RU2826454C2 - Activated polypeptides of interleukin-2 and methods of use thereof - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to production of fused proteins, which are conditionally active versions of interleukin 2 (IL-2) and can be used in medicine for treating tumours. Disclosed is conditionally active IL-2, containing a fused polypeptide, which includes an IL-2 polypeptide, an element for prolonging half-life and an IL-2 blocking fragment connected by linkers.

EFFECT: invention provides obtaining a conditionally active prodrug of IL-2 with an extended half-life, which delivers active IL-2 after cleavage by a protease, in particular, to the desired sites of activity, such as the tumour microenvironment.

10 cl, 23 dwg, 3 tbl, 13 ex

Description

RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No. 62/671,225, filed

May 14, 2018, U.S. Provisional Application No. 62/756,504, filed November 6, 2018, and U.S. Provisional Application No. 62/756,507, filed November 6, 2018. The entire disclosures of the above applications are incorporated herein by reference.

SEQUENCE LIST

This application contains a Sequence Listing, which was filed electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy created on May 14, 2019 is named 105365-0021_SL.txt and is 408,319 bytes in size.

LEVEL OF TECHNOLOGY

[1] The development of mature immunocompetent lymphoid cells from less mimicked precursors, their subsequent antigen-mediated immune responses, and the suppression of these and unwanted autoreactive responses are highly cytokine dependent and regulated by cytokines (including interleukin-2 [IL-2], IL-4, IL-7, IL-9, IL-15, and IL-21) that utilize receptors in the common γ-chain (γc) family (Rochman et al., 2009) and members of the family including IL-12, 18, and 23. IL-2 is essential for Treg cell development in the thymus and heavily regulates several key aspects of mature peripheral Treg and antigen-activated conventional T cells. Because of its marked activity as a T-cell growth factor in vitro, IL-2 has been studied extensively, partly because this activity provides a potential means to directly enhance immunity, such as in AIDS/HIV patients, or as a target for antagonizing adverse responses such as transplant rejection and autoimmune diseases. Although in vitro studies with IL-2 provide a strong rationale for these studies, it is clear that the function of IL-2 in vivo is much more complex, as was first demonstrated in IL-2-deficient mice that exhibited a rapid lethal autoimmune syndrome rather than a lack of immunity (Sadlacket et al., 1993, 1995). Similar observations were made later with individual ablation of the gene encoding IL-2Rα (Il2ra) and IL-2Rβ (Il2rb) (Suzuki et al., 1995; Willerford et al., 1995).

[2] The present invention relates to conditionally active and/or targeted cytokines for use in the treatment of cancer and other diseases dependent on up- or down-regulation of immunity. For example, the antitumor activity of some cytokines is well known and described, and some cytokines are already used for therapeutic purposes in the treatment of humans. Cytokines such as interleukin-2 (IL-2) have demonstrated positive antitumor activity in patients with various types of tumors, such as metastatic renal cell carcinoma, hairy cell leukemia, Kaposi's sarcoma, melanoma, multiple myeloma, etc. Other cytokines such as IFNβ, tumor necrosis factor (TNF) α, TNFβ, IL-1, 4, 6, 12, 15 and CSF have demonstrated some antitumor activity in the case of some types of tumors and, therefore, are the subject of additional studies.

BRIEF DESCRIPTION OF THE ESSENCE OF THE INVENTION

[3] The present invention provides therapeutic proteins, nucleic acids that encode the proteins, and compositions and methods of using such proteins and nucleic acids for treating a disease or disorder such as a proliferative disease, a neoplastic disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, a graft-versus-host disease, etc.

[4] The present invention relates to fusion proteins that are conditionally active variants of IL-2. In one aspect, the full-length polypeptides of the present invention have reduced or minimal IL-2 receptor activating activity even though they comprise a functional cytokine polypeptide. Once activated, such as by cleavage of a linker that connects a blocking moiety, such as a sterically blocking polypeptide, in sequence with an active cytokine, IL-2 or a functional fragment or mutein thereof, can bind its receptor and signal. If desired, the full-length polypeptides can comprise a blocking polypeptide moiety that also provides additional advantageous properties. For example, the full-length polypeptide can have a blocking polypeptide moiety that also prolongs serum half-life and/or targets the full-length polypeptide to a desired site of IL-2 activity. Alternatively, the full-length fusion polypeptides may comprise a serum half-life extending element and/or a targeting domain that are distinct from the blocking polypeptide fragment. Preferably, the fusion protein comprises at least one element capable of extending circulating half-life in vivo. Preferably, the element is enzymatically removed at the desired site in the body (e.g., by protease cleavage in the tumor microenvironment) to restore the pharmacokinetic properties of the targeting molecule (e.g., IL2 or IFNa) to be substantially similar to the naturally occurring targeting molecule. Preferably, the fusion proteins are targeted to a desired cell or tissue. As described herein, targeting is accomplished through the action of a blocking polypeptide fragment that also binds to the desired target, or through a targeting domain. The domain that recognizes the target antigen on the preferred target (e.g., a tumor-specific antigen) can be attached to the cytokine via a cleavable or non-cleavable linker. When attached via a non-cleavable linker, the targeting domain can further promote retention of the cytokine in the tumor and can be considered a retention domain. The targeting domain does not necessarily have to be directly linked to the loading molecule and can be directly linked to another element of the fusion protein. This is especially true when the targeting domain is attached via a cleavable linker.

[5] In one aspect, a fusion polypeptide is provided that comprises an IL-2 polypeptide or a functional fragment or mutein thereof and a blocking moiety, such as a steric blocking domain. The blocking moiety is fused to the IL-2 polypeptide directly or via a linker and can be separated from the cytokine polypeptide by cleavage (e.g., protease-mediated cleavage) of the fusion polypeptide at or near the fusion site or linker or in the blocking moiety. For example, if the cytokine polypeptide is fused to a blocking moiety via a linker that contains a protease cleavage site, the cytokine polypeptide is separated from the blocking moiety and can bind its receptor following protease-mediated cleavage of the linker. The linker is designed to be cleavable at a site of desired cytokine activity, such as in the tumor microenvironment, to avoid off-target cytokine activity and reduce the overall toxicity of cytokine therapy.

[6] The blocking moiety may also function as a serum half-life extending element. In some embodiments, the fusion polypeptide further comprises a separate serum half-life extending element. In some embodiments, the fusion polypeptide further comprises a targeting domain. In various embodiments, the serum half-life extending element is a water-soluble polypeptide, such as optionally branched or multi-chain polyethyleneglycol (PEG), full-length human serum albumin (HSA) or a fragment that retains binding to FcRn, an Fc fragment, or a nanobody that directly binds to FcRn or to human serum albumin.

[7] In addition to the serum half-life extending features, the pharmaceutical compositions described herein preferably comprise at least one or more targeting domains that bind to one or more target antigens or one or more regions on a single target antigen. It is intended herein that the polypeptide construct of the invention is cleaved, for example, in the pathological microenvironment or in the blood of a subject at a protease cleavage site and that the targeting domain(s) will bind to a target antigen on a target cell. The at least one target antigen is involved in and/or associated with a disease process, disorder or pathological condition. Typical antigens include those associated with a proliferative disease, a neoplastic disease, an inflammatory disease, an immunologic disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, graft-versus-host disease, or host-versus-graft disease.

[8] In some embodiments, the target antigen is a cell surface molecule such as a protein, lipid, or polysaccharide. In some embodiments, the target antigen is on a tumor cell, a virally infected cell, a bacterially infected cell, an injured red blood cell, an arterial platelet cell, or a fibrous tissue cell.

[9] In some cases, the target antigens are expressed on the surface of a diseased cell or tissue, such as a tumor or cancer cell. Target antigens for tumors include, but are not limited to, fibroblast activation protein alpha (FAPa), trophoblast glycoprotein (5T4), tumor-associated calcium signal transducer 2 (Trop2), fibronectin EDB (EDB-FN), fibronectin EIIIB domain, CGS-2, EpCAM, EGFR, HER-2, HER-3, c-Met, FOLR1, and CEA. The pharmaceutical compositions described herein also comprise proteins comprising two antigen-binding domains that bind to two different target antigens known to be expressed in the diseased cell or tissue. Examples of antigen-binding domain pairs include, but are not limited to, EGFR/CEA, EpCAM/CEA, and HER-2/HER-3.

[10] In some embodiments, the targeting polypeptides independently comprise an scFv, a VH domain, a VL domain, a non-Ig domain, or a ligand that specifically binds to a target antigen. In some embodiments, the targeting polypeptides specifically bind to a cell surface molecule. In some embodiments, the targeting polypeptides specifically bind to a tumor antigen. In some embodiments, the targeting polypeptides specifically and independently bind to a tumor antigen selected from at least one of EpCAM, EGFR, HER-2, HER-3, cMet, CEA, and FOLR1. In some embodiments, the targeting polypeptides specifically and independently bind to two different antigens, wherein at least one of the antigens is a tumor antigen selected from EpCAM, EGFR, HER-2, HER-3, cMet, CEA, and FOLR1. In some embodiments, the targeting polypeptide serves as a retention domain and is attached to the cytokine via a non-cleavable linker.

[11] As described herein, the cytokine blocking moiety can bind to IL-2 and thereby block activation of the IL-2 cognate receptor.

[12] This description also relates to nucleic acids, such as DNA, RNA, mRNA, which encode the conditionally active proteins described herein, as well as vectors and host cells that contain such nucleic acids.

[13] This description also relates to pharmaceutical compositions that contain a conditionally active protein, a nucleic acid that encodes the conditionally active protein, and vectors and host cells that contain such nucleic acids. Typically, the pharmaceutical composition contains one or more physiologically acceptable carriers and/or excipients.

[14] This disclosure also relates to therapeutic methods that comprise administering to a subject in need thereof an effective amount of a conditionally active protein, a nucleic acid that encodes a conditionally active protein, vectors or host cells that contain such nucleic acid, and pharmaceutical compositions of any of the above. Typically, the subject has or is at risk of developing a proliferative disease, a neoplastic disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, a graft-versus-host disease, or a host-versus-graft disease.

[15] This disclosure also relates to the use of a conditionally active protein, a nucleic acid that encodes a conditionally active protein, vectors or host cells that contain such a nucleic acid, and pharmaceutical compositions of any of the above components for the treatment of a subject in need thereof. Typically, the subject has or is at risk of developing a proliferative disease, a neoplastic disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, a graft-versus-host disease, or a host-versus-graft disease.

[16] This description also relates to the use of a conditionally active protein, a nucleic acid that encodes a conditionally active protein, vectors or host cells that contain such a nucleic acid, for the manufacture of a medicament for the treatment of a disease such as a proliferative disease, a neoplastic disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, a graft-versus-host disease or a host-versus-graft disease.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

[17] Fig. 1a schematically illustrates a protease-activated cytokine or chemokine that contains a blocking moiety. The blocking moiety may optionally function as a serum half-life extending domain. The left side of the arrow in the image shows that the cytokine is linked to the blocking moiety via a protease-cleavable linker, blocking its ability to bind to the receptor. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at the protease cleavage site on the linker, releasing the blocking moiety and allowing the cytokine to bind to the receptor.

[18] Fig. 1b schematically illustrates a protease-activated cytokine or chemokine in which HSA (a blocking moiety) is directly linked to the cytokine or chemokine of interest and a protease cleavage site is located between HSA and the cytokine or chemokine of interest. The left side of the arrow in the image shows that the cytokine is linked to the blocking moiety via a protease cleavable linker, blocking its ability to bind to the receptor. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at the protease cleavage site on the linker, releasing the blocking moiety and allowing the cytokine to bind to the receptor.

[19] Fig. 1c schematically illustrates a protease-activated cytokine or chemokine in which more than one HSA (blocking moiety) is directly linked to the molecule of interest. Optionally, one or more HSAs can be linked to the cytokine or chemokine via a linker, such as a linker that contains a protease cleavage site. The left side of the arrow in the image shows that the cytokine is linked to the blocking moiety via the protease cleavable linker, which blocks its ability to bind to the receptor. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at the protease cleavage site on the linker, releasing the blocking moiety and allowing the cytokine to bind to the receptor. The cytokine now has similar PK properties to the native cytokine (e.g., has a short half-life).

[20] Fig. 1d schematically illustrates a protease-activated cytokine or chemokine comprising more than one cytokine, of the same type or different types, each linked to the binding domain via a protease-cleavable linker. The left side of the arrow in the image shows that the cytokine is linked to a blocking moiety via a protease-cleavable linker, which blocks its ability to bind to the receptor. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at the protease cleavage site on the linker, releasing the blocking moiety and allowing the cytokine to bind to the receptor.

[21] Fig. 2 schematically illustrates a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety, and a serum half-life extending domain linked via at least one protease cleavable linker. The left side of the arrow in the image shows that the cytokine is linked to the blocking moiety via the protease cleavable linkers, which blocks its ability to bind to the receptor. It is also linked to a separate half-life extending element, which extends the serum half-life. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at the protease cleavage site on the linker, releasing the half-life extending element and the blocking moiety and allowing the cytokine to bind to the receptor. The cytokine now has similar PK properties compared to the native cytokine (e.g. short half-life).

[22] Fig. 3 schematically illustrates a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety, and a targeting domain linked via at least one protease-cleavable linker. The left side of the arrow in the image shows that the cytokine is linked to the blocking moiety and targeting domain via the protease-cleavable linker, blocking its ability to bind to the receptor. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at a protease cleavage site on the linker, releasing the targeting domain and blocking moiety and allowing the cytokine to bind to the receptor.

[23] Fig. 4a schematically illustrates a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety, a targeting domain, and a serum half-life extending domain linked via at least one protease-cleavable linker, wherein the cytokine polypeptide and the targeting domain are linked via the protease-cleavable linker. The left side of the arrow in the image shows that the cytokine polypeptide is linked to the targeting domain, blocking moiety, and half-life extending element via the protease-cleavable linker(s), which blocks its ability to bind to the receptor. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at the protease cleavage site on the linker(s), releasing the half-life extending element, targeting domain, and blocking moiety, allowing the cytokine to bind to the receptor. The cytokine now has similar PK properties to the native cytokine (e.g. short half-life).

[24] Fig. 4b schematically illustrates a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a blocking moiety, a targeting domain, and a serum half-life extending domain linked via at least one protease cleavable linker. The left side of the arrow in the image shows that the cytokine is linked to the targeting domain, blocking moiety, and half-life extending element via the protease cleavable linker(s), blocking its ability to bind to its receptor. The right side of the arrow in the image shows that in an inflammatory or tumor environment, protease cleavage occurs at the protease cleavage site on the linker(s), releasing the half-life extending element and blocking moiety and allowing the cytokine to bind to the receptor. The targeting moiety remains bound, trapping the cytokine in the tumor microenvironment. The cytokine now has similar PK properties to the native cytokine (e.g., short half-life).

[25] Fig. 5 schematically illustrates the structure of the variable domain of the immunoglobulin molecule. The variable domains of both the light and heavy chains of immunoglobulin contain hypervariable loops or complementarity determining regions (CDRs). The three CDRs of the V domain (CDR1, CDR2, CDR3) form a cluster at one end of the beta barrel. The CDRs are loops that connect the beta chains of the B-C, C'-C'', and F-G folds of the immunoglobulin chain, with the lower loops, which connect the beta chains of the AB, CC', C''-D, and E-F folds of the immunoglobulin chain, and the upper loop, which connects the D-E folds of the immunoglobulin chain, being non-CDR loops.

[26] Fig. 6 schematically illustrates a protease-activated cytokine or chemokine comprising a cytokine or chemokine polypeptide, a capping moiety that is a serum albumin binding domain (e.g., dAb), and a protease-cleavable linker. In the illustrated example, loops other than the CDRs in the serum albumin binding domain (e.g., sdAb) can form a binding site for the cytokine IL-2. In this example, the binding site for serum albumin can be formed by the CDRs of the serum albumin binding domain.

[27] Figures 7a-7h are a series of graphs illustrating the activity of exemplary IL-2 fusion proteins in the IL-2-dependent cytotoxic T lymphocyte cell line CTLL-2. Each graph shows the results of an IL-2 proliferation assay quantified using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega). Each proliferation assay was performed with (Figures 7b, 7d, 7f, 7h) or without (Figures 7a, 7c, 7e, 7g) HSA. Each fusion protein contains an anti-HSA binding moiety, and both uncleaved and MMP9 protease-cleaved versions of the fusion protein were used in each assay.

[28] Figures 8a-8f are a series of graphs illustrating the activity of exemplary IL-2 fusion proteins in the IL-2-dependent cytotoxic T lymphocyte cell line CTLL-2. Each graph illustrates the results of an IL-2 proliferation assay quantified using the CellTiter-Glo Luminescent Cell Viability Assay (Promega). Both uncleaved and MMP9 protease-cleaved versions of the fusion protein were used in each assay.

[29] Figures 9a-9z are a series of graphs illustrating the activity of exemplary IL-2 fusion proteins in the IL-2-dependent cytotoxic T lymphocyte cell line CTLL-2. Each graph illustrates the results of an IL-2 proliferation assay quantified using the CellTiter-Glo Luminescent Cell Viability Assay (Promega). Both uncleaved and MMP9 protease-cleaved versions of the fusion protein were used in each assay.

[30] Figure 10 illustrates the results of the protein cleavage assay described in Example 2. The ACP16 fusion protein was separated on an SDS-PAGE gel in cleaved and uncleaved forms. As can be seen in the gel, cleavage was complete.

[31] Figures 11a-11c are a series of graphs illustrating the results of the IL-2 HEK-Blue reporter assay performed on IL-2 and recombinant human IL2 (Rec hIL-2) fusion proteins. The assay was performed based on the quantitative assessment of secreted alkaline phosphatase (SEAP) activity using the QUANTI-Blue reagent (InvivoGen).

[32] Figures 12a-12b are two graphs illustrating the analysis of ACP16 (12a) and ACP124 (12b) in the IL-2 assay with the HEKBlue reporter in the presence of HSA. Circles illustrate the activity of the uncleaved polypeptide, squares illustrate the activity of the cleaved polypeptide. Figure 12c is a graph illustrating the results of the CTLL-2 proliferation assay. CTLL2 cells (ATCC) were plated in suspension at a concentration of 500,000 cells/well in culture medium with or without 40 mg/ml human serum albumin (HSA) and stimulated with a dilution series of activatable hIL2 for 72 h at 37°C and 5% _CO2 . The activity of uncleaved and cleaved activatable ACP16 was assayed. Cleaved, activatable hIL2 was prepared by incubation with active MMP9. Cellular activity was assessed using the CellTiter-Glo luminescent cell viability assay (Promega). Circles illustrate intact fusion protein and squares illustrate protease-cleaved fusion protein.

[33] Figures 13a-13c are three graphs illustrating the results of the ACP16, ACP124 assay in the tumor xenograft model. Figure 13a illustrates tumor volume over time in mice treated with 4.4 pg ACP16 (squares), 17 pg ACP16 (triangles), 70 pg ACP16 (inverted triangles), 232 pg ACP16 (solid circles) and 12 pg wild-type IL-2 (dashed line, triangles) and 36 pg wild-type IL-2 (dashed line, diamonds) for comparison. Vehicle alone is shown as large open circles. The data show that tumor volume decreased over time in a dose-dependent manner in mice treated with higher concentrations of ACP16. Figure 13b illustrates the tumor volume over time in mice treated with 17 μg ACP124 (squares), 70 μg ACP124 (triangles), 230 μg ACP124 (inverted triangles) and 700 μg ACP124. Vehicle alone is shown as large open circles. Fig. 13c illustrates the tumor volume over time in mice treated with 17 μg ACP16 (triangles), 70 μg ACP16 (circles), 232 μg ACP16 (solid circles) and 17 μg ACP124 (dashed line, triangles), 70 μg ACP124 (dashed line, diamonds), 230 μg ACP124 (dashed line, diamonds) for comparison. One vehicle is shown as dark inverted triangles. The data show that in mice treated with ACP16, but not ACP124, tumor volume decreased over time in a dose-dependent manner.

[34] Figures 14a-14c show a series of spaghetti plots illustrating the activity of the fusion proteins in the MC38 xenograft mouse model, corresponding to the data illustrated in Figure 13. Each line in the plots represents one mouse.

[35] Figure 15 is a graph illustrating tumor volume over time in a xenograft mouse model, showing tumor growth in control mice (open circles) and AP16-treated mice (squares).

[36] Figures 16a-16d are a series of survival graphs illustrating the survival of mice over time after treatment with the cleavable fusion proteins. Figure 16a shows data for mice treated with vehicle alone (gray line), 17 pg ACP16 (dark line), and 1 pg ACP124 (dashed line). Figure 16b shows data for mice treated with vehicle alone (gray line), 70 pg ACP16 (dark line), and 70 pg ACP124 (dashed line). Figure 16c shows data for mice treated with vehicle alone (gray line), 232 pg ACP16 (dark line), and 230 pg ACP124 (dashed line). Figure 16d shows data for mice treated with vehicle alone (gray line), 232 pg ACP16 (dark line), and 700 pg ACP124 (dashed line).

[37] Figure 17 shows a series of spaghetti plots illustrating the activity of the fusion proteins in the MC38 xenograft mouse model. Groups of mice were given a total of four doses, except for the three highest doses of APC132, which resulted in lethal toxicity at 1 week/two doses. Illustrated are: vehicle alone (top), 17, 55, 70, and 230 μg ACP16 (full top row), 9, 28, 36, and 119 μg ACP132 (full middle row), and 13, 42, 54, and 177 μg ACP21 (full bottom row). Each line in the plots represents an individual animal.

[38] Figs. 18-23 illustrate the properties of TriTac polypeptides, which serve as examples of protease-cleavable fusion proteins.

DETAILED DESCRIPTION OF THE INVENTION

[39] Disclosed herein are methods and compositions for creating and using constructs containing inducible cytokines. Cytokines are potent immune agonists, making them promising therapeutic agents for oncology. However, cytokines have proven to have a very narrow therapeutic window. Cytokines have a short serum half-life and are also considered very potent. Consequently, therapeutic administration of cytokines results in undesirable systemic effects and toxicity. This has been compounded by the need to administer large amounts of cytokines in order to achieve the desired cytokine levels at the intended site of cytokine action (e.g., tumors). Unfortunately, due to the biology of cytokines and the inability to effectively target and control their activity, cytokines have failed to live up to expectations in terms of clinical benefit in the treatment of tumors.

[40] This paper describes fusion proteins that overcome the problems of toxicity and short half-life that have severely limited the clinical use of cytokines in oncology. These fusion proteins contain cytokine polypeptides that have receptor agonist activity. However, in the context of the fusion protein, the cytokine receptor agonist activity is attenuated and the circulating half-life is prolonged. These fusion proteins contain protease cleavage sites that are cleaved by proteases associated with the desired site of cytokine activity (e.g., the tumor) and are typically enriched or selectively present at the site of desired activity. Thus, the fusion proteins are preferentially (or selectively) and efficiently cleaved at the desired site of activity, restricting the cytokine activity essentially to the desired site of activity, such as the tumor microenvironment. Protease cleavage at a desired site of activity, such as the tumor microenvironment, results in the release of a form of the cytokine from the fusion protein that is much more active as an agonist at the cytokine receptor than the fusion protein (typically at least about 100-fold more active than the fusion protein). The form of the cytokine that is released upon fusion protein cleavage typically has a short half-life that is often substantially similar to the half-life of the naturally occurring cytokine, further restricting the cytokine's activity to the tumor microenvironment. Even though the half-life of the fusion protein is increased, toxicity is greatly reduced or eliminated because the circulating fusion protein is attenuated and the active cytokine is targeted to the tumor microenvironment. The fusion proteins described herein enable, for the first time, the administration of an effective therapeutic dose of a cytokine to treat tumors, wherein the cytokine activity is essentially confined to the tumor microenvironment and the undesirable systemic effects and toxicity of the cytokine are greatly reduced or eliminated.

[41] Unless otherwise indicated, all art-specific terms, abbreviations, and other scientific terminology used herein are intended to have the meanings commonly understood by those skilled in the art to which this invention pertains. In some instances, definitions of terms having known meanings are provided herein for the purposes of clarity and/or ease of reference, and the inclusion of such definitions herein is not necessarily to be construed as representing a difference from what is generally known in the art. The methods and procedures described or referenced herein are generally well known and applicable to those skilled in the art using conventional techniques, such as the widely used molecular cloning techniques described in Sambrook et al., Molecular Cloning: A Laboratory Manual ^4th ed. (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Where appropriate, procedures involving the use of commercially available kits and reagents are generally performed in accordance with the manufacturer's specified protocols and conditions unless otherwise specified.

[42] "Cytokine" is a well-known term in the art that refers to any of a class of immunoregulatory proteins (such as interleukins or interferons) that are secreted by cells, especially of the immune system, and that are modulators of the immune system. Cytokine polypeptides that may be used in the fusion proteins described herein include, but are not limited to, transforming growth factors such as TGF-α and TGF-β (e.g., TGF-beta1, TGF-beta2, TGF-beta3); interferons such as interferon-α, interferon-β, interferon-γ, interferon-kappa, and interferon-omega; interleukins such as IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, and IL-25; tumor necrosis factors such as tumor necrosis factor alpha and lymphotoxin; chemokines (e.g., C-X-C motif chemokine 10 (CXCL10), CCL19, CCL20, CCL21) and granulocyte-macrophage colony-stimulating factor (GM-CS), as well as fragments of such polypeptides that activate cognate receptors for the cytokine (i.e., functional fragments of the above molecules). "Chemokine" is a term in the art that refers to any of a family of small cytokines with the ability to induce targeted chemotaxis in nearby responsive cells.

[43] It is well known that chemokines have short half-lives, often lasting only a few minutes or hours. Even forms of cytokines that have altered amino acid sequences designed to prolong serum half-life while maintaining receptor agonist activity also typically have short serum half-lives. As used herein, a "short-half-life cytokine" refers to a cytokine that is characterized by an essentially brief circulation in the serum of a subject, such as a serum half-life of less than 10, less than 15, less than 30, less than 60, less than 90, less than 120, less than 240, or less than 480 minutes. In the context of this document, a short half-life cytokine includes cytokines whose sequences have not been modified to achieve a longer than normal half-life in the body of a subject and polypeptides that have altered amino acid sequences intended to prolong the serum half-life while maintaining receptor agonist activity. Typically, a short half-life cytokine polypeptide, such as an IL-2 polypeptide, has a serum half-life comparable to naturally occurring IL-2, such as within 5-fold, 4-fold, 3-fold or 2-fold of naturally occurring IL-2. This latter case does not imply the addition of heterologous protein domains, such as a true half-life extending element, such as serum albumin.

[44] "Sortases" are transposases that modify proteins by recognizing and cleaving a carboxy-terminal sorting signal incorporated into or attached to the end of a target protein or peptide. Sortase A catalyzes the cleavage of the LPXTG motif (SEQ ID NO: 125) (where X is any standard amino acid) between the Thr and Gly residues in the target protein, transiently attaching the Thr residue to the active site Cys residue in the enzyme to form an enzyme-thioacyl intermediate. To complete transpeptidation and create a peptide-monomer conjugate, a biomolecule with an N-terminal nucleophilic group, typically an oligoglycine motif, attacks the intermediate, displacing sortase A and joining the two molecules.

[45] As used herein, a "steric blocker" refers to a polypeptide or polypeptide moiety that can be covalently linked to a cytokine polypeptide, directly or indirectly via other moieties such as linkers, for example in the form of a chimeric polypeptide (fusion protein), but is not otherwise covalently linked to the cytokine polypeptide. A steric blocker may bind to the cytokine polypeptide non-covalently, for example through electrostatic, hydrophobic, ionic or hydrogen bonding. A steric blocker typically inhibits or blocks the activity of the cytokine moiety due to its proximity to the cytokine moiety and its comparable size. A steric blocker may also block by recruiting a large protein binding partner. An example of this is an antibody that binds to serum albumin; While the antibody itself may or may not be large enough to block activation or binding itself, the involvement of albumin allows for sufficient steric blocking.

[46] In the context of this document and description, a “half-life extending element” is a portion of a chimeric polypeptide that increases serum half-life and improves PK, for example, by altering its size (e.g., to exceed the renal filtration threshold), shape, hydrodynamic radius, charge, or absorption, biodistribution, metabolism, and elimination parameters.

[47] In the context of this document, the terms “activatable,” “activate,” “induce,” and “inducible” refer to the ability of a protein, i.e., a cytokine, that is part of a fusion protein, to bind its receptor and exert activity following cleavage of additional elements from the fusion protein.

[48] In the context of this document, “plasmids” or “viral vectors” are agents that transfer the described nucleic acids into a cell without destroying it and contain a promoter that provides for expression of the nucleic acid molecule and/or polypeptide in the cells into which they have been delivered.

[49] As used herein, the terms "peptide", "polypeptide" or "protein" are used in a broad sense to refer to two or more amino acids linked by a peptide bond. Protein, peptide and polypeptide are also used interchangeably herein to refer to amino acid sequences. It should be understood that the term polypeptide as used herein does not imply a specific size or number of amino acids comprising the molecule, and that a peptide of the invention may contain up to several amino acid residues or more.

[50] In the context of this document, "subject" may be a vertebrate, more particularly a mammal (e.g., human, horse, cat, dog, cow, pig, sheep, goat, mouse, rabbit, rat, and guinea pig), bird, reptile, amphibian, fish, and any other animal. The term does not imply a specific age or sex. Thus, it is understood that adult and neonatal subjects of both male and female sexes are included.

[51] In the context of this document, the terms "patient" or "subject" may be used interchangeably and may refer to a subject with a disease or disorder (e.g., cancer). The term patient or subject includes humans and veterinary subjects.

[52] As used herein, the terms "treating" or "treating" refer to a method of reducing the effect of a disease or condition or a symptom of a disease or condition. Thus, in the described method, treating may refer to at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or a substantially complete reduction in the severity of the established disease or condition or a symptom of the disease or condition. For example, a method of treating a disease is considered a treatment if there is a 10% reduction in one or more symptoms of the disease in a subject compared to a control. Thus, the reduction may be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percentage reduction between 10% and 100% compared to native or control levels. It should be understood that treatment does not necessarily refer to a cure or complete elimination of the disease, pathological condition, or symptoms of the disease or pathological condition.

[53] As used herein, the terms "prevent" and "preventing" a disease or disorder refer to an action, such as administering a chimeric polypeptide or a nucleic acid sequence encoding a chimeric polypeptide, that occurs before or about the same time that a subject begins to exhibit one or more symptoms of the disease or disorder, that inhibits or slows the onset or worsening of one or more symptoms of the disease or disorder.

[54] As used herein, the terms "reduction," "decrease," or "inhibition" include a change of at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more compared to a suitable control level. Such terms may include, but do not necessarily include, complete elimination of a function or property, such as agonist activity.

[55] "Attenuated cytokine receptor agonist" refers to a cytokine receptor agonist that has reduced receptor agonist activity compared to a naturally occurring cytokine receptor agonist. An attenuated cytokine receptor agonist may have at least about 10X, at least about 50X, at least about 100X, at least about 250X, at least about 500X, at least about 1000X, or less agonist activity compared to a naturally occurring receptor agonist. When a fusion protein that comprises a cytokine polypeptide described herein is described as "attenuated" or having "attenuated activity," it is intended that the fusion protein is an attenuated cytokine receptor agonist.

