Epitopes of regulatory T cell surface antigens and antibodies specifically binding thereto

Abstract

The present invention relates to epitopes of leucine-rich immunoglobulin-like domain 1 (Lrig-1), an antigen present on the surface of regulatory T cells, and antibodies or antigen-binding fragments that specifically bind thereto.

Description

Technical Field

[0001] The present invention relates to an epitope of the leucine-rich and immunoglobulin-like domain 1 (Lrig-1) protein, which is an antigen present on the surface of regulatory T cells, and an antibody or antigen-binding fragment that specifically binds thereto. Background Art

[0002] One of the most important characteristics of all normal individuals is the ability to recognize and eliminate non-self antigens while simultaneously not generating a harmful reaction to antigenic substances that constitute the self. This inability of a living organism to respond to self-antigens is called immunological anergy or tolerance. Self-tolerance occurs through the elimination of lymphocytes that may possess specific receptors for self-antigens, or through the inactivation of the body's ability to respond after exposure to self-antigens. When there are problems in inducing or maintaining self-tolerance, an immune response to self-antigens occurs, and the resulting diseases are called autoimmune diseases.

[0003] In order to treat autoimmune diseases, Gershon first introduced and proposed the concept of suppressor T cells in the early 1970s, which suggested that there might be T cells that can control and inhibit the effector functions of conventional T cells (RK Gershon and K. Kondo, Immunology, 1970, 18:723-37). Since then, research has been conducted to clarify the biological characteristics and functions of regulatory T cells in many fields of immunology.

[0004] In this regard, regulatory T cells (Treg cells) have been reported to play an important role in naturally preventing excessive inflammation and immune responses; however, in the case of autoimmune diseases and chronic inflammatory diseases, the function and number of regulatory T cells are significantly reduced. Therefore, for patients with immune and inflammatory diseases, it is important to produce regulatory T cells at normal levels, which can be used as one of the treatment methods for these diseases.

[0005] To date, studies have been conducted on genes and proteins specifically present in regulatory T cells, and substances such as CD25, CTLA4, CD62L, CD38, CD103, GITR, and CD45RB have been proposed as possible markers. However, no gene or protein can target regulatory T cells alone.

[0006] On the other hand, there are three hypervariable regions called complementarity determining regions (hereinafter referred to as "CDRs") and four framework regions. CDRs are mainly used to bind to epitopes on antigens. The CDRs of each chain are usually referred to as CDR1, CDR2 and CDR3 starting from the N-terminus, and are also distinguished by the chain on which the specific CDR is located. Summary of the Invention

[0007] Technical issues

[0008] The present invention aims to provide an epitope of a leucine-rich and immunoglobulin-like domain 1 (Lrig-1) protein, which exists on the surface of regulatory T cells (Treg cells).

[0009] Another object of the present invention is to provide an antibody or antigen-binding fragment that can specifically bind to the epitope.

[0010] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, which comprises an antibody or antigen-binding fragment that can specifically bind to the epitope.

[0011] Another object of the present invention is to provide a method for preventing or treating cancer using an antibody or antigen-binding fragment that can specifically bind to the epitope.

[0012] However, the technical problems to be achieved by the present invention are not limited to the above problems, and those skilled in the art will clearly understand other problems not mentioned based on the following description.

[0013] Solution to the problem

[0014] The present invention relates to an epitope of a leucine-rich and immunoglobulin-like domain 1 (Lrig-1) protein, or an antibody or antigen-binding fragment that specifically binds to the epitope.

[0015] For the purposes of the present invention, "Lrig-1 protein" refers to a transmembrane protein present on the surface of regulatory T cells. It consists of a leucine-rich repeat (LRR) sequence, three immunoglobulin-like domains on the extracellular or luminal side, a transmembrane sequence, and a cytoplasmic tail. The LRIG gene family includes LRIG1, LRIG2, and LRIG3, and their amino acid sequences are highly conserved. The LRIG1 gene is highly expressed in normal skin and can also be expressed in basal and hair follicle cells to regulate the proliferation of epithelial stem cells. Therefore, the LRIG1 gene plays an important role in maintaining epidermal homeostasis, and loss of the LRIG1 gene may lead to the development of psoriasis or skin cancer. It has been reported that partial ablation of chromosome 3p14.3, where LRIG1 is located, may lead to the development of cancer cells. Indeed, LRIG1 expression has been shown to be significantly reduced in renal cell carcinoma and cutaneous squamous cell carcinoma. Recently, it has also been found that Lrig-1 is expressed in only approximately 20% to 30% of cancers. On the other hand, for the purposes of the present invention, the Lrig-1 protein may be, but is not limited to, a protein present in humans or mice.

[0016] In one embodiment of the present invention, the Lrig-1 protein may be an Lrig-1 protein derived from mammals, including primates such as humans and monkeys, and rodents such as mice and rats.

[0017] In one embodiment of the present invention, the Lrig-1 protein may be the human Lrig-1 protein represented by SEQ ID NO: 1, which may be, but is not limited to, encoded by the nucleic acid sequence represented by SEQ ID NO: 2 (see Table 1).

[0018] Table 1

[0019]

[0020]

[0021] In another embodiment of the present invention, the Lrig-1 protein may be, but is not limited to, the mouse-derived Lrig-1 protein represented by SEQ ID NO: 3 (see Table 2).

[0022] Table 2

[0023]

[0024] According to one embodiment of the present invention, there is provided an epitope as an epitope of Lrig-1 protein, which comprises a polypeptide consisting of an amino acid sequence represented by Formula 1,

[0025] [Formula 1]

[0026] Lx 1 Lx 2 x 3 N

[0027] In formula 1, x 1 to x 3 Each of the amino acids may be independently a neutral amino acid, an acidic amino acid, a basic amino acid, or an aromatic amino acid. Here, the neutral amino acid may be glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), serine (S), or threonine (T); the acidic amino acid may be aspartic acid (D), glutamic acid (E), asparagine (N), or glutamine (Q); the basic amino acid may be lysine (K), arginine (R), or histidine (H); and the aromatic amino acid may be phenylalanine (F) or tyrosine (Y).

[0028] In one embodiment of the present invention, x 1 to x 3Each may be independently an amino acid selected from the group consisting of asparagine (N), aspartic acid (D), serine (S), tyrosine (Y), arginine (R), phenylalanine (F), lysine (K), histidine (H), leucine (L), valine (V), threonine (T), alanine (A), glutamine (Q), glutamic acid (E) and glycine (G).

[0029] In another embodiment of the present invention, x 1 may be an amino acid selected from the group consisting of asparagine (N), phenylalanine (F), aspartic acid (D), lysine (K), histidine (H), valine (V), arginine (R) and threonine (T); x 2 may be an amino acid selected from the group consisting of serine (S), glutamine (Q), alanine (A), asparagine (N), glutamic acid (E), aspartic acid (D), phenylalanine (F) and glycine (G); x 3 It may be an amino acid selected from the group consisting of tyrosine (Y), histidine (H), glycine (G), arginine (R), asparagine (N), leucine (L), lysine (K) and phenylalanine (F).

[0030] In another embodiment of the present invention, x 1 to x 3 Each may be independently an amino acid selected from the group consisting of asparagine (N), aspartic acid (D), serine (S), tyrosine (Y) and arginine (R).

[0031] In yet another embodiment of the present invention, x 1 Can be asparagine (N) or aspartic acid (D); x 2 can be serine (S) or asparagine (N); and x 3 It can be tyrosine (Y) or arginine (R).

[0032] In the present invention, the epitope includes a polypeptide consisting of the amino acid sequence represented by Formula 1, and may consist of 10 to 20-mers, preferably 10 to 15-mers, and more preferably 11 to 14-mers.

[0033] In the present invention, the polypeptide consisting of the amino acid sequence represented by Formula 1 in the epitope may be located at positions 3 to 6, preferably positions 4 to 6, and more preferably position 5, from the N-terminus of the epitope.

[0034] As an embodiment of the present invention, the polypeptide composed of the amino acid sequence represented by Formula 1 can be represented by, but not limited to, any one of the amino acid sequences of SEQ ID NOs: 4 to 17 in Table 3 below:

[0035] [Table 3]

[0036]

[0037]

[0038] As another embodiment of the present invention, the epitope can be represented by, but is not limited to, the amino acid sequence of SEQ ID NO: 18 or 20.

[0039] According to another embodiment of the present invention, an epitope as an epitope of Lrig-1 protein is provided, which comprises a polypeptide represented by any one of the amino acid sequences of SEQ ID NOs: 18 to 29 in Table 4 below:

[0040] [Table 4]

[0041] SEQ ID NO Sequence information SEQ ID NO: 18 WTRSLNLSYNKL SEQ ID NO: 19 TEVRNTCFPHGPPI SEQ ID NO:20 RLTQLDLNRNRIR SEQ ID NO:21 DLNRNRIRLIEGLTF SEQ ID NO:22 NSIARIHRKGW SEQ ID NO:23 WLPPWLIGRMLQAF SEQ ID NO:24 RQVTFGHEGRY SEQ ID NO:25 FGHEGRYQCVITNHF SEQ ID NO:26 RLTVNVLPSFTKTPH SEQ ID NO:27 RRMHVMPDDDVFF SEQ ID NO:28 FFITDVKIDDAGVYS SEQ ID NO:29 KGDRPLSLTERHH

[0042] According to another embodiment of the present invention, a nucleic acid molecule encoding the epitope provided by the present invention is provided.

[0043] As known to those skilled in the art, nucleic acid molecules of the present invention comprise all nucleic acid molecules that obtain by translating the amino acid sequence of polypeptide provided by the invention into polynucleotide sequence.Therefore, can prepare various polynucleotide sequences by open reading frame (ORF), and all these polynucleotide sequences are also included in nucleic acid molecules of the present invention.

[0044] According to another embodiment of the present invention, an expression vector is provided, into which the isolated nucleic acid molecule provided by the present invention is inserted.

[0045] In the present invention, "vector" is a nucleic acid molecule capable of transporting another nucleic acid connected thereto. One type of vector is a "plasmid", which refers to a circular double-stranded DNA to which other DNA segments can be connected. Another type of vector is a phage vector. Another type of vector is a viral vector, in which other DNA segments can be connected to the viral genome. Certain vectors are capable of autonomous replication in the host cell into which they are introduced (for example, bacterial vectors with bacterial replication origins are episomal mammalian vectors). After being introduced into the host cell, other vectors (for example, non-episomal mammalian vectors) can be integrated into the genome of the host cell and therefore replicated together with the host genome. In addition, certain vectors can guide the expression of genes operably connected thereto. Such vectors are referred to herein as "recombinant expression vectors" or simply "expression vectors". Generally, expression vectors that can be used for recombinant DNA technology are generally in the form of plasmids. In this manual, "plasmid" and "vector" can be used interchangeably because plasmids are the most commonly used vector forms.

[0046] Specific examples of expression vectors in the present invention can be selected from, but are not limited to, the following groups: commercially widely used pCDNA vectors, F, R1, RP1, Col, pBR322, ToL, and Ti vectors; cosmids; bacteriophages such as λ, λ-shaped, M13, Mu, Pl, P22, Qμ, T-even, T2, T3, and T7; and plant viruses. Any known expression vector can be used as an expression vector in the present invention, and the expression vector is selected based on the properties of the target host cell. The vector can be introduced into the host cell via calcium phosphate transfection, viral infection, DEAE-dextran-mediated transfection, lipofection, or electroporation. However, the present invention is not limited thereto, and those skilled in the art can adopt and use an introduction method appropriate for the expression vector and host cell being used. The vector can be introduced into the host cell via calcium phosphate transfection, viral infection, DEAE-dextran-mediated transfection, lipofection, or electroporation. However, the present invention is not limited thereto, and those skilled in the art can adopt and use an introduction method appropriate for the expression vector and host cell being used. The vector preferably contains at least one selectable marker. Yet, the present invention is not limited thereto, and depending on whether product is produced, a vector that does not comprise a selection marker can be used for selection. Selectable markers are selected according to target host cells, which are accomplished using methods known to those skilled in the art, and therefore the present invention is not limited thereto.

