CN101583623A - Fusion protein comprising tumor rejection antigens NY-ESO-1 and LAGE-1 - Google Patents

Abstract

Fusion proteins comprising an antigen derived from NY-ESO-1 linked to an antigen derived from LAGE-1 are provided, which may further comprise a carrier, a fusion partner, and the like. Methods for making, formulating, and using such fusion proteins are also provided. Such proteins are useful vaccine components for inducing immune responses against a range of cells bearing cancer antigens.

Description

The fusion rotein that comprises Tumor rejection antigen NY-ESO-1 and LAGE-1

Technical field

Generally speaking, the present invention relates to comprise derived from one or both antigenic polypeptide and the construct among Tumor rejection antigen NY-ESO-1 and the LAGE-1.

Background technology

Cancer testis (CT) antigen is that it expresses a class tumor associated antigen that normally is limited to the sexual cell among testis, ovary or the trophocyte.These antigens are not expressed in adult body cell tissue usually.Referring to, people such as Simpson, Nat.Rev.Cancer, 5 (8): 615-625 (2005); Scanlan waits the people, Immunol.Reviews, 188:22-32 (2002); Scanlan waits the people, Canc.Immun., 4:1-15 (2004).

The antigenic generegulation of CT is destroyed in the cancer patients, thereby causes these antigens unconventionality expression in extensively various tumour.First kind of CT antigen MAGE-1 to be identified obtains identifying (people such as van der Bruggen, 1991 Science 13 by t cell epitope clone in early days in the nineties in 20th century; 254 (5038): 1643-7; People such as van der Bruggen, 1999 Science254:1643-1647; Traversari waits the people, 1992 Immunogenetics, 35 (3): 145-152; With U.S. Patent number 5,342,774, be incorporated herein by reference).From then on, serology cloning by expression technology (SEREX) (Sahin, Deng the people, Proc.Natl.Acad.Sci.USA, 92 (25): 11810-11813 (1995) and U.S. Patent number 5,698,396), the recombinant antigen on yeast surface is expressed (RAYS) (Mischo, Deng the people, Canc.Immun., 3:5-16 (2003)) and difference mRNA expression analysis (Gure, Deng the people, Int.J.Canc., 85 (5): 726-732 (2000)) caused about 90 kinds of antigenic evaluations of CT, and their number is expected in the following time and increases.Number of C T immunogenicity of antigens among the cancer patients makes them become the desirable target that is used to develop tumor vaccine.

NY-ESO-1。The present interested cancer testis antigen that is used for using at cancer immunotherapy is NY-ESO-1.This antigen at first in esophageal squamous cell carcinoma obtains identify (Chen in the late period nineties at New YorkBranch of the Ludwig Institute for Cancer Research by SEREX, Deng the people, PNAS USA, 94 (5): 1914-1918 (1997); With U.S. Patent number 5,804,381, be incorporated herein by reference).

Long 180 amino acid of protein N Y-ESO-1, and can be described as forming by 3 zones:

About or the about amino acid/11-70 in ■ N-terminal zone,

The ■ middle section approximately or approximately amino acid 71-134 and

About or the about amino acid/11 35-180 in ■ C-terminal zone.

Collagen sample zone comprises the about or about amino acid/11 5-73 of pact (referring to Fig. 1) in N-terminal zone.

Protein N Y-ESO-1 finds in extensively various tumour, includes but not limited in ovarian cancer, lung cancer, mammary cancer, prostate gland (prostrate), the esophageal carcinoma, bladder cancer and the melanoma.(Nicholaou T waits the people, Immunol Cell Biol.2006 Jun; 84 (3): 303-17 and Jungbluth, wait people 2001, Int.J.Canc., 92 (6): 856-860).In patient, describe at this antigenic spontaneous body fluid and cellullar immunologic response with NY-ESO-1 positive tumor, and the peptide of many HLA (human leucocyte antigen (HLA)) I and the restriction of II class has obtained identifying (Jager, Deng the people, 1998J.Exp.Med., 187 (2): 265-270; Yamaguchi waits the people, 2004 Clin.Canc.Res., 10 (3): 890-961; And Davis, wait the people, 2004Proc.Natl.Acad.Sci.USA, 101 (29): 10697-10702).Exemplary patent documentation is a U.S. Patent number 6,140,050; 6,251,603; 6,242,052; 6,274,145; 6,338,947; 6,417,165; 6,525,177; 6,605,711; 6,689,742; 6,723,832; 6,756,044; With 6,800,730, all be incorporated herein by reference.

In clinical trial, have for 3 kinds of partly overlapping NY-ESO-1 derived peptide (157-167,157-165 and 155-163) of the binding motif of HLA-A2 and in vaccine, use to be used for the treatment of 12 patients with transitivity NY-ESO-1 expressing tumor.This studies confirm that synthesizing the NY-ESO-1 peptide can use safely, and (Jager waits the people, 2000 PNAS USA, 97 (22): 12198-12203) can to produce potential favourable t cell response.

Many MHC in the protein (main histocompatibility complex) I is obtained identifying by different groups with II class epi-position, referring to for example Fig. 1.These epi-positions only are the representatives for the epi-position of protein report, and the tabulation among Fig. 1 is not detailed.In addition, report and/or at least a or multiple epi-position listed are not confirmed as yet by experiment among Fig. 1.Collagen sample zone in the N-terminal comprises at least a MHC I class epi-position that is referred to herein as A31.Middle section comprises several MHC 2 class epi-positions that are referred to herein as DR1, DR2, DR4, DR7 and DP4.This zone also comprises several MHC I class epi-positions that are referred to herein as B35, B51, Cw3 and Cw6.C-terminal is considered to comprise at least 2 kinds of II class epi-positions (DR4 and DP4) and a kind of I class epi-position (A2).

LAGE-1。Further cancer testis antigen LAGE-1 has also obtained identifying.2 kinds of LAGE-1 transcript LAGE-1a and LAGE1b have obtained describing.LAGE1b is incomplete montage, and the protein of inferring of about 210 amino-acid residues of encoding, and the LAGE-1a gene product comprises 180 amino-acid residues (people Cancer Immunol Immunother 2006:55:644-652 such as Sun).

The proteinic N-terminal of LAGE-1 and NY-ESO-1 zone is a high conservative, and is considered to have above 97% identity.Yet LAGE-1 is different from NY-ESO-1 in middle section, and described middle section only 62% is equal to.The C-terminal of NY-ESO-1 and LAGE-1a is high conservative (surpassing 97% identity).Yet the C-terminal of LAGE-1b is longer and do not guard, and be considered to LAGE-1a/NY-ESO-1 in same area have less than 50% identity.

The general information relevant with these protein can obtain from LICR website (referring to www.cancerimmunity.org/CTdatabase).

Summary of the invention

The invention provides and comprise following immunogenicity fusion rotein:

(i) NY-ESO-1 or its fragment, it is connected to

(ii) LAGE-1 or its fragment,

Wherein at least a among NY-ESO-1 and/or the LAGE-1 be brachymemma or the part brachymemma, or comprise the fragment of one or more epi-positions of NY-ESO-1 or LAGE-1.The present invention also provides and has comprised following immunogenicity fusion rotein:

(i) LAGE-1 or its fragment, it is connected to

(ii) NY-ESO-1 or its fragment,

Wherein at least a among NY-ESO-1 and/or the LAGE-1 be brachymemma or the part brachymemma, or comprise the fragment of one or more epi-positions of NY-ESO-1 or LAGE-1.Therefore, also provide its segmental polypeptide and the fusion rotein that comprises NY-ESO-1 brachymemma or the part brachymemma or comprise one or more epi-positions of NY-ESO-1.Its segmental polypeptide and fusion rotein of comprising LAGE-1 brachymemma or the part brachymemma or comprising one or more epi-positions of LAGE-1 also are provided.The composition and the method that relate to this kind fusion rotein and polypeptide also are provided.

Description of drawings

Fig. 1 is presented at many MHC (main histocompatibility complex) I and the II class epi-position of having been identified by different groups on the NY-ESO-1 protein.These epi-positions only are the representatives for the epi-position of protein report, so the tabulation among Fig. 1 is not detailed.In addition, report and/or at least a or multiple epi-position listed are not confirmed as yet by experiment among Fig. 1.Aminoacid sequence about the NY-ESO-1 report is found among the SEQ ID NO:49 in this article.

Fig. 2 shows construct A, and the LAGE-1 that comprises total length NY-ESO-1 and brachymemma is the fusion rotein of LAGE-1a for example.In this embodiment, the N-terminal of the C-terminal of NY-ESO-1 and the LAGE-1 of brachymemma merges together with the Histidine affinity tag, is 288 amino acid whose fusion roteins so that length to be provided.The more details of construct A provide (SEQ ID NO:1, SEQ ID NO:3) in table 1.

Fig. 3 shows construct B, comprise the 3-protein d that do not contain its secretion signal first three/LAGE-1 of one (for example amino acid 20-127), total length NY-ESO-1 and the brachymemma fusion rotein of LAGE-1a for example.In this embodiment, the amino acid/11 27 of 3-protein d merges with the N-terminal of NY-ESO-1, and the N-terminal of the C-terminal of described NY-ESO-1 and the LAGE-1 of brachymemma merges, and is 398 amino acid whose fusion roteins so that length to be provided.The more details of construct B provide (SEQ ID NO:2, SEQ ID NO:4) in part 1.6 in table 1.

Fig. 4 shows construct C, and the LAGE-1 that comprises the NY-ESO-1 of part brachymemma and brachymemma is the fusion rotein of LAGE-1a for example.In this embodiment, the N-terminal of the C-terminal of the NY-ESO-1 of part brachymemma and the LAGE-1 of brachymemma merges, and is 242 amino acid whose fusion roteins so that length to be provided.The more details of construct C provide (SEQ ID NO:5, SEQID NO:7) in table 1.

Fig. 5 shows construct D, comprise the 3-protein d that do not contain its secretion signal first three/NY-ESO-1 of one (for example amino acid 20-approximately or about 127), part brachymemma and the LAGE-1 of the brachymemma fusion rotein of LAGE-1a for example.In this embodiment, the amino acid/11 27 of 3-protein d merges with the N-terminal of the NY-ESO-1 of part brachymemma, and the N-terminal of the C-terminal of the NY-ESO-1 of described part brachymemma and the LAGE-1 of brachymemma merges, and is 352 amino acid whose fusion roteins so that length to be provided.The more details of this embodiment provide (SEQ IDNO:6, SEQ ID NO:8) in table 1.

Fig. 6 shows construct E, and the LAGE-1 that comprises the NY-ESO-1 of brachymemma and brachymemma is the fusion rotein of LAGE-1a for example.In this embodiment, the N-terminal of the C-terminal of the NY-ESO-1 of brachymemma and the LAGE-1 of brachymemma merges, and is 211 amino acid whose fusion roteins so that length to be provided.The more details of construct E provide (SEQ ID NO:9, SEQ ID NO:11) in table 1.

Fig. 7 shows construct F, comprise the 3-protein d that do not contain its secretion signal first three/NY-ESO-1 of one (for example amino acid 20-approximately or about 127), brachymemma and the LAGE-1 of the brachymemma fusion rotein of LAGE-1a for example.In this embodiment, the amino acid/11 27 of 3-protein d merges with the N-terminal of the NY-ESO-1 of brachymemma, and the N-terminal of the C-terminal of the NY-ESO-1 of described brachymemma and the LAGE-1 of brachymemma merges, and is 321 amino acid whose fusion roteins so that length to be provided.The more details of construct F provide (SEQ ID NO:10 in table 1; SEQ ID NO:12).

Fig. 8 shows the alternative embodiment of construct E, i.e. E ', and wherein the N-terminal of the NY-ESO-1 of the C-terminal of the LAGE-1 of brachymemma and brachymemma merges, and is 212 amino acid whose fusion roteins so that length to be provided.The more details of this embodiment construct E ' provide (SEQ IDNO:21 in table 1; SEQ ID NO:23).

Fig. 9 shows construct G, comprise brachymemma NY-ESO-1, brachymemma LAGE-1 for example LAGE-1a and collagen sample zone for example from the fusion rotein in the collagen zone of NY-ESO-1.In this embodiment, the C-terminal in collagen sample zone for example merges with the N-terminal of the LAGE-1 of brachymemma.The N-terminal of the NY-ESO-1 of the C-terminal of the LAGE-1 of brachymemma and brachymemma merges successively, is 289 amino acid whose fusion roteins so that length to be provided.The more details of construct G provide (SEQ ID NO:13 in table 1; SEQ ID NO:15).

Figure 10 shows the synoptic diagram of the exemplary recombinant polypeptide of the NY-ESO-1 that comprises the collagen spline structure territory with part brachymemma.The epi-position that shows among Figure 10-13 only is the representative for the epi-position of this protein report, and is not confirmed by experiment as yet.

Figure 11 shows first three that comprise the 3-protein d that do not contain its secretion signal/one (for example amino acid 20-approximately or about 127) and has the proteic synoptic diagram of exemplary fused of NY-ESO-1 in the collagen spline structure territory of part brachymemma.

Figure 12 shows the synoptic diagram of the exemplary recombinant polypeptide of the NY-ESO-1 that comprises the collagen spline structure territory with part brachymemma.

Figure 13 shows first three that comprise the 3-protein d that do not contain its secretion signal/one (for example amino acid 20-approximately or about 127) and has the proteic synoptic diagram of exemplary fused of NY-ESO-1 in the collagen spline structure territory of brachymemma.

Figure 14 is the synoptic diagram that is presented at many epi-positions of identifying in the LAGE-1a protein of brachymemma.These epi-positions only are that therefore tabulation is not detailed for the representative of the epi-position of this protein report.For fear of query, report and/or the epi-position listed may be confirmed or may do not confirmed by experiment as yet (that is, they may be predicted etc.) by experiment among this figure, unless this paper has explanation in addition.The LAGE-1a aminoacid sequence is listed as SEQ ID NO:58 in sequence table completely.LAGE-1b aminoacid sequence (LAGE-1b does not describe in this drawing) is listed as SEQ ID NO:71 in sequence table completely.

Figure 15 shows NY-ESO-1 and LAGE-1, and the synoptic diagram of many MHC (main histocompatibility complex) I and II class epi-position.These epi-positions only are that therefore tabulation is not detailed for the representative of the epi-position of this protein report; One or more epi-positions of reporting and/or listing are not confirmed as yet by experiment.

Figure 16 shows that NY-ESO-1/LAGE-1 merges the synoptic diagram of design.

Figure 17 has summarized 15 kinds of constructs and production level thereof in a schematic way.The P=3-protein d; C (grey box)=NY-ESO-1 collagen spline structure territory; The collagen spline structure territory of C (white box)=brachymemma; L=does not contain the Lage 1 in collagen spline structure territory; N=does not contain the NY-ESO-1 in collagen spline structure territory; Black arrow mark=polyhistidyl label; (-)=low production; (+)=some productions; (++)=high production; (+++)=best production.Be summarized in table 4 and the sequence table about its nucleotide sequence of the aminoacid sequence of 8 kinds of constructs and coding.

Figure 18 summarizes screening #1; use each the tests in 76 days among the assessment of CB6F1 mouse LVL076, LVL079, LVL78, LVL68, LVL020, LVL26, LVL024, the LVL30, whether give at protection with the subcutaneous attack of transplantation tumor (B16/NYESO1) to measure the intramuscular immunization that adds adjuvant with fusion rotein.

Figure 19 is summarised in the B-16-NY-ESO-1 tumor growth in the control mice of using in the test in 76 days.

Figure 20 shows the survival with the mouse of total length NY-ESO-1, LVL030, LVL068, LVL079 or LVL026 immunization.

Figure 21 summarizes as passes through the NY-ESO-1 specific immune response of ELISA, FACS and western blotting assessment, and as LAGE-1a (the not containing collagen spline structure territory) specific immune response by ELISA and FACS assessment.

Figure 22 summarizes screening #2, and whether the experimental design of test in 105 days is given at B16/NY-ESO-1 and being attacked and the protection of B16/LAGE-1a attack to measure the intramuscular immunization that adds adjuvant with selected fusion rotein.Shown the B16/NY-ESO-1 attack.

Figure 23 has summarized screening #2, and has shown the B16/LAGE-1a attack.

Figure 24 shows that B16/NY-ESO-1 attacks the survival of back with the mouse of LVL078, LVL068, total length NY-ESO-1, LVL024 and LVL076 immunization.Referring to Figure 24.

Figure 25 show B16/LAGE-1a attack the back with LVL076, do not contain the survival of mouse of LAGE-1a, LVL024, total length NY-ESO-1, LVL078 or the LVL068 immunization in collagen sample zone.

Figure 26.Post 1-8 from left to right shows the result of performed ELISAs, to detect possible people's collagen specificity immunne response in the mouse of one of the following immunization: (1) damping fluid (contrast); (2) total length NY-ESO-1; (3) do not contain the LAGE-1a in collagen spline structure territory; (4) LVL068; (5) LVL078; (6) LVL024; (7) LVL076.Positive control (post 8) comprises anti-people's collagen 1 monoclonal antibody (the anti-people's collagen I of mAb).

Embodiment

Fusion rotein.Fusion rotein of the present invention is used for the treatment of cancer, and more specifically is used for the treatment of: melanoma; Mammary cancer; Prostate cancer; Bladder cancer comprises transitional cell carcinoma; Lung cancer comprises nonsmall-cell lung cancer (NSCLC); The H﹠N cancer comprises the esophageal carcinoma; Squamous cell carcinoma; Gastrointestinal cancer; Liver cancer; Cerebral tumor; Leukemia; With various sarcomas.

Based on the expression overview of LAGE-1 and NY-ESO-1, fusion rotein according to the present invention has effective potentiality in the mammary cancer of estimation 37%.Treatment in accordance with the present invention also may be particularly suitable for treatment for the underproof patient of Her2/neu targeted therapy.Fusion rotein of the present invention also is expected among the patient that about 35% patients with prostate cancer, 35% bladder cancer patients, 40% melanoma patient and 35% have NSCLC (nonsmall-cell lung cancer) effectively.In one embodiment, fusion rotein of the present invention can make it possible to treat patient colony widely, can give fusion rotein of the present invention because have the patient of the tumour of expressing NY-ESO-1 and/or LAGE-1 (comprising LAGE-1a and LAGE-1b).

Also can immunogenicity more be arranged than its individual components protein according to fusion rotein of the present invention, this is owing to following reason:

The removal in the one or more collagen spline structure of ■ territory can reduce the potential immunotolerance of the compound that the homology by itself and natural endogenous collagen structure causes, or

The optional interpolation of ■ allos fusion partner can further stimulate cd4 t cell to reply.Therefore, fusion rotein is used to induce for cancer antigen for example NY-ESO-1 or LAGE-1 or both immunogenic responses.

The NY-ESO-1 of Shi Yonging can be total length, the NY-ESO-1 part brachymemma or brachymemma or its any fragment in the present invention, and described fragment comprises one or more epi-positions that can cause for the immunne response of NY-ESO-1.Total length NY-ESO-1 protein in the background of this specification sheets means approximately or about 1-180 amino acid, and with naturally occurring protein (SEQ ID NO:49) at least 95,96,97,98,99% or 100% protein that is equal to.As used herein, term " LAGE-1 " refers to one or more LAGE-1 family members, and for example LAGE-1a and LAGE 1b are as mentioned below." total length LAGE-1a " protein mean with SEQ ID NO:58 be 95,96,97,98,99% or 100% protein that is equal to.Similarly, " total length LAGE-1b " protein means and naturally occurring protein (SEQ ID NO:71) 95,96,97,98,99% or 100% protein that is equal to.

In one embodiment, identity is on the total length of sequence.Therefore, the present invention also extends to the described fusion rotein with conservative substitution.Conservative substitution is well-known, and generally sets up according to the default rating matrix in the sequence alignment computer program.These programs comprise PAM250 (people such as Dayhoft M.O., (1978), " A model of evolutionary changes inproteins ", " Atlas of Protein sequence and structure " 5 (3) M.O.Dayhoft (ed.), 345-352), National Biomedical Research Foundation, Washington, with Blosum 62 (Steven Henikoft and Jorja G.Henikoft (1992), with " Amino acidsubstitution matricies from protein blocks "), Proc.Natl.Acad.Sci.USA 89 (Biochemistry): 10915-10919.