[56] An "intact fusion protein" is a fusion protein in which no domains have been removed, such as by protease cleavage. A domain may be removed by protease cleavage or other enzymatic activity, but when a fusion protein is "intact", this has not yet happened.

[57] As used herein, a "moiety" refers to a portion of a molecule that has a distinct function in that molecule, and that function can be performed by that moiety in the context of another molecule. A moiety may be a chemical compound with a specific function or a portion of a biological molecule with a specific function. For example, a "blocking moiety" in a fusion protein is a portion of the fusion protein that is capable of blocking the activity of some or all of the fusion protein. This may be a protein domain such as serum albumin. The blocking may be accomplished by a steric blocker or a specific blocker. A steric blocker blocks by size and position rather than by specific binding; an example is serum albumin. A specific blocker blocks by specific interactions with the moiety to be blocked. A specific blocker must be matched to a specific cytokine or active domain; a steric blocker may be used regardless of the load provided it is large enough.

[58] In general, the therapeutic use of cytokines is severely limited by their systemic toxicity. For example, TNF was initially studied for its ability to induce hemorrhagic necrosis in some tumors and for its in vitro cytotoxic effect on various tumor lines, but it was subsequently shown to have strong anti-inflammatory activity, which may, in the case of pathological overproduction, pose a danger to the human body. Since systemic toxicity is a fundamental problem for the use of pharmacologically active amounts of cytokines in humans, new derivatives and therapeutic strategies are currently under testing, the aim of which is to reduce the toxic effects of this class of biological effectors while maintaining their therapeutic efficacy.

[59] IL-2 has both stimulatory and regulatory functions in the immune system and, along with other members of the common γ-chain (γc) cytokine family, is fundamental for immune hemostasis. IL-2 mediates its action by binding to IL-2 receptors (IL-2Rs), which are either trimeric receptors composed of the IL-2Rα (CD25), IL-2Rβ (CD122), and IL-2Rγ (γc, CD132) chains, or dimeric βγ IL-2Rs (1, 3). Both IL-2R variants are capable of signaling upon IL-2 binding. However, trimeric αβγ IL-2R has approximately 10- to 100-fold greater affinity for IL-2 than dimeric βγ IL-2R (3), implying that CD25 mediates high-affinity binding of IL-2 to its receptor but is not critical for signaling. Trimeric IL-2R is expressed on activated T cells and CD4+ forkhead-box P3 (FoxP3)+ T regulatory cells (Tregs), which are responsive to IL-2 in vitro and in vivo. In contrast, antigen-experienced CD8+ cells (memory cells), CD8+ cells with high CD44 levels and a memory (MP) phenotype, and natural killer (NK) cells have high levels of dimeric βγ IL-2R, and these cells also respond vigorously to IL-2 in vitro and in vivo.

[60] Expression of high-affinity IL-2R is essential for T cells to mount a response to the low concentrations of IL-2 that are transiently available in vivo. IL-2Rα expression is absent in naïve and memory T cells, but is induced following antigen activation. IL-2Rβ is constitutively expressed by NK, NKT, and CD8+ memory T cells, but is also induced in naïve T cells following antigen activation, but is much less tightly regulated and is constitutively expressed by all lymphoid cells. Following induction of high-affinity IL-2R by antigen, IL-2R signaling upregulates IL-2Rα expression, in part through Stat5-dependent regulation of Il2ra transcription (Kim et al., 2001). This process represents a mechanism for maintaining high-affinity IL-2R expression and continuing IL-2 signaling as long as a source of IL-2 exists.

[61] IL-2Rα captures IL-2 via a large hydrophobic capture surface surrounded by a polar periphery, resulting in a relatively weak interaction (Kd 10-8 M) with fast association-dissociation kinetics of binding. However, the IL-2Rα-IL-2 binary complex induces a very small conformational change in IL-2 that promotes association with IL-2Rβ via the distinct polar interaction between IL-2 and IL-2Rβ. The pseudo-high affinity of the trimeric IL2/α/β complex (i.e., Kd ~300 pM) clearly indicates that the trimeric complex is more stable than either IL2 bound to the α chain alone (Kd=10 nM) or to the β chain alone (Kd=450 nM), as reported by Ciardelli. In either case, the IL2/α/β trimer then recruits the γ chain into a quaternary complex capable of signaling, facilitated by a large composite binding site on the IL2-bound β chain on the γ chain.

[62] In other words, the IL-2Rα-IL-2Rβ-IL-2 ternary complex then recruits γc through a weak interaction with IL-2 and a stronger interaction with IL-2Rβ to form a stable, high-affinity quaternary IL-2R (Kd 10-11 M, which is 10 pM). Formation of the high-affinity IL-2-IL-2R quaternary complex results in signaling through the tyrosine kinases Jak1 and Jak3, which are associated with IL-2Rβ and γc, respectively (Nelson and Willerford, 1998). The IL-2-IL-2R quaternary complex is rapidly internalized, with rapid degradation of IL-2, IL-2Rβ, and γc, but IL-2Rα being recycled back to the cell surface ( et al., 1995; Yu and Malek, 2001). Thus, those functional activities that require long-term IL-2R signaling require a continuous source of IL-2 to recruit IL-2Rα and form additional IL-2-IL-2R signaling complexes.

[63] Regulatory T cells actively suppress immune activation and prevent pathological autoreactivity and subsequent development of autoimmune disease. The development of drugs and methods for selectively activating T cells for the treatment of autoimmune diseases has been the subject of intense research and, prior to the development of the present invention, approaches that selectively deliver interleukins to the site of inflammation have been largely unsuccessful. Regulatory T cells (Tregs) are a class of CD4+CD25+ T cells that suppress the activity of other immune cells. Tregs are fundamental to the homeostasis of the immune system and play a major role in maintaining tolerance to self-antigens and modulating the immune response to foreign antigens. Many autoimmune diseases, including type 1 diabetes (T1D), systemic lupus erythematosus (SLE), and graft-versus-host disease (GVHD), have been shown to involve deficiencies in Treg cell numbers or Treg function.

[64] Consequently, there is great interest in developing therapeutic options to enhance the number and/or function of Treg cells. One investigational approach to treating autoimmune diseases is the transplantation of autologous, ex vivo expanded Treg cells (Tang, Q., et al., 2013, Cold Spring Harb. Perspect. Med., 3:1-15). Although this approach has shown promise in animal models of the disease and in several early human clinical trials, it requires personalized treatment with the patient's own T cells and is invasive and technically challenging. Another approach is treatment with low doses of interleukin-2 (IL-2). Treg cells are characterized by the expression of high constitutive levels of the high-affinity IL-2 receptor, IL2Rαβγ, which consists of the IL2Rα (CD25), IL2Rβ (CD122), and IL2Rγ (CD132) subunits, and Treg cell growth has been shown to be IL-2 dependent (Malek, T.R., et al., 2010, Immunity, 33:153-65).

[65] In turn, immune activation has also been achieved using IL-2, and recombinant IL-2 (Proleukin®) has been approved for the treatment of certain cancers. High doses of IL-2 are used to treat patients with metastatic melanoma and metastatic renal cell carcinoma with long-term effects on overall survival.

[66] Clinical trials of low-dose IL-2 treatment in patients with chronic GVHD (Koreth, J., et al., 2011, N Engl J Med., 365:2055-66) and HCV-associated autoimmune vasculitis (Saadoun, D., et al., 2011, N Engl J Med., 365:2067-77) have demonstrated increased Treg levels and evidence of clinical efficacy. New clinical trials have been initiated to investigate the efficacy of IL-2 in many other autoimmune and inflammatory diseases. The rationale for the use of so-called low-dose IL-2 was to exploit the high affinity IL-2 trimeric IL-2 receptor, which is constitutively expressed on Tregs, leaving the remaining T cells that do not express the high-affinity receptor inactivated. Aldesleukin (marketed as Proleukin® by Prometheus Laboratories, San Diego, CA), the recombinant form of IL-2 used in these studies, is associated with significant toxicity. Aldesleukin is approved for the treatment of metastatic melanoma and metastatic renal cell carcinoma, but its side effects are so severe that its use is recommended only in a hospital setting with access to an intensive care unit (Web address: www.proleukin.com/assets/pdf/proleukin.pdf).

[67] Clinical trials of IL-2 in autoimmune diseases have investigated low doses of IL-2 to target Treg cells, as Treg cells are responsive to lower concentrations of IL-2 than many other immune cell types due to their expression of IL2R-alpha (Klatzmann D, 2015 Nat Rev Immunol. 15:283-94). However, even these lower doses have resulted in safety and tolerability issues, and current treatment options have used daily subcutaneous injections, either continuously or in 5-day interval courses. Therefore, there is a need for a therapy for autoimmune diseases that increases Treg cell number and function, that targets Treg cells more specifically than IL-2, that is safer and more tolerable, and that is administered less frequently.

[68] One approach that has been proposed to improve the therapeutic index of IL-2-based therapies for autoimmune diseases is the use of IL-2 variants that are selective for Treg cells over other immune cells. IL-2 receptors are expressed on a number of different immune cell types, including T cells, NK cells, eosinophils, and monocytes, and this broad expression profile likely accounts for its pleiotropic effects on the immune system and high systemic toxicity. In particular, activated T effector cells express IL2Rαβγ, as do lung epithelial cells. However, T effector cell activation is directly counter to the goal of down-modulating and regulating the immune response, and activation of lung epithelial cells leads to the known dose-limiting side effects of IL-2, including pulmonary edema. Indeed, the major side effect of high-dose IL-2 immunotherapy is capillary leak syndrome (CLS), which results in accumulation of intravascular fluid in organs such as the lungs and liver, with subsequent pulmonary edema and liver cell injury. There is no treatment for CLS other than discontinuing IL-2. Low-dose IL-2 regimens have been studied in patients to avoid CLS, but at the cost of suboptimal therapeutic outcomes.

[69] According to the literature, pulmonary edema is thought to result from the release of proinflammatory cytokines from IL-2-activated NK cells, with clear evidence that pulmonary edema is a result of direct binding of IL-2 to pulmonary endothelial cells that express low to moderate levels of functional IL-2R αβγ. Also, pulmonary edema associated with IL-2 interaction with pulmonary endothelial cells was inhibited by blocking CD25 binding with anti-CD25 monoclonal antibody (mAb) in CD25-deficient host mice or by administering CD122-specific IL-2/anti-IL-2 mAb (IL-2/mAb) complexes, thereby preventing pulmonary edema.

[70] Treatment with cytokines other than IL-2 is more limited. IL-15 exhibits immune cell stimulatory activity similar to IL-2 but without the same inhibitory effects, making it a promising immunotherapeutic candidate. Clinical studies of recombinant human IL-15 for the treatment of metastatic malignant melanoma or renal cell carcinoma have demonstrated significant changes in the distribution, proliferation, and activation of immune cells, suggesting potential antitumor activity (Conlon et al., 2014). IL-15 is currently in clinical trials for the treatment of various forms of cancer. However, IL-15 therapy is known to be associated with adverse and toxic effects, such as worsening of certain types of leukemia, graft-versus-host disease, hypotension, thrombocytopenia, and liver injury. (Mishra A., et al., Cance Cell, 2012, 22(5):645-55; Alpdogan O. et al., Blood, 2005, 105(2):866-73; Conlon KC et al., J Clin Oncol, 2015, 33(1):74-82.)

[71] Direct use of IL-2 as an agonist to bind IL-2R and modulate immune responses therapeutically is problematic due to widely described therapeutic risks, such as its short serum half-life and high toxicity. These risks also limit the therapeutic development and use of other cytokines. New forms of cytokines that mitigate these risks are needed. Disclosed herein are compositions and methods involving the use of IL-2 and IL-15 and other cytokines, functional fragments and muteins of cytokines, and conditionally active cytokines designed to address these risks and provide needed immunomodulatory therapeutics.

[72] The present invention is designed to address the shortcomings of direct IL-2 therapy and therapy using other cytokines, for example, by using cytokine blocking moieties, such as steric blocking polypeptides, serum half-life extending polypeptides, targeting polypeptides, junctional polypeptides containing protease cleavable linkers, and combinations thereof. Cytokines including interleukins (e.g. IL-2, IL-7, IL-12, IL-15, IL-18, IL-21 IL-23), interferons (IFN including IFN-alpha, IFN-beta and IFN-gamma), tumor necrosis factors (e.g. TNF-alpha, lymphotoxin), transforming growth factors (e.g. TGF-beta1, TGF-beta2, TGF-beta3), chemokines (C-X-C motif chemokine 10 (CXCL10), CCL19, CCL20, CCL21) and granulocyte macrophage colony stimulating factor (GM-CS) are very effective when administered to patients. As used herein, "chemokine" refers to a family of small cytokines with the ability to induce targeted chemotaxis in nearby responsive cells. Cytokines can provide powerful therapy, but they are accompanied by adverse events that are difficult to control clinically and which have limited the clinical use of cytokines. The present invention relates to new forms of cytokines that can be used in patients, with reduced or eliminated adverse events. In particular, the present invention relates to pharmaceutical compositions containing chimeric polypeptides (fusion proteins), nucleic acids encoding fusion proteins, and pharmaceutical formulations of the above, which contain cytokines or active fragments or muteins of cytokines that have reduced cytokine receptor activating activity compared to the corresponding cytokine. In this case, under selected conditions or in a selected biological environment, the chimeric polypeptides activate their cognate receptors, often with the same or greater efficiency than the corresponding naturally occurring cytokine. As described herein, this is typically achieved using a cytokine blocking moiety that blocks or inhibits the receptor-activating function of a cytokine, its active fragment or mutein, under general conditions but not under selected conditions such as those present at a required site of cytokine activity (e.g., an inflammatory or tumor site).

[73] Chimeric polypeptides and nucleic acids encoding chimeric polypeptides can be created using any suitable method. For example, nucleic acids encoding a chimeric polypeptide can be created using recombinant DNA technology, synthetic chemistry, or a combination of these methods, and expressed in a suitable expression system, such as CHO cells. Likewise, chimeric polypeptides can be created, for example, by expressing a suitable nucleic acid using synthetic or semi-synthetic chemistry, and the like. In some embodiments, the blocking moiety can be attached to the cytokine polypeptide via sortase-mediated conjugation. "Sortases" are transposases that modify proteins by recognizing and cleaving a carboxy-terminal sorting signal incorporated into or attached to the end of a target protein or peptide. Sortase A catalyzes the cleavage of the LPXTG motif (SEQ ID NO: 125) (where X is any standard amino acid) between the Thr and Gly residues in the target protein, transiently attaching the Thr residue to the active site Cys residue in the enzyme to form an enzyme-thioacyl intermediate. To complete the transpeptidation and create a peptide-monomer conjugate, a biomolecule with an N-terminal nucleophilic group, typically an oligoglycine motif, attacks the intermediate, displacing sortase A and joining the two molecules.

[74] To form a cytokine-blocking fragment fusion protein, a cytokine polypeptide is first labeled at the N-terminus with a polyglycine sequence or, alternatively, at the C-terminus with an LPXTG motif (SEQ ID NO: 125). The blocking fragment or other element contains appropriate peptide attachments that serve as acceptor sites for the labeled polypeptides. For conjugation to domains bearing the LPXTG acceptor peptide (SEQ ID NO: 125) attached at the N-terminus, the polypeptide is labeled with an N-terminal polyglycine region. For conjugation to a domain bearing a polyglycine peptide attached at the C-terminus, the polypeptide is labeled at the C-terminus with the LPXTG sortase recognition sequence (SEQ ID NO: 125). Recognizing the polyglycine sequence and LPXTG (SEQ ID NO: 125), sortase forms a peptide bond between the polymer-peptide and the labeled polypeptides. The reaction involving sortase cleaves the glycine residue as an intermediate and occurs at room temperature.

[75] A number of mechanisms can be used to eliminate or reduce the inhibition caused by a blocking moiety. For example, pharmaceutical compositions can comprise an IL-2 polypeptide and a blocking moiety, such as a steric blocking moiety, wherein a protease cleavable linker comprising a protease cleavage site is located between the IL-2 polypeptide and the IL-2 blocking moiety or within the cytokine blocking moiety. If the protease cleavage site is cleaved, the blocking moiety can dissociate from the cytokine, and the cytokine can then activate the cytokine receptor. The cytokine moiety can also be blocked by a specific blocking moiety, such as an antibody, that binds an epitope located on the corresponding cytokine.

[76] Any suitable linker can be used. For example, the linker can comprise glycine-glycine, a sortase recognition motif, or a sortase recognition motif and the peptide sequence (Gly ₄ Ser) _n (SEQ ID NO: 126) or (Gly ₃ Ser) _n , (SEQ ID NO: 127), wherein n is 1, 2, 3, 4, or 5. Typically, the sortase recognition motif comprises the peptide sequence LPXTG (SEQ ID NO: 125), wherein X is any amino acid. In some embodiments, the covalent linkage is between a reactive lysine residue attached to the C-terminus of the cytokine polypeptide and a reactive aspartic acid residue attached to the N-terminus of the blocker or other domain. In other embodiments, the covalent linkage is between a reactive aspartic acid residue attached to the N-terminus of the cytokine polypeptide and a reactive lysine residue attached to the C-terminus of the blocker or other domain.

[77] Accordingly, as described in detail herein, useful cytokine blocking moieties (e.g., IL-2 blocking moieties) may be steric blockers. As used herein, a "steric blocker" refers to a polypeptide or polypeptide moiety that may be covalently linked to a cytokine polypeptide, directly or indirectly via other moieties such as linkers, such as in the form of a chimeric polypeptide (fusion protein), but is not otherwise covalently linked to the cytokine polypeptide. A steric blocker may bind to the cytokine polypeptide non-covalently, such as through electrostatic, hydrophobic, ionic, or hydrogen bonding. A steric blocker typically inhibits or blocks the activity of a cytokine moiety due to its proximity to the cytokine moiety and its comparable size. Steric inhibition of a cytokine can be eliminated by sterically separating the cytokine moiety from the steric blocker, such as by enzymatically cleaving a fusion protein that contains the steric blocker and the cytokine polypeptide at the site between the steric blocker and the cytokine polypeptide.

[78] As described in more detail herein, the blocking function may be combined with or provided by the presence of additional functional components in the pharmaceutical composition, such as a targeting domain, a serum half-life extending element, and protease-cleavable binding polypeptides. For example, a serum half-life extending polypeptide may also be a steric blocker.

[79] Various elements ensure the delivery and activity of IL-2 preferentially to the site of required IL-2 activity and strict limitation of systemic exposure to the interleukin through a blocking and/or targeting strategy, primarily in conjunction with a serum half-life extension strategy. In this serum half-life extension strategy, the blocked version of the interleukin circulates for an extended time (usually 12 weeks or more), but the activated version has the typical serum half-life of the interleukin.

[80] Compared to the serum half-life extended version, the serum half-life of intravenously administered IL-2 is only ~10 due to distribution throughout the total extracellular space of the body, which is large at ~15 L in the average adult. Therefore, IL-2 is metabolized by the kidneys with a half-life of ~2.5 hours. (Smith, K. "Interleukin 2 immunotherapy." Therapeutic Immunology 240 (2001)). Other measurements show that IL-2 has a very short plasma half-life of 85 minutes for intravenous administration and 3.3 hours for subcutaneous administration (Kirchner, G.I., et al., 1998, Br J Clin Pharmacol. 46:5-10). In some embodiments of this invention, the half-life extending element is linked to the interleukin via a linker that is cleaved at the site of action (e.g., by inflammation-specific or tumor-specific proteases) to release the full activity of the interleukin at the desired site and also separate it from the half-life extending element of the uncleaved version. In such embodiments, the fully active and free interleukin will have very different pharmacokinetic (PK) properties - a half-life of hours instead of weeks. In addition, exposure to the active cytokine is limited to the site of the desired cytokine activity (e.g., the site of inflammation or tumor), and systemic exposure to the active cytokine and associated toxicity and side effects are reduced.

[81] Other cytokines contemplated in this invention have similar pharmacology (e.g., IL-15 according to Blood 2011 117:4787-4795; doi: doi.org/10.1182/blood-2010-10-311456) to IL-2 and, accordingly, the design of this invention addresses the problems of using these agents directly and provides chimeric polypeptides that may have an extended half-life and/or be targeted to the site of desired activity (e.g., an inflammatory site or a tumor).

[82] If desired, IL-2 can be engineered to bind the IL-2R complex as a whole or one of the three IL-2R subunits in particular with an affinity that differs from the affinity of the corresponding wild-type IL-2, such as to selectively activate Tregs or Teff. For example, IL-2 polypeptides that have a greater affinity for the trimeric form of the IL-2 receptor over the dimeric beta/gamma form of the IL-2 receptor compared to wild-type IL-2 can have an amino acid sequence that comprises one of the following groups of mutations relative to SEQ ID NO: 1 (the mature IL-2 protein comprising amino acids 21-153 of human IL-2, having Uniprot Accession No. P60568-1): (a) K64R, V69A, and Q74P; (b) V69A, Q74P, and T101A; (c) V69A, Q74P and I128T; (d) N30D, V69A, Q74P and F103S; (e) K49E, V69A, A73V and K76E; (f) V69A, Q74P, T101A and T133N; (g) N30S, V69A, Q74P and I128A; (h) V69A, Q74P, N88D and S99P; (i) N30S, V69A, Q74P and I128T; (j) K9T, Q11R, K35R, V69A and Q74P; (k) A1T, M46L, K49R, E61D, V69A and H79R; (1) K48E, E68D, N71T, N90H, F103S and I114V; (m) S4P, T10A, Q11R, V69A, Q74P, N88D and T133A; (n) E15K, N30S Y31H, K35R, K48E, V69A, Q74P and I92T; (o) N30S, E68D, V69A, N71A, Q74P, S75P, K76R and N90H; (p) N30S, Y31C, T37A, V69A, A73V, Q74P, H79R and I128T; (q) N26D, N29S, N30S, K54R, E67G, V69A, Q74P and I92T; (r) K8R, Q13R, N26D, N30T, K35R, T37R, V69A, Q74P and I92T; and (s) N29S, Y31H, K35R, T37A, K48E, V69A, N71R, Q74P, N88D and I89V. This approach can also be used to produce muteins or other cytokines, including interleukins (e.g., IL-2, IL-7, IL-12, IL-15, IL-18, IL-23), interferons (IFNs, including IFN-alpha, IFN-beta, and IFN-gamma), tumor necrosis factors (e.g., TNF-alpha, lymphotoxin), transforming growth factors (e.g., TGF-beta1, TGF-beta2, TGF-beta3), and granulocyte macrophage column stimulating factor (GM-CS). For example, muteins can be produced that have the desired binding affinity for a cognate receptor.

[83] As noted above, any of the mutant IL-2 polypeptides described herein may comprise the sequences described; they may also be limited to sequences described by or otherwise identical to SEQ ID NO: 1. In addition, any of the mutant IL-2 polypeptides described herein may optionally comprise a substitution of the cysteine residue at position 125 with another residue (e.g., serine) and/or may optionally comprise a deletion of the alanine residue at position 1 of SEQ ID NO: 1.

[84] Another approach to improving the therapeutic index of IL-2-based therapy is to optimize the pharmacokinetics of the molecule to maximize Treg cell activation. Early studies of IL-2 action demonstrated that a minimum of 5-6 hours of exposure to effective concentrations of IL-2 is required for IL-2 to stimulate human T cell proliferation in vitro (Cantrell, D.A., et. al., 1984, Science, 224: 1312-1316). When administered to human patients, IL-2 has a very short plasma half-life of 85 minutes for intravenous administration and 3.3 hours for subcutaneous administration (Kirchner, G.I., et al., 1998, Br J Clin Pharmacol. 46:5-10). Because of its short half-life, maintaining IL-2 in circulation at or near the level needed to stimulate T cell proliferation for the required time requires high doses that result in peak IL-2 levels substantially exceeding the EC50 for Treg cells, or frequent administration. These high peak IL-2 levels may activate IL2Rβγ receptors and have other unintended or undesirable effects, such as CKD, as noted above. An IL-2 analog or a multifunctional protein with IL-2 fused to an FcRn binding domain, with a longer circulating half-life than IL-2, may deliver the target drug concentration over a period of time at a lower dose than IL-2 and with lower peak levels. Therefore, such an IL-2 analog would require lower doses or less frequent administration than IL-2 to effectively stimulate Treg cells. Less frequent subcutaneous administration of the IL-2 drug would also be better tolerated by patients. A therapeutic with such characteristics would clinically translate into improved pharmacological efficacy, reduced toxicity, and improved patient compliance. Alternatively, IL-2 or IL-2 muteins (herein "IL-2*") could be selectively targeted to the intended site of action (e.g., sites of inflammation). This targeting could be achieved by one of several strategies, including the addition to the administered agent of domains that contain IL-2 blockers (or muteins) that are cleavable, or by targeting domains, or both.

[85] In some embodiments, IL-2* partial agonists can be engineered to bind with greater or lesser affinity depending on the desired target; for example, IL-2* can be engineered to bind with increased affinity to one or more receptor subunits but not to others. These types of partial agonists, unlike full agonists or absolute agonists, provide the ability to alter signaling properties to an amplitude that provides the desired functional properties without reaching the limits of undesirable properties. Given the varying potency of partial agonists, a repertoire of IL-2 variants can be engineered that exhibit even greater degrees of distinct signaling activity, ranging from near complete agonism to complete antagonism.

[86] In some embodiments, IL-2* has an altered affinity for IL-2Rα. In some embodiments, IL-2* has a higher affinity for IL-2Rα than wild-type IL-2. In other embodiments, IL-2* has an altered affinity for IL-2Rβ. In one embodiment, IL-2* has an increased binding affinity for IL-2Rβ, such as the N-terminus of IL-2Rβ, which eliminates the functional requirement for IL-2Rα. In another embodiment, IL-2* is generated that has an increased binding affinity for IL-2Rβ but exhibits reduced binding to IL-2Rγ, and is therefore defective in IL-2Rβγ heterodimerization and signaling.

[87] Blocking moieties, described in further detail below, can also be used to promote binding to or activation of one or more receptors. In one embodiment, blocking moieties are added in a manner that blocks binding to or activation of IL-2Rβγ, but does not alter binding to or activation of IL-2Rα. In another embodiment, blocking moieties are added in a manner that reduces binding to or activation of IL-2Rα. In another embodiment, blocking moieties are added in a manner that inhibits binding to or activation of all three receptors. This blocking can be relieved by removing the blocking moieties in a specific environment, such as by proteolytic cleavage of the linkers linking one or more blocking moieties to the cytokine.

[88] A similar approach could be used to improve other cytokines, particularly for their use as immunostimulatory agents, such as in cancer treatment. For example, in this aspect, the pharmacokinetics and/or pharmacodynamics of a cytokine (e.g. IL-2, IL-7, IL-12, IL-15, IL-18, IL-21 IL-23, IFN-alpha, IFN-beta, IFN-gamma, TNF-alpha, lymphotoxin, TGF-beta1, TGF-beta2, TGF-beta3, GM-CSF, CXCL10, CCL19, CCL20 and CCL21) can be selected to maximize the activation of effector cells (e.g. effects on T cells, NK cells) and/or cytotoxic cells that stimulate the immune response (e.g. induction of dendritic cell maturation) at the site of desired activity, such as a tumor, but preferably not systemically.

[89] Thus, provided herein are pharmaceutical compositions comprising at least one cytokine polypeptide, such as interleukins (e.g., IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, IL-23), interferons (IFNs, including IFN-alpha, IFN-beta and IFN-gamma), tumor necrosis factors (e.g., TNF-alpha, lymphotoxin), transforming growth factors (e.g., TGF-beta1, TGF-beta2, TGF-beta3), chemokines (e.g., CXCL10, CCL19, CCL20, CCL21) and granulocyte macrophage colony stimulating factor (GM-CS) or a functional fragment or mutein of any of the above. The polypeptide typically also comprises at least one linker amino acid sequence, wherein the amino acid sequence, in certain embodiments, is cleavable by an endogenous protease. In one embodiment, the linker comprises the amino acid sequence HSSKLQ (SEQ ID NO: 25), GPLGVRG (SEQ ID NO: 128), IPVSLRSG (SEQ ID NO: 129), VPLSLYSG (SEQ ID NO. 130), or SGESPAYYTA (SEQ ID NO: 131). In other embodiments, the chimeric polypeptide further comprises a blocking moiety, such as a steric blocking polypeptide moiety capable of blocking the activity of the interleukin polypeptide. The blocking moiety, for example, can comprise a human serum albumin (HSA) binding domain or optionally a branched or multi-unit polyethyleneglycol (PEG). In an alternative embodiment, the pharmaceutical composition comprises a first cytokine polypeptide or fragment thereof and a blocking moiety, such as a steric blocking polypeptide moiety, wherein the blocking moiety blocks the activity of the cytokine polypeptide or cytokine receptor, and wherein, in certain embodiments, the blocking moiety comprises a protease cleavable domain. In some embodiments, blocking and reducing the activity of the cytokine is achieved simply by attaching additional domains with very short linkers to the N- or C-terminus of the interleukin domain. In such embodiments, it is contemplated that the blocking is relieved by protease cleavage of the blocking moiety or short linker that binds the blocker to the interleukin. Once the domain is truncated or released, it is no longer capable of blocking the activity of the cytokine.

[90] A pharmaceutical composition, such as a chimeric polypeptide, may comprise two or more cytokines, which may be a polypeptide of the same cytokine or polypeptides of different cytokines. For example, the two or more types of cytokines have complementary functions. In some examples, the first cytokine is IL-2 and the second cytokine is IL-12. In some embodiments, each of the polypeptides of the two or more different types of cytokines has an activity that modulates the activity of the polypeptides of other cytokines. In some examples of chimeric polypeptides that comprise polypeptides of two cytokines, the polypeptide of the first cytokine activates T cells and the polypeptide of the second cytokine does not activate T cells. In some examples of chimeric polypeptides that comprise polypeptides of two cytokines, the first cytokine is a chemoattractant, such as CXCL10, and the second cytokine is an activator of immune cells.

[91] Preferably, the cytokine polypeptides (including functional fragments thereof) that comprise the fusion proteins described herein are not mutated or engineered to alter the properties of the naturally occurring cytokine, including receptor binding affinity and specificity or serum half-life. However, changes in amino acid sequence relative to the naturally occurring cytokine (including wild type) are acceptable, for example, to facilitate cloning and to provide the necessary expression levels.

CD25 binding

[92] Modified IL-2 constructs often have impaired CD25 binding. In contrast, the IL-2 polypeptides described herein are preferably not modified to prevent CD25 binding. Preferably, the IL-2 polypeptides described herein bind CD25. Typically, the IL-2 fusion proteins described herein are capable of binding CD25 and block interactions with IL-2R-beta and gamma (CD 122 and CD 132).

