2000 Fiscal Year Final Research Report Summary
MEDICAL APPLICATION AND DEVELOPMENT OF EPITOPE DETECTION SYSTEM TO ANTIGEN RELATED AUTO IMMUNE AND ALLERGY DISORDERS
| Project/Area Number |
09557218
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Laboratory medicine
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| Research Institution | NAGOYA CITY UNIVERSITY |
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Principal Investigator |
KUNIMATSU Mitoshi NAGOYA CITY UNIVERSITY, MEDICAL SCHOOL, LECTURER, 医学部, 講師 (70145746)
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| Co-Investigator(Kenkyū-buntansha) |
OHTSUKA Takanobu NAGOYA CITY UNIVERSITY, MEDICAL SCHOOL, Assistant Professor, 医学部, 講師 (10185316)
TADA Toyohiro NAGOYA CITY UNIVERSITY, NURSING SCHOOL, PROFESSOR, 看護学部, 教授 (20106230)
ITO Hirotaka NAGOYA CITY UNIVERSITY, MEDICAL SCHOOL, ASSOSIATE PROFESSOR, 医学部, 助教授 (60080115)
OZAKI Yasuhiko NAGOYA CITY UNIVERSITY, MEDICAL SCHOOL, RESEACH ASSOSIATE, 医学部, 助手 (50254280)
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| Project Period (FY) |
1997 – 2000
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| Keywords | epitope / auto antigen / auto antibody / peptide chip / macro array / immuno-ome |
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| Research Abstract |
A concept of this research is to develop the peptide chips including amino acid sequences of a target proteins to be related to a disease. We attempted medical application and development of epitope detection system to antigen related auto immune and allergy disorders by using this eptide chips. B cell epitopes to calpain, calpastatin and fodrin were analyzed with peptide chips on ce llulose membranes spotted a small amount of peptides. The antibody to each epitope obviously stained the neuronal ischemia regions and myocardial infraction regions. Epitopes to autoantigen CENP-A and Epiplakin were analyzed with its autoimmune disease patient serum. Specific amino acid sequences on these auto antigens were found by using autoimmune disease patient sera. This assay system was useful for screening and diagnosis of autoimmune disease. We succeeded in a producing the peptide chips more over 2,000 knds of peptides on membrane approximately five times, compared with conventional methods. Macro array machine could detect super very small amount of antigens with a fluorescent antibody as probe. This system applied to an antigen dot assay methods. The protein antigen spotted on nitrocellulose membrane, the antigen chip manufactured. Each spot was a very small amount of antigen, 60-to 160 nL/spot. We tried to detect an anti TSH auto antibody in autoimmune disease patient serum. A strong immune reaction obtained from TSH alpha subunit but not its beta subunit. This system was available for kinds of 160,000-tetra peptide library including D-amino acids and amino acid derivatives.
It was able to apply a development of various ferment prevention agent. Now we attempt to make high-density peptide chips synthesis system at high speed further.
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