Anti-hydroxylase Antibodies And Uses Thereof

Abstract

Antibodies, or antigen-binding portions thereof, to aspartyl (asparaginyl) .beta.-hydroxylase are provided. The anti-aspartyl (asparaginyl) .beta.-hydroxylase antibodies, or antigen-binding portions thereof, can modulate activity of aspartyl (asparaginyl) .beta.-hydroxylase.

Description

[0001] This application is a continuation of U.S. application Ser. No. 12/055,108 filed Mar. 25, 2008, which is a continuation of U.S. application Ser. No. 10/989,462 filed Nov. 15, 2004, which claims the benefit of the filing date of U.S. Ser. No. 60/563,514, filed Apr. 19, 2004, and U.S. Ser. No. 60/520,114, filed Nov. 14, 2003. The contents of these prior applications are hereby incorporated by reference in their entirety.

TECHNICAL FIELD

[0003] The present invention relates to antibodies that recognize aspartyl (asparaginyl) .beta.-hydroxylase and to methods of using those antibodies to, for example, detect aspartyl (asparaginyl) .beta.-hydroxylase and/or modulate its activity.

BACKGROUND

[0004] Aspartyl(asparaginyl) .beta.-hydroxylase (AAH) catalyzes post-translational hydroxylation of .beta. carbons of specific aspartate and asparagine residues in epidermal growth factor-like domains of numerous proteins, including extracellular matrix proteins, low-density lipoprotein (LDL) receptor, Notch homologs, and Notch ligand homologs (Jia et al., Proc. Natl. Acad. Sci. USA 91(15):7227-7231, 1994; Jia et al., J. Biol. Chem. 267(20):14322-14327, 1992; Gronke et al., Proc Natl Acad Sci USA 86(10):3609-13, 1989). This transmembrane enzyme is a member of the .alpha.-ketoglutarate-dependent dioxygenase family of prolyl and lysyl hydroxylases. Overexpression of human AAH (HAAH) has been detected in a number of human cancers, including hepatocellular carcinomas, cholangiocarcinomas, and, neuroectodermal tumors (Lavaissiere et al., J. Clin. Investig. 98:1313-1323, 1996; Sepe et al., Lab. Investig. 82(7):81-891, 2002). The finding that AAH is overexpressed in numerous tumors, and that forced expression increases cell motility and survival indicates that AAH may contribute to malignant transformation in vivo (Sepe et al., Lab. Investig. 82(7):881-891, 2002).

SUMMARY

[0005] This invention is based, in part, on our discovery of human single-chain antibodies that bind aspartyl (asparaginyl) .beta.-hydroxylase (AAH). Accordingly, the invention features antibodies, including complete, multimeric antibodies (e.g., human tetrameric antibodies of the G class (an IgG)) and fragments or other variants thereof that specifically bind an AAH protein. These fragments and variants include single-chain anti-AAH antibodies, fragments or portions of multimeric (e.g., tetrameric) anti-AAH antibodies and other variants that specifically bind an AAH protein. Any of the anti-AAH antibodies or fragments or variants thereof may have been mutagenized by, for example, an affinity maturation process.

[0006] The compositions of the invention include anti-AAH antibodies (e.g., a human monoclonal antibody), fragments or other variants thereof, pharmaceutical compositions containing them, and kits containing them. The antibodies, fragments or other variants thereof may, but do not necessarily, inhibit one or more of AAH's biological activities (e.g., hydroxylation of suitable substrates, whether naturally occurring or non-naturally occurring, in vitro, in cell or tissue culture, or in vivo) and/or one or more of the cellular events mediated by AAH (e.g., cellular growth or proliferation, or cellular motility). As undesirable cellular proliferation and motility occur in connection with pathological conditions in which AAH is overactive or overexpressed (such as cancer), the compositions of the invention can be used to identify patients having such a condition, to assess their prognosis, and/or to treat a patient having, or at risk for developing, such a condition (e.g., cancers or other disorders associated with elevated AAH expression or activity).

[0007] More specifically, the invention features methods of identifying a patient amenable to treatment (e.g., a patient having a cell (or cells) in which AAH is overactive or overexpressed) and therapeutic or prophylactic methods of treating such a patient by administering an effective amount of an anti-AAH antibody (or a fragment or other variant thereof) to the patient (e.g. a human monoclonal antibody or a human single-chain antibody (scFv) that specifically binds HAAH). Also provided are nucleic acids that can be used to express the antibodies of the invention; vectors that include those nucleic acids; cells that contain those nucleic acids or vectors; methods of formulating pharmaceutically acceptable compositions that include an anti-AAH antibody and/or a fragment or other variant thereof; methods of identifying and/or assessing the properties of anti-AAH antibodies (e.g., anti-HAAH antibodies) and/or fragments or other variants thereof; and methods of affinity-maturing anti-AAH antibodies (e.g. human anti-HAAH antibodies) and/or fragments or other variants thereof.

[0008] For ease of reading, we do not repeat the phrase "and/or fragments or other variants thereof" following every occurrence of "antibody" or "antibodies." It is to be understood that wherever an anti-AAH antibody can be used, a fragment or other variant thereof that specifically binds an AAH protein to any useful degree can also be used. We may refer to AAH as "an AAH protein." Unless a contrary meaning is clear, we use the terms "protein," "peptide" and "polypeptide" interchangeably to refer to chains of two or more amino acid residues. Similarly, wherever a nucleic acid encoding an anti-AAH antibody can be used, a nucleic acid encoding a functional fragment or other variant of the anti-AAH antibody can be used; wherever a cell comprising an anti-AAH antibody can be used, one can use a cell comprising a fragment or other variant of an anti-AAH antibody; and so forth. A fragment of an anti-AAH antibody may also be referred to as an "antigen-binding portion" of an anti-AAH antibody, as the fragment may be a portion of an anti-AAH antibody that specifically binds an AAH antigen. While fragments and variants are described further below, we note here that they include Fab, Fab' and F(ab')2 fragments as well as scFvs (e.g., a human scFv that specifically binds HAAH).

[0009] The anti-AAH antibodies or fragments or other variants thereof can consist of, or can include, the amino acid sequences in the Tables of FIGS. 34-38. Alternatively, the anti-AAH antibodies can consist of, or can include, sequences that exhibit a certain degree of identity to the amino acid sequences in the Tables of FIGS. 34-38. For example, the antibodies can include a variable region of the heavy chain (VH) that is at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100% identical) to one of the VH sequences shown in FIGS. 34 and 35. Alternatively, the antibodies can be scFv that are at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to one of the scFvs shown in FIG. 36. Alternatively, the antibodies or fragments or other variants thereof can include complementarity determining regions (CDRs) that are at least 40% identical (e.g., at least 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 98%) identical to the CDRs shown in FIGS. 37 and 38. Other fragments and variants are described further below.

[0010] The antibody can include a variable light chain (VL or VLC) including a first CDR including (or consisting of) the amino acid sequence Ser-Gln-Ser/Asn-Val-Ser-Ser/His-(Xaa)-Tyr/His-Leu-Ala (SEQ ID NO:320); a second CDR including (or consisting of) the amino acid sequence Asp-Val-Ala-Asn-Xaa-Ala-Ala (SEQ ID NO:321); and a third CDR including (or consisting of) the amino acid sequence Gln-Gln-Arg-Ser-Gln-Trp-Pro-Gln (SEQ ID NO:322). Unless indicated otherwise, in these sequences and those that follow, "Xaa" is any amino acid residue or no amino acid residue. Alternatively, or in addition, the antibody can include a VHC including a first CDR including (or consisting of) the amino acid sequence Tyr/His-Ala-Met-His/Gly (SEQ ID NO:323) and a second CDR including (or consisting of) the amino acid sequence Tyr-Ala-Xaa-Ser-Val-Lys-Gly/Ser (SEQ ID NO:324). In other embodiments, the antibody can include a VLC including a CDR (e.g., a first CDR) including (or consisting of) the amino acid sequence Ser-Gly-Ser-Ser-Ser-Asn-Ile-Gly/Glu-Ser-Asn-His/Tyr-Val-His/Tyr (SEQ ID NO:325). Alternatively, or in addition, the antibody can include a VHC including a CDR (e.g., a first CDR) including (or consisting of) the amino acid sequence Ser/Gly-Asp/Asn-Ser/Gly-Ala-Ala-Trp-Ser/Asn (SEQ ID NO:326) and a second CDR including (or consisting of) the amino acid sequence Arg-Ile/Thr-Tyr/His-Tyr/His-Gly/Arg-Xaa-Lys/Arg-Trp/Arg-Tyr/Arg-- Asn-Asp/Gly-Tyr/His-Ala-Val/Ala-Pro/Ser-Val/Ala-Lys-Ser (SEQ ID NO:327). In other embodiments, the antibody can include a VLC including a CDR (e.g., a second CDR) including (or consisting of) the amino acid sequence Asp-Val-Xaa-Xaa-Arg-Pro-Ser (SEQ ID NO:328). Alternatively, the antibody can include a CDR (e.g., a second CDR) including (or consisting of) the amino acid sequence Leu-Phe/Leu-Ile/Val-His/Tyr-Lys/Arg-Xaa-Asn-Gln-Arg-Pro-Ser (SEQ ID NO:329) and, optionally, a CDR (e.g., a third CDR) including (or consisting of) the amino acid sequence Ala-Trp-Asp-Asp-Ser (SEQ ID NO:330). For example, the third CDR can consist of the sequence Ala-Ala-Trp-Asp-Asp-Ser-Leu-Arg-Gly-Tyr-Val (SEQ ID NO:51). The antibody can also include a VHC including a CDR (e.g., a third CDR) including (or consisting of) the amino acid sequence Ser-Ser-Ser-Trp-Val-Val-Xaa-Phe-Asp/Gly (SEQ ID NO:331) (e.g., Thr-Gly-Tyr-Ser-Ser-Ser-Trp-Val-Val-Asn-Phe-Asp-Tyr (SEQ ID NO:96).

[0011] Any of the antibodies of the invention, regardless of their sequence, can be monoclonal (e.g., monospecific) or polyclonal antibodies; any of the antibodies can be human or humanized; any of the antibodies can be affinity matured; and any of the antibodies can be isolated (e.g., purified to some degree from an animal or cells in which they are produced). Human antibodies include antibodies that have variable and constant regions from human germline immunoglobulin sequences. Such antibodies may have all, or a portion of, a human immunoglobulin heavy chain and all, or a portion of, a human immunoglobulin light chain. This is not to say that the antibodies of the invention (human or non-human) must contain naturally occurring sequences. The antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin nucleic acid sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro). Mutated or affinity matured antibodies are described further below.

[0012] While the antibodies are not limited to those that bind an AAH in a particular manner, the antibodies may bind the catalytic domain of an AAH (e.g., a catalytic domain of HAAH) or they may bind AAH in such a way as to alter the conformation of the catalytic domain or otherwise render it less active. The extent to which the activity of the bound AAH antigen is inhibited can vary. Useful antibodies (or fragments or other variants thereof) within the scope of the present invention include those that inhibit an activity of an AAH (e.g., the catalytic activity of HAAH) to a clinically beneficial degree upon administration to a patient or to an extent that they are useful in in vitro assays. For example, the antibodies can inhibit an enzymatic activity of AAH or HAAH by more than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% (e.g., about 95%, 98%, or 99%). The inhibition can be assessed relative to a control or a reference sample or standard. Alternatively, an anti-AAH antibody can specifically bind an AAH protein without inhibiting the activity of the AAH protein. Such antibodies are useflI in detecting AAH and can be used to identify a patient having cells in which AAH is overexpressed. The identification process can take place, for example, prior to administering an anti-AAH antibody to the patient that inhibits a biological activity of (e.g., the catalytic activity of) AAH.

[0013] The antibodies of the invention can be further characterized or assessed in terms of their ability to inhibit tumor cell growth or tumor cell motility in cell culture and/or to inhibit tumor cell growth or metastasis in vivo. Affinity can also be measured. For example, an antibody of the invention can have an affinity for an AAH protein (e.g., HAAH) that is equal to or less than about 1 .mu.M. The antibodies of the invention can also be assessed for competition with other antibodies. For example, an antibody of the invention (e.g. a human or humanized antibody) may compete with an anti-AAH murine antibody (e.g., the monoclonal antibody FB50 or the monoclonal antibody 15C7) for binding to an epitope bound by the murine antibody. FB50 and 15C7 are explicitly excluded from the scope of the present invention. An antibody of the invention may also be assessed for its ability to compete with the clone 11 antibody (described herein (see FIG. 34)) for binding to an epitope bound by the clone 11 antibody. Antibodies that effectively compete with the clone 11 antibody are within the scope of the present invention.

[0014] In view of the foregoing, we may use the terms "specific binding" or "specifically binds to" refer to the ability of an antibody to: (1) bind to an AAH with a useful affinity (e.g. an affinity of at least 1.times.10.sup.6 M.sup.-1, or (2) bind to the AAH with an affinity that is greater than (e.g., at least two-fold greater than) its affinity for a nonspecific antigen, or (3) bind to the AAH with an affinity sufficient to produce a clinically desirable outcome (e.g., an improvement in a sign or symptom of a subject in need of treatment (e.g., a subject who has cancer or other unwanted cellular proliferation)).

[0015] The antibodies can include one or more Fc domains (e.g., an Fc domain of the gamma isotype (e.g., IgG.sub.1)). We use the term "isotype" in its conventional sense to refer to the antibody class (e.g., IgM, IgA, IgE, IgD, or IgG1) that is encoded by heavy chain constant region genes. The antibodies of the invention can be of any isotype. Alternatively, or in addition, the antibodies can include a label (e.g., a polypeptide that serves as a marker or reporter sequence or that facilitates purification of the antibody sequence to which it is attached). Suitable labels include a FLAG tag, a histadine tag, or an enzymatically active or fluorescent protein. Alternatively, or in addition, the antibodies can include a toxin.

[0016] In other aspects, the invention features isolated nucleic acid molecules that include a sequence encoding an antibody of the invention (e.g., an anti-AAH scFv); expression vectors (e.g., plasmids) including a nucleic acid sequence encoding an antibody of the invention; and host cells including one or more types of those nucleic acid molecules or expression vectors (e.g., prokaryotic cells or eukaryotic cells such as a yeast or mammalian cells (e.g., Chinese hamster ovary (CHO) cells or tumor cells (e.g., myeloma cells))). Anti-AAH scFvs can be readily converted to multimeric anti-AAH antibodies (e.g., IgGs). More specifically, the invention features antibodies expressed on the surfaces of cells (e.g., displayed on yeast cells). We may use the term "recombinant" to refer to an antibody that is prepared by recombinant means (e.g., an antibody that is expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant, combinatorial antibody library, antibodies isolated from an animal (e.g., a mouse) that is transgenic for a human immunoglobulin gene (or genes), or antibodies prepared by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences). Recombinant antibodies include humanized or CDR grafted antibodies; chimeric antibodies; and antibodies generated in vitro (e.g., by phage display). The antibodies may include constant regions derived from human germline immunoglobulin sequences.

[0017] The invention also features kits containing one or more of the compositions listed above and instructions (regardless of form; whether printed, audio- or visual) for use. For example, the kits of the invention can include an anti-AAH antibody (in a lyophilized or concentrated form or suspended in a physiologically acceptable diluent at a concentration suitable for use (e.g., at a concentration suitable for performing a diagnostic assay or administration to a patient)). The kits can also include nucleic acids, vectors, and/or host cells as described herein. Optionally, any of the kits of the invention can include paraphernalia for administering an anti-AAH antibody (e.g., needles, syringes, alcohol swabs, and bandages) or for using it in a diagnostic assay (e.g., reagents useful as controls).

[0018] The methods of the invention include methods of evaluating AAH expression in a cell. The cell can be a human or non-human cell and/or a cell in which AAH is overactive or overexpressed; the cell can be a cell in vivo or one maintained in tissue culture; the cell can be cancerous (e.g., a tumor cell); and the cell can be obtained from essentially any tissue type (e.g., a cell of the lung, liver, colon, pancreas, prostate, ovary, bile duct, brain, or breast). The cell may or may not be intact (e.g., a tissue homogenate can be used, as can proteins that have been purified from the cell). The methods can include providing an anti-AAH antibody (any of those described herein can be used); contacting the cell (or tissue homogenate or protein purified from the cell) with the antibody under conditions, and for a time, sufficient to allow the antibody to bind AAH expressed by the cell; and detecting the antibody or antigen-binding portion thereof (by, e.g., Western blot analysis, immunohistochemistry, or other antibody-based detection method). The detection step may provide a qualitative or quantitative assessment of AAH expression, and the result can be compared with that obtained from a control sample or with a reference sample or standard. Such methods can be carried out effectively with antibodies that specifically bind AAH, but have no impact on AAH's activity. The methods may be performed as part of an effort to diagnose patients amenable to treatment with the AAH-inhibitory antibodies of the invention.

[0019] The methods of the invention also include methods of modulating (e.g., inhibiting) the activity of an AAH protein in a cell. As with the evaluative methods described above, the cell can be a human or non-human cell and/or a cell in which AAH is overactive or overexpressed; the cell can be a cell in vivo or one maintained in tissue culture; the cell can be cancerous (e.g., a tumor cell); and the cell can be obtained from essentially any tissue type (e.g., a cell of the lung, liver, colon, pancreas, prostate, ovary, bile duct, brain, or breast). The methods can include providing a cell and exposing that cell to an antibody (or fragment or other variant) described herein, for a time, and under conditions sufficient to modulate (e.g., inhibit) AAH activity in the cell. When the cell is exposed to an anti-AAH antibody in vivo, the methods can be described as methods for treating a patient who has a disorder (e.g., a cancer or other disorder of unwanted cellular proliferation) or who is a risk for developing such a disorder (e.g., a patient in which AAH is overactive or overexpressed but who does not yet have a detectable tumor or other sign of cancer). The methods can include administering an antibody (or fragment or other variant thereof) described herein to the patient in an amount and for a time sufficient for the antibody to inhibit proliferation or metastasis of a cancerous cell in the patient. While methods of treatment are described further below, we note that a composition including an anti-AAH antibody can be administered locally (e.g., to the site of a tumor or to tissue remaining after a tumor has been surgically removed) or systemically (e.g., by intravenous injection). The patient can receive a single type of anti-AAH antibody or a combination of antibodies, and the antibody (or antibodies) can be administered in combination with a second agent (e.g. a second chemotherapeutic agent, an analgesic, or anti-emetic).

[0020] Terms relating to treatment refer to the application or administration of a composition of the invention to a patient or to a cell provided from a patient. The composition can be an anti-AAH antibody, a nucleic acid molecule or expression vector encoding same, or a host cell expressing same (any of which can be combined with a physiologically acceptable diluent). The composition can be administered ex vivo to cells isolated from (e.g., removed from) a subject, preferably from the patient in need of treatment. Upon conclusion of the treatment, the cells can be returned to the patient. Moreover, the treatment methods can be prophylactic. For example, they can be applied to a patient who is at risk for developing cancer (there are well established indicators of risk (e.g., levels of cancer-associated antigens, such as PSA and, as noted, AAH per se)). The treatment can be one that cures or heals the patient, but the invention is not so limited. The methods of the invention may also alleviate, relieve, alter, ameliorate, palliate, or improve a sign or symptom of the cancer or the patient's predisposition toward the cancer.

[0021] A "therapeutically effective amount" of an anti-AAH antibody is an amount of an anti-AAH antibody effective to treat a sign or symptom of a disorder (e.g., a cancer, e.g., a tumor or other neoplasm or dysplastic syndrome). A "prophylactically effective amount" of an anti-AAH antibody is an amount of an anti-AAH antibody effective in delaying the occurrence of the onset or recurrence of a disorder (e.g., a cancer), or reducing the severity of a sign or symptom thereof.

[0022] While patients amenable to treatment are described further below, we note here that the patient can have any proliferative disorder associated with overactive or overexpressed AAH. For example, the patient can have a tumor within the lung, liver, colon, pancreas, prostate, ovary, bile duct, brain, or breast containing AAH-positive cells.

[0023] In another aspect, the invention features methods of identifying an antibody that specifically binds to an AAH. The methods can include, for example: (a) providing a library of antibodies (e.g., human antibodies, which may be scFvs); (b) contacting members of the library with AAH proteins or fragments thereof, under conditions that allow the antibodies to bind the polypeptides; and (c) selecting an antibody that binds to the AAH protein (or AAH fragment). The method can further include affinity-maturing the selected antibody. Maturation can be achieved with, for example, methods such as error-prone PCR, or methods in which a nucleic acid encoding the antibody is subjected to recombination with nucleic acid(s) of a library, e.g., using CDR shuffling or chain shuffling techniques. The invention also features methods for making a human monoclonal antibody that specifically binds to an AAH. The methods can include the steps of: identifying an antibody that specifically binds to an AAH protein; expressing a nucleic acid sequence encoding the antibody in a cell; and isolating the expressed antibody from the cell. We may refer to these methods as production methods, and anti-AAH antibodies made by these methods are within the scope of the present invention.

[0024] The antibodies of the invention may be advantageous for various reasons. For example, when less than full-length antibodies are used, the antibody fragments or variants may penetrate tumors more readily. When human or humanized antibodies are administered to human patients, they are unlikely to stimulate an undesirable immune response as potent as that triggered by non-human proteins. The antibodies of the invention are also unlikely to generate other undesirable side effects because HAAH expression is very low or undetectable in non-cancerous tissues, and antibodies directed to HAAH may be less toxic to non-cancerous tissue than less specific treatments. To our knowledge, we have the first human antibodies that bind to HAAH.

[0025] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0026] FIG. 1 is a schematic diagram depicting the fusion proteins displayed on yeast cells for performing screening and binding studies with the yeast surface display technique. Also depicted are various epitope tags and an antigen that can be employed for detection. The fluorescent antigen used in the experiments described herein was a recombinant HAAH protein corresponding to full-length or catalytic domain only, as indicated.