Blocking fragment

[93] The blocking moiety may be any moiety that inhibits the ability of a cytokine to bind and/or activate its receptor. The blocking moiety may inhibit the ability of a cytokine to bind and/or activate its receptor by steric blocking and/or non-covalent binding of the cytokine. Examples of suitable blocking moieties include a full-length or cytokine-binding fragment or a mutein of a cognate receptor of a cytokine. Antibodies and fragments thereof, including a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a single-chain variable fragment (scFv), a single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL), and a camelid nanobody variable domain (VHH), dAb, etc., that bind a cytokine may also be used. Other antigen-binding domains that bind a cytokine that can be used include proteins other than immunoglobulins that mimic the binding and/or structure of an antibody, such as anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, darpins, phynomers, Kunitz domain peptides, monobodies, and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin, and CTLA4 scaffolds. Additional examples of suitable blocking polypeptides include polypeptides that sterically inhibit or block the binding of a cytokine to its cognate receptor. Advantageously, such moieties can also function as half-life extending elements. For example, a peptide modified by conjugation to a water-soluble polymer such as PEG can sterically inhibit or prevent the binding of a cytokine to its receptor. Polypeptides or fragments thereof that have a long serum half-life, such as serum albumin (human serum albumin), immunoglobulin Fc, transferrin, etc., as well as fragments and muteins of such polypeptides, can also be used. Antibodies and antigen-binding domains that bind, for example, to a protein with a long serum half-life, such as HSA, immunoglobulin or transferrin, or to a receptor that returns to the plasma membrane, such as FcRn or the transferrin receptor, can also inhibit the cytokine, in particular when binding to the antigen. Examples of such antigen-binding polypeptides include a single-chain variable fragment (scFv), a single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL), and a camelid nanobody variable domain (VHH), dAb, and the like. Other suitable antigen-binding domains that bind a cytokine and that can also be used include proteins other than immunoglobulins that mimic the binding and/or structure of an antibody, such as anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, darpins, phynomers, Kunitz domain peptides, monobodies, and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin, and CTLA4 scaffolds.

[94] In illustrative examples, if IL-2 is a cytokine in a chimeric polypeptide, the blocking moiety may be a full-length receptor or a fragment or mutein of the IL-2 receptor alpha chain (IL-2Rα) or IL-2 receptor beta (IL-2Rβ) or gamma chain (IL-2Rγ), a single-domain anti-IL-2 antibody (dAb) or scFv, a Fab, an anti-CD25 antibody or fragment thereof, and an anti-HAS dAb or scFv, etc.

Additional aspects of the present invention

1. A fusion protein comprising a cytokine moiety that is operably linked to a binding moiety, wherein the binding moiety comprises a non-CDR loop and a cleavable linker, wherein the binding moiety is capable of masking the binding of a cytokine to its receptor and/or activation of the receptor by the cytokine.

2. The fusion protein of aspect 1, wherein the binding moiety is a natural peptide, a synthetic peptide, an engineered backbone, or an engineered total whey protein.

3. The fusion protein according to aspect 1 or 2, characterized in that the constructed backbone comprises sdAb, scFv, Fab, VHH, fibronectin type III domain, immunoglobulin-like backbone, DARPin, cystine knot peptide, lipocalin, triple-helix backbone, protein G-related albumin-binding module or DNA or RNA aptamer backbone.

4. A fusion protein according to any of aspects 1-2, characterized in that the binding fragment is capable of binding to total serum protein.

5. A fusion protein according to any of aspects 1-3, wherein the non-CDR loop is derived from a variable domain, a constant domain, a C1-type domain, a C2-type domain, an I-domain or any combination thereof.

6. A fusion protein according to any of aspects 1-4, characterized in that the binding fragment additionally comprises complementarity determining regions (CDRs).

7. The fusion protein according to aspect 5, characterized in that the binding fragment is capable of binding to total serum protein.

8. The fusion protein according to aspect 6, characterized in that the total whey protein is a protein that prolongs half-life.

9. The fusion protein according to aspect 6 or 7, characterized in that the total serum protein is albumin, transferrin, factor XIII or fibrinogen.

10. A fusion protein according to any one of aspects 5-8, wherein the CDR loop provides a binding site specific for total serum protein or an immunoglobulin light chain, or any combination thereof.

11. A fusion protein according to any one of aspects 19, wherein the cleavable linker comprises a cleavage site.

12. The fusion protein according to aspect 10, characterized in that the cleavage site is recognized by a protease.

13. The fusion protein according to aspect 11, characterized in that the binding fragment is associated with a cytokine.

14. The fusion protein according to aspect 11, characterized in that the binding fragment is covalently linked to the cytokine.

15. The fusion protein according to aspect 11 or 14, characterized in that the binding moiety is capable of masking the binding of the cytokine to its target through specific intermolecular interactions between the binding moiety and the cytokine.

16. A fusion protein according to any of aspects 11-14, wherein the non-CDR loop provides a binding site specific for binding of the fragment to a cytokine.

17. A fusion protein according to any one of aspects 11-15, wherein after cleavage of the cleavable linker, the binding fragment is separated from the cytokine and the cytokine binds to its target.

18. A fusion protein according to any one of aspects 1-16, characterized in that the cytokine binds to a cytokine receptor.

19. The fusion protein of aspect 17, wherein the cytokine receptor comprises a cytokine receptor type I, an IL receptor type I, an IL receptor type II, a chemokine receptor or a tumor necrosis factor superfamily receptor.

20. A fusion protein according to any one of aspects 1-18, wherein the cleavable linker comprises a cleavage site.

21. The fusion protein according to aspect 20, characterized in that the cleavage site is recognized by a protease.

22. The fusion protein of aspect 21, wherein the protease cleavage site is recognized by a serine protease, a cysteine protease, an aspartate protease, a threonine protease, a glutamic acid protease, a metalloproteinase, a gelatinase, or an aspartic peptide lyase.

23. The fusion protein of aspect 21, wherein the protease cleavage site is recognized by cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, kallikrein, hK1, hK10, hK15, plasmin, collagenase, collagenase type IV, stromelysin, factor Xa, chymotrypsin-like protease, trypsin-like protease, elastase-like protease, subtilisin-like protease, actinidain, bromelain, calpain, caspase, caspase-3, Mir1-CP, papain, HIV-1 protease, HSV protease, CMV protease, chymosin, renin, pepsin, matriptase, legumain, plasmepsin, nepenthesin, metalloexopeptidase, metalloendopeptidase, matrix metalloproteinase (MMP), MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, ADAM10, ADAM17, ADAM12, urokinase-type plasminogen activator (uPA), enterokinase, prostate-specific target (PSA, hK3), interleukin-1β-converting enzyme, thrombin, FAP (FAP-α), dipeptidyl peptidase or dipeptidyl peptidase IV (DPPIV/CD26), transmembrane serine protease type II (TTSP), neutrophil elastase, cathepsin G, proteinase 3, neutrophil serine protease 4, mast cell chymase, mast cell tryptase, dipeptidyl peptidase and dipeptidyl peptidase IV (DPPIV/CD26).

24. A conditionally active binding protein comprising a binding fragment (M) that comprises a non-CDR loop, a cytokine, and a cleavable linker (L), wherein the non-CDR loop is capable of binding to the cytokine and the binding fragment is capable of inhibiting the binding of the cytokine to its receptor and/or inhibiting the activation of the receptor by the cytokine.

25. A conditionally active binding protein according to aspect 24, characterized in that the binding fragment is capable of binding to a protein that prolongs half-life.

26. A conditionally active binding protein according to aspect 24 or 25, wherein the binding moiety is a natural peptide, a synthetic peptide, an engineered backbone or an engineered total whey protein.

27. A conditionally active binding protein according to aspect 26, wherein the engineered backbone comprises sdAb, scFv, Fab, VHH, fibronectin type III domain, immunoglobulin-like backbone, DARPin, cystine knot peptide, lipocalin, triple-helix backbone, protein G-related albumin binding module or DNA or RNA aptamer backbone.

28. A conditionally active binder according to any of aspects 24-27, wherein the non-CDR loop is derived from a variable domain, a constant domain, a C1-type domain, a C2-type domain, an I-domain, or any combination thereof.

29. A conditionally active binding protein according to any of aspects 24-28, characterized in that the binding fragment further comprises complementarity determining regions (CDRs).

30. A conditionally active binding protein according to any one of aspects 24-29, characterized in that the binding fragment comprises a binding site specific for total serum protein.

31. A conditionally active binding protein according to aspect 30, characterized in that the total serum protein is albumin, transferrin, factor XIII or fibrinogen.

32. A conditionally active binding protein according to any one of aspects 29-31, wherein the CDRs provide a binding site specific for total serum protein or an immunoglobulin light chain, or any combination thereof.

33. A conditionally active binding protein according to any one of aspects 29-32, characterized in that the binding moiety is capable of masking the binding of a cytokine to its target through specific intermolecular interactions between the binding moiety and the cytokine.

34. A conditionally active binding protein according to any one of aspects 29-33, wherein the non-CDR loop provides a binding site specific for binding of the binding moiety to a cytokine.

35. A conditionally active binding protein according to any one of aspects 24-34, characterized in that the cytokine binds to a cytokine receptor.

36. A conditionally active binding protein according to aspect 35, wherein the cytokine receptor comprises a cytokine receptor type I, an IL receptor type I, an IL receptor type II, a chemokine receptor or a tumor necrosis factor superfamily receptor.

37. A conditionally active binding protein according to any one of aspects 24-36, characterized in that the cleavable linker comprises a cleavage site.

38. A conditionally active binding protein according to aspect 37, characterized in that the cleavage site is recognized by a protease.

39. A conditionally active binding protein according to aspect 38, characterized in that the protease cleavage site is recognized by a serine protease, a cysteine protease, an aspartate protease, a threonine protease, a glutamic acid protease, a metalloproteinase, a gelatinase or an aspartic peptide lyase.

40. The conditionally active binding protein of aspect 38, wherein the protease cleavage site is recognized by cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, kallikrein, hK1, hK10, hK15, plasmin, collagenase, collagenase type IV, stromelysin, factor Xa, chymotrypsin-like protease, trypsin-like protease, elastase-like protease, subtilisin-like protease, actinidain, bromelain, calpain, caspase, caspase-3, Mir1-CP, papain, HIV-1 protease, HSV protease, CMV protease, chymosin, renin, pepsin, matriptase, legumain, plasmepsin, nepenthesin, metalloexopeptidase, metalloendopeptidase, matrix metalloproteinase (MMP), MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, ADAM10, ADAM17, ADAM12, urokinase-type plasminogen activator (uPA), enterokinase, prostate-specific target (PSA, hK3), interleukin-1β-converting enzyme, thrombin, FAP (FAP-α), dipeptidyl peptidase or dipeptidyl peptidase IV (DPPIV/CD26), transmembrane serine protease type II (TTSP), neutrophil elastase, cathepsin G, proteinase 3, serine neutrophil protease 4, mast cell chymase, mast cell tryptase, dipeptidyl peptidase and dipeptidyl peptidase IV (DPPIV/CD26).

41. The conditionally active binding protein of aspect 24, further comprising a half-life extending domain linked to a binding moiety, wherein the half-life extending domain provides the binding protein with a safety switch, and wherein upon cleavage of the linker, the binding protein is activated by separating the binding moiety and the half-life extending domain from the cytokine and thereby separating the binding protein from the safety switch.

42. A conditionally active binding protein according to aspect 41, characterized in that the cleavage of the linker occurs in the tumor microenvironment.

43. A conditionally active binding protein comprising a binding moiety that binds a cytokine via a non-CDR loop in the binding moiety, wherein the binding moiety is further linked to a half-life extension domain and comprises a cleavable linker, wherein the binding moiety has an extended half-life prior to its activation by cleavage of the linker, and upon activation, the binding moiety and the half-life extension domain are separated from the cytokine, and the binding protein in the activated state does not have an extended half-life.

44. A conditionally active binding protein according to aspect 43, characterized in that the cleavage of the linker occurs in the tumor microenvironment.

Elements for extending half-life in vivo

[95] Preferably, the chimeric polypeptides comprise an element for extending the in vivo half-life. Increasing the in vivo half-life of therapeutic molecules with naturally short half-lives allows for more acceptable and modifiable dosing regimens without sacrificing efficacy. As used herein, a "half-life extending element" is a portion of a chimeric polypeptide that increases the in vivo half-life and improves the PK, such as by altering its size (e.g., to exceed the renal filtration threshold), shape, hydrodynamic radius, charge, or absorption, biodistribution, metabolism, and elimination parameters. A typical way to improve the PK of a polypeptide is to express in the polypeptide chain an element that binds to receptors that are recycled to the plasma membrane of cells rather than degraded in lysosomes, such as the FcRn receptor on endothelial cells and the transferrin receptor. Three types of proteins, such as human IgG, HSA (or fragments thereof) and transferrin, are present in human serum for much longer periods than would be predicted by their size, which is a function of their ability to bind to receptors that are recycled rather than degraded in the lysosome. These proteins or fragments thereof that retain the FcRn binding property are typically linked to other polypeptides to extend their serum half-life. In one embodiment, the half-life extending member is the human serum albumin (HSA) binding domain. HSA (SEQ ID NO: 2) can also be directly linked to the pharmaceutical compositions or linked via a short linker. Fragments of HSA can also be used. HSA and its fragment can function as both a blocking moiety and a half-life extending member. Human IgG and Fc fragments can also perform a similar function.

[96] The serum half-life extending element may also be an antigen-binding polypeptide that binds to a protein with a long serum half-life, such as serum albumin, transferrin, etc. Examples of such polypeptides include antibodies and fragments thereof, including a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a single-chain variable fragment (scFv), a single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL), and a camelid nanobody variable domain (VHH), dAb, etc. Other suitable antigen-binding domains include proteins other than immunoglobulins that mimic the binding and/or structure of an antibody, such as anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, darpins, phynomers, Kunitz domain peptides, monobodies, and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin, and CTLA4 scaffolds. Additional examples of antigen-binding polypeptides include a ligand for a desired receptor, a ligand-binding portion of a receptor, a lectin, and peptides that bind or associate with one or more target antigens.

[97] Some preferred serum half-life extending elements are polypeptides that contain complementarity determining regions (CDRs) and, optionally, non-CDR loops. Advantageously, such serum half-life extending elements may extend the serum half-life of a cytokine and also function as cytokine inhibitors (e.g., through steric blocking, non-covalent interactions, or a combination thereof) and/or targeting domains. In some cases, the serum half-life extending elements are domains derived from an immunoglobulin molecule (Ig molecule) or engineered protein scaffolds that mimic the structure and/or binding activity of an antibody. The Ig may be of any class or subclass (IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, etc.). The polypeptide chain of the Ig molecule is folded into a series of parallel beta strands connected by loops. In the variable region, three loops constitute the "complementarity determining regions" (CDRs), which determine the antigen-binding specificity of the molecule. The IgG molecule contains at least two heavy (H) chains and two light (L) chains linked together by disulfide bonds, or an antigen-binding fragment thereof. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region contains three domains - CH1, CH2 and CH3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region contains one domain - CL. The VH and VL regions can be further divided into regions of hypervariability, termed complementarity determining regions (CDRs), which are hypervariable in sequence and/or participate in antigen recognition, and/or generally form structurally defined loops that are interspersed with more conserved regions, termed framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments of the invention, at least some or all of the amino acid sequences of FR1, FR2, FR3, and FR4 are part of the "non-CDR loop" of the binding fragments described herein. As illustrated in Fig. 5, the variable domain of an immunoglobulin molecule has multiple beta chains that are arranged in two folds. The variable domains of both the light and heavy chains of immunoglobulin contain hypervariable loops or complementarity determining regions (CDRs). The three CDRs of the V domain (CDR1, CDR2, CDR3) form a cluster at one end of the beta barrel. The CDRs are loops that connect the beta chains of the B-C, C'-C'', and F-G folds of the immunoglobulin chain, with the lower loops that connect the beta chains of the AB, CC', C''-D, and E-F folds of the immunoglobulin chain and the upper loop that connects the D-E folds of the immunoglobulin chain being non-CDR loops. In some embodiments of the invention, at least some amino acid residues of the constant domain, CH1, CH2, and CH3, are part of the "non-CDR loop" of the binding fragments described herein. The non-CDR loops, in some embodiments, comprise one or more of the AB, CD, EF, and DE loops of the C1-type domain of an Ig or Ig-like molecule; the AB, CC', EF, FG, BC, and EC' loops of the C2-type domain of an Ig or Ig-like molecule; and the DE, BD, GF, A(A1A2)B, and EF loops of the I(intermediate)-type domain of an Ig or Ig-like molecule.

[98] Within the variable domain, the CDRs are considered to be responsible for antigen recognition and binding, while the FR residues are considered to be the scaffold for the CDRs. However, in certain cases, some of the FR residues play an important role in antigen recognition and binding. The scaffold regions that influence antigen binding are divided into two categories. The first are FR residues that contact the antigen and are thus part of the binding site, and some of these residues are located close to the CDRs in the sequence. The other residues are those that are located far from the CDRs in the sequence, but are in close proximity to them in the 3-D structure of the molecule, such as in a loop in the heavy chain. A serum half-life extension domain (e.g., a domain that contains the CDRs) can contain at least one non-CDR loop. In some embodiments, the non-CDR loop provides a binding site for binding to a cytokine, a total serum protein, or another target antigen.

[99] The serum half-life extending element, in addition to comprising a CDR, additionally or alternatively comprises a non-CDR loop. In some embodiments, the non-CDR loop is modified to create an antigen-binding site specific for a desired target antigen, such as a total serum protein such as albumin, or for a cytokine moiety or other target antigen. It is understood that various methods can be used to modify the non-CDR loop, such as site-directed mutagenesis, random mutagenesis, insertion of at least one acid that is foreign to the amino acid sequence of the non-CDR loop, amino acid substitution. In some examples, an antigen peptide is inserted into the non-CDR loop. In some examples, an antigen peptide replaces the non-CDR loop. The modification to create an antigen-binding site is, in some cases, only one non-CDR loop. In other cases, more than one non-CDR loop is modified. For example, a modification may be present in any of the non-CDR loops illustrated in Fig. 5, i.e., AB, CC', C'' D, EF, and D-E. In some cases, the modification is in loop DE. In other cases, modifications are in all four loops of AB, CC', C''-D, E-F.

[100] In some examples, the serum half-life extending element has dual binding specificity and comprises CDRs that specifically bind common serum proteins, such as serum albumin, and non-CDR loops that specifically bind and block the cytokine domain. In other examples, the serum half-life extending element comprises CDRs that specifically bind a target antigen, such as a cytokine domain or another target antigen, and non-CDR loops that specifically bind a common serum protein, such as serum albumin. Preferably, the serum half-life extending element inhibits the binding of the cytokine domain to a cognate cytokine receptor, such as through steric hindrance, through specific intermolecular interactions, or a combination thereof.

[101] In some embodiments, the serum half-life extending element directly non-covalently binds to the cytokine and inhibits its activity.

[102] In certain examples, the binding moiety binds to a cytokine via one or more of the AB, CC', C" D, and E-F loops and binds to a total serum protein, such as albumin, via one or more of the BC, C'C'', and FG loops. In certain examples, the binding moiety binds to a total serum protein, such as albumin, via an AB, CC', C" D, or EF loop and binds to a cytokine via a BC, C'C'', or FG loop. In certain examples, the binding moiety binds to a total serum protein, such as albumin, via an AB, CC', C" D, and EF loop and binds to a cytokine via a BC, C'C'', and FG loop. In certain examples, the binding moiety binds to a common serum protein, such as albumin, via one or more of loops AB, CC', C'' D, and E-F, and binds to a cytokine via one or more of loops BC, C'C'', and FG.

[103] Binding moieties are any type of polypeptide. For example, in certain cases, the binding moieties are natural peptides, synthetic peptides, or fibronectin backbones, or engineered common serum proteins. Common serum proteins include, for example, albumin, fibrinogen, or globulin. In some embodiments, the binding moieties are engineered backbones. Engineered backbones include, for example, sdAb, scFv, Fab, VHH, fibronectin type III domain, immunoglobulin-like backbone (proposed in Halaby et al., 1999. Prot Eng 12(7):563-571), DARPin, cystine knot peptide, lipocalin, triple helix backbone, protein G-related albumin binding module, or DNA or RNA aptamer backbone.

[104] In some cases, the serum half-life extending element is linked to the cytokine domain via loops other than the CDRs, and the cytokine domain is further linked to a targeting domain as described herein. In some cases, the serum half-life extending element comprises a binding site for a total serum protein. In some embodiments, the CDRs provide a binding site for a total serum protein. In some examples, the total serum protein is globulin, albumin, transferrin, IgG1, IgG2, IgG4, IgG3, IgA monomer, factor XIII, fibrinogen, IgE, or pentameric IgM. In some embodiments, the CDRs form a binding site for an immunoglobulin light chain, such as a free Igκ light chain or a free Igλ light chain.

[105] One exemplary conditionally active protein is shown in Fig. 6. In the illustrated example, non-CDR loops in a serum albumin binding domain (e.g., dAb) can form a site for the cytokine IL-2. In this example, the binding site for serum albumin can be formed by the CDRs of the serum albumin binding domain.

[106] The serum half-life extending element may be any type of binding domain, including but not limited to, domains from a monoclonal antibody, a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody. In some embodiments, the binding moiety is a single-chain variable fragment (scFv), a single-domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL), and a camelid nanobody variable domain (VHH). In other embodiments, the binding moieties are non-Ig binding domains, i.e., antibody mimetics such as anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, darpins, phynomers, Kunitz domain peptides, and monobodies.

[107] In other embodiments, the serum half-life extending element may be a water-soluble polymer or a peptide that is conjugated to a water-soluble polymer such as PEG. As used herein, "PEG", "polyethylene glycol", and "poly(ethylene glycol)" are interchangeable terms and encompass any non-peptide, water-soluble poly(ethylene oxide). The term "PEG" also refers to a polymer that contains a majority, i.e., greater than ₅₀ %, of the _-OCH2CH2- repeating subunits. As for specific forms, PEG may have any number or range of molecular weights and structures or geometries, such as "branched", "linear", "bifurcated", "multifunctional", etc., as described in more detail below. PEG is not limited to a specific structure and may be linear (e.g., end-capped, such as alkoxy PEG or bifunctional PEG), branched or multi-unit (e.g., bifurcated PEG or PEG attached to a polyol core), have a dendritic (or star-shaped) architecture, in each case with or without one or more degradable bonds. Furthermore, the internal structure of the PEG may have an organization corresponding to any number of repeating patterns and may be selected from the group consisting of a homopolymer, an alternating copolymer, a random copolymer, a block copolymer, an alternating tripolymer, a random tripolymer, and a block tripolymer. PEG can be conjugated to polypeptides and peptides by any suitable method. Typically, a reactive derivative of PEG, such as an N-hydroxysuccinimidyl ester of PEG, is reacted with a peptide or polypeptide that contains amino acids with a side chain that contains an amine, sulfhydryl, carboxylic acid, or hydroxyl functional group, such as cysteine, lysine, asparagine, glutamine, threonine, tyrosine, serine, aspartic acid, and glutamic acid.

Targeting and Retention Domains

[108] In certain applications, it may be necessary to minimize the amount of time the construct resides in a desired location in the body. This can be achieved by including in the chimeric polypeptide (fusion protein) one additional domain that would affect its movement in the body. For example, chimeric nucleic acids can encode a domain that directs the polypeptide to a location in the body, such as tumor cells or an inflammatory site; this domain is called a "targeting domain", and/or encode a domain that retains the polypeptide in a specific location in the body, such as tumor cells or an inflammatory site; this domain is called a "retention domain". In some embodiments, the domain can function as both a targeting domain and a retention domain. In some embodiments, the targeting domain and/or the retention domain are specific for a protease-rich environment. In some embodiments, the encoded targeting domain and/or retention domain are specific for regulatory T cells (Treg), such as targeting the CCR4 or CD39 receptors. Other suitable targeting domain and/or retention domain include those that have a cognate ligand that is overexpressed in inflammatory tissue, such as the IL-1 receptor or the IL-6 receptor. In other embodiments, suitable targeting domain and/or retention domain include those that have a cognate ligand that is overexpressed in tumor tissue, such as Epcam, CEA, or mesothelin. In some embodiments, the targeting domain is linked to the interleukin via a linker that is cleaved at the site of action (e.g., by inflammatory or cancer-specific proteases) to release the full activity of the interleukin at the desired site. In some embodiments, the targeting domain and/or retention domain are linked to the interleukin via a linker that is not cleaved at the site of action (e.g., by inflammatory or cancer-specific proteases), resulting in retention of the cytokine at the desired location.

[109] Preferred antigens are in some cases expressed on the surface of a pathological cell or tissue, such as a tumor or cancer cell. Antigens useful for tumor targeting and retention include, but are not limited to, EpCAM, EGFR, HER-2, HER-3, c-Met, FOLR1, and CEA. The pharmaceutical compositions described herein also comprise proteins comprising two targeting domains and/or retention domains that bind to two different target antigens known to be expressed in a pathological cell or tissue. Examples of antigen-binding domain pairs include, but are not limited to, EGFR/CEA, EpCAM/CEA, and HER-2/HER-3.

[110] Suitable targeting domains and/or retention domains include antigen binding domains such as antibodies and fragments thereof, including a polyclonal antibody, a recombinant antibody, a human antibody, a humanized antibody, a single chain variable fragment (scFv), a single domain antibody such as a heavy chain variable domain (VH), a light chain variable domain (VL), and a camelid nanobody variable domain (VHH), dAb, and the like. Other suitable antigen-binding domains include proteins other than immunoglobulins that mimic the binding and/or structure of an antibody, such as anticalins, affilins, affibody molecules, affimers, affitins, alphabodies, avimers, darpins, phynomers, Kunitz domain peptides, monobodies, and binding domains based on other engineered scaffolds such as SpA, GroEL, fibronectin, lipocalin, and CTLA4 scaffolds. Additional examples of antigen-binding polypeptides include a ligand for a desired receptor, a ligand-binding portion of a receptor, a lectin, and peptides that bind or associate with one or more target antigens.

[111] In some embodiments, the targeting domains and/or retention domains specifically bind to a cell surface molecule. In some embodiments, the targeting domains and/or retention domains specifically bind to a tumor antigen. In some embodiments, the targeting polypeptides specifically and independently bind to a tumor antigen selected from at least one of fibroblast activating protein alpha (FAPa), trophoblast glycoprotein (5T4), tumor-associated calcium signal transducer 2 (Trop2), EDB fibronectin (EDB-FN), fibronectin EIIIB domain, CGS-2, EpCAM, EGFR, HER-2, HER-3, cMet, CEA, and FOLR1. In some embodiments, the targeting polypeptides specifically and independently bind to two different antigens, wherein at least one of the antigens is a tumor antigen selected from fibroblast activating protein alpha (FAPa), trophoblast glycoprotein (5T4), tumor-associated calcium signal transducer 2 (Trop2), EDB fibronectin (EDB-FN), EIIIB domain of fibronectin, CGS-2, EpCAM, EGFR, HER-2, HER-3, cMet, CEA, and FOLR1.

[112] The targeting antigen and/or retention antigen may be a tumor antigen expressed on a tumor cell. Tumor antigens are well known in the art and include, for example, EpCAM, EGFR, HER-2, HER-3, c-Met, FOLR1, PSMA, CD38, BCMA, and CEA. 5T4, AFP, B7-H3, Cadherm-6, CAIX, CD117, CD123, CD138, CD166, CD19, CD20, CD205, CD22, CD30, CD33, CD352, CD37, CD44, CD52, CD56, CD70, CD71, CD74, CD79b, DLL3, EphA2, FGFR2, , FGFR3, GPC3, gpA33, FLT-3, gpNMB, HPV-16 E6, HPV-16 E7, ITGA2, ITGA3, SLC39A6, MAGE, mesothelin, Muc1, Muc16, NaPi2b, nectin-4, P-cadherin, NY-ESO-1, PRLR, PSCA, PTK7, ROR1, SLC44A4, SLTRK5, SLTRK6, STEAP1, TIM1, Trop2, WT1.

[113] The targeting antigen and/or retention antigen may be an immune checkpoint protein. Examples of immune checkpoint proteins include, but are not limited to, CD27, CD137, 2B4, TIGIT, CD155, ICOS, HVEM, CD40L, LIGHT, TIM-1, OX40, DNAM-1, PD-L1, PD1, PD-L2, CTLA-4, CDS, CD40, CEACAM1, CD48, CD70, A2AR, CD39, CD73, B7-H3, B7-H4, BTLA, IDO1, IDO2, TDO, KIR, LAG-3, TIM-3, or VISTA.

[114] The targeting antigen and/or retention antigen may be a cell surface molecule such as a protein, lipid, or polysaccharide. In some embodiments, the targeting antigen and/or retention antigen is on a tumor cell, a virally infected cell, a bacterially infected cell, an injured red blood cell, an arterial platelet cell, an inflammatory or fibrous tissue cell. The targeting antigen and/or retention antigen may include an immune response modulator. Examples of immune response modulators include, but are not limited to, granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (G-CSF), interleukin 2 (IL-2), interleukin 3 (IL-3), interleukin 12 (IL-12), interleukin 15 (IL-15), B7-1 (CD80), B7-2 (CD86), GITRL, CD3, or GITR.

[115] The targeting antigen and/or retention antigen may be a cytokine receptor. Examples of cytokine receptors include, but are not limited to, type I cytokine receptors such as GM-CSF receptor, G-CSF receptor, IL type I receptors, Epo receptor, LIF receptor, CNTF receptor, TPO receptor; type II cytokine receptors such as IFN-alpha receptor (IFNAR1, IFNAR2), IFB-beta receptor, IFN-gamma receptor (IFNGR1, IFNGR2), IL type II receptors; chemokine receptors such as CC chemokine receptors, CXC chemokine receptors, CX3C chemokine receptors, CS chemokine receptors; Tumor necrosis factor superfamily receptors such as TNFRSF5/CD40, TNFRSF8/CD30, TNFRSF7/CD27, TNFRSF1A/TNFR1/CD120a, TNFRSF1B/TNFR2/CD120b; TGF-beta receptors such as TGF-beta receptor 1, TGF-beta receptor 2; Ig superfamily receptors such as IL-1 receptors, CSF-1R, PDGFR (PDGFRA, PDGFRB), SCFR.

Linkers

[116] As noted above, the pharmaceutical compositions comprise one or more linker sequences. The linker sequence serves to provide flexibility between the polypeptides such that, for example, a blocking moiety is capable of inhibiting the activity of a cytokine polypeptide. The linker sequence may be located between any or all of the cytokine polypeptide, the serum half-life extending member, and/or the blocking moiety. As described herein, at least one of the linkers is cleavable by a protease and comprises (one or more) cleavage sites for (one or more) desired proteases. Preferably, the desired protease is enriched in or selectively present at a desired site of cytokine activity (e.g., a tumor microenvironment). Thus, the fusion protein is preferentially or selectively cleaved at a site of desired cytokine activity.

[117] Suitable linkers may be of any length, such as from 1 amino acid (e.g., Gly) to 20 amino acids, from 2 amino acids to 15 amino acids, from 3 amino acids to 12 amino acids, including from 4 amino acids to 10 amino acids, from 4 amino acids to 9 amino acids, from 6 amino acids to 8 amino acids, or from 7 amino acids to 8 amino acids, and may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 amino acids.

[118] The orientation of the components of the pharmaceutical composition largely depends on the design choice and it is understood that many orientation options are possible and it is assumed that they are all covered by this invention. For example, the blocking moiety may be located at the C-terminus or N-terminus of the cytokine polypeptide.