Generally speaking, the displacement in following group is a conservative substitution, but displacement is regarded as nonconservative between group.Group is:

I) aspartic acid/l-asparagine/L-glutamic acid/glutamine,

Ii) serine/threonine,

Iii) Methionin/arginine,

Iv) phenylalanine/tyrosine/tryptophane,

V) leucine/Isoleucine/Xie Ansuan/methionine(Met),

Vi) glycine/L-Ala.

" the part brachymemma " in this specification sheets background means NY-ESO-1 or LAGE-1 protein (suitably time), wherein most of collagen sample zone is removed, but still comprises the epi-position A31 that finds in this zone or be made up of the epi-position A31 that finds in this zone.

In one embodiment, the NY-ESO-1 of part brachymemma and/or LAGE-1 comprise following amino acid scope or be made up of following amino acid scope: amino acid 44,45,46,47,48,49,50,51 or 52 is to amino acid/11 75,176,177,178,179 or 180, or these amino acid whose any combinations, for example amino acid 48-amino acid/11 80 or amino acid 46-amino acid/11 78.In one embodiment, the NY-ESO-1 of part brachymemma or LAGE-1 comprise approximately or accurately amino acid 48-180 (or under the situation of LAGE-1b, amino acid 48-210 approximately or accurately), or form by amino acid 48-180 approximately or accurately (or under the situation of LAGE-1b, approximately or accurately amino acid 48-210).In one embodiment, the term " about " in this background can be used in reference to the amino acid that is up to sequence amino acid overall number+/-10% and choose interpolation or disappearance from sequence wantonly.In one embodiment, the NY-ESO-1 of part brachymemma comprises the amino acid 48-180 of NY-ESO-1, or is made up of the amino acid 48-180 of NY-ESO-1.

In one embodiment, the LAGE-1b of part brachymemma comprises following amino acid scope or is made up of following amino acid scope: amino acid 44,45,46,47,48,49,50,51 or 52 is to amino acid 205,206,207,208,209 or 210, or these amino acid whose any combinations, for example amino acid 48-amino acid 210 or amino acid 46-amino acid 208.In one embodiment, the LAGE-1b of part brachymemma comprises approximately or amino acid 48-210 accurately, or is made up of amino acid 48-210 approximately or accurately.In one embodiment, the term " about " in this background can be used in reference to the amino acid that is up to sequence amino acid overall number+/-10% and choose interpolation or disappearance from sequence wantonly.In one embodiment, the LAGE-1b of part brachymemma comprises the amino acid 48-210 of LAGE-1b, or is made up of the amino acid 48-210 of LAGE-1b.

" brachymemma " in this specification sheets background means NY-ESO-1 or LAGE-1 protein (suitably time), and wherein collagen sample zone is removed (comprising the removal of A31 epi-position).In one embodiment, the NY-ESO-1 of brachymemma and/or LAGE 1 comprise approximately or accurately amino acid 71-180 (or under the situation of LAGE-1b, amino acid 71-210 approximately or accurately), or form by amino acid 71-180 approximately or accurately (or under the situation of LAGE-1b, approximately or accurately amino acid 71-210).

In one embodiment, the NY-ESO-1 of brachymemma or LAGE-1 comprise following amino acid scope or be made up of following amino acid scope: amino acid 67,68,69,70,71,72,73,74 or 75 is to amino acid/11 75,176,177,178,179 or 180, or these amino acid whose any combinations, for example amino acid 71-amino acid/11 80 or amino acid 69-amino acid/11 78.In one embodiment, the NY-ESO-1 of brachymemma or LAGE-1 comprise approximately or accurately amino acid 71-180 (or under the situation of LAGE-1b, amino acid 71-210 approximately or accurately), or form by amino acid 71-180 approximately or accurately (or under the situation of LAGE-1b, approximately or accurately amino acid 71-210).

In one embodiment, the term " about " in this background can be used in reference to the amino acid that is up to sequence amino acid overall number+/-10% and choose interpolation or disappearance from sequence wantonly.In one embodiment, the NY-ESO-1 of brachymemma or LAGE-1 comprise the amino acid 71-180 of NY-ESO-1 or LAGE-1, or are made up of the amino acid 71-180 of NY-ESO-1 or LAGE-1.

In one embodiment, the LAGE-1b of brachymemma comprises following amino acid scope or is made up of following amino acid scope: amino acid 67,68,69,70,71,72,73,74 or 75 is to amino acid 205,206,207,208,209 or 210, or these amino acid whose any combinations, for example amino acid 71-amino acid 210 or amino acid 69-amino acid 208.In one embodiment, the LAGE-1b of brachymemma comprises approximately or amino acid 71-210 accurately, or is made up of amino acid 71-210 approximately or accurately.In one embodiment, the term " about " in this background can be used in reference to the amino acid that is up to sequence amino acid overall number+/-10% and choose interpolation or disappearance from sequence wantonly.In one embodiment, the LAGE-1b of brachymemma comprises the amino acid 71-210 of LAGE-1b, or is made up of the amino acid 71-210 of LAGE-1b.

" other fragments " means such fragment, and in the time of in mixing fusion rotein of the present invention, it causes having the final protein of the required character and the advantage of fusion rotein of the present invention.

NY-ESO-1。Provide the antigenic modified antigen that comprises derived from Tumor rejection antigen NY-ESO-1 according to preamble, wherein the collagen zone is part brachymemma or brachymemma fully.In some embodiments, remove above the collagen zone.In some embodiments, modified antigen is a genetic modification.In some embodiments, modified antigen is a recombinant chou.In some embodiments, provide and comprised the antigenic polypeptide of describing in the sentence as described above.In some embodiments, exemplary polypeptide comprises heterologous protein, for example from 3-protein d or its fragment of Haemophilus influenzae (Haemophilus influenzae) Type B.The construct of the nucleotide sequence that comprises the aforementioned polypeptide of encoding is provided in some embodiments.

In some embodiments, provide to comprise NY-ESO-1 or its segmental immunogenic polypeptide, wherein NY-ESO-1 does not comprise collagen sample zone.In other embodiments, NY-ESO-1 is the part brachymemma or brachymemma or comprises its any fragment that described fragment comprises one or more epi-positions.In some embodiments, this peptide species has conservative substitution.In some embodiments, this peptide species and construct are used for prevention or improve cancer return basically as prophylactic agent.

Therefore, in some embodiments, one or more amino acid are removed from the collagen zone.More specifically, in some embodiments, 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72 or 73 amino acid are removed from the part that comprises the collagen zone, i.e. the amino acid/11 of SEQ ID NO:49-73 roughly.Amino acid can be removed from the close position the collagen zone or from non-close position.In other words, in some embodiments, amino acid in that part of SEQ ID NO:49 following any position or its any combination remove: 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72 or 73.It will be appreciated by those skilled in the art that in some embodiments the part of aminoacid sequence is preserved like this, thereby make defined epitope wherein be retained in the resulting polypeptide.

In some embodiments, utilize the fragment in NY-ESO-1 middle section or C-terminal zone.Therefore, in some embodiments, polypeptide can comprise one or more fragments of aminoacid sequence shown in the SEQ ID NO:49, promptly comprises the fragment of one or more or its any combination among amino acid position 74-75,76-80,81-85,86-90,91-95,96-100,101-105,106-110,111-115,116-120,121-125,126-130,131-135,136-140,141-145,146-150,151-155,156-160,161-165,166-170,171-175, the 176-180.It will be appreciated by those skilled in the art that in some embodiments aminoacid sequence is preserved like this, thereby make defined epitope be retained in the resulting polypeptide.

LAGE-1。In some embodiments, provide the antigenic modified antigen that comprises derived from Tumor rejection antigen LAGE-1, wherein the collagen zone is part brachymemma or brachymemma fully.In some embodiments, remove above the collagen zone.In some embodiments, modified antigen is a genetic modification.In some embodiments, modified antigen is a recombinant chou.In some embodiments, provide and comprised the antigenic polypeptide of describing in the sentence as described above.In some embodiments, antigen is derived from Tumor rejection antigen LAGE-1a.In some embodiments, antigen is derived from Tumor rejection antigen LAGE-1b.In other embodiments, exemplary fused albumen comprises heterologous protein, for example from 3-protein d or its fragment of Haemophilus influenzae Type B.The construct of the nucleotide sequence that comprises the aforementioned polypeptide of encoding is provided in some embodiments.

In some embodiments, provide to comprise LAGE-1 or its segmental immunogenic polypeptide, wherein LAGE-1 does not comprise collagen sample zone.In other embodiments, LAGE-1 is the part brachymemma or brachymemma or comprises its any fragment that described fragment comprises one or more epi-positions.In some embodiments, polypeptide comprises the hybrid in the collagen sample zone of LAGE-1 polypeptide and NY-ESO-1.In some embodiments, polypeptide comprises the part or all of NY-ESO-1 collagen zone that is connected with the LAGE-1 part brachymemma or brachymemma.In some embodiments, this peptide species has conservative substitution.In some embodiments, this peptide species and construct are used for prevention or improve cancer return basically as prophylactic agent.

Therefore, in some embodiments, one or more amino acid are from the collagen zone or even remove from N-terminal amino acid.More specifically, in some embodiments, 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72 or 73 amino acid from the collagen zone or even N-terminal amino acid remove the i.e. amino acid/11-73 of SEQ ID NO:58 (LAGE-1a) or SEQ ID NO:71 (LAGE-1b) roughly.Amino acid can be removed from the close position this zone or from non-close position.In other words, in some embodiments, one or more amino acid in SEQ ID NO:58 (LAGE-1a) or SEQ ID NO:71 (LAGE-1b) following any position or its any combination remove: 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72 or 73.It will be appreciated by those skilled in the art that in some embodiments the part of aminoacid sequence is preserved like this, thereby make defined epitope wherein be retained in the resulting polypeptide.

In some embodiments, utilize the fragment in LAGE-1 middle section or C-terminal zone.Therefore, in some embodiments, polypeptide can comprise one or more fragments of aminoacid sequence shown in SEQ ID NO:58 (LAGE-1a) or the SEQ ID NO:71 (LAGE-1b), promptly comprises amino acid position 74,75,76,77,78,79,80,81-85,86-90,91-95,96-100,101-105,106-110,111-115,116-120,121-125,126-130,131-135,136-140,141-145,146-150,151-155,156-160,161-165,166-170,171-175, the fragment of one or more or its any combination among the 176-180.It will be appreciated by those skilled in the art that in some embodiments aminoacid sequence is preserved like this, thereby make defined epitope be retained in the resulting polypeptide.

In one aspect, the invention provides the fusion rotein that comprises total length NY-ESO-1.

In one aspect, the invention provides the fusion rotein of the NY-ESO-1 that comprises the part brachymemma.

In one aspect, the invention provides the fusion rotein of the NY-ESO-1 that comprises brachymemma.

In one aspect, the invention provides the fusion rotein that comprises total length LAGE-1.

In one aspect, the invention provides the fusion rotein of the LAGE-1 that comprises the part brachymemma.

In one aspect, the invention provides the fusion rotein of the LAGE-1 that comprises brachymemma.

In one aspect, the LAGE-1 that uses in the present invention is LAGE-1a.

In one aspect, the LAGE-1 that uses in the present invention is LAGE-1b.

In one aspect of the invention, the C-terminal of the N-terminal of NY-ESO-1 and LAGE-1 merges.

In one aspect of the invention, the N-terminal of the C-terminal of NY-ESO-1 and LAGE-1 merges.

By mixing fragment from further heterologous antigen, 3-protein d for example, the surface protein of gram negative bacterium Haemophilus influenzae B, the immunogenicity of fusion rotein of the present invention can further increase and/or proteinic nature of production can further improve.Can derive from WO 91/18926 about more information derived from the immunology fusion partner of 3-protein d.

The protein that is used for being included in fusion partner of the present invention can carry out chemically conjugated, or can be used as recombination fusion protein and express.In one embodiment, fusion rotein is expressed as recombination fusion protein.

Further the allos fusion partner can help to provide T to assist epi-position (immunology fusion partner), maybe can help with higher output yield marking protein (expression enhanser).In one embodiment, further the allos fusion partner can be the immunology fusion partner be again to express to strengthen mating partner.

In one embodiment, the 3-protein d or derivatives thereof comprises approximately or is accurately proteinic preceding 1/3, for example, and pact or the amino acid/11 of 3-protein d-109 accurately.In this embodiment, the amino acid 2-Lys of natural protein D sequence and/or 3-Thr can be by amino acid 2-Asp and/or 3-Pro displacements.In further embodiment, the 3-protein d or derivatives thereof comprises approximately or the amino acid 20-127 of 3-protein d accurately, or is made up of the amino acid 20-127 of 3-protein d approximately or accurately.In one embodiment, the 3-protein d that is used for using does not in the present invention comprise proteinic secretion sequence.Usually, in fusion rotein of the present invention, the 3-protein d derivative is not a lipidization.

In one embodiment, 3-protein d for example further comprises amino acid Met, Asp and the Pro (be that construct can comprise " MDP-20-127 3-protein d ", or be made up of " MDP-20-127 3-protein d ") that merges with the segmental N-terminal of 3-protein d.Think that these 3 kinds of other amino acid can help stabilizing protein and/or increase its protein expression level.

In one aspect, the invention provides fusion rotein, wherein the N-terminal fragment of 3-protein d (as mentioned above) (be first three/) merges with the N-terminal of fusion rotein of the present invention or its immunogenic fragments.More specifically, the fusions of the N-terminal of 3-protein d and fusion rotein of the present invention can be realized like this, thereby make the latter substitute the C-terminal fragment of cut 3-protein d.Therefore, the N-terminal of 3-protein d becomes the N-terminal of fusion rotein.

Other allos fusion partners or its fragment can be included in the fusion rotein of the present invention, and replace 3-protein d or add 3-protein d, for example:

■ is from the Nonstructural Protein of influenza virus, NS1 (hemagglutinin).Usually, can use 81 amino acid of N-terminal, although can also use different fragments, prerequisite is that they comprise that T assists epi-position;

■ is derived from the LytA of streptococcus pneumoniae (Streptococcus pneumoniae), it is synthetic by LytA gene (Gene, 43 (1986) 265-272 pages or leaves) Bian Ma N-acetyl-L-ala amide enzyme LytA, the repeating part of the LytA molecule of for example finding in C-terminal for example arises from for example residue 188-305 of residue 178.In one embodiment, the allos fusion partner is CLytA.In further embodiment, the allos fusion partner is CPC, comprises the fusion rotein of CLytA-P2-CLytA, described in WO03/104272.The purifying that comprises the segmental hybrid protein matter of C-LytA at its N-terminal place obtains in (1992) the 795-798 pages or leaves describing at Biotechnology:10.

Fusion rotein of the present invention may further include affinity tag, for example comprises 1-10 for example Histidine tail of 6 or 10 histidine residues (being also referred to as the his-label).These residues can be for example endways on the part, for example proteinic N-terminal and/or C-terminal part.Can mix affinity tag, with the further protein purification that improves.

Concrete fusion roteins more of the present invention can for example make up described in accompanying drawing.Each embodiment shown in the accompanying drawing is represented independent aspects of the present invention.According to more many cases of the construct of fusion rotein of the present invention in table 1-4 and provide in the sequence table.

Nucleic acid.The present invention also extends to nucleic acid and Polynucleotide (polynucleic acid), for example DNA of coding fusion rotein of the present invention.Method of the present invention can be undertaken by conventional recombinant technology, people such as Maniatis for example, Molecular Cloning-A Laboratory Manual; Cold Spring Harbor describes among the 1982-1989.Especially, method can comprise the steps:

I) preparation can be expressed the reproducible or integrating expression vector of the DNA polymkeric substance that comprises nucleotide sequence, described nucleotide sequence coded fusion rotein or its immunogenic derivatives in host cell;

Ii) use described carrier transformed host cell;

Iii), cultivate described transformed host cells allowing to express described DNA polymkeric substance to produce under the described proteinic condition; With

Iv) reclaim described protein.

Term ' conversion ' is used in reference to foreign DNA in this article and introduces in the host cell.This can wherein use for example as Genetic Engineering for example by reaching with suitable plasmid or virus vector conversion, transfection or infection; Editor S.M.Kingsman and A.J.Kingsman; BlackwellScientific Publications; Oxford, England, the routine techniques of describing in 1988.Term ' conversion ' or ' transformant ' will be applied to comprise and express the resulting host cell of purpose alien gene hereinafter.The expression vector that comprises the nucleotide sequence of the fusion rotein of the present invention of encoding is new, and constitutes part of the present invention.

Reproducible expression vector can be prepared according to the present invention, this is by cutting the carrier compatible with host cell so that the section of the linear DNA with complete copy to be provided, and described linear segments and one or more dna moleculars are made up to be realized, described dna molecular is together with the described linear fragment required product of encoding, the DNA polymkeric substance of the protein or derivatives thereof of the present invention of for example encoding.Therefore, when needing, the DNA polymkeric substance can be pre-formed or form in the building process of carrier.

The selection of carrier will part by the host cell decision, described host cell can be protokaryon or eucaryon, but generally be intestinal bacteria or Chinese hamster ovary celI.Suitable carriers comprises plasmid for example TMCP14 or pET21 or pET26, pcDNA3, phage, clay and recombinant virus.In one embodiment, wherein be expressed in baculovirus, yeast or the CHO host cell, can use one of following carrier: pEE14, pPICZA, pPICZB, pPICZC, pDMT-DEST48 and pAcSG2.Reproducible preparation of expression vectors can be with carrying out about DNA restriction, polymerization and the suitable enzymes that is connected routinely, the program of describing by the philtrums of for example above quoting such as Maniatis.

By under conversion condition with reproducible expression vector transformed host cell of the present invention, prepare recombinant host cell according to the present invention.Suitable conversion condition is conventional, and the people such as Maniatis that for example above quote, or " DNA Cloning " II volume, and D.M.Glover edit, IRLPress Ltd, description in 1985.The selection of conversion condition is determined by host cell.Therefore, host bacterium for example intestinal bacteria (E.coli) can be used CaCl2 solution (people such as Cohen, Proc.Nat.Acad.Sci., 1973,69,2110), or comprises RbCl, MnCl ₂, potassium acetate and glycerine the solution of mixture handle, and use the 3-[N-morpholino then]-propanesulfonic acid, RbCl and glycerine handles.Mammalian cell in the cultivation can be by transforming the co-precipitation of carrier DNA calcium to cell.The present invention also extends to reproducible expression vector transformed host cells of the present invention.

DNA can carry out the codon optimization by standard technique, with the relevant host's of further promotion expression.

Under the condition that allows the DNA polymkeric substance to express, cultivate transformed host cells and carry out routinely, described in people such as the Maniatis that for example above quotes and " DNA Cloning ".Therefore, preferably provide nutrient substance, and cultivate in the temperature that is lower than 50 ℃ to cell.Protein of the present invention can for example be expressed in the yeast prokaryotic organism or eukaryote, but expresses in intestinal bacteria usually.Can use colibacillary specific bacterial strain, for example:

■ AR58: derived from the hiding λ lysogen of N99, it is gal E::Tn 10, Δ-8 (chlD-pgl), Δ-H1 (cro-chlA), N ⁺And cI857 (reference: Proc.Natl.Acad.Sci.USA the 82nd volume, 88-92 page or leaf, January 1985 Biochemistry)

■ BLR (DE3) Novagen, WI, (catalog number (Cat.No.): 69053-4): BLR is the recA-derivative of BL21 to USA.Usually, mix the selective marker of kalamycin resistance for example or amicillin resistance, to promote successfully to mix the evaluation of the recombination/construct in the expression system.

Product reclaims according to host cell and according to the location of expression product (in the cell or be secreted in the substratum or in the cell pericentral siphon) by ordinary method.Therefore, when host cell is a bacterium for example during intestinal bacteria, it is physics, chemistry or enzymatic lysis for example, and from resulting lysate the isolated protein product.When host cell is Mammals, generally can be from nutritional medium or from cell-free extract separated product.Conventional protein stripping technique comprises selective precipitation, adsorption chromatography and comprises the affinity chromatography of monoclonal antibody affinity column.

Soluble or lyophilized form provides protein of the present invention with liquid form.The present invention also provides the pharmaceutical composition that comprises fusion rotein of the present invention and pharmaceutically acceptable vehicle, for example vaccine.

When using, therapeutic composition of the present invention can be used with pharmaceutically acceptable preparation.This kind preparation can comprise salt, buffer reagent, sanitas, compatible carrier, for example adjuvant and the cytokine and the randomly other treatment agent of complementarity immunostimulant of pharmaceutically acceptable concentration routinely.