[0027] FIG. 2 is a set of graphs depicting the fluorescence detected by flow cytometry in experiments in which three unique scFv clones expressed on yeast cells were incubated with fluorescent HAAH.

[0028] FIG. 3 is a set of graphs depicting the fluorescence detected by flow cytometry in experiments in which eight unique scFv clones expressed on yeast cells were incubated with a fluorescently labeled fragment of HAAH containing the catalytic domain.

[0029] FIG. 4 is a set of graphs depicting affinity measurements for two anti-HAAH scFv antibody clones.

[0030] FIG. 5 is a set of graphs depicting the fluorescence detected by flow cytometry in experiments in which one scFv clone expressed on yeast cells was incubated with a fluorescently labeled catalytic domain of HAAH or conjugated full-length HAAH. The conjugated full length HAAH proteins were detected using FB50 IgG or 15C7 IgG, as indicated.

[0031] FIG. 6 is a bar graph depicting binding of soluble scFv fragments of 6 unique clones to the catalytic domain of HAAH.

[0032] FIG. 7 is a bar graph depicting binding of soluble scFv fragments of 6 unique clones to full length HAAH.

[0033] FIG. 8 is a set of graphs depicting the fluorescence detected by flow cytometry in experiments in which scFv fragments of three unique clones bound to H640 tumor cells.

[0034] FIG. 9 is a bar graph depicting the percentage of motile H460 tumor cells in the presence of scFv fragments of 5 unique clones, 15C7 (mouse IgG), and in the absence of antibody (first bar).

[0035] FIG. 10 is a graph depicting the fluorescence detected by flow cytometry in an experiment in which affinity matured scFv fragments derived from clone 11 bound to the catalytic domain of HAAH.

[0036] FIG. 11 is a depiction of the amino acid sequence of an affinity-matured clone 11 scFv fragment. Amino acid residues that are changed relative to clone 11 are bold and underlined.

[0037] FIG. 12 is a set of dot-plot graphs depicting two types of fluorescence detected by flow cytometry in experiments in which the original clone 11 scFv fragments, a first round of mutant clone 11 scFv fragments, and a clone derived from the first round of mutants were analyzed. Fluorescence intensity depicted on the X-axis corresponds to the level of scFv fragments displayed on yeast cells. Fluorescence intensity on the Y-axis corresponds to the level of binding to HAAH of the cells.

[0038] FIG. 13 is a set of dot-plot graphs depicting two types of fluorescence detected by flow cytometry in experiments in which the mutant clone 11 scFv fragments were analyzed. Fluorescence intensity depicted on the X-axis corresponds to the level of scFv fragments displayed on yeast cells. Fluorescence intensity on the Y-axis corresponds to the level of binding to HAAH of the cells.

[0039] FIG. 14 is a graph depicting the fluorescence detected by flow cytometry in an experiment in which affinity matured scFv fragments derived from clone 13 bound to the catalytic domain of HAAH.

[0040] FIG. 15 is a depiction of the amino acid sequence of an affinity-matured clone 13 scFv fragment, 13m1. Amino acid residues that are changed relative to clone 13 are bold and underlined.

[0041] FIG. 16 is a set of dot-plot graphs depicting two types of fluorescence detected by flow cytometry in experiments in which the original clone 13 scFv fragments, a first round mutant clone 13 scFv, and a second round mutant clone 13 scFv, were analyzed. Fluorescence intensity depicted on the X-axis corresponds to the level of scFv displayed on yeast cells. Fluorescence intensity on the Y-axis corresponds to the level of binding to HAAH of the cells.

[0042] FIG. 17 is a schematic depiction of a DNA plasmid with restriction enzyme sites for chain shuffling.

[0043] FIG. 18A is a chart listing the VH and VL regions in a wild type clone and five clones generated by chain shuffling from the wild type clone.

[0044] FIG. 18B is a dot-plot graph depicting two types of fluorescence detected by flow cytometry in experiments in which the pool of mutant clones derived from wild type clone 11 by chain shuffling were analyzed. Fluorescence intensity depicted on the X-axis corresponds to the level of scFv fragments displayed on yeast cells. Fluorescence intensity on the Y-axis corresponds to the level of binding to HAAH of the cells.

[0045] FIGS. 19A-K are dot-plot graphs depicting two types of fluorescence detected by flow cytometry in experiments in which the clones produced by chain shuffling were analyzed (FIG. 19A, clone LLm1; FIG. 19B, clone LLm3; FIG. 19C, clone LLm5; FIG. 19D, clone LLm6; FIG. 19E, clone LLm7; FIG. 19F, clone LLm8; FIG. 19G, clone LLm9; FIG. 19H, clone LLm11; FIG. 19I, clone LLm14; FIG. 19J, clone LLm15(3); FIG. 19K, clone LLm20(8)). Fluorescence intensity depicted on the X-axis corresponds to the level of scFv fragments displayed on yeast cells. Fluorescence intensity on the Y-axis corresponds to the level of binding to HAAH of the cells.

[0046] FIG. 20 is a depiction of the amino acid sequences of CDR1, CDR2, and CDR3 regions of a wild-type scFv clone and eleven clones derived by chain shuffling.

[0047] FIG. 21 is a schematic depiction of an acceptor DNA plasmid encoding a wild type clone 11 scFv and a human library insert sequence to be recombined with the plasmid for CDR shuffling.

[0048] FIGS. 22A-D are dot-plot graphs depicting two types of fluorescence detected by flow cytometry in experiments in which the clones produced by chain shuffling were analyzed (FIG. 22A, CM1; FIG. 22B, CM2; FIG. 22C, CM3; FIG. 22D, CM4). Fluorescence intensity depicted on the X-axis corresponds to the level of scFv fragments displayed on yeast cells. Fluorescence intensity on the Y-axis corresponds to the level of binding to HAAH of the cells.

[0049] FIG. 23 is a depiction of the amino acid sequences of CDR1, CDR2, and CDR3 regions of a wild-type scFv clone, and four clones derived by CDR shuffling.

[0050] FIG. 24 is a graph depicting the concentration of selected anti-HAAH scFv fragments vs. anti-HAAH binding activity (as measured by fluorescence).

[0051] FIG. 25 is a schematic diagram depicting the conversion of an scFv fragment to a full length IgG antibody.

[0052] FIGS. 26A-B are graphs depicting the binding of wild-type anti-HAAH IgG antibodies to HAAH as determined by ELISA. The concentration used for each antibody is indicated below the graphs.

[0053] FIG. 27 is a graph depicting the binding of wild-type anti-HAAH IgG antibodies to H460 tumor cells as determined by FACS. The concentrations of antibody used are indicated on the graph.

[0054] FIG. 28 is a graph depicting the binding of selected anti-HAAH IgG mutant antibodies to H460 tumor cells as determined by FACS. The concentrations of antibody used are indicated on the graph.

[0055] FIG. 29A is a graph depicting the concentration of 6-22 IgG antibody vs. binding to H460 cells (as measured by fluorescence).

[0056] FIG. 29B is a graph depicting the concentration of 6-22 IgG antibody vs. binding to FOCUS cells (as measured by fluorescence).

[0057] FIG. 30A is a graph depicting the concentration of CDRm4 IgG antibody vs. binding to H460 cells (as measured by fluorescence).

[0058] FIG. 30B is a graph depicting the concentration of CDRm4 IgG antibody vs. binding to FOCUS cells (as measured by fluorescence).

[0059] FIG. 31 is a graph depicting the binding of scFv 6-22 to H460 cells in the presence or absence of competition from 6-22 IgG antibody.

[0060] FIG. 32A is a graph depicting the binding of HAAH to yeast expressing different scFv fragments in the presence or absence of competition by CDRm4 IgG antibody.

[0061] FIG. 32B is a graph depicting the binding of HAAH to yeast expressing different scFv fragments in the presence or absence of competition by LLm11 IgG antibody.

[0062] FIG. 33A is a graph depicting the binding of HAAH to yeast displaying scFv CDRm4 and the second generation mutant scFv C4m18. Dissociation constants are indicated on the graph.

[0063] FIG. 33B is a graph depicting the display level of first and second generation scFv mutants on yeast.

[0064] FIG. 34 is a Table containing amino acid sequences that can be used as, or that can be included in, anti-AAH antibodies or fragments or other variants thereof.

[0065] FIG. 35 is a Table containing amino acid sequences that can be used as, or that can be included in, anti-AAH antibodies or fragments or other variants thereof (e.g., anti-HAAH affinity-matured, mutagenized antibody regions).

[0066] FIG. 36 is a Table containing amino acid sequences that can be used as, or that can be included in, anti-AAH antibodies or fragments or other variants thereof (e.g., anti-HAAH scFv antibodies).

[0067] FIG. 37 is a Table containing amino acid sequences (anti-AAH antibody CDRs) that can be included in anti-AAH antibodies or fragments or other variants thereof.

[0068] FIG. 38 is a Table containing amino acid sequences (affinity-matured, mutagenized anti-HAAH antibody CDRs) that can be included in anti-AAH antibodies or fragments or other variants thereof.

DETAILED DESCRIPTION

[0069] Aspartyl(asparaginyl) .beta.-hydroxylase (AAH) is a highly conserved alpha-ketoglutarate-dependent dioxygenase that catalyzes post-translational hydroxylation of .beta. carbons of specific aspartate and asparagine residues in epidermal growth factor-like domains of numerous proteins, including extracellular matrix proteins, low-density lipoprotein (LDL) receptor, Notch homologs, and Notch ligand homologs (Jia et al, Proc. Natl. Acad. Sci. USA 91(15):7227-7231, 1994; Jia et al., J. Biol. Chem. 267(20):14322-14327, 1992; Gronke et al., Proc Natl Acad Sci USA 86(10):3609-13, 1989). Overexpression of human AAH (HAAH) has been detected in a number of human cancers, including hepatocellular carcinomas, cholangiocarcinomas, and, neuroectodermal tumors (Lavaissiere et al, J. Clin. Investig. 98:1313-1323, 1996; Sepe et al., Lab. Investig. 82(7):81-891, 2002). The finding that AAH is overexpressed in numerous tumors, and that forced expression increases cell motility and survival indicates that AAH may contribute to malignant transformation in vivo (Sepe et al., Lab. Investig. 82(7):881-891, 2002). Inventions relating to the use of HAAH for diagnosis and treatment of cancer have been described (Radosevich, U.S. Pat. No. 6,166,176; Radosevich, U.S. Pat. No. 6,727,080; Wands et al, U.S. Pat. No. 6,783,758; Wands et al., U.S. Pat. No. 6,797,696; Wands et al, U.S. Pat. No. 6,812,206; Wands et al, U.S. Pat. No. 6,815,415).

[0070] The present invention relates to antibodies and antigen binding portions thereof having binding specificity for AAH or a portion of AAH. In particular, the invention relates to human monoclonal antibodies that specifically bind to AAH. In one embodiment, the antibodies or antigen binding portions thereof have specificity for human AAH (HAAH). Antibodies that inhibit one or more functions characteristic of a AAH are within the scope of the present invention, whether that function is an enzymatic activity (e.g., hydroxylase activity) or a function manifested on the cellular level (e.g., facilitating motility of a tumor cell). Thus, for example, an anti-AAH antibody can be one that inhibits motility of a tumor cell. Alternatively, or in addition, an antibody of the invention may inhibit (reduce or prevent) the interaction of AAH with a natural ligand, such as a protein containing an EGF-like domain (e.g., an extracellular matrix protein). Human monoclonal antibodies directed against (e.g., antibodies that specifically bind) AAH can inhibit functions mediated by AAH, including modulation of substrate activity by hydroxylation. Preferably, the antibodies and antigen binding portions thereof can bind AAH with an affinity of greater than 1.times.10.sup.6 M.sup.-1.

[0071] The HAAH amino acid sequence is found in GenBank.RTM. under accession number I38423 (GI:7433245). One of ordinary skill in the art can readily retrieve the sequence from GenBank.RTM. or another source. The transmembrane domain of HAAH is designated as being between amino acids 341-374 of the GenBank.RTM. sequence. The extracellular (or luminal) portion of the molecule corresponds to the C-terminal end. Anti-AAH antibodies, whether full-length or not, will interact with (e.g., bind to) AAH or a fragment of that protein (e.g. an anti-HAAH antibody will bind to HAAH). The antibody may bind to an epitope of AAH (e.g., a conformational or a linear epitope) or to a fragment of the full-length AAH protein. Conformational epitopes are typically lost when exposed to a denaturing solvent.

[0072] Antibodies: Antibodies of the invention can assume various configurations. For example, the antibody can be a tetramer (e.g. an antibody having two heavy chains and two light chains) or a single-chain antibody. Accordingly, the antibodies of the invention include proteins that may have one or two heavy (H) chain variable regions, and one or two light chain variable regions. The VHC and VLC regions can be further subdivided into regions of hypervariability, termed "complementarity determining regions" (CDRs), interspersed with regions that are more conserved, termed "framework regions" (FRs). The extent of the FRs and CDRs has been defined (see, Kabat, E. A., et al. Sequences of Proteins of Immunological Interest Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242, 1991, and Chothia, et al., J. Mol. Biol. 196:901-917, 1987, which are incorporated herein by reference). Where an antibody of the invention includes one or more VHCs and/or one or more VLCs, each VHC and VLC can be composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0073] The VHC or VLC chain of an antibody of the invention can further include all or part of a heavy or light chain constant region. In one embodiment, the antibody is a tetramer of two heavy immunoglobulin chains and two light immunoglobulin chains, wherein the heavy and light immunoglobulin chains are inter-connected by, e.g., disulfide bonds. The heavy chain constant region includes three domains: CH1, CH2 and CH3. The light chain constant region is comprised of one domain: CL. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies typically mediate the binding of the antibody to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. The term "antibody" includes intact immunoglobulins of types IgA, IgG, IgE, IgD, IgM (as well as subtypes thereof (e.g., IgG.sub.1, IgG.sub.2, IgG.sub.3, and IgG.sub.4)), wherein the light chains of the immunoglobulin may be of types kappa or lambda.

[0074] Antibodies may also be referred to as "immunoglobulins" (proteins consisting of one or more polypeptides substantially encoded by immunoglobulin genes, the anti-AAH antibodies of the invention may also be referred to as anti-AAH immunoglobulins, and may contain sequences encoded by one or more of the human immunoglobulin genes). The recognized human immunoglobulin genes include the kappa, lambda, alpha (IgA.sub.1 and IgA.sub.2), gamma (IgG.sub.1, IgG.sub.2, IgG.sub.3, IgG.sub.4), delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Full-length immunoglobulin "light chains" (about 25 kDa and 214 amino acids) are encoded by a variable region gene at the NH.sub.2-terminus (about 110 amino acids) and a kappa or lambda constant region gene at the COOH-terminus. Full-length immunoglobulin heavy chains (about 50 kDa and 446 amino acids), are similarly encoded by a variable region gene (about 116 amino acids) and one of the other aforementioned constant region genes, e.g., gamma (encoding about 330 amino acids). The antibodies or immunoglobulins of the present invention may include CDRs (which are described further herein) from a human or non-human source. The framework of the immunoglobulin can be human, humanized, or non-human, e.g., a murine framework modified to decrease antigenicity in humans, or a synthetic framework, e.g., a consensus sequence.

[0075] The term "antigen-binding portion" of an antibody (or simply "antibody portion," or "portion"), as used herein, refers to a portion of an antibody that specifically binds to an AAH (e.g., HAAH), e.g., a molecule in which one or more immunoglobulin chains is not full length, but which specifically binds to an AAH. Examples of binding portions encompassed within the term "fragment (or antigen-binding portion) or other variant thereof" of an antibody include (i) an Fab fragment, a monovalent fragment consisting of the VLC, VHC, CL and CH1 domains; (ii) a F(ab').sub.2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VHC and CH1 domains; (iv) a Fv fragment consisting of the VLC and VHC domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., Nature 341:544-546, 1989), which consists of a VHC domain; and (vi) an isolated complementarity determining region (CDR) having sufficient framework to specifically bind, e.g., an antigen binding portion of a variable region. An antigen-binding portion of a light chain variable region and an antigen binding portion of a heavy chain variable region, e.g., the two domains of the Fv fragment, VLC and VHC, can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VLC and VHC regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. Science 242:423-426, 1988; and Huston et al. Proc. Natl. Acad. Sci. USA 85:5879-5883, 1988). Such single chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. These antibody portions are obtained using conventional techniques known to those with skill in the art, and the portions are screened for utility in the same manner as are intact antibodies. An Fab fragment can result from cleavage of a tetrameric antibody with papain; Fab' and F(ab')2 fragments can be generated by cleavage with pepsin.

[0076] As used herein, the term "human antibody" includes any antibody in which the framework residues correspond to human germline sequences and the CDRs result from V(D)J recombination and somatic mutations. However, human antibodies may also comprise amino acid residues not encoded in human germline immunoglobulin nucleic acid sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro). It has been demonstrated that in vivo somatic mutation of human variable genes results in mutation of framework residues (see Nat. Immunol. 2:537, 2001). Such an antibody would be termed "human" given its source, despite the framework mutations. Mouse antibody variable domains also contain somatic mutations in framework residues (See Sem. Immunol 8:159, 1996). Consequently, transgenic mice containing the human Ig locus produce antibodies that are commonly referred to as "fully human," even though they possess an average of 4.5 framework mutations (a range of 1-8 in this work: Nat Genet. February 1997;15(2):146-56). Accepted usage therefore indicates that an antibody variable domain gene based on germline sequence but possessing framework mutations introduced by, for example, an in vivo somatic mutational process is termed "human." Thus, the invention encompasses human antibodies that specifically bind AAH (e.g., HAAH, even where those antibodies include mutations (e.g., mutations within the FR) and fragments or other variants thereof (e.g., single chain antibodies that include the VLC and VHC of a multimeric human antibody). For example, the human antibodies of the invention can have 1-8 framework mutation (e.g., about 2, 4, 6, or 8 substitutions, additions, or deletions). Preferably, the sequence of the original human antibody is a human germline sequence.

[0077] Human single-chain antibodies specific for HAAH were produced as described herein. In a particular embodiment, the invention provides antibodies that have specificity for HAAH, and bind to an epitope bound by an antibody described herein (e.g., the antibody encoded by clone 11 or an affinity-matured derivative of clone 11). Antibodies that bind an epitope that overlaps with an epitope bound be an antibody described herein can be identified by their ability to compete with an EGF-like domain for binding to HAAH (e.g., to cells bearing HAAH, such as HAAH transfectants, or H460 tumor cells). The binding site of an anti-AAH antibody can be within the catalytic domain of HAAH.

[0078] The anti-AAH antibodies can be polyclonal or monoclonal. The antibodies and antigen binding portions thereof described herein are useful in therapeutic compositions and regimens, diagnostic compositions and regimens, and in assays requiring an agent that can identify or inhibit an AAH protein. The present invention encompasses an antibody or antigen binding portion thereof for use in therapy (including prophylaxis) or diagnosis (e.g., of particular diseases or conditions such as cancers), and use of such antibodies or antigen binding portions thereof for the manufacture of a medicament for use in treatment of diseases or conditions as described herein.

[0079] Single chain antibodies, and chimeric, humanized or CDR-grafted antibodies, as well as chimeric or CDR-grafted single chain antibodies, comprising portions derived from different species, are also encompassed by the present invention and the term "antibody." The various portions of these antibodies can be joined together chemically by conventional techniques, or can be prepared as contiguous polypeptides using genetic engineering techniques. For example, nucleic acids encoding a chimeric or humanized chain can be expressed to produce a contiguous polypeptide. See, e.g., Cabilly et al, U.S. Pat. No. 4,816,567; Cabilly et al, European Patent No. 0,125,023 B1; Boss et al., U.S. Pat. No.4,816,397; Boss et al., European Patent No. 0,120,694 B1; Neuberger, M. S. et al, WO 86/01533; Neuberger, M. S. et al, European Patent No. 0,194,276 B1; Winter, U.S. Pat. No. 5,225,539; and Winter, European Patent No. 0,239,400 B1. See also, Newman et al, BioTechnology 10: 1455-1460,1992, regarding CDR-graft antibody, and Ladner et al, U.S. Pat. No. 4,946,778 and Bird, R. E. et al., Science 242: 423-426,1988 regarding single chain antibodies.

[0080] In addition, antigen binding portions of antibodies, including fragments of chimeric, humanized, CDR-grafted or single chain antibodies, can also be produced and are within the scope of the present invention. Antigen binding portions of the antibodies retain at least one binding function of the full-length antibody from which they are derived. Preferred antigen binding portions retain an antigen binding function of a corresponding full-length antibody (e.g., specificity for an AAH). Functional fragments can retain the ability of the full-length antibody to inhibit one or more functions characteristic of a AAH, such as AAH's hydroxylase activity. For example, a functional fragment can inhibit hydroxylation of an EGF-like domain. These EGF-like domains contain conserved motifs that form repetitive sequences in diverse proteins, such as clotting factors, extracellular matrix proteins, low-density lipoprotein receptor, Notch homologues or Notch ligand homologues. Any AAH substrate, including those just described, can be used in assays to assess an anti-AAH antibody. Exemplary AAH assay substrates include EGF-IX.sub.1H (Gronke et al., Proc Natl Acad Sci USA 86(10):3609-13, 1989), EGF-X.sub.1H (Gronke et al., J. Biol. Chem. 265:8558-8565,1990), and EGF-Asn (Gronke et al, J. Biol. Chem. 265:8558-8565,1990; Wang et al., J. Biol. Chem. 266:14004-14010,1991).

[0081] For example, antibody portions capable of binding to a AAH or a fragment thereof include Fv, Fab, Fab' and F(ab').sub.2 fragments. Such portions can be produced by enzymatic cleavage or by recombinant techniques. For instance, papain or pepsin cleavage can generate Fab or F(ab').sub.2 fragments, respectively. Antibodies can also be produced in a variety of truncated forms using antibody genes in which one or more stop codons has been introduced upstream of the natural stop site. For example, a chimeric gene encoding a F(ab').sub.2 heavy chain portion can be designed to include DNA sequences encoding the CH.sub.1 domain and hinge region of the heavy chain.