[119] Proteases known to be associated with pathological cells or tissues include, but are not limited to, serine proteases, cysteine proteases, aspartate proteases, threonine proteases, glutamic acid proteases, metalloproteinases, aspartic peptide lyases, serum proteases, cathepsins, cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L, kallikreins, hK1, hK10, hK15, plasmin, collagenase, collagenase type IV, stromelysin, factor Xa, chymotrypsin-like protease, trypsin-like protease, elastase-like protease, subtilisin-like protease, actinidain, bromelain, calpain, caspase, caspase-3, Mir1-CP, papain, HIV-1 protease, HSV protease, CMV protease, chymosin, renin, pepsin, matriptase, legumain, plasmepsin, nepenthesin, metalloexopeptidases, metalloendopeptidases, matrix metalloproteinases (MMPs), MMP1, MMP2, MMP3, MMP8, MMP9, MMP13, MMP11, MMP14, urokinase-type plasminogen activator (uPA), enterokinase, prostate-specific antigen (PSA, hK3), interleukin-1β-converting enzyme, thrombin, FAP (FAP-α), dipeptidyl peptidase, meprins, granzymes and dipeptidyl peptidase IV (DPPIV/CD26). Proteases capable of cleaving amino acid sequences encoded by the chimeric nucleic acid sequences provided herein may, for example, be selected from the group consisting of prostate-specific antigen (PSA), matrix metalloproteinase (MMP), disintegrin and metalloproteinase (ADAM), plasminogen activator, cathepsin, caspase, tumor cell surface protease and elastase. The MMP may, for example, be matrix metalloproteinase 2 (MMP2) or matrix metalloproteinase 9 (MMP9).

[120] Proteases useful in the methods described herein are presented in Table 1, and exemplary proteases and their cleavage sites are presented in Table 1a:

[121] Provided herein are pharmaceutical compositions comprising polypeptide sequences. As with all peptides, polypeptides and proteins, including fragments thereof, it is understood that there may be additional modifications to the amino acid sequence of the chimeric polypeptides (amino acid sequence variants) that do not alter the nature or function of the peptides, polypeptides or proteins. Such modifications include conservative amino acid substitutions and are discussed in more detail below.

[122] The compositions provided herein have the desired function. The compositions comprise at least an IL-2 polypeptide, a blocking moiety, such as a steric blocking polypeptide, and an optional element for extending serum half-life and an optional targeting polypeptide, wherein each polypeptide in the composition is connected by one or more linkers. A first polypeptide, such as an IL-2 mutein, is provided as an active agent. The blocking moiety is provided for the purpose of blocking the activity of an interleukin. The linker polypeptide, such as a protease-cleavable polypeptide, is provided for cleavage by a protease that is specifically expressed in the intended target of the active agent. Optionally, the blocking moiety blocks the activity of the first polypeptide by binding the interleukin polypeptide. In some embodiments, a blocking moiety, such as a steric blocking peptide, is linked to the interleukin via a protease cleavable linker that is cleaved at the site of action (e.g., by inflammatory or tumor-specific proteases) to release full cytokine activity at the desired site.

[123] The protease cleavage site may be a naturally occurring protease cleavage site or an artificially engineered protease cleavage site. An artificially engineered protease cleavage site may be cleavable by more than one protease specific for the desired environment in which the cleavage will occur, such as a tumor. The protease cleavage site may be cleavable by at least one protease, at least two proteases, at least three proteases, or at least four proteases.

[124] In some embodiments, the linker comprises glycine-glycine, a sortase recognition motif, or a sortase recognition motif and the peptide sequence (Gly ₄ Ser) _n (SEQ ID NO: 126) or (Gly ₃ Ser) _n , (SEQ ID NO: 127), wherein n is 1, 2, 3, 4, or 5. In one embodiment, the sortase recognition motif comprises the peptide sequence LPXTG (SEQ ID NO: 125), wherein X is any amino acid. In one embodiment, the covalent linkage is between a reactive lysine residue attached to the C-terminus of the cytokine polypeptide and a reactive aspartic acid residue attached to the N-terminus of the blocking or other moiety. In one embodiment, the covalent linkage is between a reactive aspartic acid residue attached to the N-terminus of the cytokine polypeptide and a reactive lysine residue attached to the C-terminus of the blocking or other moiety.

Splitting and inducibility

[125] As described herein, the activity of a cytokine polypeptide in the context of a fusion protein is attenuated, and protease cleavage at a desired site of activity, such as a tumor microenvironment, results in the release of a form of the cytokine from the fusion protein that is much more active as a cytokine receptor agonist than the fusion protein. For example, the cytokine receptor activating (agonist) activity of a fusion polypeptide can be at least about 10-fold, at least about 50-fold, at least about 100-fold, at least about 250-fold, at least about 500-fold, or at least about 1000-fold less than the cytokine receptor activating activity of the cytokine polypeptide as a single molecule. A cytokine polypeptide that is part of a fusion protein exists as a single molecule when it contains amino acids that are substantially identical to the cytokine polypeptide and contains substantially no additional amino acids and is not associated (covalently or non-covalently) with other molecules. If desired, the cytokine polypeptide as a single molecule may contain certain additional amino acid sequences, such as tags or short sequences that facilitate expression and/or purification.

[126] In other examples, the cytokine receptor activating (agonistic) activity of the fusion polypeptide is at least about 10-fold, at least about 50-fold, at least about 100-fold, at least about 250-fold, at least about 500-fold, or about 1000-fold less than the cytokine receptor activating activity of a polypeptide that comprises a cytokine polypeptide formed by cleavage of a protease-cleavable linker in the fusion protein. In other words, the cytokine receptor activating (agonistic) activity of a polypeptide that comprises a cytokine polypeptide formed by cleavage of a protease-cleavable linker in a fusion protein is at least about 10 times, at least about 50 times, at least about 100 times, at least about 250 times, at least about 500 times, or at least about 1000 times greater than the cytokine receptor activating activity of the fusion protein.

Polypeptide substitutions

[127] The polypeptides described herein may contain components (e.g., a cytokine, a blocking fragment) that have the same amino acid sequence as the corresponding naturally occurring protein (e.g., IL-2, IL-15, HSA), or may have an amino acid sequence that differs from the naturally occurring protein, so long as the necessary function is retained. It is understood that one way to define any modifications and derivatives of the described proteins and nucleic acids encoding them, known or those that may arise, is to define sequence variants in terms of identity to specific known reference sequences. In particular, polypeptides and nucleic acids are described that have at least 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 percent identity to the chimeric polypeptides provided herein. For example, polypeptides or nucleic acids are provided that have at least 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 percent identity to the sequence of any of the nucleic acids or any of the polypeptides described herein. Those skilled in the art know how to determine the identity of two polypeptides or two nucleic acids. For example, the identity can be calculated after aligning the two sequences so that the identity is at the highest level.

[128] Another method of calculating identity can be carried out using algorithms published in the literature. Optimal alignment of sequences for comparison can be carried out using the local identity algorithm of Smith and Waterman (Smith and Waterman Adv. Appl. Math. 2:482 (1981)), the alignment algorithm of Needleman and Wunsch (Needleman and Wunsch, J. Mol. Biol. 48:443 (1970)), the similarity search method of Pearson and Lipman (Proc. Natl. Acad. Sci. USA 85:2444 (1988)), computerized versions of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics software package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual assessment.

[129] The same types of identities can be obtained for nucleic acids, for example, using the algorithms described in Zuker, Science 244:48-52 (1989); Jaeger et al., Proc. Natl. Acad. Sci. USA 86:7706-7710 (1989); Jaeger et al., Methods Enzymol. 183:281-306 (1989), which are incorporated by reference with respect to at least the material relating to nucleic acid alignment. It is understood that any of the methods can generally be used and that in certain cases the results of these different methods may differ, but one skilled in the art will appreciate that if an identity is found by at least one of these methods, the sequences will be considered to have the stated identities and are described herein.

[130] Protein modifications include modifications of the amino acid sequence. Modifications in the amino acid sequence may occur naturally as allelic variations (e.g., due to genetic polymorphism), may arise due to environmental influences (e.g., due to exposure to ultraviolet light), or may be created by human intervention (e.g., by mutagenesis of cloned DNA sequences), such as induced point, deletion, insertion, and substitution mutants. These modifications may result in changes in the amino acid sequence, provide silent mutations, modify a restriction site, or provide other specific mutations. Amino acid sequence modifications generally fall into one or more of three classes: substitution, insertion, or deletion modifications. Insertions include amino- and/or carboxy-terminal fusions, as well as intrasequence insertions of one or more amino acid residues. Insertions are typically smaller than amino- or carboxyl-terminal fusions, e.g., on the order of one to four residues. Deletions are characterized by the removal of one or more amino acid residues from the protein sequence. Typically, no more than about 2-6 residues are deleted at any one site in the protein molecule. Amino acid substitutions are typically single residues but may be at a number of different locations; insertions are typically on the order of 1-10 amino acid residues; and deletions range from 1-30 residues. Deletions and insertions are preferably made in adjacent pairs, i.e., a deletion of 2 residues or an insertion of 2 residues. Substitutions, deletions, insertions, or any combination thereof can be combined to produce the final construct. Mutations should not displace the sequence out of frame and preferably do not create complementary regions that could lead to secondary structure in the mRNA. Substitutional modifications are those in which at least one residue has been removed and another residue has been inserted in its place. Such substitutions are generally made according to Table 2 below and are considered conservative substitutions.

[131] Modifications, including specific amino acid substitutions, are carried out by known methods. For example, modifications can be carried out by site-specific mutagenesis of nucleotides in DNA encoding the polypeptide, thereby obtaining DNA encoding the modification, and then expressing the DNA in a recombinant cell culture. Methods for creating substitution mutations at specified sites in DNA having a known sequence are well known, for example, mutagenesis with M13 primers and PCR mutagenesis.

[132] Modifications can be selected to optimize binding. For example, affinity maturation methods can be used to alter scFv binding by introducing random mutations into the complementarity determining regions (CDRs). Such random mutations can be introduced by a number of methods, including irradiation, chemical mutagens, or error-producing PCR. Multiple rounds of mutation and selection can be performed using, for example, phage display.

[133] The present invention also relates to nucleic acids that encode the chimeric polypeptides described herein and to the use of such nucleic acids to produce the chimeric polypeptides for therapeutic purposes. For example, the present invention includes DNA and RNA molecules (e.g., mRNA, self-replicating RNA) that encode a chimeric polypeptide and to the therapeutic use of such DNA and RNA molecules.

Typical compositions

[134] In exemplary fusion proteins of the present invention, the above-described elements are combined in a number of variations. The variations described in this section are intended as examples and are not intended to be limiting.

[135] In some embodiments, the fusion protein comprises an IL-2 polypeptide, a blocking moiety, and a half-life extending element. In some embodiments, the IL-2 polypeptide is located between the half-life extending element and the blocking moiety. In some embodiments, the IL-2 polypeptide is located N-terminal to the blocking moiety and the half-life extending element. In some such embodiments, the IL-2 polypeptide is located proximal to the blocking moiety; in some such embodiments, the cytokine is located proximal to the half-life extending element. At least one protease cleavable linker must be included in all embodiments so that the IL-2 polypeptide can become active upon cleavage. In some embodiments, the IL-2 polypeptide is located C-terminal to the blocking moiety and the half-life extending element. The additional elements can be attached to one another via a cleavable linker, a non-cleavable linker, or by direct fusion. In some cases, it may be beneficial to include two identical cytokines to facilitate dimerization.

[136] In some embodiments, the blocking domains used are capable of prolonging half-life, and the IL-2 polypeptide is located between two such blocking domains. In some embodiments, the IL-2 polypeptide is located between two blocking domains, one of which is capable of prolonging half-life.

[137] In some embodiments, two cytokines are included in a single construct, at least one of which is IL-2. In some embodiments, each of the cytokines is coupled to two blocking domains (a total of three in one molecule), with the blocking domain between the two cytokines. In some embodiments, one or more additional domains may be included to extend half-life to optimize pharmacokinetic properties.

[138] In some embodiments, three cytokines are included in a single construct. In some embodiments, the third cytokine may have a blocking function for the other two instead of a blocking domain between the two cytokines.

[139] Preferred half-life extending elements for use in fusion proteins include human serum albumin (HSA), an antibody or antibody fragment (e.g., scFV, dAb) that binds serum albumin, human or humanized IgG, or a fragment of any of the above. In some preferred embodiments, the blocking moiety is human serum albumin (HSA) or an antibody or antibody fragment that binds serum albumin, an antibody that binds a cytokine and prevents activation of the binding or activation of the cytokine receptor, another cytokine, or a fragment of any of the above. In preferred embodiments comprising the use of an additional targeting domain, the targeting domain is an antibody that binds to a cell surface protein that is enriched on the surface of cancer cells, such as EpCAM, FOLR1, and fibronectin.

Methods of treatment and pharmaceutical compositions

[140] Further provided are methods of treating a subject that has or is at risk of developing a disease or disorder, such as a proliferative disease, a neoplastic disease, an inflammatory disease, an immunological disorder, an autoimmune disease, an infectious disease, a viral disease, an allergic reaction, a parasitic reaction, or a graft-versus-host disease. The methods include administering to a subject in need thereof an effective amount of a fusion protein described herein, which is typically administered as a pharmaceutical composition. In some embodiments, the method further comprises selecting a subject that has or is at risk of developing such a disease or disorder. The pharmaceutical composition preferably comprises a blocked cytokine, fragment, or mutein thereof that is activated at the site of inflammation or in a tumor. In one embodiment, the chimeric polypeptide comprises a cytokine polypeptide, fragment, or mutein thereof, and an element for extending serum half-life. In another embodiment, the chimeric polypeptide comprises a cytokine polypeptide, a fragment or mutein thereof, and a blocking moiety, such as a steric blocking polypeptide, wherein the steric blocking polypeptide is capable of sterically blocking the activity of the cytokine polypeptide, a fragment or mutein thereof. In another embodiment, the chimeric polypeptide comprises a cytokine polypeptide, a fragment or mutein thereof, a blocking moiety, and an element for extending serum half-life.

[141] Inflammation is part of the complex biological response of body tissues to harmful stimuli such as pathogens, damaged cells, or irritants, and is a protective response involving immune cells, blood vessels, and molecular mediators. The function of inflammation is to eliminate the original cause of cellular injury, clear necrotic cells and tissue damaged by the original insult and the inflammatory process, and initiate tissue repair. Inflammation can occur due to infection, as a symptom of a disease such as cancer, atherosclerosis, allergies, myopathies, HIV, obesity, or an autoimmune disease. An autoimmune disease is a chronic pathological condition that occurs due to an abnormal immune response to self-antigens. Autoimmune diseases that can be treated with the polypeptides described herein include, but are not limited to, lupus, celiac disease, type 1 diabetes mellitus, Graves' disease, inflammatory bowel disease, multiple sclerosis, psoriasis, rheumatoid arthritis, and systemic lupus erythematosus.

[142] The pharmaceutical composition may comprise one or more protease cleavable sequences. The linker sequence serves to provide flexibility between the polypeptides such that each polypeptide is capable of inhibiting the activity of the first polypeptide. The linker sequence may be located between any or all of the cytokine polypeptide, a fragment or mutein thereof, a blocking moiety, and an element for extending serum half-life. Optionally, the composition comprises two, three, four, or five linker sequences. The linker sequence, two, three, or four linker sequences may be the same or different linker sequences. In one embodiment, the linker sequence comprises GGGGS (SEQ ID NO: 132), GSGSGS (SEQ ID NO: 133), or G(SGGG) _2SGGT (SEQ ID NO: 134). In another embodiment, the linker comprises a protease cleavable sequence selected from the group consisting of HSSKLQ (SEQ ID NO: 25), GPLGVRG (SEQ ID NO: 128), IPVSLRSG (SEQ ID NO: 129), VPLSLYSG (SEQ ID NO: 130), and SGESPAYYTA (SEQ ID NO: 131).

[143] In some embodiments, the linker is cleaved by a protease selected from the group consisting of kallikrein, thrombin, chymase, carboxypeptidase A, cathepsin G, elastase, PR-3, granzyme M, calpain, matrix metalloproteinase (MMP), plasminogen activator, cathepsin, caspase, tryptase, or tumor cell surface protease.

[144] Suitable linkers may be of any length, such as from 1 amino acid (e.g., Gly) to 20 amino acids, from 2 amino acids to 15 amino acids, from 3 amino acids to 12 amino acids, including from 4 amino acids to 10 amino acids, from 4 amino acids to 9 amino acids, from 6 amino acids to 8 amino acids, or from 7 amino acids to 8 amino acids, and may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 amino acids.

[145] Further provided are methods for treating a subject that has or is at risk of developing cancer. These methods comprise administering to a subject in need thereof an effective amount of a chimeric polypeptide (fusion protein) described herein, which is typically administered as a pharmaceutical composition. In some embodiments, the method further comprises selecting a subject that has or is at risk of developing cancer. The pharmaceutical composition preferably comprises a blocked cytokine, fragment or mutein thereof that is activated at the site of a tumor. Preferably, the tumor is a solid tumor. The cancer may be, but is not limited to, colon cancer, lung cancer, melanoma, sarcoma, renal cell carcinoma and breast cancer.

[146] The method may further comprise administering one or more additional cancer treatment agents such as chemotherapeutic agents (e.g., adriamycin, cerubidine, bleomycin, alkeran, velban, oncovin, fluorouracil, thiotepa, methotrexate, bisantrene, noanthrone, thioguanine, cytarabine, procarbazine), immuno-oncology agents (e.g., anti-PD-L1, anti-CT1A4, anti-PD-1, anti-CD47, anti-GD2), cell therapy (e.g., CAR-T, T-cell therapy), oncolytic viruses, etc.

[147] Provided herein are pharmaceutical formulations or compositions comprising chimeric polypeptides and a pharmaceutically acceptable carrier. The compositions provided herein are suitable for administration in vitro or in vivo. By pharmaceutically acceptable carrier is meant a material that is not biologically or otherwise undesirable, i.e., the material administered to a subject does not cause undesirable biological effects or does not interact in a harmful manner with other components of the pharmaceutical formulation or composition in which it is contained. The carrier is selected so as to minimize degradation of the active ingredient and minimize undesirable side effects in the subject.

[148] Suitable carriers and formulations thereof are described in Remington: The Science and Practice of Pharmacy, ^21st Edition, David B. Troy, ed., Lippicott Williams & Wilkins (2005). Typically, an appropriate amount of a pharmaceutically acceptable salt is used in the formulation to make the formulation isotonic, although the formulation may be hypertonic or hypotonic as desired. Examples of pharmaceutically acceptable carriers include, but are not limited to, sterile water, saline, buffered solutions such as Ringer's solution and dextrose solution. The pH of the solution is generally from about 5 to about 8 or from about 7 to 7.5. Other carriers include sustained release preparations such as semipermeable matrices of solid hydrophobic polymers containing immunogenic polypeptides. The matrices are in the form of shaped articles such as films, liposomes or microparticles. Certain carriers may be more preferable depending on, for example, the route of administration and the concentration of the composition administered. The carriers are suitable for administering chimeric polypeptides or nucleic acid sequences encoding chimeric polypeptides to humans or other subjects.

[149] The pharmaceutical formulations or compositions are administered by a variety of routes depending on whether local or systemic treatment is desired and the site to be treated. The compositions are administered by any of several routes of administration including topical, oral, parenteral, intravenous, intra-articular, intraperitoneal, intramuscular, subcutaneous, intracavitary, transdermal, intrahepatic, intracranial, by nebulization/inhalation, or by bronchoscopy. In some embodiments, the compositions are administered locally (non-systemically), including intratumorally, intra-articularly, intrathecally, etc.

[150] Parenteral preparations include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous vehicles are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic-aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral carriers include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, Ringer's lactate or fixed oils. Intravenous carriers include fluid and nutrient replenishers, electrolyte replenishers (e.g., Ringer's dextrose), and the like. Preservatives and other additives such as, for example, antimicrobials, antioxidants, chelating agents and inert gases, and the like are optionally present.

[151] Formulations for topical administration include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Traditional pharmaceutical carriers, aqueous, powder or oily bases, thickening agents, etc., are optional or necessary.

[152] Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets or tablets. Optionally, thickening agents, flavoring agents, diluents, emulsifiers, dispersing agents or binders are needed.

[153] Optionally, the chimeric polypeptides or nucleic acid sequences encoding the chimeric polypeptides are administered using a vector. There are a number of compositions and methods that can be used to deliver nucleic acid molecules and/or polypeptides to cells, both in vitro and in vivo, such as expression vectors. These methods and compositions can be generally divided into two classes: viral delivery systems and non-viral delivery systems. Such methods are well known in the art and are readily adapted for use with the methods and compositions described herein. Such compositions and methods can be used to transfect or transduce cells in vitro or in vivo, such as to generate cell lines that express and, preferably, secrete the encoded chimeric polypeptide, or to therapeutically deliver nucleic acids to a subject. The nucleic acid components described herein are typically operably linked in frame to encode a fusion protein.

[154] As used herein, plasmids or viral vectors are agents that transfer the described nucleic acids into a cell without destroying it and contain a promoter that provides for expression of the nucleic acid molecule and/or polypeptide in the cells into which they have been delivered. Viral vectors include, for example, adenovirus, adeno-associated virus, herpes virus, vaccinia virus, poliovirus, Sindbis virus and other RNA viruses, including HIV backbone viruses. Also preferred are any families of viruses that have properties of those viruses that make them suitable for use as vectors. Retroviral vectors are generally described in Coffin et al., Retroviruses, Cold Spring Harbor Laboratory Press (1997), which is incorporated herein by reference with respect to vectors and methods for their construction. Construction of replication-defective adenoviruses has been described (Berkner et al., J. Virol. 61:1213–20 (1987); Massie et al., Mol. Cell. Biol. 6:2872–83 (1986); Haj-Ahmad et al., J. Virol. 57:267–74 (1986); Davidson et al., J. Virol. 61:1226–39 (1987); Zhang et al., BioTechniques 15:868–72 (1993)). An advantage of using these viruses as vectors is that they are limited in the extent to which they can spread to other cell types, that is, they can replicate within the original infected cell but are unable to form new infectious virus particles. Recombinant viruses have been shown to provide high efficacy following direct in vivo delivery to respiratory epithelium, hepatocytes, vascular endothelium, CNS parenchyma, and a variety of other tissue sites. Other useful systems include, for example, vectors based on replicating and host-restricted non-replicating vaccinia virus.

[155] The proposed polypeptides and/or nucleic acid molecules can be delivered via virus-like particles. Virus-like particles (VLPs) consist of viral proteins derived from the structural proteins of the virus. Methods for creating and using virus-like particles are described, for example, in Garcea and Gissmann, Current Opinion in Biotechnology 15:513-7 (2004).

[156] The proposed polypeptides can be delivered using subviral dense bodies (DBs). DBs transfer proteins into target cells via membrane fusion. Methods for the creation and use of DBs are described, for example, in Pepperl-Klindworth et al., Gene Therapy 10:278–84 (2003).

[157] The proposed polypeptides can be delivered via tegument aggregates. Methods for creating and using tegument aggregates are described in International Publication No. WO 2006/110728.

[158] Non-viral delivery methods may include the use of expression vectors comprising nucleic acid molecules and nucleic acid sequences encoding polypeptides, the nucleic acids being operably linked to an expression regulatory sequence. Suitable vector backbones include, for example, those commonly used in the art, such as plasmids, artificial chromosomes, BACs, YACs, or PACs. A number of vectors and expression systems are commercially available from corporations such as Novagen (Madison, Wis.), Clonetech (Pal Alto, Calif.), Stratagene (La Jolla, Calif.), and Invitrogen/Life Technologies (Carlsbad, Calif.), the vectors typically comprising one or more regulatory regions. Regulatory regions include, but are not limited to, promoter sequences, enhancer sequences, response elements, protein recognition sites, inducible elements, protein binding sequences, 5' and 3' untranslated regions (UTRs), transcription initiation sites, termination sequences, polyadenylation sequences, and introns. Such vectors can also be used to generate chimeric polypeptides by expression in a suitable host cell, such as CHO cells.

[159] Preferred promoters regulating transcription from vectors in mammalian host cells may be obtained from a variety of sources, such as genomes of viruses such as polyoma virus, simian virus 40 (SV40), adenovirus, retrovirus, hepatitis B virus and, most preferably, cytomegalovirus (CMV), or from heterologous mammalian promoters, such as the β-actin promoter or the EF1α promoter, or from hybrid chimeric promoters (e.g., the CMV promoter fused to the β-actin promoter). Of course, promoters from the host cell or related species are also suitable in the context of this document.

[160] An enhancer generally refers to a DNA sequence that does not function at any fixed distance from the transcription initiation site and may be either 5' or 3' relative to the transcription unit. Additionally, enhancers may be located within an intron as well as within the coding sequence itself. They are typically 10 to 300 base pairs (bp) in length and function in cis. The function of an enhancer is usually to increase transcription from adjacent promoters. Enhancers may also contain response elements that mediate transcriptional regulation. Although many enhancer sequences are known from mammalian genes (globin, elastase, albumin, fetoprotein, and insulin), an enhancer from a eukaryotic virus is typically used for general expression. Preferred examples are the SV40 enhancer on the late side of the replication origin, the cytomegalovirus early promoter enhancer, the polyoma virus enhancer on the late side of the replication origin, and adenovirus enhancers.

[161] The promoter and/or enhancer may be inducible (e.g., chemically or physically regulated). A chemically regulated promoter and/or enhancer may, for example, be regulated by the presence of an alcohol, tetracycline, steroid, or metal. A physically regulated promoter and/or enhancer may, for example, be regulated by environmental factors such as temperature and light. Optionally, the promoter and/or enhancer region may act as a constitutive promoter and/or enhancer to maximize expression of the region of the transcription unit dedicated to transcription. In certain vectors, the promoter and/or enhancer region may be active in a cell type-specific manner. Optionally, in certain vectors, the promoter and/or enhancer region may be active in eukaryotic cells, regardless of the cell type. Preferred promoters of this type are the CMV promoter, SV40 promoter, β-actin promoter, EF1α promoter and the retroviral long terminal repeat (LTR).

[162] Vectors may also contain, for example, origins of replication and/or markers. A marker gene may confer a selectable phenotype on a cell, such as resistance to an antibiotic. The marker product is used to determine whether the vector has been delivered to the cell and whether it is expressed after delivery. Examples of selectable markers for mammalian cells include dihydrofolate reductase (DHFR), thymidine kinase, neomycin, neomycin analog G418, hygromycin, puromycin, and blasticidin. If such selectable markers are successfully transferred into a mammalian host cell, the transformed mammalian host cell is able to survive when placed under selective pressure. Examples of other markers include, for example, the E. coli lacZ gene, green fluorescent protein (GFP), and luciferase. Additionally, the expression vector may contain a tag sequence designed to facilitate detection (e.g., purification or localization) of the expressed polypeptide. Tag sequences, such as GFP, glutathione S-transferase (GST), polyhistidine, c-myc, hemagglutinin, or FLAG™ tags (Kodak; New Haven, Conn.), are typically expressed as a fusion to the encoded polypeptide. Such tags may be inserted anywhere in the polypeptide, including at the carboxy- or amino-terminus.

[163] As used herein, the terms peptide, polypeptide or protein are used broadly to refer to two or more amino acids linked by a peptide bond. Protein, peptide and polypeptide are also used interchangeably herein to refer to amino acid sequences. It should be understood that the term polypeptide as used herein does not imply a particular size or number of amino acids comprising the molecule and that a peptide of the invention may comprise up to several amino acid residues or more. As used herein, a subject may be a vertebrate, more particularly a mammal (e.g., a human, horse, cat, dog, cow, pig, sheep, goat, mouse, rabbit, rat and guinea pig), bird, reptile, amphibian, fish and any other animal. The term does not imply a particular age or gender. Thus, it is intended that adult and neonatal male and female subjects are covered. As used herein, the terms patient or subject may be used interchangeably and may refer to a subject with a disease or disorder (e.g., cancer). The term patient or subject includes humans and veterinary subjects.

[164] A subject at risk of developing a disease or disorder may be genetically predisposed to the disease or disorder, such as having a family history or having a mutation in a gene that causes the disease or disorder, or exhibiting early signs or symptoms of the disease or disorder. A subject who currently has a disease or disorder has one or more symptoms of the disease or disorder, and may be diagnosed with the disease or disorder.

[165] The methods and agents described herein are useful as both prophylactic and therapeutic treatment. In the case of prophylactic use, a therapeutically effective amount of the chimeric polypeptides or chimeric nucleic acid sequences encoding the chimeric polypeptides described herein is administered to the subject prior to onset (e.g., prior to the onset of obvious symptoms of cancer or inflammation) or during early onset (e.g., following the onset of initial signs and symptoms of cancer or inflammation). Prophylactic administration can last from several days to several years prior to the onset of symptoms of cancer or inflammation. Prophylactic treatment can be used, for example, in the preventive treatment of subjects who have been diagnosed with a genetic predisposition to cancer. Therapeutic treatment includes administering to the subject a therapeutically effective amount of the chimeric polypeptides or chimeric nucleic acid sequences encoding the chimeric polypeptides described herein after the diagnosis or development of cancer or inflammation (e.g., an autoimmune disease). Prophylactic use can also be used when a patient is undergoing treatment, such as chemotherapy, where inflammation is expected.

[166] In accordance with the methods described herein, an effective amount of an agent (e.g., a chimeric polypeptide) is administered to a subject. The terms effective amount and effective dosage are used interchangeably. The term effective amount is defined as any amount necessary to produce a desired physiological response. Effective amounts and regimens for administering the agent can be determined empirically, and such determinations are within the skill in the art. Dosage ranges for administration are large enough to result in the desired effect, wherein one or more symptoms of the disease or disorder are modified (e.g., reduced or slowed). The dosage should not be so large as to cause significant undesirable side effects, such as undesirable cross-reactions, anaphylactic reactions, etc. In general, the dosage will vary depending on age, pathological condition, sex, type of disease, extent of the disease or disorder, route of administration, or whether other drugs are included in the regimen, and can be determined by one of skill in the art. The dosage may be adjusted by the individual physician in the event of any contraindications. Dosages may vary and may be administered in one or more doses daily over one or more days. Guidance on appropriate dosages for given classes of pharmaceutical products can be found in the literature.

[167] As used herein, the terms treating or treating refer to a method of reducing the effect of a disease or pathological condition or a symptom of a disease or pathological condition. Thus, in the described method, treating may refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the severity of the stated disease or pathological condition or a symptom of the disease or pathological condition. For example, a method of treating a disease is considered a treatment if a 10% reduction in one or more symptoms of the disease in a subject is observed compared to a control. Thus, the reduction may be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percentage reduction between 10% and 100% compared to native or control levels. It should be understood that treatment does not necessarily refer to a cure or complete elimination of the disease, pathological condition or symptoms of the disease or pathological condition.

[168] As used herein, the terms prevent and preventing a disease or disorder refer to an action, such as administering a chimeric polypeptide or a nucleic acid sequence encoding a chimeric polypeptide, that occurs before or about the same time a subject begins to exhibit one or more symptoms of the disease or disorder that inhibits or slows the onset or worsening of one or more symptoms of the disease or disorder. As used herein, use of the terms reduce, decrease, or inhibit includes a change of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to a control level. Such terms may include, but do not necessarily include, complete elimination.

[169] IL-2 variants have been developed that are selective for IL2Rαβγ over IL2Rβγ (Shanafelt, A. B., et al., 2000, Nat Biotechnol. 18:1197–202; Cassell, D. J., et al., 2002, Curr Pharm Des., 8:2171–83). These variants have amino acid substitutions that reduce their affinity for IL2Rβ. Because IL-2 has undetectable affinity for IL2Rγ, these variants therefore have reduced affinity for the IL2Rβγ receptor complex and reduced ability to activate IL2Rβγ-expressing cells, but retain the ability to bind IL2Rα and the ability to bind and activate the IL2Rαβγ receptor complex.