Amount will depend on concrete situation to be treated certainly, the severity of situation, the individual patient parameter comprises age, physical appearance, size and weight, the treatment time length, the character of simultaneous treatment (if having any treatment), particular route of administration and in healthy practitioner's knowledge and the similar factor in the expertise.These factors are that those of ordinary skills are well-known, and can solve by being no more than routine test.The maximal dose of general preferred use individual components or its combination, the i.e. the highest safe dose of judging according to rational medicine.Yet, it will be appreciated by the skilled addressee that owing to medical reasons, psychology reason or owing to any other reason basically, the patient can adhere to than low dosage or tolerance dose.Be generally expected that everyone dosage will comprise 1-1000 μ g protein, and preferred 30-300 μ g.

In one aspect, the pharmaceutical composition that is used to use fusion rotein of the present invention will be a vaccine.Vaccine can be chosen wantonly and comprise one or more other tumor associated antigens, polypeptide and/or peptide.For example, the member who belongs to MAGE, LAGE and GAGE family.

The combination of NY-ESO-1/LAGE-1 and MAGE.In one embodiment of the invention, provide and comprised following composition: (a) comprised the antigen component of NY-ESO-1 as described herein or LAGE-1 antigen or fusion rotein and (b) comprise the antigen component of MAGE antigen or fusion rotein.In one embodiment, composition can further comprise adjuvant as described herein.

The MAGE antigen that is used for using in combination can comprise total length MAGE antigen.Alternately, MAGE antigen can comprise the immunogenicity part of MAGE, wherein 1,2,3,4,5,6,7,8,9,10 or more amino acids can replace from sequential amino acid deletion or aminoacid sequence.In one embodiment of the invention, 2 amino acid can be from MAGE sequence of N terminal deletion.In one embodiment of the invention, wherein antigen is MAGE-A3 or its immunogenicity part, and the sequence of MAGE-A3 can be from the amino acid 3-314 of MAGE-A3.

In one embodiment of the invention, provide the composition that comprises NY-ESO-1/LAGE-1 antigen as described herein and/or fusion rotein and comprise the antigenic fusion rotein of MAGE-A3.In alternative embodiment, comprise preceding 109 amino acid whose fusion partner protein that the antigenic fusion rotein of MAGE-A3 comprises MAGE-A3 antigen and comprises about approximately or about 3-protein d, or by MAGE-A3 antigen with comprise approximately preceding 109 amino acid whose fusion partner protein about or about 3-protein d and form, wherein one or two or more amino acid from 3-protein d are optional metathetical, and wherein outside preceding 109 amino acid of isolating protein D, choose the signal sequence that has 3-protein d wantonly.

Fusion rotein of the present invention can be chosen wantonly in addition and comprise one or more amino acid, as between antigen and fusion partner or the fusion partner protein sequence or " joint " between antigen and the His tail (if present).Amino acid can be irrelevant with the sequence of antigen and/or fusion partner.

As described herein, fusion rotein of the present invention can be additionally contained in the amino acid Met-Asp-Pro of fusion rotein sequence of N end.Met amino acid can be from urporotein D sequence or can be from irrelevant sequence.

In one embodiment, be used for being shown among the SEQ ID NO:98 in the fusion rotein sequence that comprises MAGE-A3 and 3-protein d that combination of the present invention is used.SEQ ID NO:98 comprises following key element from N-terminal:

Amino acid/11-18 comprises 1-Met and to the signal sequence of the 3-protein d of the displacement 2-Asp of the natural amino acid 2-Lys of 3-protein d and 3-Thr and 3-Pro

Amino acid/11 9-127 comprises the amino acid 20-127 of 3-protein d

Amino acid/11 28-129 at the irrelevant amino acid Met-Asp at amino acid/11 28-129 place to produce cloning site

The fragment of amino acid/11 30-441 MAGE3 (the amino acid 3-314 of MAGE3)

The amino acid 442-443 amino acid Gly-Gly that has nothing to do

Amino acid 444-451 7his tail

The present invention also extends to the described vaccine/method for compositions of preparation, and maybe can be by the fusion rotein and the vaccine/composition of described method acquisition by described method acquisition.

Vaccine production is generally at Vaccine Design (" The subunit and adjuvantapproach " (editor Powell M.F.﹠amp; Newman M.J). (1995) Plenum Press NewYork) the middle description.Describe by Fullerton United States Patent (USP) 4,235,877 at the intravital encapsulation of lipid.

Fusion rotein of the present invention can be in vaccine preparation of the present invention adjuvantization.Suitable adjuvant comprises aluminium salt for example aluminum hydroxide gel (alum (alum)) or aluminum phosphate, but also can be calcium, iron or zinc salt, maybe can be the polysaccharide of acidylate tyrosine or acidylate sugar, positively charged ion or anionic derivative or the insoluble suspension of polyphosphonitrile.Other known adjuvants comprise the oligonucleotide that contains CpG.Oligonucleotide is characterised in that the CpG dinucleotides is unmethylated.This kind oligonucleotide is well-known, and is for example describing among the WO 96/02555.

In preparation of the present invention, may wish that adjunvant composition preferentially induces the immunne response of TH1 type.In one embodiment, provide comprise that for example the preferred 3-of monophosphoryl lipid A takes off-O-acidylate monophosphoryl lipid A (3D-MPL) is together with the adjuvant system of the combination of aluminium salt.The CpG oligonucleotide also can induce TH1 to reply, and also can be comprised.

In one embodiment, composition is provided, described composition comprises fusion rotein as described herein, with the combination that comprises monophosphoryl lipid A and saponin derivative, particularly as the adjunvant composition of the combination of disclosed QS21 and 3D-MPL among the WO94/00153, or as the low reaction originality composition of disclosed QS21 among the WO96/33739 by the cholesterol quencher.Operable a kind of preparation comprises, QS21,3D-MPL and tocopherol in the oil-in-water emulsion of describing in WO 95/17210 for example.The another kind of adjuvant formulation that is used for using in the present invention for example can comprise oil-in-water emulsion or as QS21,3D-MPL and CpG or its Equivalent of Liposomal formulation.Therefore, in one embodiment of the invention, provide the vaccine that comprises fusion rotein of the present invention and adjuvant, for example as mentioned above.The present invention also extends to vaccine and the method for compositions that preparation comprises fusion rotein as described herein.

The present invention expects that also nucleic acid as described herein, polypeptide or peptide are used for vaccinated sending.Sending of polypeptide and peptide can be finished according to standard inoculation vaccine rules well-known in the art.In another embodiment, sending of nucleic acid can be finished by method in the body of earlier external back, promptly by from the experimenter, taking out cell, make cytogene engineered comprising cancer associated antigens, and engineered cell is introduced in the experimenter again.Generally speaking, this function that can relate to gene copies in external introducing experimenter's the cell, and genetic engineering modified cell is sent back in the experimenter.The function copy of gene is under the control operated of regulatory element, and this allows gene to express in genetic engineering modified cell.Numerous transfections and transduction technology and suitable expression vector are that those of ordinary skills are well-known, and some of them are described in PCT application WO95/00654.Also expect according to the present invention use carrier for example the nucleic acid in vivo of the liposome of virus and target send.

Abbreviation

CO collagen sample zone

W/Ocoll does not contain collagen sample zone (collagen spline structure territory)

The PD1/3 3-protein d first three/one

Its exemplary of nucleotide sequence of fusion rotein and coding provides among the 1-3 at table.

Table 1.Provide fusion rotein and coding its exemplary of nucleotide sequence.Every kind of nucleotide sequence is identified by the nucleotides sequence column identifier (SEQ IDNO :) of uniqueness, and is set forth in sequence table by subject description.Every kind of fusion rotein is identified by the aminoacid sequence identifier (SEQ ID NO :) of uniqueness, and is set forth in sequence table by subject description.

Table 2.Provide fusion rotein and coding its other exemplary of nucleotide sequence.Every kind of nucleotide sequence is identified by the nucleotides sequence column identifier (SEQID NO :) of uniqueness, and is set forth in sequence table by subject description.Every kind of fusion rotein is identified by the aminoacid sequence identifier (SEQ ID NO :) of uniqueness, and is set forth in sequence table by subject description.

Table 3.Provide fusion rotein and coding its other exemplary of nucleotide sequence.Every kind of nucleotide sequence is identified by the nucleotides sequence column identifier (SEQID NO :) of uniqueness, and is set forth in sequence table by subject description.Every kind of fusion rotein is identified by the aminoacid sequence identifier (SEQ ID NO :) of uniqueness, and is set forth in sequence table by subject description.

Table 4.Fusion rotein of discussing among the embodiment and the nucleotide sequence of encoding it are provided.Every kind of nucleotide sequence is identified by the nucleotides sequence column identifier (SEQ IDNO :) of uniqueness, and is set forth in sequence table by subject description.Every kind of fusion rotein is identified by the aminoacid sequence identifier (SEQ ID NO :) of uniqueness, and is set forth in sequence table by subject description.

As will be conspicuous by sequence table, many constructs of table 4 have and the similar design of one or more embodiments shown in the aforementioned table.For example, LVL068 share with table 1 in as the identical design of embodiment shown in the SEQ ID NO:45.LVL076 share with table 1 in as the identical design of embodiment shown in the SEQ ID NO:25.LVL078 share with table 1 in as the identical design of embodiment shown in the SEQ ID NO:33.LVL079 share with table 1 in as the identical design of embodiment shown in the SEQ ID NO:37.

In addition, several fusion protein construct shown in the table 4, be LVL155, LVL106, LVL156, LVL157, LVL151, by other fusion rotein sequences shown in the table 4, the routine that promptly is respectively LVL068, LVL030, LVL076, LVL078, LVL024 is modified and is produced.This kind modification is included in the removal of the amino-acid residue between the beginning of 3-protein d and mosaic (promptly derived from part arbitrary among NY-ESO-1 and the LAGE-1), and the amino acid whose removal between his-label and mosaic begin.Therefore, some fusion rotein of table 4 is closely corresponding to other fusion roteins in the table 4.Correspondence between these fusion roteins is set forth in table 5, and describes in more detail in embodiment 4.

Table 5.Correspondence between the LVL155 of LVL068, LVL030, LVL076, LVL078, LVL024 and modification, LVL106, LVL156, LVL157, the LVL151.

Embodiment

Design of embodiment 1.NY-LA1 chimeric protein and production

As summarizing several NY-ESO-1/LAGE-1 fusion roteins that design contains and do not contain collagen spline structure territory and contain and do not contain the end of 3-protein d among Figure 17.The construct of the design just expression in intestinal bacteria (Escherichia coli) carries out the codon optimization.Synthetic gene is assembled by oligonucleotide and/or PCR product.Use respectively that the KpnI in 5 of optimization gene ' end and 3 ' end and SacI restriction site add NdeI and XhoI site, fragment cloning is arrived in the pGA4 main chain (AmpR).

Plasmid DNA purification from the bacterium that transforms, and by UV spectrometry mensuration concentration.Final construct is verified by order-checking.Use NdeI and XhoI restriction site, will be to pET19 (AmpR) multiple clone site in, with acquisition NY/LAGE chimeric expression plasmid about the direct subclone of optimized encoding sequence of different N Y/LAGE chimeric construct body.For being cloned in the pET26, design PCR primer is to add N-terminal Histidine tail.This amplification causes the interpolation with the in-phase 6 Histidine tails in the coding region of different constructs.This amplified fragments carries out enzymatic digestion with the NdeI/XhoI restriction enzyme, and different NY/LAGE chimeric construct bodies is cloned in pET26 (KanR) multiple clone site in turn, to obtain expression plasmid.Final construct is verified by order-checking.

Shake bottle production.The growth of bacterial host strains and inducing

Cultivate

Bacterium shakes in the bottle at 800ml Luria-Bertani (LB) liquid nutrient medium (BD)+1% (w/v) glucose (Laboratoire MAT at 2.5L, catalog number (Cat.No.): GR-0101)+upward growth of microbiotic (Pyocianil 100 μ g/ml are used for pET19, and kantlex 40 μ g/ml are used for pET26).For cell BLR (DE3), culture in 37 ℃ of incubations until O.D. _600nmAbout 0.8.

Induce

At OD. _600nmAbout 0.8 o'clock, culture BLR (DE3) 1mM isopropyl ss-D-1 sulfo-galactopyranose glycosides (IPTG; EMD Chemicals Inc., catalog number (Cat.No.): 5815) induce, and in 16 ℃ of incubation 16-18 hours.Obtained 5-15mg specified protein/800ml with construct LVL106,151,155 and 157.Protein production about every kind of construct is summarized among Figure 17.

The summary of embodiment 2. preliminary purifications and stability

Proteinic extraction and purifying

Cell is gathered in the crops by centrifugal, breaks by physics and chemical process then, and keeps resulting crude extract, to separate desired polypeptides.

Purifying

Expressed recombinant protein dissolves with guanidine hydrochloride solution, and is loaded into immobilized metal affinity chromatography (Immobilized Metal Affinity Chromatography) (IMAC) on the resin.By before the cumulative imidazole concentration wash-out, use the protein on 8M and the 4M urea soln washing column subsequently.Protein carries out desalination to be used for further use subsequently at last 4M urea buffer solution among the pH7.0.Purifying is assessed by SDS PAGE and western blotting, to verify proteinic purity and identity.

The stability test of the fusion rotein of purifying

Stability is measured in 37 ℃ and is carried out, and protein is assessed by SDS-PAGE.Preliminarily stabilised mensuration does not disclose big problem.

Preliminarily solubilised is measured

The generalized assessment in proteinic solubleness such as the following table.

Chart 1.Fusion rotein solubleness.Keyword: P precipitation; S does not have precipitation; NT does not test.

Embodiment 3: with fusion rotein IM immunization

As described below, fusion rotein carries out clinical preceding assessment in mouse model, relate to a series of intramuscular immunization screening experiments.Selected mouse model is CB6F1, by the first-generation of C57BL6 mouse and Balb/c mouse hybridization generation.This kind mouse is from Charles River Laboratories, Inc., and 251 Ballardvale Street, Wilmington, MA 01887-1000 is obtained commercially.Selected tumor cell line is B16 (a mouse black-in tumor cell system), is used to study the portable muroid melanoma that is obtained commercially of cancer therapy.

Screening #1

Experimental design.In test in 76 days; whether the CB6F1 mouse is used for assessing each of LVL076, LVL079, LVL078, LVL068, LVL020, LVL026, LVL024, LVL030, give at the protection with the subcutaneous attack of the tumour cell of transplanting (B16/NYESO1) to measure the intramuscular immunization that adds adjuvant with fusion rotein.Particularly, mouse carries out the intramuscular immunization with the 50 μ L injections that comprise 15 μ g protein and adjuvant.Selected adjuvant is AS15.AS15 is the liposome adjuvant formulation that comprises QS21,3D-MPL and CpG.

Test with the fusion rotein shown in 1A and the 1B hereinafter.The group that mouse is divided into 15 mouse/groups.Mouse is following in the time of the 0th day and carried out immunization once more in the time of the 14th day:

Test 1A

■LVL079

■LVL026

■LVL068

■LVL030

Test 1B

■LVL076

■LVL020

■LVL078

■LVL024

Contrast

■ antigen damping fluid/AS15 damping fluid

■ total length NY-ESO-1

■ does not contain the LAGE-1a in collagen spline structure territory (CLD)

■MAGE A3

6 mouse/groups were attacked with the B16/NY-ESO-1 tumour of subcutaneous transplantation in the time of the 28th day.In the time of the 0th, 14,28 and 76 day, assess for the NY-ESO-1 total length, do not contain the LAGE-1a in collagen spline structure territory and the antibody response of people's collagen by ELISA (IgG1 and IgG2a).The splenocyte that used results in the time of the 28th day is by cell-mediated the replying of FACS assessment (stimulate-3 storehouses again with NY-ESO-1 and LAGE-1a peptide storehouse, every storehouse 3 (3 pools of 3)).The experimental design of screening #1 is summarized among Figure 18.

The result.In 4 contrasts, compare with damping fluid, only total length NY-ESO-1 gives some protections.Referring to Figure 19.In accepting the mouse group of total length NY-ESO-1 or LVL030, there is not tumour when the off-test from 2 of each group.In accepting the group of LVL068,4 are not had tumour when research finishes.Compare with the mouse of accepting damping fluid, LVL068 and LVL078 give longer survival.Referring to Figure 20.By ELISA, FACS and western blotting assessment NY-ESO-1 specific immune response.By ELISA and FACS assessment LAGE-1a (not containing collagen spline structure territory) specific immune response.Referring to Figure 21.These results are summarized in the following table.

Immunogen	The B16/NY-ESO-1 protection	The NY-ESO-1 specific immunity	The LAGE1a specific immunity
				LVL068	++	++	++
LVL078	+	++	++
				LVL076	+	++	+
LVL024	+	++	+
				LVL030	+	++	+
LVL020	+	+	+
				LVL079	-	+	+
LVL026	-	+	+

Chart 2.Specific immunity is summarized.Keyword: (-)-minimum replying; (+)-medium replying; (++)-the highest replying.

Screening #2

Experimental design.In test in 105 days; whether the CB6F1 mouse is used for assessing each of LVL076, LVL078, LVL068 and LVL024, give at the tumour cell of transplanting with B16/NYESO1 (after 2 immunizations) or with the protection of the subcutaneous attack of B16/LAGE-1a tumour cell (after 4 immunizations) to measure the intramuscular immunization that adds adjuvant with fusion rotein.Particularly, mouse carries out the intramuscular immunization with the 50 μ L injections that comprise 15 μ g protein and 25 μ L AS15 adjuvants.

The group that mouse is divided into 29 mouse/groups.The following immunization of in the time of the 0th, 14,28 and 42 day, carrying out of mouse.

Test

■LVL076

■LVL068

■LVL078

■LVL024

Contrast

■ antigen damping fluid/AS15 damping fluid

■ total length NY-ESO-1

■ does not contain the LAGE-1a in collagen spline structure territory (CLD)

■MAGE A3

10 mouse/groups were attacked with the B16/NY-ESO-1 tumour cell of subcutaneous transplantation in the time of the 28th day.9 mouse/groups were attacked with the B16/LAGE-1A tumour cell of subcutaneous transplantation in the time of the 56th day.In the time of 0,14,28,42 56,84 and 105 day, obtain serum, and by ELISA (IgG1 and IgG2a) assessment for (i) NY-ESO-1 total length, (ii) do not contain the LAGE-1a in collagen spline structure territory and the (iii) antibody response of people's collagen.The experimental design of screening #2 is summarized in Figure 22 and 23.

The result

The B16-NYESO1 tumor challenge.

In accepting the mouse of LVL078, attack the back at B16-NY-ESO-1 for 2 and surpass 50 days no tumours.In the mouse of accepting total length NY-ESO-1 or LVL024, surpass 50 days no tumours from 2 of each group, 3 just work.In accepting the mouse of LVL068,3 no tumours and 4 are just lived.In accepting the mouse of LVL076,3 no tumours and 5 are just lived.Referring to Figure 24.

The B16-LAGE1a tumor challenge.

Accept LVL076 or do not contain all mouse death when attacking back 40 days of the LAGE-1a in collagen sample zone.In the mouse of accepting independent damping fluid, 1 no tumor survival to research finishes.In accepting the mouse of LVL024,1 is not had tumour when research finishes.In the mouse of accepting total length NY-ESO-1, none no tumour, but 1 be still alive when research finishes.In accepting the mouse of LVL078,1 no tumour.In accepting the mouse of LVL068,3 no tumours.In accepting the mouse of LVL076,3 are not had tumour when research finishes.Referring to Figure 25.These results are summarized in down in the chart.

The protection of 3 pairs of B16-LAGE1a tumor challenges of chart.Keyword: (-)-minimum replying; (±)-inferior minimum replying; (+)-medium replying; (++)-the highest replying.

People's collagen specificity immunne response

For whether the collagen spline structure territory of studying NY-ESO-1 stimulates people's collagen specificity immunne response, inoculate back 14 days from mouse and collect serum and merging: (1) damping fluid (contrast) with the one of the following immunization; (2) total length NY-ESO-1; (3) do not contain the LAGE-1a in collagen spline structure territory; (4) LVL068; (5) LVL078; (6) LVL024; (7) LVL076.For in these 7 Serum Banks each, and carry out ELISAs for the positive control that comprises the anti-people's collagen I of mAb.Collagen spline structure territory does not stimulate mouse anti human collagen I antibody to produce.Referring to Figure 26.Carry out similar research (result does not show) for collagen I II and collagen VI; Not detecting mouse anti human collagen I II and mouse anti human collagen VI antibody produces.