[0047] To facilitate purification of the nucleic acid molecules of the present invention, a tag sequence can be inserted into and fused to an expression vector. Tags include, but are not limited to, hexahistidine tags, hemagglutinin tags, myc tags, or marker tags, and any tag known to those skilled in the art that facilitates purification can be used in the present invention.

[0048] According to another embodiment of the present invention, a host cell line transfected with the expression vector provided by the present invention is provided.

[0049] In the present invention, "host cell" includes a single cell or cell culture that can be or has been a recipient of a vector for incorporating a polypeptide insert. Host cells include the progeny of a single host cell, and the progeny are not necessarily completely identical (in morphology or genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation. Host cells include cells transfected in vivo with a polynucleotide herein.

[0050] In the present invention, the host cell may include a cell of mammalian, plant, insect, fungal or cell origin, and may be, for example, a bacterial cell such as Escherichia coli, Streptomyces, Salmonella typhimurium; a fungal cell such as a yeast cell and Pichia pastoris; an insect cell such as a fruit fly and Spodoptera frugiperda Sf9 cell; an animal cell such as a CHO (Chinese hamster ovary) cell, SP2 / 0 (mouse myeloma), a human lymphoblastoid cell, COS, NSO (mouse myeloma), 293T, Bowes melanoma cell, HT-1080, baby hamster kidney (BHK) cell, human embryonic kidney (HEK) cell or PERC.6 (human retinal cell); or a plant cell. However, the host cell is not limited thereto, and any cell known to those skilled in the art that can be used as a host cell line is available.

[0051] According to yet another embodiment of the present invention, an antibody or antigen-binding fragment that specifically binds to the epitope of the present invention is provided.

[0052] In addition, the antibody according to the present invention specifically binds to an epitope, which includes a polypeptide consisting of an amino acid sequence represented by Formula 1, or an epitope of a polypeptide represented by any one of the amino acid sequences of SEQ ID NOs: 18 to 29, in the Lrig-1 protein present on regulatory T cells, thereby inhibiting the function of regulatory T cells and maintaining or increasing the activity of effector T cells, thereby effectively inhibiting the growth of cancer cells, especially solid cancer cells.

[0053] As used herein, the term "cancer" refers to or indicates a physiological condition characterized by the growth of mammalian cells that is not regulated in a typical manner. The cancer to be prevented, improved, or treated in the present invention may be a solid tumor formed by aggregates caused by abnormal cell growth in a solid organ, and may be, but is not limited to, gastric cancer, liver cancer, glioma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, lung cancer, etc., depending on the location of the solid organ.

[0054] In the present invention, an antibody as a full-length antibody or as a portion of an antibody has the ability to bind to the Lrig-1 protein, and includes any antibody fragment that binds to the Lrig-1 antigenic determinant site in a competitive manner with the binding molecule of the present invention.

[0055] As used herein, "antibody" refers to a protein molecule that acts as a receptor for specific antigen recognition, including immunoglobulin molecules that immunoreact with a specific antigen. For the purposes of the present invention, the antigen may be the Lrig-1 protein present on the surface of regulatory T cells. Preferably, the antibody specifically recognizes the leucine-rich region or immunoglobulin-like domain of the Lrig-1 protein, but is not limited thereto.

[0056] In the present invention, "immunoglobulin" has a heavy chain and a light chain, and each of the heavy chain and the light chain comprises a constant region and a variable region. The variable region of each of the light chain and the heavy chain comprises three hypervariable regions (hereinafter referred to as "CDRs") called complementarity determining regions and four framework regions. CDRs are mainly used to bind to epitopes on antigens. The CDRs of each chain are usually referred to as CDR1, CDR2 and CDR3 starting from the N-terminus, and are also distinguished by the chain in which the specific CDRs are located.

[0057] In the present invention, a "full-length antibody" has a structure consisting of two full-length light chains and two full-length heavy chains, wherein each light chain is linked to a heavy chain by a disulfide bond, and includes IgA, IgD, IgE, IgM, and IgG. IgG includes IgG1, IgG2, IgG3, and IgG4 as its subtypes.

[0058] In addition, as used herein, "antigen-binding fragment" refers to a fragment having an antigen-binding function, and examples of antigen-binding fragments include (i) a Fab fragment consisting of a light chain variable region (VL), a heavy chain variable region (VH), a light chain constant region (CL) and a heavy chain constant region 1 (CH1); (ii) an Fd fragment consisting of the VH and CH1 domains; (iii) an Fv fragment consisting of the VL and VH domains of a single antibody arm; (iv) a dAb fragment consisting of a VH domain (Ward, ES et al., Nature 2012). 341: 544-546 (1989)); (v) isolated CDR regions; (vi) F(ab')2 fragments, which are bivalent fragments comprising two linked Fab fragments; (vii) single-chain Fv molecules (scFv) in which the VH domain and the VL domain are linked by a peptide linker that allows the two domains to associate to form an antigen-binding site; (viii) bispecific single-chain Fv dimers; and (ix) diabodies, which are multivalent or multispecific fragments constructed by gene fusion. In the case of using proteolytic enzymes such as papain or pepsin, antigen-binding fragments can be obtained as Fab or F(ab')2 fragments and can be produced by genetic recombination techniques.

[0059] In addition, in the present invention, the antibody can be, but is not limited to, a monoclonal antibody, a polyclonal antibody, a chimeric antibody, a humanized antibody, a bivalent bispecific molecule, a minibody, a domain antibody, a bispecific antibody, a mimetic antibody, a diabody, a triabody or a tetrabody or a fragment thereof.

[0060] Additionally, as used herein, "monoclonal antibody" refers to an antibody molecule of single molecular composition obtained from a population of substantially identical antibodies and that displays a single binding specificity and affinity for a particular epitope.

[0061] In the present invention, a "chimeric antibody" is an antibody obtained by recombining the variable region of a mouse antibody with the constant region of a human antibody, and has a greatly improved immune response compared to a mouse antibody.

[0062] In addition, as used herein, "humanized antibody" refers to an antibody obtained by modifying the protein sequence of an antibody derived from a non-human species so that the protein sequence is similar to a naturally occurring antibody variant in humans. For example, humanized antibodies can be prepared as follows. Mouse-derived CDRs can be recombined with human antibody-derived FRs to prepare humanized variable regions, and humanized variable regions can be recombined with preferably human antibody constant regions to prepare humanized antibodies.

[0063] As used herein, "binding" or "specific binding" refers to the affinity of an antibody or antibody composition herein for an antigen. In antigen-antibody binding, "specific binding" is distinguished from nonspecific background binding, typically with a dissociation constant (Kd) of less than 1×10 -5 M, less than 1×10 -6 M, or less than 1×10 -7 In the case of M. Specific binding can be detected by methods known in the art, such as ELISA, surface plasmon resonance (SPR), immunoprecipitation, and coprecipitation, including appropriate controls that can distinguish between nonspecific binding and specific binding.

[0064] The antibody or antigen-binding fragment of the present invention can exist in a multimeric form, such as a dimer, trimer, tetramer or pentamer, which comprises the antigen-binding ability of at least a portion of a monomer. Such multimers also include homopolymers or heteropolymers. Antibody multimers contain a large number of antigen-binding sites and therefore have excellent antigen-binding ability compared to monomers. Antibody multimers are also easily used to produce multifunctional (i.e., bifunctional, trifunctional, tetrafunctional, etc.) antibodies.

[0065] As used herein, "multifunctional" refers to an antibody or antigen-binding fragment having two or more activities or functions (e.g., antigen binding ability, enzyme activity, and ligand or receptor binding ability). For example, the antibodies of the present invention can be bound to polypeptides having enzyme activity, such as luciferase, acetyltransferase, and β-galactosidase. Multifunctional antibodies also include antibodies in the form of multivalent or multispecific (i.e., bispecific, trispecific, etc.).

[0066] According to another embodiment of the present invention, an antibody-drug conjugate (ADC) is provided, which comprises the antibody or antigen-binding fragment provided by the present invention and a drug.

[0067] As used herein, "antibody-drug conjugate (ADC)" refers to a form in which a drug and an antibody are chemically linked to each other without reducing the biological activities of the antibody and the drug. In the present invention, an antibody-drug conjugate refers to a form in which the drug is bound to the amino acid residue at the N-terminus of the heavy chain and / or light chain of the antibody, particularly a form in which the drug is bound to the N-terminus of the heavy chain and / or light chain of the antibody and an α-amino group.

[0068] As used herein, "drug" refers to any substance that has a certain biological activity on cells, including DNA, RNA, or peptides. The drug may be in a form containing a reactive group capable of reacting with and cross-linking an α-amino group, or may include a form containing a reactive group capable of reacting with and cross-linking an α-amino group and a linker attached thereto.

[0069] In the present invention, examples of reactive groups capable of reacting and cross-linking with α-amino groups are not particularly limited in type, as long as the reactive group can react and cross-link with the α-amino group at the N-terminus of the heavy chain or light chain of the antibody. Reactive groups include all types of groups known in the art that react with amino groups. Reactive groups can be, for example, any of isothiocyanates, isocyanates, acyl azides, NHS esters, sulfonyl chlorides, aldehydes, glyoxal, epoxides, ethylene oxides, carbonates, aryl halides, imido esters, carbodiimides, acid anhydrides, and fluorophenyl esters, but are not limited thereto.

[0070] In the present invention, the antibody-drug conjugate comprises an antibody or antigen-binding fragment that specifically binds to the epitope of the present invention in the Lrig-1 protein, i.e., an epitope of a polypeptide consisting of the amino acid sequence represented by Formula 1 or an epitope of a polypeptide represented by any one of the amino acid sequences of SEQ ID NOs: 18 to 29, wherein the drug can be a drug capable of treating cancer, a disease targeted by the Lrig-1 antibody, i.e., an anticancer drug.

[0071] In the present invention, the anticancer agent may include any drug without limitation, as long as the drug is used to prevent, improve or treat cancer. The anticancer agent may be, for example, selected from the group consisting of nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semaxanib, bosutinib, axitinib, cediranib, retautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, white mistletoe, Asparaginase, retinoic acid, hydroxyurea, dasatinib, estramustine, gemtuzumab, octopirox, ibritumomab tiuxetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine, docetaxel, pemetrexed, tegafur, capecitabine, gimeracil, oteracil, azacitidine, methotrexate, uracil, cytarabine, fluorouracil doxycycline, fludarabine, enocitabine, flutamide, capecitabine, decitabine, mercaptopurine, thioguanine, cladribine, carmofur, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vinblastine, idarubicin, mitomycin, bleomycin, dactinomycin, pirarubicin, aclarubicin, peplomycin, temsirolimus, temozolomide, busulfan The anticancer agent includes, but is not limited to, cyclophosphamide, melphalan, hexamethylmelamine, dacarbazine, thiotepa, nimustine, chlorambucil, dibromodimethoate, folinic acid, tretinoin, exemestane, aminoglutethimide, anagrelide, olaparib, vinorelbine, fadrozole, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine, 5FU, vorinostat, entinostat, and carmustine. However, the anticancer agent is not limited thereto.

[0072] In the present invention, cancer may be a solid tumor formed by an aggregate caused by abnormal growth of cells in a solid organ, and specific examples thereof may include, but are not limited to, gastric cancer, liver cancer, glioma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, lung cancer, etc., depending on the location of the solid organ.

[0073] According to another embodiment of the present invention, a pharmaceutical composition for preventing or treating cancer is provided, which comprises as an active ingredient the antibody or antigen-binding fragment provided by the present invention or the antibody-drug conjugate (ADC) provided by the present invention.