[170] One of these variants, IL2/N88R (Bay 50-4798), has been clinically investigated as a low-toxicity version of IL-2 as an immune stimulator based on the hypothesis that IL2Rβγ-expressing NK cells are the major contributors to toxicity. Bay 50-4798 has been shown to selectively stimulate the proliferation of activated T cells over NK cells and has been evaluated in phase 1/11 clinical trials in cancer patients (Margolin, K., et al., 2007, din Cancer Res., 13:3312-9) and HIV patients (Davey, R. T., et al., 2008, J Interferon Cytokine Res., 28:89-100). These clinical studies showed that Bay 50-4798 was significantly safer and more tolerable than aldesleukin, and also showed that it increased levels of CD4+CD25+ T cells, a cell population enriched in Treg cells. Since these trials, research in the field has further defined the identity of Treg cells and demonstrated that Treg cells selectively express IL2Rapy (reviewed in Maiek, T.R., et al., 2010, Immunity, 33:153-65).

[171] In addition, mutants can be generated that selectively alter affinity for the CD25 chain compared to native IL-2.

[172] IL-2 can be engineered to produce mutants that bind the IL-2R complex as a whole or the IL-2Rα subunit in particular with an affinity that differs from the corresponding wild-type IL-2 or a currently available mutant (called C125S because the cysteine at position 125 is replaced by a serine residue).

[173] Accordingly, the present invention relates to mutant interleukin-2 (IL-2*) polypeptides that comprise an amino acid sequence that is at least 80% identical to wild-type IL-2 (e.g., 85, 87, 90, 95, 97, 98, or 99% identical) and that binds, relative to WT IL-2, with greater affinity to a trimeric IL-2 receptor than to a dimeric IL-2 receptor. Typically, the muteins also bind the IL-2 receptor α-subunit (IL-2Rα) with an affinity that exceeds the affinity with which wild-type IL-2 binds IL-2Rα. The amino acid sequence within the mutant IL-2 polypeptides may vary from SEQ ID NO: 1 (UniProtKB Accession Number P60568) by containing (or only containing) one or more amino acid substitutions, which may be considered conservative or non-conservative substitutions. Non-naturally occurring amino acids may also be included. Alternatively, the amino acid sequence may vary from SEQ ID NO: 1 (which may be considered the "reference sequence") by containing and adding and/or deleting one or more amino acid residues. In particular, the amino acid sequence may differ from SEQ ID NO: 1 due to a mutation in at least one of the following positions of SEQ ID NO: 1: 1, 4, 8, 9, 10, 11, 13, 15, 26, 29, 30, 31, 35, 37, 46, 48, 49, 54, 61, 64, 67, 68, 69, 71, 73, 74, 75, 76, 79, 88, 89, 90, 92, 99, 101, 103, 114, 125, 128 or 133 (or combinations thereof). As indicated, at least one of these positions may be altered, as may two, three, four, five, six, seven, eight, nine, ten, or 11 or more (up to all) of the positions. For example, the amino acid sequence may differ from SEQ ID NO: 1 at positions 69 and 74 and additionally at one or more of positions 30, 35, and 128. The amino acid sequence may also differ from SEQ ID NO: 2 (described in U.S. Pat. No. 7,569,215, incorporated herein by reference) at one of the following groups of positions: (a) positions 64, 69, and 74; (b) positions 69, 74, and 101; (c) positions 69, 74, and 128; (d) positions 30, 69, 74, and 103; (e) positions 49, 69, 73, and 76; (f) items 69, 74, 101 and 133; (g) items 30, 69, 74 and 128; (h) items 69, 74, 88 and 99; (i) items 30, 69, 74 and 128; (j) items 9, 11, 35, 69 and 74; (k) items 1, 46, 49, 61, 69 and 79; (l) items 48, 68, 71, 90, 103 and 114; (m) items 4, 10, 11, 69, 74, 88 and 133; (n) items 15, 30, 31, 35, 48, 69, 74 and 92; (o) positions 30, 68, 69, 71, 74, 75, 76 and 90; (p) positions 30, 31, 37, 69, 73, 74, 79 and 128; (q) positions 26, 29, 30, 54, 67, 69, 74 and 92; (r) positions 8, 13, 26, 30, 35, 37, 69, 74 and 92; and (s) positions 29, 31, 35, 37, 48, 69, 71, 74, 88 and 89. Other than mutations at these positions, the amino acid sequence of the mutant IL-2 polypeptide may otherwise be identical to SEQ ID NO: 1. With respect to specific substitutions, the amino acid sequence may differ from SEQ ID NO: 1 by the presence of one or more of the following mutations: A1T, S4P, K8R, K9T, T10A, Q11R, Q13R, E15K, N26D, N29S, N30S, N30D, N30T, Y31H, Y31C, K35R, T37A, T37R, M46L, K48E, K49R, K49E, K54R, E61D, K64R, E67G, E68D, V69A, N71T, N71A, N71R, A73V, Q74P, S75P, K76E, K76R, H79R, N88D, I89V, N90H, I92T, S99P, T101A, F103S, I114V, I128T, I128A, T133A, or T133N. Our nomenclature follows that accepted in the scientific literature, in which the one-letter code for an amino acid in the wild-type or reference sequence is followed by its position in the sequence, and then the one-letter code for the amino acid with which it is substituted. Thus, A1T denotes the replacement of an alanine residue at position 1 with threonine. Other mutant polypeptides within the scope of the present invention include those comprising a mutant of SEQ ID NO: 2 having substitutions at V69 (e.g., A) and Q74 (e.g., P). For example, the amino acid sequence may comprise one of the following groups of mutations compared to SEQ ID NO: 2: (a) K64R, V69A and Q74P; (b) V69A, Q74P and T101A; (c) V69A, Q74P and I128T; (d) N30D, V69A, Q74P and F103S; (e) K49E, V69A, A73V and K76E; (f) V69A, Q74P, T101A and T133N; (g) N30S, V69A, Q74P and I128A; (h) V69A, Q74P, N88D and S99P; (i) N30S, V69A, Q74P and I128T; (j) K9T, Q11R, K35R, V69A and Q74P; (k) A1T, M46L, K49R, E61D, V69A and H79R; (l) K48E, E68D, N71T, N90H, F103S and I114V; (m) S4P, T10A, Q11R, V69A, Q74P, N88D and T133A; (n) E15K, N30S Y31H, K35R, K48E, V69A, Q74P and I92T; (o) N30S, E68D, V69A, N71A, Q74P, S75P, K76R and N90H; (p) N30S, Y31C, T37A, V69A, A73V, Q74P, H79R and I128T; (q) N26D, N29S, N30S, K54R, E67G, V69A, Q74P and I92T; (r) K8R, Q13R, N26D, N30T, K35R, T37R, V69A, Q74P and I92T; and (s) N29S, Y31H, K35R, T37A, K48E, V69A, N71R, Q74P, N88D and I89V. SEQ ID NO: 2 is described in U.S. Patent No. 7,569,215, which is incorporated herein by reference as an exemplary IL-2 polypeptide sequence that can be used in the present invention.

[174] As noted above, any of the mutant IL-2 polypeptides described herein may comprise the sequences described; they may also be limited to sequences described by or otherwise identical to SEQ ID NO: 1. In addition, any of the mutant IL-2 polypeptides described herein may optionally comprise a substitution of the cysteine residue at position 125 with another residue (e.g., serine) and/or may optionally comprise a deletion of the alanine residue at position 1 of SEQ ID NO: 1.

[175] The mutant IL-2 polypeptides described herein can bind to the IL-2Rα subunit with a K _d of less than about 28 nM (e.g., less than about 25 nM; less than about 5 nM; about 1 nM; less than about 500 pM; or less than about 100 pM). In particular, the mutant IL-2 polypeptide can have an equilibrium affinity constant of less than 1.0 nM (e.g., about 0.8, 0.6, 0.4, or 0.2 nM). Affinity can also be expressed as a relative rate of dissociation from the IL-2Rα subunit or from an IL-2 receptor complex (e.g., a complex expressed on the cell surface or otherwise membrane-associated). For example, mutant IL-2 polypeptides can dissociate from, for example, IL-2Rα, at a reduced rate compared to a wild-type polypeptide or an IL-2 therapeutic, for example, IL-2*. Alternatively, affinity can be characterized as the time or average time that an IL-2* polypeptide is present, for example, on the surface of a cell expressing IL-2R. For example, an IL-2* polypeptide can be present on a receptor for a time that is at least about 2, 5, 10, 50, 100, or 250 times (or more) longer.

[176] Materials, compositions, and components that can be used for, can be used in combination with, can be used in the preparation of, or that are products of the disclosed methods and compositions are described. These and other materials are described herein; it is understood that where combinations, subsets, interactions, groups, etc. of these materials are described, although specific reference to each of the various individual and collective combinations and permutations of these compounds may not be explicitly described, each is separately contemplated and described herein. For example, where a method is described and discussed and a number of modifications that can be made to a number of molecules, including the method, are discussed, each and every combination and permutation of the method and the modifications that are possible are explicitly contemplated unless otherwise explicitly indicated. Likewise, any subset or combination thereof is also explicitly contemplated and described. This concept applies to all aspects of the invention, including, but not limited to, steps in methods using the disclosed compositions. Thus, while there are a number of additional steps that can be performed, it should be understood that each of these additional steps can be performed using any specific method steps or combination of method steps of the described methods, and that each such combination or subset of combinations is expressly contemplated and considered to be described.

[177] The publications cited in this document and the materials in relation to which they are cited are expressly incorporated herein by reference in their entirety.

EXAMPLES

[178] The following are examples of methods and compositions according to the present invention. It should be understood that various other embodiments can be practiced taking into account the description provided herein.

Example 1: Detection of IL-2, IL-2 mutein, IL-2Rα and IL-2Rγ in fusion proteins by ELISA

[179] Detection of the IL-2 mutein is performed using a commercially available antibody, such as anti-1b-2 monoclonal antibody (JES6-1A12) (BD Pharmingen; San Jose, Calif.). A positive control is used to demonstrate whether the monoclonal antibody recognizes the cytokine or mutein. Antibodies against the IL-2Rα and IL-2Rγ chains can also be used. Wells of a 96-well plate are coated with antibody (2.5 μg/ml) in PBS. Wells are blocked with 5% nonfat milk in PBS containing 0.2% Tween®20 (PBS-M-Tw), and fusion proteins are added for 12 hours at 37°C. After washing, an anti-IL-2 biotin-labeled antibody, such as JES5H4 (BD Pharmingen), was added and binding was detected using streptavidin-HRP (Southern Biotechnology Associates; Birmingham, Ala.). The ELISA plate reaction was initiated by adding 50 μl of O-phenylenediamine (OPD) (Sigma-Aldrich) in 0.1 M citrate, pH 4.5, and 0.04% _H2O2 , stopped by adding 50 μl/well _of 2N _{H2SO4 ,} and the absorbance was read at 490 nm.

Example 2: Cleavage of the fusion protein by MMP9 protease

[180] One skilled in the art will be familiar with methods for performing a protein cleavage assay. 100 μg of protein in 1xPBS, pH 7.4, was cleaved with 1 μg of active MMP9 (Sigma catalog #SAE0078-50 or Enzo catalog #BML-SE360) and incubated at room temperature for up to 16 hours. The cleaved protein was then used in a functional assay or stored at -80°C until analyzed. The extent of cleavage was monitored by SDS-PAGE using methods known in the art. As illustrated in Fig. 10, complete cleavage of the fusion proteins by the MMP9 protease was observed.

Example 3: CTLL-2 Analysis

[181] CTLL2 cells (ATCC) were plated in suspension at 500,000 cells/well in culture medium with or without 40 mg/ml human serum albumin (HSA) and stimulated with serial dilutions of recombinant hIL2 or activatable hIL2 for 72 h at 37°C and 5% CO ₂ . The activity of uncleaved and cleaved activatable hIL2 was assayed. Cleaved activatable hIL2 was obtained by incubation with active MMP9. Cell activity was assessed using the CellTiter-Glo luminescent cell viability assay (Promega). The results are shown in Figs. 7–9.

Example 4: Protease cleavage of the IL-1/IL-1Rα/IL-1Rγ chimeric polypeptide results in increased antibody availability and biologically active IL-2 mutein

[182] Biochemical characterization of IL-2 mutein fusion proteins was performed before and after digestion with a protease such as PSA. Immunoblot analysis showed that the fusion proteins could be cleaved by PSA and that there was an increase in the predicted small molecular weight cleavage product of approximately 20 kDa that reacted with anti-IL-2 antibody after treatment of the samples with PSA. The extent of cleavage was dependent on the amount of PSA as well as the incubation time. Interestingly, when the fusion protein was analyzed before and after PSA digestion by ELISA, the apparent amount of IL-2 was found to be increased after PSA digestion. In this experiment, there was an approximately 2- to 4-fold increase in the apparent amount of IL-2 detected by sandwich ELISA depending on the design, suggesting that antibody binding was partially hindered in the intact fusion protein. Aliquots of the same samples were also assayed after PSA digestion using the CTLL-2 cell line, which requires IL-2 for growth and survival, and whose cell viability can be verified using the colorimetric MTT assay. In this assay, the more dilute the supernatant, the more biologically active IL-2 it contains; and an increase in biologically active IL-2 was observed after PSA digestion. The increase in IL-2 mutein would suggest that after PSA digestion there is an increase in the predicted small molecular weight cleavage fragment of approximately 20 kDa that is reactive with anti-IL-2 antibody, increased accessibility to the antibody, and, most importantly, an increase in biologically active IL-2 mutein.

Example 5. In vivo delivery of protease-activated fusion protein results in decreased tumor growth

[183] The fusion polypeptides have been studied to determine whether they can exert biological effects in vivo. These experiments utilize a system in which tumor cells injected intraperitoneally rapidly and preferentially attach to and grow primarily on milky spots, a series of organized immune aggregates found in the omentum (Gerber et al., Am. J. Pathol. 169:1739–52 (2006)). This system offers a convenient way to study the effects of fusion protein treatment on tumor growth, since fusion proteins can be repeatedly delivered intraperitoneally and tumor growth can be analyzed by studying dissociated omental cells. The colon 38 cell line, a rapidly growing tumor cell line that expresses both MMP2 and MMP9 in vitro, can be used for these experiments. Omental tissue normally expresses relatively low levels of MMP2 and MMP9, but when colon 38 tumor is present on the omentum, MMP levels are elevated. Using this tumor model, the ability of IL-2 mutein fusion proteins to influence tumor growth is examined. Colon 38 cells are injected intraperitoneally, allowed to attach and grow for 1 day, and then treated intraperitoneally with the fusion protein daily. On day 7, animals are sacrificed and the omenta are examined for tumor growth using flow cytometry and colony formation assays.

Example 6: Construction of a generic IL2 activation protein targeting CD20 Generation of the IL2 activation domain.

[184] An IL-2 polypeptide capable of binding to a CD20 polypeptide present in a tumor or on a tumor cell is prepared as follows. A nucleic acid is prepared that comprises nucleic acid sequences: (1) encoding an IL-2 polypeptide sequence and (2) one or more polypeptide linkers. The plasmid constructs of the activatable interleukin may have optional Flag, His, or other affinity tags and are electroporated into HEK293 or other suitable human or mammalian cell lines and purified. A validation assay includes a T cell activation assay using T cells susceptible to IL-2 stimulation in the presence of protease.

Construction of CD20-binding domain scFv

[185] CD20 is one of the cell surface proteins present on B lymphocytes. The CD20 antigen is found on normal and malignant pre-B and mature B lymphocytes, including more than 90% of non-Hodgkin lymphoma (NHL) B cells. This antigen is absent from hematopoietic stem cells, activated B lymphocytes (plasma cells), and normal tissue. Several antibodies, mainly of murine origin, have been described in this regard: 1F5, 2B8/C2B8, 2H7, and 1H4.

[186] Therefore, human or humanized anti-CD20 antibodies are used to generate scFv sequences for the CD20 binding domains of the activated interleukin protein. DNA sequences encoding human or humanized VL and VH domains are obtained, and the constructs are optionally codon optimized for expression in cells from Homo sapiens. The order in which the VL and VH domains are present in the scFv is varied (i.e., VL-VH or VH-VL orientation), and three copies of "G4S" or the "G4S" subunit ( _G4S ) ₃ link the variable domains to create the scFv domain. The anti-CD20 scFv plasmid constructs may have optional Flag, His, or other affinity tags, and are electroporated into HEK293 or other suitable human or mammalian cell lines and purified. Validation analysis included FACS binding analysis, Proteon kinetic analysis, and staining of CD20-expressing cells.

Cloning of DNA expression constructs encoding the activated IL2 protein

[187] The IL2 activatable construct with protease cleavage site domains is used to construct the activatable interleukin protein in combination with an anti-CD20 scFv domain and a serum half-life extender element (e.g., HSA binding peptide or VH domain). To express the activatable interleukin protein in CHO cells, the coding sequences of all protein domains are cloned into a mammalian expression vector system. Briefly, the gene sequences encoding the activatable interleukin protein domain, the serum half-life extender element, and the CD20 binding domain along with the L1 and L2 peptide linkers are synthesized separately and subcloned. The resulting constructs were ligated together in the order CD20 binding domain - L1 - IL2 subunit 1 - L2 - protease cleavage domain - L3 - IL2 subunit 2 - L4 - anti-CD20 scFv - L5 - serum half-life extender element to generate the final construct. All expression constructs contain coding sequences for an N-terminal signal peptide and a C-terminal hexahistidine (6xHis) tag to facilitate protein secretion and purification, respectively.

Expression of IL2-activated proteins in stably transfected CHO cells

[188] Using a CHO cell expression system (F1p-In®, Life Technologies), a derivative of Chinese hamster ovary cells CHO-K1 (ATCC, CCL-61) (Kao and Puck, Proc. Natl. Acad Sci USA 1968;60(4):1275-81), adherent cells were subcultured according to standard cell culture protocols provided by Life Technologies.

[189] To adapt to growth in suspension, cells are separated from tissue culture tubes and placed in serum-free medium. Suspension-adapted cells are cryopreserved in medium containing 10% DMSO.

[190] Recombinant CHO cell lines stably expressing secreted activatable interleukin proteins are generated by transfection of suspension-adapted cells. Viable cell density is measured twice weekly during selection with the antibiotic hygromycin B, and cells are centrifuged and resuspended in fresh selection medium at a maximum density of 0.1×10 ⁶ viable cells/mL. Pools of cells stably expressing activatable interleukin proteins are recovered after 23 weeks of selection, at which time the cells are transferred to standard culture medium in shake tubes. Expression of recombinant secreted proteins is confirmed by gel electrophoresis or flow cytometry. Stable cell pools are cryopreserved in DMSO-containing medium.

[191] Activated IL2 proteins are produced in 10-day fed-fed cultures of stably transfected CHO cell lines by secretion into cell culture supernatant. Cell culture supernatants are harvested after 10 days when culture viability is typically >75%. Samples are taken from production cultures after 24 hours and assessed for cell viability and density. On the day of harvest, cell culture supernatants are clarified by centrifugation and vacuum filtration before further use.

[192] Protein expression titers and product integrity in cell culture supernatants were analyzed by SDS-PAGE.

Purification of IL2-activated proteins

[193] Activated IL2 proteins are purified from CHO cell culture supernatants using a two-step procedure. In the first step, constructs are subjected to affinity chromatography, followed by size exclusion chromatography (SEC) on Superdex 200 in the second step. Samples are buffer exchanged and concentrated by ultrafiltration to a typical concentration of >1 mg/mL. Purity and homogeneity (typically >90%) of the final samples are assessed by SDS-PAGE under reducing and non-reducing conditions, followed by immunoblotting with anti-HSA or anti-idiotype antibody and analytical SEC, respectively. Purified proteins are stored in aliquots at -80°C until use.

Example 7: Determination of antigen affinity by flow cytometry

[194] The IL2 upregulated proteins from Example 6 were tested for binding affinity to human CD20 ⁺ cells and cynomolgus monkey CD20 ⁺ cells.

[195] CD20 ⁺ cells were incubated with 100 μl of the serial dilutions of the activatable interleukin proteins from Example 6 and at least one protease. After washing three times with FACS buffer, the cells were incubated with 10 μg/ml mouse monoclonal anti-idiotype antibody in the same buffer for 45 min on ice. After a second round of washing, the cells were incubated with 10 μg/ml FITC-conjugated goat anti-mouse IgG under the same conditions as before. As a control, the cells were incubated with anti-His IgG, followed by FITC-conjugated goat anti-mouse IgG without the activatable IL2 proteins. The cells were washed again and resuspended in 0.2 ml FACS buffer containing 2 μg/ml propidium iodide (PI) to exclude dead cells. Fluorescence of 1× ¹⁰⁴ living cells was measured using a Beckman-Coulter FC500 MPL flow cytometer and MXP software (Beckman-Coulter, Krefeld, Germany) or a Millipore Guava EasyCyte flow cytometer and Incyte software (Merck Millipore, Schwalbach, Germany). The mean fluorescence intensity of the cell samples was calculated using CXP software (Beckman-Coulter, Krefeld, Germany) or Incyte software (Merck Millipore, Schwalbach, Germany). After subtracting the fluorescence intensity values of cells stained with secondary and tertiary reagents only, these values are then used to calculate k _d values using the single-site binding equation (hyperbola) in GraphPad Prism (version 6.00 for Windows, GraphPad Software, La Jolla California USA).

[196] CD20 binding and cross-reactivity were assessed in human CD20 ⁺ tumor cell lines. The k _d ratio of cross-reactivity was calculated using k _d values determined in CHO cell lines expressing recombinant human or recombinant cynomolgus antigens.

Example 8: Cytotoxicity Assay

[197] The IL2 upregulated protein from Example 6 was assessed in vitro for its mediation of the immune response to target CD20 ⁺ cells.

[198] Fluorescently labeled CD20 ⁺ REC-1 cells (a mantle cell lymphoma cell line, ATCC CRL-3004) are incubated with isolated PBMCs from random donors or CD 15 T cells (a standardized T cell line) as effector cells in the presence of the IL2-activatable protein from Example 6 and at least one protease. After incubation for 4 h at 37°C in a humidified incubator, the release of fluorescent dye from the target cells into the supernatant is determined in a spectrofluorometer. Target cells that were incubated without the IL2-activatable protein from Example 6 and target cells completely lysed by adding saponin at the end of the incubation serve as negative and positive controls, respectively.

[199] Based on the measured remaining viable target cells, the percentage of specific cell lysis was calculated according to the following formula: [1-(number of viable targets (sample)/number of viable targets (spontaneous))] × 100%. Sigmoidal dose-response curves and _EC50 values were calculated using nonlinear regression/4-parameter logistic fitting using GraphPad software. The lysis values obtained for a given antibody concentration were used to calculate sigmoidal dose-response curves using 4-parameter logistic fitting analysis using Prism software.

Example 9: Pharmacokinetics of IL2-activated proteins

[200] The activated IL2 protein from Example 6 was evaluated for half-life in animal studies.

[201] IL2 AP is administered to cynomolgus monkeys as a 0.5 mg/kg bolus injection into a subcutaneous vein. Another group of cynomolgus monkeys receives an IL2 construct of comparable size but lacking the serum half-life extender element. The third and fourth groups receive an IL2 construct with a serum half-life extender element and a CD20 cytokine and a serum half-life extender element, respectively, both comparable in size to IL2 AP. Each study group consists of 5 monkeys. Serum samples are obtained at the indicated time points, serially diluted, and protein concentrations are determined using a CD20 binding ELISA.

[202] Pharmacokinetic analyses are performed using plasma concentrations of the test drug. Group mean plasma data for each test drug are fitted to a multiexponential profile when plotted against time after dosing. The data are fitted to a standard two-compartment model with bolus administration and first-order rate constants for the distribution and elimination phases. The general equation for the best fit to the IV data is: c(t) = Ae ^{- αt} + Be ^{- βt} , where c(t) is the plasma concentration at time t, A and B are the y-intercepts, and α and β are the apparent first-order rate constants for the distribution and elimination phases, respectively. The α phase represents the initial elimination phase and reflects the distribution of the protein throughout the animal's extracellular fluid, while the second part, or β phase, of the decline curve represents the true plasma elimination. Methods for approximating such equations are well known in the art. For example, A=D/V(α-k21)/(α-β), B=D/V(β-k21)/(α-β), and α and β (for α>β) are the roots of the quadratic equation: ^r2 +(k12+k21+k10)r+k21k10=0 using the estimated parameters V is the volume of distribution, k10 is the elimination rate, k12 is the rate of transfer from compartment 1 to compartment 2, and k21 is the rate of transfer from compartment 2 to compartment 1, and D is the administered dose.

[203] Data analysis: Concentration-time profiles were plotted using KaleidaGraph (KaleidaGraph™ V. 3.09 Copyright 1986-1997. Synergy Software. Reading, Pa.). Values reported as “less than required to report” (LNR) were not included in the PK analysis and were not presented graphically. Pharmacokinetic parameters were determined by compartmental analysis using WinNonlin software (WinNonlin® Professional V. 3.1 WinNonlin™ Copyright 1998-1999. Pharsight Corporation. Mountain View, Calif.). Pharmacokinetic parameters were calculated as described by Ritschel WA and Keams GL, 1999, in: Handbook Of Basic Pharmacokinetics Including Clinical Applications, 5 ^th edition, American Pharmaceutical Assoc., Washington, DC

[204] The activated IL2 protein of Example 6 is expected to have improved pharmacokinetic parameters, such as increased half-life, compared to proteins that lack the element to prolong serum half-life.

Example 10: Tumor xenograft model

[205] The IL2 upregulated protein from Example 6 is evaluated in a xenograft model.

[206] Female immunodeficient NOD/scid mice are irradiated with a sublethal dose (2 Gy) and inoculated subcutaneously with 4 × 10 ⁶ Ramos RA1 cells in the upper right flank. When tumors reach 100-200 mm ³ , animals are divided into 3 treatment groups. Groups 2 and 3 (8 animals each) are injected intraperitoneally with 1.5 × 10 ⁷ activated human T cells. Three days later, animals in Group 3 are sequentially treated with a total of 9 intravenous doses of 50 μg of the activatable interleukin protein from Example 1 (once daily for 9 sec). Groups 1 and 2 are treated with vehicle only. Body weight and tumor volume are determined over 30 days.

[207] Animals treated with the IL2 upregulated protein from Example 6 are expected to have a statistically significant reduction in tumor growth compared to the corresponding vehicle-treated control group.

[208] Although preferred embodiments of the present invention have been illustrated and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now become apparent to those skilled in the art without departing from the scope of the invention. It will be understood that various alternatives to the embodiments of the invention described herein may be used in practicing the invention. It is intended that the following claims define the scope of the invention and that they cover methods and structures within the scope of these claims, as well as equivalents thereof.

Example 11: HEK Blue Analysis

[209] HEK-Blue IL2 cells (InvivoGen) were plated in suspension at 50,000 cells/well in culture medium with or without 15-40 mg/ml human serum albumin (HSA) and stimulated with serial dilutions of recombinant hIL2 or activatable hIL2 for 24 h at 37°C and 5% CO ₂ . The activity of uncleaved and cleaved activatable hIL2 was assayed. Cleaved inducible hIL2 was obtained by incubation with active MMP9. IL12 activity was assessed by quantitative assessment of secreted alkaline phosphatase (SEAP) activity using the QUANTI-Blue reagent (InvivoGen), a colorimetric assay. The results are shown in Fig. 11.

Example 12: MS38 Experiments

[210] The MC38 cell line, a rapidly growing colon adenocarcinoma cell line that expresses MMP9 in vitro, was used to examine the ability of the fusion proteins to influence tumor growth using this tumor model.

Example 12a: MC38 IL-2POC Agents and Treatment:

Example 12b: MC38 IL-2 Agents and Treatment:

Example 12c: Processing ACP16, ACP132 and ACP21

Mice were anesthetized with isoflurane for cell implantation to reduce ulceration. CR female C57BL/6 mice were injected s.c. with 5 x 10 ⁵ MC38 tumor cells in 0% Matrigel in the flank. Cell injection volume was 0.1 ml/mouse. Mice were 8-12 weeks old at the start date. Pairs were matched when tumors reached a mean volume of 100-150 mm ³ and treatment was started. ACP16 was injected at 17, 55, 70, or 230 μg/animal; ACP132 was injected at 9, 28, 36, or 119 μg/animal; ACP21 was injected at 13, 42, 54, or 177 μg/animal. Body weight was measured at the start date and twice weekly thereafter until completion. Caliper measurements were made twice weekly until completion. Any adverse reactions were to be reported promptly. Any individual animal with a single observation of >30% body weight loss or three consecutive observations of >25% body weight loss was euthanized. In any group with a mean body weight loss of >20% or >10%, dosing was stopped; the group was not euthanized and allowed to recover. In groups with >20% weight loss, animals that reached the individual body weight loss endpoint were euthanized. If treatment-related body weight loss in a group recovered to 10% of baseline weight, dosing was resumed at a lower dose or with a less frequent dosing schedule. Exceptions for non-treatment-related % body weight recovery were allowed on a case-by-case basis. The endpoint was tumor growth inhibition (TGI). Animals were observed individually. The endpoint was a tumor volume of 1500 mm ³ or 45 days, whichever occurred first. The animals showing a response were observed for a longer period. When the final result was reached, the animals had to be sacrificed. The results are illustrated in Fig. 17.

Example 12d: MS38 repetitive stimulation

The cured mice (ACP16 treated) from Example 12b were restimulated with tumor implants to determine whether anti-tumor memory had formed after the initial treatment.

Agents and processing:

Procedures:

Mice were anesthetized with isoflurane for cell implantation to reduce ulceration. This portion of the study began on the day of implantation (day 1). Group 1 consisted of 33 CR female C57BL/6 mice that received 5 × 10 ⁵ MC38 tumor cells in 0% Matrigel subcutaneously in the flank. Groups 2–6 consisted of 33 CR female C57BL/6 mice that received 5 × 10 ⁵ MC38 tumor cells in 0% Matrigel subcutaneously in the flank. Tumors from the previous MC38 experiment (Example 12b) were implanted in the right flank of each animal. The cell injection volume was 0.1 ml/mouse. Control mice were 1417 weeks old at initiation. These mice were age-matched to the mice from the previous MC38 experiment (Example 12b). No active agent was administered during restimulation. Body weight was measured twice weekly until completion, as were caliper measurements. Any adverse reactions or death were reported promptly. Any individual animal with one observation of >30% body weight loss or three consecutive observations of >25% body weight loss was sacrificed. The endpoint was tumor growth retardation (TGR). Animals were observed individually. The endpoint of the experiment was a tumor volume of 1000 mm ^{for 3} or 45 days, whichever occurred first. When possible, responding animals were observed longer. Animals were sacrificed when the endpoint was reached. The results are shown in Fig. 15.