Embodiment 4: the purified construct

Use some constructs of listing in the conventional clone technology his-and-hers watches 4 to modify.Particularly, LVL068, LVL030, LVL076, LVL078, LVL024 are modified, to produce LVL155, LVL106, LVL156, LVL157, LVL151.Exist 2 classes to modify, the first kind is the removal of 5 amino-acid residues between 3-protein d and mosaic begin.For example, for LVL024 (SEQ ID NO:74; SEQ ID NO:75) carries out this modification, to produce LVL151 (SEQ ID NO:90; SEQ ID NO:91).Therefore, LVL024 is corresponding to LVL 151.Second type modification is the amino acid whose removal between his-label and mosaic begin.For LVL068 (SEQ ID NO:80; SEQ ID NO:81) carries out this modification, to produce LVL 155 (SEQ ID NO:92; SEQ ID NO:93).Therefore, LVL068 is corresponding to LVL 151.The every kind of fusion protein construct and the corresponding with it fusion protein construct thereof of modifying show in the table 5 of specification sheets.

Should be appreciated that above-described modification do not expect function difference between the fusion rotein that causes fusion rotein and corresponding modification thereof.Therefore, the expection people can utilize and can exchange the fusion rotein of every kind of modification listing on the right side of table 5 with its corresponding fusion rotein of listing on the left-hand side of this chart.

Embodiment 5.

Experimental design.In test in 105 days, the CB6F1 mouse is used for assessing LVL155, LVL106, LVL156, the LVL157 of LVL068, LVL030, LVL076, LVL078, LVL024 and modification, each of LVL151, with research at the tumour cell of transplanting with B16/NYESO1 (after 2 immunizations) or add the intramuscular immunization of adjuvant with fusion rotein with the subcutaneous attack of B16/LAGE-1a tumour cell (after 4 immunizations).Particularly, mouse carries out the intramuscular immunization with the 50 μ L injections that comprise 15 μ g protein and 25 μ L AS15 adjuvants.

The group that mouse is divided into 29 mouse/groups.The following immunization of in the time of the 0th, 14,28 and 42 day, carrying out of mouse.

Test

■LVL068

■LVL030

■LVL076

■LVL078

■LVL024

■LVL155

■LVL106

■LVL156

■LVL157

■LVL151

Contrast

■ antigen damping fluid/AS15 damping fluid

■ total length NY-ESO-1

■ does not contain the LAGE-1a in collagen structure territory

■MAGE A3

10 mouse/groups were attacked with the B16/NY-ESO-1 tumour cell of subcutaneous transplantation in the time of the 28th day.9 mouse/groups were attacked with the B16/LAGE-1A tumour cell of subcutaneous transplantation in the time of the 56th day.In order to monitor specific immune response, can be in the time of 0,14,28,4256,84 and 105 day, obtain serum, and by ELISA (IgG1 and IgG2a) assessment for (i) NY-ESO-1 total length, (ii) do not contain the LAGE-1a in collagen spline structure territory and the (iii) antibody response of people's collagen.

Previous embodiment illustrates for example rather than provides for restriction.

In the application, article " (a) " and " a kind of (an) " are used in reference to one or surpass the grammar object of (being an at least one) article in this article.For example, " element " means one or more elements.As used herein, term " approximately " and " pact " are intended to optional can delete or replaceable with term " accurately " in this article in each case, if by applicant's requirement.

Unit, prefix and symbol can be represented with the generally acknowledged form of its SI.Except as otherwise noted, respectively, nucleic acid with 5 ' from left to right write to 3 ' direction; Aminoacid sequence is from left to right write with amino to carboxyl direction.Digital scope comprises the numeral of limited range.Amino acid can be mentioned by its common known 3 letter characters or by the one-letter symbol of being recommended by IUPAC-IUB biochemical nomenclature commission (Biochemical Nomenclature Commission) in this article.Similarly, Nucleotide can be mentioned by its single-letter code of generally acknowledging usually.Generally speaking term defined above more fully defines with reference to specification sheets.

Sequence table

<110>Denis Martin

Remi Palmantier

＜120〉comprise the fusion rotein of Tumor rejection antigen NY-ESO-1 and LAGE-1

<130>VB62288

<160>98

<170>FastSEQ for Windows Version 4.0

<210>1

<211>867

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>1

atgcaggcgg aaggccgtgg caccggtggt agcaccggcg atgcggatgg tccgggcggt 60

ccgggtattc cggacgggcc tggtggtaat gcgggtgggc caggtgaagc gggtgcgacc 120

ggtggtcgtg gtccgcgggg ggcaggcgca gcacgtgcat ctggtccggg tggtggtgca 180

ccgcgcggtc cgcatggtgg tgcggcgagc ggcctgaatg gttgctgccg ttgcggtgcg 240

cgtggtccgg aaagccgtct gctggaattt tatctggcca tgccgtttgc gaccccgatg 300

gaagcggaac tggcccgtcg tagcctggct caagatgcac cgccgctgcc ggttccgggc 360

gtgctgctga aagaatttac cgtgagcggc aacattctga ccattcgtct gacggcggca 420

gaccatcgtc agctgcaact gagcattagc agctgcctgc aacagctgtc tctgctgatg 480

tggattaccc agtgctttct gccggtgttt ctggcccagc cgccgtctgg tcaacgtggt 540

ggcgcgcgtc gtccggattc tcgcctgctg gaactgcata ttaccatgcc gttcagctct 600

ccaatggagg ccgaattagt gcgtcgcatt ctgagccgtg atgcggcacc gctgccgcgt 660

ccaggtgcgg ttctgaaaga cttcaccgta tctggcaacc tgctgtttat ccgtctgacc 720

gcagcggacc accgccaatt acaattatct atcagctctt gtttacaaca actgtcgctg 780

ttaatgtgga tcactcaatg tttcctgcca gtattcctgg ctcaggcccc gagcggtcag 840

cgtcgtcacc accaccacca ccactaa 867

<210>2

<211>1197

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>2

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa acccaggcgg aaggccgtgg caccggtggt 360

agcaccggcg atgcggatgg tccgggcggt ccgggtattc cggacgggcc tggtggtaat 420

gcgggtgggc caggtgaagc gggtgcgacc ggtggtcgtg gtccgcgggg ggcaggcgca 480

gcacgtgcat ctggtccggg tggtggtgca ccgcgcggtc cgcatggtgg tgcggcgagc 540

ggcctgaatg gttgctgccg ttgcggtgcg cgtggtccgg aaagccgtct gctggaattt 600

tatctggcca tgccgtttgc gaccccgatg gaagcggaac tggcccgtcg tagcctggct 660

caagatgcac cgccgctgcc ggttccgggc gtgctgctga aagaatttac cgtgagcggc 720

aacattctga ccattcgtct gacggcggca gaccatcgtc agctgcaact gagcattagc 780

agctgcctgc aacagctgtc tctgctgatg tggattaccc agtgctttct gccggtgttt 840

ctggcccagc cgccgtctgg tcaacgtggt ggcgcgcgtc gtccggattc tcgcctgctg 900

gaactgcata ttaccatgcc gttcagctct ccaatggagg ccgaattagt gcgtcgcatt 960

ctgagccgtg atgcggcacc gctgccgcgt ccaggtgcgg ttctgaaaga cttcaccgta 1020

tctggcaacc tgctgtttat ccgtctgacc gcagcggacc accgccaatt acaattatct 1080

atcagctctt gtttacaaca actgtcgctg ttaatgtgga tcactcaatg tttcctgcca 1140

gtattcctgg ctcaggcccc gagcggtcag cgtcgtcacc accaccacca ccactaa 1197

<210>3

<211>288

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>3

Met Gln Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp

1 5 10 15

Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly

20 25 30

Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala

35 40 45

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

50 55 60

His Gly Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys Gly Ala

65 70 75 80

Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe

85 90 95

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

100 105 110

Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val

115 120 125

Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln

130 135 140

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

145 150 155 160

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro Ser

165 170 175

Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu

180 185 190

His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg

195 200 205

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

210 215 220

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

225 230 235 240

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

245 250 255

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

260 265 270

Leu Ala Gln Ala Pro Ser Gly Gln Arg Arg His His His His His His

275 280 285

<210>4

<211>398

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>4

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gln

100 105 110

Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp Gly Pro

115 120 125

Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly Gly Pro

130 135 140

Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala Gly Ala

145 150 155 160

Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro His Gly

165 170 175

Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys Gly Ala Arg Gly

180 185 190

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

195 200 205

Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro

210 215 220

Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly

225 230 235 240

Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln

245 250 255

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

260 265 270

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

275 280 285

Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile

290 295 300

Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile

305 310 315 320

Leu Ser Arg Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys

325 330 335

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

340 345 350

Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu

355 360 365

Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala

370 375 380

Gln Ala Pro Ser Gly Gln Arg Arg His His His His His His

385 390 395

<210>5

<211>729

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>5

atggcaggcg cagcacgtgc atctggtccg ggtggtggtg caccgcgcgg tccgcatggt 60

ggtgcggcga gcggcctgaa tggttgctgc cgttgcggtg cgcgtggtcc ggaaagccgt 120

ctgctggaat tttatctggc catgccgttt gcgaccccga tggaagcgga actggcccgt 180

cgtagcctgg ctcaagatgc accgccgctg ccggttccgg gcgtgctgct gaaagaattt 240

accgtgagcg gcaacattct gaccattcgt ctgacggcgg cagaccatcg tcagctgcaa 300

ctgagcatta gcagctgcct gcaacagctg tctctgctga tgtggattac ccagtgcttt 360

ctgccggtgt ttctggccca gccgccgtct ggtcaacgtg gtggcgcgcg tcgtccggat 420

tctcgcctgc tggaactgca tattaccatg ccgttcagct ctccaatgga ggccgaatta 480

gtgcgtcgca ttctgagccg tgatgcggca ccgctgccgc gtccaggtgc ggttctgaaa 540

gacttcaccg tatctggcaa cctgctgttt atccgtctga ccgcagcgga ccaccgccaa 600

ttacaattat ctatcagctc ttgtttacaa caactgtcgc tgttaatgtg gatcactcaa 660

tgtttcctgc cagtattcct ggctcaggcc ccgagcggtc agcgtcgtca ccaccaccac 720

caccactaa 729

<210>6

<211>1059

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>6

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcaggcg cagcacgtgc atctggtccg 360

ggtggtggtg caccgcgcgg tccgcatggt ggtgcggcga gcggcctgaa tggttgctgc 420

cgttgcggtg cgcgtggtcc ggaaagccgt ctgctggaat tttatctggc catgccgttt 480

gcgaccccga tggaagcgga actggcccgt cgtagcctgg ctcaagatgc accgccgctg 540

ccggttccgg gcgtgctgct gaaagaattt accgtgagcg gcaacattct gaccattcgt 600

ctgacggcgg cagaccatcg tcagctgcaa ctgagcatta gcagctgcct gcaacagctg 660

tctctgctga tgtggattac ccagtgcttt ctgccggtgt ttctggccca gccgccgtct 720

ggtcaacgtg gtggcgcgcg tcgtccggat tctcgcctgc tggaactgca tattaccatg 780

ccgttcagct ctccaatgga ggccgaatta gtgcgtcgca ttctgagccg tgatgcggca 840

ccgctgccgc gtccaggtgc ggttctgaaa gacttcaccg tatctggcaa cctgctgttt 900

atccgtctga ccgcagcgga ccaccgccaa ttacaattat ctatcagctc ttgtttacaa 960

caactgtcgc tgttaatgtg gatcactcaa tgtttcctgc cagtattcct ggctcaggcc 1020

ccgagcggtc agcgtcgtca ccaccaccac caccactaa 1059

<210>7

<211>242

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>7

Met Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg

1 5 10 15

Gly Pro His Gly Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys

20 25 30

Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met

35 40 45

Pro Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala

50 55 60

Gln Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe

65 70 75 80

Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His

85 90 95

Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu

100 105 110

Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro

115 120 125

Pro Ser Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu

130 135 140

Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu

145 150 155 160

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

165 170 175

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

180 185 190

Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys

195 200 205

Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro

210 215 220

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

225 230 235 240

His His

<210>8

<211>352

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>8

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

115 120 125

His Gly Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys Gly Ala

130 135 140

Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe

145 150 155 160

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

165 170 175

Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val

180 185 190

Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln

195 200 205

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

210 215 220

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro Ser

225 230 235 240

Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu

245 250 255

His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg

260 265 270

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

275 280 285

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

290 295 300

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

305 310 315 320

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

325 330 335

Leu Ala Gln Ala Pro Ser Gly Gln Arg Arg His His His His His His

340 345 350

<210>9

<211>636

<212>DNA

＜213〉artificial sequence

<220>

＜223〉Ny-ESO-1/LAGE construct

<400>9

atgggtgcgc gtggtccgga aagccgtctg ctggaatttt atctggccat gccgtttgcg 60

accccgatgg aagcggaact ggcccgtcgt agcctggctc aagatgcacc gccgctgccg 120

gttccgggcg tgctgctgaa agaatttacc gtgagcggca acattctgac cattcgtctg 180

acggcggcag accatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 240

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggcccagcc gccgtctggt 300

caacgtggtg gcgcgcgtcg tccggattct cgcctgctgg aactgcatat taccatgccg 360

ttcagctctc caatggaggc cgaattagtg cgtcgcattc tgagccgtga tgcggcaccg 420

ctgccgcgtc caggtgcggt tctgaaagac ttcaccgtat ctggcaacct gctgtttatc 480

cgtctgaccg cagcggacca ccgccaatta caattatcta tcagctcttg tttacaacaa 540

ctgtcgctgt taatgtggat cactcaatgt ttcctgccag tattcctggc tcaggccccg 600

agcggtcagc gtcgtcacca ccaccaccac cactaa 636

<210>10

<211>966

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>10

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accggtgcgc gtggtccgga aagccgtctg 360

ctggaatttt atctggccat gccgtttgcg accccgatgg aagcggaact ggcccgtcgt 420

agcctggctc aagatgcacc gccgctgccg gttccgggcg tgctgctgaa agaatttacc 480

gtgagcggca acattctgac cattcgtctg acggcggcag accatcgtca gctgcaactg 540

agcattagca gctgcctgca acagctgtct ctgctgatgt ggattaccca gtgctttctg 600

ccggtgtttc tggcccagcc gccgtctggt caacgtggtg gcgcgcgtcg tccggattct 660

cgcctgctgg aactgcatat taccatgccg ttcagctctc caatggaggc cgaattagtg 720

cgtcgcattc tgagccgtga tgcggcaccg ctgccgcgtc caggtgcggt tctgaaagac 780

ttcaccgtat ctggcaacct gctgtttatc cgtctgaccg cagcggacca ccgccaatta 840

caattatcta tcagctcttg tttacaacaa ctgtcgctgt taatgtggat cactcaatgt 900

ttcctgccag tattcctggc tcaggccccg agcggtcagc gtcgtcacca ccaccaccac 960

cactaa 966

<210>11

<211>211

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>11

Met Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala

1 5 10 15

Met Pro Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu

20 25 30

Ala Gln Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu

35 40 45

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

50 55 60

His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser

65 70 75 80

Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln

85 90 95

Pro Pro Ser Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu

100 105 110

Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu

115 120 125

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

130 135 140

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

145 150 155 160

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

165 170 175

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

180 185 190

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

195 200 205

His His His

210

<210>12

<211>321

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>12

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gly

100 105 110

Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro

115 120 125

Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln

130 135 140

Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr

145 150 155 160

Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg

165 170 175

Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu

180 185 190

Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro

195 200 205

Ser Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu

210 215 220

Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val

225 230 235 240

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

245 250 255

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

260 265 270

Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu

275 280 285

Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val

290 295 300

Phe Leu Ala Gln Ala Pro Ser Gly Gln Arg Arg His His His His His

305 310 315 320

His

<210>13

<211>870

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>13

atgcaggcgg aaggccgtgg tactggcggt agcaccggcg atgcagatgg tccgggcggt 60

ccgggtattc cggatggtcc gggtggtaat gcaggtggtc caggtgaagc aggtgcgact 120

ggcggtcgtg gtccacgcgg tgcaggtgca gcgcgtgcat ctggtccagg tggcggtgcg 180

ccgcgtggcc cgcatggtgg tgcagctagt gcgcaagatg gtcgttgccc gtgtggtgcg 240

cgtcgtccgg atagccgtct gctggagctg catattacca tgccgtttag cagcccaatg 300

gaagctgagc tggtgcgtcg tattctgtct cgtgacgcag caccgctgcc acgtccgggt 360

gcggttctga aagattttac cgtgagcggc aacctgctgt ttattcgtct gaccgcggca 420

gatcatcgtc agctgcaact gagcattagc agctgcctgc aacagctgtc tctgctgatg 480

tggattaccc agtgctttct gccggtgttt ctggctcagg cgccgtctgg tcagcgtcgt 540

ggtggtgccc gtggcccgga atctcgtctg ctggaatttt atctggccat gccgttcgcg 600

acgccgatgg aagcagagct ggcccgtcgc agcctggctc aggatgcacc gccgctgccg 660

gttccgggcg tgctgctgaa agaatttacg gttagcggta acattctgac catccgtctg 720

accgcagcgg accaccgcca actgcaactg tctatcagct cttgcctgca acaactgtcg 780

ttattaatgt ggatcactca atgtttttta ccagtattcc tggcccaacc gccgagcggc 840

caacgtcgtc accaccacca ccaccactaa 870

<210>14

<211>1200

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>14

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attartgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa acccaggcgg aaggccgtgg tactggcggt 360

agcaccggcg atgcagatgg tccgggcggt ccgggtattc cggatggtcc gggtggtaat 420

gcaggtggtc caggtgaagc aggtgcgact ggcggtcgtg gtccacgcgg tgcaggtgca 480

gcgcgtgcat ctggtccagg tggcggtgcg ccgcgtggcc cgcatggtgg tgcagctagt 540

gcgcaagatg gtcgttgccc gtgtggtgcg cgtcgtccgg atagccgtct gctggagctg 600

catattacca tgccgtttag cagcccaatg gaagctgagc tggtgcgtcg tattctgtct 660

cgtgacgcag caccgctgcc acgtccgggt gcggttctga aagattttac cgtgagcggc 720

aacctgctgt ttattcgtct gaccgcggca gatcatcgtc agctgcaact gagcattagc 780

agctgcctgc aacagctgtc tctgctgatg tggattaccc agtgctttct gccggtgttt 840

ctggctcagg cgccgtctgg tcagcgtcgt ggtggtgccc gtggcccgga atctcgtctg 900

ctggaatttt atctggccat gccgttcgcg acgccgatgg aagcagagct ggcccgtcgc 960

agcctggctc aggatgcacc gccgctgccg gttccgggcg tgctgctgaa agaatttacg 1020

gttagcggta acattctgac catccgtctg accgcagcgg accaccgcca actgcaactg 1080

tctatcagct cttgcctgca acaactgtcg ttattaatgt ggatcactca atgtttttta 1140

ccagtattcc tggcccaacc gccgagcggc caacgtcgtc accaccacca ccaccactaa 1200

<210>15

<211>289

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>15

Met Gln Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp

1 5 10 15

Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly

20 25 30

Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala

35 40 45

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

50 55 60

His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala

65 70 75 80

Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe

85 90 95

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

100 105 110

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

115 120 125

Ser Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln

130 135 140

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

145 150 155 160

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser

165 170 175

Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu

180 185 190

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

195 200 205

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

210 215 220

Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu

225 230 235 240

Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu

245 250 255

Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val

260 265 270

Phe Leu Ala Gln Pro Pro Ser Gly Gln Arg Arg His His His His His

275 280 285

His

<210>16

<211>399

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>16

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gln

100 105 110

Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp Gly Pro

115 120 125

Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly Gly Pro

130 135 140

Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala Gly Ala

145 150 155 160

Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro His Gly

165 170 175

Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala Arg Arg

180 185 190

Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser

195 200 205

Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala Ala

210 215 220

Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser Gly

225 230 235 240

Asn Leu Leu Phe Ile Arg Leu Thr AIa Ala Asp His Arg Gln Leu Gln

245 250 255

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

260 265 270

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

275 280 285

Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr

290 295 300

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

305 310 315 320

Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu

325 330 335

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

340 345 350

Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln

355 360 365

Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu

370 375 380

Ala Gln Pro Pro Ser Gly Gln Arg Arg His His His His His His

385 390 395

<210>17

<211>732

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>17

atggcaggtg cagcgcgtgc atctggtcca ggtggcggtg cgccgcgtgg cccgcatggt 60

ggtgcagcta gtgcgcaaga tggtcgttgc ccgtgtggtg cgcgtcgtcc ggatagccgt 120

ctgctggagc tgcatattac catgccgttt agcagcccaa tggaagctga gctggtgcgt 180

cgtattctgt ctcgtgacgc agcaccgctg ccacgtccgg gtgcggttct gaaagatttt 240

accgtgagcg gcaacctgct gtttattcgt ctgaccgcgg cagatcatcg tcagctgcaa 300

ctgagcatta gcagctgcct gcaacagctg tctctgctga tgtggattac ccagtgcttt 360

ctgccggtgt ttctggctca ggcgccgtct ggtcagcgtc gtggtggtgc ccgtggcccg 420

gaatctcgtc tgctggaatt ttatctggcc atgccgttcg cgacgccgat ggaagcagag 480

ctggcccgtc gcagcctggc tcaggatgca ccgccgctgc cggttccggg cgtgctgctg 540

aaagaattta cggttagcgg taacattctg accatccgtc tgaccgcagc ggaccaccgc 600

caactgcaac tgtctatcag ctcttgcctg caacaactgt cgttattaat gtggatcact 660

caatgttttt taccagtatt cctggcccaa ccgccgagcg gccaacgtcg tcaccaccac 720

caccaccact aa 732

<210>18

<211>1062

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>18

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcaggtg cagcgcgtgc atctggtcca 360