[0074] In the present invention, the antibody or antigen-binding fragment contained as an active ingredient in the pharmaceutical composition, or the antibody-drug conjugate obtained by binding to a drug, specifically binds to an epitope in the Lrig-1 protein present on regulatory T cells, including an epitope of a polypeptide consisting of the amino acid sequence represented by Formula 1, or an epitope of a polypeptide represented by any one of the amino acid sequences of SEQ ID NOs: 18 to 29, thereby inhibiting the function of regulatory T cells and maintaining or increasing the activity of effector T cells, thereby effectively inhibiting the growth of cancer cells, particularly solid cancer cells.

[0075] In the present invention, cancer may be a solid tumor formed by an aggregate caused by abnormal growth of cells in a solid organ, and specific examples thereof may include, but are not limited to, gastric cancer, liver cancer, glioma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, lung cancer, etc., depending on the location of the solid organ.

[0076] Meanwhile, in the present invention, "prevention" may include but is not limited to any action of blocking disease symptoms or inhibiting or delaying symptoms using the pharmaceutical composition of the present invention.

[0077] Furthermore, in the present invention, "treatment" may include, but is not limited to, any action of using the pharmaceutical composition of the present invention to improve or beneficially change the symptoms of a disease.

[0078] In the present invention, the pharmaceutical composition may be characterized as being in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be characterized as being targeted to humans.

[0079] In the present invention, the pharmaceutical composition can be prepared and used in the form of oral preparations, such as powders, granules, capsules, tablets and aqueous suspensions, external preparations, suppositories and sterile injections according to conventional methods. However, the pharmaceutical composition is not limited thereto. The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. As pharmaceutically acceptable carriers, binders, glidants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavorings, etc. can be used for oral administration; buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc. can be mixed for injection; alkalis, excipients, lubricants, preservatives, etc. can be used for topical administration. The preparation of the pharmaceutical composition of the present invention can be prepared by mixing with the pharmaceutically acceptable carriers described above in various ways. For example, for oral administration, the pharmaceutical composition can be prepared in the form of tablets, lozenges, capsules, elixirs, suspensions, syrups, implants (wafers), etc. For injection, the pharmaceutical composition can be formulated into unit dose ampoules or multiple doses. Alternatively, the pharmaceutical composition can be formulated into a solution, suspension, tablet, capsule, sustained-release formulation, or the like.

[0080] Meanwhile, as examples of carriers, diluents or excipients suitable for preparing the preparation, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, etc. can be used. In addition, fillers, anticoagulants, lubricants, wetting agents, flavors, emulsifiers, preservatives, etc. can be further included.

[0081] In the present invention, the administration route of the pharmaceutical composition includes but is not limited to oral, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal routes. Oral or parenteral administration is preferred.

[0082] In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intracapsular, intrasternal, intradural, intralesional and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention can also be used in the form of suppositories for rectal administration.

[0083] The pharmaceutical composition of the present invention can be widely changed according to many factors, including the activity of the certain compound used, the patient's age, body weight, overall health, sex, diet, administration time, route of administration, excretion rate, drug combination and the severity of certain diseases to be prevented or treated. The dosage of the pharmaceutical composition can change according to the patient's condition, body weight, the severity of the disease, drug form, route of administration and duration, and can be suitably selected by those skilled in the art. The pharmaceutical composition can be used in an amount of 0.0001 to 50mg / kg or 0.001 to 50mg / kg every day. Administration can be once a day or several times a day. This dosage is not intended to limit the scope of the present invention in any way. The pharmaceutical composition of the present invention can be formulated into the form of pills, sugar-coated tablets, capsules, liquids, gels, syrups, slurries or suspensions.

[0084] According to another embodiment of the present invention, a method for preventing or treating cancer is provided, comprising the step of administering to an individual an antibody or antigen-binding fragment according to the present invention, or an antibody-drug conjugate (ADC) according to the present invention.

[0085] The antibodies or antigen-binding fragments of the present invention and the antibody-drug conjugates of the present invention specifically bind to an epitope in the Lrig-1 protein present on regulatory T cells, wherein the epitope includes a polypeptide consisting of an amino acid sequence represented by Formula 1, or the epitope includes a polypeptide represented by any one of the amino acid sequences of SEQ ID NOs: 18 to 29, thereby inhibiting the function of regulatory T cells and maintaining or increasing the activity of effector T cells, thereby effectively inhibiting the growth of cancer cells, particularly solid cancer cells.

[0086] In the present invention, "subject" refers to an individual suspected of having cancer, and an individual suspected of having cancer refers to a mammal that has developed or is likely to develop the disease, such as humans, mice, and livestock. However, any individual treated with the antibody or antibody-drug conjugate of the present invention is included herein without limitation.

[0087] The methods of the present invention may include administering an antibody or antibody-drug conjugate in a pharmaceutically effective amount. The attending physician or veterinarian may determine an appropriate total daily dose within the scope of sound medical judgment, and the dose may be administered in one or more doses. However, for the purposes of the present invention, a specific therapeutically effective amount for a particular patient is preferably applied differently depending on various factors, including the type and extent of the response to be achieved, whether the specific composition is used with other agents, and optionally the patient's age, weight, general health, sex and diet, time of administration, route of administration, rate of secretion of the composition, duration of treatment, and drugs used concurrently or in combination with the specific composition, as well as similar factors well known in the medical field.

[0088] Meanwhile, the method for preventing or treating cancer may be, but is not limited to, combination therapy, which further includes administering a compound or substance having therapeutic activity against one or more cancer diseases.

[0089] In the present invention, "combination" is understood to mean simultaneous, separate or sequential administration. In the case of sequential or separate administration, the second component should be administered at a certain interval so that the beneficial effects of the combination are not lost.

[0090] In the present invention, the dosage of the antibody or antibody-drug conjugate can be, but is not limited to, about 0.0001 μg to 500 mg per kilogram of patient body weight.

[0091] In the present invention, cancer may be a solid tumor formed by an aggregate caused by abnormal growth of cells in a solid organ, and specific examples thereof may include, but are not limited to, gastric cancer, liver cancer, glioma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, lung cancer, etc., depending on the location of the solid organ.

[0092] Advantageous Effects of the Invention

[0093] According to the present invention, antibodies or antigen-binding fragments that specifically bind to the epitope of the Lrig-1 protein specifically bind to the epitope of the present invention present on regulatory T cells, thereby inhibiting the function of regulatory T cells and maintaining or increasing the activity of effector T cells, thereby effectively inhibiting the growth of cancer cells, especially solid cancer cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0095] Figure 1 The structure of the Lrig-1 protein according to one embodiment of the present invention is shown.

[0096] Figure 2 The structure of the Lrig-1 protein according to one embodiment of the present invention is shown.

[0097] Figure 3 The expression level of Lrig-1 mRNA according to one embodiment of the present invention is shown.

[0098] Figure 4 The expression level of Lrig-1 mRNA according to one embodiment of the present invention is shown.

[0099] Figure 5 The expression level of Lrig-1 mRNA according to one embodiment of the present invention is shown.

[0100] Figure 6 The expression levels of Lrig-1, Lrig-2, and Lrig-3 mRNA according to one embodiment of the present invention are shown.

[0101] Figure 7 The figures show the results obtained by comparing the expression levels of Lrig-1 protein in regulatory T cells and non-regulatory T cells according to one embodiment of the present invention.

[0102] Figure 8 FIG. 1 shows the expression of Lrig-1 protein on the surface of regulatory T cells according to one embodiment of the present invention.

[0103] Figure 9 Shown are the results obtained by analyzing the ability of antibodies (A7, C8, E7, G3, A8, B8, D9, and H6) to bind to the Lrig-1 protein in one embodiment of the present invention.

[0104] Figure 10 The present invention shows the results obtained by analyzing the mechanism of Lrig-1 protein-induced Stat3 phosphorylation in regulatory T cells regulated by Lrig-1 protein-specific monoclonal antibodies (A7, C8, E7, G3, A8, B8, D9, and H6) in one embodiment of the present invention.

[0105] Figure 11 The figure shows the experimental design for cancer treatment using Lrig-1 protein-specific monoclonal antibodies (A8, B8, D9, and H6) in one embodiment of the present invention.

[0106] Figure 12 The cancer therapeutic effects obtained by using Lrig-1 protein-specific monoclonal antibodies (A8, B8, D9, and H6) in one embodiment of the present invention are shown.

[0107] Figure 13 The graph shows the results of epitope mapping of the Lrig-1 protein using a 10 μg / ml monoclonal antibody (H6) on a microarray in one embodiment of the present invention.

[0108] Figure 14 The figures show the results of epitope mapping of the Lrig-1 protein using a 100 μg / ml monoclonal antibody (H6) on a microarray in one embodiment of the present invention.

[0109] Figure 15 The figures show the results of epitope mapping of the Lrig-1 protein using a monoclonal antibody (H6) on a microarray in one embodiment of the present invention. DETAILED DESCRIPTION

[0110] The present invention relates to an epitope of a leucine-rich and immunoglobulin-like domain 1 (Lrig-1) protein, or an antibody or antigen-binding fragment that specifically binds to the epitope.

[0111] According to one embodiment of the present invention, an epitope as an epitope of Lrig-1 protein is provided, which includes a polypeptide consisting of an amino acid sequence represented by Formula 1.

[0112] [Formula 1]

[0113] Lx 1 Lx 2 x 3 N

[0114] In formula 1, x 1 to x 3 Each of the amino acids may be independently a neutral amino acid, an acidic amino acid, a basic amino acid, or an aromatic amino acid. Herein, the neutral amino acid may be glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), serine (S), or threonine (T); the acidic amino acid may be aspartic acid (D), glutamic acid (E), asparagine (N), or glutamine (Q); the basic amino acid may be lysine (K), arginine (R), or histidine (H); and the aromatic amino acid may be phenylalanine (F) or tyrosine (Y).

[0115] In one embodiment of the present invention, x 1 to x 3 Each may be independently an amino acid selected from the group consisting of asparagine (N), aspartic acid (D), serine (S), tyrosine (Y), arginine (R), phenylalanine (F), lysine (K), histidine (H), leucine (L), valine (V), threonine (T), alanine (A), glutamine (Q), glutamic acid (E) and glycine (G).

[0116] In another embodiment of the present invention, x 1 may be an amino acid selected from the group consisting of asparagine (N), phenylalanine (F), aspartic acid (D), lysine (K), histidine (H), valine (V), arginine (R) and threonine (T); x 2 may be an amino acid selected from the group consisting of serine (S), glutamine (Q), alanine (A), asparagine (N), glutamic acid (E), aspartic acid (D), phenylalanine (F) and glycine (G); x 3 It may be an amino acid selected from the group consisting of tyrosine (Y), histidine (H), glycine (G), arginine (R), asparagine (N), leucine (L), lysine (K) and phenylalanine (F).

[0117] In another embodiment of the present invention, x 1 to x 3 Each may be independently an amino acid selected from the group consisting of asparagine (N), aspartic acid (D), serine (S), tyrosine (Y) and arginine (R).

[0118] In yet another embodiment of the present invention, x 1 Can be asparagine (N) or aspartic acid (D); x 2 Can be serine (S) or asparagine (N); x 3 It can be tyrosine (Y) or arginine (R).

[0119] In another embodiment of the present invention, the polypeptide consisting of the amino acid sequence represented by Formula 1 can be, but is not limited to, represented by any one of the amino acid sequences of SEQ ID NOs: 4 to 17.

[0120] According to another embodiment of the present invention, an epitope as an epitope of Lrig-1 protein is provided, which includes a polypeptide represented by any one of the amino acid sequences of SEQ ID NOs: 18 to 29 in Table 4.