Example 13. Conditionally active fusion proteins that contain a blocking moiety that is a serum albumin binding moiety

[211] This example describes the production and activity of fusion proteins, preferably cytokines, that have inducible activity, i.e., they are inactive until induced, typically by separating a blocking moiety from an active moiety after cleavage of a linker between the blocking moiety and the active moiety. The fusion proteins comprise a single antibody variable domain (dAb) that binds serum albumin via the CDR loops and binds to the active moiety (in this case, anti-CD3 scFV) via one or more non-CDR loops (e.g., the C loop). The serum albumin binding blocking moiety is operably linked to the active moiety via a protease cleavable linker, and the active moiety is operably linked to the targeting domain (in this case an epidermal growth factor receptor (EGFR) dAb or an anti-prostate-specific membrane antigen (PSMA) dAb) via a linker that is not cleavable by protease. These fusion proteins can be administered as inactive proteins that are activated upon cleavage of the protease cleavable linker and subsequent release of the inhibitory albumin binding domain. The anti-CD3 scFV in the fusion proteins is a surrogate for the desired cytokine in the fusion proteins described in this invention. Similar fusion proteins that comprise a desired cytokine (e.g., IL-2, IL-12, interferon) or a functional fragment or mutein thereof, a targeting domain, and an albumin-binding dAb that also binds and inhibits the cytokine or a functional fragment or mutein thereof can be prepared using the methods described and illustrated herein. An anti-serum albumin dAb that binds and inhibits the activity of a desired cytokine or a functional fragment or mutein thereof can provide both steric masking of the cytokine (due to the proximity of the cytokine to bound serum albumin) and specific masking of the cytokine (due to binding to the cytokine via a loop other than the CDR (e.g., loop C)). An anti-serum albumin dAb that binds to and inhibits the activity of a desired cytokine or a functional fragment or mutein thereof can be generated using suitable methods, such as by introducing amino acid sequence diversity into non-CDR loops (e.g., loop C) of a binding anti-serum albumin dAb and screening for binding to the desired cytokine. Any suitable methods, such as phage display, can be used to perform the selection. For example, a typical anti-serum albumin dAb that can be used has the following sequence, and the amino acid sequence of loop C (shown in bold and underlined) can be diversified (e.g., randomized) and the resulting dAbs screened for binding to serum albumin via CDR interaction and to the cytokine via non-CDR loop interaction. If desired, the amino acid sequence of a known cytokine-binding peptide can be grafted into loop C.

A. Activation by ProTriTAC protease results in significantly increased activity in vitro

[212] Purified ProTriTAC (prodrug), non-cleavable ProTriTAC [prodrug (non-cleavable)], and a recombinant active drug fragment mimicking the activity of protease-activated ProTriTAC (active drug) were tested for binding to recombinant human CD3 in an ELISA assay, binding to purified human primary T cells in a flow cytometric assay, and functional efficacy in a T cell-dependent cellular cytotoxicity assay.

[213] For ELISA, soluble ProTriTAC proteins at the indicated concentrations were incubated with immobilized recombinant human CD3e (R&D Systems) for 1 h at room temperature in PBS supplemented with 15 mg/ml human serum albumin. Plates were blocked using SuperBlock (Thermo Fisher), washed with PBS containing 0.05% Tween-20, and detected using the non-competitive anti-CD3 idiotypic monoclonal antibody 11D3, followed by peroxidase-labeled secondary antibody and TMB-ELISA substrate solution (Thermo Fisher).

[214] For flow cytometry, soluble ProTriTAC proteins at the indicated concentrations were incubated with purified human primary T cells for 1 h at 4°C in the presence of PBS containing 2% fetal bovine serum and 15 mg/ml human serum albumin. Plates were washed with PBS containing 2% fetal bovine serum, detected using AlexaFluor 647-labeled non-competitive anti-CD3 idiotypic monoclonal antibody 11D3, and data analyzed using FlowJo 10 (FlowJo, LLC).

[215] To determine functional efficacy in a T cell-dependent cellular cytotoxicity assay, soluble ProTriTAC proteins at the indicated concentrations were incubated with purified resting human T cells (effector cells) and HCT116 cancer cells (target cells) at a 10:1 ratio of effector gap cells for 48 h at 37°C. The HCT116 target cell line was stably transfected with a luciferase reporter gene to allow measurement of T cell-mediated cell killing using ONE-Glo (Promega).

B. ProTriTAC exhibits potent, protease-dependent antitumor activity in a rodent tumor xenograft model

[216] ProTriTAC was evaluated for its in vivo antitumor activity in subcutaneous HCT116 xenograft tumor spiked with expanded human T cells in NCG immunodeficient mice. Specifically, 5× ¹⁰⁶ HCT116 cells were mixed with 2.5× ¹⁰⁶ expanded T cells per mouse on day 0. ProTriTAC was administered dosing every day × 10 by intraperitoneal injection beginning the following day. Tumor volume was determined using caliper measurements and calculated using the formula V = (length × width × width)/2 at the indicated time points.

C. Expression, purification and stability of exemplary ProTriTAC trispecific molecules

Protein production

[217] Sequences encoding the inducible fusion protein molecules were cloned into the mammalian expression vector pcDNA 3.4 (bivitrogen), preceded by a leader sequence and followed by a 6x histidine tag (SEQ ID NO: 136). Expi293F cells (Life Technologies A14527) were maintained in suspension in Optimal Growth Tubes (Thomson) at 0.2 to 8 × 1e6 cells/mL in Expi 293 medium. Purified plasmid DNA was transfected into Expi293 cells according to the Expi293 Expression System Kit (Life Technologies, A14635) protocols and maintained for 4-6 days post-transfection. Alternatively, sequences encoding fusion protein molecules were cloned into the mammalian expression vector pDEF38 (CMC ICOS), transfected into CHO-DG44 dhfr- cells, stable pools were generated, and cultured in production medium for up to 12 days before purification. The amount of representative fusion proteins in conditioned medium was estimated using an Octet RED 96 instrument with protein A tips (ForteBio/Pall) using a control fusion protein for the standard curve. Conditioned medium from any host cells was filtered and partially purified by affinity and desalting chromatography. Fusion proteins were then polished by ion exchange chromatography and, after pooling, stirred in neutral buffer containing excipients. Final purity was assessed by SDS-PAGE and analytical SEC using an Acquity BEH SEC 200 1.7 μm 4.6 x 150 mm column (Waters Corporation), resolved in an aqueous/organic mobile phase with excipients at neutral pH on a 1290 LC system, and peaks integrated using Chemstation CDS software (Agilent). Fusion proteins purified from CHO cells are shown in the SDS-PAGE shown below.

Stability assessment

[218] Purified fusion proteins in the two formulations were subaliquoted into sterile tubes and stressed by five freeze-thaw cycles, each comprising over 1 hour at -80°C and room temperature, or by incubation at 37°C for 1 week. Stressed samples were assessed for concentration and turbidity by UV spectrometry using UV-transparent 96-well plates (Corning 3635) with SpectraMax M2 and SoftMaxPro software (Molecular Devices), SDS-PAGE, and analytical SEC, and compared with the same analysis of unstressed control samples. An overlay of analytical SEC chromatograms of control and stressed samples for a single representative ProTriTAC molecule purified from 293 host cells is shown below.

[219] The results show that ProTriTACs were produced at comparable levels to regular TriTACs from stable CHO pools; and that the protein properties were stable after repeated freeze-thaw cycles and 37°C for 1 week.

D. Demonstration of functional masking and in vivo stability of ProTriTAC in a three-week pharmacokinetic study in cynomolgus monkeys

[220] A single dose of PSMA-targeted ProTriTAC (SEQ ID NO: 119), non-cleavable ProTriTAC (SEQ ID NO: 120), unmasked/non-cleavable TriTAC (SEQ ID NO: 123), and active drug mimicking protease-activated ProTriTAC (SEQ ID NO: 121) was administered to cynomolgus monkeys at 0.1 mg/kg by intravenous injection. Plasma samples were obtained at the indicated time points. ProTriTAC concentrations were determined using a ligand binding assay with biotinylated recombinant human PSMA (R&D systems) and sulfo-labeled anti-CD3 idiotype antibody 11D3 in an MSD assay (Meso Scale Diagnostic, LLC). Pharmacokinetic parameters were assessed using Phoenix WinNonlin pharmacokinetic software and a non-compartmental approach consistent with the intravenous bolus route of administration.

[221] To calculate the in vivo rate of prodrug conversion, the concentration of active drug in circulation was estimated by solving the following system of differential equations, where P is the prodrug concentration, A is the active drug concentration, k _a is the rate of prodrug activation in circulation, k _c,P is the rate of prodrug elimination, and k _c,A is the rate of active drug elimination.

[222] The clearance rates of prodrug, active drug, and non-cleavable control prodrug (k _c,NCLV ) were determined empirically in cynomolgus monkeys. To estimate the rate of prodrug activation in circulation, we assumed that the difference between the clearance rates of the cleavable prodrug and non-cleavable prodrug was due solely to non-specific activation in circulation. Therefore, the rate of conversion of prodrug to active drug in circulation was estimated by subtracting the clearance rate of the cleavable prodrug from the non-cleavable prodrug.

[223] The initial circulating concentration of the prodrug was determined empirically, and the initial concentration of the active drug was taken as zero.

Results and discussion

[224] The results of Example 13 show that it is possible to prepare fusion proteins that comprise a polypeptide with a desired therapeutic activity, such as a cytokine or a functional fragment or mutein thereof, or an anti-CD3 scFV, in which the therapeutic activity is masked by a masking domain that binds to both serum albumin and the active polypeptide. The masking domain is operably linked to the active domain via a protease cleavable linker. The results show that this type of fusion protein can be administered as an inactive protein that is activated upon cleavage by a protease at the desired site of therapeutic activity, such as a tumor.

[225] The amino acid sequences of the fusion proteins of Example 13 are provided as SEQ ID NOs: 116-123.

[226] Examples of fusion protein constructs are detailed in Table 3. In Table 3, "L" is an abbreviation for "linker" and "cleavable linker" is an abbreviation for "cleavable linker". Other abbreviations: "mIFNg" stands for mouse interferon gamma (IFNg); "hAlbumin" stands for human serum albumin (HSA); "mAlbumin" stands for mouse serum albumin.

INCLUSION BY LINK

[227] The entire contents of all patent and non-patent publications cited in this document are incorporated by reference in their entirety and for all purposes.

OTHER IMPLEMENTATION OPTIONS

[228] The foregoing description may cover many different inventions with independent application. Although each of these inventions has been described in preferred form(s), the specific embodiments described and illustrated herein are not to be taken in a limiting sense since many variations are possible. The subject matter of the present invention includes all novel and non-obvious combinations and subcombinations of the various elements, features, functions and/or properties described herein. In particular, the following claims point out certain combinations and subcombinations believed to be novel and non-obvious. Inventions embodied in other combinations and subcombinations of features, functions, elements and/or properties may be claimed in this application, in applications claiming priority to this application, or in related applications. Such claims that relate to another invention or to the same invention and have a broader, narrower, equivalent or different scope than the original claims are also considered to be included within the subject matter of this specification.

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SEQUENCE LIST

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Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr Cys Phe

580 585 590

Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala Leu Gly

595 600 605

Leu

<210> 3

<211> 8

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP7 cleavage domain sequence"

<400> 3

Lys Arg Ala Leu Gly Leu Pro Gly

1 5

<210> 4

<211> 40

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP7 cleavage domain sequence"

<400> 4

Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu Asp Glu

1 5 10 15

Arg Pro Leu Ala Leu Trp Arg Ser Asp Arg Asp Arg Asp Arg Asp Arg

20 25 30

Asp Arg Asp Arg Asp Arg Asp Arg

35 40

<210> 5

<211> 5

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP9 cleavage domain sequence"

<220>

<221> OPTION

<222> (3)..(3)

<223> /replacement="Thr"

<220>

<221> OPTION

<222> (4)..(4)

<223> /replacement="Ile"

<220>

<221> OPTION

<222> (5)..(5)

<223> /replacement="Thr"

<220>

<221> SITE

<222> (1)..(5)

<223> /note="Variant residues given in the sequence,

have no advantage over those in

designations for variant positions"

<400> 5

Pro Arg Ser Leu Ser

1 5

<210> 6

<211> 5

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP9 cleavage domain sequence"

<400> 6

Leu Glu Ala Thr Ala

1 5

<210> 7

<211> 10

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP11 cleavage domain sequence"

<400> 7

Gly Gly Ala Ala Asn Leu Val Arg Gly Gly

1 5 10

<210> 8

<211> 10

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP14 cleavage domain sequence"

<400> 8

Ser Gly Arg Ile Gly Phe Leu Arg Thr Ala

1 5 10

<210> 9

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP cleavage domain sequence"

<400> 9

Pro Leu Gly Leu Ala Gly

1 5

<210> 10

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP cleavage domain sequence"

<220>

<221> MOD_OST

<222> (6)..(6)

<223> Any amino acid

<400> 10

Pro Leu Gly Leu Ala Xaa

1 5

<210> 11

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP cleavage domain sequence"

<220>

<221> MOD_OST

<222> (4)..(4)

<223> Cys(me)

<400> 11

Pro Leu Gly Xaa Ala Gly

1 5

<210> 12

<211> 8

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP cleavage domain sequence"

<400> 12

Glu Ser Pro Ala Tyr Tyr Thr Ala

1 5

<210> 13

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP cleavage domain sequence"

<400> 13

Arg Leu Gln Leu Lys Leu

1 5

<210> 14

<211> 7

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP cleavage domain sequence"

<400> 14

Arg Leu Gln Leu Lys Ala Cys

1 5

<210> 15

<211> 7

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

MMP2, MMP9, MMP14 cleavage domain sequence"

<220>

<221> MOD_OST

<222> (3)..(3)

<223> Cit

<220>

<221> MOD_OST

<222> (5)..(5)

<223> Hof

<400> 15

Glu Pro Xaa Gly Xaa Tyr Leu

1 5

<210> 16

<211> 5

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Activator cleavage domain sequence

urokinase-type plasminogen (uPA)"

<400> 16

Ser Gly Arg Ser Ala

1 5

<210> 17

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Activator cleavage domain sequence

urokinase-type plasminogen (uPA)"

<400> 17

Asp Ala Phe Lys

1

<210> 18

<211> 5

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Activator cleavage domain sequence

urokinase-type plasminogen (uPA)"

<400> 18

Gly Gly Gly Arg Arg

1 5

<210> 19

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Sequence of the cleavage domain of the lysosomal enzyme"

<400> 19

Gly Phe Leu Gly

1

<210> 20

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Sequence of the cleavage domain of the lysosomal enzyme"

<400> 20

Ala Leu Ala Leu

1

<210> 21

<211> 2

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Sequence of the cleavage domain of the lysosomal enzyme"

<400> 21

Phe Lys

1

<210> 22

<211> 3

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Cathepsin B cleavage domain sequence"

<400> 22

Asn Leu Leu

1

<210> 23

<211> 5

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Cathepsin D" cleavage domain sequence

<220>

<221>MOD_OST

<222> (3)..(3)

<223> Cys(et)

<400> 23

About Ile Xaa Phe Phe

1 5

<210> 24

<211> 8

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Cathepsin K" cleavage domain sequence

<400> 24

Gly Gly Pro Arg Gly Leu Pro Gly

1 5

<210> 25

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Prostate-specific antigen cleavage domain sequence"

<400> 25

His Ser Ser Lys Leu Gln

1 5

<210> 26

<211> 7

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Prostate-specific antigen cleavage domain sequence"

<400> 26

His Ser Ser Lys Leu Gln Leu

1 5

<210> 27

<211> 9

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Prostate-specific antigen cleavage domain sequence"

<400> 27

His Ser Ser Lys Leu Gln Glu Asp Ala

1 5

<210> 28

<211> 10

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Sequence of the protease cleavage domain of the virus simplex

herpes"

<400> 28

Leu Val Leu Ala Ser Ser Ser Phe Gly Tyr

1 5 10

<210> 29

<211> 10

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Sequence of the HIV protease cleavage domain"

<400> 29

Gly Val Ser Gln Asn Tyr Pro Ile Val Gly

1 5 10

<210> 30

<211> 10

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Sequence of the CMV protease cleavage domain"

<400> 30

Gly Val Val Gln Ala Ser Cys Arg Leu Ala

1 5 10

<210> 31

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Thrombin cleavage domain sequence"

<220>

<221> MOD_OST

<222> (2)..(2)

<223> 2-carboxypiperidine

<400> 31

Phe Xaa Arg Ser

1

<210> 32

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Thrombin cleavage domain sequence"

<400> 32

Asp Pro Arg Ser Phe Leu

1 5

<210> 33

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Thrombin cleavage domain sequence"

<400> 33

Pro Pro Arg Ser Phe Leu

1 5

<210> 34

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Caspase-3 cleavage domain sequence"

<400> 34

Asp Glu Val Asp

1

<210> 35

<211> 5

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Caspase-3 cleavage domain sequence"

<400> 35

Asp Glu Val Asp Pro

1 5

<210> 36

<211> 8

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Caspase-3 cleavage domain sequence"

<400> 36

Lys Gly Ser Gly Asp Val Glu Gly

1 5

<210> 37

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Cleavage domain sequence

interleukin 1-beta-converting enzyme"

<400> 37

Gly Trp Glu His Asp Gly

1 5

<210> 38

<211> 7

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Enterokinase cleavage domain sequence"

<400> 38

Glu Asp Asp Asp Asp Lys Ala

1 5

<210> 39

<211> 9

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

FAP cleavage domain sequence"

<400> 39

Lys Gln Glu Gln Asn Pro Gly Ser Thr

1 5

<210> 40

<211> 6

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Kallikrein cleavage domain 2" sequence

<400> 40

Gly Lys Ala Phe Arg Arg

1 5

<210> 41

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Plasmin cleavage domain sequence"

<400> 41

Asp Ala Phe Lys

1

<210> 42

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Plasmin cleavage domain sequence"

<400> 42

Asp Val Leu Lys

1

<210> 43

<211> 4

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Plasmin cleavage domain sequence"

<400> 43

Asp Ala Phe Lys

1

<210> 44

<211> 7

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

TOP cleavage domain sequence"

<400> 44

Ala Leu Leu Leu Ala Leu Leu

1 5

<210> 45

<211> 652

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 45

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp

20 25 30

Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn

85 90 95

Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val

100 105 110

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu

130 135 140

His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr

145 150 155 160

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro

165 170 175

Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu

180 185 190

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His

195 200 205

Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu

210 215 220

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr

225 230 235 240

Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser

245 250 255

Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

260 265 270

Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

275 280 285

Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe

290 295 300

Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys

305 310 315 320

Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu

325 330 335

Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

340 345 350

Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr

355 360 365

Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly

405 410 415

Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser

420 425 430

Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro

435 440 445

Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr

450 455 460

Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn

465 470 475 480

Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

485 490 495

Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp

500 505 510

Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly

515 520 525

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr

530 535 540

Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile

545 550 555 560

Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln

565 570 575

Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe

580 585 590

Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr

595 600 605

Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr

610 615 620

Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly

625 630 635 640

Thr Lys Val Glu Ile Lys His His His His His

645 650

<210> 46

<211> 652

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 46

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp

20 25 30

Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn

85 90 95

Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val

100 105 110

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

130 135 140

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser

145 150 155 160

Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

165 170 175

Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val

180 185 190

Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

195 200 205

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

210 215 220

Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr

225 230 235 240

Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

245 250 255

Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

260 265 270

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln

275 280 285

Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala

290 295 300

Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro

305 310 315 320

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

325 330 335

Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr

340 345 350

Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

355 360 365

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu

370 375 380

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser

385 390 395 400

Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly

405 410 415

Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

420 425 430

Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys

435 440 445

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu

450 455 460

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr

465 470 475 480

Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val

485 490 495

Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly

500 505 510

Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu

515 520 525

His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr

530 535 540

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro

545 550 555 560

Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu

565 570 575

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His

580 585 590

Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu

595 600 605

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr

610 615 620

Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser

625 630 635 640

Ile Ile Ser Thr Leu Thr His His His His His

645 650

<210> 47

<211> 553

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 47

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

130 135 140

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn

145 150 155 160

Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

165 170 175

Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser

180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

195 200 205

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr

210 215 220

Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly

225 230 235 240

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

245 250 255

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly

260 265 270

Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser

275 280 285

Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp

290 295 300

Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu

305 310 315 320

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu

325 330 335

Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu

340 345 350

Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp

355 360 365

Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu

370 375 380

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu

385 390 395 400

Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu

405 410 415

Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser

420 425 430

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

435 440 445

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

450 455 460

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

465 470 475 480

Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val

485 490 495

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr

500 505 510

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

515 520 525

Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr

530 535 540

Val Ser Ser His His His His His

545 550

<210> 48

<211> 553

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 48

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

130 135 140

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn

145 150 155 160

Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

165 170 175

Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser

180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

195 200 205

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr

210 215 220

Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly

225 230 235 240

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

245 250 255

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln

260 265 270

Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg

275 280 285

Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser

290 295 300

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile

305 310 315 320

Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg

325 330 335

Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met

340 345 350

Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly

355 360 365

Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser

370 375 380

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser

385 390 395 400

Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro

405 410 415

Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu

420 425 430

Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro

435 440 445

Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala

450 455 460

Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu

465 470 475 480

Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro

485 490 495

Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly

500 505 510

Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile

515 520 525

Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser

530 535 540

Thr Leu Thr His His His His His

545 550

<210> 49

<211> 553

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 49

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

1 5 10 15

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

65 70 75 80

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

100 105 110

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

115 120 125

Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

145 150 155 160

Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

165 170 175

Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly

180 185 190

Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr

195 200 205

Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys

210 215 220

Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala

225 230 235 240

Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly

245 250 255

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

260 265 270

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

275 280 285

Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

290 295 300

Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

305 310 315 320

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

325 330 335

Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly

340 345 350

Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro

355 360 365

Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn

370 375 380

Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val

385 390 395 400

Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly

405 410 415

Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

420 425 430

Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro

435 440 445

Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys

450 455 460

Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro

465 470 475 480

Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser

485 490 495

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

500 505 510

Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys

515 520 525

Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val

530 535 540

Glu Ile Lys His His His His His

545 550

<210> 50

<211> 682

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 50

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp

20 25 30

Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn

85 90 95

Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val

100 105 110

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu

130 135 140

His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr

145 150 155 160

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro

165 170 175

Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu

180 185 190

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His

195 200 205

Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu

210 215 220

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr

225 230 235 240

Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser

245 250 255

Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

260 265 270

Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

275 280 285

Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe

290 295 300

Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys

305 310 315 320

Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu

325 330 335

Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

340 345 350

Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr

355 360 365

Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

405 410 415

Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

420 425 430

Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

435 440 445

Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe

450 455 460

Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys

465 470 475 480

Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser

485 490 495

Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys

500 505 510

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

515 520 525

Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp

530 535 540

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

545 550 555 560

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser

565 570 575

Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys

580 585 590

Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys

595 600 605

Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr

610 615 620

Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe

625 630 635 640

Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr

645 650 655

Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys

660 665 670

Val Glu Ile Lys His His His His His

675 680

<210> 51

<211> 667

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 51

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp

20 25 30

Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn

85 90 95

Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val

100 105 110

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu

130 135 140

His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr

145 150 155 160

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro

165 170 175

Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu

180 185 190

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His

195 200 205

Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu

210 215 220

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr

225 230 235 240

Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser

245 250 255

Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

260 265 270

Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

275 280 285

Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe

290 295 300

Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys

305 310 315 320

Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu

325 330 335

Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

340 345 350

Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr

355 360 365

Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

405 410 415

Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly

420 425 430

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

435 440 445

Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly

450 455 460

Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr

465 470 475 480

Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

485 490 495

Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr

500 505 510

Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr

515 520 525

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ser Gly Gly Pro Gly

530 535 540

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ile Gln Met Thr Gln

545 550 555 560

Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr

565 570 575

Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln

580 585 590

Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg

595 600 605

Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

610 615 620

Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr

625 630 635 640

Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr

645 650 655

Lys Val Glu Ile Lys His His His His His

660 665

<210> 52

<211> 682

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 52

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

1 5 10 15

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

65 70 75 80

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

100 105 110

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

115 120 125

Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

145 150 155 160

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

165 170 175

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

180 185 190

Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

195 200 205

Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

210 215 220

Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr

225 230 235 240

Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu

245 250 255

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

260 265 270

Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly

275 280 285

Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

305 310 315 320

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser

325 330 335

Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys

340 345 350

Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val

355 360 365

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

370 375 380

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

385 390 395 400

Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

405 410 415

Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

420 425 430

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

435 440 445

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

450 455 460

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

465 470 475 480

Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val

485 490 495

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr

500 505 510

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

515 520 525

Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr

530 535 540

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

545 550 555 560

Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala

565 570 575

Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser

580 585 590

Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu

595 600 605

Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser

610 615 620

Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val

625 630 635 640

Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr

645 650 655

Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val

660 665 670

Thr Val Ser Ser His His His His His

675 680

<210> 53

<211> 393

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 53

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

1 5 10 15

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

65 70 75 80

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

100 105 110

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

115 120 125

Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

145 150 155 160

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

165 170 175

Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly

180 185 190

Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro

195 200 205

Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn

210 215 220

Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val

225 230 235 240

Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly

245 250 255

Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

260 265 270

Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro

275 280 285

Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys

290 295 300

Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro

305 310 315 320

Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser

325 330 335

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

340 345 350

Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys

355 360 365

Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val

370 375 380

Glu Ile Lys His His His His His

385 390

<210> 54

<211> 423

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 54

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

1 5 10 15

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

65 70 75 80

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

100 105 110

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

115 120 125

Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

145 150 155 160

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

165 170 175

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

180 185 190

Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

195 200 205

Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

210 215 220

Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr

225 230 235 240

Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu

245 250 255

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

260 265 270

Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly

275 280 285

Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

305 310 315 320

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser

325 330 335

Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys

340 345 350

Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val

355 360 365

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

370 375 380

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

385 390 395 400

Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

405 410 415

Lys His His His His His

420

<210> 55

<211> 682

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 55

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

1 5 10 15

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

65 70 75 80

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

100 105 110

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

115 120 125

Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

145 150 155 160

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

165 170 175

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

180 185 190

Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

195 200 205

Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

210 215 220

Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr

225 230 235 240

Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu

245 250 255

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

260 265 270

Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly

275 280 285

Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

305 310 315 320

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser

325 330 335

Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys

340 345 350

Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val

355 360 365

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

370 375 380

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

385 390 395 400

Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile

405 410 415

Lys Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser

420 425 430

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

435 440 445

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

450 455 460

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

465 470 475 480

Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val

485 490 495

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr

500 505 510

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

515 520 525

Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr

530 535 540

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

545 550 555 560

Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala

565 570 575

Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser

580 585 590

Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu

595 600 605

Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser

610 615 620

Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val

625 630 635 640

Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr

645 650 655

Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val

660 665 670

Thr Val Ser Ser His His His His His

675 680

<210> 56

<211> 682

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 56

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Ala Gln Ala Gly Gly

1 5 10 15

Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Asn Ser

20 25 30

Val Met Ala Trp Tyr Arg Gln Thr Pro Gly Lys Gln Arg Glu Phe Val

35 40 45

Ala Ile Ile Asn Ser Val Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Val Cys Asn

85 90 95

Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln Gly Thr Gln Val Thr Val

100 105 110

Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly

115 120 125

Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

130 135 140

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser

145 150 155 160

Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

165 170 175

Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val

180 185 190

Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

195 200 205

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

210 215 220

Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr

225 230 235 240

Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

245 250 255

Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu

260 265 270

Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln

275 280 285

Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala

290 295 300

Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro

305 310 315 320

Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

325 330 335

Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr

340 345 350

Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

355 360 365

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

370 375 380

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly

385 390 395 400

Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln

405 410 415

Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg

420 425 430

Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser

435 440 445

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile

450 455 460

Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg

465 470 475 480

Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met

485 490 495

Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly

500 505 510

Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser

515 520 525

Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala

530 535 540

Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu

545 550 555 560

Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn

565 570 575

Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys

580 585 590

Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro

595 600 605

Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg

610 615 620

Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys

625 630 635 640

Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr

645 650 655

Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile

660 665 670

Ser Thr Leu Thr His His His His His

675 680

<210> 57

<211> 393

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 57

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

130 135 140

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn

145 150 155 160

Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

165 170 175

Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser

180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

195 200 205

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr

210 215 220

Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser

225 230 235 240

Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro

245 250 255

Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu

260 265 270

Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro

275 280 285

Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala

290 295 300

Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu

305 310 315 320

Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro

325 330 335

Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly

340 345 350

Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile

355 360 365

Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser

370 375 380

Thr Leu Thr His His His His His

385 390

<210> 58

<211> 423

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 58

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Thr Leu Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

130 135 140

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn

145 150 155 160

Val Gly Thr Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

165 170 175

Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser

180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

195 200 205

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr

210 215 220

Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly

225 230 235 240

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

245 250 255

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly

260 265 270

Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser

275 280 285

Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp

290 295 300

Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu

305 310 315 320

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu

325 330 335

Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu

340 345 350

Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp

355 360 365

Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu

370 375 380

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu

385 390 395 400

Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu

405 410 415

Thr His His His His His

420

<210> 59

<211> 669

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 59

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp

20 25 30

Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn

85 90 95

Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val

100 105 110

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu

130 135 140

His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr

145 150 155 160

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro

165 170 175

Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu

180 185 190

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His

195 200 205

Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu

210 215 220

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr

225 230 235 240

Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser

245 250 255

Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

260 265 270

Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

275 280 285

Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe

290 295 300

Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys

305 310 315 320

Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu

325 330 335

Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

340 345 350

Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr

355 360 365

Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu

405 410 415

Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys Val

420 425 430

Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp

435 440 445

Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Asn

450 455 460

Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala

465 470 475 480

Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser

485 490 495

Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg

500 505 510

Gly Asp Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr

515 520 525

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

530 535 540

Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

545 550 555 560

Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser

565 570 575

Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro

580 585 590

Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser

595 600 605

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

610 615 620

Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys

625 630 635 640

Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu

645 650 655

Glu Ile Lys His His His His His Glu Pro Glu Ala

660 665

<210> 60

<211> 669

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 60

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp

20 25 30

Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn

85 90 95

Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val

100 105 110

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu

130 135 140

His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr

145 150 155 160

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro

165 170 175

Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu

180 185 190

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His

195 200 205

Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu

210 215 220

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr

225 230 235 240

Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser

245 250 255

Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

260 265 270

Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

275 280 285

Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe

290 295 300

Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys

305 310 315 320

Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu

325 330 335

Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

340 345 350

Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr

355 360 365

Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val Leu

405 410 415

Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr

420 425 430

Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr

435 440 445

Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile

450 455 460

Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly

465 470 475 480

Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Glu

485 490 495

Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp Pro Tyr

500 505 510

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser

515 520 525

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln

530 535 540

Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser Cys

545 550 555 560

Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys

565 570 575

Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro Gly

580 585 590

Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu

595 600 605

Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu

610 615 620

Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp

625 630 635 640

Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr

645 650 655

Val Ser Ser His His His His His Glu Pro Glu Ala

660 665

<210> 61

<211> 689

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 61

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

1 5 10 15

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

65 70 75 80

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

100 105 110

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

115 120 125

Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

145 150 155 160

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln

165 170 175

Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys

180 185 190

Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu

195 200 205

Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile

210 215 220

Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys

225 230 235 240

Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu

245 250 255

Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp

260 265 270

Arg Gly Asp Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr

275 280 285

Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

290 295 300

Gly Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu

305 310 315 320

Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln

325 330 335

Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys

340 345 350

Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu

355 360 365

Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe

370 375 380

Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr

385 390 395 400

Cys Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr Lys

405 410 415

Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

420 425 430

Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln

435 440 445

Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe

450 455 460

Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

465 470 475 480

Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala

485 490 495

Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr

500 505 510

Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val

515 520 525

Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr

530 535 540

Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

545 550 555 560

Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu

565 570 575

Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg

580 585 590

Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys

595 600 605

Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr

610 615 620

Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys

625 630 635 640

Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly

645 650 655

Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly

660 665 670

Thr Gln Val Thr Val Ser Ser His His His His His Glu Pro Glu

675 680 685

Ala

<210> 62

<211> 689

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 62

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

1 5 10 15

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

65 70 75 80

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

100 105 110

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

115 120 125

Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

145 150 155 160

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val

165 170 175

Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala

180 185 190

Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser

195 200 205

Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

210 215 220

Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe Ser

225 230 235 240

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu

245 250 255

Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp Pro

260 265 270

Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly

275 280 285

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln

290 295 300

Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser

305 310 315 320

Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val

325 330 335

Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro

340 345 350

Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr

355 360 365

Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser

370 375 380

Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly

385 390 395 400

Asp Gly Tyr Tyr Ala Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val

405 410 415

Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

420 425 430

Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln

435 440 445

Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe

450 455 460

Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

465 470 475 480

Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala

485 490 495

Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr

500 505 510

Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val

515 520 525

Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr

530 535 540

Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

545 550 555 560

Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu

565 570 575

Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg

580 585 590

Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys

595 600 605

Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser Tyr

610 615 620

Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys

625 630 635 640

Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly

645 650 655

Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys Gly

660 665 670

Thr Gln Val Thr Val Ser Ser His His His His His Glu Pro Glu

675 680 685

Ala

<210> 63

<211> 272

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

IL2Ra sequence"