ggtggcggtg cgccgcgtgg cccgcatggt ggtgcagcta gtgcgcaaga tggtcgttgc 420

ccgtgtggtg cgcgtcgtcc ggatagccgt ctgctggagc tgcatattac catgccgttt 480

agcagcccaa tggaagctga gctggtgcgt cgtattctgt ctcgtgacgc agcaccgctg 540

ccacgtccgg gtgcggttct gaaagatttt accgtgagcg gcaacctgct gtttattcgt 600

ctgaccgcgg cagatcatcg tcagctgcaa ctgagcatta gcagctgcct gcaacagctg 660

tctctgctga tgtggattac ccagtgcttt ctgccggtgt ttctggctca ggcgccgtct 720

ggtcagcgtc gtggtggtgc ccgtggcccg gaatctcgtc tgctggaatt ttatctggcc 780

atgccgttcg cgacgccgat ggaagcagag ctggcccgtc gcagcctggc tcaggatgca 840

ccgccgctgc cggttccggg cgtgctgctg aaagaattta cggttagcgg taacattctg 900

accatccgtc tgaccgcagc ggaccaccgc caactgcaac tgtctatcag ctcttgcctg 960

caacaactgt cgttattaat gtggatcact caatgttttt taccagtatt cctggcccaa 1020

ccgccgagcg gccaacgtcg tcaccaccac caccaccact aa 1062

<210>19

<211>243

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>19

Met Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg

1 5 10 15

Gly Pro His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys

20 25 30

Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met

35 40 45

Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser

50 55 60

Arg Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe

65 70 75 80

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

85 90 95

Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu

100 105 110

Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala

115 120 125

Pro Ser Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg Leu

130 135 140

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

145 150 155 160

Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val Pro

165 170 175

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

180 185 190

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

195 200 205

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

210 215 220

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

225 230 235 240

His His His

<210>20

<211>353

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>20

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

115 120 125

His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala

130 135 140

Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe

145 150 155 160

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

165 170 175

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

180 185 190

Ser Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln

195 200 205

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

210 215 220

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser

225 230 235 240

Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu

245 250 255

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

260 265 270

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

275 280 285

Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu

290 295 300

Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu

305 310 315 320

Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val

325 330 335

Phe Leu Ala Gln Pro Pro Ser Gly Gln Arg Arg His His His His His

340 345 350

His

<210>21

<211>639

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>21

atgggtgcgc gtcgtccgga tagccgtctg ctggagctgc atattaccat gccgtttagc 60

agcccaatgg aagctgagct ggtgcgtcgt attctgtctc gtgacgcagc accgctgcca 120

cgtccgggtg cggttctgaa agattttacc gtgagcggca acctgctgtt tattcgtctg 180

accgcggcag atcatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 240

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggctcaggc gccgtctggt 300

cagcgtcgtg gtggtgcccg tggcccggaa tctcgtctgc tggaatttta tctggccatg 360

ccgttcgcga cgccgatgga agcagagctg gcccgtcgca gcctggctca ggatgcaccg 420

ccgctgccgg ttccgggcgt gctgctgaaa gaatttacgg ttagcggtaa cattctgacc 480

atccgtctga ccgcagcgga ccaccgccaa ctgcaactgt ctatcagctc ttgcctgcaa 540

caactgtcgt tattaatgtg gatcactcaa tgttttttac cagtattcct ggcccaaccg 600

ccgagcggcc aacgtcgtca ccaccaccac caccactaa 639

<210>22

<211>969

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>22

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accggtgcgc gtcgtccgga tagccgtctg 360

ctggagctgc atattaccat gccgtttagc agcccaatgg aagctgagct ggtgcgtcgt 420

attctgtctc gtgacgcagc accgctgcca cgtccgggtg cggttctgaa agattttacc 480

gtgagcggca acctgctgtt tattcgtctg accgcggcag atcatcgtca gctgcaactg 540

agcattagca gctgcctgca acagctgtct ctgctgatgt ggattaccca gtgctttctg 600

ccggtgtttc tggctcaggc gccgtctggt cagcgtcgtg gtggtgcccg tggcccggaa 660

tctcgtctgc tggaatttta tctggccatg ccgttcgcga cgccgatgga agcagagctg 720

gcccgtcgca gcctggctca ggatgcaccg ccgctgccgg ttccgggcgt gctgctgaaa 780

gaatttacgg ttagcggtaa cattctgacc atccgtctga ccgcagcgga ccaccgccaa 840

ctgcaactgt ctatcagctc ttgcctgcaa caactgtcgt tattaatgtg gatcactcaa 900

tgttttttac cagtattcct ggcccaaccg ccgagcggcc aacgtcgtca ccaccaccac 960

caccactaa 969

<210>23

<211>212

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>23

Met Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr

1 5 10 15

Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu

20 25 30

Ser Arg Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp

35 40 45

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

50 55 60

His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser

65 70 75 80

Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln

85 90 95

Ala Pro Ser Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg

100 105 110

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

115 120 125

Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val

130 135 140

Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr

145 150 155 160

Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser

165 170 175

Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe

180 185 190

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

195 200 205

His His His His

210

<210>24

<211>322

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>24

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gly

100 105 110

Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro

115 120 125

Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg

130 135 140

Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr

145 150 155 160

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

165 170 175

Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu

180 185 190

Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro

195 200 205

Ser Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu

210 215 220

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

225 230 235 240

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

245 250 255

Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg

260 265 270

Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys

275 280 285

Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro

290 295 300

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

305 310 315 320

His His

<210>25

<211>687

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>25

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgggtgcgcg tggtccggaa agccgtctgc tggaatttta tctggccatg 120

ccgtttgcga ccccgatgga agcggaactg gcccgtcgta gcctggctca agatgcaccg 180

ccgctgccgg ttccgggcgt gctgctgaaa gaatttaccg tgagcggcaa cattctgacc 240

attcgtctga cggcggcaga ccatcgtcag ctgcaactga gcattagcag ctgcctgcaa 300

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggcccagccg 360

ccgtctggtc aacgtggtgg cgcgcgtcgt ccggattctc gcctgctgga actgcatatt 420

accatgccgt tcagctctcc aatggaggcc gaattagtgc gtcgcattct gagccgtgat 480

gcggcaccgc tgccgcgtcc aggtgcggtt ctgaaagact tcaccgtatc tggcaacctg 540

ctgtttatcc gtctgaccgc agcggaccac cgccaattac aattatctat cagctcttgt 600

ttacaacaac tgtcgctgtt aatgtggatc actcaatgtt tcctgccagt attcctggct 660

caggccccga gcggtcagcg tcgttaa 687

<210>26

<211>228

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>26

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gly Ala Arg Gly Pro Glu Ser Arg

20 25 30

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

35 40 45

Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val

50 55 60

Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr

65 70 75 80

Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser

85 90 95

Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe

100 105 110

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

115 120 125

Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe

130 135 140

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

145 150 155 160

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

165 170 175

Ser Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln

180 185 190

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

195 200 205

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser

210 215 220

Gly Gln Arg Arg

225

<210>27

<211>636

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>27

atgcatcatc atcatcatca cggtgcgcgt ggtccggaaa gccgtctgct ggaattttat 60

ctggccatgc cgtttgcgac cccgatggaa gcggaactgg cccgtcgtag cctggctcaa 120

gatgcaccgc cgctgccggt tccgggcgtg ctgctgaaag aatttaccgt gagcggcaac 180

attctgacca ttcgtctgac ggcggcagac catcgtcagc tgcaactgag cattagcagc 240

tgcctgcaac agctgtctct gctgatgtgg attacccagt gctttctgcc ggtgtttctg 300

gcccagccgc cgtctggtca acgtggtggc gcgcgtcgtc cggattctcg cctgctggaa 360

ctgcatatta ccatgccgtt cagctctcca atggaggccg aattagtgcg tcgcattctg 420

agccgtgatg cggcaccgct gccgcgtcca ggtgcggttc tgaaagactt caccgtatct 480

ggcaacctgc tgtttatccg tctgaccgca gcggaccacc gccaattaca attatctatc 540

agctcttgtt tacaacaact gtcgctgtta atgtggatca ctcaatgttt cctgccagta 600

ttcctggctc aggccccgag cggtcagcgt cgttaa 636

<210>28

<211>211

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ES01/LAGE construct

<400>28

Met His His His His His His Gly Ala Arg Gly Pro Glu Ser Arg Leu

1 5 10 15

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

20 25 30

Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val Pro

35 40 45

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

50 55 60

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

65 70 75 80

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

85 90 95

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

100 105 110

Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser

115 120 125

Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala

130 135 140

Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser

145 150 155 160

Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu

165 170 175

Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp

180 185 190

Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser Gly

195 200 205

Gln Arg Arg

210

<210>29

<211>780

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>29

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tggcaggcgc agcacgtgca tctggtccgg gtggtggtgc accgcgcggt 120

ccgcatggtg gtgcggcgag cggcctgaat ggttgctgcc gttgcggtgc gcgtggtccg 180

gaaagccgtc tgctggaatt ttatctggcc atgccgtttg cgaccccgat ggaagcggaa 240

ctggcccgtc gtagcctggc tcaagatgca ccgccgctgc cggttccggg cgtgctgctg 300

aaagaattta ccgtgagcgg caacattctg accattcgtc tgacggcggc agaccatcgt 360

cagctgcaac tgagcattag cagctgcctg caacagctgt ctctgctgat gtggattacc 420

cagtgctttc tgccggtgtt tctggcccag ccgccgtctg gtcaacgtgg tggcgcgcgt 480

cgtccggatt ctcgcctgct ggaactgcat attaccatgc cgttcagctc tccaatggag 540

gccgaattag tgcgtcgcat tctgagccgt gatgcggcac cgctgccgcg tccaggtgcg 600

gttctgaaag acttcaccgt atctggcaac ctgctgttta tccgtctgac cgcagcggac 660

caccgccaat tacaattatc tatcagctct tgtttacaac aactgtcgct gttaatgtgg 720

atcactcaat gtttcctgcc agtattcctg gctcaggccc cgagcggtca gcgtcgttaa 780

<210>30

<211>259

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>30

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Ala Gly Ala Ala Arg Ala Ser Gly

20 25 30

Pro Gly Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Gly

35 40 45

Leu Asn Gly Cys Cys Arg Cys Gly Ala Arg Gly Pro Glu Ser Arg Leu

50 55 60

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

65 70 75 80

Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val Pro

85 90 95

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

100 105 110

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

115 120 125

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

130 135 140

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

145 150 155 160

Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser

165 170 175

Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala

180 185 190

Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser

195 200 205

Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu

210 215 220

Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp

225 230 235 240

Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser Gly

245 250 255

Gln Arg Arg

<210>31

<211>729

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>31

atgcatcatc atcatcatca cgcaggcgca gcacgtgcat ctggtccggg tggtggtgca 60

ccgcgcggtc cgcatggtgg tgcggcgagc ggcctgaatg gttgctgccg ttgcggtgcg 120

cgtggtccgg aaagccgtct gctggaattt tatctggcca tgccgtttgc gaccccgatg 180

gaagcggaac tggcccgtcg tagcctggct caagatgcac cgccgctgcc ggttccgggc 240

gtgctgctga aagaatttac cgtgagcggc aacattctga ccattcgtct gacggcggca 300

gaccatcgtc agctgcaact gagcattagc agctgcctgc aacagctgtc tctgctgatg 360

tggattaccc agtgctttct gccggtgttt ctggcccagc cgccgtctgg tcaacgtggt 420

ggcgcgcgtc gtccggattc tcgcctgctg gaactgcata ttaccatgcc gttcagctct 480

ccaatggagg ccgaattagt gcgtcgcatt ctgagccgtg atgcggcacc gctgccgcgt 540

ccaggtgcgg ttctgaaaga cttcaccgta tctggcaacc tgctgtttat ccgtctgacc 600

gcagcggacc accgccaatt acaattatct atcagctctt gtttacaaca actgtcgctg 660

ttaatgtgga tcactcaatg tttcctgcca gtattcctgg ctcaggcccc gagcggtcag 720

cgtcgttaa 729

<210>32

<211>242

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>32

Met His His His His His His Ala Gly Ala Ala Arg Ala Ser Gly Pro

1 5 10 15

Gly Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Gly Leu

20 25 30

Asn Gly Cys Cys Arg Cys Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu

35 40 45

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

50 55 60

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

65 70 75 80

Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg

85 90 95

Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys

100 105 110

Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro

115 120 125

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

130 135 140

Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser

145 150 155 160

Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala Ala

165 170 175

Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser Gly

180 185 190

Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln

195 200 205

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

210 215 220

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

225 230 235 240

Arg Arg

<210>33

<211>918

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>33

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgcaggcgga aggccgtggc accggtggta gcaccggcga tgcggatggt 120

ccgggcggtc cgggtattcc ggacgggcct ggtggtaatg cgggtgggcc aggtgaagcg 180

ggtgcgaccg gtggtcgtgg tccgcggggg gcaggcgcag cacgtgcatc tggtccgggt 240

ggtggtgcac cgcgcggtcc gcatggtggt gcggcgagcg gcctgaatgg ttgctgccgt 300

tgcggtgcgc gtggtccgga aagccgtctg ctggaatttt atctggccat gccgtttgcg 360

accccgatgg aagcggaact ggcccgtcgt agcctggctc aagatgcacc gccgctgccg 420

gttccgggcg tgctgctgaa agaatttacc gtgagcggca acattctgac cattcgtctg 480

acggcggcag accatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 540

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggcccagcc gccgtctggt 600

caacgtggtg gcgcgcgtcg tccggattct cgcctgctgg aactgcatat taccatgccg 660

ttcagctctc caatggaggc cgaattagtg cgtcgcattc tgagccgtga tgcggcaccg 720

ctgccgcgtc caggtgcggt tctgaaagac ttcaccgtat ctggcaacct gctgtttatc 780

cgtctgaccg cagcggacca ccgccaatta caattatcta tcagctcttg tttacaacaa 840

ctgtcgctgt taatgtggat cactcaatgt ttcctgccag tattcctggc tcaggccccg 900

agcggtcagc gtcgttaa 918

<210>34

<211>305

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>34

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gln Ala Glu Gly Arg Gly Thr Gly

20 25 30

Gly Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp

35 40 45

Gly Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly

50 55 60

Gly Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly

65 70 75 80

Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Gly Leu Asn

85 90 95

Gly Cys Cys Arg Cys Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu

100 105 110

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

115 120 125

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

130 135 140

Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu

145 150 155 160

Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu

165 170 175

Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val

180 185 190

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

195 200 205

Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro

210 215 220

Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala Ala Pro

225 230 235 240

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

245 250 255

Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu

260 265 270

Ser Ile Ser Ser Cys Leu Gln Gln Leu Set Leu Leu Met Trp Ile Thr

275 280 285

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

290 295 300

Arg

305

<210>35

<211>867

<212>DNA

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>35

atgcatcatc atcatcatca ccaggcggaa ggccgtggca ccggtggtag caccggcgat 60

gcggatggtc cgggcggtcc gggtattccg gacgggcctg gtggtaatgc gggtgggcca 120

ggtgaagcgg gtgcgaccgg tggtcgtggt ccgcgggggg caggcgcagc acgtgcatct 180

ggtccgggtg gtggtgcacc gcgcggtccg catggtggtg cggcgagcgg cctgaatggt 240

tgctgccgtt gcggtgcgcg tggtccggaa agccgtctgc tggaatttta tctggccatg 300

ccgtttgcga ccccgatgga agcggaactg gcccgtcgta gcctggctca agatgcaccg 360

ccgctgccgg ttccgggcgt gctgctgaaa gaatttaccg tgagcggcaa cattctgacc 420

attcgtctga cggcggcaga ccatcgtcag ctgcaactga gcattagcag ctgcctgcaa 480

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggcccagccg 540

ccgtctggtc aacgtggtgg cgcgcgtcgt ccggattctc gcctgctgga actgcatatt 600

accatgccgt tcagctctcc aatggaggcc gaattagtgc gtcgcattct gagccgtgat 660

gcggcaccgc tgccgcgtcc aggtgcggtt ctgaaagact tcaccgtatc tggcaacctg 720

ctgtttatcc gtctgaccgc agcggaccac cgccaattac aattatctat cagctcttgt 780

ttacaacaac tgtcgctgtt aatgtggatc actcaatgtt tcctgccagt attcctggct 840

caggccccga gcggtcagcg tcgttaa 867

<210>36

<211>288

<212>PRT

＜213〉artificial sequence

<220>

＜223〉NY-ESO-1/LAGE construct

<400>36

Met His His His His His His Gln Ala Glu Gly Arg Gly Thr Gly Gly

1 5 10 15

Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly

20 25 30

Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly

35 40 45

Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly

50 55 60

Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Gly Leu Asn Gly

65 70 75 80

Cys Cys Arg Cys Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe

85 90 95

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

100 105 110

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

115 120 125

Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr

130 135 140

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

145 150 155 160

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

165 170 175

Leu Ala Gln Pro Pro Ser Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp

180 185 190

Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met

195 200 205

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

210 215 220

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

225 230 235 240

Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser

245 250 255

Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln

260 265 270

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

275 280 285

<210>37

<211>690

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>37

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgggtgcgcg tcgtccggat agccgtctgc tggagctgca tattaccatg 120

ccgtttagca gcccaatgga agctgagctg gtgcgtcgta ttctgtctcg tgacgcagca 180

ccgctgccac gtccgggtgc ggttctgaaa gattttaccg tgagcggcaa cctgctgttt 240

attcgtctga ccgcggcaga tcatcgtcag ctgcaactga gcattagcag ctgcctgcaa 300

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggctcaggcg 360

ccgtctggtc agcgtcgtgg tggtgcccgt ggcccggaat ctcgtctgct ggaattttat 420

ctggccatgc cgttcgcgac gccgatggaa gcagagctgg cccgtcgcag cctggctcag 480

gatgcaccgc cgctgccggt tccgggcgtg ctgctgaaag aatttacggt tagcggtaac 540

attctgacca tccgtctgac cgcagcggac caccgccaac tgcaactgtc tatcagctct 600

tgcctgcaac aactgtcgtt attaatgtgg atcactcaat gttttttacc agtattcctg 660

gcccaaccgc cgagcggcca acgtcgttaa 690

<210>38

<211>229

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>38

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gly Ala Arg Arg Pro Asp Ser Arg

