306 349 MF-78 12 47 79 111 141 178 221 264 307 350 MF-84 13 48 80 112 142 179 222 265 308 351 MF-101 14 49 81 113 143 180 223 266 309 352 MF-230 15 50 82 114 144 181 224 267 310 353 MF-236 16 51 83 115 145 182 225 268 311 354 MF-252 17 52 84 116 146 183 226 269 312 355 MF-275 17 53 85 117 147 184 227 270 313 356 MF-423 18 54 86 118 148 185 228 271 314 357 MF-427 19 55 87 119 149 186 229 272 315 358 MF-428 20 56 88 120 150 187 230 273 316 359 MF-C 21 57 89 121 151 188 231 274 317 360 MF-I 22 58 90 102 152 189 232 275 318 361 MF-M 23 59 91 122 153 190 233 276 319 362 MF-P 24 60 92 123 154 191 234 277 320 363 MF-Q 25 61 93 124 155 192 235 278 321 364 MF-S 26 62 94 125 156 193 236 279 322 365 MF-U 27 63 95 126 157 194 237 280 323 366 MF-V 28 64 96 127 158 195 238 281 324 367 MF-W 29 65 97 102 159 196 239 282 325 368 MF-Y 30 66 98 128 155 197 240 283 326 369

Sequence CWU 1

1

371110PRTHomo sapiens 1Gly Tyr Ser Phe Thr Asn Tyr Trp Ile Gly 1 5 10 210PRTHomo sapiens 2Gly Phe Thr Phe Ser Asp Tyr Ala Met Ser 1 5 10 310PRTHomo sapiens 3Gly Tyr Thr Phe Thr Gly Asn Tyr Ile Asn 1 5 10 410PRTHomo sapiens 4Gly Gly Thr Phe Ser Ser Tyr Tyr Phe Ser 1 5 10 510PRTHomo sapiens 5Gly Tyr Ser Phe Thr Ser Tyr Trp Ile Gly 1 5 10 610PRTHomo sapiens 6Gly Gly Ser Ile Ser Asn Tyr Tyr Trp Ser 1 5 10 710PRTHomo sapiens 7Gly Phe Thr Phe Ser Ser Tyr Gly Met Ser 1 5 10 810PRTHomo sapiens 8Gly Phe Thr Phe Ser Asn Ser Trp Met Ser 1 5 10 910PRTHomo sapiens 9Gly Gly Thr Phe Ser Asn Tyr Ser Ile Ser 1 5 10 1010PRTHomo sapiens 10Gly Tyr Thr Phe Thr Ser Tyr Ala Leu His 1 5 10 1112PRTHomo sapiens 11Gly Asp Ser Val Ser Ser Arg Ser Ala Ala Trp Gly 1 5 10 1210PRTHomo sapiens 12Gly Gly Thr Phe Ser Asn Tyr Ser Leu His 1 5 10 1310PRTHomo sapiens 13Gly Phe Thr Phe Ser Ser Tyr Ala Met Asn 1 5 10 1410PRTHomo sapiens 14Gly Tyr Thr Phe Thr Ile Tyr Asp Met His 1 5 10 1512PRTHomo sapiens 15Gly Asp Ser Val Ser Ser Asn Ser Ala Ala Trp Gly 1 5 10 169PRTHomo sapiens 16Gly Tyr Ser Phe Asn Thr Ser Trp Ile 1 5 1712PRTHomo sapiens 17Gly Asp Ser Val Ser Ser Asn Ser Ala Ala Trp Ser 1 5 10 1810PRTHomo sapiens 18Gly Tyr Ser Phe Thr Thr Tyr Trp Ile Gly 1 5 10 1912PRTHomo sapiens 19Gly Asp Ser Val Ser Ser Asn Ser Ala Ser Trp Ser 1 5 10 2010PRTHomo sapiens 20Gly Phe Thr Phe Ser Thr Tyr Ala Met Ser 1 5 10 2110PRTHomo sapiens 21Gly Phe Thr Phe Ser Arg Tyr Ala Ile Ser 1 5 10 2210PRTHomo sapiens 22Gly Gly Thr Phe Ser Ser Tyr Tyr Ile Ser 1 5 10 2310PRTHomo sapiens 23Gly Gly Thr Phe Ser Ser His Ala Ile Ser 1 5 10 2412PRTHomo sapiens 24Gly Asp Ser Val Ser Ser Asn Thr Ala Ala Trp Ser 1 5 10 2512PRTHomo sapiens 25Gly Phe Ser Leu Ser Ser Ser Gly Val Gly Val Ser 1 5 10 2612PRTHomo sapiens 26Gly Asp Ser Val Ser Ser Ser Ser Ala Ala Trp Ser 1 5 10 279PRTHomo sapiens 27Gly Phe Thr Phe Ser Ser His Tyr Ser 1 5 2810PRTHomo sapiens 28Gly Gly Thr Phe Ser Asn Tyr Thr Ile Ser 1 5 10 299PRTHomo sapiens 29Gly Phe Thr Phe Ser Ser Tyr Tyr Ser 1 5 3010PRTHomo sapiens 30Gly Phe Thr Phe Ser Asn Tyr Gly Ile His 1 5 10 3120PRTHomo sapiens 31Trp Met Gly Val Ile Met Pro Ser Asp Ser Tyr Thr Arg Tyr Ser Pro 1 5 10 15 Ser Phe Gln Gly 20 3220PRTHomo sapiens 32Trp Met Gly Phe Ile Trp Pro Val Asp Ser Trp Thr Gln Tyr Ser Pro 1 5 10 15 Ser Phe Gln Gly 20 3320PRTHomo sapiens 33Trp Met Gly Ile Ile Trp Pro Ile Asp Ser Phe Thr Gln Tyr Ser Pro 1 5 10 15 Ser Phe Gln Gly 20 3420PRTHomo sapiens 34Trp Val Ser Ala Ile Met Tyr Asp Ser Ser Ser Thr Phe Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 3520PRTHomo sapiens 35Trp Met Gly Ile Ile Asn Pro His Gly Gly Asp Thr Lys Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 3620PRTHomo sapiens 36Trp Met Gly Ile Ile Asn Pro Thr Lys Gly Trp Thr Leu Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 3720PRTHomo sapiens 37Trp Met Gly Ile Ile Asn Pro Thr Lys Gly Trp Thr Leu Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 3820PRTHomo sapiens 38Trp Met Gly Gly Ile Ile Pro Lys Phe Gly Ser Ala Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 3920PRTHomo sapiens 39Trp Met Gly Ile Ile Asp Pro Gly Asp Ser Arg Thr Arg Tyr Ser Pro 1 5 10 15 Ser Phe Gln Gly 20 4019PRTHomo sapiens 40Trp Ile Gly Glu Ile Tyr His Ser Gly Gly Thr Tyr Tyr Asn Pro Ser 1 5 10 15 Leu Lys Gly 4120PRTHomo sapiens 41Trp Val Ser Gly Ile Ser Tyr Ser Ser Ser Ala Thr Tyr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 4220PRTHomo sapiens 42Trp Val Ser Ala Ile Thr Tyr Trp Gly Ser Asn Thr Tyr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 4320PRTHomo sapiens 43Trp Met Gly Arg Ile Ile Pro Asn Phe Gly Thr Ala Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 4420PRTHomo sapiens 44Trp Met Gly Ile Ile Asn Pro Gln Asn Gly Gly Thr Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 4520PRTHomo sapiens 45Trp Met Gly Ile Ile Asp Pro Arg Glu Ser Phe Thr Arg Tyr Ser Pro 1 5 10 15 Ser Phe Gln Gly 20 4621PRTHomo sapiens 46Trp Leu Gly Arg Ile Gly Tyr Arg Ser Lys Trp Met Asn Asp Tyr Ala 1 5 10 15 Val Ser Val Lys Ser 20 4720PRTHomo sapiens 47Trp Met Gly Gly Ile Val Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 4820PRTHomo sapiens 48Trp Val Ser Gly Ile Ser Gly Asn Gly Ser Asn Thr Tyr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 4920PRTHomo sapiens 49Trp Met Gly Tyr Ile Ser Pro Tyr Ser Gly Asp Thr Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 5021PRTHomo sapiens 50Trp Leu Gly Arg Ile Ser Tyr Arg Ser Arg Trp Tyr Asn Asn Tyr Ala 1 5 10 15 Val Ser Val Lys Ser 20 5120PRTHomo sapiens 51Trp Met Gly Ile Ile His Pro Gly His Ser Tyr Thr Arg Tyr Ser Pro 1 5 10 15 Ser Phe Gln Gly 20 5222PRTHomo sapiens 52Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Lys Lys Trp Tyr Asn Asp Tyr 1 5 10 15 Ala Val Ser Val Lys Ser 20 5321PRTHomo sapiens 53Trp Leu Gly Arg Ile Gln Tyr Arg Ser Lys Trp Tyr Asn Ala Tyr Ala 1 5 10 15 Val Ser Val Lys Ser 20 5420PRTHomo sapiens 54Trp Met Gly Phe Ile Tyr Pro Asp Lys Ser Tyr Thr Asn Tyr Ser Pro 1 5 10 15 Ser Phe Gln Gly 20 5521PRTHomo sapiens 55Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Gln Trp Tyr Asn Asp Tyr Ala 1 5 10 15 Val Ser Val Lys Ser 20 5620PRTHomo sapiens 56Trp Val Ser Phe Ile Ser Gly Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 5720PRTHomo sapiens 57Trp Val Ser Ser Ile Ser Gly Gly Gly Ser Lys Thr Phe Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 5820PRTHomo sapiens 58Trp Met Gly Gly Ile Ile Pro Lys Phe Gly Thr Ala Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 5920PRTHomo sapiens 59Trp Met Gly Asn Ile Met Pro Ile Phe Gly Val Ala Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 6021PRTHomo sapiens 60Trp Leu Gly Arg