<400> 63

Met Asp Ser Tyr Leu Leu Met Trp Gly Leu Leu Thr Phe Ile Met Val

1 5 10 15

Pro Gly Cys Gln Ala Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro

20 25 30

His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn

35 40 45

Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr

50 55 60

Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys

65 70 75 80

Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro

85 90 95

Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro

100 105 110

Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro

115 120 125

Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val

130 135 140

Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His

145 150 155 160

Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys Thr Arg

165 170 175

Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu Thr Ser Gln

180 185 190

Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly Arg Pro Glu

195 200 205

Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln Ile Gln Thr

210 215 220

Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr Glu Tyr Gln

225 230 235 240

Val Ala Val Ala Gly Cys Val Phe Leu Leu Ile Ser Val Leu Leu Leu

245 250 255

Ser Gly Leu Thr Trp Gln Arg Arg Gln Arg Lys Ser Arg Arg Thr Ile

260 265 270

<210> 64

<211> 551

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

IL2Rb sequence"

<400> 64

Met Ala Ala Pro Ala Leu Ser Trp Arg Leu Pro Leu Leu Ile Leu Leu

1 5 10 15

Leu Pro Leu Ala Thr Ser Trp Ala Ser Ala Ala Val Asn Gly Thr Ser

20 25 30

Gln Phe Thr Cys Phe Tyr Asn Ser Arg Ala Asn Ile Ser Cys Val Trp

35 40 45

Ser Gln Asp Gly Ala Leu Gln Asp Thr Ser Cys Gln Val His Ala Trp

50 55 60

Pro Asp Arg Arg Arg Trp Asn Gln Thr Cys Glu Leu Leu Pro Val Ser

65 70 75 80

Gln Ala Ser Trp Ala Cys Asn Leu Ile Leu Gly Ala Pro Asp Ser Gln

85 90 95

Lys Leu Thr Thr Val Asp Ile Val Thr Leu Arg Val Leu Cys Arg Glu

100 105 110

Gly Val Arg Trp Arg Val Met Ala Ile Gln Asp Phe Lys Pro Phe Glu

115 120 125

Asn Leu Arg Leu Met Ala Pro Ile Ser Leu Gln Val Val His Val Glu

130 135 140

Thr His Arg Cys Asn Ile Ser Trp Glu Ile Ser Gln Ala Ser His Tyr

145 150 155 160

Phe Glu Arg His Leu Glu Phe Glu Ala Arg Thr Leu Ser Pro Gly His

165 170 175

Thr Trp Glu Glu Ala Pro Leu Leu Thr Leu Lys Gln Lys Gln Glu Trp

180 185 190

Ile Cys Leu Glu Thr Leu Thr Pro Asp Thr Gln Tyr Glu Phe Gln Val

195 200 205

Arg Val Lys Pro Leu Gln Gly Glu Phe Thr Thr Trp Ser Pro Trp Ser

210 215 220

Gln Pro Leu Ala Phe Arg Thr Lys Pro Ala Ala Leu Gly Lys Asp Thr

225 230 235 240

Ile Pro Trp Leu Gly His Leu Leu Val Gly Leu Ser Gly Ala Phe Gly

245 250 255

Phe Ile Ile Leu Val Tyr Leu Leu Ile Asn Cys Arg Asn Thr Gly Pro

260 265 270

Trp Leu Lys Lys Val Leu Lys Cys Asn Thr Pro Asp Pro Ser Lys Phe

275 280 285

Phe Ser Gln Leu Ser Ser Glu His Gly Gly Asp Val Gln Lys Trp Leu

290 295 300

Ser Ser Pro Phe Pro Ser Ser Ser Phe Ser Pro Gly Gly Leu Ala Pro

305 310 315 320

Glu Ile Ser Pro Leu Glu Val Leu Glu Arg Asp Lys Val Thr Gln Leu

325 330 335

Leu Leu Gln Gln Asp Lys Val Pro Glu Pro Ala Ser Leu Ser Ser Asn

340 345 350

His Ser Leu Thr Ser Cys Phe Thr Asn Gln Gly Tyr Phe Phe Phe His

355 360 365

Leu Pro Asp Ala Leu Glu Ile Glu Ala Cys Gln Val Tyr Phe Thr Tyr

370 375 380

Asp Pro Tyr Ser Glu Glu Asp Pro Asp Glu Gly Val Ala Gly Ala Pro

385 390 395 400

Thr Gly Ser Ser Pro Gln Pro Leu Gln Pro Leu Ser Gly Glu Asp Asp

405 410 415

Ala Tyr Cys Thr Phe Pro Ser Arg Asp Asp Leu Leu Leu Phe Ser Pro

420 425 430

Ser Leu Leu Gly Gly Pro Ser Pro Pro Ser Thr Ala Pro Gly Gly Ser

435 440 445

Gly Ala Gly Glu Glu Arg Met Pro Pro Ser Leu Gln Glu Arg Val Pro

450 455 460

Arg Asp Trp Asp Pro Gln Pro Leu Gly Pro Pro Thr Pro Gly Val Pro

465 470 475 480

Asp Leu Val Asp Phe Gln Pro Pro Pro Glu Leu Val Leu Arg Glu Ala

485 490 495

Gly Glu Glu Val Pro Asp Ala Gly Pro Arg Glu Gly Val Ser Phe Pro

500 505 510

Trp Ser Arg Pro Pro Gly Gln Gly Glu Phe Arg Ala Leu Asn Ala Arg

515 520 525

Leu Pro Leu Asn Thr Asp Ala Tyr Leu Ser Leu Gln Glu Leu Gln Gly

530 535 540

Gln Asp Pro Thr His Leu Val

545 550

<210> 65

<211> 369

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

IL2Rg sequence"

<400> 65

Met Leu Lys Pro Ser Leu Pro Phe Thr Ser Leu Leu Phe Leu Gln Leu

1 5 10 15

Pro Leu Leu Gly Val Gly Leu Asn Thr Thr Ile Leu Thr Pro Asn Gly

20 25 30

Asn Glu Asp Thr Thr Ala Asp Phe Phe Leu Thr Thr Met Pro Thr Asp

35 40 45

Ser Leu Ser Val Ser Thr Leu Pro Leu Pro Glu Val Gln Cys Phe Val

50 55 60

Phe Asn Val Glu Tyr Met Asn Cys Thr Trp Asn Ser Ser Ser Glu Pro

65 70 75 80

Gln Pro Thr Asn Leu Thr Leu His Tyr Trp Tyr Lys Asn Ser Asp Asn

85 90 95

Asp Lys Val Gln Lys Cys Ser His Tyr Leu Phe Ser Glu Glu Ile Thr

100 105 110

Ser Gly Cys Gln Leu Gln Lys Lys Glu Ile His Leu Tyr Gln Thr Phe

115 120 125

Val Val Gln Leu Gln Asp Pro Arg Glu Pro Arg Arg Gln Ala Thr Gln

130 135 140

Met Leu Lys Leu Gln Asn Leu Val Ile Pro Trp Ala Pro Glu Asn Leu

145 150 155 160

Thr Leu His Lys Leu Ser Glu Ser Gln Leu Glu Leu Asn Trp Asn Asn

165 170 175

Arg Phe Leu Asn His Cys Leu Glu His Leu Val Gln Tyr Arg Thr Asp

180 185 190

Trp Asp His Ser Trp Thr Glu Gln Ser Val Asp Tyr Arg His Lys Phe

195 200 205

Ser Leu Pro Ser Val Asp Gly Gln Lys Arg Tyr Thr Phe Arg Val Arg

210 215 220

Ser Arg Phe Asn Pro Leu Cys Gly Ser Ala Gln His Trp Ser Glu Trp

225 230 235 240

Ser His Pro Ile His Trp Gly Ser Asn Thr Ser Lys Glu Asn Pro Phe

245 250 255

Leu Phe Ala Leu Glu Ala Val Val Ile Ser Val Gly Ser Met Gly Leu

260 265 270

Ile Ile Ser Leu Leu Cys Val Tyr Phe Trp Leu Glu Arg Thr Met Pro

275 280 285

Arg Ile Pro Thr Leu Lys Asn Leu Glu Asp Leu Val Thr Glu Tyr His

290 295 300

Gly Asn Phe Ser Ala Trp Ser Gly Val Ser Lys Gly Leu Ala Glu Ser

305 310 315 320

Leu Gln Pro Asp Tyr Ser Glu Arg Leu Cys Leu Val Ser Glu Ile Pro

325 330 335

Pro Lys Gly Gly Ala Leu Gly Glu Gly Pro Gly Ala Ser Pro Cys Asn

340 345 350

Gln His Ser Pro Tyr Trp Ala Pro Pro Cys Tyr Thr Leu Lys Pro Glu

355 360 365

Th

<210> 66

<211> 276

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 66

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr

65 70 75 80

Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

85 90 95

Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

100 105 110

Thr Ala Val Tyr Tyr Cys Ala Arg Gly Val Gly Ala Phe Arg Pro Tyr

115 120 125

Arg Lys His Glu Trp Gly Gln Gly Thr Leu Val Thr Val Ser Arg Gly

130 135 140

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Ser

145 150 155 160

Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val

165 170 175

Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp

180 185 190

Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys

195 200 205

Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser

210 215 220

Gly Asn Thr Ala Ser Leu Thr Thr Thr Gly Ala Gln Ala Glu Asp Glu

225 230 235 240

Ala Asp Tyr Tyr Cys Asn Ser Ser Pro Phe Glu His Asn Leu Val Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu His His His His His

260 265 270

Glu Pro Glu Ala

275

<210> 67

<211> 674

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 67

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser

210 215 220

Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser

225 230 235 240

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

245 250 255

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp

260 265 270

Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

275 280 285

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

290 295 300

Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg

305 310 315 320

Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly

325 330 335

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser

340 345 350

Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

355 360 365

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

370 375 380

Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe

385 390 395 400

Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

420 425 430

Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala

435 440 445

Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala

450 455 460

Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg

465 470 475 480

Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

485 490 495

Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro

500 505 510

Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val

515 520 525

Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser

530 535 540

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu

545 550 555 560

Ser Gly Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys

565 570 575

Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg

580 585 590

Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly

595 600 605

Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser

610 615 620

Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys

625 630 635 640

Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp

645 650 655

Tyr Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser His His His His

660 665 670

His His

<210> 68

<211> 674

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 68

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly

130 135 140

Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly

145 150 155 160

Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser

165 170 175

Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro

180 185 190

Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp

195 200 205

Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

210 215 220

Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu

225 230 235 240

Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser

245 250 255

Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro

260 265 270

Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser

275 280 285

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

290 295 300

Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln

305 310 315 320

Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser

325 330 335

Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg

340 345 350

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

355 360 365

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala

370 375 380

Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly

385 390 395 400

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln

405 410 415

Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val

420 425 430

Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp

435 440 445

Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala

450 455 460

Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser

465 470 475 480

Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe

485 490 495

Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly

500 505 510

Gly Gly Thr Lys Val Glu Ile Lys Ser Gly Gly Pro Gly Pro Ala Gly

515 520 525

Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

530 535 540

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

545 550 555 560

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

565 570 575

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

580 585 590

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

595 600 605

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

610 615 620

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

625 630 635 640

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

645 650 655

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His

660 665 670

His His

<210> 69

<211> 425

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 69

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile

20 25 30

Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu

35 40 45

Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu

50 55 60

Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln

65 70 75 80

Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr

85 90 95

Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser

100 105 110

Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu

115 120 125

Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val

130 135 140

Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu

145 150 155 160

His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys Thr

165 170 175

Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu Thr Ser

180 185 190

Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly Arg Pro

195 200 205

Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln Ile Gln

210 215 220

Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr Glu Tyr

225 230 235 240

Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

245 250 255

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser

260 265 270

Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro

275 280 285

Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu

290 295 300

Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro

305 310 315 320

Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala

325 330 335

Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu

340 345 350

Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro

355 360 365

Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly

370 375 380

Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile

385 390 395 400

Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser

405 410 415

Thr Leu Thr His His His His His

420 425

<210> 70

<211> 425

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 70

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly

165 170 175

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

180 185 190

Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro

195 200 205

Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr

210 215 220

Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly

225 230 235 240

Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp

245 250 255

Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln

260 265 270

Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met

275 280 285

Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys

290 295 300

Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His

305 310 315 320

Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg

325 330 335

Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly

340 345 350

Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu

355 360 365

Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly

370 375 380

Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln

385 390 395 400

Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr

405 410 415

Glu Tyr Gln His His His His His

420 425

<210> 71

<211> 555

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 71

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

65 70 75 80

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

85 90 95

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

100 105 110

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

115 120 125

Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

130 135 140

Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro

145 150 155 160

Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr

165 170 175

Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly

180 185 190

Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp

195 200 205

Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln

210 215 220

Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met

225 230 235 240

Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys

245 250 255

Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His

260 265 270

Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg

275 280 285

Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly

290 295 300

Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu

305 310 315 320

Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly

325 330 335

Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln

340 345 350

Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr

355 360 365

Glu Tyr Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

370 375 380

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

385 390 395 400

Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser

405 410 415

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His

420 425 430

Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys

435 440 445

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys

450 455 460

Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys

465 470 475 480

Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu

485 490 495

Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu

500 505 510

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala

515 520 525

Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile

530 535 540

Ile Ser Thr Leu Thr His His His His His

545 550 555

<210> 72

<211> 555

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 72

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly

165 170 175

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

180 185 190

Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro

195 200 205

Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr

210 215 220

Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly

225 230 235 240

Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp

245 250 255

Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln

260 265 270

Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met

275 280 285

Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys

290 295 300

Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His

305 310 315 320

Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg

325 330 335

Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly

340 345 350

Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu

355 360 365

Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly

370 375 380

Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln

385 390 395 400

Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr

405 410 415

Glu Tyr Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

420 425 430

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

435 440 445

Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

450 455 460

Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

465 470 475 480

Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu

485 490 495

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr

500 505 510

Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr

515 520 525

Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu

530 535 540

Val Thr Val Ser Ser His His His His His

545 550 555

<210> 73

<211> 555

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 73

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly

165 170 175

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

180 185 190

Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp Pro Pro

195 200 205

Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr

210 215 220

Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly

225 230 235 240

Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp

245 250 255

Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln

260 265 270

Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met

275 280 285

Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys

290 295 300

Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His

305 310 315 320

Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg

325 330 335

Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly

340 345 350

Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu

355 360 365

Thr Ser Gln Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly

370 375 380

Arg Pro Glu Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln

385 390 395 400

Ile Gln Thr Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr

405 410 415

Glu Tyr Gln Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro

420 425 430

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

435 440 445

Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

450 455 460

Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

465 470 475 480

Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu

485 490 495

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr

500 505 510

Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr

515 520 525

Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu

530 535 540

Val Thr Val Ser Ser His His His His His

545 550 555

<210> 74

<211> 575

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 74

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

65 70 75 80

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

85 90 95

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

100 105 110

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

115 120 125

Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala

130 135 140

Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly

145 150 155 160

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

165 170 175

Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala

180 185 190

Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr

195 200 205

Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp

210 215 220

Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu

225 230 235 240

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu

245 250 255

Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly

260 265 270

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met

275 280 285

Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr

290 295 300

Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr

305 310 315 320

Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser

325 330 335

Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

340 345 350

Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala

355 360 365

Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly

370 375 380

Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly

385 390 395 400

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

405 410 415

Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

420 425 430

Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu

435 440 445

Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile

450 455 460

Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe

465 470 475 480

Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu

485 490 495

Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys

500 505 510

Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile

515 520 525

Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala

530 535 540

Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe

545 550 555 560

Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His His

565 570 575

<210> 75

<211> 704

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 75

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly

165 170 175

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

180 185 190

Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

195 200 205

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

210 215 220

Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala

225 230 235 240

Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr

245 250 255

Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp

260 265 270

Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu

275 280 285

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu

290 295 300

Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ser Gly Gly

305 310 315 320

Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ile Gln Met

325 330 335

Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr

340 345 350

Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr

355 360 365

Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser

370 375 380

Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

385 390 395 400

Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala

405 410 415

Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly

420 425 430

Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly

435 440 445

Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly

450 455 460

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

465 470 475 480

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

485 490 495

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

500 505 510

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

515 520 525

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

530 535 540

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

545 550 555 560

Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly

580 585 590

Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

595 600 605

Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala

610 615 620

Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr

625 630 635 640

Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

645 650 655

Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu

660 665 670

Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp

675 680 685

Gly Lys Gly Thr Gln Val Thr Val Ser Ser His His His His His

690 695 700

<210> 76

<211> 689

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 76

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

420 425 430

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu

435 440 445

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu

450 455 460

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp

465 470 475 480

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp

485 490 495

Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr

500 505 510

Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser

515 520 525

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn

530 535 540

Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser

545 550 555 560

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

565 570 575

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

580 585 590

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn

595 600 605

Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile

610 615 620

Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

625 630 635 640

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

645 650 655

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr

660 665 670

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His

675 680 685

His

<210> 77

<211> 545

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 77

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser

210 215 220

Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser

225 230 235 240

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

245 250 255

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp

260 265 270

Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

275 280 285

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

290 295 300

Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg

305 310 315 320

Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly

325 330 335

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser

340 345 350

Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

355 360 365

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

370 375 380

Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe

385 390 395 400

Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

420 425 430

Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala

435 440 445

Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala

450 455 460

Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg

465 470 475 480

Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

485 490 495

Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro

500 505 510

Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val

515 520 525

Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His His His

530 535 540

His

545

<210> 78

<211> 575

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 78

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly

165 170 175

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

180 185 190

Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

195 200 205

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

210 215 220

Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala

225 230 235 240

Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr

245 250 255

Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp

260 265 270

Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu

275 280 285

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu

290 295 300

Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly

305 310 315 320

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met

325 330 335

Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr

340 345 350

Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr

355 360 365

Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser

370 375 380

Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

385 390 395 400

Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala

405 410 415

Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly

420 425 430

Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly

435 440 445

Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly

450 455 460

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

465 470 475 480

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

485 490 495

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

500 505 510

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

515 520 525

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

530 535 540

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

545 550 555 560

Gln Gly Thr Leu Val Thr Val Ser Ser His His His His His

565 570 575

<210> 79

<211> 421

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 79

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

65 70 75 80

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

85 90 95

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

100 105 110

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

115 120 125

Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala

130 135 140

Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys

145 150 155 160

Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile

165 170 175

Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu

180 185 190

Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu

195 200 205

Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu

210 215 220

Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn

225 230 235 240

Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met

245 250 255

Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg

260 265 270

Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly

275 280 285

Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu

290 295 300

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu

305 310 315 320

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp

325 330 335

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser

340 345 350

Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe

355 360 365

Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn

370 375 380

Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly

385 390 395 400

Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His

405 410 415

His His His His

420

<210> 80

<211> 806

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 80

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser

130 135 140

Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val Glu Ser Gly

145 150 155 160

Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala

165 170 175

Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala

180 185 190

Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg

195 200 205

Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

210 215 220

Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro

225 230 235 240

Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val

245 250 255

Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Gly Ser Gly

260 265 270

Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gln Val Gln Leu Gln

275 280 285

Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser

290 295 300

Cys Ala Ala Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr

305 310 315 320

Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg

325 330 335

Gly Gly Thr Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile

340 345 350

Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu

355 360 365

Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr

370 375 380

Asp Tyr Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly

385 390 395 400

Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln

405 410 415

Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg

420 425 430

Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala

435 440 445

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile

450 455 460

Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe

465 470 475 480

Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn

485 490 495

Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser

500 505 510

Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val

515 520 525

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

530 535 540

Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val

545 550 555 560

Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr

565 570 575

Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu

580 585 590

Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser

595 600 605

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln

610 615 620

Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro

625 630 635 640

Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser Gly Gly Pro

645 650 655

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser

660 665 670

Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu

675 680 685

Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr

690 695 700

Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu

705 710 715 720

Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val

725 730 735

Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu

740 745 750

Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr

755 760 765

Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe

770 775 780

Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr

785 790 795 800

His His His His His

805

<210> 81

<211> 676

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 81

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser

130 135 140

Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val Glu Ser Gly

145 150 155 160

Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala

165 170 175

Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala

180 185 190

Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg

195 200 205

Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

210 215 220

Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro

225 230 235 240

Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val

245 250 255

Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Ser Gly Ser Gly

260 265 270

Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val

275 280 285

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser

290 295 300

Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val

305 310 315 320

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser

325 330 335

Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile

340 345 350

Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu

355 360 365

Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp

370 375 380

Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

385 390 395 400

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp

405 410 415

Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp

420 425 430

Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val

435 440 445

Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr

450 455 460

Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser

465 470 475 480

Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu

485 490 495

Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr

500 505 510

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser Gly Gly Pro Gly Pro

515 520 525

Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr

530 535 540

Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met

545 550 555 560

Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met

565 570 575

Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His

580 585 590

Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn

595 600 605

Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser

610 615 620

Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe

625 630 635 640

Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn

645 650 655

Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His

660 665 670

His His His His

675

<210> 82

<211> 421

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 82

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser

130 135 140

Gly Ser Gly Ser Gly Ser Gly Ser Glu Val Gln Leu Val Glu Ser Gly

145 150 155 160

Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala

165 170 175

Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala

180 185 190

Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg

195 200 205

Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

210 215 220

Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro

225 230 235 240

Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val

245 250 255

Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly

260 265 270

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser

275 280 285

Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln

290 295 300

Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg

305 310 315 320

Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys

325 330 335

His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu

340 345 350

Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile

355 360 365

Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr

370 375 380

Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu

385 390 395 400

Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His

405 410 415

His His His His

420

<210> 83

<211> 420

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 83

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly

130 135 140

Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His

405 410 415

His His His His

420

<210> 84

<211> 550

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 84

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly

145 150 155 160

Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser

165 170 175

Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro

180 185 190

Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp

195 200 205

Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

210 215 220

Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu

225 230 235 240

Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser

245 250 255

Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro

260 265 270

Ala Gly Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr

275 280 285

Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met

290 295 300

Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met

305 310 315 320

Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His

325 330 335

Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn

340 345 350

Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser

355 360 365

Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe

370 375 380

Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn

385 390 395 400

Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly

405 410 415

Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln

420 425 430

Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg

435 440 445

Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser

450 455 460

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile

465 470 475 480

Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg

485 490 495

Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met

500 505 510

Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly

515 520 525

Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser

530 535 540

His His His His His

545 550

<210> 85

<211> 420

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 85

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His

405 410 415

His His His His

420

<210> 86

<211> 290

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 86

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His

275 280 285

His His

290

<210> 87

<211> 291

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 87

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser

210 215 220

Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile

225 230 235 240

Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu

245 250 255

Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu

260 265 270

Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His His

275 280 285

His His His

290

<210> 88

<211> 689

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 88

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly

130 135 140

Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

420 425 430

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu

435 440 445

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu

450 455 460

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp

465 470 475 480

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp

485 490 495

Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr

500 505 510

Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser

515 520 525

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn

530 535 540

Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser

545 550 555 560

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

565 570 575

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

580 585 590

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn

595 600 605

Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile

610 615 620

Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

625 630 635 640

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

645 650 655

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr

660 665 670

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His

675 680 685

His

<210> 89

<211> 704

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 89

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Gly Gly Gly Gly Ser Gly

165 170 175

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

180 185 190

Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

195 200 205

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

210 215 220

Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala

225 230 235 240

Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr

245 250 255

Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp

260 265 270

Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu

275 280 285

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu

290 295 300

Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly

305 310 315 320

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met

325 330 335

Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr

340 345 350

Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr

355 360 365

Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser

370 375 380

Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

385 390 395 400

Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala

405 410 415

Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly

420 425 430

Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly

435 440 445

Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly

450 455 460

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

465 470 475 480

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

485 490 495

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

500 505 510

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

515 520 525

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

530 535 540

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

545 550 555 560

Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly

580 585 590

Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala

595 600 605

Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr

610 615 620

Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser

625 630 635 640

Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

645 650 655

Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu

660 665 670

Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp

675 680 685

Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His His

690 695 700

<210> 90

<211> 674

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 90

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly

50 55 60

Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn

65 70 75 80

Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr

100 105 110

Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Ala Pro Thr Ser Ser Ser Thr Lys

130 135 140

Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile

145 150 155 160

Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu

165 170 175

Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu

180 185 190

Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu

195 200 205

Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn

210 215 220

Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met

225 230 235 240

Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg

245 250 255

Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly

260 265 270

Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu

275 280 285

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu

290 295 300

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp

305 310 315 320

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser

325 330 335

Gly Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe

340 345 350

Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn

355 360 365

Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly

370 375 380

Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly

385 390 395 400

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln

420 425 430

Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg

435 440 445

Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala

450 455 460

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile

465 470 475 480

Asp Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe

485 490 495

Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn

500 505 510

Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser

515 520 525

Asn Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val

530 535 540

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

545 550 555 560

Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val

565 570 575

Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr

580 585 590

Asn Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu

595 600 605

Ile Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser

610 615 620

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln

625 630 635 640

Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro

645 650 655

Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His

660 665 670

His His

<210> 91

<211> 704

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 91

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr

65 70 75 80

Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr

100 105 110

Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr

115 120 125

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser

130 135 140

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln

145 150 155 160

Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr

165 170 175

Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln

180 185 190

Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg

195 200 205

Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

210 215 220

Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr

225 230 235 240

Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr

245 250 255

Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

260 265 270

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

275 280 285

Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro

290 295 300

Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu

305 310 315 320

Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn

325 330 335

Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met

340 345 350

Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu

355 360 365

Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe

370 375 380

His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu

385 390 395 400

Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu

405 410 415

Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln

420 425 430

Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly Pro Ala Gly Met

435 440 445

Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly

450 455 460

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

465 470 475 480

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

485 490 495

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

500 505 510

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

515 520 525

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

530 535 540

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

545 550 555 560

Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly

580 585 590

Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala

595 600 605

Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr

610 615 620

Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser

625 630 635 640

Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

645 650 655

Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu

660 665 670

Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp

675 680 685

Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His His

690 695 700

<210> 92

<211> 704

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 92

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly

50 55 60

Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn

65 70 75 80

Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr

100 105 110

Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly

145 150 155 160

Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser

165 170 175

Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro

180 185 190

Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp

195 200 205

Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

210 215 220

Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu

225 230 235 240

Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser

245 250 255

Ser Gln Gly Thr Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro

260 265 270

Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu Ser

275 280 285

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

290 295 300

Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln

305 310 315 320

Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser

325 330 335

Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg

340 345 350

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

355 360 365

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala

370 375 380

Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly

385 390 395 400

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln

405 410 415

Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val

420 425 430

Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp

435 440 445

Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala

450 455 460

Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser

465 470 475 480

Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe

485 490 495

Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly

500 505 510

Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly

515 520 525

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

530 535 540

Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

545 550 555 560

Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln

565 570 575

Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly

580 585 590

Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys

595 600 605

Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu

610 615 620

Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser

625 630 635 640

Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val

645 650 655

Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr

660 665 670

Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr

675 680 685

Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His His

690 695 700

<210> 93

<211> 689

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 93

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr

65 70 75 80

Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr

100 105 110

Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr

115 120 125

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser

130 135 140

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln

145 150 155 160

Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr

165 170 175

Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln

180 185 190

Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg

195 200 205

Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

210 215 220

Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr

225 230 235 240

Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr

245 250 255

Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

260 265 270

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

275 280 285

Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln

290 295 300

Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe

305 310 315 320

Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

325 330 335

Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala

340 345 350

Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr

355 360 365

Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val

370 375 380

Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr

385 390 395 400

Leu Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly Met Lys

405 410 415

Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln

420 425 430

Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly

435 440 445

Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys

450 455 460

Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu

465 470 475 480

Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser

485 490 495

Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val

500 505 510

Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr

515 520 525

Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr

530 535 540

Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser Gly

545 550 555 560

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser

565 570 575

Gly Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala

580 585 590

Ala Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln

595 600 605

Thr Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly

610 615 620

Ser Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

625 630 635 640

Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro

645 650 655

Glu Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr

660 665 670

Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His

675 680 685

His

<210> 94

<211> 700

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 94

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Arg Gly Glu Thr Gly

290 295 300

Pro Ala Ala Pro Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly

305 310 315 320

Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

325 330 335

Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly

340 345 350

Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr

355 360 365

Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

370 375 380

Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp

385 390 395 400

Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser Val Ser Ser

405 410 415

Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

420 425 430

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

435 440 445

Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly

450 455 460

Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser

465 470 475 480

Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro

485 490 495

Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr

500 505 510

Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn

515 520 525

Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

530 535 540

Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp

545 550 555 560

Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly

565 570 575

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr

580 585 590

Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile

595 600 605

Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln

610 615 620

Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe

625 630 635 640

Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr

645 650 655

Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr

660 665 670

Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly

675 680 685

Thr Lys Val Glu Ile Lys His His His His His

690 695 700

<210> 95

<211> 276

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 95

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr

65 70 75 80

Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

85 90 95

Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

100 105 110

Thr Ala Val Tyr Tyr Cys Ala Arg Gly Val Gly Ala Phe Arg Pro Tyr

115 120 125

Arg Lys His Glu Trp Gly Gln Gly Thr Leu Val Thr Val Ser Arg Gly

130 135 140

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Ser

145 150 155 160

Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val

165 170 175

Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp

180 185 190

Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys

195 200 205

Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser

210 215 220

Gly Asn Thr Ala Ser Leu Thr Thr Thr Gly Ala Gln Ala Glu Asp Glu

225 230 235 240

Ala Asp Tyr Tyr Cys Asn Ser Ser Pro Phe Glu His Asn Leu Val Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu His His His His His

260 265 270

Glu Pro Glu Ala

275

<210> 96

<211> 279

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 96

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Gln Ser Gly Ala Glu

20 25 30

Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly

35 40 45

Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly

50 55 60

Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr

65 70 75 80

Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys

85 90 95

Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp

100 105 110

Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala

115 120 125

Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly

130 135 140

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

145 150 155 160

Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg

165 170 175

Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp

180 185 190

Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

195 200 205

Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val

225 230 235 240

Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp

245 250 255

Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys His His His

260 265 270

His His His Glu Pro Glu Ala

275

<210> 97

<211> 279

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 97

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Gln Ser Gly Ala Glu

20 25 30

Leu Val Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly

35 40 45

Tyr Ala Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly

50 55 60

Gln Gly Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr

65 70 75 80

Asn Tyr Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys

85 90 95

Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp

100 105 110

Ser Ala Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala

115 120 125

Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Ser

130 135 140

Gly Gly Pro Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ile

145 150 155 160

Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg

165 170 175

Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp

180 185 190

Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

195 200 205

Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val

225 230 235 240

Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Asn Glu Asp

245 250 255

Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys His His His

260 265 270

His His His Glu Pro Glu Ala

275

<210> 98

<211> 279

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 98

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Asp Ile Val Leu Thr Gln Ser Pro Ala Ser

20 25 30

Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser

35 40 45

Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu

65 70 75 80

Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr

100 105 110

Tyr Cys Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr

115 120 125

Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

130 135 140

Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val

145 150 155 160

Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala

165 170 175

Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly

180 185 190

Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr

195 200 205

Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser

210 215 220

Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala

225 230 235 240

Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala Tyr Phe

245 250 255

Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser His His His

260 265 270

His His His Glu Pro Glu Ala

275

<210> 99

<211> 279

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 99

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Asp Ile Val Leu Thr Gln Ser Pro Ala Ser