20 25 30

Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala

35 40 45

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

50 55 60

Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser Gly Asn Leu Leu Phe

65 70 75 80

Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser

85 90 95

Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe

100 105 110

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

115 120 125

Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro

130 135 140

Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln

145 150 155 160

Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr

165 170 175

Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg

180 185 190

Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu

195 200 205

Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro

210 215 220

Ser Gly Gln Arg Arg

225

<210>39

<211>639

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>39

atgcatcatc atcatcatca cggtgcgcgt cgtccggata gccgtctgct ggagctgcat 60

attaccatgc cgtttagcag cccaatggaa gctgagctgg tgcgtcgtat tctgtctcgt 120

gacgcagcac cgctgccacg tccgggtgcg gttctgaaag attttaccgt gagcggcaac 180

ctgctgttta ttcgtctgac cgcggcagat catcgtcagc tgcaactgag cattagcagc 240

tgcctgcaac agctgtctct gctgatgtgg attacccagt gctttctgcc ggtgtttctg 300

gctcaggcgc cgtctggtca gcgtcgtggt ggtgcccgtg gcccggaatc tcgtctgctg 360

gaattttatc tggccatgcc gttcgcgacg ccgatggaag cagagctggc ccgtcgcagc 420

ctggctcagg atgcaccgcc gctgccggtt ccgggcgtgc tgctgaaaga atttacggtt 480

agcggtaaca ttctgaccat ccgtctgacc gcagcggacc accgccaact gcaactgtct 540

atcagctctt gcctgcaaca actgtcgtta ttaatgtgga tcactcaatg ttttttacca 600

gtattcctgg cccaaccgcc gagcggccaa cgtcgttaa 639

<210>40

<211>212

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>40

Met His His His His His His Gly Ala Arg Arg Pro Asp Ser Arg Leu

1 5 10 15

Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu

20 25 30

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

35 40 45

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

50 55 60

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

65 70 75 80

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

85 90 95

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

100 105 110

Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe

115 120 125

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

130 135 140

Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val

145 150 155 160

Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln

165 170 175

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

180 185 190

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro Ser

195 200 205

Gly Gln Arg Arg

210

<210>41

<211>783

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>41

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tggcaggtgc agcgcgtgca tctggtccag gtggcggtgc gccgcgtggc 120

ccgcatggtg gtgcagctag tgcgcaagat ggtcgttgcc cgtgtggtgc gcgtcgtccg 180

gatagccgtc tgctggagct gcatattacc atgccgttta gcagcccaat ggaagctgag 240

ctggtgcgtc gtattctgtc tcgtgacgca gcaccgctgc cacgtccggg tgcggttctg 300

aaagatttta ccgtgagcgg caacctgctg tttattcgtc tgaccgcggc agatcatcgt 360

cagctgcaac tgagcattag cagctgcctg caacagctgt ctctgctgat gtggattacc 420

cagtgctttc tgccggtgtt tctggctcag gcgccgtctg gtcagcgtcg tggtggtgcc 480

cgtggcccgg aatctcgtct gctggaattt tatctggcca tgccgttcgc gacgccgatg 540

gaagcagagc tggcccgtcg cagcctggct caggatgcac cgccgctgcc ggttccgggc 600

gtgctgctga aagaatttac ggttagcggt aacattctga ccatccgtct gaccgcagcg 660

gaccaccgcc aactgcaact gtctatcagc tcttgcctgc aacaactgtc gttattaatg 720

tggatcactc aatgtttttt accagtattc ctggcccaac cgccgagcgg ccaacgtcgt 780

taa 783

<210>42

<211>260

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>42

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Ala Gly Ala Ala Arg Ala Ser Gly

20 25 30

Pro Gly Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Ala

35 40 45

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

50 55 60

Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu

65 70 75 80

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

85 90 95

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

100 105 110

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

115 120 125

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

130 135 140

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

145 150 155 160

Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe

165 170 175

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

180 185 190

Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val

195 200 205

Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln

210 215 220

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

225 230 235 240

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro Ser

245 250 255

Gly Gln Arg Arg

260

<210>43

<211>732

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>43

atgcatcatc atcatcatca cgcaggtgca gcgcgtgcat ctggtccagg tggcggtgcg 60

ccgcgtggcc cgcatggtgg tgcagctagt gcgcaagatg gtcgttgccc gtgtggtgcg 120

cgtcgtccgg atagccgtct gctggagctg catattacca tgccgtttag cagcccaatg 180

gaagctgagc tggtgcgtcg tattctgtct cgtgacgcag caccgctgcc acgtccgggt 240

gcggttctga aagattttac cgtgagcggc aacctgctgt ttattcgtct gaccgcggca 300

gatcatcgtc agctgcaact gagcattagc agctgcctgc aacagctgtc tctgctgatg 360

tggattaccc agtgctttct gccggtgttt ctggctcagg cgccgtctgg tcagcgtcgt 420

ggtggtgccc gtggcccgga atctcgtctg ctggaatttt atctggccat gccgttcgcg 480

acgccgatgg aagcagagct ggcccgtcgc agcctggctc aggatgcacc gccgctgccg 540

gttccgggcg tgctgctgaa agaatttacg gttagcggta acattctgac catccgtctg 600

accgcagcgg accaccgcca actgcaactg tctatcagct cttgcctgca acaactgtcg 660

ttattaatgt ggatcactca atgtttttta ccagtattcc tggcccaacc gccgagcggc 720

caacgtcgtt aa 732

<210>44

<211>243

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>44

Met His His His His His His Ala Gly Ala Ala Arg Ala Ser Gly Pro

1 5 10 15

Gly Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Ala Gln

20 25 30

Asp Gly Arg Cys Pro Cys Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu

35 40 45

Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu

50 55 60

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

65 70 75 80

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

85 90 95

Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys

100 105 110

Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro

115 120 125

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

130 135 140

Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe Ala

145 150 155 160

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

165 170 175

Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser

180 185 190

Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu

195 200 205

Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp

210 215 220

Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro Ser Gly

225 230 235 240

Gln Arg Arg

<210>45

<211>921

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>45

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgcaggcgga aggccgtggt actggcggta gcaccggcga tgcagatggt 120

ccgggcggtc cgggtattcc ggatggtccg ggtggtaatg caggtggtcc aggtgaagca 180

ggtgcgactg gcggtcgtgg tccacgcggt gcaggtgcag cgcgtgcatc tggtccaggt 240

ggcggtgcgc cgcgtggccc gcatggtggt gcagctagtg cgcaagatgg tcgttgcccg 300

tgtggtgcgc gtcgtccgga tagccgtctg ctggagctgc atattaccat gccgtttagc 360

agcccaatgg aagctgagct ggtgcgtcgt attctgtctc gtgacgcagc accgctgcca 420

cgtccgggtg cggttctgaa agattttacc gtgagcggca acctgctgtt tattcgtctg 480

accgcggcag atcatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 540

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggctcaggc gccgtctggt 600

cagcgtcgtg gtggtgcccg tggcccggaa tctcgtctgc tggaatttta tctggccatg 660

ccgttcgcga cgccgatgga agcagagctg gcccgtcgca gcctggctca ggatgcaccg 720

ccgctgccgg ttccgggcgt gctgctgaaa gaatttacgg ttagcggtaa cattctgacc 780

atccgtctga ccgcagcgga ccaccgccaa ctgcaactgt ctatcagctc ttgcctgcaa 840

caactgtcgt tattaatgtg gatcactcaa tgttttttac cagtattcct ggcccaaccg 900

ccgagcggcc aacgtcgtta a 921

<210>46

<211>306

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>46

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gln Ala Glu Gly Arg Gly Thr Gly

20 25 30

Gly Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp

35 40 45

Gly Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly

50 55 60

Gly Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly

65 70 75 80

Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Ala Gln Asp

85 90 95

Gly Arg Cys Pro Cys Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu

100 105 110

Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val

115 120 125

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

130 135 140

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

145 150 155 160

Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu

165 170 175

Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val

180 185 190

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

195 200 205

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

210 215 220

Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro

225 230 235 240

Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly

245 250 255

Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln

260 265 270

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

275 280 285

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

290 295 300

Arg Arg

305

<210>47

<211>870

<212>DNA

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>47

atgcatcatc atcatcatca ccaggcggaa ggccgtggta ctggcggtag caccggcgat 60

gcagatggtc cgggcggtcc gggtattccg gatggtccgg gtggtaatgc aggtggtcca 120

ggtgaagcag gtgcgactgg cggtcgtggt ccacgcggtg caggtgcagc gcgtgcatct 180

ggtccaggtg gcggtgcgcc gcgtggcccg catggtggtg cagctagtgc gcaagatggt 240

cgttgcccgt gtggtgcgcg tcgtccggat agccgtctgc tggagctgca tattaccatg 300

ccgtttagca gcccaatgga agctgagctg gtgcgtcgta ttctgtctcg tgacgcagca 360

ccgctgccac gtccgggtgc ggttctgaaa gattttaccg tgagcggcaa cctgctgttt 420

attcgtctga ccgcggcaga tcatcgtcag ctgcaactga gcattagcag ctgcctgcaa 480

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggctcaggcg 540

ccgtctggtc agcgtcgtgg tggtgcccgt ggcccggaat ctcgtctgct ggaattttat 600

ctggccatgc cgttcgcgac gccgatggaa gcagagctgg cccgtcgcag cctggctcag 660

gatgcaccgc cgctgccggt tccgggcgtg ctgctgaaag aatttacggt tagcggtaac 720

attctgacca tccgtctgac cgcagcggac caccgccaac tgcaactgtc tatcagctct 780

tgcctgcaac aactgtcgtt attaatgtgg atcactcaat gttttttacc agtattcctg 840

gcccaaccgc cgagcggcca acgtcgttaa 870

<210>48

<211>289

<212>PRT

＜213〉artificial sequence

<220>

＜223〉LAGE/NY-ESO-1 construct

<400>48

Met His His His His His His Gln Ala Glu Gly Arg Gly Thr Gly Gly

1 5 10 15

Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly

20 25 30

Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly

35 40 45

Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly

50 55 60

Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Ala Gln Asp Gly

65 70 75 80

Arg Cys Pro Cys Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu

85 90 95

His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg

100 105 110

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

115 120 125

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

130 135 140

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

145 150 155 160

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

165 170 175

Leu Ala Gln Ala Pro Ser Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro

180 185 190

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

195 200 205

Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro

210 215 220

Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn

225 230 235 240

Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu

245 250 255

Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr

260 265 270

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

275 280 285

Arg

<210>49

<211>180

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>49

Met Gln Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp

1 5 10 15

Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly

20 25 30

Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala

35 40 45

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

50 55 60

His Gly Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys Gly Ala

65 70 75 80

Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe

85 90 95

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

100 105 110

Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val

115 120 125

Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln

130 135 140

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

145 150 155 160

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro Ser

165 170 175

Gly Gln Arg Arg

180

<210>50

<211>420

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>50

atggcaggcg cagcacgtgc atctggtccg ggtggtggtg caccgcgcgg tccgcatggt 60

ggtgcggcga gcggcctgaa tggttgctgc cgttgcggtg cgcgtggtcc ggaaagccgt 120

ctgctggaat tttatctggc catgccgttt gcgaccccga tggaagcgga actggcccgt 180

cgtagcctgg ctcaagatgc accgccgctg ccggttccgg gcgtgctgct gaaagaattt 240

accgtgagcg gcaacattct gaccattcgt ctgacggcgg cagaccatcg tcagctgcaa 300

ctgagcatta gcagctgcct gcaacagctg tctctgctga tgtggattac ccagtgcttt 360

ctgccggtgt ttctggccca gccgccgtct ggtcaacgtc accaccacca ccaccactaa 420

<210>51

<211>139

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>51

Met Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg

1 5 10 15

Gly Pro His Gly Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys

20 25 30

Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met

35 40 45

Pro Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala

50 55 60

Gln Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe

65 70 75 80

Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His

85 90 95

Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu

100 105 110

Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro

115 120 125

Pro Ser Gly Gln Arg His His His His His His

130 135

<210>52

<211>750

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>52

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcaggcg cagcacgtgc atctggtccg 360

ggtggtggtg caccgcgcgg tccgcatggt ggtgcggcga gcggcctgaa tggttgctgc 420

cgttgcggtg cgcgtggtcc ggaaagccgt ctgctggaat tttatctggc catgccgttt 480

gcgaccccga tggaagcgga actggcccgt cgtagcctgg ctcaagatgc accgccgctg 540

ccggttccgg gcgtgctgct gaaagaattt accgtgagcg gcaacattct gaccattcgt 600

ctgacggcgg cagaccatcg tcagctgcaa ctgagcatta gcagctgcct gcaacagctg 660

tctctgctga tgtggattac ccagtgcttt ctgccggtgt ttctggccca gccgccgtct 720

ggtcaacgtc accaccacca ccaccactaa 750

<210>53

<211>249

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>53

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

115 120 125

His Gly Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys Gly Ala

130 135 140

Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro Phe

145 150 155 160

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

165 170 175

Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val

180 185 190

Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln

195 200 205

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

210 215 220

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro Ser

225 230 235 240

Gly Gln Arg His His His His His His

245

<210>54

<211>327

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>54

atgggtgcgc gtggtccgga aagccgtctg ctggaatttt atctggccat gccgtttgcg 60

accccgatgg aagcggaact ggcccgtcgt agcctggctc aagatgcacc gccgctgccg 120

gttccgggcg tgctgctgaa agaatttacc gtgagcggca acattctgac cattcgtctg 180

acggcggcag accatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 240

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggcccagcc gccgtctggt 300

caacgtcacc accaccacca ccactaa 327

<210>55

<211>108

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>55

Met Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala

1 5 10 15

Met Pro Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu

20 25 30

Ala Gln Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu

35 40 45

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

50 55 60

His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser

65 70 75 80

Leu Leu Met Trp Ile Thr Gln Cys Phc Leu Pro Val Phe Leu Ala Gln

85 90 95

Pro Pro Ser Gly Gln Arg His His His His His His

100 105

<210>56

<211>657

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>56

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accggtgcgc gtggtccgga aagccgtctg 360

ctggaatttt atctggccat gccgtttgcg accccgatgg aagcggaact ggcccgtcgt 420

agcctggctc aagatgcacc gccgctgccg gttccgggcg tgctgctgaa agaatttacc 480

gtgagcggca acattctgac cattcgtctg acggcggcag accatcgtca gctgcaactg 540

agcattagca gctgcctgca acagctgtct ctgctgatgt ggattaccca gtgctttctg 600

ccggtgtttc tggcccagcc gccgtctggt caacgtcacc accaccacca ccactaa 657

<210>57

<211>218

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>57

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gly

100 105 110

Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro

115 120 125

Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln

130 135 140

Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr

145 150 155 160

Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg

165 170 175

Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu

180 185 190

Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro

195 200 205

Ser Gly Gln Arg His His His His His His

210 215

<210>58

<211>180

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<220>

<221>PEPTIDE

<222>(0)...(0)

<223>Lage 1a

<400>58

Met Gln Ala Glu Gly Gln Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp

1 5 10 15

Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly

20 25 30

Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala

35 40 45

Gly Ala Ala Arg Ala Ser Gly Pro Arg Gly Gly Ala Pro Arg Gly Pro

50 55 60

His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala

65 70 75 80

Arg Arg Pro Asp Ser Arg Leu Leu Gln Leu His Ile Thr Met Pro Phe

85 90 95

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

100 105 110

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

115 120 125

Ser Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln

130 135 140

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

145 150 155 160

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser

165 170 175

Gly Gln Arg Arg

180

<210>59

<211>561

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>59

atgcaggcgg aaggccgtgg tactggcggt agcaccggcg atgcagatgg tccgggcggt 60

ccgggtattc cggatggtcc gggtggtaat gcaggtggtc caggtgaagc aggtgcgact 120

ggcggtcgtg gtccacgcgg tgcaggtgca gcgcgtgcat ctggtccagg tggcggtgcg 180

ccgcgtggcc cgcatggtgg tgcagctagt gcgcaagatg gtcgttgccc gtgtggtgcg 240

cgtcgtccgg atagccgtct gctggagctg catattacca tgccgtttag cagcccaatg 300

gaagctgagc tggtgcgtcg tattctgtct cgtgacgcag caccgctgcc acgtccgggt 360

gcggttctga aagattttac cgtgagcggc aacctgctgt ttattcgtct gaccgcggca 420

gatcatcgtc agctgcaact gagcattagc agctgcctgc aacagctgtc tctgctgatg 480

tggattaccc agtgctttct gccggtgttt ctggctcagg cgccgtctgg tcagcgtcgt 540

caccaccacc accaccacta a 561

<210>60

<211>186

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>60

Met Gln Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp

1 5 10 15

Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly

20 25 30

Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala

35 40 45

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

50 55 60

His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala

65 70 75 80

Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe

85 90 95

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

100 105 110

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

115 120 125

Ser Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln

130 135 140

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

145 150 155 160

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser

165 170 175

Gly Gln Arg Arg His His His His His His

180 185

<210>61

<211>891

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>61

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa acccaggcgg aaggccgtgg tactggcggt 360

agcaccggcg atgcagatgg tccgggcggt ccgggtattc cggatggtcc gggtggtaat 420

gcaggtggtc caggtgaagc aggtgcgact ggcggtcgtg gtccacgcgg tgcaggtgca 480

gcgcgtgcat ctggtccagg tggcggtgcg ccgcgtggcc cgcatggtgg tgcagctagt 540

gcgcaagatg gtcgttgccc gtgtggtgcg cgtcgtccgg atagccgtct gctggagctg 600

catattacca tgccgtttag cagcccaatg gaagctgagc tggtgcgtcg tattctgtct 660

cgtgacgcag caccgctgcc acgtccgggt gcggttctga aagattttac cgtgagcggc 720

aacctgctgt ttattcgtct gaccgcggca gatcatcgtc agctgcaact gagcattagc 780

agctgcctgc aacagctgtc tctgctgatg tggattaccc agtgctttct gccggtgttt 840

ctggctcagg cgccgtctgg tcagcgtcgt caccaccacc accaccacta a 891

<210>62

<211>296

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>62

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gln

100 105 110

Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp Gly Pro

115 120 125

Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly Gly Pro

130 135 140

Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala Gly Ala

145 150 155 160

Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro His Gly

165 170 175

Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala Arg Arg

180 185 190

Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser

195 200 205

Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala Ala

210 215 220

Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser Gly

225 230 235 240

Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln

245 250 255

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

260 265 270

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

275 280 285

Arg Arg His His His His His His

290 295

<210>63

<211>423

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>63

atggcaggtg cagcgcgtgc atctggtcca ggtggcggtg cgccgcgtgg cccgcatggt 60

ggtgcagcta gtgcgcaaga tggtcgttgc ccgtgtggtg cgcgtcgtcc ggatagccgt 120

ctgctggagc tgcatattac catgccgttt agcagcccaa tggaagctga gctggtgcgt 180

cgtattctgt ctcgtgacgc agcaccgctg ccacgtccgg gtgcggttct gaaagatttt 240

accgtgagcg gcaacctgct gtttattcgt ctgaccgcgg cagatcatcg tcagctgcaa 300

ctgagcatta gcagctgcct gcaacagctg tctctgctga tgtggattac ccagtgcttt 360

ctgccggtgt ttctggctca ggcgccgtct ggtcagcgtc gtcaccacca ccaccaccac 420

taa 423

<210>64

<211>140

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>64

Met Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg

1 5 10 15

Gly Pro His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys

20 25 30

Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met

35 40 45

Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser

50 55 60

Arg Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe

65 70 75 80

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

85 90 95

Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu

100 105 110

Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala

115 120 125

Pro Ser Gly Gln Arg Arg His His His His His His

130 135 140

<210>65

<211>753

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>65

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcaggtg cagcgcgtgc atctggtcca 360

ggtggcggtg cgccgcgtgg cccgcatggt ggtgcagcta gtgcgcaaga tggtcgttgc 420

ccgtgtggtg cgcgtcgtcc ggatagccgt ctgctggagc tgcatattac catgccgttt 480

agcagcccaa tggaagctga gctggtgcgt cgtattctgt ctcgtgacgc agcaccgctg 540

ccacgtccgg gtgcggttct gaaagatttt accgtgagcg gcaacctgct gtttattcgt 600

ctgaccgcgg cagatcatcg tcagctgcaa ctgagcatta gcagctgcct gcaacagctg 660

tctctgctga tgtggattac ccagtgcttt ctgccggtgt ttctggctca ggcgccgtct 720

ggtcagcgtc gtcaccacca ccaccaccac taa 753

<210>66

<211>250

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>66

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro

115 120 125

His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala

130 135 140

Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe

145 150 155 160

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

165 170 175

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

180 185 190

Ser Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln

195 200 205

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

210 215 220

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser

225 230 235 240

Gly Gln Arg Arg His His His His His His

245 250

<210>67

<211>330

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>67

atgggtgcgc gtcgtccgga tagccgtctg ctggagctgc atattaccat gccgtttagc 60

agcccaatgg aagctgagct ggtgcgtcgt attctgtctc gtgacgcagc accgctgcca 120

cgtccgggtg cggttctgaa agattttacc gtgagcggca acctgctgtt tattcgtctg 180

accgcggcag atcatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 240

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggctcaggc gccgtctggt 300

cagcgtcgtc accaccacca ccaccactaa 330

<210>68

<211>109

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>68

Met Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr

1 5 10 15

Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu

20 25 30

Ser Arg Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp

35 40 45

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

50 55 60

His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser

65 70 75 80

Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln

85 90 95

Ala Pro Ser Gly Gln Arg Arg His His His His His His

100 105

<210>69

<211>660

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>69

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accggtgcgc gtcgtccgga tagccgtctg 360

ctggagctgc atattaccat gccgtttagc agcccaatgg aagctgagct ggtgcgtcgt 420

attctgtctc gtgacgcagc accgctgcca cgtccgggtg cggttctgaa agattttacc 480

gtgagcggca acctgctgtt tattcgtctg accgcggcag atcatcgtca gctgcaactg 540

agcattagca gctgcctgca acagctgtct ctgctgatgt ggattaccca gtgctttctg 600

ccggtgtttc tggctcaggc gccgtctggt cagcgtcgtc accaccacca ccaccactaa 660

<210>70

<211>219

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>70

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gly

100 105 110

Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro

115 120 125

Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg

130 135 140

Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr

145 150 155 160

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

165 170 175

Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu

180 185 190

Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro

195 200 205

Ser Gly Gln Arg Arg His His His His His His

210 215

<210>71

<211>210

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<220>

<221>PEPTIDE

<222>(0)...(0)