[0082] The invention provides chimeric antibodies that can be prepared as a contiguous polypeptide using genetic engineering techniques (e.g., DNA encoding the protein portions of the chimeric antibody can be expressed to produce a contiguous polypeptide chain). One example of a chimeric antibody of the present invention is an antibody containing one or more antibody chains comprising a CDR (e.g., one or more CDRs of an antibody described herein) and a framework region derived from a light and/or heavy chain of a second antibody (e.g., of human origin; e.g., CDR-grafted antibodies with or without framework changes). In one embodiment, the chimeric antibody can compete with the murine 15C7 or FB50 monoclonal antibody for binding to HAAH. The antigen binding region of the chimeric antibody can be derived from an antibody clone described herein (e.g., clone 11, or a mutant of clone 11; e.g., as in a chimeric antibody comprising CDR1, CDR2 and CDR3 of the clone 11 light chain and CDR1, CDR2 and CDR3 of the clone 11 heavy chain). Chimeric or CDR-grafted single chain antibodies also include humanized immunoglobulin. See, e.g., Cabilly et al, U.S. Pat. No. 4,816,567; Cabilly et al, European Patent No. 0,125,023 B1; Queen et al, European Patent No. 0,451,216 B1; Boss et al, U.S. Pat. No. 4,816,397; Boss et al, European Patent No. 0,120,694 B1; Neuberger et al, WO 86/01533; Neuberger, M. S. et al., European Patent No. 0,194,276 B1; Winter, U.S. Pat. No. 5,225,539; Winter, European Patent No. 0,239,400 B1; Padlan, E. A. et al, European Patent Application No. 0,519,596 A1. See also, Ladner et al, U.S. Pat. No. 4,946,778; Huston, U.S. Pat. No. 5,476,786; and Bird et al, Science 242: 423-426,1988), regarding single chain antibodies.

[0083] Chimeric antibodies can be produced using synthetic and/or recombinant nucleic acids to prepare genes (e.g., cDNA) encoding the desired chimeric chain. For example, nucleic acid (e.g., DNA) sequences coding for variable regions can be constructed using PCR mutagenesis methods to alter DNA sequences encoding an antibody chain, e.g., using methods employed to generate humanized antibodies (see e.g., Kanunan, et al, Nucl. Acids Res. 17: 5404,1989; Sato, et al, Cancer Research 53:851-856,1993; Daugherty, et al, Nucleic Acids Res. 19(9): 2471-2476,1991; and Lewis and Crowe, Gene 101: 297-302,1991). Using these or other suitable methods, variants can also be readily produced. In one embodiment, cloned variable regions can be mutagenized, and sequences encoding variants with the desired specificity can be selected (e.g., from a phage library; see e.g., Krebber et al, U.S. Pat. No. 5,514,548; Hoogenboom et al, WO 93/06213, published Apr. 1, 1993)).

[0084] Other suitable methods of producing or isolating anti-AAH antibodies include, for example, methods that rely upon immunization of transgenic animals (e.g., mice) capable of producing a full repertoire of human antibodies (see e.g., Jakobovits et al, Proc. Natl. Acad. Sci. USA 90: 2551-2555, 1993; Jakobovits et al, Nature 362: 255-258, 1993; Lonberg et al, U.S. Pat. No. 5,545,806; Surani et al., U.S. Pat. No. 5,545,807).

[0085] Antibodies that specifically bind to AAH can be identified by expressing recombinant antibodies in a library and selecting members of the library that bind AAH. The affinity of the selected antibodies for AAH can be further enhanced by affinity-maturing these antibodies, e.g., using PCR mutagenesis, chain shuffling, or CDR shuffling techniques in conjunction with one or more cycles of screening, as described herein. Other methods can also be used to generate anti-AAH antibodies. For example, anti-AAH antibodies can be produced by immunizing animals. A variety of methods have been described for preparing antigen for immunization and for generating monoclonal antibodies from immunized animals (see e.g., Kohler et al, Nature 256:495-497, 1975; Kohler and Milstein, Eur. J. Immunol. 6:511-519, 1976; Milstein et al., Nature 266:550-552,1977; Koprowski et al., U.S. Pat. No. 4,172,124; Harlow, E. and D. Lane, 1988, Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press: Cold Spring Harbor, N.Y.); Current Protocols In Molecular Biology, Vol. 2 (Supplement 27, Summer '94), Ausubel, F. M. et al., Eds., (John Wiley & Sons: New York, N.Y.), Chapter 11, (1991)). Generally, a hybridoma can be produced by fusing a suitable immortal cell line (e.g., a myeloma cell line) with antibody producing cells. The antibody producing cell, preferably those of the spleen or lymph nodes, are obtained from immunized animals. The fused cells (hybridomas) can be isolated using selective culture conditions and cloned by limiting dilution. Cells that produce antibodies with the desired specificity can be selected by a suitable assay (e.g., ELISA).

[0086] The antibody or an antigen-binding portion thereof can include, for example, a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to one of the following sequences: SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:119, SEQ ID NO:121; SEQ ID NO:123, SEQ ID NO:125, SEQ ID NO:127, SEQ ID NO:129, SEQ ID NO:131, SEQ ID NO:133, SEQ ID NO:135, SEQ ID NO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ ID NO:143, SEQ ID NO:145, SEQ ID NO:147, SEQ ID NO:254, SEQ ID NO:256, SEQ ID NO:258, SEQ ID NO:260; SEQ ID NO:262, SEQ ID NO:264, SEQ ID NO:266, SEQ ID NO:268, and SEQ ID NO:270.

[0087] The antibody or an antigen-binding portion thereof can include, for example, a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to one of the following sequences: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:120, SEQ ID NO:122, SEQ ID NO:124, SEQ ID NO:126, SEQ ID NO:128, SEQ ID NO:130, SEQ ID NO:132, SEQ ID NO:134, SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:255, SEQ ID NO:257, SEQ ID NO:259, SEQ ID NO:261; SEQ ID NO:263, SEQ ID NO:265, SEQ ID NO:267, SEQ ID NO:269, and SEQ ID NO:271.

[0088] The antibody or an antigen-binding portion thereof can include, for example, a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:1 and a variable light chain region at least 80% identical to (e.g., 85%, 90%, 95%, 98% or 100%) SEQ ID NO:2; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:3 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:4; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:5 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:6; a variable heavy chain region at least 80% identical to (e.g., 85%, 90%, 95%, 98% or 100%) SEQ ID NO:7 and a variable light chain region at least 80% identical(e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:8; a variable heavy chain region at least 80% identical (e.g., 85%,90%,95%,98% or 100%) to SEQ ID NO:9 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:10; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:11 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQID NO:12; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:13 and avariable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:14; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:15 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:16; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:17 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ IDNO:18; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:19 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:20; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:21 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:22; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:23 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:24; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:25 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:26; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:27 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:28; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:29 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:30; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:119 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:120; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:121 and a variable light chain region at least 80% identical (e.g. 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:122; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:123 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:124; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:125 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:126; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:127 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:128; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:129 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:130; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:131 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:132; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:133 and avariable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:134; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:135 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:136; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:137 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:138; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:139 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:140; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:141 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:142; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:143 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:144; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:145 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:146; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:147 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:148; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:254 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:255; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:256 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:257; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:258 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:259; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:260 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:261; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:262 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:263; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:264 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:265; a variable heavy chain region at least 80% identical (e.g. 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:266 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:267; a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:268 and a variable light chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:269; or a variable heavy chain region at least 80% identical (e.g., 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:270 and a variable light chain region at least 80% identical (e.g. 85%, 90%, 95%, 98% or 100%) to SEQ ID NO:271.

[0089] In various embodiments, the antibody or antigen-binding portion thereof includes a complementarity determining region (CDR) that is at least 40% identical (e.g., 40%, 60%, 80%, 85%, 90%, 95%, 98% or 100%) to any one of SEQ ID NOs. 46-81 or 281-313.

[0090] In various embodiments, the antibody or fragment or other variant thereof includes a complementarity determining region (CDR) that is at least 40% identical (e.g., 40%, 60%, 80%, 85%, 90%, 95%, 98% or 100%) to any one of SEQ ID NOs. 46-51, 54, 56, 82-117, 164-250, or 314-319. The antibody or fragment or other variant thereof can also be, or can include: a variable heavy chain region at least 80% identical to SEQ ID NO:1 and a variable light chain region at least 80% identical to SEQ ID NO:2; a variable heavy chain region at least 80% identical to SEQ ID NO:3 and a variable light chain region at least 80% identical to SEQ ID NO:4; a variable heavy chain region at least 80% identical to SEQ ID NO:5 and a variable light chain region at least 80% identical to SEQ ID NO:6; a variable heavy chain region at least 80% identical to SEQ ID NO:7 and a variable light chain region at least 80% identical to SEQ ID NO:8; a variable heavy chain region at least 80% identical to SEQ ID NO:9 and a variable light chain region at least 80% identical to SEQ ID NO:10; a variable heavy chain region at least 80% identical to SEQ ID NO:11 and a variable light chain region at least 80% identical to SEQ ID NO:12; a variable heavy chain region at least 80% identical to SEQ ID NO:13 and a variable light chain region at least 80% identical to SEQ ID NO:14; a variable heavy chain region at least 80% identical to SEQ ID NO:15 and a variable light chain region at least 80% identical to SEQ ID NO:16; a variable heavy chain region at least 80% identical to SEQ ID NO:17 and a variable light chain region at least 80% identical to SEQ ID NO:18; a variable heavy chain region at least 80% identical to SEQ ID NO:19 and a variable light chain region at least 80% identical to SEQ ID NO:20; a variable heavy chain region at least 80% identical to SEQ ID NO:21 and a variable light chain region at least 80% identical to SEQ ID NO:22; a variable heavy chain region at least 80% identical to SEQ ID NO:23 and a variable light chain region at least 80% identical to SEQ ID NO:24; a variable heavy chain region at least 80% identical to SEQ ID NO:25 and a variable light chain region at least 80% identical to SEQ ID NO:26; a variable heavy chain region at least 80% identical to SEQ ID NO:27 and a variable light chain region at least 80% identical to SEQ ID NO:28; a variable heavy chain region at least 80% identical to SEQ ID NO:29 and a variable light chain region at least 80% identical to SEQ ID NO:30; a variable heavy chain region at least 80% identical to SEQ ID NO:119 and a variable light chain region at least 80% identical to SEQ ID NO:120; avariable heavy chain region at least 80% identical to SEQ ID NO:121 and a variable light chain region at least 80% identical to SEQ ID NO:122; a variable heavy chain region at least 80% identical to SEQ ID NO:123 and a variable light chain region at least 80% identical to SEQ ID NO:124; a variable heavy chain region at least 80% identical to SEQ ID NO:125 and a variable light chain region at least 80% identical to SEQ ID NO:126; a variable heavy chain region at least 80% identical to SEQ ID NO:127 and a variable light chain region at least 80% identical to SEQ ID NO:128; a variable heavy chain region at least 80% identical to SEQ ID NO:129 and a variable light chain region at least 80% identical to SEQ ID NO:130; a variable heavy chain region at least 80% identical to SEQ ID NO:131 and a variable light chain region at least 80% identical to SEQ ID NO:132; a variable heavy chain region at least 80% identical to SEQ ID NO:133 and a variable light chain region at least 80% identical to SEQ ID NO:134; a variableheavy chain region at least 80% identical to SEQ ID NO:135 and avariable light chain region at least 80% identical to SEQ ID NO:136; a variable heavy chain region at least 80% identical to SEQ ID NO:137 and a variable light chain region at least 80% identical to SEQ ID NO:138; a variable heavy chain region at least 80% identical to SEQ ID NO:139 and a variable light chain region at least 80% identical to SEQ ID NO:140; a variable heavy chain region at least 80% identical to SEQ ID NO:141 and a variable light chain region at least 80% identical to SEQ ID NO:142; a variable heavy chain region at least 80% identical to SEQ ID NO:143 and a variable light chain region at least 80% identical to SEQ ID NO:144; a variable heavy chain region at least 80% identical to SEQ ID NO:145 and a variable light chain region at least 80% identical to SEQ ID NO:146; a variable heavy chain region at least 80% identical to SEQ ID NO:147 and a variable light chain region at least 80% identical to SEQ ID NO:148; a variable heavy chain region at least 80% identical to SEQ ID NO:254 and a variable light chain region at least 80% identical to SEQ ID NO:255; a variable heavy chain region at least 80% identical to SEQ ID NO:256 and a variable light chain region at least 80% identical to SEQ ID NO:257; a variable heavy chain region at least 80% identical to SEQ ID NO:258 and a variable light chain region at least 80% identical to SEQ ID NO:259; a variable heavy chain region at least 80% identical to SEQ ID NO:260 and a variable light chain region at least 80% identical to SEQ ID NO:261; a variable heavy chain region at least 80% identical to SEQ ID NO:262 and a variable light chain region at least 80% identical to SEQ ID NO:263; a variable heavy chain region at least 80% identical to SEQ ID NO:264 and a variable light chain region at least 80% identical to SEQ ID NO:265; a variable heavy chain region at least 80% identical to SEQ ID NO:266 and a variable light chain region at least 80% identical to SEQ ID NO:267; a variable heavy chain region at least 80% identical to SEQ ID NO:268 and a variable light chain region at least 80% identical to SEQ ID NO:269; or a variable heavy chain region at least 80% identical to SEQ ID NO:270 and a variable light chain region at least 80% identical to SEQ ID NO:271.

[0091] As used herein, the term "substantially identical" (or "substantially homologous") refers to a first amino acid or nucleotide sequence that contains a sufficient number of identical or equivalent (e.g., with a similar side chain, e.g., conserved amino acid substitutions) amino acid residues or nucleotides to a second amino acid or nucleotide sequence such that the first and second amino acid or nucleotide sequences have similar activities. In the case of antibodies, the second antibody has the same specificity and has at least 50% of the affinity of the first antibody.

[0092] Calculations of "homology" or "identity" between two sequences can be performed as follows. The sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In different embodiments, the length of a reference sequence aligned for comparison purposes is at least 50% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

[0093] The comparison of sequences and determination of percent homology between two sequences can be accomplished using a mathematical algorithm. The percent homology between two amino acid sequences is determined using the Needleman and Wunsch, J. Mol. Biol. 48:444-453, 1970, algorithm which has been incorporated into the GAP program in the GCG software package, using a BLOSUM 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

[0094] As used herein, the term "hybridizes under low stringency, medium stringency, high stringency, or very high stringency conditions" describes conditions for hybridization and washing. Guidance for performing hybridization reactions can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. 6.3.1-6.3.6, 1989, which is incorporated herein by reference. Aqueous and nonaqueous methods are described in that reference and either can be used. Specific hybridization conditions referred to herein are as follows: 1) low stringency hybridization conditions in 6.times. sodium chloride/sodium citrate (SSC) at about 45.degree. C., followed by two washes in 0.2.times.SSC, 0.1% SDS at least at 50.degree. C. (the temperature of the washes can be increased to 55.degree. C. for low stringency conditions); 2) medium stringency hybridization conditions in 6.times.SSC at about 45.degree. C., followed by one or more washes in 0.2.times.SSC, 0.1% SDS at 60.degree. C.; 3) high stringency hybridization conditions in 6.times.SSC at about 45.degree. C., followed by one or more washes in 0.2.times.SSC, 0.1% SDS at 65.degree. C.; and 4) very high stringency hybridization conditions are 0.5 M sodium phosphate, 7% SDS at 65.degree. C., followed by one or more washes at 0.2.times.SSC, 1% SDS at 65.degree. C.

[0095] It is understood that the antibodies and antigen binding portions thereof of the invention may have additional conservative or non-essential amino acid substitutions (a "non-essential" amino acid residue is a residue that can be altered from the wild-type sequence of a polypeptide, such as a binding agent, e.g., an antibody, without substantially altering a biological activity, whereas an "essential" amino acid residue results in such a change).

[0096] Whether or not a particular substitution will be tolerated, i.e., will not adversely affect desired biological properties, such as binding activity, can be determined as described in Bowie et al., Science, 247:1306-1310, 1990. A "conservative amino acid substitution" is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g. tyrosine, phenylalanine, tryptophan, histidine).

[0097] As described herein, antibodies of the present invention can inhibit hydroxylase activity of AAH and/or inhibit a function associated with hydroxylase activity, such as cell motility. As discussed below, various methods can be used to assess inhibition of AAH activity and/or function associated with the activity.

Binding Assays

[0098] As used herein "mammalian AAH protein" refers to naturally occurring or endogenous mammalian AAH proteins and to proteins having an amino acid sequence which is the same as that of a naturally occurring or endogenous corresponding mammalian AAH protein (e.g., recombinant proteins). Accordingly, as defined herein, the term includes polymorphic or allelic variants, and other isoforms of a mammalian AAH (e.g. produced by alternative splicing or other cellular processes), and modified or unmodified forms of the foregoing (e.g. glycosylated, unglycosylated). AAH proteins can be isolated and/or recombinant proteins (including synthetically produced proteins). Naturally occurring or endogenous mammalian AAH can be recovered or isolated from a source which naturally produces AAH, for example, a tumor cell.

[0099] "Functional variants" of AAH proteins include functional fragments, functional mutant proteins, and/or functional fusion proteins (e.g., produced via mutagenesis and/or recombinant techniques). Generally, fragments or portions of AAH proteins include those having a deletion (i.e., one or more deletions) of an amino acid (i.e., one or more amino acids) relative to the mature mammalian AAH (such as N-terminal, C-terminal or internal deletions).

[0100] Generally, mutants of AAH proteins include natural or artificial variants of an AAH protein differing by the addition, deletion and/or substitution of one or more contiguous or non-contiguous amino acid residues. Such mutations can be in a conserved region or nonconserved region, extracellular, cytoplasmic, or transmembrane region, for example.

[0101] A "functional fragment or portion", "functional mutant" and/or "functional fusion protein" of an AAH protein refers to an isolated and/or recombinant protein or polypeptide which has at least one function characteristic of a AAH protein as described herein, such as a hydroxylase activity.

[0102] A composition comprising an isolated and/or recombinant mammalian AAH or portion thereof can be maintained under conditions suitable for binding, the receptor is contacted with an antibody to be tested, and binding is detected or measured. In one embodiment, a receptor protein can be expressed in cells which express AAH or in cells stably or transiently transfected with a construct comprising a nucleic acid sequence which encodes an AAH or portion thereof. The cells are maintained under conditions appropriate for expression of receptor. The cells are contacted with an antibody under conditions suitable for binding (e.g., in a suitable binding buffer), and binding is detected by standard techniques. To measure binding, the extent of binding can be determined relative to a suitable control (e.g., compared with background determined in the absence of antibody, compared with binding of a second antibody (i.e., a standard), compared with binding of antibody to untransfected cells). A cellular fraction, such as a membrane fraction, containing AAH can be used in lieu of whole cells.

[0103] In one embodiment, the antibody is labeled with a suitable label (e.g. fluorescent label, isotope label, enzyme label), and binding is determined by detection of the label. In another embodiment, bound antibody can be detected by labeled second antibody. Specificity of binding can be assessed by competition or displacement, for example, using unlabeled antibody or a ligand as competitor.

[0104] Binding inhibition assays can also be used to identify antibodies that bind AAH and inhibit binding of another compound such as an EGF-like domain. For example, a binding assay can be conducted in which a reduction in the binding of an EGF-like domain (in the absence of an antibody), as compared to binding of the ligand in the presence of the antibody, is detected or measured. AAH can be contacted with a protein containing an EGF-like domain and antibody simultaneously, or one after the other, in either order. A reduction in the extent of binding of the protein in the presence of the antibody, is indicative of inhibition of binding by the antibody. For example, binding of the EGF-like domain could be decreased or abolished.

[0105] Other methods of identifying the presence of an antibody which binds AAH are available, such as other suitable binding assays, or methods which monitor events which are triggered by AAH activity, e.g., cellular transformation, or cell motility.

[0106] It will be understood that the inhibitory effect of antibodies of the present invention can be assessed in a binding inhibition assay. Competition between antibodies for binding can also be assessed in the method. Antibodies which are identified in this manner can be further assessed to determine whether, subsequent to binding, they act to inhibit other functions of AAH and/or to assess their therapeutic utility.

Assays to Determine Antibody Activity

[0107] AAH hydroxylates the .beta. carbon of aspartic acid or asparagines residues in epidermal growth factor (EGF)-like domains of proteins in the presence of ferrous iron. EGF-like domains, which contain a conserved CX.sub.7CX.sub.4CX.sub.10CXCX.sub.8C sequence, are present in many diverse proteins, such as clotting factors, extracellular matrix proteins, low density lipoprotein receptor, Notch homologs, and Notch ligand homologs. Hydroxylation of AAH substrates can involve direct detection of hydroxylase activity, or can be measured by indirectly, e.g., using assays that detect a biological activity downstream of hydroxylation.

[0108] An assay to determine whether a human anti-AAH antibody inhibits AAH activity can be performed by comparing the level of hydroxylation in an enzymatic reaction in which the test antibody is present compared to a parallel reaction in the absence of a compound, or compared to a predetermined control value. Standard hydroxylase assays are known. See, e.g., Lavaissiere et al, J. Clin. Invest. 98:1313-1323,1996; Jia et al., J. Biol. Chem. 267:14322-14327,1992; Wang et al., J. Biol. Chem. 266:14004-14010,1991; or Gronke et al., J. Biol. Chem. 265:8558-8565,1990. Hydroxylase activity can be measured using carbon dioxide (.sup.14CO.sub.2 capture assay) in a microtiter plate (Zhang et al, Anal Biochem. 271:137-142, 1999).