Ile Arg Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 1 5 10 15 Val Ser Val Lys Ser 20 6119PRTHomo sapiens 61Trp Leu Ala Leu Ile Asp Trp Asp Asp Asp Lys Ser Tyr Ser Thr Ser 1 5 10 15 Leu Lys Thr 6221PRTHomo sapiens 62Trp Leu Gly Arg Ile Gly Gln Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 1 5 10 15 Val Ser Val Lys Ser 20 6320PRTHomo sapiens 63Trp Val Ser Thr Ile Ser Ser Asn Gly Ser Tyr Thr Tyr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 6420PRTHomo sapiens 64Trp Met Gly Asn Ile Ile Pro Ala Phe Gly Tyr Ala Asn Tyr Ala Gln 1 5 10 15 Lys Phe Gln Gly 20 6520PRTHomo sapiens 65Trp Val Ser Asn Ile Ser Gly Asn Gly Ser Ser Thr Tyr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 6620PRTHomo sapiens 66Trp Val Ser Tyr Ile Arg Ser Gly Ser Ser Asp Thr Tyr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 6712PRTHomo sapiens 67Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val 1 5 10 6812PRTHomo sapiens 68Ile Asn Tyr Ile Tyr Lys Gly Val His Phe Asp Tyr 1 5 10 6910PRTHomo sapiens 69Trp His His Gly Thr Trp Ile Phe Asp Tyr 1 5 10 706PRTHomo sapiens 70Arg Thr Ser Met Asp Tyr 1 5 7111PRTHomo sapiens 71Gly Gln Leu Tyr Gly Gly Thr Tyr Met Asp Gly 1 5 10 728PRTHomo sapiens 72Pro Met Asp Asn Leu Pro Asp Ile 1 5 737PRTHomo sapiens 73Tyr Leu Tyr Tyr Phe Asp Val 1 5 746PRTHomo sapiens 74Thr Lys Phe Phe Ala Asn 1 5 756PRTHomo sapiens 75Gly Ile Tyr Phe Ala Phe 1 5 7611PRTHomo sapiens 76Lys His Lys Tyr Arg Ile Gly Ser Met Asp Val 1 5 10 778PRTHomo sapiens 77Tyr Met Lys Gly Gly Tyr Asp Tyr 1 5 788PRTHomo sapiens 78Met Gln Gly Phe Gln Leu Asp Tyr 1 5 796PRTHomo sapiens 79Thr Tyr Thr Phe Ala Val 1 5 807PRTHomo sapiens 80Lys Trp Leu Phe Tyr Asp Tyr 1 5 815PRTHomo sapiens 81Gly Trp Gln Asp Phe 1 5 8214PRTHomo sapiens 82Tyr Tyr Ser Asp His Phe Gly Leu Tyr Pro Tyr Phe Asp Tyr 1 5 10 8318PRTHomo sapiens 83Gly Asp Gly Gly Pro Ser Ser Gln Gly Asn Tyr Phe Gly Trp Val Tyr 1 5 10 15 Asp Val 8412PRTHomo sapiens 84Asn Tyr Ser Gly Pro Met Tyr Tyr Tyr Gly Asp Val 1 5 10 8511PRTHomo sapiens 85Gly Phe His Gly Ser Thr Met Tyr Phe Asp Val 1 5 10 868PRTHomo sapiens 86Gly Leu Gly Gly Ser Phe Asp Val 1 5 8711PRTHomo sapiens 87Gly Trp Ile Thr Gly Trp Arg Ile Phe Asp Tyr 1 5 10 8811PRTHomo sapiens 88Lys Met Tyr Trp Trp Ser Asp Gly Phe Asp Tyr 1 5 10 8912PRTHomo sapiens 89Tyr Pro Gly Pro Thr Gly His Val Phe Phe Asp Ile 1 5 10 9012PRTHomo sapiens 90Tyr Pro Gly Pro Thr Gly His Val Phe Phe Asp Ile 1 5 10 916PRTHomo sapiens 91Glu Met Arg Leu Ala Tyr 1 5 9214PRTHomo sapiens 92Gly Tyr His Gln Gly Leu Tyr Gly Asn His Met Phe Asp Val 1 5 10 938PRTHomo sapiens 93Ile Gln Gly Trp Asn Tyr Asp Val 1 5 948PRTHomo sapiens 94Ser Arg Phe Gly Tyr Phe Asp Val 1 5 9510PRTHomo sapiens 95Phe Val Ala Arg Leu Asn Val Phe Asp Tyr 1 5 10 966PRTHomo sapiens 96Lys Phe Thr Phe Asp Val 1 5 9710PRTHomo sapiens 97Leu Ile Ala Thr Leu Gly Thr Phe Asp Tyr 1 5 10 9811PRTHomo sapiens 98Thr Ala Pro Ala Gly His Gly Val Phe Ala Asn 1 5 10 9912PRTHomo sapiens 99Arg Ala Ser Gln Ser Val Arg Ser Ser Arg Leu Ala 1 5 10 10011PRTHomo sapiens 100Arg Ala Ser Gln Ile Val Ser Gly Tyr Leu Ala 1 5 10 10113PRTHomo sapiens 101Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Ser 1 5 10 10214PRTHomo sapiens 102Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser 1 5 10 10314PRTHomo sapiens 103Thr Gly Thr Ser Ser Asp Ile Gly Gly Tyr Asn Ser Val Ser 1 5 10 10411PRTHomo sapiens 104Arg Ala Ser Gln Asn Ile Gly Ser Tyr Leu Asn 1 5 10 10514PRTHomo sapiens 105Thr Gly Thr Ser Ser Asp Val Gly Ala Ser Asp Thr Val Thr 1 5 10 10616PRTHomo sapiens 106Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Thr Tyr Leu Ser 1 5 10 15 10713PRTHomo sapiens 107Ser Gly Ser Ser Ser Asn Ile Gly Pro Asn Tyr Val Ser 1 5 10 10812PRTHomo sapiens 108Arg Ala Ser Gln Ser Leu Thr Ser Asn Gln Leu Ala 1 5 10 10914PRTHomo sapiens 109Thr Gly Thr Ser Ser Asp Val Gly Gly Asn Asn Phe Val Ser 1 5 10 11011PRTHomo sapiens 110Arg Ala Ser Gln Thr Ile Ser Ser Ala Leu Ala 1 5 10 11114PRTHomo sapiens 111Thr Gly Thr Ser Ser Asp Val Gly Asn Phe Asn Tyr Val Asn 1 5 10 11212PRTHomo sapiens 112Arg Ala Ser Gln Ser Val Thr Ser Asn Tyr Leu Ala 1 5 10 11311PRTHomo sapiens 113Arg Ala Ser Gln Ser Ile Asn Arg Ser Leu Thr 1 5 10 11414PRTHomo sapiens 114Thr Gly Thr Ser Ser Asp Ile Gly Gly Phe Asn Tyr Val Ser 1 5 10 11511PRTHomo sapiens 115Ser Gly Asp Asn Ile Pro Asn Phe Tyr Val His 1 5 10 11614PRTHomo sapiens 116Thr Gly Thr Ser Ser Asp Ile Gly Arg Tyr His Tyr Val Ser 1 5 10 11713PRTHomo sapiens 117Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Ser Val Asn 1 5 10 11814PRTHomo sapiens 118Thr Gly Thr Ser Ser Asp Val Gly Asp Tyr Asn Tyr Val Ser 1 5 10 11914PRTHomo sapiens 119Thr Gly Thr Ser Ser Asp Val Gly Asp Tyr Asn Tyr Val Ser 1 5 10 12011PRTHomo sapiens 120Ser Gly Asp Asn Leu Arg Ser Lys Tyr Ala His 1 5 10 12111PRTHomo sapiens 121Ser Gly Asp Asn Ile Gly Ser Lys Val Ala Thr 1 5 10 12214PRTHomo sapiens 122Thr Gly Thr Ser Ser Asp Ile Gly His Phe Asn Tyr Val Ser 1 5 10 12313PRTHomo sapiens 123Ser Gly Ser Ser Ser Asn Ile Gly Ser His Thr Val Asn 1 5 10 12413PRTHomo sapiens 124Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Arg Val Ser 1 5 10 12514PRTHomo sapiens 125Thr Gly Thr Ser Ser Asp Ile Gly Thr Tyr Asn His Val Ser 1 5 10 12614PRTHomo sapiens 126Thr Gly Ser Ser Ser Asn Ile Gly Ile Gly Tyr Asp Val Asn 1 5 10 12714PRTHomo sapiens 127Thr Gly Thr Ser Ser Asp Leu Gly Gly Tyr Ser Tyr Val Ser 1 5 10 12813PRTHomo sapiens 128Ser Gly Ser Ser Ser Asn Ile Gly Asn Tyr Arg Val Ser 1 5 10 12911PRTHomo sapiens 129Leu Leu Ile Tyr Gly Ala Ser Lys Arg Ala Thr 1 5 10 13011PRTHomo sapiens 130Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr 1 5 10 13111PRTHomo sapiens 131Leu Leu Ile Tyr Asn Asp Asn Gln Arg Pro Ser 1 5 10 13211PRTHomo sapiens 132Leu Met Ile Tyr Ser Val Ser Lys Arg Pro Ser 1 5 10 13311PRTHomo sapiens 133Leu Met Ile Tyr Gly Val Asn Asn Arg Pro Ser 1 5 10 13411PRTHomo sapiens 134Leu Leu Ile Tyr Asn Ser Ser Thr Leu Gln Ser 1 5 10 13511PRTHomo sapiens 135Leu Met Ile Tyr Ala Val Asn Lys Arg Pro Ser 1 5 10 13611PRTHomo sapiens 136Leu Leu Ile Tyr Leu Gly Ser Lys Arg Ala Ser 1 5 10 13711PRTHomo sapiens 137Leu Leu Ile His Gly Asn Ala Asn Arg Pro Ser 1 5 10 13811PRTHomo sapiens 138Leu Leu Ile Tyr Asp Ser Ser Asn Arg Ala Thr 1 5 10 13911PRTHomo sapiens 139Leu Met Ile Tyr Tyr Gly Asp Ser Arg Pro Ser 1 5 10 14011PRTHomo sapiens 140Leu Leu Ile Tyr Gly Ala Ser Thr Leu Gln Ser 1 5 10 14111PRTHomo sapiens 141Leu Met Ile Tyr Ser Val Ser Ser Arg Pro Ser 1 5 10 14211PRTHomo sapiens 142Leu Leu Ile Tyr Gly Ser Ser Ser Arg Ala Thr 1