[0121] According to another embodiment of the present invention, an antibody or antigen-binding fragment that specifically binds to an epitope of the present invention is provided.

[0122] According to another embodiment of the present invention, a pharmaceutical composition for preventing or treating cancer is provided, which comprises the antibody or antigen-binding fragment provided by the present invention as an active ingredient.

[0123] Hereinafter, the present invention will be described in more detail by way of examples. These examples are only used to describe the present invention in more detail, and it will be apparent to those skilled in the art that, according to the gist of the present invention, the scope of the present invention is not limited by these examples.

[0124] Example

[0125] [Preparation Example 1] T cell subtype cell culture

[0126] To determine whether Lrig-1 protein is expressed exclusively in regulatory T (Treg) cells, T cell subsets Th0, Th1, Th2, Th17, and iTreg were generated. Unlike naturally isolated nTreg, iTreg refers to cells artificially induced to differentiate in a culture medium with the following composition.

[0127] The primary T cells obtained from the mouse spleen were first isolated, and the RPMI1640 (Invitrogen Gibco, Grand Island, New York) nutrient medium containing 10% fetal bovine serum (FBS; HyClone, Logan, Utah) further contained the various components listed in Table 5 below, and cultured in a 37°C, 5% CO2 incubator for 72 hours to induce T cell subsets to differentiate into corresponding cells.

[0128] Table 5

[0129] Differentiated cells composition Th0 Anti-CD3, anti-CD28 Th1 IL-12, anti-IL-4 antibodies Th2 IL-4, anti-IFNβ Th17 IL-6, TGFβ, anti-IFNβ, anti-IL-4 iTreg IL-2, TGFβ

[0130] [Example 1] Structural Analysis of Lrig-1

[0131] The three-dimensional spatial structure of the extracellular domain of the Lrig-1 protein was predicted to generate antibodies specific to the Lrig-1 protein, which is a surface protein of regulatory T cells.

[0132] First, in order to predict the base sequence of the epitope, the tools of Uniprot (http: / / www.uniprot.org) and RCSB protein database (http: / / www.rcsb.org / pdb) were used to predict the three-dimensional structure of the extracellular domain (ECD) of Lrig-1 protein in order to identify the structure of ECD. Figure 1 and 2 Shown in.

[0133] like Figure 1As shown, there are a total of 15 leucine-rich regions, LRR1 to LRR15, in the Lrig-LRR domain of the extracellular domain of the Lrig-1 protein (amino acid sequence at positions 41-494). Each LRR domain consists of 23-27 amino acids, of which 3 to 5 leucines are present.

[0134] In addition, if Figure 2 As shown, there are three immunoglobulin-like domains in the amino acid sequence of positions 494 to 781 of the Lrig-1 protein in the extracellular domain of the Lrig-1 protein.

[0135] [Example 2] Identification of specific expression of Lrig-1 mRNA in regulatory T cells

[0136] The results verified whether Lrig-1 protein can be used as a specific biomarker for regulatory T cells.

[0137] For validation, CD4+ cells were isolated from mouse spleen using magnetic activated cell sorting (MACS). 4+ T cells. Subsequently, regulatory T (CD4 + CD 25+ T) cells and non-regulatory T (CD4 + CD 25- For each cell and the cells differentiated in Preparation Example 1, mRNA was extracted using Trizol, and then gDNA was removed from genomic RNA using a gDNA extraction kit (Qiagen) according to the manufacturer's protocol. The mRNA from which gDNA was removed was synthesized into cDNA using a BDsprint cDNA synthesis kit (Clontech).

[0138] Real-time polymerase chain reaction (RT PCR) was performed to quantitatively identify the expression level of Lrig-1 mRNA in cDNA.

[0139] Real-time polymerase chain reaction was performed using SYBR Green (Molecular Probes) according to the manufacturer's protocol using the primers shown in Table 6 below. The conditions were: 40 cycles of 95°C for 3 minutes, 61°C for 15 seconds, and 72°C for 30 seconds. Relative gene expression levels were calculated using the ΔCT method and normalized using HPRT. The results are shown in Figure 3-6 Shown in.

[0140] Table 6

[0141]

[0142] like Figure 3As shown, it can be seen that Lrig-1 is in the regulatory T (CD4 + CD25 + T) cells than non-regulatory T (CD4 + CD25 - The expression level of regulatory T cells was 18.1-fold higher than that of Lag3 and Ikzf4, markers of regulatory T cells.

[0143] In addition, if Figure 4 and 5 As shown, Lrig-1 mRNA is significantly more highly expressed in regulatory T cells compared with other types of immune cells, especially in naturally isolated regulatory T cells (nTreg) compared with inducible regulatory T (iTreg) cells.

[0144] In addition, if Figure 6 As shown, among Lrig-1, Lrig-2, and Lrig-3 corresponding to the Lrig family, Lrig-1 has the highest expression.

[0145] It can be seen from the above results that the Lrig-1 protein according to the present invention is specifically expressed in regulatory T cells, especially naturally occurring regulatory T cells.

[0146] [Example 3] Identification of the specific expression of Lrig-1 protein in regulatory T cells

[0147] We investigated whether Lrig-1 protein expressed from Lrig-1 mRNA is specifically expressed only in regulatory T cells.

[0148] FOXP3-RFP knock-in mice were used to generate FOXP3-RFP by coupling red fluorescent protein (RFP) to the FOXP3 promoter, a transcription factor specific for regulatory T cells. CD4 T cells were isolated from the spleen of mice using magnetic activated cell sorting (MACS) with CD4 beads. + T cells. Subsequently, RFP protein was used to separate them by fluorescence activated cell sorting (FACS) to obtain regulatory T (CD4 + RFP + T) cells and non-regulatory T (CD4 + RFP - T) cells. Each cell was stained with purchased Lrig-1 antibody, and the negative control was stained with an isotype-matched control antibody to detect the expression level of Lrig-1 using a fluorescence-activated cell sorter. Figure 7 Shown in.

[0149] like Figure 7As shown, non-regulatory T cells indicated by a dotted line showed almost the same Lrig-1 expression level as the negative control, whereas a large number of cells highly expressing Lrig-1 were present among the regulatory T cells.

[0150] From the above results, it can be seen that the Lrig-1 protein according to the present invention is specifically expressed in regulatory T cells.

[0151] [Example 4] Identification of the specific expression of Lrig-1 protein on the surface of regulatory T cells

[0152] From the perspective of becoming a target for cell therapy, Lrig-1 protein must be expressed on the surface of regulatory T cells, so that targeted therapy can be more effectively performed. We identified whether Lrig-1 protein is expressed on the surface of regulatory T cells.

[0153] Each differentiated T cell subset of Preparation Example 1 was stained using anti-CD4-APC and anti-Lrig-1-PE antibodies, and the expression level of Lrig-1 on the surface of each cell was measured using a fluorescence activated cell sorter (FACS). Figure 8 Shown in.

[0154] like Figure 8 As shown, the expression level of Lrig-1 in activated T cells, Th1 cells, Th2 cells, Th17 cells and naive T cells ranged from 0.77 to 15.3, while the expression level of Lrig-1 in differentiation-induced T cells (iTreg cells) was as high as 83.9.

[0155] From the above results, it can be seen that the Lrig-1 protein according to the present invention is not only specifically expressed in regulatory T (Treg) cells, but is also expressed at a higher level particularly on the surface of Treg cells.

[0156] [Production Examples 1 to 8] Production of Specific Monoclonal Antibodies to Lrig-1 Protein

[0157] Production of antibodies specific for the Lrig-1 protein according to the present invention The antibodies of the present invention are not generated by specifying a certain epitope, but are generated as antibodies capable of binding to any site on the Lrig-1 protein.

[0158] To produce antibodies, cells expressing the Lrig-1 protein were prepared. More specifically, a DNA fragment corresponding to SEQ ID NO: 2 and pcDNA (hygro) were cleaved with a cleavage enzyme, incubated at 37°C, and ligated to produce pcDNA into which the DNA sequence for the Lrig-1 protein was inserted. The resulting pcDNA, which contained the DNA sequence for the Lrig-1 protein, was then introduced into L cells via transfection, resulting in expression of the Lrig-1 protein on the cell surface.

[0159] Light and heavy chain amino acid sequences capable of binding to Lrig-1 expressed on the cell surface were selected from the human scFv library, resulting in a total of eight heavy and light chains.

[0160] The selected heavy and light chain amino acid sequences were fused to the mlgG2a Fc region to generate monoclonal antibodies. The sequences of the monoclonal antibodies are shown in Table 7 below.

[0161] [Table 7]

[0162]

[0163]

[0164]

[0165]

[0166] [Example 5] Evaluation of the binding ability of the antibodies of the present invention to Lrig-1 protein

[0167] In order to determine whether the monoclonal antibodies according to the present invention produced in Production Examples 1 to 8 recognize Lrig-1 well, each of the antibodies in Production Examples 1 to 8 was bound to L cells stably expressing Lrig-1. Then, a secondary antibody coupled to eFlour670 and capable of recognizing mouse antibodies was added thereto, and the binding ability of the monoclonal antibodies to the Lrig-1 protein was analyzed using FACS. The results were as follows: Figure 9 Shown in.

[0168] like Figure 9 As shown, all of the Lrig-1 protein-specific monoclonal antibodies according to the present invention (A7, A8, B8, C8, D9, E7, G3, and H6) were found to effectively recognize and bind to the Lrig-1 protein present on the surface of L cells.

[0169] [Example 6] Regulation of signal transduction pathways in regulatory T cells by the antibodies of the present invention

[0170] In order to analyze how the monoclonal antibodies according to the present invention produced in Production Examples 1 to 8 affect the signal transduction pathway in regulatory T cells through the Lrig-1 protein, Lrig-1 present on the surface of regulatory T cells was stimulated by treating the regulatory T cells with the antibodies of Production Examples 1 to 8, and then the tyrosine phosphorylation level of the Stat3 protein present in the stimulated regulatory T cells was analyzed by phosphotyrosine immunoblotting. Figure 10 Shown in.

[0171] like Figure 10As shown in the results, it was found that the Lrig-1 protein-specific monoclonal antibodies according to the present invention (A7, C8, E7, and G3) increased the phosphorylation of Stat3 to the same level as that of Th17 cells. On the other hand, it was found that the Lrig-1 protein-specific monoclonal antibodies according to the present invention (A8, B8, D9, and H6) continued to maintain and reduce the phosphorylation of Stat3 at the same level as that of iTreg cells.

[0172] [Example 7] Cancer therapeutic effects of A8, B8, D9 and H6 antibodies

[0173] In order to identify the therapeutic effects of the monoclonal antibodies (A8, B8, D9 and H6) according to the present invention produced in Production Examples 5 to 8 on solid cancers, Figure 11 As shown, B16F10 melanoma cells were cultured at 3×10 5 The amount of cells was subcutaneously injected into the back of the mice, and then the antibodies of Production Examples 5 to 8 were intraperitoneally injected into the mice at an amount of 200 μg on days 4, 8 and 12. After the melanoma cells were transplanted, the changes in tumor volume over time were measured, and the results are shown in Figure 12 middle.

[0174] like Figure 12 As shown, it was found that a significant reduction in melanoma tumor size was observed in the case of treatment with the Lrig-1 protein-specific monoclonal antibodies (A8, B8, D9 and H6) of the present invention, compared to the negative control that did not receive antibody treatment.

[0175] From these results, it can be seen that the specific monoclonal antibody against Lrig-1 protein according to the present invention inhibits the growth of various solid cancer cells, thereby effectively preventing, improving or treating such cancers.

[0176] [Example 8] Determination of epitopes in Lrig-1 protein according to the present invention

[0177] In order to find the epitope to which the H6 antibody of Production Example 8 binds, wherein the epitope exists in each LRR domain and Ig-like domain of the Lrig-1 protein present on the surface of regulatory T cells, the following experiment was performed.