[0109] Modulation of AAH activity can be determined by examining biochemical effects downstream of hydroxylation, including effects on specific substrates, and effects on cellular processes. AAH activity increases cell motility, proliferation, survival, and cell cycle progression. Inhibition of AAH activity by an antibody can be determined by detecting a reduction in one of these processes in the presence of the antibody.

[0110] Modulation of cell motility can be assayed, e.g., using a motility assay. Generally, motility assays monitor the directional movement or migration of a suitable cell (such as a tumor cell) into or through a barrier (e.g., a filter), toward increased levels of a compound (e.g., a growth factor or other polypeptide), from a first surface of the barrier toward an opposite second surface. Membranes or filters provide convenient barriers, such that the directional movement or migration of a suitable cell into or through a filter, toward increased levels of a compound, from a first surface of the filter toward an opposite second surface of the filter, is monitored. One can detect or measure inhibition of the migration of cells in a suitable container from a first chamber into or through a microporous membrane into a second chamber which contains an antibody to be tested, and which is divided from the first chamber by the membrane. A suitable membrane, having a suitable pore size for monitoring specific migration in response to compound, including, for example, nitrocellulose, polycarbonate, is selected. For example, pore sizes of about 3-8 microns can be used.

[0111] To assess migration and inhibition of migration, the number of cells crossing the filter that remain adherent to the second surface of the filter, and/or the number of cells that accumulate in the second chamber can be determined using standard techniques (e.g., microscopy). In one embodiment, the cells are labeled with a detectable label (e.g., radioisotope, fluorescent label, antigen or epitope label), and migration can be assessed in the presence and absence of the antibody by determining the presence of the label adherent to the membrane and/or present in the second chamber using an appropriate method (e.g., by detecting radioactivity, fluorescence, immunoassay). The extent of migration induced by an antibody agonist can be determined relative to a suitable control (e.g., compared to background migration determined in the absence of the antibody, compared to the extent of migration induced by a second compound (i.e., a standard), compared with migration of untransfected cells induced by the antibody). A reduction in the extent of migration induced by the compound (e.g., calf serum) in the presence of the antibody is indicative of inhibitory activity.

[0112] In one embodiment, cells are placed in the upper chamber of a Boyden chamber-type culture insert in serum-free medium. Medium supplemented with 1%-2% fetal calf serum is placed in the lower chamber to provide a stimulus for migration. Cells are incubated for approximately 4 hours to allow migration to occur. Cell numbers in the upper and lower chambers are quantified. Viable cells in each chamber can be quantified using an ATP monitoring system such as ATPLite.TM. (Perkin Elmer.RTM.; see, e.g., Sepe, et al. Lab. Invest. 82:881-891, 2002 for a description of a motility assay).

[0113] AAH overexpression is linked to cellular proliferation and malignant transformation. Inhibition of AAH activity can be assayed by measuring cellular characteristics of malignant phenotypes, such as transformation, anchorage-independent cell growth, and tumorigenicity in nude mice. Transformation can be assessed by transfecting NIH 3T3 cells with AAH and observing the number of transformed foci (Copeland and Cooper, Cell 16(2):347-56, 1979). Anchorage-independent cell growth can be assayed by transfecting cells with AAH, isolating transfectants, and suspending transfectants in complete medium containing 0.4% low-melting agarose laid over a bottom layer of medium containing 0.53% low-melting agarose.

[0114] To assay tumorigenicity in vivo, AAH-transfected clones (or AAH-expressing tumor cells) are injected into nude mice. In a typical assay, approximately 1-10 million cells are injected subcutaneously. After growth for 1 week-1 month, animals are sacrificed, and tumors are removed and weighed. Antibodies can be tested by injection into animals implanted with AAH-transfectants or tumor cells and comparing growth of the implanted cells to growth of cells in animals receiving a control injection (e.g., saline, or a non-specific antibody).

[0115] Antibodies with inhibitory activity may reduce efficiency of cell growth, transformation, and tumorigenicity in these assays.

Diagnostic and Therapeutic Applications

[0116] The antibodies of the present invention are useful in a variety of applications, including research, diagnostic and therapeutic applications. In one embodiment, the antibodies are labeled with a suitable label (e.g., fluorescent label, chemiluminescent label, isotope label, epitope or enzyme label).

[0117] Overexpression of HAAH is associated with malignant transformation. Antibodies or antigen binding portions thereof that block and/or inhibit the activity of HAAH can be used to inhibit cell transformation and/or to diagnose transformed cells. Accordingly, the present invention provides a method of inhibiting AAH activity of a cell which expresses an AAH or portion thereof, comprising contacting the cell with an effective amount of an antibody or antigen binding portion thereof which binds to the AAH or a portion of the AAH. The cell can be a cell of a subject (e.g., a tumor cell), and the antibody or antigen binding portion thereof can be administered to the subject in vivo. Therapeutic use of an antibody or antigen binding portion thereof includes prophylactic use (e.g., for treatment of a patient who may be at risk for developing a cancer).

[0118] The anti-AAH antibody or antigen binding portion thereof can be administered in combination with one or more other therapeutic agents such as an anti-cancer agent. Nonlimiting examples of anti-cancer agents include, e.g., antimicrotubule agents, topoisomerase inhibitors, antimetabolites, mitotic inhibitors, alkylating agents, intercalating agents, agents capable of interfering with a signal transduction pathway, agents that promote apoptosis (including cell death genes), radioactive compounds, and antibodies against other tumor-associated antigens (including naked antibodies, immunotoxins and radioconjugates). Examples of the particular classes of anti-cancer agents are provided in detail as follows: antitubulin/antimicrotubule, e.g., paclitaxel, vincristine, vinblastine, vindesine, vinorelbin, taxotere; topoisomerase I inhibitors, e.g., topotecan, camptothecin, doxorubicin, etoposide, mitoxantrone, daunorubicin, idarubicin, teniposide, amsacrine, epirubicin, merbarone, piroxantrone hydrochloride; antimetabolites, e.g., 5-fluorouracil (5-FU), methotrexate, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, cytarabine/Ara-C, trimetrexate, gemcitabine, acivicin, alanosine, pyrazofurin, N-Phosphoracetyl-L-Asparate (PALA), pentostatin, 5-azacitidine, 5-Aza 2'-deoxycytidine, ara-A, cladribine, 5-fluorouridine, FUDR, tiazofurin, N-[5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-- 2-thenoyl]-L-glutamic acid; alkylating agents, e.g., cisplatin, carboplatin, mitomycin C, melphalan, thiotepa, busulfan, chlorambucil, plicamycin, dacarbazine, ifosfamide phosphate, cyclophosphamide, nitrogen mustard, uracil mustard, pipobroman, 4-ipomeanol; agents acting via other mechanisms of action, e.g., dihydrolenperone, spiromustine, and desipeptide; biological response modifiers, e.g., to enhance anti-tumor responses, such as interferon; apoptotic agents, such as actinomycin D; and anti-hormones, for example anti-estrogens such as tamoxifen or, for example antiandrogens such as 4'-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3'-(trifluorometh- yl)propionanilide.

[0119] The anti-AAH antibodies of the present invention also have value in diagnostic applications. An anti-AAH antibody can be used to monitor growth and/or metastasis of a tumor in vivo, and may be used as a diagnostic indicator of disease stage. Human antibodies are not immunogenic in humans, and therefore human antibodies may be more appropriate for in vivo diagnostic applications than mouse antibodies.

[0120] For diagnostic purposes, the antibodies or antigen binding portions can be labeled or unlabeled. Typically, diagnostic assays entail detecting the formation of a complex resulting from the binding of an antibody or portion to AAH. The antibodies or portions can be directly labeled. A variety of labels can be employed, including, but not limited to, radionuclides, fluorescers, enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors and ligands (e.g., biotin, haptens, and the like). Numerous appropriate immunoassays are known to the skilled artisan (see, for example, U.S. Pat. Nos. 3,817,827; 3,850,752; 3,901,654 and 4,098,876). Unlabeled antibodies or fragments can also be used in combination with another (i.e., one or more) suitable reagent that can be used to detect antibody, such as a labeled antibody (e.g., a second antibody) reactive with the first antibody (e.g., anti-idiotype antibodies or other antibodies that are specific for the unlabeled immunoglobulin) or other suitable reagent (e.g., labeled protein A).

[0121] Kits for use in detecting the presence of AAH in a biological sample can also be prepared. Such kits will include an antibody or antigen binding portion thereof which binds to an AAH protein or portion of said receptor, as well as one or more ancillary reagents suitable for detecting the presence of a complex between the antibody or portion and AAH or portion thereof. The antibody compositions of the present invention can be provided in lyophilized form, either alone or in combination with additional antibodies specific for other epitopes. The antibodies, whether labeled or unlabeled, can be included in the kits with adjunct ingredients (e.g., buffers, such as Tris, phosphate and carbonate, stabilizers, excipients, biocides and/or inert proteins, e.g., bovine serum albumin). For example, the antibodies can be provided as a lyophilized mixture with the adjunct ingredients, or the adjunct ingredients can be separately provided for combination by the user. Where a second antibody capable of binding to the monoclonal antibody is employed, such antibody can be provided in the kit, for instance in a separate vial or container. The second antibody, if present, is typically labeled, and can be formulated in an analogous manner with the antibody formulations described above.

[0122] Similarly, the present invention also relates to a method of detecting and/or quantifying expression of AAH or portion of the enzyme by a cell, in which a composition comprising a cell or fraction thereof (e.g., membrane fraction) is contacted with an antibody or functional portion thereof which binds to AAH or portion of AAH (e.g., the catalytic domain) under conditions appropriate for binding of the antibody, and antibody binding is monitored. Detection of the antibody, indicative of the formation of a complex between antibody and AAH or a portion thereof, indicates the presence of AAH. Binding of antibody to the cell can be determined as described above under the heading "Binding Assays", for example. The method can be used to detect expression of AAH in cells from an individual (e.g., in a tumor biopsy sample). A quantitative expression of AAH on the surface of tumor cells can be evaluated, for instance, by flow cytometry, and the staining intensity can be correlated with disease progression or risk.

[0123] AAH has a role in cell motility, and so anti-AAH antibodies can be used to inhibit (reduce or prevent) tumor growth or metastasis. Accordingly, the antibodies of the present invention can also be used to modulate AAH function in research and therapeutic applications. For instance, the antibodies described herein can act as inhibitors to inhibit (reduce or prevent) (a) binding (e.g., of an EGF-like domain of a protein) to AAH, (b) a receptor signaling function mediated by AAH, and/or (c) a stimulatory function (e.g., of a substrate of AAH or an AAH-pathway). Antibodies which act as inhibitors of receptor function can block AAH binding directly or indirectly (e.g., by causing a conformational change). Thus, the present invention provides a method of inhibiting cell motility in a mammal (e.g., a human patient), comprising administering to the mammal an effective amount of an antibody or antigen binding portion thereof which binds to a AAH or portion of AAH. Diseases which can be treated according to the method include cancers and can result in amelioration of the disease state. The cancers which can be treated include, but are not limited to solid tumors, soft tissue tumors, and metastatic lesions. Examples of solid tumors include malignancies, e.g., sarcomas, adenocarcinomas, and carcinomas, of the various organ systems, such as those affecting lung, breast, lymphoid, gastrointestinal (e.g., colon), and genitourinary tract (e.g., renal, urothelial cells), liver, pharynx, prostate, ovary, cholangiocarcinomas, as well as adenocarcinomas which include malignancies such as most colon cancers, rectal cancer, renal-cell carcinoma, liver cancer, non-small cell carcinoma of the lung, cancer of the small intestine, neuroectodermal tumors, and so forth. Metastatic lesions of the aforementioned cancers can also be treated or prevented using the methods and compositions of the invention.

[0124] The subject method can be useful in treating malignancies of the various organ systems, such as those affecting lung, breast, lymphoid, gastrointestinal (e.g., colon), and genitourinary tract, prostate, ovary, pharynx, as well as adenocarcinomas which include malignancies such as most colon cancers, renal-cell carcinoma, prostate cancer and/or testicular tumors, non-small cell carcinoma of the lung, cancer of the small intestine and cancer of the esophagus. Exemplary solid tumors that can be treated include: fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lyrnphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, non-small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, and retinoblastoma.

[0125] Patients amenable to treatment with the anti-AAH antibodies described herein may be described as having cancer or a "carcinoma." Carcinomas are recognized by those of ordinary skill in the art as malignancies of epithelial or endocrine tissues including respiratory system carcinomas, gastrointestinal system carcinomas, genitourinary system carcinomas, testicular carcinomas, breast carcinomas, prostatic carcinomas, endocrine system carcinomas, and melanomas. Exemplary carcinomas include those forming from tissue of the cervix, lung, prostate, breast, head and neck, colon and ovary. The term also includes carcinosarcomas, e.g., which include malignant tumors composed of carcinomatous and sarcomatous tissues. An "adenocarcinoma" refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures. The term "sarcoma" is recognized by those skilled in the art and refers to malignant tumors of mesenchymal derivation.

[0126] The methods of the invention can also be used to inhibit the proliferation of hyperplastic/neoplastic cells of hematopoietic origin, e.g., arising from myeloid, lymphoid or erythroid lineages, or precursor cells thereof. For instance, the present invention contemplates the treatment of various mycloid disorders including, but not limited to, acute promyeloid leukemia (APML), acute myclogenous leukemia (AML) and chronic myelogenous leukemia (CML) (reviewed in Vaickus, Crit Rev. in Oncol/Hemotol. 11:267-97, 1991). Lymphoid malignancies which may be treated by the subject method include, but are not limited to acute lymphoblastic leukemia (ALL), which includes B-lineage ALL and T-lineage ALL, chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), hairy cell leukemia (HLL) and Waldenstrom's macroglobulinemia (WM). Additional forms of malignant lymphomas contemplated by the treatment method of the present invention include, but are not limited to, non-Hodgkin's lymphoma and variants thereof, peripheral T-cell lymphomas, adult T-cell leukemia/lymphoma (ATL), cutaneous T-cell lymphoma (CTCL), large granular lymphocytic leukemia (LGF) and Hodgkin's disease.

Modes of Administration

[0127] According to the method, one or more antibodies or antigen binding portions thereof can be administered to the host by an appropriate route, either alone or in combination with (before, simultaneous with, or after) another drug. For example, the antibodies of the present invention can also be used in combination with other monoclonal or polyclonal antibodies or with chemotherapeutic treatments.

[0128] An effective amount of an antibody (i.e., one or more antibodies or fragments) is administered. An effective amount is an amount sufficient to achieve the desired therapeutic effect, under the conditions of administration, such as an amount sufficient for inhibition of an AAH function, and thereby, inhibition of a tumor cell.

[0129] A variety of routes of administration are possible including, but not necessarily limited to, parenteral (e.g., intravenous, intraarterial, intramuscular, subcutaneous injection), oral, dietary, topical, inhalation (e.g., intrabronchial, intranasal or oral inhalation, intranasal drops), depending on the disease or condition to be treated. Other suitable methods of administration can also include rechargeable or biodegradable devices and slow release polymeric devices. The pharmaceutical compositions described herein can also be administered as part of a combinatorial therapy with other agents.

[0130] Formulation of an antibody or portion thereof to be administered will vary according to the route of administration selected (e.g., solution, emulsion, capsule). An appropriate pharmaceutical composition comprising an antibody or antigen binding portion thereof to be administered can be prepared in a physiologically acceptable vehicle or carrier. A mixture of antibodies and/or portions can also be used. For solutions or emulsions, suitable carriers include, for example, aqueous or alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles can include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils. A variety of appropriate aqueous carriers are known to the skilled artisan, including water, buffered water, buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), dextrose solution and glycine. Intravenous vehicles can include various additives, preservatives, or fluid, nutrient or electrolyte replenishers (See, generally, Remington's Pharmaceutical Science, 16th Edition, Mack, Ed. 1980). The compositions can optionally contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents and toxicity adjusting agents, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride and sodium lactate. The antibodies can be lyophilized for storage and reconstituted in a suitable carrier prior to use according to art-known lyophilization and reconstitution techniques. The optimum concentration of the active ingredient(s) in the chosen medium can be determined empirically, according to procedures well known to the skilled artisan, and will depend on the ultimate pharmaceutical formulation desired. For inhalation, the compound can be solubilized and loaded into a suitable dispenser for administration (e.g., an atomizer, nebulizer or pressurized aerosol dispenser).

Examples

[0131] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

Example 1

Library Screen for Anti-HAAH Antibodies

[0132] A yeast surface display technique (Boder and Wittrup, Nat Biotechnol. 15(6):553-7, 1997) was used to screen a naive human single-chain Fv library for anti-HAAH Fv fragments. This library was prepared as described in Feldhaus et al. (Nature Biotech. 21:163-170, 2003). Briefly, human antibody variable genes were cloned by PCR from commercially available spleen and lymph node poly(A) mRNA pooled from 58 adults. Primers to IgG, IgM, K and X were used for first-strand cDNA synthesis. Separate VH and VL libraries were constructed, then assembled together in single-chain Fv (scFv) format by overlap extension PCR. The scFv library was then suboloned for expression as an Aga2p fusion on the yeast surface. The Aga2p-scFv fusion protein is depicted schematically in FIG. 1. A library of approximately 10.sup.9 scFv fragments was expressed in yeast.

[0133] The library was screened for binding to a recombinant form of HAAH comprising the extracellular domain as described in Boder and Wittrup (Biotechnol Prog. 14(1):55-62, 1998). Briefly, yeast cells expressing Aga2p-scFv fusions are assayed for binding to fluorescent HAAH by flow cytometry (FACS). Binders are selected by sorting and re-analyzed. Six rounds of screening for binding in 300 nM HAAH allowed the identification of 16 unique clones. FACS analysis of binding of three unique clones to HAAH is depicted in FIG. 2. FACS analysis of binding of 8 unique clones to the catalytic domain of HAAH is depicted in FIG. 3. Of these clones, eight bound to the catalytic domain of HAAH.

Example 2

Binding Activity of Anti-HAAH Antibodies

[0134] The affinity of various scFv clones for HAAH was determined by performing a titration in which cells displaying a given scFv are labeled with varying concentrations of unlabeled HAAH, then varying concentrations of labeled HAAH, and detecting the labeling intensity by FACS. The affinities for two representative anti-HAAH scFv clones are depicted in FIG. 4. Clone 5-2 displayed an affinity for HAAH of approximately 500 nm. Clone 6-12 displayed an affinity for HAAH of approximately 150 nM.

[0135] One antibody clone, clone 27, was isolated which bound to full-length HAAH, but not to the catalytic domain of HAAH. FACS analysis of binding of this scFv clone to the catalytic domain of HAAH and full-length HAAH is depicted in FIG. 5.

Example 3

Binding by Soluble Anti-HAAH scFv Fragments

[0136] The clones that bound to the catalytic domain were expressed in soluble form and analyzed for binding and other biological activities. Soluble scFv fragments of six unique clones were analyzed for binding to the catalytic domain of HAAH (FIG. 6), and full-length HAAH (FIG. 7). ScFv fragments were present at 1-2 .mu.M. Clone 11 showed the highest level of non-specific binding in these assays.

[0137] Next, binding by 3 unique scFv clones to H460 human lung carcinoma cells, which express HAAH, was assessed. ScFv fragments were tested at approximately 1 .mu.M. The results of these experiments are depicted in FIG. 8. Clone 11 scFv also showed the highest level of binding in these assays. Clone 12 also bound H460 cells.

Example 4

Inhibition of Cell Motility by Soluble Anti-HAAH scFv Fragments

[0138] Five unique soluble scFv clones and one mouse IgG, 15C7, were tested for inhibition of H460 cell motility (FIG. 9). Cells were placed in serum-free medium in a filter cup with a pore size of approximately 8 microns. The filter cup was placed into medium containing serum, and cells were incubated to allow migration through the filter. After the incubation, cell number on each side of the filter was determined by staining with crystal violet and counting using a microscope and a hemocytometer. Sixty percent of H460 cells were motile in the absence of antibody. Antibodies were present at 2 .mu.M in the motility assays. 15C7 blocked motility by 100%. Only about 10% of cells were motile in the presence of clone 11 scFv. Clones 10 and 13 also showed modest inhibition of cell motility. Inhibition of cell growth by clone 11 was also assayed. Clone 11 did not demonstrate inhibition of H460 growth at 2 .mu.M (data not shown).

Example 5

Error Prone PCR for Affinity Maturation of Anti-HAAH scFv Fragments

[0139] Next, nucleic acids encoding HAAH-binding scFv were mutagenized and reselected for binding in order to produce "affinity-matured" antibodies in vitro. Mutagenesis was performed by subjecting the scFv-encoding DNA to error prone PCR using nucleotide analogs as described in Zaccolo et al. (J. Mol. Biol., 255:589-603, 1996) and Zaccolo et al. (J. Mol. Biol. 285:775-783, 1999). The initial post-mutagenesis (pre-affinity selection) library had approximately 3.times.10.sup.6 members. Selected clones isolated from the library after mutagenesis and which contained full-length open reading frames were sequenced. The mean number of amino acid sequence changes in clones derived from clone 11, relative to the input sequences, was 7.+-.8 (out of 266 amino acids total).

[0140] The mutagenized nucleic acids were re-expressed with yeast surface display and screened for binding using FACS as described above. DNA encoding binders was isolated, re-mutagenized, re-expressed and screened, and so forth for six rounds. Five of the six rounds were performed with sequentially decreasing concentrations of HAAH, starting at 1 .mu.M and ending with 200 mM. One round was performed with detection antibodies in the absence of HAAH to deplete clones that bound to the detection antibody. The mean number of amino acid sequence changes in seven clones derived from clone 11 after this sorting step was 23.+-.16, relative to the input sequences. The number of mutations in germline-encoded framework regions ranged from 8 to 35, as compared to 4 mutations in the original clone (relative to the germline sequence).