20 25 30

Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser

35 40 45

Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu

65 70 75 80

Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr

100 105 110

Tyr Cys Gln Gln Ser Asn Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr

115 120 125

Lys Leu Glu Ile Lys Ser Gly Gly Pro Gly Pro Ala Gly Met Lys Gly

130 135 140

Leu Pro Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val

145 150 155 160

Arg Pro Gly Thr Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala

165 170 175

Phe Thr Asn Tyr Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly

180 185 190

Leu Glu Trp Ile Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr

195 200 205

Asn Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser

210 215 220

Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala

225 230 235 240

Val Tyr Phe Cys Ala Arg Trp Arg Gly Asp Gly Tyr Tyr Ala Tyr Phe

245 250 255

Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser His His His

260 265 270

His His His Glu Pro Glu Ala

275

<210> 100

<211> 295

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 100

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser

145 150 155 160

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser

210 215 220

Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile

225 230 235 240

Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu

245 250 255

Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu

260 265 270

Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser His His His

275 280 285

His His His Glu Pro Glu Ala

290 295

<210> 101

<211> 765

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 101

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser

145 150 155 160

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ala His Lys Ser Glu

165 170 175

Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu

180 185 190

Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp

195 200 205

His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val

210 215 220

Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe

225 230 235 240

Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu

245 250 255

Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe

260 265 270

Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro

275 280 285

Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe

290 295 300

Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr

305 310 315 320

Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr

325 330 335

Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu

340 345 350

Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu

355 360 365

Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp

370 375 380

Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu

385 390 395 400

Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys

405 410 415

His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys

420 425 430

Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys

435 440 445

Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu

450 455 460

Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val

465 470 475 480

Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe

485 490 495

Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser

500 505 510

Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu

515 520 525

Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe

530 535 540

Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln

545 550 555 560

Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala

565 570 575

Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr

580 585 590

Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys

595 600 605

Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser

610 615 620

Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser

625 630 635 640

Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro

645 650 655

Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe

660 665 670

Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu

675 680 685

Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys

690 695 700

His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp

705 710 715 720

Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr

725 730 735

Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala

740 745 750

Leu Gly Leu His His His His His Glu Pro Glu Ala

755 760 765

<210> 102

<211> 765

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 102

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser

145 150 155 160

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ala His Lys Ser Glu

165 170 175

Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu

180 185 190

Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp

195 200 205

His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val

210 215 220

Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe

225 230 235 240

Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu

245 250 255

Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe

260 265 270

Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro

275 280 285

Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe

290 295 300

Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr

305 310 315 320

Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr

325 330 335

Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu

340 345 350

Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu

355 360 365

Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp

370 375 380

Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu

385 390 395 400

Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys

405 410 415

His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys

420 425 430

Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys

435 440 445

Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu

450 455 460

Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val

465 470 475 480

Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe

485 490 495

Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser

500 505 510

Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu

515 520 525

Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe

530 535 540

Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln

545 550 555 560

Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala

565 570 575

Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr

580 585 590

Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys

595 600 605

Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser

610 615 620

Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser

625 630 635 640

Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro

645 650 655

Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe

660 665 670

Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu

675 680 685

Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys

690 695 700

His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp

705 710 715 720

Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr

725 730 735

Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala

740 745 750

Leu Gly Leu His His His His His Glu Pro Glu Ala

755 760 765

<210> 103

<211> 765

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 103

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Asp Ala His Lys Ser Glu

165 170 175

Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu

180 185 190

Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp

195 200 205

His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val

210 215 220

Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe

225 230 235 240

Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu

245 250 255

Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe

260 265 270

Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro

275 280 285

Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe

290 295 300

Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr

305 310 315 320

Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr

325 330 335

Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu

340 345 350

Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu

355 360 365

Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp

370 375 380

Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu

385 390 395 400

Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys

405 410 415

His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys

420 425 430

Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys

435 440 445

Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu

450 455 460

Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val

465 470 475 480

Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe

485 490 495

Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser

500 505 510

Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu

515 520 525

Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe

530 535 540

Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln

545 550 555 560

Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala

565 570 575

Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr

580 585 590

Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys

595 600 605

Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser

610 615 620

Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser

625 630 635 640

Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro

645 650 655

Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe

660 665 670

Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu

675 680 685

Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys

690 695 700

His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp

705 710 715 720

Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr

725 730 735

Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala

740 745 750

Leu Gly Leu His His His His His Glu Pro Glu Ala

755 760 765

<210> 104

<211> 708

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 104

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser

210 215 220

Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile

225 230 235 240

Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu

245 250 255

Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu

260 265 270

Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly

275 280 285

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly

305 310 315 320

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu

325 330 335

Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys

340 345 350

Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg

355 360 365

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser

370 375 380

Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser

385 390 395 400

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

405 410 415

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp

420 425 430

Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

450 455 460

Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg

465 470 475 480

Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly

485 490 495

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser

500 505 510

Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

515 520 525

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

530 535 540

Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe

545 550 555 560

Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly

580 585 590

Gly Gly Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala

595 600 605

Ser Gly Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr

610 615 620

Pro Gly Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser

625 630 635 640

Thr Asn Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

645 650 655

Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu

660 665 670

Asp Thr Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp

675 680 685

Gly Gln Gly Thr Gln Val Thr Val Ser Ser His His His His His

690 695 700

Glu Pro Glu Ala

705

<210> 105

<211> 708

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 105

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly

50 55 60

Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn

65 70 75 80

Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr

100 105 110

Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

420 425 430

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro

435 440 445

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

450 455 460

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser

465 470 475 480

Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val

485 490 495

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser

500 505 510

Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile

515 520 525

Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu

530 535 540

Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp

545 550 555 560

Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp

580 585 590

Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp

595 600 605

Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val

610 615 620

Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr

625 630 635 640

Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser

645 650 655

Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu

660 665 670

Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr

675 680 685

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His

690 695 700

Glu Pro Glu Ala

705

<210> 106

<211> 708

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 106

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Ala Gln Ala Gly Gly Ser Leu Ser Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Val Ser Asn Ser Val Met Ala Trp Tyr Arg Gln Thr Pro Gly

50 55 60

Lys Gln Arg Glu Phe Val Ala Ile Ile Asn Ser Val Gly Ser Thr Asn

65 70 75 80

Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr

100 105 110

Ala Val Tyr Val Cys Asn Arg Asn Phe Asp Arg Ile Tyr Trp Gly Gln

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly

130 135 140

Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

420 425 430

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro

435 440 445

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

450 455 460

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser

465 470 475 480

Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val

485 490 495

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser

500 505 510

Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile

515 520 525

Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu

530 535 540

Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp

545 550 555 560

Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp

580 585 590

Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp

595 600 605

Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val

610 615 620

Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr

625 630 635 640

Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser

645 650 655

Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu

660 665 670

Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr

675 680 685

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His

690 695 700

Glu Pro Glu Ala

705

<210> 107

<211> 161

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 107

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr His His His His

145 150 155 160

His

<210> 108

<211> 708

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 108

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser

210 215 220

Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile

225 230 235 240

Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu

245 250 255

Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu

260 265 270

Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly

275 280 285

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly

305 310 315 320

Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val Glu

325 330 335

Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys

340 345 350

Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg

355 360 365

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser

370 375 380

Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser

385 390 395 400

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

405 410 415

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp

420 425 430

Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

450 455 460

Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg

465 470 475 480

Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly

485 490 495

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser

500 505 510

Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

515 520 525

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

530 535 540

Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe

545 550 555 560

Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly

580 585 590

Gly Gly Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

595 600 605

Ser Gly Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala

610 615 620

Pro Gly Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr

625 630 635 640

Ile Ser Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

645 650 655

Asn Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu

660 665 670

Asp Thr Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp

675 680 685

Gly Lys Gly Thr Gln Val Thr Val Ser Ser His His His His His

690 695 700

Glu Pro Glu Ala

705

<210> 109

<211> 708

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 109

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

130 135 140

Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

420 425 430

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro

435 440 445

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

450 455 460

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser

465 470 475 480

Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val

485 490 495

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser

500 505 510

Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile

515 520 525

Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu

530 535 540

Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp

545 550 555 560

Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp

580 585 590

Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp

595 600 605

Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val

610 615 620

Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr

625 630 635 640

Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser

645 650 655

Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu

660 665 670

Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr

675 680 685

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His

690 695 700

Glu Pro Glu Ala

705

<210> 110

<211> 708

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 110

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Ala Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Arg Ile Phe Ser Ile Asp Ile Met Ser Trp Tyr Arg Gln Ala Pro Gly

50 55 60

Lys Gln Arg Glu Leu Val Ala Arg Ile Thr Arg Gly Gly Thr Ile Ser

65 70 75 80

Tyr Asp Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr

100 105 110

Gly Val Tyr Tyr Cys Asn Ala Leu Tyr Gly Thr Asp Tyr Trp Gly Lys

115 120 125

Gly Thr Gln Val Thr Val Ser Ser Ser Gly Gly Pro Gly Pro Ala Gly

130 135 140

Met Lys Gly Leu Pro Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys

145 150 155 160

Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu

165 170 175

Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr

180 185 190

Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln

195 200 205

Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala

210 215 220

Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile

225 230 235 240

Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys

245 250 255

Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp

260 265 270

Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro

275 280 285

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

290 295 300

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

305 310 315 320

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

325 330 335

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

340 345 350

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

355 360 365

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

370 375 380

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

385 390 395 400

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

405 410 415

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

420 425 430

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro

435 440 445

Gly Pro Ala Gly Met Lys Gly Leu Pro Gly Ser Glu Val Gln Leu Val

450 455 460

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser

465 470 475 480

Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val

485 490 495

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser

500 505 510

Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile

515 520 525

Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu

530 535 540

Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp

545 550 555 560

Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

565 570 575

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp

580 585 590

Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp

595 600 605

Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val

610 615 620

Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr

625 630 635 640

Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser

645 650 655

Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu

660 665 670

Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr

675 680 685

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His

690 695 700

Glu Pro Glu Ala

705

<210> 111

<211> 579

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 111

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Gly

145 150 155 160

Pro Ala Gly Leu Tyr Ala Gln Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser

210 215 220

Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile

225 230 235 240

Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu

245 250 255

Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu

260 265 270

Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly

275 280 285

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Gly

305 310 315 320

Pro Ala Gly Leu Tyr Ala Gln Pro Gly Ser Glu Val Gln Leu Val Glu

325 330 335

Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys

340 345 350

Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg

355 360 365

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser

370 375 380

Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser

385 390 395 400

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

405 410 415

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp

420 425 430

Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

450 455 460

Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg

465 470 475 480

Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly

485 490 495

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser

500 505 510

Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

515 520 525

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

530 535 540

Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe

545 550 555 560

Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His Glu

565 570 575

Pro Glu Ala

<210> 112

<211> 579

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 112

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Pro

145 150 155 160

Gly Gly Pro Ala Gly Ile Gly Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser

210 215 220

Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile

225 230 235 240

Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu

245 250 255

Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu

260 265 270

Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly

275 280 285

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Pro

305 310 315 320

Gly Gly Pro Ala Gly Ile Gly Pro Gly Ser Glu Val Gln Leu Val Glu

325 330 335

Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys

340 345 350

Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg

355 360 365

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser

370 375 380

Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser

385 390 395 400

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

405 410 415

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp

420 425 430

Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

450 455 460

Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg

465 470 475 480

Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly

485 490 495

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser

500 505 510

Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

515 520 525

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

530 535 540

Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe

545 550 555 560

Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His Glu

565 570 575

Pro Glu Ala

<210> 113

<211> 579

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 113

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Ala

145 150 155 160

Leu Phe Lys Ser Ser Phe Pro Pro Gly Ser Glu Val Gln Leu Val Glu

165 170 175

Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser Cys

180 185 190

Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val Arg

195 200 205

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser

210 215 220

Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile

225 230 235 240

Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu

245 250 255

Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu

260 265 270

Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly

275 280 285

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro Ala

305 310 315 320

Leu Phe Lys Ser Ser Phe Pro Pro Gly Ser Glu Val Gln Leu Val Glu

325 330 335

Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys

340 345 350

Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg

355 360 365

Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser

370 375 380

Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser

385 390 395 400

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

405 410 415

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp

420 425 430

Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

450 455 460

Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg

465 470 475 480

Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly

485 490 495

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser

500 505 510

Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

515 520 525

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

530 535 540

Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe

545 550 555 560

Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His His Glu

565 570 575

Pro Glu Ala

<210> 114

<211> 581

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 114

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Pro

145 150 155 160

Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu Val

165 170 175

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

180 185 190

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

195 200 205

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

210 215 220

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

225 230 235 240

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

245 250 255

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

260 265 270

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

275 280 285

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

290 295 300

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro

305 310 315 320

Pro Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu

325 330 335

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu

340 345 350

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp

355 360 365

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp

370 375 380

Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr

385 390 395 400

Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser

405 410 415

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn

420 425 430

Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser

435 440 445

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

465 470 475 480

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn

485 490 495

Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile

500 505 510

Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

515 520 525

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

530 535 540

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr

545 550 555 560

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His

565 570 575

His Glu Pro Glu Ala

580

<210> 115

<211> 613

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 115

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr

20 25 30

Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn

35 40 45

Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe

50 55 60

Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys

65 70 75 80

Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln

85 90 95

Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn

100 105 110

Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu

115 120 125

Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile

130 135 140

Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr Ser Gly Gly Pro Pro

145 150 155 160

Gly Gly Pro Ala Gly Ile Gly Ala Leu Phe Lys Ser Ser Phe Pro Pro

165 170 175

Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu Val

180 185 190

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

195 200 205

Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp Val

210 215 220

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

225 230 235 240

Ser Gly Arg Asp Thr Leu Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr

245 250 255

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

260 265 270

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

275 280 285

Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

290 295 300

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

305 310 315 320

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Gly Gly Pro

325 330 335

Pro Gly Gly Pro Ala Gly Ile Gly Ala Leu Phe Lys Ser Ser Phe Pro

340 345 350

Pro Leu Ala Gln Lys Leu Lys Ser Ser Pro Gly Ser Glu Val Gln Leu

355 360 365

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu

370 375 380

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp

385 390 395 400

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Ala Ile Asp

405 410 415

Ser Ser Ser Tyr Thr Tyr Ser Pro Asp Thr Val Arg Gly Arg Phe Thr

420 425 430

Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser

435 440 445

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn

450 455 460

Trp Asp Ala Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser

465 470 475 480

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

485 490 495

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

500 505 510

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn

515 520 525

Val Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile

530 535 540

Tyr Ser Ala Ser Phe Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

545 550 555 560

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

565 570 575

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr

580 585 590

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys His His His His

595 600 605

His Glu Pro Glu Ala

610

<210> 116

<211> 534

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 116

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn

35 40 45

Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly

50 55 60

Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser

65 70 75 80

Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg

85 90 95

Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser

100 105 110

Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

115 120 125

Lys Pro Leu Gly Leu Gln Ala Arg Val Val Gly Gly Gly Gly Thr Gln

130 135 140

Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr

145 150 155 160

Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn

165 170 175

Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu

180 185 190

Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser

195 200 205

Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln

210 215 220

Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg

225 230 235 240

Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly

245 250 255

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val

260 265 270

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser

275 280 285

Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val

290 295 300

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser

305 310 315 320

Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg

325 330 335

Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met

340 345 350

Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His

355 360 365

Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

405 410 415

Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser

420 425 430

Tyr Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe

435 440 445

Val Val Ala Ile Asn Trp Ala Ser Gly Ser Thr Tyr Tyr Ala Asp Ser

450 455 460

Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu

465 470 475 480

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

485 490 495

Cys Ala Ala Gly Tyr Gln Ile Asn Ser Gly Asn Tyr Asn Phe Lys Asp

500 505 510

Tyr Glu Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

515 520 525

His His His His His

530

<210> 117

<211> 534

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 117

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn

35 40 45

Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly

50 55 60

Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser

65 70 75 80

Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg

85 90 95

Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser

100 105 110

Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Gly Gly Val Val Gly Gly Gly Gly Thr Gln

130 135 140

Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr

145 150 155 160

Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn

165 170 175

Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu

180 185 190

Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser

195 200 205

Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln

210 215 220

Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg

225 230 235 240

Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly

245 250 255

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val

260 265 270

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser

275 280 285

Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val

290 295 300

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser

305 310 315 320

Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg

325 330 335

Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met

340 345 350

Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His

355 360 365

Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

405 410 415

Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser

420 425 430

Tyr Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe

435 440 445

Val Val Ala Ile Asn Trp Ala Ser Gly Ser Thr Tyr Tyr Ala Asp Ser

450 455 460

Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu

465 470 475 480

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

485 490 495

Cys Ala Ala Gly Tyr Gln Ile Asn Ser Gly Asn Tyr Asn Phe Lys Asp

500 505 510

Tyr Glu Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

515 520 525

His His His His His

530

<210> 118

<211> 398

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 118

Val Val Gly Gly Gly Gly Thr Gln Thr Val Val Thr Gln Glu Pro Ser

1 5 10 15

Leu Thr Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser

20 25 30

Thr Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys

35 40 45

Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val

50 55 60

Pro Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala

65 70 75 80

Ala Leu Thr Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr

85 90 95

Cys Thr Leu Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys

100 105 110

Leu Thr Val Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

115 120 125

Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln

130 135 140

Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe

145 150 155 160

Asn Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

165 170 175

Glu Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr

180 185 190

Tyr Ala Asp Gln Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser

195 200 205

Lys Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Val Arg His Ala Asn Phe Gly Asn Ser Tyr Ile

225 230 235 240

Ser Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

245 250 255

Gly Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser

260 265 270

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Thr Leu Ser Cys Ala

275 280 285

Ala Ser Gly Arg Thr Phe Ser Ser Tyr Ala Met Gly Trp Phe Arg Gln

290 295 300

Ala Pro Gly Lys Glu Arg Glu Phe Val Val Ala Ile Asn Trp Ala Ser

305 310 315 320

Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser

325 330 335

Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg

340 345 350

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Ala Gly Tyr Gln Ile Asn

355 360 365

Ser Gly Asn Tyr Asn Phe Lys Asp Tyr Glu Tyr Asp Tyr Trp Gly Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser His His His His His

385 390 395

<210> 119

<211> 518

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 119

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn

35 40 45

Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly

50 55 60

Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser

65 70 75 80

Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg

85 90 95

Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser

100 105 110

Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

115 120 125

Lys Pro Leu Gly Leu Gln Ala Arg Val Val Gly Gly Gly Gly Thr Gln

130 135 140

Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr

145 150 155 160

Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn

165 170 175

Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu

180 185 190

Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser

195 200 205

Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln

210 215 220

Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg

225 230 235 240

Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly

245 250 255

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val

260 265 270

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser

275 280 285

Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val

290 295 300

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser

305 310 315 320

Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg

325 330 335

Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met

340 345 350

Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His

355 360 365

Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

405 410 415

Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Arg Phe Met Ile Ser Glu

420 425 430

Tyr His Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

435 440 445

Val Ser Thr Ile Asn Pro Ala Gly Thr Thr Asp Tyr Ala Glu Ser Val

450 455 460

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

465 470 475 480

Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys

485 490 495

Asp Ser Tyr Gly Tyr Arg Gly Gln Gly Thr Gln Val Thr Val Ser Ser

500 505 510

His His His His His

515

<210> 120

<211> 518

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 120

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn

35 40 45

Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly

50 55 60

Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser

65 70 75 80

Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg

85 90 95

Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser

100 105 110

Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Gly Gly Val Val Gly Gly Gly Gly Thr Gln

130 135 140

Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr

145 150 155 160

Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Gly Asn

165 170 175

Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu

180 185 190

Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr Pro Ala Arg Phe Ser

195 200 205

Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln

210 215 220

Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu Trp Tyr Ser Asn Arg

225 230 235 240

Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly

245 250 255

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val

260 265 270

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser

275 280 285

Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val

290 295 300

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser

305 310 315 320

Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg

325 330 335

Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met

340 345 350

Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His

355 360 365

Ala Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln

370 375 380

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

385 390 395 400

Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

405 410 415

Gly Ser Leu Thr Leu Ser Cys Ala Ala Ser Arg Phe Met Ile Ser Glu

420 425 430

Tyr His Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

435 440 445

Val Ser Thr Ile Asn Pro Ala Gly Thr Thr Asp Tyr Ala Glu Ser Val

450 455 460

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

465 470 475 480

Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys

485 490 495

Asp Ser Tyr Gly Tyr Arg Gly Gln Gly Thr Gln Val Thr Val Ser Ser

500 505 510

His His His His His

515

<210> 121

<211> 382

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 121

Val Val Gly Gly Gly Gly Thr Gln Thr Val Val Thr Gln Glu Pro Ser

1 5 10 15

Leu Thr Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser

20 25 30

Thr Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys

35 40 45

Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val

50 55 60

Pro Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala

65 70 75 80

Ala Leu Thr Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr

85 90 95

Cys Thr Leu Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys

100 105 110

Leu Thr Val Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

115 120 125

Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln

130 135 140

Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe

145 150 155 160

Asn Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

165 170 175

Glu Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr

180 185 190

Tyr Ala Asp Gln Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser

195 200 205

Lys Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr

210 215 220

Ala Val Tyr Tyr Cys Val Arg His Ala Asn Phe Gly Asn Ser Tyr Ile

225 230 235 240

Ser Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

245 250 255

Gly Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser

260 265 270

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Thr Leu Ser Cys Ala

275 280 285

Ala Ser Arg Phe Met Ile Ser Glu Tyr His Met His Trp Val Arg Gln

290 295 300

Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Thr Ile Asn Pro Ala Gly

305 310 315 320

Thr Thr Asp Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

325 330 335

Asp Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro

340 345 350

Glu Asp Thr Ala Val Tyr Tyr Cys Asp Ser Tyr Gly Tyr Arg Gly Gln

355 360 365

Gly Thr Gln Val Thr Val Ser Ser His His His His His

370 375 380

<210> 122

<211> 503

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 122

Gln Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr

20 25 30

Ser Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val

35 40 45

Ala Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ala Arg Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Asn Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr

100 105 110

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu

115 120 125

Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu

130 135 140

Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe Gly Met Ser Trp

145 150 155 160

Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser

165 170 175

Gly Ser Gly Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe

180 185 190

Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn

195 200 205

Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly

210 215 220

Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly

225 230 235 240

Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

245 250 255

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala

260 265 270

Ser Gly Phe Thr Phe Asn Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala

275 280 285

Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn

290 295 300

Asn Tyr Ala Thr Tyr Tyr Ala Asp Gln Val Lys Asp Arg Phe Thr Ile

305 310 315 320

Ser Arg Asp Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu

325 330 335

Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg His Ala Asn Phe

340 345 350

Gly Asn Ser Tyr Ile Ser Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu

355 360 365

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

370 375 380

Gly Gly Gly Ser Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val

385 390 395 400

Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala

405 410 415

Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln

420 425 430

Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val Pro Gly Thr

435 440 445

Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr

450 455 460

Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Thr Leu

465 470 475 480

Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val

485 490 495

Leu His His His His His

500

<210> 123

<211> 509

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 123

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ser Gly Ser Gly Arg Asp Thr Leu Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Ile Gly Gly Ser Leu Ser Val Ser Ser Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Val

115 120 125

Val Gly Gly Gly Gly Thr Gln Thr Val Val Thr Gln Glu Pro Ser Leu

130 135 140

Thr Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Ala Ser Ser Thr

145 150 155 160

Gly Ala Val Thr Ser Gly Asn Tyr Pro Asn Trp Val Gln Gln Lys Pro

165 170 175

Gly Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Lys Phe Leu Val Pro

180 185 190

Gly Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala

195 200 205

Leu Thr Leu Ser Gly Val Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys

210 215 220

Thr Leu Trp Tyr Ser Asn Arg Trp Val Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Thr Val Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

245 250 255

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

260 265 270

Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn

275 280 285

Lys Tyr Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

290 295 300

Trp Val Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr

305 310 315 320

Ala Asp Gln Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys

325 330 335

Asn Thr Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala

340 345 350

Val Tyr Tyr Cys Val Arg His Ala Asn Phe Gly Asn Ser Tyr Ile Ser

355 360 365

Tyr Trp Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly

370 375 380

Gly Gly Gly Ser Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

385 390 395 400

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Thr Leu Ser Cys Ala Ala

405 410 415

Ser Arg Phe Met Ile Ser Glu Tyr His Met His Trp Val Arg Gln Ala

420 425 430

Pro Gly Lys Gly Leu Glu Trp Val Ser Thr Ile Asn Pro Ala Gly Thr

435 440 445

Thr Asp Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

450 455 460

Asn Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu

465 470 475 480

Asp Thr Ala Val Tyr Tyr Cys Asp Ser Tyr Gly Tyr Arg Gly Gln Gly

485 490 495

Thr Gln Val Thr Val Ser Ser His His His His His

500 505

<210> 124

<211> 267

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 124

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Leu Arg Gly Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly

20 25 30

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

35 40 45

Phe Thr Phe Ser Ser Tyr Thr Leu Ala Trp Val Arg Gln Ala Pro Gly

50 55 60

Lys Gly Leu Glu Trp Val Ala Ala Ile Asp Ser Ser Ser Tyr Thr Tyr

65 70 75 80

Ser Pro Asp Thr Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

85 90 95

Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr

100 105 110

Ala Val Tyr Tyr Cys Ala Arg Asp Ser Asn Trp Asp Ala Leu Asp Tyr

115 120 125

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser

130 135 140

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln

145 150 155 160

Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr

165 170 175

Cys Lys Ala Ser Gln Asn Val Gly Thr Asn Val Gly Trp Tyr Gln Gln

180 185 190

Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile Tyr Ser Ala Ser Phe Arg

195 200 205

Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

210 215 220

Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr

225 230 235 240

Tyr Cys Gln Gln Tyr Tyr Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr

245 250 255

Lys Val Glu Ile Lys His His His His His

260 265

<210> 125

<211> 5

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

Sortase A cleavage site"

<220>

<221> MOD_OST

<222> (3)..(3)

<223> Any amino acid

<400> 125

Leu Pro Xaa Thr Gly

1 5

<210> 126

<211> 25

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

peptide"

<220>

<221> SITE

<222> (1)..(25)

<223> /note="This sequence may contain 1-5 repeats

links 'Gly Gly Gly Gly Ser'"

<400> 126

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

1 5 10 15

Gly Gly Gly Ser Gly Gly Gly Gly Ser

20 25

<210> 127

<211> 20

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

peptide"

<220>

<221> SITE

<222> (1)..(20)

<223> /note=" This sequence may contain 1-5 repeats

links 'Gly Gly Gly Ser'"

<400> 127

Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser

1 5 10 15

Gly Gly Gly Ser

20

<210> 128

<211> 7

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

protease cleavable sequence"

<400> 128

Gly Pro Leu Gly Val Arg Gly

1 5

<210> 129

<211> 8

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

protease cleavable sequence"

<400> 129

Ile Pro Val Ser Leu Arg Ser Gly

1 5

<210> 130

<211> 8

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

protease cleavable sequence"

<400> 130

Val Pro Leu Ser Leu Tyr Ser Gly

1 5

<210> 131

<211> 10

<212> Protein

<213> Unknown sequence

<220>

<221> source

<223> /note="Description of unknown sequence:

protease cleavable sequence"

<400> 131

Ser Gly Glu Ser Pro Ala Tyr Tyr Thr Ala

1 5 10

<210> 132

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

peptide"

<400> 132

Gly Gly Gly Gly Ser

1 5

<210> 133

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

peptide"

<400> 133

Gly Ser Gly Ser Gly Ser

1 5

<210> 134

<211> 13

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

peptide"

<400> 134

Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Thr

1 5 10

<210> 135

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

peptide"

<400> 135

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10 15

<210> 136

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

label 6xHis"

<400> 136

His His His His His

1 5

<210> 137

<211> 124

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence: synthetic

polypeptide"

<400> 137

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Gly Gly Gly Gly Gly Leu Asp Gly Asn

35 40 45

Glu Glu Pro Gly Gly Leu Glu Trp Val Ser Ser Ile Ser Gly Ser Gly

50 55 60

Arg Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser

65 70 75 80

Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg

85 90 95

Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser Leu Ser

100 105 110

Val Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<---

Claims (19)

1. Conditionally active IL-2 containing a fusion polypeptide of the formula: [A]-[L1]-[B]-[L2]-[D] or [A]-[L1]-[D]-[L2]-[B] or [D]-[L2]-[B]-[L1]-[A] or [B]-[L2]-[D]-[L1]-[A] or [D]-[L1]-[B]-[L1]-[A] or [B]-[L1]-[D]-[L1]-[A] or [B]-[L1]-[A]-[L1]-[D] or [D]-[L1]-[A]-[L1]-[B] Where [A] is an interleukin 2 (IL-2) polypeptide; [B] is a half-life extending element, wherein the half-life extending element is human serum albumin or an antigen-binding polypeptide that binds to serum albumin, or an immunoglobulin Fc; each of [L1] and [L2] is independently a polypeptide linker, wherein [L1] is a protease-cleavable polypeptide linker that comprises at least one sequence that is a cleavable protease, and [L2] is optionally a protease-cleavable polypeptide linker, and wherein the protease-cleavable linker [L2] comprises at least one sequence that is a cleavable protease, wherein for each of [L1] and [L2], independently, the protease is selected from the group consisting of kallikrein, thrombin, chymase, carboxypeptidase A, elastase, PR-3, granzyme M, calpain, matrix metalloproteinase (MMP), fibroblast activation protein (FAP), ADAM metalloproteinase, plasminogen activator, cathepsin, tryptase, and surface tumor cell proteases; [D] is an IL-2 blocking fragment, wherein the IL-2 blocking fragment is an antibody or antigen-binding fragment thereof that binds to IL-2 or a ligand-binding domain or cognate receptor fragment for an IL-2 polypeptide; and wherein the conditionally active IL-2 has an attenuated IL-2 receptor activating activity, wherein the IL-2 receptor activating activity of the fusion polypeptide is at least about 10-fold less than the IL-2 receptor activating activity of a polypeptide that comprises an IL-2 polypeptide obtained by cleavage of a protease cleavable linker L1, or, when L2 is protease cleavable, by cleavage of both L1 and L2, and wherein the IL-2 receptor activating activity is assessed using a CTLL-2 proliferation assay, a phospho-STAT ELISA, a HEK Blue reporter cell assay, with equivalent molar amounts of the IL-2 polypeptide and the fusion polypeptide. 2. Conditionally active IL-2 according to claim 1, characterized in that the blocking fragment of IL-2 inhibits the activation of the IL-2 receptor alpha/beta/gamma (IL-2Rαβγ) and the IL-2 receptor beta/gamma (IL-2Rβγ) by the IL-2 polypeptide in the uncleaved fusion polypeptide. 3. Conditionally active IL-2 according to claim 1, characterized in that [L2] is a protease-cleavable polypeptide linker. 4. Conditionally active IL-2 according to claim 1, characterized in that the IL-2 blocking fragment non-covalently binds to the IL-2 polypeptide. 5. The conditionally active IL-2 of claim 1, wherein the IL-2 blocking fragment is an antibody fragment that binds to the IL-2 polypeptide, and the antibody fragment is a single-domain antibody, Fab, or scFv. 6. Conditionally active IL-2 according to claim 1, characterized in that the cathepsin is cathepsin B, cathepsin C, cathepsin D, cathepsin E, cathepsin K, cathepsin L or cathepsin G. 7. The conditionally active IL-2 of claim 1, wherein the matrix metalloproteinase (MMP) is MMP1, MMP2, MMP3, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13 or MMP14. 8. A method for treating a tumor, comprising administering to a subject in need thereof an effective amount of conditionally active IL-2 according to any one of claims 1-7. 9. Use of conditionally active IL-2 according to any one of claims 1-7 for treating a tumor in a subject in need thereof. 10. A pharmaceutical composition for treating a tumor in a subject in need thereof, comprising as an active ingredient a conditionally active IL-2 according to any one of claims 1-7.