[0178] 1. Experimental Preparation

[0179] (1) Microarray

[0180] The Lrig-1 protein was extended at its C- and N-termini with GSGSGSG linkers to prevent peptide truncation. The extended antigen sequence was translated into 15 amino acids with a peptide-peptide overlap of 14 amino acids. The resulting Lrig-1 peptide microarray contained 1,091 different peptides, printed in duplicate and framed with a control peptide of another HA (YPYDVPDYAG, 102 spots).

[0181] (2) Experimental materials

[0182] Sample: H6 monoclonal antibody of Production Example 8

[0183] Wash buffer: PBS, pH 7.4, containing 0.05% Tween 20 (3x, 10 seconds after each incubation)

[0184] Blocking buffer: Rockland Blocking Buffer MB-070 (30 minutes before the first measurement)

[0185] Incubation buffer: Wash buffer containing 10% blocking buffer

[0186] Assay conditions: Antibody concentrations in the incubation buffer were adjusted to 1 μg / ml, 10 μg / ml, and 100 μg / ml and incubated at 4 °C for 16 h with stirring at 140 rpm.

[0187] Secondary antibody: goat anti-human IgG (H+L) DyLight 680 (0.2 μg / ml); staining in incubation buffer at room temperature for 45 minutes

[0188] Control antibody: mouse monoclonal anti-HA (12CA5) DyLight800 (0.5 μg / ml); staining in incubation buffer at room temperature for 45 minutes

[0189] Scanner: LI-COR Odyssey Imaging System; scanning offset 0.65 mm, resolution 21 μm, scanning intensity 7 / 7 (red = 700 nm / green = 800 nm)

[0190] 2. Experimental Methods

[0191] Lrig-1 peptide microarray copies were prestained in incubation buffer with secondary and control antibodies to determine whether they could interact with antigen-derived peptides that could cause interference in the main assay. Additional Lrig-1 peptide microarray copies were then incubated with the H6 monoclonal antibody of Production Example 8 (1 μg / ml, 10 μg / ml, and 100 μg / ml) in incubation buffer and then stained with secondary and control antibodies. Readings were performed using the LI-COR Odyssey imaging system at a scanning intensity of 7 / 7 (red / green). Quantification of spot intensities and peptide annotation were based on 16-bit grayscale TIFF files. Microarray image analysis was performed using The results are in Figures 13 to 15 The analyzed epitope sequences are shown in Table 8 below.

[0192] [Table 8]

[0193]

[0194]

[0195] 3. Experimental Results

[0196] like Figure 13-15 As shown, it was discovered that a monoclonal antibody (H6) that specifically binds to the Lrig-1 protein present on regulatory T cells and is therefore effective in treating cancer specifically binds to and functions at an epitope in the Lrig-1 protein represented by any of the amino acid sequences of SEQ ID NOs: 18-29. Furthermore, it was discovered that the epitopes represented by SEQ ID NOs: 18 and 20 share a common leucine-rich repeat, and specifically, these epitopes contain a consensus sequence, which can be represented by the amino acid sequence of Formula 1 of the present invention, located at the fifth amino acid position from the N-terminus.

[0197] Although the present invention has been described in detail above, the scope of the present invention is not limited thereto. It will be apparent to those skilled in the art that various modifications and changes can be made without departing from the technical spirit of the present invention described in the claims.