[0141] The binding of an improved clone 11 scFv mutant was assayed for binding to the catalytic domain of HAAH (0.5 .mu.M) (FIG. 10). The amino acid sequence of this improved clone is depicted in FIG. 11. Amino acid changes relative to the original clone 11 are shown in bold-faced type and underlined.

[0142] HAAH binding of clone 11 mutants from successive rounds of mutagenesis was assayed by FACS as described above. The level of expression of scFv on yeast cells was analyzed in parallel. The results of these experiments are depicted in FIG. 12. As shown in FIG. 12, the mutants produced after four rounds of screening ("1.sup.st round mutants") displayed a higher level of binding to HAAH as compared to original clone 11. One clone, 11m1-2, also displayed a uniformly higher level of binding as compared to original clone 11. Binding of other affinity matured clone 11 mutants is shown in FIG. 13. The dissociation rate constant of the best-improved clones (e.g., 11m1-2) was determined to be 220.+-.60 nM, an improvement of two orders of magnitude over the initial, non-mutagenized scFv from which it was derived.

[0143] Five out of seven affinity matured clones derived from clone 11 had a mutation of cysteine to arginine or tyrosine at position H22 (according to Kabat numbering) in the heavy chain. This cysteine forms an intra-domain disulfide bond. Of the five clones with this mutation, three of them have an addition of a cysteine, with the occurrence of the cysteine residue in either the heavy chain framework region 3 (FW3) or in the light chain CDR3. Of the two clones that did not have a cysteine H22 mutation, one lacked additional cysteines. The other had a replacement of a tyrosine with a cysteine in the heavy chain CDR3 loop, four residues away from cysteine H22. The frequent cysteine mutations may indicate that relaxation of the heavy chain structure can lead to improved binding.

[0144] Affinity maturation was also performed with clone 13. Binding of a clone 13 mutant, 13m1, to the catalytic domain of HAAH is shown in FIG. 14. 13m1 was generated after five cycles of mutagenesis/screening. The amino acid sequence of this mutant is depicted in FIG. 15. Amino acid changes relative to the original clone 13 are bold and underlined. Binding of successive mutants of clone 13 was assayed as described for the clone 11 mutants. As shown in FIG. 16, clone 13m1, and a clone generated from 13m1, 12m2-3, both displayed an enhanced level of binding relative to original clone 13.

Example 6

Chain Shuffling for Affinity Maturation of Anti-HAAH scFv Fragments

[0145] Chain shuffling is a mutagenesis technique whereby an entire antibody chain or portion of a chain is recombined with other chains. In order to generate anti-HAAH scFv fragments with increased affinity for HAAH, a library was constructed by shuffling heavy chains from the naive library against the wild type anti-HAAH light chain (to preserve the light chain sequence and link it with different heavy chain sequences). Specifically, heavy chain fragments from the human naive library were extracted by restriction digestion with NheI and BamHI and ligated into a NheI-BamHI-digested yeast display plasmid containing the light chain of the wild type scFv as depicted in FIG. 17. The initial library generated by this process had approximately 1.2.times.10.sup.5 members. Five clones from the initial library were sequenced, and all had heavy chains which were different from the initial heavy chain. FIG. 18A contains a chart listing the heavy chains and light chains in each clone.

[0146] Next, six rounds of FACS screening were performed in the presence of decreasing concentrations of antigen (from 800 nM to 500 nM of HAAH) to screen for improved binders. An additional step of screening in the absence HAAH was performed to eliminate clones binding to the detection antibodies. The scFv display level (X-axis) and HAAH binding (Y-axis) of yeast cells from the pool of mutagenized cells was analyzed by FACS. FIG. 18B shows that clones with high levels of high-affinity scFv fragments were present in the pool.

[0147] Twenty clones were sequenced after the sixth round of screening. Eleven of the clones that were sequenced were unique and each contained a different heavy chain from the wild type scFv. The scFv display level (X-axis) and HAAH binding (Y-axis) of these clones is shown in FIGS. 19A-19K. The dissociation constants for two selected clones, LLm11 and LLm13, were determined to be 240.+-.70 nM and 160.+-.50 nM respectively, which is an improvement of over two orders of magnitude with respect to wild type.

[0148] The framework sequences of the clones generated by this technique had minimal deviation from the germline framework sequences (on average 4.+-.2 mutations from heavy chain germline framework), rendering them less likely to be immunogenic than clones with a greater number of framework mutations. Because all these clones have different heavy chain CDR3, they may have slightly different binding specificities. Multiple binders having different specificities against the same target can be readily screened for secondary desired properties such as minimal binding towards particular human tissues.

Example 7

Affinity Maturation of Anti-HAAH scFv Fragments Using CDR Shuffling

[0149] A third technique, CDR shuffling (also referred to as domain shuffling) was used to generate scFv clones with enhanced affinity for HAAH. To perform CDR shuffling, yeast cells are co-transformed with an acceptor vector and multiple linear insert sequences. The acceptor vector and the insert sequences are designed to have homologous sequences at one or both of their 5' and 3' ends. Homologous recombination between these sites, and at random sites within the gene of interest (e.g., antibody chain) generates "shuffled" products. Further details for performing CDR shuffling are described in Swers et al. (Nucl. Acids Res. 32(3):e36, 2004).

[0150] CDR shuffling was performed with DNA encoding the wild type clone 11 scFv fragment as the acceptor vector, and sequences from a human scFv library as the insert sequences (FIG. 21). The insert sequences had a light chain identical to the clone 11 light chain and the insert sequences of each of three libraries contained regions corresponding to HC FW1 to CDR1, HC FW1 to CDR2, or HC FW1 to CDR3, replaced with naive heavy chain gene sequences from the same VH family as the original clone. The libraries containing replacements of HC FW1 to CDR1 and HC FW1 to CDR2 regions result in a mutagenesis event similar to receptor editing, which naturally occurs in B cells in vivo.

[0151] The libraries with HC FW1 to CDR1, HC FW1 to CDR2, or HC FW1 to CDR3 inserts contained 1.1.times.10.sup.4, 1.2.times.10.sup.4, and 1.1.times.10.sup.4 members, respectively. The libraries were pooled for screening. Six rounds of screening by FACS were performed with decreasing concentrations of antigen, ranging from 500 nM to 160 nM of HAAH. An additional step of screening in the absence HAAH was performed to eliminate clones binding to the detection antibodies.

[0152] Ten clones were isolated and sequenced, and four of the clones were unique. The scFv display level (X-axis) and HAAH binding of these clones (Y-axis) is shown in FIGS. 22A-22D. The amino acid sequences of the CDRs of the wild type clone and the four mutagenized clones are shown in FIG. 23. Of the four unique sequences, three had a replacement in the region from FW1 to CDR3 with naive library sequences. The fourth clone had a replacement of the FW1 to CDR2 sequence with a naive library sequence. The dissociation constant of this mutant was 1 .mu.M-2 .mu.M, an order of magnitude improvement over the original wild type clone. Only two amino acid positions in the framework regions of this mutant differed from the germline framework regions.

[0153] The dissociation constants of various clones were determined. Binding vs. concentration of each clone is shown in FIG. 24. The K.sub.D measurements for each clone compared in the graph are as follows:

TABLE-US-00001 Wild type clone 6-22: >10 .mu.M Clone LLm13: 26 .+-. 8 nM Clone CM4: 17 to 5 nM Clone C4m18: 0.6 .+-. 0.1 nM

Example 8

Expression of Selected scFv Fragments as IgG Antibodies

[0154] Full length IgG antibodies were created from scFv fragments by inserting the isolated variable heavy and variable light chain sequences into an expression vector containing a sequence encoding the constant regions of the heavy and light IgG chains, the resulting vector expressing a full length IgG antibody (FIG. 25). In brief, the heavy and light chain sequences of the selected scFv fragments were separately amplified by PCR. The resulting fragments were then ligated into the expression vector pPNL501 at the appropriate locations (Pacific Northwest National Laboratory). The resulting vectors were cloned, isolated, and sequenced to confirm the correct sequences were inserted. The vectors were then transfected into COS-7L cells (Invitrogen) by standard methods. These resulting COS-7L cells express and secrete IgG into the culture medium. The medium was then isolated, and the IgG was purified using a Protein A resin by standard methods.

[0155] Binding specificity of the full length IgG antibodies was determined by ELISA and by binding to tumor cells expressing HAAH. For determination of binding specificity by ELISA, either HAAH or bovine serum albumin (BSA) was coated on an ELISA well, and different amounts of IgG were allowed to bind in each well. Detection was performed with a goat anti-human IgG-peroxidase conjugate and a chemoluminescent substrate. The results of this experiment for IgG 6-22, 6-23, 6-25, and 6-28 are shown in FIG. 26. For determination of binding specificity to HAAH-expressing tumor cells, different amounts of IgG antibodies were allowed to bind to H460 tumor cells, which express high levels of HAAH. Binding of IgG antibodies to the cells was detected using a goat anti-human IgG-FITC conjugate with detection by FACS. Results for IgG 6-22 and 6-23 are shown in FIG. 27; results for IgG LLm11, LLm13, LLm15, and CDRm4 are shown in FIG. 28.

[0156] The dissociation constants of IgG 6-22 and CDRm4 were determined. Different concentrations of IgG were allowed to bind either H460 or FOCUS cells at 4.degree. C. FOCUS cells are a hepatocellular carcinoma line which also expresses HAAH. After allowing IgG to bind, the cells were labeled with goat anti-human IgG-phycoerythrin conjugates and fluorescence was detected by FACS. Binding vs. concentration for 6-22 IgG is shown in FIG. 29; binding vs. concentration for CDRm4 IgG is shown in FIG. 30. The K.sub.D measurements determined are as follows:

TABLE-US-00002 H460 FOCUS 6-22 IgG 1.3 .+-. 0.2 nM 1.1 .+-. 0.2 nM CDRm4 IgG 1.0 .+-. 0.1 nM 0.7 .+-. 0.1 nM

[0157] Experiments were performed to determine if the anti-HAAH scFv fragments and derived IgG antibodies bind to similar epitopes on HAAH. FLAG-tagged scFv fragment 6-22 was allowed to bind H460 cells in the presence of 6-22 IgG or a control IgG Binding of the tagged scFv 6-22 was determined by labeling the FLAG tag, present only on the scFv 6-22, with a biotinylated mouse anti-FLAG antibody followed by binding to a streptavidin-phycoerythrin conjugate. IgG 6-22, but not control IgG, competed off the binding of scFv 6-22, indicating that the IgG form of 6-22 binds a similar epitope on HAAH as the scFv 6-22 (FIG. 31).

[0158] To further demonstrate that the scFv fragments and IgG antibodies bind to similar HAAH epitopes, yeast cells displaying different scFv fragments were allowed to bind 15 nM of HAAH in the presence of competing CDRm4 IgG, competing LLm11 IgG, or buffer alone. Binding of HAAH to cells displaying scFv fragments was detected by FACS as described above. The results, shown in FIG. 32, indicate that the IgG antibodies are capable of competing for binding to HAAH, further indicating that the IgG antibodies and scFv fragments bind similar epitopes.

Example 9

Second Generation Affinity Maturation of an Anti-HAAH scFv Fragment by Error-Prone PCR

[0159] Affinity maturation was performed on the CDRm4 mutant scFv to generate the second generation mutants C4m8 and C4m18. As described above, mutagenesis was performed by subjecting the CDRm4 scFv-encoding DNA to error prone PCR using nucleotide analogs. The mutagenized nucleic acids were re-expressed with yeast surface display and screened for binding using FACS as described above. DNA encoding binders was isolated, re-mutagenized, re-expressed and screened, and so forth for four rounds as described above. From this process, the improved second generation scFv clones were isolated.

[0160] The binding of improved clone C4m18 scFv mutant displayed on yeast was assayed for binding to HAAH as described above. This improved mutant was improved approximately 2-fold in both binding and display over CDRm4 (FIG. 33).

[0161] A number of embodiments of the invention have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.

Sequence CWU 1

1

3191125PRTArtificial SequenceSynthetically generated peptide 1Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 1252112PRTArtificial SequenceSynthetically generated peptide 2Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly 100 105 1103127PRTArtificial SequenceSynthetically generated peptide 3Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Val Ser Tyr Asp Gly Ser Gln Asp Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Lys Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr 100 105 110Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 1254109PRTArtificial SequenceSynthetically generated peptide 4Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser His Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Val Leu Ile 35 40 45Tyr Asp Val Ala Asn Arg Ala Ala Gly Thr Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Gln 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Ser Gly 100 1055124PRTArtificial SequenceSynthetically generated peptide 5Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asp 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ser Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Ala Gln Asn Asn Ile Ala Val Ala Gly Phe Asp 100 105 110Tyr Trp Gly Leu Gly Thr Leu Val Thr Val Ser Ser 115 1206110PRTArtificial SequenceSynthetically generated peptide 6Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Thr Leu Leu 35 40 45Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg65 70 75 80Ser Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Ser Gly Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Ser 100 105 1107123PRTArtificial SequenceSynthetically generated peptide 7Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Thr Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Asn Asp Gly Gly His Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asp Ser Met Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys 85 90 95Ala Lys Gly Arg Pro Trp Tyr Asp Pro Gly Ala Glu Tyr Phe Gln His 100 105 110Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 1208111PRTArtificial SequenceSynthetically generated peptide 8Gln Ser Ala Leu Ile Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln1 5 10 15Trp Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45Leu Ile Tyr Asp Val Ser Asp Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser 85 90 95Asn Thr Val Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ser 100 105 1109128PRTArtificial SequenceSynthetically generated peptide 9Gln Val Gln Leu Val Gln Ser Glu Gly Gly Val Val Arg Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Asp Tyr 20 25 30Ala Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ser Ile Asn Trp Asn Gly Gly Ser Thr Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Arg Val Ser Ser Gly Trp Pro Tyr Tyr Ser Leu Asp Val Trp Gly 100 105 110Gln Gly Thr Thr Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro Thr 115 120 12510112PRTArtificial SequenceSynthetically generated peptide 10Gln Ser Ala Leu Ile Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln1 5 10 15Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Ser Trp Tyr Gln His His Pro Gly Lys Ala Pro Gln Leu 35 40 45Met Ile Tyr Asp Val Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp 85 90 95Tyr Thr Tyr Ala Val Phe Gly Thr Gly Thr Gln Leu Thr Val Leu Ser 100 105 11011126PRTArtificial SequenceSynthetically generated peptide 11Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Ala Ile Ser Phe Asp Gly Asn Tyr Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Lys Val Asp Asp Thr Ala Val Tyr Phe Cys 85 90 95Ala Arg Asp Lys Ser Gly His Phe Gly Met Asp Val Trp Gly Gln Gly 100 105 110Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 12512107PRTArtificial SequenceSynthetically generated peptide 12Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Pro Ile 85 90 95Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 10513127PRTArtificial SequenceSynthetically generated peptide 13Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Pro Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Val Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Arg Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr 100 105 110Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12514107PRTArtificial SequenceSynthetically generated peptide 14Gly Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Pro Pro Arg1 5 10 15Glu Arg Ala Thr Leu Ser Cys Gly Thr Ser Gln Asn Val Ser His Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Arg Val Leu Thr 35 40 45Tyr Asp Val Ala Asn Arg Ala Ala Gly Thr Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Pro Ala Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Gln 85 90 95Ala Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10515127PRTArtificial SequenceSynthetically generated peptide 15Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Pro Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Ser Ser His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Arg Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr 100 105 110Asp Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12516113PRTArtificial SequenceSynthetically generated peptide 16Gly Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Pro Pro Arg1 5 10 15Glu Arg Val Thr Leu Ser Cys Gly Thr Ser Gln Asn Val Ser His His 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Arg Val Leu Thr 35 40 45Tyr Asp Val Ala Asn Lys Ala Ala Gly Thr Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Glu Thr Asp Phe Thr Pro Ala Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Gln 85 90 95Ala Phe Ser Pro Gly Thr Lys Val Asp Ile Lys Ser Gly Ser Glu Gln 100 105 110Lys17127PRTArtificial SequenceSynthetically generated peptide 17Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Pro Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Val Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Arg Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr His Tyr Tyr 100 105 110Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 12518107PRTArtificial SequenceSynthetically generated peptide 18Gly Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Pro Pro Arg1 5 10 15Glu Arg Ala Thr Leu Ser Cys Gly Thr Ser Gln Asn Val Ser His Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Arg Val Leu Thr 35 40 45Tyr Asp Val Ala Asn Arg Ala Ala Gly Thr Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Pro Ala Ile Ser Ser Leu Glu Leu65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Gln 85 90 95Ala Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10519125PRTArtificial SequenceSynthetically generated peptide 19Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Pro Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Arg Ala Ile Pro Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Phe Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Arg Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 12520111PRTArtificial SequenceSynthetically generated peptide 20Gln Ser Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser

Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 11021123PRTArtificial SequenceSynthetically generated peptide 21Gln Val Gln Leu Gln Gln Pro Gly Pro Gly Leu Val Glu Pro Ser Gln1 5 10 15Thr Leu Pro Leu Thr Arg Ala Ile Ser Gly Asp Ser Val Ser Gly Asn 20 25 30Gly Ala Ala Trp Ser Trp Ile Arg Gln Pro Pro Pro Arg Gly Leu Gly 35 40 45Trp Pro Gly Arg Thr Tyr Tyr Arg Pro Lys Arg Arg Asn Gly Tyr Ala 50 55 60Val Pro Ala Lys Ser Arg Met Thr Ile Ser Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly His Ser Ser Ser Trp Val Val Ser Phe 100 105 110Asp His Trp Gly Gln Gly Ala Thr Ile Ser Ser 115 12022112PRTArtificial SequenceSynthetically generated peptide 22Ser Gln Pro Val Pro Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly1 5 10 15Gln Arg Val Thr Val Ser Cys Ser Gly Ser Ser Ser Asn Ile Glu Ser 20 25 30Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Ala Ala Pro Arg Leu 35 40 45Phe Ile His Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Ser 50 55 60Ser Ser Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu65 70 75 80Gln Ser Lys Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser 85 90 95Leu Arg Ser Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 11023123PRTArtificial SequenceSynthetically generated peptide 23Gln Val Gln Leu Gln Arg Ser Gly Pro Gly Leu Val Lys Pro Pro Gln1 5 10 15Ala Pro Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile His Tyr Gly Ser Arg Trp Tyr Asn Asp Tyr Ala 50 55 60Ala Ser Ala Lys Ser Arg Val Thr Ile Asn Pro Asp Thr Pro Lys Gly65 70 75 80Gln Leu Pro Pro Gln Leu Ser Pro Val Thr Pro Lys Asp Ala Ala Ala 85 90 95Tyr Tyr Cys Ala Arg Thr Glu Cys Ser Ser Ser Trp Val Val Asn Phe 100 105 110Gly Tyr Trp Gly Gln Gly Ala Pro Val Thr Asp 115 12024111PRTArtificial SequenceSynthetically generated peptide 24Gln Pro Ala Leu Ile Gln Ser Pro Pro Val Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Pro Cys Ser Gly Ser Ser Phe Asn Ile Gly Ser Asn 20 25 30His Val Tyr Trp His Gln Gln Pro Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Val His Lys Ser Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Ser Pro 50 55 60Gly Pro Arg Pro Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Leu Gly Ala Gly Thr Lys Leu Thr Ala Arg Ser 100 105 11025125PRTArtificial SequenceSynthetically generated peptide 25Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Pro Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr His Arg Phe Lys Trp Tyr Asn Asp His Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Pro Val Thr Val Ser Ser 115 120 12526111PRTArtificial SequenceSynthetically generated peptide 26Gln Pro Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Arg Tyr Gln Gln Leu Pro Gly Ala Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Ser Ser Gly Val Pro Ala Arg Phe Ser 50 55 60Gly Pro Lys Ser Gly Ala Ala Ala Pro Leu Thr Thr Ser Gly Leu Gln65 70 75 80Ser Gly Asp Glu Ala Gly Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 11027125PRTArtificial SequenceSynthetically generated peptide 27Gln Val Gln Leu Gln Trp Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Arg Ala Ile Ser Gly Asn Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Ser Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Gly Ser Lys Arg Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Pro Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Ala Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Gly Ser Arg Val Val Asn Phe 100 105 110Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 12528109PRTArtificial SequenceSynthetically generated peptide 28Gln Pro Val Leu Thr Gln Ser Pro Pro Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val His Arg Tyr Gln Gln Pro Pro Gly Ala Ala Pro Glu Leu Leu 35 40 45Ile His Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Asp Ala Ile Ser Gly Pro Gln Ser Glu65 70 75 80Asp Glu Ala Asn Tyr Tyr Cys Val Ala Trp Asp Asp Ser Pro Cys Gly 85 90 95Tyr Val Phe Gly Ala Gly Thr Lys Leu Thr Val Leu Ser 100 10529125PRTArtificial SequenceSynthetically generated peptide 29Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Phe Ser Leu Thr Tyr Ala Val Ser Gly Gly Ser Ala Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Thr Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Gly Asp Ala Ala Val 85 90 95Cys Tyr Cys Ala Gly Ala Gly Tyr Ser Ser Ser Arg Ala Val Asn Phe 100 105 110Asp Cys Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 12530111PRTArtificial SequenceSynthetically generated peptide 30Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Pro Glu Gly Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Leu Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 11031264PRTArtificial SequenceSynthetically generated peptide 31Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro145 150 155 160Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu 180 185 190Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro 195 200 205Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala 210 215 220Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr225 230 235 240Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly 245 250 255Thr Lys Leu Thr Val Leu Ser Gly 26032256PRTArtificial SequenceSynthetically generated peptide 32Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Val Ser Tyr Asp Gly Ser Gln Asp Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Lys Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr 100 105 110Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 115 120 125Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu145 150 155 160Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 165 170 175Ser His Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 180 185 190Val Leu Ile Tyr Asp Val Ala Asn Arg Ala Ala Gly Thr Pro Ala Arg 195 200 205Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 210 215 220Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn225 230 235 240Trp Pro Gln Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Ser Gly 245 250 25533257PRTArtificial SequenceSynthetically generated peptide 33Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asp 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ser Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Ala Gln Asn Asn Ile Ala Val Ala Gly Phe Asp 100 105 110Tyr Trp Gly Leu Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly 115 120 125Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 130 135 140Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln145 150 155 160Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 165 170 175Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Thr Leu Leu 180 185 190Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 195 200 205Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 210 215 220Ser Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu225 230 235 240Ser Gly Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Ser Gly Ile 245 250 255Leu34257PRTArtificial SequenceSynthetically generated peptide 34Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Thr Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Asn Asp Gly Gly His Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asp Ser Met Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys 85 90 95Ala Lys Gly Arg Pro Trp Tyr Asp Pro Gly Ala Glu Tyr Phe Gln His 100 105 110Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly Ser 115 120 125Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln 130 135 140Ser Ala Leu Ile Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Trp145 150 155 160Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn 165 170 175Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Leu 180 185 190Ile Tyr Asp Val Ser Asp Arg Pro Ser Gly Val Ser Asn Arg Phe Ser 195 200 205Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln 210 215 220Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser Asn 225 230 235 240Thr Val Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ser Gly Ile 245 250 255Leu35263PRTArtificial SequenceSynthetically generated peptide 35Gln Val Gln Leu Val Gln Ser Glu Gly Gly Val Val Arg Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Asp Tyr 20 25 30Ala Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ser Ile Asn Trp Asn Gly Gly Ser Thr Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