<223>Lage 1b

<400>71

Met Gln Ala Glu Gly Gln Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp

1 5 10 15

Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly

20 25 30

Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala

35 40 45

Gly Ala Ala Arg Ala Ser Gly Pro Arg Gly Gly Ala Pro Arg Gly Pro

50 55 60

His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala

65 70 75 80

Arg Arg Pro Asp Ser Arg Leu Leu Gln Leu His Ile Thr Met Pro Phe

85 90 95

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

100 105 110

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

115 120 125

Ser Gly Asn Leu Leu Phe Met Ser Val Arg Asp Gln Asp Arg Glu Gly

130 135 140

Ala Gly Arg Met Arg Val Val Gly Trp Gly Leu Gly Ser Ala Ser Pro

145 150 155 160

Glu Gly Gln Lys Ala Arg Asp Leu Arg Thr Pro Lys His Lys Val Ser

165 170 175

Glu Gln Arg Pro Gly Thr Pro Gly Pro Pro Pro Pro Glu Gly Ala Gln

180 185 190

Gly Asp Gly Cys Arg Gly Val Ala Phe Asn Val Met Phe Ser Ala Pro

195 200 205

His Ile

210

<210>72

<211>978

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>72

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcggccg cacatatggg tgcgcgtggt 360

ccggaaagcc gtctgctgga attttatctg gccatgccgt ttgcgacccc gatggaagcg 420

gaactggccc gtcgtagcct ggctcaagat gcaccgccgc tgccggttcc gggcgtgctg 480

ctgaaagaat ttaccgtgag cggcaacatt ctgaccattc gtctgacggc ggcagaccat 540

cgtcagctgc aactgagcat tagcagctgc ctgcaacagc tgtctctgct gatgtggatt 600

acccagtgct ttctgccggt gtttctggcc cagccgccgt ctggtcaacg tggtggcgcg 660

cgtcgtccgg attctcgcct gctggaactg catattacca tgccgttcag ctctccaatg 720

gaggccgaat tagtgcgtcg cattctgagc cgtgatgcgg caccgctgcc gcgtccaggt 780

gcggttctga aagacttcac cgtatctggc aacctgctgt ttatccgtct gaccgcagcg 840

gaccaccgcc aattacaatt atctatcagc tcttgtttac aacaactgtc gctgttaatg 900

tggatcactc aatgtttcct gccagtattc ctggctcagg ccccgagcgg tcagcgtcgt 960

caccaccacc accaccac 978

<210>73

<211>326

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>73

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Ala Ala His Met Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe

115 120 125

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

130 135 140

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

145 150 155 160

Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr

165 170 175

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

180 185 190

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

195 200 205

Leu Ala Gln Pro Pro Ser Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp

210 215 220

Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met

225 230 235 240

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

245 250 255

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

260 265 270

Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser

275 280 285

Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln

290 295 300

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

305 310 315 320

His His His His His His

325

<210>74

<211>1209

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>74

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcggccg cacatatgca ggcggaaggc 360

cgtggcaccg gtggtagcac cggcgatgcg gatggtccgg gcggtccggg tattccggac 420

gggcctggtg gtaatgcggg tgggccaggt gaagcgggtg cgaccggtgg tcgtggtccg 480

cggggggcag gcgcagcacg tgcatctggt ccgggtggtg gtgcaccgcg cggtccgcat 540

ggtggtgcgg cgagcggcct gaatggttgc tgccgttgcg gtgcgcgtgg tccggaaagc 600

cgtctgctgg aattttatct ggccatgccg tttgcgaccc cgatggaagc ggaactggcc 660

cgtcgtagcc tggctcaaga tgcaccgccg ctgccggttc cgggcgtgct gctgaaagaa 720

tttaccgtga gcggcaacat tctgaccatt cgtctgacgg cggcagacca tcgtcagctg 780

caactgagca ttagcagctg cctgcaacag ctgtctctgc tgatgtggat tacccagtgc 840

tttctgccgg tgtttctggc ccagccgccg tctggtcaac gtggtggcgc gcgtcgtccg 900

gattctcgcc tgctggaact gcatattacc atgccgttca gctctccaat ggaggccgaa 960

ttagtgcgtc gcattctgag ccgtgatgcg gcaccgctgc cgcgtccagg tgcggttctg 1020

aaagacttca ccgtatctgg caacctgctg tttatccgtc tgaccgcagc ggaccaccgc 1080

caattacaat tatctatcag ctcttgttta caacaactgt cgctgttaat gtggatcact 1140

caatgtttcc tgccagtatt cctggctcag gccccgagcg gtcagcgtcg tcaccaccac 1200

caccaccac 1209

<210>75

<211>403

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>75

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Ala Ala His Met Gln Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly

115 120 125

Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly

130 135 140

Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro

145 150 155 160

Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro

165 170 175

Arg Gly Pro His Gly Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg

180 185 190

Cys Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala

195 200 205

Met Pro Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu

210 215 220

Ala Gln Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu

225 230 235 240

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

245 250 255

His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser

260 265 270

Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln

275 280 285

Pro Pro Ser Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu

290 295 300

Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu

305 310 315 320

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

325 330 335

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

340 345 350

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

355 360 365

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

370 375 380

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

385 390 395 400

His His His

<210>76

<211>981

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>76

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcggccg cacatatggg tgcgcgtcgt 360

ccggatagcc gtctgctgga gctgcatatt accatgccgt ttagcagccc aatggaagct 420

gagctggtgc gtcgtattct gtctcgtgac gcagcaccgc tgccacgtcc gggtgcggtt 480

ctgaaagatt ttaccgtgag cggcaacctg ctgtttattc gtctgaccgc ggcagatcat 540

cgtcagctgc aactgagcat tagcagctgc ctgcaacagc tgtctctgct gatgtggatt 600

acccagtgct ttctgccggt gtttctggct caggcgccgt ctggtcagcg tcgtggtggt 660

gcccgtggcc cggaatctcg tctgctggaa ttttatctgg ccatgccgtt cgcgacgccg 720

atggaagcag agctggcccg tcgcagcctg gctcaggatg caccgccgct gccggttccg 780

ggcgtgctgc tgaaagaatt tacggttagc ggtaacattc tgaccatccg tctgaccgca 840

gcggaccacc gccaactgca actgtctatc agctcttgcc tgcaacaact gtcgttatta 900

atgtggatca ctcaatgttt tttaccagta ttcctggccc aaccgccgag cggccaacgt 960

cgtcaccacc accaccacca c 981

<210>77

<211>327

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>77

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Ala Ala His Met Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu

115 120 125

His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg

130 135 140

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

145 150 155 160

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

165 170 175

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

180 185 190

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

195 200 205

Leu Ala Gln Ala Pro Ser Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro

210 215 220

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

225 230 235 240

Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro

245 250 255

Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn

260 265 270

Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu

275 280 285

Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr

290 295 300

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

305 310 315 320

Arg His His His His His His

325

<210>78

<211>1212

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>78

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa accgcggccg cacatatgca ggcggaaggc 360

cgtggtactg gcggtagcac cggcgatgca gatggtccgg gcggtccggg tattccggat 420

ggtccgggtg gtaatgcagg tggtccaggt gaagcaggtg cgactggcgg tcgtggtcca 480

cgcggtgcag gtgcagcgcg tgcatctggt ccaggtggcg gtgcgccgcg tggcccgcat 540

ggtggtgcag ctagtgcgca agatggtcgt tgcccgtgtg gtgcgcgtcg tccggatagc 600

cgtctgctgg agctgcatat taccatgccg tttagcagcc caatggaagc tgagctggtg 660

cgtcgtattc tgtctcgtga cgcagcaccg ctgccacgtc cgggtgcggt tctgaaagat 720

tttaccgtga gcggcaacct gctgtttatt cgtctgaccg cggcagatca tcgtcagctg 780

caactgagca ttagcagctg cctgcaacag ctgtctctgc tgatgtggat tacccagtgc 840

tttctgccgg tgtttctggc tcaggcgccg tctggtcagc gtcgtggtgg tgcccgtggc 900

ccggaatctc gtctgctgga attttatctg gccatgccgt tcgcgacgcc gatggaagca 960

gagctggccc gtcgcagcct ggctcaggat gcaccgccgc tgccggttcc gggcgtgctg 1020

ctgaaagaat ttacggttag cggtaacatt ctgaccatcc gtctgaccgc agcggaccac 1080

cgccaactgc aactgtctat cagctcttgc ctgcaacaac tgtcgttatt aatgtggatc 1140

actcaatgtt ttttaccagt attcctggcc caaccgccga gcggccaacg tcgtcaccac 1200

caccaccacc ac 1212

<210>79

<211>404

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>79

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Ala

100 105 110

Ala Ala His Met Gln Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly

115 120 125

Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly

130 135 140

Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro

145 150 155 160

Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro

165 170 175

Arg Gly Pro His Gly Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro

180 185 190

Cys Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr

195 200 205

Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu

210 215 220

Ser Arg Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp

225 230 235 240

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

245 250 255

His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser

260 265 270

Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln

275 280 285

Ala Pro Ser Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg

290 295 300

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

305 310 315 320

Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val

325 330 335

Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr

340 345 350

Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser

355 360 365

Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe

370 375 380

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

385 390 395 400

His His His His

<210>80

<211>918

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>80

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgcaggcgga aggccgtggt actggcggta gcaccggcga tgcagatggt 120

ccgggcggtc cgggtattcc ggatggtccg ggtggtaatg caggtggtcc aggtgaagca 180

ggtgcgactg gcggtcgtgg tccacgcggt gcaggtgcag cgcgtgcatc tggtccaggt 240

ggcggtgcgc cgcgtggccc gcatggtggt gcagctagtg cgcaagatgg tcgttgcccg 300

tgtggtgcgc gtcgtccgga tagccgtctg ctggagctgc atattaccat gccgtttagc 360

agcccaatgg aagctgagct ggtgcgtcgt attctgtctc gtgacgcagc accgctgcca 420

cgtccgggtg cggttctgaa agattttacc gtgagcggca acctgctgtt tattcgtctg 480

accgcggcag atcatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 540

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggctcaggc gccgtctggt 600

cagcgtcgtg gtggtgcccg tggcccggaa tctcgtctgc tggaatttta tctggccatg 660

ccgttcgcga cgccgatgga agcagagctg gcccgtcgca gcctggctca ggatgcaccg 720

ccgctgccgg ttccgggcgt gctgctgaaa gaatttacgg ttagcggtaa cattctgacc 780

atccgtctga ccgcagcgga ccaccgccaa ctgcaactgt ctatcagctc ttgcctgcaa 840

caactgtcgt tattaatgtg gatcactcaa tgttttttac cagtattcct ggcccaaccg 900

ccgagcggcc aacgtcgt 918

<210>81

<211>306

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>81

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gln Ala Glu Gly Arg Gly Thr Gly

20 25 30

Gly Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp

35 40 45

Gly Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly

50 55 60

Gly Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly

65 70 75 80

Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Ala Gln Asp

85 90 95

Gly Arg Cys Pro Cys Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu

100 105 110

Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val

115 120 125

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

130 135 140

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

145 150 155 160

Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu

165 170 175

Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val

180 185 190

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

195 200 205

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

210 215 220

Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro

225 230 235 240

Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly

245 250 255

Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln

260 265 270

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

275 280 285

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

290 295 300

Arg Arg

305

<210>82

<211>684

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>82

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgggtgcgcg tggtccggaa agccgtctgc tggaatttta tctggccatg 120

ccgtttgcga ccccgatgga agcggaactg gcccgtcgta gcctggctca agatgcaccg 180

ccgctgccgg ttccgggcgt gctgctgaaa gaatttaccg tgagcggcaa cattctgacc 240

attcgtctga cggcggcaga ccatcgtcag ctgcaactga gcattagcag ctgcctgcaa 300

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggcccagccg 360

ccgtctggtc aacgtggtgg cgcgcgtcgt ccggattctc gcctgctgga actgcatatt 420

accatgccgt tcagctctcc aatggaggcc gaattagtgc gtcgcattct gagccgtgat 480

gcggcaccgc tgccgcgtcc aggtgcggtt ctgaaagact tcaccgtatc tggcaacctg 540

ctgtttatcc gtctgaccgc agcggaccac cgccaattac aattatctat cagctcttgt 600

ttacaacaac tgtcgctgtt aatgtggatc actcaatgtt tcctgccagt attcctggct 660

caggccccga gcggtcagcg tcgt 684

<210>83

<211>228

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>83

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gly Ala Arg Gly Pro Glu Ser Arg

20 25 30

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

35 40 45

Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val

50 55 60

Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr

65 70 75 80

Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser

85 90 95

Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe

100 105 110

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

115 120 125

Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe

130 135 140

Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp

145 150 155 160

Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val

165 170 175

Ser Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln

180 185 190

Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met

195 200 205

Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser

210 215 220

Gly Gln Arg Arg

225

<210>84

<211>915

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>84

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgcaggcgga aggccgtggc accggtggta gcaccggcga tgcggatggt 120

ccgggcggtc cgggtattcc ggacgggcct ggtggtaatg cgggtgggcc aggtgaagcg 180

ggtgcgaccg gtggtcgtgg tccgcggggg gcaggcgcag cacgtgcatc tggtccgggt 240

ggtggtgcac cgcgcggtcc gcatggtggt gcggcgagcg gcctgaatgg ttgctgccgt 300

tgcggtgcgc gtggtccgga aagccgtctg ctggaatttt atctggccat gccgtttgcg 360

accccgatgg aagcggaact ggcccgtcgt agcctggctc aagatgcacc gccgctgccg 420

gttccgggcg tgctgctgaa agaatttacc gtgagcggca acattctgac cattcgtctg 480

acggcggcag accatcgtca gctgcaactg agcattagca gctgcctgca acagctgtct 540

ctgctgatgt ggattaccca gtgctttctg ccggtgtttc tggcccagcc gccgtctggt 600

caacgtggtg gcgcgcgtcg tccggattct cgcctgctgg aactgcatat taccatgccg 660

ttcagctctc caatggaggc cgaattagtg cgtcgcattc tgagccgtga tgcggcaccg 720

ctgccgcgtc caggtgcggt tctgaaagac ttcaccgtat ctggcaacct gctgtttatc 780

cgtctgaccg cagcggacca ccgccaatta caattatcta tcagctcttg tttacaacaa 840

ctgtcgctgt taatgtggat cactcaatgt ttcctgccag tattcctggc tcaggccccg 900

agcggtcagc gtcgt 915

<210>85

<211>305

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>85

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gln Ala Glu Gly Arg Gly Thr Gly

20 25 30

Gly Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp

35 40 45

Gly Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly

50 55 60

Gly Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly

65 70 75 80

Gly Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Gly Leu Asn

85 90 95

Gly Cys Cys Arg Cys Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu

100 105 110

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

115 120 125

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

130 135 140

Leu Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu

145 150 155 160

Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu

165 170 175

Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val

180 185 190

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

195 200 205

Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro

210 215 220

Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala Ala Pro

225 230 235 240

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

245 250 255

Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu

260 265 270

Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr

275 280 285

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

290 295 300

Arg

305

<210>86

<211>687

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>86

atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac 60

gacaagcata tgggtgcgcg tcgtccggat agccgtctgc tggagctgca tattaccatg 120

ccgtttagca gcccaatgga agctgagctg gtgcgtcgta ttctgtctcg tgacgcagca 180

ccgctgccac gtccgggtgc ggttctgaaa gattttaccg tgagcggcaa cctgctgttt 240

attcgtctga ccgcggcaga tcatcgtcag ctgcaactga gcattagcag ctgcctgcaa 300

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggctcaggcg 360

ccgtctggtc agcgtcgtgg tggtgcccgt ggcccggaat ctcgtctgct ggaattttat 420

ctggccatgc cgttcgcgac gccgatggaa gcagagctgg cccgtcgcag cctggctcag 480

gatgcaccgc cgctgccggt tccgggcgtg ctgctgaaag aatttacggt tagcggtaac 540

attctgacca tccgtctgac cgcagcggac caccgccaac tgcaactgtc tatcagctct 600

tgcctgcaac aactgtcgtt attaatgtgg atcactcaat gttttttacc agtattcctg 660

gcccaaccgc cgagcggccaacgtcgt 687

<210>87

<211>229

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>87

Met Gly His His His His His His His His His His Ser Ser Gly His

1 5 10 15

Ile Asp Asp Asp Asp Lys His Met Gly Ala Arg Arg Pro Asp Ser Arg

20 25 30

Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala

35 40 45

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

50 55 60

Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser Gly Asn Leu Leu Phe

65 70 75 80

Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser

85 90 95

Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe

100 105 110

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

115 120 125

Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr Leu Ala Met Pro

130 135 140

Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln

145 150 155 160

Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr

165 170 175

Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg

180 185 190

Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu

195 200 205

Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Pro Pro

210 215 220

Ser Gly Gln Arg Arg

225

<210>88

<211>1197

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>88

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa acccaggcgg aaggccgtgg tactggcggt 360

