Detection of antigen epitope for bullous pemphigois antigenic protein and the development of autoantibody detection using synthetic polypeptide
| Project/Area Number |
02557045
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Dermatology
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| Research Institution | Keio University |
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Principal Investigator |
NISHIKAWA Takeji Keio University School of Medicine, Professor, 医学部, 教授 (50051579)
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| Co-Investigator(Kenkyū-buntansha) |
HASEGAWA Akira Tonen Corporate R & D Laboratory, Senior Investigator, 研究員
MIKI Keizaburo Tonen Corporate R & D Laboratory, Director, 所長
TANAKA Masaru Keio Univ. Sch. Med., Instructor, 医学部, 助手 (40188339)
HASHIMOTO Takashi Keio university School of Medicine, Assist. Prof., 医学部, 講師 (20129597)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥10,600,000 (Direct Cost: ¥10,600,000)
Fiscal Year 1991: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1990: ¥9,100,000 (Direct Cost: ¥9,100,000)
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| Keywords | bullous pemphigoid / anti-basement membrane zone antibodies / immunoblotting / antigenic epitopes for epidermal basement zone / 自己免疫性水疱瘡 / 抗表性基底膜部抗体 / リコンビナント(基底膜抗原)蛋白 / 自己免疫性疾患 / 自己抗体 / エピト-プ / ELISA / 分子生物学 |
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| Research Abstract |
The final destination of this developmental scientific research was to detect anti-basement membrane zone (BMZ) antibodies from patients with bullous pemphigoid (BP) using a newly synthesized polypeptide antigen that contains epitope for BMZ antigen protein.
To approach this purpose, we have collected as many sera as possible from Japan, U. K. and U. S. patients with BP. We have found that the BMZ antibodies from different continents are similarly heterogenous in terms of antigenic epitope as assessed by immunofluorescence, immunoblotting and immunoprecipitation. Then, we have characterized BP sera by use of recombinant BP antigen proteins and tested the possibility of applying ELISA technique. At the same time we specified the epitope of the"230 kD BP antigen for human anti-BMZ monoclonal antibody 5E by means of deletion mutants of the mouse CDNA Clone that encodes for about carboxyl-terminal half of t. he 230 kD BP antigen protein. The epitope for 5E was found to localize in the 114 amino acid residues encoded by Bgl 11 - StuI fragment of the CDNA. From these studies the following conclusions were obtained ; (1) there are several epitopes present in the BEIZ antigens and (2) more than 3 synthetic peptides are necessary to cover the entire epitopes for ELISA technique to detect 230 kD antigen only. More investigations are needed to reach the final destination.
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Report
(3 results)
Research Products
(16 results)