85 90 95Ala Arg Val Ser Ser Gly Trp Pro Tyr Tyr Ser Leu Asp Val Trp Gly 100 105 110Gln Gly Thr Thr Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro Thr 115 120 125Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 130 135 140Gly Gly Gly Ser Gln Ser Ala Leu Ile Gln Pro Arg Ser Val Ser Gly145 150 155 160Ser Pro Gly Gln Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp 165 170 175Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr Gln His His Pro Gly Lys 180 185 190Ala Pro Gln Leu Met Ile Tyr Asp Val Thr Lys Arg Pro Ser Gly Val 195 200 205Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr 210 215 220Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser225 230 235 240Tyr Ala Gly Asp Tyr Thr Tyr Ala Val Phe Gly Thr Gly Thr Gln Leu 245 250 255Thr Val Leu Ser Gly Ile Leu 26036257PRTArtificial SequenceSynthetically generated peptide 36Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Ala Ile Ser Phe Asp Gly Asn Tyr Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Arg Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Lys Val Asp Asp Thr Ala Val Tyr Phe Cys 85 90 95Ala Arg Asp Lys Ser Gly His Phe Gly Met Asp Val Trp Gly Gln Gly 100 105 110Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Gly Ile 115 120 125Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140Gly Ser Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser145 150 155 160Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser 165 170 175Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 180 185 190Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg 195 200 205Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 210 215 220Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser225 230 235 240Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Ser Gly Ile 245 250 255Leu37256PRTArtificial SequenceSynthetically generated peptide 37Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Pro Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Val Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Arg Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr 100 105 110Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 115 120 125Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Asp Ser Gly Gly 130 135 140Gly Gly Ser Gly Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu145 150 155 160Pro Pro Arg Glu Arg Ala Thr Leu Ser Cys Gly Thr Ser Gln Asn Val 165 170 175Ser His Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Arg 180 185 190Val Leu Thr Tyr Asp Val Ala Asn Arg Ala Ala Gly Thr Pro Ala Arg 195 200 205Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Pro Ala Ile Ser Ser 210 215 220Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn225 230 235 240Trp Pro Gln Ala Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Ser Gly 245 250 25538264PRTArtificial SequenceSynthetically generated peptide 38Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Pro Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Ser Ser His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Arg Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr 100 105 110Asp Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 115 120 125Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Asp Ser Gly Gly 130 135 140Gly Gly Ser Gly Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu145 150 155 160Pro Pro Arg Glu Arg Val Thr Leu Ser Cys Gly Thr Ser Gln Asn Val 165 170 175Ser His His Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Arg 180 185 190Val Leu Thr Tyr Asp Val Ala Asn Lys Ala Ala Gly Thr Pro Ala Arg 195 200 205Phe Ser Gly Ser Gly Ser Glu Thr Asp Phe Thr Pro Ala Ile Ser Ser 210 215 220Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn225 230 235 240Trp Pro Gln Ala Phe Ser Pro Gly Thr Lys Val Asp Ile Lys Ser Gly 245 250 255Ser Glu Gln Lys Leu Ile Ser Glu 26039267PRTArtificial SequenceSynthetically generated peptide 39Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Pro Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Val Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Arg Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr His Tyr Tyr 100 105 110Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 115 120 125Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Asp Ser Gly Gly 130 135 140Gly Gly Ser Gly Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu145 150 155 160Pro Pro Arg Glu Arg Ala Thr Leu Ser Cys Gly Thr Ser Gln Asn Val 165 170 175Ser His Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Arg 180 185 190Val Leu Thr Tyr Asp Val Ala Asn Arg Ala Ala Gly Thr Pro Ala Arg 195 200 205Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Pro Ala Ile Ser Ser 210 215 220Leu Glu Leu Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn225 230 235 240Trp Pro Gln Ala Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Ser Gly 245 250 255Ser Glu Gln Lys Leu Ile Ser Glu Glu Ala Leu 260 26540266PRTArtificial SequenceSynthetically generated peptide 40Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Pro Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Arg Ala Ile Pro Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Phe Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Arg Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr Gly Ile Leu Gly Phe Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Ser Pro145 150 155 160Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu 180 185 190Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asp Asn Gln Arg Pro 195 200 205Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala 210 215 220Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr225 230 235 240Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly 245 250 255Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 260 26541264PRTArtificial SequenceSynthetically generated peptide 41Gln Val Gln Leu Gln Gln Pro Gly Pro Gly Leu Val Glu Pro Ser Gln1 5 10 15Thr Leu Pro Leu Thr Arg Ala Ile Ser Gly Asp Ser Val Ser Gly Asn 20 25 30Gly Ala Ala Trp Ser Trp Ile Arg Gln Pro Pro Pro Arg Gly Leu Gly 35 40 45Trp Pro Gly Arg Thr Tyr Tyr Arg Pro Lys Arg Arg Asn Gly Tyr Ala 50 55 60Val Pro Ala Lys Ser Arg Met Thr Ile Ser Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly His Ser Ser Ser Trp Val Val Ser Phe 100 105 110Asp His Trp Gly Gln Gly Ala Thr Ile Ser Ser Gly Asn Ala Ser Ala 115 120 125Pro Thr Gly Val Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 130 135 140Ser Gly Gly Gly Ser Ser Gln Pro Val Pro Thr Gln Ser Pro Ser Ala145 150 155 160Ser Gly Thr Pro Gly Gln Arg Val Thr Val Ser Cys Ser Gly Ser Ser 165 170 175Ser Asn Ile Glu Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly 180 185 190Ala Ala Pro Arg Leu Phe Ile His Lys Asn Asn Gln Arg Pro Ser Gly 195 200 205Val Pro Asp Arg Ser Ser Ser Ser Lys Ser Gly Thr Ala Ala Ser Leu 210 215 220Ala Ile Ser Gly Leu Gln Ser Lys Asp Glu Ala Asp Tyr Tyr Cys Ala225 230 235 240Ala Trp Asp Asp Ser Leu Arg Ser Tyr Val Phe Gly Thr Gly Thr Lys 245 250 255Leu Thr Val Leu Ser Gly Ile Leu 26042264PRTArtificial SequenceSynthetically generated peptide 42Gln Val Gln Leu Gln Arg Ser Gly Pro Gly Leu Val Lys Pro Pro Gln1 5 10 15Ala Pro Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile His Tyr Gly Ser Arg Trp Tyr Asn Asp Tyr Ala 50 55 60Ala Ser Ala Lys Ser Arg Val Thr Ile Asn Pro Asp Thr Pro Lys Gly65 70 75 80Gln Leu Pro Pro Gln Leu Ser Pro Val Thr Pro Lys Asp Ala Ala Ala 85 90 95Tyr Tyr Cys Ala Arg Thr Glu Cys Ser Ser Ser Trp Val Val Asn Phe 100 105 110Gly Tyr Trp Gly Gln Gly Ala Pro Val Thr Asp Gly Ser Ala Pro Ala 115 120 125Pro Thr Gly Ile Leu Gly Pro Gly Gly Gly Gly Ser Gly Gly Gly Gly 130 135 140Ser Gly Gly Gly Gly Pro Gln Pro Ala Leu Ile Gln Ser Pro Pro Val145 150 155 160Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Pro Cys Ser Gly Ser Ser 165 170 175Phe Asn Ile Gly Ser Asn His Val Tyr Trp His Gln Gln Pro Pro Gly 180 185 190Thr Ala Pro Lys Leu Leu Val His Lys Ser Asn Gln Arg Pro Ser Gly 195 200 205Val Pro Asp Arg Ser Pro Gly Pro Arg Pro Gly Thr Ala Ala Ser Leu 210 215 220Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala225 230 235 240Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Leu Gly Ala Gly Thr Lys 245 250 255Leu Thr Ala Arg Ser Gly Ile Leu 26043266PRTArtificial SequenceSynthetically generated peptide 43Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Pro Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr His Arg Phe Lys Trp Tyr Asn Asp His Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Pro Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr Gly Val Leu Gly Pro Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Pro Pro145 150 155 160Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Arg Tyr Gln Gln Leu 180 185 190Pro Gly Ala Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Ser 195 200 205Ser Gly Val Pro Ala Arg Phe Ser Gly Pro Lys Ser Gly Ala Ala Ala 210 215 220Pro Leu Thr Thr Ser Gly Leu Gln Ser Gly Asp Glu Ala Gly Tyr Tyr225 230 235 240Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly 245 250 255Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 260 26544264PRTArtificial SequenceSynthetically generated peptide 44Gln Val Gln Leu Gln Trp Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Arg Ala Ile Ser Gly Asn Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Ser Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Gly Ser Lys Arg Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Pro Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Ala Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Gly Ser Arg Val Val Asn Phe 100 105 110Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Arg Ala 115 120 125Pro Ala Pro Thr Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly

Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Pro Gln Pro Val Leu Thr Gln Ser Pro145 150 155 160Pro Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val His Arg Tyr Gln Gln Pro 180 185 190Pro Gly Ala Ala Pro Glu Leu Leu Ile His Lys Asn Asn Gln Arg Pro 195 200 205Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Asp 210 215 220Ala Ile Ser Gly Pro Gln Ser Glu Asp Glu Ala Asn Tyr Tyr Cys Val225 230 235 240Ala Trp Asp Asp Ser Pro Cys Gly Tyr Val Phe Gly Ala Gly Thr Lys 245 250 255Leu Thr Val Leu Ser Gly Ile Leu 26045265PRTArtificial SequenceSynthetically generated peptide 45Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Phe Ser Leu Thr Tyr Ala Val Ser Gly Gly Ser Ala Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Thr Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Gly Asp Ala Ala Val 85 90 95Cys Tyr Cys Ala Gly Ala Gly Tyr Ser Ser Ser Arg Ala Val Asn Phe 100 105 110Asp Cys Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Thr Pro Thr Gly Ile Leu Gly Ser Gly Gly Gly Ser Gly Gly Gly 130 135 140Gly Pro Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro Ser145 150 155 160Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser 165 170 175Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro 180 185 190Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser 195 200 205Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser 210 215 220Leu Ala Ile Ser Gly Leu Gln Pro Glu Gly Glu Ala Asp Tyr Tyr Cys225 230 235 240Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Leu Gly Thr Gly Thr 245 250 255Lys Leu Thr Val Leu Ser Gly Ile Leu 260 265467PRTArtificial SequenceSynthetically generated peptide 46Ser Asn Ser Ala Ala Trp Asn1 54718PRTArtificial SequenceSynthetically generated peptide 47Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser4813PRTArtificial SequenceSynthetically generated peptide 48Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe Asp Tyr1 5 104913PRTArtificial SequenceSynthetically generated peptide 49Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr1 5 105012PRTArtificial SequenceSynthetically generated peptide 50Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser1 5 105111PRTArtificial SequenceSynthetically generated peptide 51Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val1 5 10525PRTArtificial SequenceSynthetically generated peptide 52Ser Tyr Ala Met His1 55317PRTArtificial SequenceSynthetically generated peptide 53Val Val Ser Tyr Asp Gly Ser Gln Asp Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly5418PRTArtificial SequenceSynthetically generated peptide 54Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr Gly Met1 5 10 15Asp Val5511PRTArtificial SequenceSynthetically generated peptide 55Arg Ala Ser Gln Ser Val Ser His Tyr Leu Ala1 5 10567PRTArtificial SequenceSynthetically generated peptide 56Asp Val Ala Asn Arg Ala Ala1 5579PRTArtificial SequenceSynthetically generated peptide 57Gln Gln Arg Ser Asn Trp Pro Gln Thr1 5587PRTArtificial SequenceSynthetically generated peptide 58Ser Asp Ser Ala Ala Trp Asn1 559375PRTArtificial SequenceSynthetically generated peptide 59Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Pro Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly 100 105 110Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125Gly Gly Ser Arg Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr 130 135 140Pro Gly Gln Arg Val Thr Ile Pro Cys Ser Gly Ser Ser Ser Asn Ile145 150 155 160Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro 165 170 175Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp 180 185 190Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser 195 200 205Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp 210 215 220Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val225 230 235 240Leu Ser Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 245 250 255Ser Gly Gly Gly Gly Ser Arg Pro Val Leu Thr Gln Ser Pro Ser Ala 260 265 270Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Pro Cys Ser Gly Ser Ser 275 280 285Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly 290 295 300Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly305 310 315 320Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu 325 330 335Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala 340 345 350Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr Arg 355 360 365Leu Thr Val Leu Ser Gly Ser 370 3756012PRTArtificial SequenceSynthetically generated peptide 60Ala Gln Asn Asn Ile Ala Val Ala Gly Phe Asp Tyr1 5 1061256PRTArtificial SequenceSynthetically generated peptide 61Gln Val Gln Leu Val Glu Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Pro Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Val Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Tyr Cys 85 90 95Ala Arg Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr 100 105 110Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly 115 120 125Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Asp Ser Gly Gly 130 135 140Gly Gly Ser Gly Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu145 150 155 160Pro Pro Arg Glu Arg Ala Thr Leu Ser Cys Gly Thr Ser Gln Asn Val 165 170 175Ser His Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Arg 180 185 190Val Leu Thr Tyr Asp Val Ala Asn Arg Ala Ala Gly Thr Pro Ala Arg 195 200 205Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Pro Ala Ile Ser Ser 210 215 220Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn225 230 235 240Trp Pro Gln Ala Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Ser Gly 245 250 2556212PRTArtificial SequenceSynthetically generated peptide 62Thr Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser1 5 106310PRTArtificial SequenceSynthetically generated peptide 63Ala Ala Trp Asp Asp Ser Leu Ser Gly Val1 5 10645PRTArtificial SequenceSynthetically generated peptide 64Thr Tyr Ala Met His1 56517PRTArtificial SequenceSynthetically generated peptide 65Val Ile Ser Asn Asp Gly Gly His Lys Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly6614PRTArtificial SequenceSynthetically generated peptide 66Gly Arg Pro Trp Tyr Asp Pro Gly Ala Glu Tyr Phe Gln His1 5 106714PRTArtificial SequenceSynthetically generated peptide 67Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser1 5 10687PRTArtificial SequenceSynthetically generated peptide 68Asp Val Ser Asp Arg Pro Ser1 56910PRTArtificial SequenceSynthetically generated peptide 69Ser Ser Tyr Thr Ser Ser Asn Thr Val Leu1 5 10705PRTArtificial SequenceSynthetically generated peptide 70Asp Tyr Ala Met Gly1 57117PRTArtificial SequenceSynthetically generated peptide 71Ser Ile Asn Trp Asn Gly Gly Ser Thr Gly Tyr Ala Asp Ser Val Lys1 5 10 15Gly7212PRTArtificial SequenceSynthetically generated peptide 72Val Ser Ser Gly Trp Pro Tyr Tyr Ser Leu Asp Val1 5 107314PRTArtificial SequenceSynthetically generated peptide 73Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser1 5 10747PRTArtificial SequenceSynthetically generated peptide 74Asp Val Thr Lys Arg Pro Ser1 57511PRTArtificial SequenceSynthetically generated peptide 75Cys Ser Tyr Ala Gly Asp Tyr Thr Tyr Ala Val1 5 10765PRTArtificial SequenceSynthetically generated peptide 76Gly Tyr Ala Met His1 57717PRTArtificial SequenceSynthetically generated peptide 77Ala Ile Ser Phe Asp Gly Asn Tyr Lys Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly7810PRTArtificial SequenceSynthetically generated peptide 78Asp Lys Ser Gly His Phe Gly Met Asp Val1 5 107912PRTArtificial SequenceSynthetically generated peptide 79Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala1 5 10807PRTArtificial SequenceSynthetically generated peptide 80Gly Ala Ser Ser Arg Ala Thr1 5818PRTArtificial SequenceSynthetically generated peptide 81Gln Gln Tyr Gly Ser Pro Ile Thr1 5825PRTArtificial SequenceSynthetically generated peptide 82Ser His Ala Met His1 58317PRTArtificial SequenceSynthetically generated peptide 83Val Val Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val Lys1 5 10 15Gly8412PRTArtificial SequenceSynthetically generated peptide 84Cys Gly Thr Ser Gln Asn Val Ser His Tyr Leu Ala1 5 10859PRTArtificial SequenceSynthetically generated peptide 85Gln Gln Arg Ser Asn Trp Pro Gln Ala1 58617PRTArtificial SequenceSynthetically generated peptide 86Val Ile Ser His Asp Gly Ser Arg Asp Arg Tyr Ala Gly Ser Val Lys1 5 10 15Gly8718PRTArtificial SequenceSynthetically generated peptide 87Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr Tyr Tyr Tyr Asp Met1 5 10 15Asp Val8811PRTArtificial SequenceSynthetically generated peptide 88Gly Thr Ser Gln Asn Val Ser His His Leu Ala1 5 10897PRTArtificial SequenceSynthetically generated peptide 89Asp Val Ala Asn Lys Ala Ala1 5909PRTArtificial SequenceSynthetically generated peptide 90Gln Gln Arg Ser Asn Trp Pro Gln Ala1 59118PRTArtificial SequenceSynthetically generated peptide 91Val Gly Arg Ser Ser Asn Trp Phe Ser Arg Tyr His Tyr Tyr Gly Met1 5 10 15Asp Val9211PRTArtificial SequenceSynthetically generated peptide 92Gly Thr Ser Gln Asn Val Ser His Tyr Leu Ala1 5 10939PRTArtificial SequenceSynthetically generated peptide 93Gln Gln Arg Ser Asn Trp Pro Gln Ala1 5948PRTArtificial SequenceSynthetically generated peptide 94Ser Ser Asn Ser Ala Ala Trp Asn1 59518PRTArtificial SequenceSynthetically generated peptide 95Arg Thr Tyr Tyr Arg Ser Lys Arg Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser9613PRTArtificial SequenceSynthetically generated peptide 96Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe Asp Tyr1 5 109719PRTArtificial SequenceSynthetically generated peptide 97Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg9812PRTArtificial SequenceSynthetically generated peptide 98Lys Leu Leu Ile Tyr Lys Asp Asn Gln Arg Pro Ser1 5 10997PRTArtificial SequenceSynthetically generated peptide 99Gly Asn Gly Ala Ala Trp Ser1 510018PRTArtificial SequenceSynthetically generated peptide 100Arg Thr Tyr Tyr Arg Pro Lys Arg Arg Asn Gly Tyr Ala Val Pro Ala1 5 10 15Lys Ser10113PRTArtificial SequenceSynthetically generated peptide 101Thr Gly His Ser Ser Ser Trp Val Val Ser Phe Asp His1 5 1010213PRTArtificial SequenceSynthetically generated peptide 102Ser Gly Ser Ser Ser Asn Ile Glu Ser Asn Tyr Val Tyr1 5 1010312PRTArtificial SequenceSynthetically generated peptide 103Arg Leu Phe Ile His Lys Asn Asn Gln Arg Pro Ser1 5 1010411PRTArtificial SequenceSynthetically generated peptide 104Ala Ala Trp Asp Asp Ser Leu Arg Ser Tyr Val1 5 1010518PRTArtificial SequenceSynthetically generated peptide 105Arg Ile His Tyr Gly Ser Arg Trp Tyr Asn Asp Tyr Ala Ala Ser Ala1 5 10 15Lys Ser10613PRTArtificial SequenceSynthetically generated peptide 106Thr Glu Cys Ser Ser Ser Trp Val Val Asn Phe Gly Tyr1 5 1010713PRTArtificial SequenceSynthetically generated peptide 107Ser Gly Ser Ser Phe Asn Ile Gly Ser Asn His Val Tyr1 5 1010812PRTArtificial SequenceSynthetically generated peptide 108Lys Leu Leu Val His Lys Ser Asn Gln Arg Pro Ser1 5 1010918PRTArtificial SequenceSynthetically generated peptide 109Arg Thr Tyr His Arg Phe Lys Trp Tyr Asn Asp His Ala Val Ser Val1 5 10 15Lys Ser11012PRTArtificial SequenceSynthetically generated peptide 110Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Ser Ser1 5 101117PRTArtificial SequenceSynthetically generated peptide 111Ser Asn Ser Ala Ala Trp Ser1 511218PRTArtificial SequenceSynthetically generated peptide 112Arg Thr Tyr Tyr Gly Ser Lys Arg Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser11313PRTArtificial SequenceSynthetically generated peptide 113Thr Gly Tyr Ser Gly Ser Arg Val Val Asn Phe Gly Tyr1 5 1011413PRTArtificial SequenceSynthetically generated peptide 114Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val His1 5 1011512PRTArtificial SequenceSynthetically generated peptide 115Glu Leu Leu Ile His Lys Asn Asn Gln Arg Pro Ser1 5 1011611PRTArtificial SequenceSynthetically generated peptide 116Val Ala Trp Asp Asp Ser Pro Cys Gly Tyr Val1 5 1011713PRTArtificial SequenceSynthetically generated peptide 117Ala Gly Tyr Ser Ser Ser Arg Ala Val Asn Phe Asp Cys1 5 1011813PRTArtificial SequenceSynthetically generated peptide 118Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe Asp Tyr1 5