agcaccggcg atgcagatgg tccgggcggt ccgggtattc cggatggtcc gggtggtaat 420

gcaggtggtc caggtgaagc aggtgcgact ggcggtcgtg gtccacgcgg tgcaggtgca 480

gcgcgtgcat ctggtccagg tggcggtgcg ccgcgtggcc cgcatggtgg tgcagctagt 540

gcgcaagatg gtcgttgccc gtgtggtgcg cgtcgtccgg atagccgtct gctggagctg 600

catattacca tgccgtttag cagcccaatg gaagctgagc tggtgcgtcg tattctgtct 660

cgtgacgcag caccgctgcc acgtccgggt gcggttctga aagattttac cgtgagcggc 720

aacctgctgt ttattcgtct gaccgcggca gatcatcgtc agctgcaact gagcattagc 780

agctgcctgc aacagctgtc tctgctgatg tggattaccc agtgctttct gccggtgttt 840

ctggctcagg cgccgtctgg tcagcgtcgt ggtggtgccc gtggcccgga atctcgtctg 900

ctggaatttt atctggccat gccgttcgcg acgccgatgg aagcagagct ggcccgtcgc 960

agcctggctc aggatgcacc gccgctgccg gttccgggcg tgctgctgaa agaatttacg 1020

gttagcggta acattctgac catccgtctg accgcagcgg accaccgcca actgcaactg 1080

tctatcagct cttgcctgca acaactgtcg ttattaatgt ggatcactca atgtttttta 1140

ccagtattcc tggcccaacc gccgagcggc caacgtcgtc accaccacca ccaccac 1197

<210>89

<211>399

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>89

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gln

100 105 110

Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp Gly Pro

115 120 125

Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly Gly Pro

130 135 140

Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala Gly Ala

145 150 155 160

Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro His Gly

165 170 175

Gly Ala Ala Ser Ala Gln Asp Gly Arg Cys Pro Cys Gly Ala Arg Arg

180 185 190

Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser

195 200 205

Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala Ala

210 215 220

Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser Gly

225 230 235 240

Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln

245 250 255

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

260 265 270

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

275 280 285

Arg Arg Gly Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe Tyr

290 295 300

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

305 310 315 320

Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val Pro Gly Val Leu Leu

325 330 335

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

340 345 350

Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln

355 360 365

Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu

370 375 380

Ala Gln Pro Pro Ser Gly Gln Arg Arg His His His His His His

385 390 395

<210>90

<211>1194

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>90

atggatccaa gcagccattc atcaaatatg gcgaataccc aaatgaaatc agacaaaatc 60

attattgctc accgtggtgc tagcggttat ttaccagagc atacgttaga atctaaagca 120

cttgcgtttg cacaacaggc tgattattta gagcaagatt tagcaatgac taaggatggt 180

cgtttagtgg ttattcacga tcacttttta gatggcttga ctgatgttgc gaaaaaattc 240

ccacatcgtc atcgtaaaga tggccgttac tatgtcatcg actttacctt aaaagaaatt 300

caaagtttag aaatgacaga aaactttgaa acccaggcgg aaggccgtgg caccggtggt 360

agcaccggcg atgcggatgg tccgggcggt ccgggtattc cggacgggcc tggtggtaat 420

gcgggtgggc caggtgaagc gggtgcgacc ggtggtcgtg gtccgcgggg ggcaggcgca 480

gcacgtgcat ctggtccggg tggtggtgca ccgcgcggtc cgcatggtgg tgcggcgagc 540

ggcctgaatg gttgctgccg ttgcggtgcg cgtggtccgg aaagccgtct gctggaattt 600

tatctggcca tgccgtttgc gaccccgatg gaagcggaac tggcccgtcg tagcctggct 660

caagatgcac cgccgctgcc ggttccgggc gtgctgctga aagaatttac cgtgagcggc 720

aacattctga ccattcgtct gacggcggca gaccatcgtc agctgcaact gagcattagc 780

agctgcctgc aacagctgtc tctgctgatg tggattaccc agtgctttct gccggtgttt 840

ctggcccagc cgccgtctgg tcaacgtggt ggcgcgcgtc gtccggattc tcgcctgctg 900

gaactgcata ttaccatgcc gttcagctct ccaatggagg ccgaattagt gcgtcgcatt 960

ctgagccgtg atgcggcacc gctgccgcgt ccaggtgcgg ttctgaaaga cttcaccgta 1020

tctggcaacc tgctgtttat ccgtctgacc gcagcggacc accgccaatt acaattatct 1080

atcagctctt gtttacaaca actgtcgctg ttaatgtgga tcactcaatg tttcctgcca 1140

gtattcctgg ctcaggcccc gagcggtcag cgtcgtcacc accaccacca ccac 1194

<210>91

<211>398

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>91

Met Asp Pro Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

1 5 10 15

Ser Asp Lys Ile Ile Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

20 25 30

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

35 40 45

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

50 55 60

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

65 70 75 80

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

85 90 95

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Gln

100 105 110

Ala Glu Gly Arg Gly Thr Gly Gly Ser Thr Gly Asp Ala Asp Gly Pro

115 120 125

Gly Gly Pro Gly Ile Pro Asp Gly Pro Gly Gly Asn Ala Gly Gly Pro

130 135 140

Gly Glu Ala Gly Ala Thr Gly Gly Arg Gly Pro Arg Gly Ala Gly Ala

145 150 155 160

Ala Arg Ala Ser Gly Pro Gly Gly Gly Ala Pro Arg Gly Pro His Gly

165 170 175

Gly Ala Ala Ser Gly Leu Asn Gly Cys Cys Arg Cys Gly Ala Arg Gly

180 185 190

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

195 200 205

Pro Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro

210 215 220

Pro Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly

225 230 235 240

Asn Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln

245 250 255

Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile

260 265 270

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

275 280 285

Arg Gly Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile

290 295 300

Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile

305 310 315 320

Leu Ser Arg Asp Ala Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys

325 330 335

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

340 345 350

Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu

355 360 365

Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala

370 375 380

Gln Ala Pro Ser Gly Gln Arg Arg His His His His His His

385 390 395

<210>92

<211>867

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>92

atgcatcatc atcatcatca ccaggcggaa ggccgtggta ctggcggtag caccggcgat 60

gcagatggtc cgggcggtcc gggtattccg gatggtccgg gtggtaatgc aggtggtcca 120

ggtgaagcag gtgcgactgg cggtcgtggt ccacgcggtg caggtgcagc gcgtgcatct 180

ggtccaggtg gcggtgcgcc gcgtggcccg catggtggtg cagctagtgc gcaagatggt 240

cgttgcccgt gtggtgcgcg tcgtccggat agccgtctgc tggagctgca tattaccatg 300

ccgtttagca gcccaatgga agctgagctg gtgcgtcgta ttctgtctcg tgacgcagca 360

ccgctgccac gtccgggtgc ggttctgaaa gattttaccg tgagcggcaa cctgctgttt 420

attcgtctga ccgcggcaga tcatcgtcag ctgcaactga gcattagcag ctgcctgcaa 480

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggctcaggcg 540

ccgtctggtc agcgtcgtgg tggtgcccgt ggcccggaat ctcgtctgct ggaattttat 600

ctggccatgc cgttcgcgac gccgatggaa gcagagctgg cccgtcgcag cctggctcag 660

gatgcaccgc cgctgccggt tccgggcgtg ctgctgaaag aatttacggt tagcggtaac 720

attctgacca tccgtctgac cgcagcggac caccgccaac tgcaactgtc tatcagctct 780

tgcctgcaac aactgtcgtt attaatgtgg atcactcaat gttttttacc agtattcctg 840

gcccaaccgc cgagcggcca acgtcgt 867

<210>93

<211>289

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>93

Met His His His His His His Gln Ala Glu Gly Arg Gly Thr Gly Gly

1 5 10 15

Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly

20 25 30

Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly

35 40 45

Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly

50 55 60

Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Ala Gln Asp Gly

65 70 75 80

Arg Cys Pro Cys Gly Ala Arg Arg Pro Asp Ser Arg Leu Leu Glu Leu

85 90 95

His Ile Thr Met Pro Phe Ser Ser Pro Met Glu Ala Glu Leu Val Arg

100 105 110

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

115 120 125

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

130 135 140

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

145 150 155 160

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

165 170 175

Leu Ala Gln Ala Pro Ser Gly Gln Arg Arg Gly Gly Ala Arg Gly Pro

180 185 190

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

195 200 205

Met Glu Ala Glu Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro

210 215 220

Leu Pro Val Pro Gly Val Leu Leu Lys Glu Phe Thr Val Ser Gly Asn

225 230 235 240

Ile Leu Thr Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu

245 250 255

Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr

260 265 270

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

275 280 285

Arg

<210>94

<211>633

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>94

atgcatcatc atcatcatca cggtgcgcgt ggtccggaaa gccgtctgct ggaattttat 60

ctggccatgc cgtttgcgac cccgatggaa gcggaactgg cccgtcgtag cctggctcaa 120

gatgcaccgc cgctgccggt tccgggcgtg ctgctgaaag aatttaccgt gagcggcaac 180

attctgacca ttcgtctgac ggcggcagac catcgtcagc tgcaactgag cattagcagc 240

tgcctgcaac agctgtctct gctgatgtgg attacccagt gctttctgcc ggtgtttctg 300

gcccagccgc cgtctggtca acgtggtggc gcgcgtcgtc cggattctcg cctgctggaa 360

ctgcatatta ccatgccgtt cagctctcca atggaggccg aattagtgcg tcgcattctg 420

agccgtgatg cggcaccgct gccgcgtcca ggtgcggttc tgaaagactt caccgtatct 480

ggcaacctgc tgtttatccg tctgaccgca gcggaccacc gccaattaca attatctatc 540

agctcttgtt tacaacaact gtcgctgtta atgtggatca ctcaatgttt cctgccagta 600

ttcctggctc aggccccgag cggtcagcgt cgt 633

<210>95

<211>211

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>95

Met His His His His His His Gly Ala Arg Gly Pro Glu Ser Arg Leu

1 5 10 15

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

20 25 30

Leu Ala Arg Arg Ser Leu Ala Gln Asp Ala Pro Pro Leu Pro Val Pro

35 40 45

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

50 55 60

Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser

65 70 75 80

Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu

85 90 95

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

100 105 110

Arg Pro Asp Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser

115 120 125

Ser Pro Met Glu Ala Glu Leu Val Arg Arg Ile Leu Ser Arg Asp Ala

130 135 140

Ala Pro Leu Pro Arg Pro Gly Ala Val Leu Lys Asp Phe Thr Val Ser

145 150 155 160

Gly Asn Leu Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu

165 170 175

Gln Leu Ser Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp

180 185 190

Ile Thr Gln Cys Phe Leu Pro Val Phe Leu Ala Gln Ala Pro Ser Gly

195 200 205

Gln Arg Arg

210

<210>96

<211>864

<212>DNA

＜213〉homo sapiens (Homo Sapien)

<400>96

atgcatcatc atcatcatca ccaggcggaa ggccgtggca ccggtggtag caccggcgat 60

gcggatggtc cgggcggtcc gggtattccg gacgggcctg gtggtaatgc gggtgggcca 120

ggtgaagcgg gtgcgaccgg tggtcgtggt ccgcgggggg caggcgcagc acgtgcatct 180

ggtccgggtg gtggtgcacc gcgcggtccg catggtggtg cggcgagcgg cctgaatggt 240

tgctgccgtt gcggtgcgcg tggtccggaa agccgtctgc tggaatttta tctggccatg 300

ccgtttgcga ccccgatgga agcggaactg gcccgtcgta gcctggctca agatgcaccg 360

ccgctgccgg ttccgggcgt gctgctgaaa gaatttaccg tgagcggcaa cattctgacc 420

attcgtctga cggcggcaga ccatcgtcag ctgcaactga gcattagcag ctgcctgcaa 480

cagctgtctc tgctgatgtg gattacccag tgctttctgc cggtgtttct ggcccagccg 540

ccgtctggtc aacgtggtgg cgcgcgtcgt ccggattctc gcctgctgga actgcatatt 600

accatgccgt tcagctctcc aatggaggcc gaattagtgc gtcgcattct gagccgtgat 660

gcggcaccgc tgccgcgtcc aggtgcggtt ctgaaagact tcaccgtatc tggcaacctg 720

ctgtttatcc gtctgaccgc agcggaccac cgccaattac aattatctat cagctcttgt 780

ttacaacaac tgtcgctgtt aatgtggatc actcaatgtt tcctgccagt attcctggct 840

caggccccga gcggtcagcg tcgt 864

<210>97

<211>288

<212>PRT

＜213〉homo sapiens (Homo Sapien)

<400>97

Met His His His His His His Gln Ala Glu Gly Arg Gly Thr Gly Gly

1 5 10 15

Ser Thr Gly Asp Ala Asp Gly Pro Gly Gly Pro Gly Ile Pro Asp Gly

20 25 30

Pro Gly Gly Asn Ala Gly Gly Pro Gly Glu Ala Gly Ala Thr Gly Gly

35 40 45

Arg Gly Pro Arg Gly Ala Gly Ala Ala Arg Ala Ser Gly Pro Gly Gly

50 55 60

Gly Ala Pro Arg Gly Pro His Gly Gly Ala Ala Ser Gly Leu Asn Gly

65 70 75 80

Cys Cys Arg Cys Gly Ala Arg Gly Pro Glu Ser Arg Leu Leu Glu Phe

85 90 95

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

100 105 110

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

115 120 125

Leu Lys Glu Phe Thr Val Ser Gly Asn Ile Leu Thr Ile Arg Leu Thr

130 135 140

Ala Ala Asp His Arg Gln Leu Gln Leu Ser Ile Ser Ser Cys Leu Gln

145 150 155 160

Gln Leu Ser Leu Leu Met Trp Ile Thr Gln Cys Phe Leu Pro Val Phe

165 170 175

Leu Ala Gln Pro Pro Ser Gly Gln Arg Gly Gly Ala Arg Arg Pro Asp

180 185 190

Ser Arg Leu Leu Glu Leu His Ile Thr Met Pro Phe Ser Ser Pro Met

195 200 205

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

210 215 220

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

225 230 235 240

Leu Phe Ile Arg Leu Thr Ala Ala Asp His Arg Gln Leu Gln Leu Ser

245 250 255

Ile Ser Ser Cys Leu Gln Gln Leu Ser Leu Leu Met Trp Ile Thr Gln

260 265 270

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

275 280 285

<210>98

<211>450

<212>PRT

＜213〉artificial sequence

＜223〉3-protein d-MAGE-A3-His

<400>98

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

1 5 10 15

Ala Gly Cys Ser Ser His Ser Ser Asn Met Ala Asn Thr Gln Met Lys

20 25 30

Ser Asp Lys Ile I1e Ile Ala His Arg Gly Ala Ser Gly Tyr Leu Pro

35 40 45

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

50 55 60

Tyr Leu Glu Gln Asp Leu Ala Met Thr Lys Asp Gly Arg Leu Val Val

65 70 75 80

Ile His Asp His Phe Leu Asp Gly Leu Thr Asp Val Ala Lys Lys Phe

85 90 95

Pro His Arg His Arg Lys Asp Gly Arg Tyr Tyr Val Ile Asp Phe Thr

100 105 110

Leu Lys Glu Ile Gln Ser Leu Glu Met Thr Glu Asn Phe Glu Thr Met

115 120 125

Asp Leu Glu Gln Arg Ser Gln His Cys Lys Pro Glu Glu Gly Leu Glu

130 135 140

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

145 150 155 160

Glu Glu Gln Glu Ala Ala Ser Ser Ser Ser Thr Leu Val Glu Val Thr

165 170 175

Leu Gly Glu Val Pro Ala Ala Glu Ser Pro Asp Pro Pro Gln Ser Pro

180 185 190

Gln Gly Ala Ser Ser Leu Pro Thr Thr Met Asn Tyr Pro Leu Trp Ser

195 200 205

Gln Ser Tyr Glu Asp Ser Ser Asn Gln Glu Glu Glu Gly Pro Ser Thr

210 215 220

Phe Pro Asp Leu Glu Ser Glu Phe Gln Ala Ala Leu Ser Arg Lys Val

225 230 235 240

Ala Glu Leu Val His Phe Leu Leu Leu Lys Tyr Arg Ala Arg Glu Pro

245 250 255

Val Thr Lys Ala Glu Met Leu Gly Ser Val Val Gly Asn Trp Gln Tyr

260 265 270

Phe Phe Pro Val Ile Phe Ser Lys Ala Ser Ser Ser Leu Gln Leu Val

275 280 285

Phe Gly Ile Glu Leu Met Glu Val Asp Pro Ile Gly His Leu Tyr Ile

290 295 300

Phe Ala Thr Cys Leu Gly Leu Ser Tyr Asp Gly Leu Leu Gly Asp Asn

305 310 315 320

Gln Ile Met Pro Lys Ala Gly Leu Leu Ile Ile Val Leu Ala Ile Ile

325 330 335

Ala Arg Glu Gly Asp Cys Ala Pro Glu Glu Lys Ile Trp Glu Glu Leu

340 345 350

Ser Val Leu Glu Val Phe Glu Gly Arg Glu Asp Ser Ile Leu Gly Asp

355 360 365

Pro Lys Lys Leu Leu Thr Gln His Phe Val Gln Glu Asn Tyr Leu Glu

370 375 380

Tyr Arg Gln Val Pro Gly Ser Asp Pro Ala Cys Tyr Glu Phe Leu Trp

385 390 395 400

Gly Pro Arg Ala Leu Val Glu Thr Ser Tyr Val Lys Val Leu His His

405 410 415

Met Val Lys Ile Ser Gly Gly Pro His Ile Ser Tyr Pro Pro Leu His

420 425 430

Glu Trp Val Leu Arg Glu Gly Glu Glu Gly Gly His His His His His

435 440 445

His His

450

Claims (29)

1. fusion rotein, it comprises:

(a) NY-ESO-1 or its fragment, it is connected to

(b) LAGE-1 or its fragment,

Wherein at least a among NY-ESO-1 and/or the LAGE-1 be brachymemma or the part brachymemma, or comprise the fragment of one or more epi-positions of NY-ESO-1 or LAGE-1.

2. according to the fusion rotein of claim 1, wherein said NY-ESO-1 is selected from: the NY-ESO-1 of total length NY-ESO-1, part brachymemma or the NY-ESO-1 of brachymemma or its any fragment, described fragment comprises one or more epi-positions of NY-ESO-1.

3. according to the fusion rotein of any aforementioned claim, wherein said LAGE-1 is selected from: the LAGE-1 of total length LAGE-1, part brachymemma or the LAGE-1 of brachymemma or its any fragment, described fragment comprises one or more epi-positions of LAGE-1.

4. according to the fusion rotein of any aforementioned claim, wherein said NY-ESO-1 or LAGE-1 and naturally occurring NY-ESO-1 or LAGE-1 at least 95,96,97,98,99% or 100% are equal to.

5. according to the fusion rotein of any aforementioned claim, wherein said LAGE-1 is LAGE-1a.

6. according to the fusion rotein of any aforementioned claim, the N-terminal of wherein said NY-ESO-1 and the C-terminal of described LAGE-1 merge.

7. according to each fusion rotein among the claim 1-5, the C-terminal of wherein said NY-ESO-1 and the N-terminal of described LAGE-1 merge.

8. according to the fusion rotein of any aforementioned claim, wherein said fusion rotein further comprises the allos fusion partner.

9. fusion rotein according to Claim 8, wherein said allos fusion partner is 3-protein d or derivatives thereof or fragment.

10. according to the fusion rotein of claim 9, the pact that wherein said 3-protein d derivative comprises 3-protein d is preceding 1/3, for example the amino acid 20-127 of 3-protein d.

11. according to the fusion rotein of claim 9 or 10, wherein said 3-protein d derivative is not a lipidization.

12. according to the fusion rotein of any aforementioned claim, it further comprises amino acid Met, Asp and Pro.

13. according to the fusion rotein of claim 12, wherein said amino acid Met, Asp and Pro and merge according to the N-terminal of each 3-protein d allos fusion partner among the claim 9-11.

14. according to the fusion rotein of any aforementioned claim, wherein said fusion rotein is a recombination fusion protein.

15. according to the fusion rotein of any aforementioned claim, wherein said fusion rotein further comprises affinity tag.

16. according to the fusion rotein of claim 15, wherein said affinity tag is the Histidine tail that comprises 1-10 histidine residues.

17. according to each fusion rotein among the claim 1-5, it comprises and is selected from following aminoacid sequence: SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ IDNO:8, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:15, SEQ IDNO:16, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:23, SEQID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ IDNO:46, SEQ ID NO:48, SEQ ID NO:51, SEQ ID NO:53, SEQID NO:55, SEQ ID NO:57, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81, SEQ ID NO:83, SEQ ID NO:85, SEQ IDNO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:93, SEQID NO:95 and SEQ ID NO:97.

18. according to each fusion rotein among the claim 1-5, it comprises and is selected from following aminoacid sequence: SEQ ID NO:81, SEQ ID NO:85, SEQ ID NO:93 and SEQID NO:97.

19. a nucleic acid molecule, each fusion rotein among its coding claim 1-18.

20. a carrier, it comprises the nucleic acid molecule of claim 19.

21. a host cell, its carrier with claim 20 transforms.

22. immunogenic composition or vaccine, its comprise according among the claim 1-18 each fusion rotein, as the nucleic acid molecule that requires in the claim 19 or as the carrier of requirement in the claim 20.

23. as the immunogenic composition that requires in the claim 22, it comprises adjuvant and/or the immunostimulatory cell factor or chemokine in addition.

24. as the immunogenic composition or the vaccine that require in claim 22 or 23, wherein said fusion rotein is present in oil-in-water or the water-in-oil emulsion vehicle.

25. as the immunogenic composition or the vaccine that require in claim 23 or 24, it comprises one or more following adjuvant: 3D-MPL, QS21 or CpG oligonucleotide.

26. as the immunogenic composition or the vaccine that require in each among the claim 22-26, it comprises one or more other antigens in addition.

27. as the immunogenic composition or the vaccine that require in each among the claim 22-26, it is used for using in medical science.

28. the composition of the carrier of the fusion rotein of claim 1-18 or the nucleic acid molecule of claim 19 or claim 20 or claim 22-26 or vaccine are used for the treatment of purposes in the medicine of cancer in preparation, described cancer is the mammary gland melanoma for example; Mammary cancer; Prostate cancer; Bladder cancer comprises transitional cell carcinoma; Lung cancer comprises nonsmall-cell lung cancer (NSCLC); The H﹠N cancer comprises the esophageal carcinoma; Squamous cell carcinoma; Gastrointestinal cancer; Liver cancer; Cerebral tumor; Leukemia; With various sarcomas.

29. the purposes of claim 28 is used for the treatment of for the underproof patient of Her2/neu targeted therapy.

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