5 10 14311PRTHomo sapiens 143Leu Leu Ile Tyr Ala Ala Ser Asn Leu Gln Ser 1 5 10 14411PRTHomo sapiens 144Leu Met Ile Tyr Asp Val Ser Asn Arg Pro Ser 1 5 10 14511PRTHomo sapiens 145Leu Val Ile Tyr Glu Asp Ser Asp Arg Pro Ser 1 5 10 14611PRTHomo sapiens 146Val Met Ile Tyr Ser Val Ser Lys Arg Pro Ser 1 5 10 14710PRTHomo sapiens 147Leu Leu Ile Tyr Asn Asn Gln Arg Pro Ser 1 5 10 14811PRTHomo sapiens 148Leu Met Ile Tyr Arg Val Asp Asn Arg Pro Ser 1 5 10 14911PRTHomo sapiens 149Leu Leu Ile Tyr Ser Asn Ser Gln Arg Pro Ser 1 5 10 15011PRTHomo sapiens 150Leu Val Ile Tyr Ser Lys Asp Asn Arg Pro Ser 1 5 10 15111PRTHomo sapiens 151Leu Val Ile Tyr Tyr Asp Asn Asp Arg Pro Ser 1 5 10 15211PRTHomo sapiens 152Leu Met Ile Tyr Ser Val Asn Asn Arg Pro Ser 1 5 10 15311PRTHomo sapiens 153Leu Met Ile Tyr Ser Val Ile Ser Arg Pro Ser 1 5 10 15411PRTHomo sapiens 154Leu Leu Ile Tyr Asp Val Asn Lys Arg Pro Ser 1 5 10 15511PRTHomo sapiens 155Leu Leu Ile Tyr Gly Asn Asn Lys Arg Pro Ser 1 5 10 15611PRTHomo sapiens 156Leu Met Ile Tyr Asn Val Asn Lys Arg Pro Ser 1 5 10 15711PRTHomo sapiens 157Leu Leu Ile Tyr Lys Asn Thr Asn Arg Pro Ser 1 5 10 15811PRTHomo sapiens 158Leu Met Ile Tyr Asn Val Asn Asn Arg Pro Ser 1 5 10 15911PRTHomo sapiens 159Val Met Ile Tyr Gly Val Thr Lys Arg Pro Ser 1 5 10 1608PRTHomo sapiens 160Gln Gln Tyr Tyr Asp Phe Pro Pro 1 5 1619PRTHomo sapiens 161Gln Gln Tyr Ser His Asp Pro Ser Gly 1 5 1628PRTHomo sapiens 162Gln Gln Phe Tyr Ser Lys Pro Ile 1 5 1639PRTHomo sapiens 163Gln Gln Tyr Ser Gln Asp Pro Ser Ser 1 5 1649PRTHomo sapiens 164Gln Gln Trp Ser Leu Arg Ser Pro Phe 1 5 1659PRTHomo sapiens 165Ser Thr Tyr Asp Arg Arg Thr Phe Ser 1 5 16610PRTHomo sapiens 166Ser Ser Trp Asp Arg Ala Asp Gly Ser Tyr 1 5 10 16710PRTHomo sapiens 167Gly Ala Trp Ala His Met Ser Leu Gly Lys 1 5 10 16810PRTHomo sapiens 168Ala Thr Trp Asp His Ser Gln Met Gly Lys 1 5 10 16910PRTHomo sapiens 169Ser Ser Tyr Asp Ile Glu Ser Ala Thr Pro 1 5 10 1708PRTHomo sapiens 170Gln Gln Tyr Tyr Asn Phe Ser Phe 1 5 1718PRTHomo sapiens 171Gln Gln Arg Ser Asn Met Pro Ile 1 5 1728PRTHomo sapiens 172Ala Ser Arg Asp Ser Ser Ser Met 1 5 1738PRTHomo sapiens 173Gln Gln Tyr Tyr Asp Ser Ser Ser 1 5 17410PRTHomo sapiens 174Gln Ser Tyr Asp Phe Phe Thr Asn Ser Ser 1 5 10 1758PRTHomo sapiens 175Gln Gln Tyr Gly Ser Phe Pro Ala 1 5 17610PRTHomo sapiens 176Gln Ser Trp Asp Ala Pro Met Gly Met Trp 1 5 10 1778PRTHomo sapiens 177Gln Gln Tyr Tyr Ser Tyr Ser Val 1 5 1789PRTHomo sapiens 178Gly Ala Tyr Thr Thr Asp Thr Leu Ser 1 5 1798PRTHomo sapiens 179Gln Gln Tyr Ser Asp Ile Pro Ala 1 5 1808PRTHomo sapiens 180Leu Gln Val Tyr Asn Leu Pro Leu 1 5 18110PRTHomo sapiens 181Ser Thr Tyr Asp Asp Asp Gln Gln Asp Ala 1 5 10 18210PRTHomo sapiens 182Gln Ser Tyr Asp Lys Pro Thr Phe Ser Gly 1 5 10 18310PRTHomo sapiens 183Ser Ala Tyr Asp Thr Asn Asn Tyr Leu Ser 1 5 10 1847PRTHomo sapiens 184Gln Ala Tyr Ala Ser Asn Ile 1 5 1858PRTHomo sapiens 185Gln Ser Trp Val Gly Pro Ser Thr 1 5 1869PRTHomo sapiens 186Gln Ser Tyr Asp His Asn Ser Tyr Thr 1 5 1878PRTHomo sapiens 187Ser Ser Trp Ala His Asp His Lys 1 5 18810PRTHomo sapiens 188Gln Ser Tyr Asp Gly Gln Met Ser Thr Ser 1 5 10 18910PRTHomo sapiens 189Ser Thr Tyr Asp His Thr Ser Ser Gly Phe 1 5 10 1908PRTHomo sapiens 190Ala Ser Phe Thr Phe Pro Ser Leu 1 5 19110PRTHomo sapiens 191Ala Ser Trp Asp Ser Val Gln Val Ser Pro 1 5 10 1929PRTHomo sapiens 192Ser Ala Trp Asp Leu Leu Glu Val Tyr 1 5 1938PRTHomo sapiens 193Ser Ala Tyr Ala Pro Ser Ala Val 1 5 19410PRTHomo sapiens 194Ser Ser Tyr Asp Ser Phe His His Gly Ile 1 5 10 19510PRTHomo sapiens 195Ser Ser Tyr Asp Tyr Val Ser Ser Asp Thr 1 5 10 19610PRTHomo sapiens 196Gln Thr Tyr Asp Glu His Gly Phe His Ile 1 5 10 19710PRTHomo sapiens 197Ser Thr Tyr Thr Gly Leu Pro Phe Thr Thr 1 5 10 198121PRTHomo sapiens 198Gln Val Glu Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Met Pro Ser Asp Ser Tyr Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 199121PRTHomo sapiens 199Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Phe Ile Trp Pro Val Asp Ser Trp Thr Gln Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 200121PRTHomo sapiens 200Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Phe Ile Trp Pro Val Asp Ser Trp Thr Gln Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 201121PRTHomo sapiens 201Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Trp Pro Ile Asp Ser Phe Thr Gln Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 202121PRTHomo sapiens 202Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Met Pro Ser Asp Ser Tyr Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 203121PRTHomo sapiens 203Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Met Pro Ser Asp Ser Tyr Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 204121PRTHomo sapiens 204Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Met Pro Ser Asp Ser Tyr Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 205121PRTHomo sapiens 205Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Met Tyr Asp Ser Ser Ser Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ile Asn Tyr Ile Tyr Lys Gly Val His Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 206119PRTHomo sapiens 206Gln Val Glu Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Asn 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Asn Pro His Gly Gly Asp Thr Lys Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp His His Gly Thr Trp Ile Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 207119PRTHomo sapiens 207Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Asn 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Asn Pro Thr Lys Gly Trp Thr Leu Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp His His Gly Thr Trp Ile Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 208121PRTHomo sapiens 208Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Phe Ile Trp Pro Ser Asp Ser Trp Thr Ser Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly His Gly Met Tyr Gly Gly Ala Leu Asp Val Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 209119PRTHomo sapiens 209Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Asn 20 25 30 Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Asn Pro His Gly Gly Asp Thr Lys Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp His His Gly Thr Trp Ile Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 210115PRTHomo sapiens 210Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Tyr Phe Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Lys Phe Gly Ser Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Arg Thr Ser Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105