10119131PRTArtificial SequenceSynthetically generated peptide 119Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Ile 20 25 30Thr Ala Ala Trp Asn Trp Leu Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr His Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Val Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu His Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Arg Leu Ala Arg Gly Gly Pro Ser Ala His Ala Phe Glu 100 105 110Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser 130120111PRTArtificial SequenceSynthetically generated peptide 120Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110121120PRTArtificial SequenceSynthetically generated peptide 121Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Val Ile Ser Gly Asp Ser Val Ser Asn Asn 20 25 30Ser Ala Val Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ile Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Arg Thr Gly Ala Gly Val Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser 115 120122111PRTArtificial SequenceSynthetically generated peptide 122Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110123124PRTArtificial SequenceSynthetically generated peptide 123Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Gly Val Ser Gly Asn 20 25 30Asn Val Ile Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Leu Leu 50 55 60Pro Ser Val Lys Ser Arg Ile Ala Ile Asn Pro Asp Thr Ser Lys Ser65 70 75 80Gln Phe Ser Leu Gln Leu Ser Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Arg Ala Val Ala Gly Asn Gln Tyr Phe Asp 100 105 110Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser 115 120124111PRTArtificial SequenceSynthetically generated peptide 124Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110125129PRTArtificial SequenceSynthetically generated peptide 125Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Tyr 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Ser Leu Ala Ala Ala Ala Gly Thr Val Asp Tyr Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro 115 120 125Thr126111PRTArtificial SequenceSynthetically generated peptide 126Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110127126PRTArtificial SequenceSynthetically generated peptide 127Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Leu Ser Val Lys Ser Arg Ile Asn Ile Asn Ala Asp Thr Ser Lys Ser65 70 75 80Gln Phe Ser Leu Gln Leu Asp Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Phe Cys Ala Lys Asp Arg Leu Leu Tyr Asn Tyr Gly Ser Asn Ala 100 105 110Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125128111PRTArtificial SequenceSynthetically generated peptide 128Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110129136PRTArtificial SequenceSynthetically generated peptide 129Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Asp Thr Pro Arg Tyr Cys Ser Gly Gly Ser Cys 100 105 110Tyr Lys Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser 115 120 125Ser Ala Ser Thr Lys Ser Pro Ser 130 135130111PRTArtificial SequenceSynthetically generated peptide 130Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110131124PRTArtificial SequenceSynthetically generated peptide 131Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Ser Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys 20 25 30Gly Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ala Tyr Tyr Trp Ser Lys Trp Tyr Tyr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Leu Thr Pro Glu Asp Thr Ala Val 85 90 95Phe Tyr Cys Arg Ala Thr Ser Thr Tyr Tyr Leu Pro Gly Gly Leu Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120132111PRTArtificial SequenceSynthetically generated peptide 132Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110133121PRTArtificial SequenceSynthetically generated peptide 133Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Thr Ile Ser Gly Asp Ser Val Ser Ala Asp 20 25 30Arg Val Ala Trp Asn Trp Ile Arg Gln Ser Pro Leu Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Phe Tyr Arg Ser Lys Trp Met Val Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ser Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Met 85 90 95Tyr Tyr Cys Ala Arg Ala Thr Thr Arg Gly Tyr Phe Asp Leu Trp Gly 100 105 110Arg Gly Thr Leu Val Thr Val Ser Ser 115 120134111PRTArtificial SequenceSynthetically generated peptide 134Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110135123PRTArtificial SequenceSynthetically generated peptide 135Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Val Val Ser Gly Asp Gly Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Met Lys Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asp Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Arg Ser Gly Arg Thr Gly Gly Tyr Phe Asp Leu 100 105 110Trp Gly Arg Gly Thr Leu Val Ala Val Ser Ser 115 120136111PRTArtificial SequenceSynthetically generated peptide 136Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110137120PRTArtificial SequenceSynthetically generated peptide 137Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Pro Gly His Ser Val Gly Ser Ser 20 25 30Asn Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Phe Tyr Gly Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Leu Lys Ser Arg Leu Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Arg Thr Gly Thr Gly Ile Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser 115 120138111PRTArtificial SequenceSynthetically generated peptide 138Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu

Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110139118PRTArtificial SequenceSynthetically generated peptide 139Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly His Ser Val Gly Ser Ser 20 25 30Asn Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Phe Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Thr Arg Ile Ser Ile Asn Pro Asp Thr Ala Lys Asn65 70 75 80Gln Phe Ser Leu His Leu Asn Ser Val Thr Ala Glu Asp Thr Gly Val 85 90 95Tyr Tyr Cys Arg Gln Gln Lys Arg Leu Asp Ser Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 115140111PRTArtificial SequenceSynthetically generated peptide 140Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110141132PRTArtificial SequenceSynthetically generated peptide 141Pro Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Pro Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Asp Asp Thr Ala Ile 85 90 95Tyr Tyr Cys Ala Arg Ala Ser Asp Tyr Gly Asp Tyr Phe Tyr Tyr Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr 130142111PRTArtificial SequenceSynthetically generated peptide 142Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110143125PRTArtificial SequenceSynthetically generated peptide 143Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ala Ile Lys Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Arg Ala Gly Arg Ser Phe Asp Leu Trp Gly Arg Gly Thr 100 105 110Leu Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro Thr 115 120 125144111PRTArtificial SequenceSynthetically generated peptide 144Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110145128PRTArtificial SequenceSynthetically generated peptide 145Gln Val Gln Leu Gln Gln Ser Ser Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Val Ser Gly Asn 20 25 30Ser Gly Val Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Tyr Thr Tyr Lys Trp Tyr Ile Asp Tyr 50 55 60Ala Val Ser Val Lys Ser Arg Ile Thr Val Asn Pro Asp Thr Ser Arg65 70 75 80Asn Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala 85 90 95Val Tyr Tyr Cys Ala Arg Val Asp Tyr Thr Gly Ser Pro Val Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro Thr 115 120 125146111PRTArtificial SequenceSynthetically generated peptide 146Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110147132PRTArtificial SequenceSynthetically generated peptide 147Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Arg Gly Arg Ile Thr Ile Asn Ala Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr 130148111PRTArtificial SequenceSynthetically generated peptide 148Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 100 105 110149265PRTArtificial SequenceSynthetically generated peptide 149Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Ile 20 25 30Thr Ala Ala Trp Asn Trp Leu Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr His Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Val Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu His Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Arg Leu Ala Arg Gly Gly Pro Ser Ala His Ala Phe Glu 100 105 110Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro Ser145 150 155 160Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser 165 170 175 Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro 180 185 190Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser 195 200 205Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser 210 215 220Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys225 230 235 240Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr 245 250 255Lys Leu Thr Val Leu Ser Gly Ile Leu 260 265150254PRTArtificial SequenceSynthetically generated peptide 150Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Val Ile Ser Gly Asp Ser Val Ser Asn Asn 20 25 30Ser Ala Val Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ile Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Arg Thr Gly Ala Gly Val Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly Ser Gly Gly Gly 115 120 125Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu 130 135 140Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile145 150 155 160Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp 165 170 175Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn 180 185 190Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser 195 200 205Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu 210 215 220Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val225 230 235 240Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 245 250151258PRTArtificial SequenceSynthetically generated peptide 151Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Gly Val Ser Gly Asn 20 25 30Asn Val Ile Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Leu Leu 50 55 60Pro Ser Val Lys Ser Arg Ile Ala Ile Asn Pro Asp Thr Ser Lys Ser65 70 75 80Gln Phe Ser Leu Gln Leu Ser Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Arg Ala Val Ala Gly Asn Gln Tyr Phe Asp 100 105 110Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly 115 120 125Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 130 135 140Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln145 150 155 160Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 165 170 175Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 180 185 190Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 195 200 205Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln 210 215 220Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu225 230 235 240Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly 245 250 255Ile Leu152263PRTArtificial SequenceSynthetically generated peptide 152Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Tyr 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Ser Leu Ala Ala Ala Ala Gly Thr Val Asp Tyr Trp 100 105 110Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro 115 120 125Thr Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 130 135 140Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser145 150 155 160Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser 165 170 175Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr 180 185 190Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val 195 200 205Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala 210 215 220Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala225 230 235 240Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu 245 250 255Thr Val Leu Ser Gly Ile Leu 260153260PRTArtificial SequenceSynthetically generated peptide 153Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys

Trp Tyr Asn Asp Tyr Ala 50 55 60Leu Ser Val Lys Ser Arg Ile Asn Ile Asn Ala Asp Thr Ser Lys Ser65 70 75 80Gln Phe Ser Leu Gln Leu Asp Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Phe Cys Ala Lys Asp Arg Leu Leu Tyr Asn Tyr Gly Ser Asn Ala 100 105 110Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Ile 115 120 125Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140Gly Ser Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro145 150 155 160Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly 165 170 175Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys 180 185 190Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg 195 200 205Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly 210 215 220Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp225 230 235 240Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu 245 250 255Ser Gly Ile Leu 260154270PRTArtificial SequenceSynthetically generated peptide 154Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Asp Thr Pro Arg Tyr Cys Ser Gly Gly Ser Cys 100 105 110Tyr Lys Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser 115 120 125Ser Ala Ser Thr Lys Ser Pro Ser Gly Ile Leu Gly Ser Gly Gly Gly 130 135 140Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu145 150 155 160Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile 165 170 175Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp 180 185 190Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn 195 200 205Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser 210 215 220Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu225 230 235 240Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val 245 250 255Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 260 265 270155258PRTArtificial SequenceSynthetically generated peptide 155Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Ser Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys 20 25 30Gly Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ala Tyr Tyr Trp Ser Lys Trp Tyr Tyr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Leu Thr Pro Glu Asp Thr Ala Val 85 90 95Phe Tyr Cys Arg Ala Thr Ser Thr Tyr Tyr Leu Pro Gly Gly Leu Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Ile Leu Gly 115 120 125Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 130 135 140Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln145 150 155 160Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 165 170 175Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 180 185 190Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 195 200 205Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln 210 215 220Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu225 230 235 240Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly 245 250 255Ile Leu156255PRTArtificial SequenceSynthetically generated peptide 156Gln Val Gln Leu Gln Gln Ser Gly Ala Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Thr Ile Ser Gly Asp Ser Val Ser Ala Asp 20 25 30Arg Val Ala Trp Asn Trp Ile Arg Gln Ser Pro Leu Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Phe Tyr Arg Ser Lys Trp Met Val Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ser Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Met 85 90 95Tyr Tyr Cys Ala Arg Ala Thr Thr Arg Gly Tyr Phe Asp Leu Trp Gly 100 105 110Arg Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly Ser Gly Gly 115 120 125Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val 130 135 140Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr145 150 155 160Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr 165 170 175Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys 180 185 190Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys 195 200 205Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp 210 215 220Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr225 230 235 240Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 245 250 255157257PRTArtificial SequenceSynthetically generated peptide 157Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Val Val Ser Gly Asp Gly Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Met Lys Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asp Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Ala Arg Arg Ser Gly Arg Thr Gly Gly Tyr Phe Asp Leu 100 105 110Trp Gly Arg Gly Thr Leu Val Ala Val Ser Ser Gly Ile Leu Gly Ser 115 120 125Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln 130 135 140Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg145 150 155 160Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr 165 170 175Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 180 185 190Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 195 200 205Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser 210 215 220Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Arg225 230 235 240Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly Ile 245 250 255Leu158254PRTArtificial SequenceSynthetically generated peptide 158Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Pro Gly His Ser Val Gly Ser Ser 20 25 30Asn Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Phe Tyr Gly Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Leu Lys Ser Arg Leu Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Arg Thr Gly Thr Gly Ile Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly Ser Gly Gly Gly 115 120 125Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu 130 135 140Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile145 150 155 160Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp 165 170 175Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn 180 185 190Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser 195 200 205Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu 210 215 220Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val225 230 235 240Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 245 250159252PRTArtificial SequenceSynthetically generated peptide 159Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly His Ser Val Gly Ser Ser 20 25 30Asn Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Phe Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Thr Arg Ile Ser Ile Asn Pro Asp Thr Ala Lys Asn65 70 75 80Gln Phe Ser Leu His Leu Asn Ser Val Thr Ala Glu Asp Thr Gly Val 85 90 95Tyr Tyr Cys Arg Gln Gln Lys Arg Leu Asp Ser Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser 115 120 125Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln 130 135 140Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys145 150 155 160Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln 165 170 175Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln 180 185 190Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr 195 200 205Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp 210 215 220Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly225 230 235 240Thr Gly Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 245 250160266PRTArtificial SequenceSynthetically generated peptide 160Pro Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Pro Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Asp Asp Thr Ala Ile 85 90 95Tyr Tyr Cys Ala Arg Ala Ser Asp Tyr Gly Asp Tyr Phe Tyr Tyr Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro145 150 155 160Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu 180 185 190Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro 195 200 205Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala 210 215 220Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr225 230 235 240Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly 245 250 255Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 260 265161259PRTArtificial SequenceSynthetically generated peptide 161Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Ala Ile Lys Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Arg Ala Gly Arg Ser Phe Asp Leu Trp Gly Arg Gly Thr 100 105 110Leu Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro Thr Gly Ile Leu 115 120 125Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 130 135 140Ser Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly145 150 155 160Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser 165 170 175Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 180 185 190Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe 195 200 205Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu 210 215 220Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser225 230 235 240Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu Ser 245 250 255Gly Ile Leu162262PRTArtificial SequenceSynthetically generated peptide 162Gln Val Gln Leu Gln Gln Ser Ser Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Val Ser Gly Asn 20 25 30Ser Gly Val Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Tyr Thr Tyr Lys Trp Tyr Ile Asp Tyr 50 55 60Ala Val Ser Val Lys Ser Arg Ile Thr Val Asn Pro Asp Thr Ser Arg65 70 75 80Asn Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala 85 90 95Val Tyr Tyr Cys Ala Arg Val Asp Tyr Thr Gly Ser Pro Val Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala Ser Ala Pro Thr 115

120 125Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 130 135 140Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly145 150 155 160Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn 165 170 175Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala 180 185 190Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro 195 200 205Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile 210 215 220Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp225 230 235 240Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr 245 250 255Val Leu Ser Gly Ile Leu 260163266PRTArtificial SequenceSynthetically generated peptide 163Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Arg Gly Arg Ile Thr Ile Asn Ala Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe 100 105 110Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro145 150 155 160Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu 180 185 190Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro 195 200 205Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala 210 215 220Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr225 230 235 240Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly 245 250 255Thr Lys Leu Thr Val Leu Ser Gly Ile Leu 260 2651647PRTArtificial SequenceSynthetically generated peptide 164Ser Ile Thr Ala Ala Trp Asn1 516518PRTArtificial SequenceSynthetically generated peptide 165Arg Thr Tyr His Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser16616PRTArtificial SequenceSynthetically generated peptide 166Gly Leu Ala Ala Arg Gly Gly Gly Pro Ser Ala His Ala Phe Glu Ile1 5 10 151678PRTArtificial SequenceSynthetically generated peptide 167Ser Ser Ile Thr Ala Ala Trp Asn1 516819PRTArtificial SequenceSynthetically generated peptide 168Arg Thr Tyr His Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg16916PRTArtificial SequenceSynthetically generated peptide 169Gly Leu Ala Ala Arg Gly Gly Gly Pro Ser Ala His Ala Phe Glu Ile1 5 10 151707PRTArtificial SequenceSynthetically generated peptide 170Asn Asn Ser Ala Val Trp Asn1 51718PRTArtificial SequenceSynthetically generated peptide 171Ser Asn Asn Ser Ala Val Trp Asn1 51728PRTArtificial SequenceSynthetically generated peptide 172Arg Thr Gly Ala Gly Val Asp Tyr1 51738PRTArtificial SequenceSynthetically generated peptide 173Ser Gly Asn Asn Val Ile Trp Asn1 517419PRTArtificial SequenceSynthetically generated peptide 174Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Leu Leu Pro Ser Val1 5 10 15Lys Ser Arg17512PRTArtificial SequenceSynthetically generated peptide 175Thr Arg Ala Val Ala Gly Asn Gln Tyr Phe Asp Leu1 5 101767PRTArtificial SequenceSynthetically generated peptide 176Gly Asn Asn Val Ile Trp Asn1 517718PRTArtificial SequenceSynthetically generated peptide 177Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Leu Leu Pro Ser Val1 5 10 15Lys Ser17812PRTArtificial SequenceSynthetically generated peptide 178Thr Arg Ala Val Ala Gly Asn Gln Tyr Phe Asp Leu1 5 101798PRTArtificial SequenceSynthetically generated peptide 179Ser Ser Tyr Ser Ala Ala Trp Asn1 518019PRTArtificial SequenceSynthetically generated peptide 180Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg18110PRTArtificial SequenceSynthetically generated peptide 181Leu Ala Ala Ala Ala Gly Thr Val Asp Tyr1 5 101827PRTArtificial SequenceSynthetically generated peptide 182Ser Tyr Ser Ala Ala Trp Asn1 51838PRTArtificial SequenceSynthetically generated peptide 183Ser Ser Asn Ser Ala Ala Trp Asn1 518410PRTArtificial SequenceSynthetically generated peptide 184Leu Ala Ala Ala Ala Gly Thr Val Asp Tyr1 5 1018519PRTArtificial SequenceSynthetically generated peptide 185Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Leu Ser Val1 5 10 15Lys Ser Arg18613PRTArtificial SequenceSynthetically generated peptide 186Asp Arg Leu Leu Tyr Asn Tyr Gly Ser Asn Ala Met Asp1 5 101878PRTArtificial SequenceSynthetically generated peptide 187Ser Ser Asn Ser Ala Ala Trp Asn1 518819PRTArtificial SequenceSynthetically generated peptide 188Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg18918PRTArtificial SequenceSynthetically generated peptide 189Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Leu Ser Val1 5 10 15Lys Ser19014PRTArtificial SequenceSynthetically generated peptide 190Asp Arg Leu Leu Tyr Asn Tyr Gly Ser Asn Ala Met Asp Val1 5 1019116PRTArtificial SequenceSynthetically generated peptide 191Asp Thr Pro Arg Tyr Cys Ser Gly Gly Ser Cys Tyr Lys Tyr Phe Asp1 5 10 151928PRTArtificial SequenceSynthetically generated peptide 192Ser Ser Lys Gly Ala Ala Trp Asn1 519319PRTArtificial SequenceSynthetically generated peptide 193Arg Ala Tyr Tyr Trp Ser Lys Trp Tyr Tyr Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg19414PRTArtificial SequenceSynthetically generated peptide 194Gly Ala Thr Ser Thr Tyr Tyr Leu Pro Gly Gly Leu Asp Val1 5 101958PRTArtificial SequenceSynthetically generated peptide 195Ser Ala Asp Arg Val Ala Trp Asn1 519617PRTArtificial SequenceSynthetically generated peptide 196Asp Thr Pro Arg Tyr Cys Ser Gly Gly Ser Cys Tyr Lys Tyr Phe Asp1 5 10 15Tyr19719PRTArtificial SequenceSynthetically generated peptide 197Arg Ile Phe Tyr Arg Ser Lys Trp Met Val Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg1989PRTArtificial SequenceSynthetically generated peptide 198Ala Thr Thr Arg Gly Tyr Phe Asp Leu1 519919PRTArtificial SequenceSynthetically generated peptide 199Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Met1 5 10 15Lys Gly Arg2007PRTArtificial SequenceSynthetically generated peptide 200Ser Lys Gly Ala Ala Trp Asn1 520118PRTArtificial SequenceSynthetically generated peptide 201Arg Ala Tyr Tyr Trp Ser Lys Trp Tyr Tyr Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser20214PRTArtificial SequenceSynthetically generated peptide 202Gly Ala Thr Ser Thr Tyr Tyr Leu Pro Gly Gly Leu Asp Val1 5 1020311PRTArtificial SequenceSynthetically generated peptide 203Arg Ser Gly Arg Thr Gly Gly Tyr Phe Asp Leu1 5 102048PRTArtificial SequenceSynthetically generated peptide 204Gly Ser Ser Asn Ala Ala Trp Asn1 520519PRTArtificial SequenceSynthetically generated peptide 205Arg Ile Phe Tyr Gly Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Leu1 5 10 15Lys Ser Arg2067PRTArtificial SequenceSynthetically generated peptide 206Ala Asp Arg Val Ala Trp Asn1 520718PRTArtificial SequenceSynthetically generated peptide 207Arg Ile Phe Tyr Arg Ser Lys Trp Met Val Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser2089PRTArtificial SequenceSynthetically generated peptide 208Ala Thr Thr Arg Gly Tyr Phe Asp Leu1 52098PRTArtificial SequenceSynthetically generated peptide 209Arg Thr Gly Thr Gly Ile Asp Tyr1 521019PRTArtificial SequenceSynthetically generated peptide 210Arg Ile Phe Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Thr Arg2118PRTArtificial SequenceSynthetically generated peptide 211Gly Gln Gln Lys Arg Leu Asp Ser1 521213PRTArtificial SequenceSynthetically generated peptide 212Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe Asp Tyr1 5 1021318PRTArtificial SequenceSynthetically generated peptide 213Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Met1 5 10 15Lys Gly21411PRTArtificial SequenceSynthetically generated peptide 214Arg Ser Gly Arg Thr Gly Gly Tyr Phe Asp Leu1 5 1021513PRTArtificial SequenceSynthetically generated peptide 215Ala Ser Asp Tyr Gly Asp Tyr Phe Tyr Tyr Phe Asp Tyr1 5 1021619PRTArtificial SequenceSynthetically generated peptide 216Arg Ile Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg2178PRTArtificial SequenceSynthetically generated peptide 217Gly Ala Gly Arg Ser Phe Asp Leu1 52187PRTArtificial SequenceSynthetically generated peptide 218Ser Ser Asn Ala Ala Trp Asn1 521918PRTArtificial SequenceSynthetically generated peptide 219Arg Ile Phe Tyr Gly Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Leu1 5 10 15Lys Ser2208PRTArtificial SequenceSynthetically generated peptide 220Arg Thr Gly Thr Gly Ile Asp Tyr1 52218PRTArtificial SequenceSynthetically generated peptide 221Ser Gly Asn Ser Gly Val Trp Asn1 522220PRTArtificial SequenceSynthetically generated peptide 222Arg Thr Tyr Tyr Tyr Thr Tyr Lys Trp Tyr Ile Asp Tyr Ala Val Ser1 5 10 15Val Lys Ser Arg 202238PRTArtificial SequenceSynthetically generated peptide 223Val Asp Tyr Thr Gly Ser Pro Val1 52247PRTArtificial SequenceSynthetically generated peptide 224Ser Ser Asn Ala Ala Trp Asn1 522518PRTArtificial SequenceSynthetically generated peptide 225Arg Ile Phe Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Thr2268PRTArtificial SequenceSynthetically generated peptide 226Gly Gln Gln Lys Arg Leu Asp Ser1 522719PRTArtificial SequenceSynthetically generated peptide 227Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Arg Gly Arg22813PRTArtificial SequenceSynthetically generated peptide 228Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Phe Asp Tyr1 5 1022910PRTArtificial SequenceSynthetically generated peptide 229Ser Gln Xaa Val Ser Xaa Xaa Xaa Leu Ala1 5 102307PRTArtificial SequenceSynthetically generated peptide 230Asp Val Ala Asn Xaa Ala Ala1 52318PRTArtificial SequenceSynthetically generated peptide 231Gln Gln Arg Ser Gln Trp Pro Gln1 523213PRTArtificial SequenceSynthetically generated peptide 232Ala Ser Asp Tyr Gly Asp Tyr Phe Tyr Tyr Phe Asp Tyr1 5 102334PRTArtificial SequenceSynthetically generated peptide 233Xaa Ala Met Xaa12347PRTArtificial SequenceSynthetically generated peptide 234Tyr Ala Xaa Ser Val Lys Xaa1 523513PRTArtificial SequenceSynthetically generated peptide 235Ser Gly Ser Ser Ser Asn Ile Xaa Ser Asn Xaa Val Xaa1 5 102367PRTArtificial SequenceSynthetically generated peptide 236Xaa Xaa Xaa Ala Ala Trp Xaa1 523718PRTArtificial SequenceSynthetically generated peptide 237Arg Ile Tyr Tyr Arg Ser Lys Trp Tyr Tyr Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser2388PRTArtificial SequenceSynthetically generated peptide 238Gly Ala Gly Arg Ser Phe Asp Leu1 523918PRTArtificial SequenceSynthetically generated peptide 239Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Ala Xaa Xaa Xaa1 5 10 15Lys Ser2407PRTArtificial SequenceSynthetically generated peptide 240Asp Val Xaa Xaa Arg Pro Ser1 524111PRTArtificial SequenceSynthetically generated peptide 241Leu Xaa Xaa Xaa Xaa Xaa Asn Gln Arg Pro Ser1 5 102428PRTArtificial SequenceSynthetically generated peptide 242Gly Asn Ser Gly Val Trp Asn Trp1 524318PRTArtificial SequenceSynthetically generated peptide 243Thr Tyr Tyr Tyr Thr Tyr Lys Trp Tyr Ile Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser2448PRTArtificial SequenceSynthetically generated peptide 244Val Asp Tyr Thr Gly Ser Pro Val1 52455PRTArtificial SequenceSynthetically generated peptide 245Ala Trp Asp Asp Ser1 52469PRTArtificial SequenceSynthetically generated peptide 246Ser Ser Ser Trp Val Val Xaa Phe Xaa1 524729PRTArtificial SequenceSynthetically generated peptide 247Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Cys Xaa Xaa Xaa Xaa Cys Xaa Xaa1 5 10 15Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Cys Xaa Cys Xaa Cys 20 2524819PRTArtificial SequenceSynthetically generated peptide 248Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg2498PRTArtificial SequenceSynthetically generated peptide 249Ser Ser Asn Ser Ala Ala Trp Asn1 52508PRTArtificial SequenceSynthetically generated peptide 250Gly Ser Ser Asn Ala Ala Trp Asn1 525119PRTArtificial SequenceSynthetically generated peptide 251Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser Arg2528PRTArtificial SequenceSynthetically generated peptide 252Ser Ser Asn Ser Ala Ala Trp Asn1 52538PRTArtificial SequenceSynthetically generated peptide 253Ser Ser Asn Ser Ala Ala Trp Asn1 5254121PRTArtificial SequenceSynthetically generated peptide 254Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Thr Lys Trp Tyr Asn Glu Tyr Ala 50 55 60Ala Ser Val Lys Gly Arg Ala Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Thr Asp Pro Lys Gly Val Thr Thr Gln Tyr Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val Ser Ser 115 120255110PRTArtificial SequenceSynthetically generated peptide 255Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Thr Ser Asn Ile Gly Arg Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Ala Pro Ala Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Thr Ser Leu Ala Ile Ser Gly Leu Arg65 70 75 80Ser