[0198] Industrial availability

[0199] The present invention relates to an epitope of a leucine-rich and immunoglobulin-like domain 1 (Lrig-1) protein, which is an antigen present on the surface of regulatory T cells, and an antibody or antigen-binding fragment that specifically binds thereto. <110> Good T Cell Co., Ltd. <120> Epitopes of regulatory T cell surface antigens and antibodies specifically binding thereto <130> PDPB187156k01 <150> KR 10-2018-0053052 <151> 2018-05-09 <160> 45 <170> KoPatentIn 3.0 <210> 1 <211> 1093 <212> PRT <213> Homo sapiens <400> 1 Met Ala Arg Pro Val Arg Gly Gly Leu Gly Ala Pro Arg Arg Ser Pro 1 5 10 15 Cys Leu Leu Leu Leu Trp Leu Val Leu Val Arg Leu Glu Pro Val Thr 20 25 30 Ala Ala Ala Gly Pro Arg Ala Pro Cys Ala Ala Ala Cys Thr Cys Ala 35 40 45 Gly Asp Ser Leu Asp Cys Gly Gly Arg Gly Leu Ala Ala Leu Pro Gly 50 55 60 Asp Leu Pro Ser Trp Thr Arg Ser Leu Asn Leu Ser Tyr Asn Lys Leu 65 70 75 80 Ser Glu Ile Asp Pro Ala Gly Phe Glu Asp Leu Pro Asn Leu Gln Glu 85 90 95 Val Asn Leu Ser Tyr Asn Lys Leu Ser Glu Ile Asp Pro Ala Gly Phe 100 105 110 Glu Asp Leu Pro Asn Leu Gln Glu Val Tyr Leu Asn Asn Asn Glu Leu 115 120 125 Thr Ala Val Pro Ser Leu Gly Ala Ala Ser Ser His Val Val Ser Leu 130 135 140 Phe Leu Gln His Asn Lys Ile Arg Ser Val Glu Gly Ser Gln Leu Lys 145 150 155 160 Ala Tyr Leu Ser Leu Glu Val Leu Asp Leu Ser Leu Asn Asn Ile Thr 165 170 175 Glu Val Arg Asn Thr Cys Phe Pro His Gly Pro Pro Ile Lys Glu Leu 180 185 190 Asn Leu Ala Gly Asn Arg Ile Gly Thr Leu Glu Leu Gly Ala Phe Asp 195 200 205 Gly Leu Ser Arg Ser Leu Leu Thr Leu Arg Leu Ser Lys Asn Arg Ile 210 215 220 Thr Gln Leu Pro Val Arg Ala Phe Lys Leu Pro Arg Leu Thr Gln Leu 225 230 235 240 Asp Leu Asn Arg Asn Arg Ile Arg Leu Ile Glu Gly Leu Thr Phe Gln 245 250 255 Gly Leu Asn Ser Leu Glu Val Leu Lys Leu Gln Arg Asn Asn Ile Ser 260 265 270 Lys Leu Thr Asp Gly Ala Phe Trp Gly Leu Ser Lys Met His Val Leu 275 280 285 His Leu Glu Tyr Asn Ser Leu Val Glu Val Asn Ser Gly Ser Leu Tyr 290 295 300 Gly Leu Thr Ala Leu His Gln Leu His Leu Ser Asn Asn Ser Ile Ala 305 310 315 320 Arg Ile His Arg Lys Gly Trp Ser Phe Cys Gln Lys Leu His Glu Leu 325 330 335 Val Leu Ser Phe Asn Asn Leu Thr Arg Leu Asp Glu Glu Ser Leu Ala 340 345 350 Glu Leu Ser Ser Leu Ser Val Leu Arg Leu Ser His Asn Ser Ile Ser 355 360 365 His Ile Ala Glu Gly Ala Phe Lys Gly Leu Arg Ser Leu Arg Val Leu 370 375 380 Asp Leu Asp His Asn Glu Ile Ser Gly Thr Ile Glu Asp Thr Ser Gly 385 390 395 400 Ala Phe Ser Gly Leu Asp Ser Leu Ser Lys Leu Asn Leu Gly Gly Asn 405 410 415 Ala Ile Arg Ser Val Gln Phe Asp Ala Phe Val Lys Met Lys Asn Leu 420 425 430 Lys Glu Leu His Ile Ser Ser Asp Ser Phe Leu Cys Asp Cys Gln Leu 435 440 445 Lys Trp Leu Pro Pro Trp Leu Ile Gly Arg Met Leu Gln Ala Phe Val 450 455 460 Thr Ala Thr Cys Ala His Pro Glu Ser Leu Lys Gly Gln Ser Ile Phe 465 470 475 480 Ser Val Pro Pro Glu Ser Phe Val Cys Asp Asp Phe Leu Lys Pro Gln 485 490 495 Ile Ile Thr Gln Pro Glu Thr Thr Met Ala Met Val Gly Lys Asp Ile 500 505 510 Arg Phe Thr Cys Ser Ala Ala Ser Ser Ser Ser Ser Pro Met Thr Phe 515 520 525 Ala Trp Lys Lys Asp Asn Glu Val Leu Thr Asn Ala Asp Met Glu Asn 530 535 540 Phe Val His Val His Ala Gln Asp Gly Glu Val Met Glu Tyr Thr Thr 545 550 555 560 Ile Leu His Leu Arg Gln Val Thr Phe Gly His Glu Gly Arg Tyr Gln 565 570 575 Cys Val Ile Thr Asn His Phe Gly Ser Thr Tyr Ser His Lys Ala Arg 580 585 590 Leu Thr Val Asn Val Leu Pro Ser Phe Thr Lys Thr Pro His Asp Ile 595 600 605 Thr Ile Arg Thr Thr Thr Val Ala Arg Leu Glu Cys Ala Ala Thr Gly 610 615 620 His Pro Asn Pro Gln Ile Ala Trp Gln Lys Asp Gly Gly Thr Asp Phe 625 630 635 640 Pro Ala Ala Arg Glu Arg Arg Met His Val Met Pro Asp Asp Asp Val 645 650 655 Phe Phe Ile Thr Asp Val Lys Ile Asp Asp Ala Gly Val Tyr Ser Cys 660 665 670 Thr Ala Gln Asn Ser Ala Gly Ser Ile Ser Ala Asn Ala Thr Leu Thr 675 680 685 Val Leu Glu Thr Pro Ser Leu Val Val Pro Leu Glu Asp Arg Val Val 690 695 700 Ser Val Gly Glu Thr Val Ala Leu Gln Cys Lys Ala Thr Gly Asn Pro 705 710 715 720 Pro Pro Arg Ile Thr Trp Phe Lys Gly Asp Arg Pro Leu Ser Leu Thr 725 730 735 Glu Arg His His Leu Thr Pro Asp Asn Gln Leu Leu Val Val Gln Asn 740 745 750 Val Val Ala Glu Asp Ala Gly Arg Tyr Thr Cys Glu Met Ser Asn Thr 755 760 765 Leu Gly Thr Glu Arg Ala His Ser Gln Leu Ser Val Leu Pro Ala Ala 770 775 780 Gly Cys Arg Lys Asp Gly Thr Thr Val Gly Ile Phe Thr Ile Ala Val 785 790 795 800 Val Ser Ser Ile Val Leu Thr Ser Leu Val Trp Val Cys Ile Ile Tyr 805 810 815 Gln Thr Arg Lys Lys Ser Glu Glu Tyr Ser Val Thr Asn Thr Asp Glu 820 825 830 Thr Val Val Pro Pro Asp Val Pro Ser Tyr Leu Ser Ser Gln Gly Thr 835 840 845 Leu Ser Asp Arg Gln Glu Thr Val Val Arg Thr Glu Gly Gly Pro Gln 850 855 860 Ala Asn Gly His Ile Glu Ser Asn Gly Val Cys Pro Arg Asp Ala Ser 865 870 875 880 His Phe Pro Glu Pro Asp Thr His Ser Val Ala Cys Arg Gln Pro Lys 885 890 895 Leu Cys Ala Gly Ser Ala Tyr His Lys Glu Pro Trp Lys Ala Met Glu 900 905 910 Lys Ala Glu Gly Thr Pro Gly Pro His Lys Met Glu His Gly Gly Arg 915 920 925 Val Val Cys Ser Asp Cys Asn Thr Glu Val Asp Cys Tyr Ser Arg Gly 930 935 940 Gln Ala Phe His Pro Gln Pro Val Ser Arg Asp Ser Ala Gln Pro Ser 945 950 955 960 Ala Pro Asn Gly Pro Glu Pro Gly Gly Ser Asp Gln Glu His Ser Pro 965 970 975 His His Gln Cys Ser Arg Thr Ala Ala Gly Ser Cys Pro Glu Cys Gln 980 985 990 Gly Ser Leu Tyr Pro Ser Asn His Asp Arg Met Leu Thr Ala Val Lys 995 1000 1005 Lys Lys Pro Met Ala Ser Leu Asp Gly Lys Gly Asp Ser Ser Trp Thr 1010 1015 1020 Leu Ala Arg Leu Tyr His Pro Asp Ser Thr Glu Leu Gln Pro Ala Ser 1025 1030 1035 1040 Ser Leu Thr Ser Gly Ser Pro Glu Arg Ala Glu Ala Gln Tyr Leu Leu 1045 1050 1055 Val Ser Asn Gly His Leu Pro Lys Ala Cys Asp Ala Ser Pro Glu Ser 1060 1065 1070 Thr Pro Leu Thr Gly Gln Leu Pro Gly Lys Gln Arg Val Pro Leu Leu 1075 1080 1085 Leu Ala Pro Lys Ser 1090 <210> 2 <211> 3282 <212> DNA <213> Homo sapiens <400> 2 atggcgcggc cggtccgggg agggctcggg gccccgcgcc gctcgccttg ccttctcctt 60 ctctggctgc ttttgcttcg gctggagccg gtgaccgccg cggccggccc gcgggcgccc 120 tgcgcggccg cctgcacttg cgctggggac tcgctggact gcggtgggcg cgggctggct 180 gcgttgcccg gggacctgcc ctcctggacg cggagcctaa acctgagtta caacaaactc 240 tctgagattg accctgctgg ttttgaggac ttgccgaacc tacaggaagt gtacctcaat 300 aataatgagt tgacagcggt accatccctg ggcgctgctt catcacatgt cgtctctctc 360 tttctgcagc acaacaagat tcgcagcgtg gaggggagcc agctgaaggc ctacctttcc 420 ttagaagtgt tagatctgag tttgaacaac atcacggaag tgcggaacac ctgctttcca 480 cacggaccgc ctataaagga gctcaacctg gcaggcaatc ggattggcac cctggagttg 540 ggagcatttg atggtctgtc acggtcgctg ctaactcttc gcctgagcaa aaacaggatc 600 acccagcttc ctgtaagagc attcaagcta cccaggctga cacaactgga cctcaatcgg 660 aacaggattc ggctgataga gggcctcacc ttccaggggc tcaacagctt ggaggtgctg 720 aagcttcagc gaaacaacat cagcaaactg acagatgggg ccttctgggg actgtccaag 780 atgcatgtgc tgcacctgga gtacaacagc ctggtagaag tgaacagcgg ctcgctctac 840 ggcctcacgg ccctgcatca gctccacctc agcaacaatt ccatcgctcg cattcaccgc 900 aagggctgga gcttctgcca gaagctgcat gagttggtcc tgtccttcaa caacctgaca 960 cggctggacg aggagagcct ggccgagctg agcagcctga gtgtcctgcg tctcagccac 1020 aattccatca gccacattgc ggagggtgcc ttcaagggac tcaggagcct gcgagtcttg 1080 gatctggacc ataacgagat ttcgggcaca atagaggaca cgagcggcgc cttctcaggg 1140 ctcgacagcc tcagcaagct gactctgttt ggaaacaaga tcaagtctgt ggctaagaga 1200 gcattctcgg ggctggaagg cctggagcac ctgaaccttg gaggaatgc gatcagatct 1260 gtccagtttg atgcctttgt gaagatgaag aatcttaaag agctccatat cagcagcgac 1320 agcttcctgt gtgactgcca gctgaagtgg ctgccccgt ggctaattgg caggatgctg 1380 caggcctttg tgacagccac ctgtgcccac ccagaatcac tgaagggtca gagcattttc 1440 tctgtgccac cagagagttt cgtgtgcgat gacttcctga agccacagat catcacccag ccagaacca ccatggctat ggtgggcaag gacatccggt ttacatgctc agcagccagc agcagcagct cccccatgac ctttgcctgg aagaagaca atgaagtcct gaccatgca gacatggaga actttgtcca cgtccacgcg gagacgggg aagtgatgga gtacaccacc atcctgcacc tccgtcaggt cactttcggg cacgagggcc gctaccaatg tgtcatcacc 1740. aaccactttg gctccaccta ttcacataag gccaggctca ccgtgaatgt gttgccatca ttcaccaaaa cgccccacga cataaccatc cggaccacca ccatggcccg cctcgaatgt gctgccacag gtcacccaaa ccctcagatt gcctggcaga aggatggagg cacggatttc cccgctgccc gtgagcgacg catgcatgtc atgccggatg acgacgtgtt tttcatcact 1980. gatgtgaaaa tagtgacgc aggggtttac agctgtactg ctcagaactc agccggttct atttcagcta atgccaccct gactgtccta gagaccccat ccttggtggt ccccttggaa gaccgtgtgg tatctgtggg agaaacagtg gccctccaat gcaaagccac ggggaaccct ccgccccgca tcacctggtt caagggggac cgcccgctga gcctcactga gcggcaccac 2220 ttgacccctg acaaccagct cctggtggtt cagaacgtgg tggcagagga tgcgggccga 2280 tatacctgtg agatgtccaa caccctgggc acggagcgag ctcacagcca gctgagcgtc 2340 ctgcccgcag caggctgcag gaaggatggg accacggtag gcatcttcac cattgctgtc 2400 gtgagcagca tcgtcctgac gtcactggtc tgggtgtgca tcatctacca gaccaggaag 2460 aagagtgaag agtacagtgt caccaacaca gatgaaaccg tcgtgccacc agatgttcca 2520 agctacctct cttctcaggg gaccctttct gaccgacaag aaaccgtggt caggaccgag 2580 ggtggccctc aggccaatgg gcacattgag agcaatggtg tgtgtccaag agatgcaagc 2640 cactttccag agcccgacac tcacagcgtt gcctgcaggc agccaaagct ctgtgctggg 2700 tctgcgtatc acaaagagcc gtggaaagcg atggagaaag ctgaagggac acctgggcca 2760 cataagatgg aacacggtgg ccgggtcgta tgcagtgact gcaacaccga agtggactgt 2820 tactccaggg gacaagcctt ccacccccag cctgtgtcca gagacagcgc acagccaagt 2880 gcgccaaatg gcccggagcc gggtgggagt gaccaagagc attctccaca tcaccagtgc 2940 agcaggactg ccgctgggtc ctgccccgag tgccaagggt cgctctaccc cagtaaccac 3000 gatagaatgc tgacggctgt gaagaaaaag ccaatggcat ctctagatgg gaaaggggat 3060 tcttcctgga ctttagcaag gttgtatcac ccggactcca cagagctaca gcctgcatct 3120 tcattaactt caggcagtcc agagcgcgcg gaagcccagt acttgcttgt ttccaatggc 3180 cacctcccca aagcatgtga cgccagtccc gagtccacgc cactgacagg acagctcccc 3240 gggaaacaga gggtgccact gctgttggca ccaaaaagct ag 3282 <210> 3 <211> 1091 <212> PRT <213> Mouse (Mus musculus) <400> 3 Met Ala Arg Pro Gly Pro Gly Val Leu Gly Ala Pro Arg Leu Ala Pro 1 5 10 15 Arg Leu Leu Leu Trp Leu Leu Leu Leu Leu Leu Gln Trp Pro Glu Ser 20 25 30 Ala Gly Ala Gln Ala Gly Pro Arg Ala Pro Cys Ala Ala Ala Cys Thr 35 40 45 Cys Ala Gly Asp Ser Leu Asp Cys Ser Gly Arg Gly Leu Ala Thr Leu 50 55 60 Pro Arg Asp Leu Pro Ser Trp Thr Arg Ser Leu Asn Leu Ser Tyr Asn 65 70 75 80 Arg Leu Ser Glu Ile Asp Ser Ala Ala Phe Glu Asp Leu Thr Asn Leu 85 90 95 Gln Glu Val Tyr Leu Asn Ser Asn Glu Leu Thr Ala Ile Pro Ser Leu 100 105 110 Gly Thr Ala Ser Ile Gly Val Val Ser Leu Phe Leu Gln His Asn Lys 115 120 125 Ile Leu Ser Val Asp Gly Ser Gln Leu Lys Ser Tyr Leu Ser Leu Glu 130 135 140 Val Leu Asp Leu Ser Ser Asn Asn Ile Thr Glu Ile Arg Ser Ser Cys 145 150 155 160 Phe Pro Asn Gly Leu Arg Ile Arg Glu Leu Asn Leu Ala Ser Asn Arg 165 170 175 Ile Ser Ile Leu Glu Ser Gly Ala Phe Asp Gly Leu Ser Arg Ser Leu 180 185 190 Leu Thr Leu Arg Leu Ser Lys Asn Arg Ile Thr Gln Leu Pro Val Lys 195 200 205 Ala Phe Lys Leu Pro Arg Leu Thr Gln Leu Asp Leu Asn Arg Asn Arg 210 215 220 Ile Arg Leu Ile Glu Gly Leu Thr Phe Gln Gly Leu Asp Ser Leu Glu 225 230 235 240 Val Leu Arg Leu Gln Arg Asn Asn Ile Ser Arg Leu Thr Asp Gly Ala 245 250 255 Phe Trp Gly Leu Ser Lys Met His Val Leu His Leu Glu Tyr Asn Ser 260 265 270 Leu Val Glu Val Asn Ser Gly Ser Leu Tyr Gly Leu Thr Ala Leu His 275 280 285 Gln Leu His Leu Ser Asn Asn Ser Ile Ser Arg Ile Gln Arg Asp Gly 290 295 300 Trp Ser Phe Cys Gln Lys Leu His Glu Leu Ile Leu Ser Phe Asn Asn 305 310 315 320 Leu Thr Arg Leu Asp Glu Glu Ser Leu Ala Glu Leu Ser Ser Leu Ser 325 330 335 Ile Leu Arg Leu Ser His Asn Ala Ile Ser His Ile Ala Glu Gly Ala 340 345 350 Phe Lys Gly Leu Lys Ser Leu Arg Val Leu Asp Leu Asp His Asn Glu 355 360 365 Ile Ser Gly Thr Ile Glu Asp Thr Ser Gly Ala Phe Thr Gly Leu Asp 370 375 380 Asn Leu Ser Lys Leu Thr Leu Phe Gly Asn Lys Ile Lys Ser Val Ala 385 390 395 400 Lys Arg Ala Phe Ser Gly Leu Glu Ser Leu Glu His Leu Asn Leu Gly 405 410 415 Glu Asn Ala Ile Arg Ser Val Gln Phe Asp Ala Phe Ala Lys Met Lys 420 425 430 Asn Leu Lys Glu Leu Tyr Ile Ser Ser Glu Ser Phe Leu Cys Asp Cys 435 440 445 Gln Leu Lys Trp Leu Pro Pro Trp Leu Met Gly Arg Met Leu Gln Ala 450 455 460 Phe Val Thr Ala Thr Cys Ala His Pro Glu Ser Leu Lys Gly Gln Ser 465 470 475 480 Ile Phe Ser Val Leu Pro Asp Ser Phe Val Cys Asp Asp Phe Pro Lys 485 490 495 Pro Gln Ile Ile Thr Gln Pro Glu Thr Thr Met Ala Val Val Gly Lys 500 505 510 Asp Ile Arg Phe Thr Cys Ser Ala Ala Ser Ser Ser Ser Ser Pro Met 515 520 525 Thr Phe Ala Trp Lys Lys Asp Asn Glu Val Leu Ala Asn Ala Asp Met 530 535 540 Glu Asn Phe Ala His Val Arg Ala Gln Asp Gly Glu Val Met Glu Tyr 545 550 555 560 Thr Thr Ile Leu His Leu Arg His Val Thr Phe Gly His Glu Gly Arg 565 570 575 Tyr Gln Cys Ile Ile Thr Asn His Phe Gly Ser Thr Tyr Ser His Lys 580 585 590 Ala Arg Leu Thr Val Asn Val Leu Pro Ser Phe Thr Lys Ile Pro His 595 600 605 Asp Ile Ala Ile Arg Thr Gly Thr Thr Ala Arg Leu Glu Cys Ala Ala 610 615 620 Thr Gly His Pro Asn Pro Gln Ile Ala Trp Gln Lys Asp Gly Gly Thr 625 630 635 640 Asp Phe Pro Ala Ala Arg Glu Arg Arg Met