110 Val Ser Ser 115 211115PRTHomo sapiens 211Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Tyr Phe Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Lys Phe Gly Ser Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Arg Thr Ser Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 212120PRTHomo sapiens 212Gln Val Glu Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Asp Pro Gly Asp Ser Arg Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Gly Gln Leu Tyr Gly Gly Thr Tyr Met Asp Gly Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 213121PRTHomo sapiens 213Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Met Tyr Asp Ser Ser Ser Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ile Asn Tyr Ile Tyr Lys Gly Val His Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 214116PRTHomo sapiens 214Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Gly Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asn Tyr 20 25 30 Tyr Trp Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr His Ser Gly Gly Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60 Gly Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 65 70 75 80 Lys Leu Ser Ser Val Thr Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Pro Met Asp Asn Leu Pro Asp Ile Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 215116PRTHomo sapiens 215Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Tyr Ser Ser Ser Ala Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Leu Tyr Tyr Phe Asp Val Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 216115PRTHomo sapiens 216Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ser 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Thr Tyr Trp Gly Ser Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Lys Phe Phe Ala Asn Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 217115PRTHomo sapiens 217Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Ser Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Ile Pro Asn Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Ile Tyr Phe Ala Phe Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 218120PRTHomo sapiens 218Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Leu His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Asn Pro Gln Asn Gly Gly Thr Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Lys His Lys Tyr Arg Ile Gly Ser Met Asp Val Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 219117PRTHomo sapiens 219Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Asp Pro Arg Glu Ser Phe Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Tyr Met Lys Gly Gly Tyr Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 220120PRTHomo sapiens 220Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Arg 20 25 30 Ser Ala Ala Trp Gly Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Ile Gly Tyr Arg Ser Lys Trp Met Asn Asp Tyr Ala 50 55 60 Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Ala Arg Met Gln Gly Phe Gln Leu Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 221115PRTHomo sapiens 221Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Ser Leu His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Val Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Tyr Thr Phe Ala Val Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 222116PRTHomo sapiens 222Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Gly Asn Gly Ser Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Lys Trp Leu Phe Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 223114PRTHomo sapiens 223Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ile Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Ser Pro Tyr Ser Gly Asp Thr Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Trp Gln Asp Phe Trp Gly Gln Gly Thr Leu Val Thr Val 100 105 110 Ser Ser 224126PRTHomo sapiens 224Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30 Ser Ala Ala Trp Gly Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Ile Ser Tyr Arg Ser Arg Trp Tyr Asn Asn Tyr Ala 50 55 60 Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Ala Arg Tyr Tyr Ser Asp His Phe Gly Leu Tyr Pro Tyr 100 105 110 Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 225126PRTHomo sapiens 225Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Asn Thr Ser 20 25 30 Trp Ile Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met Gly 35 40 45 Ile Ile His Pro Gly His Ser Tyr Thr Arg Tyr Ser Pro Ser Phe Gln 50 55 60 Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr Leu 65 70 75 80 Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Arg Gly Asp Gly Gly Pro Ser Ser Gln Gly Asn Tyr Phe Gly Trp Val 100 105 110 Tyr Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 226125PRTHomo sapiens 226Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30 Ser Ala Ala Trp Ser Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Lys Lys Trp Tyr Asn Asp Tyr 50 55 60 Ala Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys 65 70 75 80 Asn Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala 85 90 95 Val Tyr Tyr Cys Ala Arg Asn Tyr Ser Gly Pro Met Tyr Tyr Tyr Gly 100 105 110 Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 227123PRTHomo sapiens 227Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30 Ser Ala Ala Trp Ser Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Ile Gln Tyr Arg Ser Lys Trp Tyr Asn Ala Tyr Ala 50 55 60 Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Ala Arg Gly Phe His Gly Ser Thr Met Tyr Phe Asp Val 100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 228117PRTHomo sapiens 228Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Thr Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Phe Ile Tyr Pro Asp Lys Ser Tyr Thr Asn Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Gly Leu Gly Gly Ser Phe Asp Val Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 229123PRTHomo sapiens 229Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30 Ser Ala Ser Trp Ser Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Gln Trp Tyr Asn Asp Tyr Ala 50 55 60 Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys

Ala Arg Gly Trp Ile Thr Gly Trp Arg Ile Phe Asp Tyr 100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 230120PRTHomo sapiens 230Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Phe Ile Ser Gly Tyr Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Lys Met Tyr Trp Trp Ser Asp Gly Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 231121PRTHomo sapiens 231Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Gly Gly Gly Ser Lys Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Pro Gly Pro Thr Gly His Val Phe Phe Asp Ile Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 232116PRTHomo sapiens 232Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Tyr Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Lys Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Ile Gly Ile Tyr Asp Ser Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 233115PRTHomo sapiens 233Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser His 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Met Pro Ile Phe Gly Val Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Met Arg Leu Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 234126PRTHomo sapiens 234Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30 Thr Ala Ala Trp Ser Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Ile Arg Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60 Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Ala Arg Gly Tyr His Gln Gly Leu Tyr Gly Asn His Met 100 105 110 Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 235118PRTHomo sapiens 235Gln Val Gln Leu Lys Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Ser Ser 20 25 30 Gly Val Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Leu Ile Asp Trp Asp Asp Asp Lys Ser Tyr Ser Thr Ser 50 55 60 Leu Lys Thr Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ile Gln Gly Trp Asn Tyr Asp Val Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 236120PRTHomo sapiens 236Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Ser 20 25 30 Ser Ala Ala Trp Ser Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu 35 40 45 Trp Leu Gly Arg Ile Gly Gln Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60 Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn 65 70 75 80 Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Ala Arg Ser Arg Phe Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 237118PRTHomo sapiens 237Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30 Tyr Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 35 40 45 Thr Ile Ser Ser Asn Gly Ser Tyr Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Phe Val Ala Arg Leu Asn Val Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 238115PRTHomo sapiens 238Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Thr Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Asn Ile Ile Pro Ala Phe Gly Tyr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Lys Phe Thr Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115 239118PRTHomo sapiens 239Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Tyr Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 35 40 45 Asn Ile Ser Gly Asn Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Leu Ile Ala Thr Leu Gly Thr Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 240120PRTHomo sapiens 240Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Arg Ser Gly Ser Ser Asp Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Ala Pro Ala Gly His Gly Val Phe Ala Asn Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 241110PRTHomo sapiens 241Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Asp Phe Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 242111PRTHomo sapiens 242Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ser His Asp Pro 85 90 95 Ser Gly Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 243110PRTHomo sapiens 243Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Asp Phe Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 244110PRTHomo sapiens 244Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Asp Phe Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 245110PRTHomo sapiens 245Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Phe Tyr Ser Lys Pro 85 90 95 Ile Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 246111PRTHomo sapiens 246Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ser His Asp Pro 85 90 95 Ser Gly Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 247111PRTHomo sapiens 247Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ser Gln Asp Pro 85 90 95 Ser Ser Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 248110PRTHomo sapiens 248Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ile Val Ser Gly Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Leu Arg Ser Pro 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 249111PRTHomo sapiens 249Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile

Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Asn Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Thr Tyr Asp Arg Arg Thr 85 90 95 Phe Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 250111PRTHomo sapiens 250Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Asn Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Thr Tyr Asp Arg Arg Thr 85 90 95 Phe Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 251110PRTHomo sapiens 251Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Ser 20 25 30 Arg Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Lys Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Asp Phe Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 252112PRTHomo sapiens 252Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Asn Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Trp Asp Arg Ala Asp 85 90 95 Gly Ser Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 253113PRTHomo sapiens 253Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Ser Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Ala Trp Ala His Met 85 90 95 Ser Leu Gly Lys Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 254113PRTHomo sapiens 254Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Ser Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Trp Asp His Ser 85 90 95 Gln Met Gly Lys Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 255224PRTHomo sapiens 255Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Gly Tyr 20 25 30 Asn Ser Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Gly Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Asp Ile Glu 85 90 95 Ser Ala Thr Pro Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly 115 120 125 Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser 130 135 140 Asn Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 145 150 155 160 Leu Ile Tyr Asn Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe 165 170 175 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 180 185 190 Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Thr Tyr Asp Arg Arg 195 200 205 Thr Phe Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 210 215 220 256109PRTHomo sapiens 256Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ile Val Ser Gly Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Asn Phe Ser Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 257109PRTHomo sapiens 257Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Gly Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asn Ser Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Ser Asn Met Pro Ile 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 258111PRTHomo sapiens 258Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ala Ser 20 25 30 Asp Thr Val Thr Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Ala Val Asn Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Arg Asp Ser Ser 85 90 95 Ser Met Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 259114PRTHomo sapiens 259Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 20 25 30 Asn Gly Tyr Thr Tyr Leu Ser Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Lys Arg Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Tyr 85 90 95 Tyr Asp Ser Ser Ser Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 Arg Thr 260112PRTHomo sapiens 260Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Pro Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile His Gly Asn Ala Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Phe Phe Thr 85 90 95 Asn Ser Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 261110PRTHomo sapiens 261Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Leu Thr Ser Asn 20 25 30 Gln Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Asp Ser Ser Asn Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Phe Pro 85 90 95 Ala Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 262113PRTHomo sapiens 262Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Asn 20 25 30 Asn Phe Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Tyr Gly Asp Ser Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Trp Asp Ala Pro 85 90 95 Met Gly Met Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 263109PRTHomo sapiens 263Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Ile Ser Ser Ala 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Ser Tyr Ser Val 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 264112PRTHomo sapiens 264Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Asn Phe 20 25 30 Asn Tyr Val Asn Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Ser Val Ser Ser Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Ala Tyr Thr Thr Asp 85 90 95 Thr Leu Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 265110PRTHomo sapiens 265Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Thr Ser Asn 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ser Ser Ser Arg Ala Thr Gly Val Pro Ala Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ser Asp Ile Pro 85 90 95 Ala Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 110 266109PRTHomo sapiens 266Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Arg Ser 20 25 30 Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Asn Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Leu Gln Val Tyr Asn Leu Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 267109PRTHomo sapiens 267Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Arg Ser 20 25 30 Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Asn Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Leu Gln Val Tyr Asn Leu Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 100 105 268110PRTHomo sapiens 268Asp Ile Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln 1 5 10 15 Thr Ala Arg Ile Ser Cys Ser Gly Asp Asn Ile Pro Asn Phe Tyr Val 20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Glu Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Lys Pro Thr Phe Ser 85 90 95 Gly Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 269113PRTHomo sapiens 269Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Arg Tyr 20 25 30 His Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Val 35 40 45 Met Ile Tyr Ser Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ala Tyr Asp Thr Asn 85 90 95 Asn Tyr Leu Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 270108PRTHomo sapiens 270Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn 20 25 30 Ser Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln Ser 65 70 75 80 Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Tyr Ala Ser Asn Ile Val 85 90 95 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 271111PRTHomo sapiens 271Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Asp Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Arg Val Asp Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Trp Val Gly Pro 85 90 95 Ser Thr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 272111PRTHomo sapiens 272Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ser Asn Ser Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp His Asn Ser 85 90 95 Tyr Thr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 273108PRTHomo sapiens 273Asp Ile Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln 1 5 10 15 Thr Ala Arg Ile Ser Cys Ser Gly Asp Asn Leu Arg Ser Lys Tyr Ala 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Ser Lys Asp Asn Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Trp Ala His Asp His Lys Val 85 90 95 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 274110PRTHomo sapiens 274Asp Ile Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln 1 5 10 15 Thr Ala Arg Ile Ser Cys Ser Gly Asp Asn Ile Gly Ser Lys Val Ala 20 25 30 Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Tyr Asp Asn Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Gly Gln Met Ser Thr 85 90 95 Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 275113PRTHomo sapiens 275Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Ser Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Thr Tyr Asp His Thr 85 90 95 Ser Ser Gly Phe Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 276111PRTHomo sapiens 276Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly His Phe 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Ser Val Ile Ser Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Phe Thr Phe Pro 85 90 95 Ser Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 277112PRTHomo sapiens 277Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser His 20 25 30 Thr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Asp Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Trp Asp Ser Val Gln 85 90 95 Val Ser Pro Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 278111PRTHomo sapiens 278Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn 20 25 30 Arg Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Gly Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ala Trp Asp Leu Leu Glu 85 90 95 Val Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 279111PRTHomo sapiens 279Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Thr Tyr 20 25 30 Asn His Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asn Val Asn Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ala Tyr Ala Pro Ser 85 90 95 Ala Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 280113PRTHomo sapiens 280Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ile Gly 20 25 30 Tyr Asp Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Lys Asn Thr Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 65 70 75 80 Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Asp Ser Phe 85 90 95 His His Gly Ile Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 281113PRTHomo sapiens 281Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Leu Gly Gly Tyr 20 25 30 Ser Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asn Val Asn Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Asp Tyr Val 85 90 95 Ser Ser Asp Thr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 282113PRTHomo sapiens 282Asp Ile Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Val 35 40 45 Met Ile Tyr Gly Val Thr Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Thr Tyr Asp Glu His 85 90 95 Gly Phe His Ile Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln 283112PRTHomo sapiens 283Asp Ile Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Tyr 20 25 30 Arg Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Gly Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Thr Tyr Thr Gly Leu Pro 85 90 95 Phe Thr Thr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 284363DNAHomo sapiens 284caggtggaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcgtt atcatgccgt ctgatagcta tacccgttat 180tctccgagct ttcagggcca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363285363DNAHomo sapiens 285caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcttt atttggcctg ttgattcttg gactcagtat 180tctccttctt ttcagggtca ggtcaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363286363DNAHomo sapiens 286caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcttt atttggcctg ttgattcttg gactcagtat 180tctccttctt ttcagggtca ggtcaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363287363DNAHomo sapiens 287caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcatt atttggccta ttgattcttt tactcagtat 180tctccttctt ttcagggtca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363288363DNAHomo sapiens 288caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcgtt atcatgccgt ctgatagcta tacccgttat 180tctccgagct ttcagggcca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363289363DNAHomo sapiens 289caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcgtt atcatgccgt ctgatagcta tacccgttat 180tctccgagct ttcagggcca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363290363DNAHomo sapiens 290caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcgtt atcatgccgt ctgatagcta tacccgttat 180tctccgagct ttcagggcca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363291363DNAHomo sapiens 291caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct gattatgcta tgtcttgggt gcgccaagcc 120cctgggaagg gtctcgagtg ggtgagcgct atcatgtatg attctagctc taccttttat 180gcggatagcg