Glu Asp Glu Ala Glu Tyr Phe Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Ser Gly Trp Val Phe Gly Gly Gly Thr Gln Leu Thr Val Leu 100 105 110256124PRTArtificial SequenceSynthetically generated peptide 256Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Gly Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Arg Trp Tyr Asn Asp Tyr Ala 50 55 60Ala Ser Val Lys Ser Arg Ile Thr Val Asn Ala Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Ser Val Arg Tyr Ser Ser Gly Trp Gly Phe Asp 100 105 110Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120257111PRTArtificial SequenceSynthetically generated peptide 257Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Ile Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Phe Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Val Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Ser Gly Arg Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110258124PRTArtificial SequenceSynthetically generated peptide 258Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Arg Asn 20 25 30Ser Ala Ala Trp Asn Leu Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Thr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Arg Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Ser Gly Gly Gly His Ala Ala Gly Lys Phe Asp 100 105 110Ser Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser 115 120259110PRTArtificial SequenceSynthetically generated peptide 259Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Pro Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Lys 20 25 30Tyr Val Tyr Trp Tyr Gln His Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Ser Ala Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110260120PRTArtificial SequenceSynthetically generated peptide 260Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Thr Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Met Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Asp Asp Arg Ala Val 85 90 95Tyr Tyr Cys Arg Gly Arg Leu Gly Gly Gly Met Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Thr Val Ser Ser 115 120261108PRTArtificial SequenceSynthetically generated peptide 261Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Asn Tyr Val 20 25 30Tyr Trp Tyr Gln His Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr 35 40 45Thr Asn Asn Arg Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser 50 55 60Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg Ser Glu65 70 75 80Asp Glu Ala Asp Tyr Phe Cys Ala Ala Trp Asp Asp Ser Leu Ser Gly 85 90 95Trp Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu 100 105262121PRTArtificial SequenceSynthetically generated peptide 262Gln Val Gln Leu Val Gln Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Gly Val Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg Val Thr Thr Gly Ile Thr Arg Tyr Phe Asp Leu Trp Gly 100 105 110Arg Gly Thr Leu Val Thr Val Ser Ser 115 120263111PRTArtificial SequenceSynthetically generated peptide 263Gln Ser Val Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln1 5 10 15Ser Ile Thr Ile Ser Cys Thr Gly Thr Asn Ser Asp Ile Gly Gly Tyr 20 25 30Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45Met Ile Phe Glu Val Thr Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Ala Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80Gln Ala Asp Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser 85 90 95Asn Thr Pro Ser Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu 100 105 110264120PRTArtificial SequenceSynthetically generated peptide 264Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Met 85 90 95Tyr Tyr Cys Val Arg Ser Gly Gly Gly Arg Val Asp Pro Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser 115 120265110PRTArtificial SequenceSynthetically generated peptide 265Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Arg Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Arg Asn His Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Ala Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Ser Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu 100 105 110266119PRTArtificial SequenceSynthetically generated peptide 266Gln Val Gln Leu Val Gln Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asp Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Thr Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asp Ser Leu Arg Pro Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Arg Glu Ala Ser Ser Gly Trp Tyr Ile Asp Ser Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser 115267109PRTArtificial SequenceSynthetically generated peptide 267Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Gln 85 90 95Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Val Lys Ser 100 105268125PRTArtificial SequenceSynthetically generated peptide 268Arg Val Gln Leu Gln Gln Leu Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Phe Gly Asp Ser Val Ser Ser Asn 20 25 30Gly Ala Ala Trp Ser Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ala Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Arg Gly Arg Ile Thr Ile Asn Ala Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Arg Val Val Ser Ser 100 105 110Gly Tyr Trp Gly Gln Gly Thr Leu Val Ala Val Ser Ser 115 120 125269110PRTArtificial SequenceSynthetically generated peptide 269Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Ser Asn Arg Arg Pro Ser Gly Val Pro Gly Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu 100 105 110270125PRTArtificial SequenceSynthetically generated peptide 270Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Pro1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Val Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Gly Tyr Ala 50 55 60Val Ser Val Arg Gly Arg Ile Thr Thr Asn Ala Asp Thr Ser Arg Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Ser 100 105 110Asn Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125271110PRTArtificial SequenceSynthetically generated peptide 271Gln Pro Ala Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Lys Asn Asn Gln Arg Pro Ser Gly Val Pro Gly Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ala Ala Ser Leu Ala Ile Ser Gly Leu Arg65 70 75 80Ser Lys Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu 100 105 110272258PRTArtificial SequenceSynthetically generated peptide 272Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Thr Lys Trp Tyr Asn Glu Tyr Ala 50 55 60Ala Ser Val Lys Gly Arg Ala Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Thr Asp Pro Lys Gly Val Thr Thr Gln Tyr Trp Gly 100 105 110Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala Ser Ala Ser Thr 115 120 125Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 130 135 140Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly145 150 155 160Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly Ser Thr Ser Asn 165 170 175Ile Gly Arg Asn Tyr Val Tyr Trp Tyr Gln Arg Leu Pro Gly Thr Ala 180 185 190Pro Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Ala Pro 195 200 205Ala Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Thr Ser Leu Ala Ile 210 215 220Ser Gly Leu Arg Ser Glu Asp Glu Ala Glu Tyr Phe Cys Ala Ala Trp225 230 235 240Asp Asp Ser Leu Ser Gly Trp Val Phe Gly Gly Gly Thr Gln Leu Thr 245 250 255Val Leu273255PRTArtificial SequenceSynthetically generated peptide 273Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Gly Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Arg Trp Tyr Asn Asp Tyr Ala 50 55 60Ala Ser Val Lys Ser Arg Ile Thr Val Asn Ala Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Ser Val Arg Tyr Ser Ser Gly Trp Gly Phe Asp 100 105 110Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly 115 120 125Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 130 135 140Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln145 150 155 160Arg Ile Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn

165 170 175Tyr Val Tyr Trp Tyr Gln Gln Phe Pro Gly Thr Ala Pro Lys Leu Leu 180 185 190Val Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 195 200 205Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 210 215 220Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu225 230 235 240Ser Gly Arg Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 245 250 255274254PRTArtificial SequenceSynthetically generated peptide 274Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Arg Asn 20 25 30Ser Ala Ala Trp Asn Leu Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Thr Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Arg Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Ser Gly Gly Gly His Ala Ala Gly Lys Phe Asp 100 105 110Ser Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly 115 120 125Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 130 135 140Gln Pro Val Leu Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln145 150 155 160Arg Val Thr Ile Pro Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Lys 165 170 175Tyr Val Tyr Trp Tyr Gln His Leu Pro Gly Thr Ala Pro Lys Leu Leu 180 185 190Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 195 200 205Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 210 215 220Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu225 230 235 240Ser Ala Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 245 250275248PRTArtificial SequenceSynthetically generated peptide 275Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Thr Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Ser Arg Met Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Asp Asp Arg Ala Val 85 90 95Tyr Tyr Cys Arg Gly Arg Leu Gly Gly Gly Met Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Thr Val Ser Ser Gly Ile Leu Gly Ser Gly Gly Gly 115 120 125Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu 130 135 140Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile145 150 155 160Ser Cys Ser Gly Ser Ser Ser Asn Ile Asn Tyr Val Tyr Trp Tyr Gln 165 170 175His Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Thr Asn Asn Arg 180 185 190Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr 195 200 205Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg Ser Glu Asp Glu Ala Asp 210 215 220Tyr Phe Cys Ala Ala Trp Asp Asp Ser Leu Ser Gly Trp Val Phe Gly225 230 235 240Gly Gly Thr Lys Val Thr Val Leu 245276252PRTArtificial SequenceSynthetically generated peptide 276Gln Val Gln Leu Val Gln Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Gly Val Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Arg Val Thr Thr Gly Ile Thr Arg Tyr Phe Asp Leu Trp Gly 100 105 110Arg Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly Ser Gly Gly 115 120 125Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val 130 135 140Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln Ser Ile Thr145 150 155 160Ile Ser Cys Thr Gly Thr Asn Ser Asp Ile Gly Gly Tyr Asn Tyr Val 165 170 175Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile Phe 180 185 190Glu Val Thr Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Ala Ser 195 200 205Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Asp 210 215 220Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser Asn Thr Pro225 230 235 240Ser Val Phe Gly Thr Gly Thr Lys Leu Thr Val Leu 245 250277250PRTArtificial SequenceSynthetically generated peptide 277Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Lys Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Met 85 90 95Tyr Tyr Cys Val Arg Ser Gly Gly Gly Arg Val Asp Pro Trp Gly Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser Gly Ile Leu Gly Ser Gly Gly Gly 115 120 125Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu 130 135 140Thr Gln Ser Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile145 150 155 160Ser Cys Ser Gly Ser Arg Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp 165 170 175Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Arg Asn 180 185 190His Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Ala Ser Lys Ser 195 200 205Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg Ser Glu Asp Glu 210 215 220Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Ser Gly Tyr Val225 230 235 240Phe Gly Thr Gly Thr Lys Leu Thr Val Leu 245 250278255PRTArtificial SequenceSynthetically generated peptide 278Gln Val Gln Leu Val Gln Ser Glu Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asp Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Thr Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asp Ser Leu Arg Pro Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Arg Glu Ala Ser Ser Gly Trp Tyr Ile Asp Ser Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Gly Ile 115 120 125Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 130 135 140Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser145 150 155 160Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser 165 170 175Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 180 185 190Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg 195 200 205Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 210 215 220Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser225 230 235 240Ser Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Val Lys Ser 245 250 255279262PRTArtificial SequenceSynthetically generated peptide 279Arg Val Gln Leu Gln Gln Leu Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Phe Gly Asp Ser Val Ser Ser Asn 20 25 30Gly Ala Ala Trp Ser Trp Ile Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Ala Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala 50 55 60Val Ser Val Arg Gly Arg Ile Thr Ile Asn Ala Asp Thr Ser Lys Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Arg Val Val Ser Ser 100 105 110Gly Tyr Trp Gly Gln Gly Thr Leu Val Ala Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Ile Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Val Leu Thr Gln Ser Pro145 150 155 160Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu 180 185 190Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Ser Asn Arg Arg Pro 195 200 205Ser Gly Val Pro Gly Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala 210 215 220Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr225 230 235 240Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly 245 250 255Thr Lys Leu Thr Val Leu 260280262PRTArtificial SequenceSynthetically generated peptide 280Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Pro1 5 10 15Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn 20 25 30Ser Ala Ala Trp Asn Trp Val Arg Gln Ser Leu Ser Arg Gly Leu Glu 35 40 45Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Gly Tyr Ala 50 55 60Val Ser Val Arg Gly Arg Ile Thr Thr Asn Ala Asp Thr Ser Arg Asn65 70 75 80Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val 85 90 95Tyr Tyr Cys Ala Arg Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Ser 100 105 110Asn Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Ala 115 120 125Ser Ala Pro Thr Gly Ile Leu Gly Ser Gly Gly Gly Gly Ser Gly Gly 130 135 140Gly Gly Ser Gly Gly Gly Gly Ser Gln Pro Ala Leu Thr Gln Ser Pro145 150 155 160Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys Ser Gly 165 170 175Ser Ser Ser Asn Ile Gly Ser Asn Tyr Val Tyr Trp Tyr Gln Gln Leu 180 185 190Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Lys Asn Asn Gln Arg Pro 195 200 205Ser Gly Val Pro Gly Arg Phe Ser Gly Ser Lys Ser Gly Thr Ala Ala 210 215 220Ser Leu Ala Ile Ser Gly Leu Arg Ser Lys Asp Glu Ala Asp Tyr Tyr225 230 235 240Cys Ala Ala Trp Asp Asp Ser Leu Arg Gly Tyr Val Phe Gly Thr Gly 245 250 255Thr Lys Leu Thr Val Leu 26028118PRTArtificial SequenceSynthetically generated peptide 281Arg Thr Tyr Tyr Arg Thr Lys Trp Tyr Asn Glu Tyr Ala Ala Ser Val1 5 10 15Lys Gly2829PRTArtificial SequenceSynthetically generated peptide 282Asp Pro Lys Gly Val Thr Thr Gln Tyr1 528313PRTArtificial SequenceSynthetically generated peptide 283Ser Gly Ser Thr Ser Asn Ile Gly Arg Asn Tyr Val Tyr1 5 1028412PRTArtificial SequenceSynthetically generated peptide 284Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser1 5 1028511PRTArtificial SequenceSynthetically generated peptide 285Ala Ala Trp Asp Asp Ser Leu Ser Gly Trp Val1 5 1028618PRTArtificial SequenceSynthetically generated peptide 286Arg Thr Tyr Tyr Arg Ser Arg Trp Tyr Asn Asp Tyr Ala Ala Ser Val1 5 10 15Lys Ser28712PRTArtificial SequenceSynthetically generated peptide 287Ser Val Arg Tyr Ser Ser Gly Trp Gly Phe Asp Tyr1 5 1028813PRTArtificial SequenceSynthetically generated peptide 288Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Tyr1 5 1028912PRTArtificial SequenceSynthetically generated peptide 289Lys Leu Leu Val Tyr Arg Asn Asn Gln Arg Pro Ser1 5 1029012PRTArtificial SequenceSynthetically generated peptide 290Ala Ala Trp Asp Asp Ser Leu Ser Gly Arg Trp Val1 5 102917PRTArtificial SequenceSynthetically generated peptide 291Arg Asn Ser Ala Ala Trp Asn1 529218PRTArtificial SequenceSynthetically generated peptide 292Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Thr Asp Tyr Ala Val Ser Val1 5 10 15Lys Ser29312PRTArtificial SequenceSynthetically generated peptide 293Ser Gly Gly Gly His Ala Ala Gly Lys Phe Asp Ser1 5 1029413PRTArtificial SequenceSynthetically generated peptide 294Ser Gly Ser Ser Ser Asn Ile Gly Ser Lys Tyr Val Tyr1 5 1029511PRTArtificial SequenceSynthetically generated peptide 295Ala Ala Trp Asp Asp Ser Leu Ser Ala Trp Val1 5 102967PRTArtificial SequenceSynthetically generated peptide 296Ser Asn Ser Ala Thr Trp Asn1 529710PRTArtificial SequenceSynthetically generated peptide 297Gly Gly Arg Leu Gly Gly Gly Met Asp Val1 5 1029811PRTArtificial SequenceSynthetically generated peptide 298Ser Gly Ser Ser Ser Asn Ile Asn Tyr Val Tyr1 5 1029912PRTArtificial SequenceSynthetically generated peptide 299Lys Leu Leu Ile Tyr Thr Asn Asn Arg Arg Pro Ser1 5 1030017PRTArtificial SequenceSynthetically generated peptide 300Val Ile Ser Tyr Asp Gly Ser Lys Lys Tyr Tyr Ala Asp Ser Val Lys1 5 10 15Gly30112PRTArtificial SequenceSynthetically generated peptide 301Arg Val Thr Thr Gly Ile Thr Arg Tyr Phe Asp Leu1 5 1030214PRTArtificial SequenceSynthetically generated peptide 302Thr Gly Thr Asn Ser Asp Ile Gly Gly Tyr Asn Tyr Val Ser1 5 1030312PRTArtificial SequenceSynthetically generated peptide 303Lys Leu Met Ile Phe Glu Val Thr Asn Arg Pro Ser1 5 1030411PRTArtificial SequenceSynthetically generated peptide 304Ser Ser Tyr Ala Gly Ser Asn Thr Pro Ser Val1 5 1030518PRTArtificial SequenceSynthetically generated peptide 305Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Lys Gly3068PRTArtificial SequenceSynthetically generated peptide 306Ser Gly Gly Gly Arg Val Asp Pro1 530713PRTArtificial

SequenceSynthetically generated peptide 307Ser Gly Ser Arg Ser Asn Ile Gly Ser Asn Tyr Val Tyr1 5 1030812PRTArtificial SequenceSynthetically generated peptide 308Lys Leu Leu Ile Tyr Arg Asn His Gln Arg Pro Ser1 5 1030911PRTArtificial SequenceSynthetically generated peptide 309Ala Ala Trp Asp Asp Ser Leu Ser Gly Tyr Val1 5 103105PRTArtificial SequenceSynthetically generated peptide 310Asp Tyr Ala Met His1 531117PRTArtificial SequenceSynthetically generated peptide 311Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Thr Asp Ser Val Lys1 5 10 15Gly31210PRTArtificial SequenceSynthetically generated peptide 312Glu Ala Ser Ser Gly Trp Tyr Ile Asp Ser1 5 103139PRTArtificial SequenceSynthetically generated peptide 313Gln Gln Tyr Gly Ser Ser Gln Val Thr1 53147PRTArtificial SequenceSynthetically generated peptide 314Ser Asn Gly Ala Ala Trp Ser1 531518PRTArtificial SequenceSynthetically generated peptide 315Arg Ala Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala Val Ser Val1 5 10 15Arg Gly31613PRTArtificial SequenceSynthetically generated peptide 316Thr Gly Tyr Ser Ser Ser Arg Val Val Ser Ser Gly Tyr1 5 1031712PRTArtificial SequenceSynthetically generated peptide 317Lys Leu Leu Ile Tyr Lys Ser Asn Arg Arg Pro Ser1 5 1031818PRTArtificial SequenceSynthetically generated peptide 318Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Gly Tyr Ala Val Ser Val1 5 10 15Arg Gly31913PRTArtificial SequenceSynthetically generated peptide 319Thr Gly Tyr Ser Ser Ser Trp Val Val Asn Ser Asn Tyr1 5 10

Claims

1. A composition comprising an isolated human antibody or a fragment or other variant thereof, wherein the antibody, the fragment, or the other variant specifically binds to an aspartyl (asparaginyl) .beta.-hydroxylase (AAH).