His Val Met Pro Asp Asp 645 650 655 Asp Val Phe Phe Ile Thr Asp Val Lys Ile Asp Asp Met Gly Val Tyr 660 665 670 Ser Cys Thr Ala Gln Asn Ser Ala Gly Ser Val Ser Ala Asn Ala Thr 675 680 685 Leu Thr Val Leu Glu Thr Pro Ser Leu Ala Val Pro Leu Glu Asp Arg 690 695 700 Val Val Thr Val Gly Glu Thr Val Ala Phe Gln Cys Lys Ala Thr Gly 705 710 715 720 Ser Pro Thr Pro Arg Ile Thr Trp Leu Lys Gly Gly Arg Pro Leu Ser 725 730 735 Leu Thr Glu Arg His His Phe Thr Pro Gly Asn Gln Leu Leu Val Val 740 745 750 Gln Asn Val Met Ile Asp Asp Ala Gly Arg Tyr Thr Cys Glu Met Ser 755 760 765 Asn Pro Leu Gly Thr Glu Arg Ala His Ser Gln Leu Ser Ile Leu Pro 770 775 780 Thr Pro Gly Cys Arg Lys Asp Gly Thr Thr Val Gly Ile Phe Thr Ile 785 790 795 800 Ala Val Val Cys Ser Ile Val Leu Thr Ser Leu Val Trp Val Cys Ile 805 810 815 Ile Tyr Gln Thr Arg Lys Lys Ser Glu Glu Tyr Ser Val Thr Asn Thr 820 825 830 Asp Glu Thr Ile Val Pro Pro Asp Val Pro Ser Tyr Leu Ser Ser Gln 835 840 845 Gly Thr Leu Ser Asp Arg Gln Glu Thr Val Val Arg Thr Glu Gly Gly 850 855 860 His Gln Ala Asn Gly His Ile Glu Ser Asn Gly Val Cys Leu Arg Asp 865 870 875 880 Pro Ser Leu Phe Pro Glu Val Asp Ile His Ser Thr Thr Cys Arg Gln 885 890 895 Pro Lys Leu Cys Val Gly Tyr Thr Arg Glu Pro Trp Lys Val Thr Glu 900 905 910 Lys Ala Asp Arg Thr Ala Ala Pro His Thr Thr Ala His Ser Gly Ser 915 920 925 Ala Val Cys Ser Asp Cys Ser Thr Asp Thr Ala Tyr His Pro Gln Pro 930 935 940 Val Pro Arg Asp Ser Gly Gln Pro Gly Thr Ala Ser Ser Gln Glu Leu 945 950 955 960 Arg Gln His Asp Arg Glu Tyr Ser Pro His His Pro Tyr Ser Gly Thr 965 970 975 Ala Asp Gly Ser His Thr Leu Ser Gly Gly Ser Leu Tyr Pro Ser Asn 980 985 990 His Asp Arg Ile Leu Pro Ser Leu Lys Asn Lys Ala Ala Ser Ala Asp 995 1000 1005 Gly Asn Gly Asp Ser Ser Trp Thr Leu Ala Lys Leu His Glu Ala Asp 1010 1015 1020 Cys Ile Asp Leu Lys Pro Ser Pro Thr Leu Ala Ser Gly Ser Pro Glu 1025 1030 1035 1040 Leu Met Glu Asp Ala Ile Ser Thr Glu Ala Gln His Leu Leu Val Ser 1045 1050 1055 Asn Gly His Leu Pro Lys Ala Cys Asp Ser Ser Pro Glu Ser Val Pro 1060 1065 1070 Leu Lys Gly Gln Ile Thr Gly Lys Arg Arg Gly Pro Leu Leu Leu Ala 1075 1080 1085 Pro Arg Serum 1090 <210> 4 <211> 6 <212> PRT <213> Artificial Sequence <400> 4 Leo Asn Leo Ser Tyr Asn 1 5 <210> 5 <211> 6 <212> PRT <213> Artificial Sequence <400> 5 Leu Asp Leu Asn Arg Asn 1 5 <210> 6 <211> 6 <212> PRT <213> Artificial Sequence <400> 6 Leu Phe Leu Gln His Asn 1 5 <210> 7 <211> 6 <212> PRT <213> Artificial Sequence <400> 7 Leu Asn Leu Ala Gly Asn 1 5 <210> 8 <211> 6 <212> PRT <213> Artificial Sequence <400> 8 Leu Asp Leu Ser Leu Asn 1 5 <210> 9 <211> 6 <212> PRT <213> Artificial Sequence <400> 9 Leu Arg Leu Ser Lys Asn 1 5 <210> 10 <211> 6 <212> PRT <213> Artificial Sequence <400> 10 Leu Lys Leu Gln Arg Asn 1 5 <210> 11 <211> 6 <212> PRT <213> Artificial Sequence <400> 11 Leu His Leu Glu Tyr Asn 1 5 <210> 12 <211> 6 <212> PRT <213> Artificial Sequence <400> 12 Leu His Leu Ser Asn Asn 1 5 <210> 13 <211> 6 <212> PRT <213> Artificial Sequence <400> 13 Leu Val Leu Ser Phe Asn 1 5 <210> 14 <211> 6 <212> PRT <213> Artificial Sequence <400> 14 Leu Arg Leu Ser His Asn 1 5 <210> 15 <211> 6 <212> PRT <213> Artificial Sequence <400> 15 Leu Asp Leu Asp His Asn 1 5 <210> 16 <211> 6 <212> PRT <213> Artificial Sequence <400> 16 Leu Thr Leu Phe Gly Asn 1 5 <210> 17 <211> 6 <212> PRT <213> Artificial Sequence <400> 17 Leu Asn Leu Gly Gly Asn 1 5 <210> 18 <211> 12 <212> PRT <213> Artificial Sequence <400> 18 Trp Thr Arg Ser Leu Asn Leu Ser Tyr Asn Lys Leu 1 5 10 <210> 19 <211> 14 <212> PRT <213> Artificial Sequence <400> 19 Thr Glu Val Arg Asn Thr Cys Phe Pro His Gly Pro Pro Ile 1 5 10 <210> 20 <211> 13 <212> PRT <213> Artificial Sequence <400> 20 Arg Leu Thr Gln Leu Asp Leu Asn Arg Asn Arg Ile Arg 1 5 10 <210> twenty one <211> 15 <212> PRT <213> Artificial Sequence <400> twenty one Asp Leu Asn Arg Asn Arg Ile Arg Leu Ile Glu Gly Leu Thr Phe 1 5 10 15 <210> twenty two <211> 11 <212> PRT <213> Artificial Sequence <400> twenty two Asn Ser Ile Ala Arg Ile His Arg Lys Gly Trp 1 5 10 <210> twenty three <211> 14 <212> PRT <213> Artificial Sequence <400> twenty three Trp Leu Pro Pro Trp Leu Ile Gly Arg Met Leu Gln Ala Phe 1 5 10 <210> twenty four <211> 11 <212> PRT <213> Artificial Sequence <400> twenty four Arg Gln Val Thr Phe Gly His Glu Gly Arg Tyr 1 5 10 <210> 25 <211> 15 <212> PRT <213> Artificial Sequence <400> 25 Phe Gly His Glu Gly Arg Tyr Gln Cys Val Ile Thr Asn His Phe 1 5 10 15 <210> 26 <211> 15 <212> PRT <213> Artificial Sequence <400> 26 Arg Leu Thr Val Asn Val Leu Pro Ser Phe Thr Lys Thr Pro His 1 5 10 15 <210> 27 <211> 13 <212> PRT <213> Artificial Sequence <400> 27 Arg Arg Met His Val Met Pro Asp Asp Asp Val Phe Phe 1 5 10 <210> 28 <211> 15 <212> PRT <213> Artificial Sequence <400> 28 Phe Phe Ile Thr Asp Val Lys Ile Asp Asp Ala Gly Val Tyr Ser 1 5 10 15 <210> 29 <211> 13 <212> PRT <213> Artificial Sequence <400> 29 Lys Gly Asp Arg Pro Leu Ser Leu Thr Glu Arg His His 1 5 10 <210> 30 <211> 449 <212> PRT <213> Artificial Sequence <400> 30 Glu Val Gln Leu Leu 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 Gly Tyr 20 25 30 Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Leu Ile Tyr Pro Asp Ser Gly Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Ala Gly Leu Ser Trp Ala Gly Ala Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Thr Thr Ala Pro Ser Val Tyr 115 120 125 Pro Leu Ala Pro Val Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu 130 135 140 Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Leu Thr Trp 145 150 155 160 Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Thr Ser Ser 180 185 190 Thr Trp Pro Ser Gln Ser Ile Thr Cys Asn Val Ala His Pro Ala Ser 195 200 205 Ser Thr Lys Val Asp Lys Lys Ile Glu Pro Arg Gly Pro Thr Ile Lys 210 215 220 Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro Asn Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Ile Phe Pro Pro Lys Ile Lys Asp Val Leu Met Ile Ser 245 250 255 Leu Ser Pro Ile Val Thr Cys Val Val Val Asp Val Ser Glu Asp Asp 260 265 270 Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val Glu Val His Thr 275 280 285 Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Leu Arg Val 290 295 300 Val Ser Ala Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly Lys Glu 305 310 315 320 Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ala Pro Ile Glu Arg 325 330 335 Thr Ile Ser Lys Pro Lys Gly Ser Val Arg Ala Pro Gln Val Tyr Val 340 345 350 Leu Pro Pro Pro Glu Glu Glu Met Thr Lys Lys Gln Val Thr Leu Thr 355 360 365 Cys Met Val Thr Asp Phe Met Pro Glu Asp Ile Tyr Val Glu Trp Thr 370 375 380 Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys Asn Thr Glu Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser Lys Leu Arg Val Glu Lys 405 410 415 Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser Cys Ser Val Val His Glu 420 425 430 Gly Leu His Asn His His Thr Thr Lys Ser Phe Ser Arg Thr Pro Gly 435 440 445 Lys <210> 31 <211> 217 <212> PRT <213> Artificial Sequence <400> 31 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ser Asp Ser His Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Ser Trp Asp Tyr Ser Leu 85 90 95 Ser Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn 145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His 180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile 195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 32 <211> 449 <212> PRT <213> Artificial Sequence <400> 32 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Pro Gly Asp Ser Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Leu Tyr Ser Asn Pro Asn Glu Pro Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Thr Thr Ala Pro Ser Val Tyr 115 120 125 Pro Leu Ala Pro Val Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu 130 135 140 Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Leu Thr Trp 145 150 155 160 Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Thr Ser Ser 180 185 190 Thr Trp Pro Ser Gln Ser Ile Thr Cys Asn Val Ala His Pro Ala Ser 195 200 205 Ser Thr Lys Val Asp Lys Lys Ile Glu Pro Arg Gly Pro Thr Ile Lys 210 215 220 Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro Asn Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Ile Phe Pro Pro Lys Ile Lys Asp Val Leu Met Ile Ser 245 250 255 Leu Ser Pro Ile Val Thr Cys Val Val Val Asp Val Ser Glu Asp Asp 260 265 270 Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val Glu Val His Thr 275 280 285 Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Leu Arg Val 290 295 300 Val Ser Ala Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly Lys Glu 305 310 315 320 Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ala Pro Ile Glu Arg 325 330 335 Thr Ile Ser Lys Pro Lys Gly Ser Val Arg Ala Pro Gln Val Tyr Val 340 345 350 Leu Pro Pro Pro Glu Glu Glu Met Thr Lys Lys Gln Val Thr Leu Thr 355 360 365 Cys Met Val Thr Asp Phe Met Pro Glu Asp Ile Tyr Val Glu Trp Thr 370 375 380 Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys Asn Thr Glu Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser Lys Leu Arg Val Glu Lys 405 410 415 Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser Cys Ser Val Val His Glu 420 425 430 Gly Leu His Asn His His Thr Thr Lys Ser Phe Ser Arg Thr Pro Gly 435 440 445 Lys <210> 33 <211> 217 <212> PRT <213> Artificial Sequence <400> 33 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Asp Asp Ser Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Tyr Ser Leu 85 90 95 Asn Gly Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn 145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His 180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile 195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 34 <211> 455 <212> PRT <213> Artificial Sequence <400> 34 Glu Val Gln Leu Leu 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 Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Pro Asp Gly Ser Asn Ile Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Val Gly Leu Arg Cys Arg Tyr Glu Ala Cys Ser Tyr Ala Tyr 100 105 110 Gly Met Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Thr 115 120 125 Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro Val Cys Gly Asp Thr Thr 130 135 140 Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Leu Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser 180 185 190 Val Thr Val Thr Ser Ser Thr Trp Pro Ser Gln Ser Ile Thr Cys Asn 195 200 205 Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Glu Pro 210 215 220 Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro 225 230 235 240 Asn Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys 245 250 255 Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val Val 260 265 270 Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn 275 280 285 Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr 290 295 300 Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp 305 310 315 320 Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu 325 330 335 Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val Arg 340 345 350 Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys 355 360 365 Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu Asp 370 375 380 Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys 385 390 395 400 Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser 405 410 415 Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser 420 425 430 Cys Ser Val Val His Glu Gly Leu His Asn His His Thr Thr Lys Ser 435 440 445 Phe Ser Arg Thr Pro Gly Lys 450 455 <210> 35 <211> 217 <212> PRT <213> Artificial Sequence < Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ser Asp Ser His Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Trp Asp Ser Ser Leu 85 90 95 Asn Gly Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn 145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His 180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile 195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 36 <211> 449 <212> PRT <213> Artificial Sequence <400> 36 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Pro Ser Ser Gly Ser Ile Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Asp Leu Asp Ala Phe Trp Arg Pro Ser Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Thr Thr Ala Pro Ser Val Tyr 115 120 125 Pro Leu Ala Pro Val Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu 130 135 140 Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Leu Thr Trp 145 150 155 160 Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Thr Ser Ser 180 185 190 Thr Trp Pro Ser Gln Ser Ile Thr Cys Asn Val Ala His Pro Ala Ser 195 200 205 Ser Thr Lys Val Asp Lys Lys Ile Glu Pro Arg Gly Pro Thr Ile Lys 210 215 220 Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro Asn Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Ile Phe Pro Pro Lys Ile Lys Asp Val Leu Met Ile Ser 245 250 255 Leu Ser Pro Ile Val Thr Cys Val Val Val Asp Val Ser Glu Asp Asp 260 265 270 Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val Glu Val His Thr 275 280 285 Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Leu Arg Val 290 295 300 Val Ser Ala Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly Lys Glu 305 310 315 320 Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ala Pro Ile Glu Arg 325 330 335 Thr Ile Ser Lys Pro Lys Gly Ser Val Arg Ala Pro Gln Val Tyr Val 340 345 350 Leu Pro Pro Pro Glu Glu Glu Met Thr Lys Lys Gln Val Thr Leu Thr 355 360 365 Cys Met Val Thr Asp Phe Met Pro Glu Asp Ile Tyr Val Glu Trp Thr 370 375 380 Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys Asn Thr Glu Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser Lys Leu Arg Val Glu Lys 405 410 415 Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser Cys Ser Val Val His Glu 420 425 430 Gly Leu His Asn His His Thr Thr Lys Ser Phe Ser Arg Thr Pro Gly 435 440 445 Lys <210> 37 <211> 217 <212> PRT <213> Artificial Sequence <400> 37 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Asn Asn<l 20 25 30 Asn Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ser Asp Ser His Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Ser Trp Asp Asp Ser Leu 85 90 95 Ser Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn IS145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 1'75 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His [[ID='26]]180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile 195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 38 <211> 455 <212> PRT <213> Artificial Sequence <400> 38 It should be noted that there seems to be a small error in the original text where "IS145" in line 18 should probably be "145". The translation has been done as accurately as possible based on the provided text.Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Asp Met Ser Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Trp Ile Ser His Gly Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Leu Gly Leu Cys Lys Thr Gly Leu Cys Tyr Tyr Tyr Asp 100 105 110 Ala Met Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Thr 115 120 125 Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro Val Cys Gly Asp Thr Thr 130 135 140 Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Leu Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser 180 185 190 Val Thr Val Thr Ser Ser Thr Trp Pro Ser Gln Ser Ile Thr Cys Asn 195 200 205 Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Glu Pro 210 215 220 Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro 225 230 235 240 Asn Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys 245 250 255 Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val Val 260 265 270 Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn 275 280 285 Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr 290 295 300 Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp 305 310 315 320 Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu 325 330 335 Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val Arg 340 345 350 Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys 355 360 365 Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu Asp 370 375 380 Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys 385 390 ۳۹۵ 400 Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser 405 410 415 Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser 420 425 430 Cys Ser Val Val His Glu Gly Leu His Asn His His Thr Thr Lys Ser 435 440 445 Phe Ser Arg Thr Pro Gly Lys 450 455 <210> 39 <211> 217 <212> PRT <213> Artificial Sequence <400> 39 It should be noted that in the original text, "۳۹۵" in line 19 seems to be a typo. I translated it as "395" in the English version. If this is not what you intended, please let me know. Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Asn Asn 20 25 30 Ser Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ala Asp Asn Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Ser Ser Leu 85 90 95 Ser Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn 145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His 180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile 195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 40 <211> 444 <212> PRT <213> Artificial Sequence <400> 40 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser His Asp Ser Gly Ser Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg His Trp Thr Thr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Thr Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro Val 115 120 125 Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys 130 135 140 Gly Tyr Phe Pro Glu Pro Val Thr Leu Thr Trp Asn Ser Gly Ser Leu 145 150 155 160 Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr 165 170 175 Thr Leu Ser Ser Ser Val Thr Val Thr Ser Ser Thr Trp Pro Ser Gln 180 185 190 Ser Ile Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val Asp 195 200 205 Lys Lys Ile Glu Pro Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys 210 215 220 Lys Cys Pro Ala Pro Asn Leu Leu Gly Gly Pro Ser Val Phe Ile Phe 225 230 235 240 Pro Pro Lys Ile Lys Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val 245 250 255 Thr Cys Val Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile 260 265 270 Ser Trp Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr 275 280 285 His Arg Glu Asp Tyr Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro 290 295 300 Ile Gln His Gln Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val 305 310 315 320 Asn Asn Lys Asp Leu Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro 325 330 335 Lys Gly Ser Val Arg Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu 340 345 350 Glu Glu Met Thr Lys Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp 355 360 365 Phe Met Pro Glu Asp Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr 370 375 380 Glu Leu Asn Tyr Lys Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser 385 390 395 400 Tyr Phe Met Tyr Ser Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu 405 410 415 Arg Asn Ser Tyr Ser Cys Ser Val Val His Glu Gly Leu His Asn His 420 425 430 His Thr Thr Lys Ser Phe Ser Arg Thr Pro Gly Lys 435 440 <210> 41 <211> 217 <212> PRT <213> Artificial Sequence <400> 41 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Asn Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ala Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Ala Trp Asp Tyr Ser Leu 85 90 95 Ser Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn 145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His 180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile<![CDATA[ ]]>195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 42 <211> 455 <212> PRT <213> Artificial Sequence <400> 42 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Tyr Pro Gly Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Ile Leu Pro Cys Pro Trp Gly Arg Cys Tyr Tyr Asp Tyr 100 105 110 Ala Met Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Thr 115 120 125 Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro Val Cys Gly Asp Thr Thr 130 135 140 Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Leu Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser 180 185 190 Val Thr Val Thr Ser Ser Thr Trp Pro Ser Gln Ser Ile Thr Cys Asn 195 200 205 Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Glu Pro 210 215 220 Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro 225 230 235 240 Asn Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys 245 250 255 Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val Val 260 265 270 Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn 275 280 285 Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr 290 295 300 Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp 305 310 315 320 Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu 325 330 335 Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val Arg 340 345 350 Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys 355 360 365 Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu Asp 370 375 380 Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys 385 390 395 400 Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser 405 410 415 Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser 420 425 430 Cys Ser Val Val His Glu Gly Leu His Asn His His Thr Thr Lys Ser 435 440 445 Phe Ser Arg Thr Pro Gly Lys 450 455 <210> 43 <211> 217 <212> PRT <213> Artificial Sequence <400> 43 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Asp Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30 Thr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ala Asp Asn Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Tyr Ser Leu 85 90 95 Ser Gly Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn 145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His 180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile 195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 44 <211> 455 <212> PRT <213> Artificial Sequence <400> 44 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Ser His Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Ile Ser Asn Cys His Leu Gly Val Cys Tyr Tyr Ser Asn 100 105 110 Gly Met Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Thr 115 120 125 Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro Val Cys Gly Asp Thr Thr 130 135 140 Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Leu Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser 180 185 190 Val Thr Val Thr Ser Ser Thr Trp Pro Ser Gln Ser Ile Thr Cys Asn 195 200 205 Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Glu Pro 210 215 220 Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro 225 230 235 240 Asn Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys 245 250 255 Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val Val 260 265 270 Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn 275 280 285 Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr 290 295 300 Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp 305 310 315 320 Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu 325 330 335 Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val Arg 340 345 350 Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys 355 360 365 Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu Asp 370 375 380 Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys 385 390 395 400 Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser 405 410 415 Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser 420 425 430 Cys Ser Val Val His Glu Gly Leu His Asn His His Thr Thr Lys Ser 435 440 445 Phe Ser Arg Thr Pro Gly Lys 450 455 <210> 45 <211> 217 <212> PRT <213> Artificial Sequence <400> 45 Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn 20 25 30 Asp Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ser Asp Ser Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Tyr Ser Leu 85 90 95 Ser Gly Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr 100 105 110 Val Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu 115 120 125 Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 130 135 140 Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn 145 150 155 160 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr 165 170 175 Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His 180 185 190 Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile 195 200 205 Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215

Claims

1. A peptide derived from the leucine-rich immunoglobulin-like domain 1 (Lrig-1) protein, A polypeptide consisting of any one of the amino acid sequences of SEQ ID NOs: 18 to 29.

2. A nucleic acid molecule encoding the peptide according to claim 1.

3. An expression vector, into which the nucleic acid molecule according to claim 2 is inserted. A host cell line transfected with the expression vector according to claim 3 .

5. Use of an antibody or antigen-binding fragment for preparing a pharmaceutical composition for preventing or treating cancer, the pharmaceutical composition comprising as an active ingredient the antibody or antigen-binding fragment: in, The cancer is a solid tumor; The antibody or antibody binding fragment specifically binds to a peptide derived from the leucine-rich and immunoglobulin-like domain 1 (Lrig-1) protein, which is present on the surface of regulatory T cells and consists of any one of the amino acid sequences of SEQ ID NOs: 18 to 29; The antibody or antibody binding fragment comprises: a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 44; and The light chain variable region consists of the amino acid sequence shown in SEQ ID NO:

45.

6. The use according to claim 5, characterized in that The cancer is gastric cancer, liver cancer, glioblastoma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer.

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