tgaaaggccg ttttaccatt tcacgtgata attcgaaaaa caccctgtat 240ctgcaaatga acagcctgcg tgcggaagat acggccgtgt attattgcgc gcgtattaat 300tatatttata agggtgttca ttttgattat tggggccaag gcaccctggt gacggttagc 360tca 363292357DNAHomo sapiens 292caggtggaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60agctgcaaag cctccggata tacctttact ggtaattata ttaattgggt ccgccaagcc 120cctgggcagg gtctcgagtg gatgggcatt atcaatccgc atggtggcga tacgaagtac 180gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgttggcat 300catggtactt ggatttttga ttattggggc caaggcaccc tggtgacggt tagctca 357293357DNAHomo sapiens 293caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60agctgcaaag cctccggata tacctttact ggtaattata ttaattgggt ccgccaagcc 120cctgggcagg gtctcgagtg gatgggcatt attaatccta ctaagggttg gactctttat 180gctcagaagt ttcagggtcg ggtcaccatg acccgtgata ccagcattag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgttggcat 300catggtactt ggatttttga ttattggggc caaggcaccc tggtgacggt tagctca 357294363DNAHomo sapiens 294caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcttt atttggcctt ctgattcttg gacttcttat 180tctccttctt ttcagggtca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatggt 300catggtatgt atggtggtgc tcttgatgtt tggggccaag gcaccctggt gacggttagc 360tca 363295357DNAHomo sapiens 295caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60agctgcaaag cctccggata tacctttact ggtaattata ttaattgggt ccgccaagcc 120cctgggcagg gtctcgagtg gatgggcatt atcaatccgc atggtggcga tacgaagtac 180gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgttggcat 300catggtactt ggatttttga ttattggggc caaggcaccc tggtgacggt tagctca 357296345DNAHomo sapiens 296caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct tcttattatt tttcttgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggcggt atcattccga agtttggctc tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtcgtact 300tctatggatt attggggcca aggcaccctg gtgacggtta gctca 345297345DNAHomo sapiens 297caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct tcttattatt tttcttgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggcggt atcattccga agtttggctc tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtcgtact 300tctatggatt attggggcca aggcaccctg gtgacggtta gctca 345298360DNAHomo sapiens 298caggtggaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact tcttattgga ttggttgggt gcgccaggcc 120cctgggaagg gtctcgagtg gatgggcatt atcgatccgg gtgatagccg tacccgttat 180tctccgagct ttcagggcca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgtggtcag 300ctttatggtg gtacttatat ggatggttgg ggccaaggca ccctggtgac ggttagctca 360299363DNAHomo sapiens 299caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct gattatgcta tgtcttgggt gcgccaagcc 120cctgggaagg gtctcgagtg ggtgagcgct atcatgtatg attctagctc taccttttat 180gcggatagcg tgaaaggccg ttttaccatt tcacgtgata attcgaaaaa caccctgtat 240ctgcaaatga acagcctgcg tgcggaagat acggccgtgt attattgcgc gcgtattaat 300tatatttata agggtgttca ttttgattat tggggccaag gcaccctggt gacggttagc 360tca 363300348DNAHomo sapiens 300caggtgcaat tgcaagaaag tggtccgggc ctggtgaaac cgggcgaaac cctgagcctg 60acctgcaccg tttccggagg cagcatttct aattattatt ggtcttggat tcgccaggcc 120cctgggaagg gtctcgagtg gattggcgag atctatcatt ctggcggtac ctattataat 180ccgagcctga aaggccgggt gaccattagc gttgatactt cgaaaaacca gtttagcctg 240aaactgagca gcgtgacggc ggaagatacg gccgtgtatt attgcgcgcg tcctatggat 300aatcttcctg atatttgggg ccaaggcacc ctggtgacgg ttagctca 348301348DNAHomo sapiens 301caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct tcttatggta tgtcttgggt gcgccaagcc 120cctgggaagg gtctcgagtg ggtgagcggt atctcttatt cttctagcgc tacctattat 180gcggatagcg tgaaaggccg ttttaccatt tcacgtgata attcgaaaaa caccctgtat 240ctgcaaatga acagcctgcg tgcggaagat acggccgtgt attattgcgc gcgttatctt 300tattattttg atgtttgggg ccaaggcacc ctggtgacgg ttagctca 348302345DNAHomo sapiens 302caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct aattcttgga tgtcttgggt gcgccaagcc 120cctgggaagg gtctcgagtg ggtgagcgct atcacttatt ggggtagcaa tacctattat 180gcggatagcg tgaaaggccg ttttaccatt tcacgtgata attcgaaaaa caccctgtat 240ctgcaaatga acagcctgcg tgcggaagat acggccgtgt attattgcgc gcgtactaag 300ttttttgcta attggggcca aggcaccctg gtgacggtta gctca 345303345DNAHomo sapiens 303caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct aattattcta tttcttgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggccgt atcattccga attttggcac tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggtatt 300tattttgctt tttggggcca aggcaccctg gtgacggtta gctca 345304360DNAHomo sapiens 304caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60agctgcaaag cctccggata tacctttact tcttatgctc ttcattgggt ccgccaagcc 120cctgggcagg gtctcgagtg gatgggcatt atcaatccgc agaatggcgg tacgaattac 180gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtaagcat 300aagtatcgta ttggttctat ggatgtttgg ggccaaggca ccctggtgac ggttagctca 360305351DNAHomo sapiens 305caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact aattattgga ttggttgggt gcgccagatg 120cctgggaagg gtctcgagtg gatgggcatt atcgatccgc gtgagagctt tacccgttat 180tctccgagct ttcagggcca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgttatatg 300aagggtggtt atgattattg gggccaaggc accctggtga cggttagctc a 351306360DNAHomo sapiens 306caggtgcaat tgcaacagtc tggtccgggc ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc tctcgttctg ctgcttgggg ttggattcgc 120cagtctcctg ggcgtggcct cgagtggctg ggccgtatcg gttatcgtag caagtggatg 180aacgattatg cggtgagcgt gaaaagccgg attaccatca acccggatac ttcgaaaaac 240cagtttagcc tgcaactgaa cagcgtgacc ccggaagata cggccgtgta ttattgcgcg 300cgtatgcagg gttttcagct tgattattgg ggccaaggca ccctggtgac ggttagctca 360307345DNAHomo sapiens 307caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct aattattctc ttcattgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggcggt atcgttccga tttttggcac tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtacttat 300acttttgctg tttggggcca aggcaccctg gtgacggtta gctca 345308345DNAHomo sapiens 308caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct aattattctc ttcattgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggcggt atcgttccga tttttggcac tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtacttat 300acttttgctg tttggggcca aggcaccctg gtgacggtta gctca 345309342DNAHomo sapiens 309caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgcgag cgtgaaagtg 60agctgcaaag cctccggata tacctttact atttatgata tgcattgggt ccgccaagcc 120cctgggcagg gtctcgagtg gatgggctat atctctccgt attctggcga tacgaattac 180gcgcagaagt ttcagggccg ggtgaccatg acccgtgata ccagcattag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtggttgg 300caggattttt ggggccaagg caccctggtg acggttagct ca 342310378DNAHomo sapiens 310caggtgcaat tgcaacagtc tggtccgggc ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc tctaattctg ctgcttgggg ttggattcgc 120cagtctcctg ggcgtggcct cgagtggctg ggccgtatct cttatcgtag ccgttggtat 180aacaattatg cggtgagcgt gaaaagccgg attaccatca acccggatac ttcgaaaaac 240cagtttagcc tgcaactgaa cagcgtgacc ccggaagata cggccgtgta ttattgcgcg 300cgttattatt ctgatcattt tggtctttat ccttattttg attattgggg ccaaggcacc 360ctggtgacgg ttagctca 378311378DNAHomo sapiens 311caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttaat acttcttgga tttgggtgcg ccagatgcct 120gggaagggtc tcgagtggat gggcattatc catccgggtc atagctatac ccgttattct 180ccgagctttc agggccaggt gaccattagc gcggataaaa gcattagcac cgcgtatctt 240caatggagca gcctgaaagc gagcgatacg gccatgtatt attgcgcgcg tggtgatggt 300ggtccttctt ctcagggtaa ttattttggt tgggtttatg atgtttgggg ccaaggcacc 360ctggtgacgg ttagctca 378312375DNAHomo sapiens 312caggtgcaat tgcaacagtc tggtccgggc ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc tctaattctg ctgcttggtc ttggattcgc 120cagtctcctg ggcgtggcct cgagtggctg ggccgtatct attatcgtag caagaagtgg 180tataacgatt atgcggtgag cgtgaaaagc cggattacca tcaacccgga tacttcgaaa 240aaccagttta gcctgcaact gaacagcgtg accccggaag atacggccgt gtattattgc 300gcgcgtaatt attctggtcc tatgtattat tatggtgatg tttggggcca aggcaccctg 360gtgacggtta gctca 375313369DNAHomo sapiens 313caggtgcaat tgcaacagtc tggtccgggc ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc tctaattctg ctgcttggtc ttggattcgc 120cagtctcctg ggcgtggcct cgagtggctg ggccgtatcc agtatcgtag caagtggtat 180aacgcttatg cggtgagcgt gaaaagccgg attaccatca acccggatac ttcgaaaaac 240cagtttagcc tgcaactgaa cagcgtgacc ccggaagata cggccgtgta ttattgcgcg 300cgtggttttc atggttctac tatgtatttt gatgtttggg gccaaggcac cctggtgacg 360gttagctca 369314351DNAHomo sapiens 314caggtgcaat tggttcagag cggcgcggaa gtgaaaaaac cgggcgaaag cctgaaaatt 60agctgcaaag gttccggata ttcctttact acttattgga ttggttgggt gcgccaagcc 120cctgggaagg gtctcgagtg gatgggcttt atctatccgg ataagagcta taccaattat 180tctccgagct ttcagggcca ggtgaccatt agcgcggata aaagcattag caccgcgtat 240cttcaatgga gcagcctgaa agcgagcgat acggccatgt attattgcgc gcgtggtctt 300ggtggttctt ttgatgtttg gggccaaggc accctggtga cggttagctc a 351315369DNAHomo sapiens 315caggtgcaat tgcaacagtc tggtccgggc ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc tctaattctg cttcttggtc ttggattcgc 120cagtctcctg ggcgtggcct cgagtggctg ggccgtatct attatcgtag ccagtggtat 180aacgattatg cggtgagcgt gaaaagccgg attaccatca acccggatac ttcgaaaaac 240cagtttagcc tgcaactgaa cagcgtgacc ccggaagata cggccgtgta ttattgcgcg 300cgtggttgga ttactggttg gcgtattttt gattattggg gccaaggcac cctggtgacg 360gttagctca 369316360DNAHomo sapiens 316caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct acttatgcta tgtcttgggt gcgccaagcc 120cctgggaagg gtctcgagtg ggtgagcttt atctctggtt atggtagctc tacctattat 180gcggatagcg tgaaaggccg ttttaccatt tcacgtgata attcgaaaaa caccctgtat 240ctgcaaatga acagcctgcg tgcggaagat acggccgtgt attattgcgc gcgtaagatg 300tattggtggt ctgatggttt tgattattgg ggccaaggca ccctggtgac ggttagctca 360317363DNAHomo sapiens 317caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct cgttatgcta tttcttgggt gcgccaagcc 120cctgggaagg gtctcgagtg ggtgagctct atctctggtg gtggtagcaa gaccttttat 180gcggatagcg tgaaaggccg ttttaccatt tcacgtgata attcgaaaaa caccctgtat 240ctgcaaatga acagcctgcg tgcggaagat acggccgtgt attattgcgc gcgttatcct 300ggtcctactg gtcatgtttt ttttgatatt tggggccaag gcaccctggt gacggttagc 360tca 363318348DNAHomo sapiens 318caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct tcttattata tttcttgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggcggt atcattccga agtttggcac tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtactatt 300ggtatttatg attcttgggg ccaaggcacc ctggtgacgg ttagctca 348319345DNAHomo sapiens 319caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct tctcatgcta tttcttgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggcaat atcatgccga tttttggcgt tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtgagatg 300cgtcttgctt attggggcca aggcaccctg gtgacggtta gctca 345320378DNAHomo sapiens 320caggtgcaat tgcaacagtc tggtccgggc ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc tctaatactg ctgcttggtc ttggattcgc 120cagtctcctg ggcgtggcct cgagtggctg ggccgtatcc gttatcgtag caagtggtat 180aacgattatg cggtgagcgt gaaaagccgg attaccatca acccggatac ttcgaaaaac 240cagtttagcc tgcaactgaa cagcgtgacc ccggaagata cggccgtgta ttattgcgcg 300cgtggttatc atcagggtct ttatggtaat catatgtttg atgtttgggg ccaaggcacc 360ctggtgacgg ttagctca 378321354DNAHomo sapiens 321caggtgcaat tgaaagaaag cggcccggcc ctggtgaaac cgacccaaac cctgaccctg 60acctgtacct tttccggatt tagcctgtct tcttctggtg ttggtgtgtc ttggattcgc 120cagccgcctg ggaaagccct cgagtggctg gctcttatcg attgggatga tgataagtct 180tatagcacca gcctgaaaac gcgtctgacc attagcaaag atacttcgaa aaatcaggtg 240gtgctgacta tgaccaacat ggacccggtg gatacggcca cctattattg cgcgcgtatt 300cagggttgga attatgatgt ttggggccaa ggcaccctgg tgacggttag ctca 354322360DNAHomo sapiens 322caggtgcaat tgcaacagtc tggtccgggc ctggtgaaac cgagccaaac cctgagcctg 60acctgtgcga tttccggaga tagcgtgagc tcttcttctg ctgcttggtc ttggattcgc 120cagtctcctg ggcgtggcct cgagtggctg ggccgtatcg gtcagcgtag caagtggtat 180aacgattatg cggtgagcgt gaaaagccgg attaccatca acccggatac ttcgaaaaac 240cagtttagcc tgcaactgaa cagcgtgacc ccggaagata cggccgtgta ttattgcgcg 300cgttctcgtt ttggttattt tgatgtttgg ggccaaggca ccctggtgac ggttagctca 360323354DNAHomo sapiens 323caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct tctcattatt cttgggtgcg ccaagcccct 120gggaagggtc tcgagtgggt gagcactatc tcttctaatg gtagctatac ctattatgcg 180gatagcgtga aaggccgttt taccatttca cgtgataatt cgaaaaacac cctgtatctg 240caaatgaaca gcctgcgtgc ggaagatacg gccgtgtatt attgcgcgcg ttttgttgct 300cgtcttaatg tttttgatta ttggggccaa ggcaccctgg tgacggttag ctca 354324345DNAHomo sapiens 324caggtgcaat tggttcagtc tggcgcggaa gtgaaaaaac cgggcagcag cgtgaaagtg 60agctgcaaag cctccggagg cactttttct aattatacta tttcttgggt gcgccaagcc 120cctgggcagg gtctcgagtg gatgggcaat atcattccgg cttttggcta tgcgaattac 180gcgcagaagt ttcagggccg ggtgaccatt accgcggatg aaagcaccag caccgcgtat 240atggaactga gcagcctgcg tagcgaagat acggccgtgt attattgcgc gcgtaagttt 300acttttgatg tttggggcca aggcaccctg gtgacggtta gctca 345325354DNAHomo sapiens 325caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct tcttattatt cttgggtgcg ccaagcccct 120gggaagggtc tcgagtgggt gagcaatatc tctggtaatg gtagctctac ctattatgcg 180gatagcgtga aaggccgttt taccatttca cgtgataatt cgaaaaacac cctgtatctg 240caaatgaaca gcctgcgtgc ggaagatacg gccgtgtatt attgcgcgcg tcttattgct 300actcttggta cttttgatta ttggggccaa ggcaccctgg tgacggttag ctca 354326360DNAHomo sapiens 326caggtgcaat tggtggaaag cggcggcggc ctggtgcaac cgggcggcag cctgcgtctg 60agctgcgcgg cctccggatt taccttttct aattatggta ttcattgggt gcgccaagcc 120cctgggaagg gtctcgagtg ggtgagctat atccgttctg gttctagcga tacctattat 180gcggatagcg tgaaaggccg ttttaccatt tcacgtgata attcgaaaaa caccctgtat 240ctgcaaatga acagcctgcg tgcggaagat acggccgtgt attattgcgc gcgtactgct 300cctgctggtc atggtgtttt tgctaattgg ggccaaggca ccctggtgac ggttagctca 360327330DNAHomo sapiens 327gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcgactta ttattgccag cagtattatg attttcctcc tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330328333DNAHomo sapiens 328gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcggtgta ttattgccag cagtattctc atgatccttc tggtaccttt 300ggccagggta cgaaagttga aattaaacgt acg 333329330DNAHomo sapiens

329gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcgactta ttattgccag cagtattatg attttcctcc tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330330330DNAHomo sapiens 330gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcgactta ttattgccag cagtattatg attttcctcc tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330331330DNAHomo sapiens 331gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcggtgta ttattgccag cagttttatt ctaagcctat tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330332333DNAHomo sapiens 332gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcggtgta ttattgccag cagtattctc atgatccttc tggtaccttt 300ggccagggta cgaaagttga aattaaacgt acg 333333333DNAHomo sapiens 333gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcggtgta ttattgccag cagtattctc aggatccttc ttctaccttt 300ggccagggta cgaaagttga aattaaacgt acg 333334330DNAHomo sapiens 334gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gattgtttcc ggttatctgg cttggtacca gcagaaacca 120ggtcaagcac cgcgtctatt aatttatggt gcttcttctc gtgcaactgg ggtcccggcg 180cgttttagcg gctctggatc cggcacggat tttaccctga ccattagcag cctggaacct 240gaagactttg cggtgtatta ttgccagcag tggtctcttc gttctccttt tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330335333DNAHomo sapiens 335gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt tctaattatg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat aatgataatc agcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgctct acttatgatc gtcgtacttt ttctgtgttt 300ggcggcggca cgaagttaac cgtcctaggt cag 333336333DNAHomo sapiens 336gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt tctaattatg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat aatgataatc agcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgctct acttatgatc gtcgtacttt ttctgtgttt 300ggcggcggca cgaagttaac cgtcctaggt cag 333337330DNAHomo sapiens 337gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttcgt tcttctcgtc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggtgcttcta agcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcgactta ttattgccag cagtattatg attttcctcc tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330338336DNAHomo sapiens 338gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt tctaattatg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat aatgataatc agcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgctct tcttgggatc gtgctgatgg ttcttatgtg 300tttggcggcg gcacgaagtt aaccgtccta ggtcag 336339339DNAHomo sapiens 339gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt ggttataatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattctgttt ctaagcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc ggtgcttggg ctcatatgtc tcttggtaag 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339340339DNAHomo sapiens 340gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt ggttataatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattctgttt ctaagcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc gctacttggg atcattctca gatgggtaag 300gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcag 339341339DNAHomo sapiens 341gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt ggttataatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattctgttt ctaagcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc gctacttggg atcattctca gatgggtaag 300gtgtttggcg gcggcacgaa gttaaccgtc ctaggtcag 339342327DNAHomo sapiens 342gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gattgtttcc ggttatctgg cttggtacca gcagaaacca 120ggtcaagcac cgcgtctatt aatttatggt gcttcttctc gtgcaactgg ggtcccggcg 180cgttttagcg gctctggatc cggcacggat tttaccctga ccattagcag cctggaacct 240gaagactttg cgacttatta ttgccagcag tattataatt tttcttttac ctttggccag 300ggtacgaaag ttgaaattaa acgtacg 327343327DNAHomo sapiens 343gatatccaga tgacccagag cccgtctagc ctgagcgcga gcgtgggtga tcgtgtgacc 60attacctgca gagcgagcca gaatattggt tcttatctga attggtacca gcagaaacca 120ggtaaagcac cgaaactatt aatttataat tcttctactt tgcaaagcgg ggtcccgtcc 180cgttttagcg gctctggatc cggcactgat tttaccctga ccattagcag cctgcaacct 240gaagactttg cgacttatta ttgccagcag cgttctaata tgcctattac ctttggccag 300ggtacgaaag ttgaaattaa acgtacg 327344333DNAHomo sapiens 344gatatcgcac tgacccagcc agcttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtacgg gtactagcag cgatgttggt gcttctgata ctgtgacttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tatgctgtta ataagcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc gcttctcgtg attcttcttc tatggtgttt 300ggcggcggca cgaagttaac cgttcttggc cag 333345342DNAHomo sapiens 345gatatcgtga tgacccagag cccactgagc ctgccagtga ctccgggcga gcctgcgagc 60attagctgca gaagcagcca aagcctgctt cattctaatg gctatactta tctgtcttgg 120taccttcaaa aaccaggtca aagcccgcag ctattaattt atcttggttc taagcgtgcc 180agtggggtcc cggatcgttt tagcggctct ggatccggca ccgattttac cctgaaaatt 240agccgtgtgg aagctgaaga cgtgggcgtg tattattgcc agcagtatta tgattcttct 300tctacctttg gccagggtac gaaagttgaa attaaacgta cg 342346336DNAHomo sapiens 346gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt cctaattatg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact tctgattcat ggtaatgcta atcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgccag tcttatgatt tttttactaa ttcttctgtg 300tttggcggcg gcacgaagtt aaccgttctt ggccag 336347330DNAHomo sapiens 347gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctcttact tctaatcagc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat gattcttcta atcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcggttta ttattgccag cagtatggtt cttttcctgc tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330348339DNAHomo sapiens 348gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt ggtaataatt ttgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattatggtg attctcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc cagtcttggg atgctcctat gggtatgtgg 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339349339DNAHomo sapiens 349gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt ggtaataatt ttgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattatggtg attctcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc cagtcttggg atgctcctat gggtatgtgg 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339350336DNAHomo sapiens 350gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt aattttaatt atgtgaattg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattctgttt cttctcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc ggtgcttata ctactgatac tctttctgtg 300tttggcggcg gcacgaagtt aaccgttctt ggccag 336351330DNAHomo sapiens 351gatatcgtgc tgacccagag cccggcgacc ctgagcctgt ctccgggcga acgtgcgacc 60ctgagctgca gagcgagcca gtctgttact tctaattatc tggcttggta ccagcagaaa 120ccaggtcaag caccgcgtct attaatttat ggttcttctt ctcgtgcaac tggggtcccg 180gcgcgtttta gcggctctgg atccggcacg gattttaccc tgaccattag cagcctggaa 240cctgaagact ttgcggttta ttattgccag cagtattctg atattcctgc tacctttggc 300cagggtacga aagttgaaat taaacgtacg 330352327DNAHomo sapiens 352gatatccaga tgacccagag cccgtctagc ctgagcgcga gcgtgggtga tcgtgtgacc 60attacctgca gagcgagcca gtctattaat cgttctctga cttggtacca gcagaaacca 120ggtaaagcac cgaaactatt aatttatgct gcttctaatt tgcaaagcgg ggtcccgtcc 180cgttttagcg gctctggatc cggcactgat tttaccctga ccattagcag cctgcaacct 240gaagactttg cggtttatta ttgccttcag gtttataatc ttcctcttac ctttggccag 300ggtacgaaag ttgaaattaa acgtacg 327353339DNAHomo sapiens 353gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatattggt ggttttaatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tatgatgttt ctaatcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc tctacttatg atgatgatca gcaggatgct 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339354330DNAHomo sapiens 354gatatcgaac tgacccagcc gccttcagtg agcgttgcac caggtcagac cgcgcgtatc 60tcgtgtagcg gcgataatat tcctaatttt tatgttcatt ggtaccagca gaaacccggg 120caggcgccag ttcttgtgat ttatgaggat tctgatcgtc cctcaggcat cccggaacgc 180tttagcggat ccaacagcgg caacaccgcg accctgacca ttagcggcac tcaggcggaa 240gacgaagcgg attattattg ccagtcttat gataagccta ctttttctgg tgtgtttggc 300ggcggcacga agttaaccgt tcttggccag 330355339DNAHomo sapiens 355gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatattggt cgttatcatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa agttatgatt tattctgttt ctaagcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc tctgcttatg atactaataa ttatctttct 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339356324DNAHomo sapiens 356gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt aataattctg tgaattggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat aataatcagc gtccctcagg cgtgccggat 180cgttttagcg gatccaaaag cggcaccagc gcgagccttg cgattacggg cctgcaaagc 240gaagacgaag cggattatta ttgccaggct tatgcttcta atattgtgtt tggcggcggc 300acgaagttaa ccgttcttgg ccag 324357333DNAHomo sapiens 357gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt gattataatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tatcgtgttg ataatcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc cagtcttggg ttggtccttc tactgtgttt 300ggcggcggca cgaagttaac cgttcttggc cag 333358333DNAHomo sapiens 358gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt aataattatg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat tctaattctc agcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgccag tcttatgatc ataattctta tactgtgttt 300ggcggcggca cgaagttaac cgttcttggc cag 333359324DNAHomo sapiens 359gatatcgaac tgacccagcc gccttcagtg agcgttgcac caggtcagac cgcgcgtatc 60tcgtgtagcg gcgataatct tcgttctaag tatgctcatt ggtaccagca gaaacccggg 120caggcgccag ttcttgtgat ttattctaag gataatcgtc cctcaggcat cccggaacgc 180tttagcggat ccaacagcgg caacaccgcg accctgacca ttagcggcac tcaggcggaa 240gacgaagcgg attattattg ctcttcttgg gctcatgatc ataaggtgtt tggcggcggc 300acgaagttaa ccgttcttgg ccag 324360330DNAHomo sapiens 360gatatcgaac tgacccagcc gccttcagtg agcgttgcac caggtcagac cgcgcgtatc 60tcgtgtagcg gcgataatat tggttctaag gttgctactt ggtaccagca gaaacccggg 120caggcgccag ttcttgtgat ttattatgat aatgatcgtc cctcaggcat cccggaacgc 180tttagcggat ccaacagcgg caacaccgcg accctgacca ttagcggcac tcaggcggaa 240gacgaagcgg attattattg ccagtcttat gatggtcaga tgtctacttc tgtgtttggc 300ggcggcacga agttaaccgt tcttggccag 330361339DNAHomo sapiens 361gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt ggttataatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattctgtta ataatcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc tctacttatg atcatacttc ttctggtttt 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339362333DNAHomo sapiens 362gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatattggt cattttaatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tattctgtta tttctcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc gcttctttta cttttccttc tcttgtgttt 300ggcggcggca cgaagttaac cgttcttggc cag 333363336DNAHomo sapiens 363gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt tctcatactg tgaattggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat gatgttaata agcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgcgct tcttgggatt ctgttcaggt ttctcctgtg 300tttggcggcg gcacgaagtt aaccgttctt ggccag 336364333DNAHomo sapiens 364gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt aataatcgtg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat ggtaataata agcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgctct gcttgggatc ttcttgaggt ttatgtgttt 300ggcggcggca cgaagttaac cgttcttggc cag 333365333DNAHomo sapiens 365gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatattggt acttataatc atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tataatgtta ataagcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc tctgcttatg ctccttctgc tgttgtgttt 300ggcggcggca cgaagttaac cgttcttggc cag 333366339DNAHomo sapiens 366gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtacgg gcagcagcag caacattggt attggttatg atgtgaattg gtaccagcag 120ttgcccggga cggcgccgaa acttctgatt tataagaata ctaatcgtcc ctcaggcgtg 180ccggatcgtt ttagcggatc caaaagcggc accagcgcga gccttgcgat tacgggcctg 240caaagcgaag acgaagcgga ttattattgc tcttcttatg attcttttca tcatggtatt 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339367339DNAHomo sapiens 367gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatcttggt ggttattctt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa acttatgatt tataatgtta ataatcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc tcttcttatg attatgtttc ttctgatact 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339368339DNAHomo sapiens 368gatatcgcac tgacccagcc agcttcagtg agcggctcac caggtcagag cattaccatc 60tcgtgtacgg gtactagcag cgatgttggt ggttataatt atgtgtcttg gtaccagcag 120catcccggga aggcgccgaa agttatgatt tatggtgtta ctaagcgtcc ctcaggcgtg 180agcaaccgtt ttagcggatc caaaagcggc aacaccgcga

gcctgaccat tagcggcctg 240caagcggaag acgaagcgga ttattattgc cagacttatg atgagcatgg ttttcatatt 300gtgtttggcg gcggcacgaa gttaaccgtt cttggccag 339369336DNAHomo sapiens 369gatatcgtgc tgacccagcc gccttcagtg agtggcgcac caggtcagcg tgtgaccatc 60tcgtgtagcg gcagcagcag caacattggt aattatcgtg tgtcttggta ccagcagttg 120cccgggacgg cgccgaaact tctgatttat ggtaataata agcgtccctc aggcgtgccg 180gatcgtttta gcggatccaa aagcggcacc agcgcgagcc ttgcgattac gggcctgcaa 240agcgaagacg aagcggatta ttattgctct acttatactg gtcttccttt tactactgtg 300tttggcggcg gcacgaagtt aaccgttctt ggccag 336370327PRTHomo sapiens 370Glu Val Glu Lys Thr Ala Cys Pro Ser Gly Lys Lys Ala Arg Glu Ile 1 5 10 15 Asp Glu Ser Leu Ile Phe Tyr Lys Lys Trp Glu Leu Glu Ala Cys Val 20 25 30 Asp Ala Ala Leu Leu Ala Thr Gln Met Asp Arg Val Asn Ala Ile Pro 35 40 45 Phe Thr Tyr Glu Gln Leu Asp Val Leu Lys His Lys Leu Asp Glu Leu 50 55 60 Tyr Pro Gln Gly Tyr Pro Glu Ser Val Ile Gln His Leu Gly Tyr Leu 65 70 75 80 Phe Leu Lys Met Ser Pro Glu Asp Ile Arg Lys Trp Asn Val Thr Ser 85 90 95 Leu Glu Thr Leu Lys Ala Leu Leu Glu Val Asn Lys Gly His Glu Met 100 105 110 Ser Pro Gln Val Ala Thr Leu Ile Asp Arg Phe Val Lys Gly Arg Gly 115 120 125 Gln Leu Asp Lys Asp Thr Leu Asp Thr Leu Thr Ala Phe Tyr Pro Gly 130 135 140 Tyr Leu Cys Ser Leu Ser Pro Glu Glu Leu Ser Ser Val Pro Pro Ser 145 150 155 160 Ser Ile Trp Ala Val Arg Pro Gln Asp Leu Asp Thr Cys Asp Pro Arg 165 170 175 Gln Leu Asp Val Leu Tyr Pro Lys Ala Arg Leu Ala Phe Gln Asn Met 180 185 190 Asn Gly Ser Glu Tyr Phe Val Lys Ile Gln Ser Phe Leu Gly Gly Ala 195 200 205 Pro Thr Glu Asp Leu Lys Ala Leu Ser Gln Gln Asn Val Ser Met Asp 210 215 220 Leu Ala Thr Phe Met Lys Leu Arg Thr Asp Ala Val Leu Pro Leu Thr 225 230 235 240 Val Ala Glu Val Gln Lys Leu Leu Gly Pro His Val Glu Gly Leu Lys 245 250 255 Ala Glu Glu Arg His Arg Pro Val Arg Asp Trp Ile Leu Arg Gln Arg 260 265 270 Gln Asp Asp Leu Asp Thr Leu Gly Leu Gly Leu Gln Gly Gly Ile Pro 275 280 285 Asn Gly Tyr Leu Val Leu Asp Leu Ser Met Gln Glu Ala Leu Ser Gly 290 295 300 Thr Pro Cys Leu Leu Gly Pro Gly Pro Val Leu Thr Val Leu Ala Leu 305 310 315 320 Leu Leu Ala Ser Thr Leu Ala 325 371340PRTHomo sapiens 371Gly Gly Ser Ser Thr Ile Leu Arg Pro Arg Phe Arg Arg Glu Val Glu 1 5 10 15 Lys Thr Ala Cys Pro Ser Gly Lys Lys Ala Arg Glu Ile Asp Glu Ser 20 25 30 Leu Ile Phe Tyr Lys Lys Trp Glu Leu Glu Ala Cys Val Asp Ala Ala 35 40 45 Leu Leu Ala Thr Gln Met Asp Arg Val Asn Ala Ile Pro Phe Thr Tyr 50 55 60 Glu Gln Leu Asp Val Leu Lys His Lys Leu Asp Glu Leu Tyr Pro Gln 65 70 75 80 Gly Tyr Pro Glu Ser Val Ile Gln His Leu Gly Tyr Leu Phe Leu Lys 85 90 95 Met Ser Pro Glu Asp Ile Arg Lys Trp Asn Val Thr Ser Leu Glu Thr 100 105 110 Leu Lys Ala Leu Leu Glu Val Asn Lys Gly His Glu Met Ser Pro Gln 115 120 125 Val Ala Thr Leu Ile Asp Arg Phe Val Lys Gly Arg Gly Gln Leu Asp 130 135 140 Lys Asp Thr Leu Asp Thr Leu Thr Ala Phe Tyr Pro Gly Tyr Leu Cys 145 150 155 160 Ser Leu Ser Pro Glu Glu Leu Ser Ser Val Pro Pro Ser Ser Ile Trp 165 170 175 Ala Val Arg Pro Gln Asp Leu Asp Thr Cys Asp Pro Arg Gln Leu Asp 180 185 190 Val Leu Tyr Pro Lys Ala Arg Leu Ala Phe Gln Asn Met Asn Gly Ser 195 200 205 Glu Tyr Phe Val Lys Ile Gln Ser Phe Leu Gly Gly Ala Pro Thr Glu 210 215 220 Asp Leu Lys Ala Leu Ser Gln Gln Asn Val Ser Met Asp Leu Ala Thr 225 230 235 240 Phe Met Lys Leu Arg Thr Asp Ala Val Leu Pro Leu Thr Val Ala Glu 245 250 255 Val Gln Lys Leu Leu Gly Pro His Val Glu Gly Leu Lys Ala Glu Glu 260 265 270 Arg His Arg Pro Val Arg Asp Trp Ile Leu Arg Gln Arg Gln Asp Asp 275 280 285 Leu Asp Thr Leu Gly Leu Gly Leu Gln Gly Gly Ile Pro Asn Gly Tyr 290 295 300 Leu Val Leu Asp Leu Ser Val Gln Glu Ala Leu Ser Gly Thr Pro Cys 305 310 315 320 Leu Leu Gly Pro Gly Pro Val Leu Thr Val Leu Ala Leu Leu Leu Ala 325 330 335 Ser Thr Leu Ala 340

Claims

1. An isolated human or humanized antibody, antigen-binding region, or functional fragment thereof comprising an antigen-binding region that is specific for Mesothelin (SEQ ID NO:370), wherein said antibody or functional fragment thereof recognize an epitope of mesothelin that is not masked by another extracellular antigen.

2. The isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1, wherein the another extracellular antigen is cancer antigen 125 (CA125)

3. The isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1 comprising a CDR region depicted in Table 7.

4. The isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1 comprising a heavy chain or light chain amino acid sequence selected from (i) a SEQ ID NO depicted in Table 7; or (ii) a sequence having at least 60 percent sequence identity to a SEQ ID NO depicted in Table 7.

5. The isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1 that is an IgG.

6. The isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1 comprising a SEQ ID NO depicted in Table 7.

7. The isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1 that is a Fab or scFv antibody fragment.

8. An isolated nucleic acid sequence that encodes an isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1.

9. A nucleic acid sequence encoding a variable heavy chain of an isolated antibody or functional fragment thereof comprising (i) a sequence selected from the group consisting of a SEQ ID NO depicted in Table 7 or (ii) a nucleic acid sequence that hybridizes under high stringency conditions to the complementary strand of a SEQ ID NO depicted in Table 7, wherein said antibody or functional fragment thereof is specific for an epitope of Mesothelin.

10. A vector comprising a nucleic acid sequence according to claim 1.

11. An isolated cell comprising a vector according to claim 10.

12. An isolated cell according to claim 11, wherein said isolated cell is bacterial or a mammalian cell.

13. A pharmaceutical composition comprising an isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1 and a pharmaceutically acceptable carrier or excipient therefore.

14. A pharmaceutical composition according to claim 13 adapted for treating an undesired presence of mesothelin.

15. The isolated human or humanized antibody, antigen-binding region, or functional fragment thereof according to claim 1 that is a synthetic human antibody.

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