| Project/Area Number |
60580138
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Osaka City University |
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Principal Investigator |
MINAMIURA Noshi Faculty of Science, Osaka City University, Professor, 理学部, 教授 (20047129)
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| Co-Investigator(Kenkyū-buntansha) |
IIZUKA Masaru Faculty of Science, Osaka City University Lecturere, 理学部, 講師 (00047292)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1987: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1986: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1985: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Human Pancreatic <alpha>-Amylase / Human Salivary (alpha)-Amylase / Pancreatic <alpha>-Amylase Specific Monoclonal Antibody / モノクローナル抗体 / 分別定量 |
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| Research Abstract |
This Study had been done in order to make it clear the structures of antigenic binding sits of human salivary and pacreatic <alph>-amylase specific monoclonal antibodies and also to develop the immunoassay system using them for diagnosis.
1. Human salivary <alph>-amylase specific monoclonal antibody--- Only two mouse hybridoma strains producing salivary <alph>-amylase specific monoclonal antibody (IgG) were isolated by the cell fusion repeated more than 10 times, and then a sandwich enzyme immunoassay method was made up to determine discriminatory salivary -amyulase in serum with a concetration of 0:1 to 10 umole per ml of the enzyme protein. The assay system was found to be not suitable for diagosis because of low sensitivity. The another cell fusion experiments is continuously done to get some hybridomas which produce more active nomoclonal antibodies.
2. Human pancreatic <alph>-amylase specific monoclonal antibody--- Several mouse hybridomas producing specific monoclonal antibody were obtained by the fusion repeated more than several decade times. Using the monoclonal antibody produced by them, an effective enzyme immunoassay system was setablished to determine discriminatory pancreatic <alph>-amylase with high sensitivity of assaying a concentration of 60 f mole per ml of the enzyme. At the present time, the system is examined for the development in some hospitals.
3. Structural study of the specific monoclonal antibodies--- It took long time to get hybridoma strains which produced the specific monoclonal antibody against salivary as well as pancreatic <alph>-amylases because the appearence of the hybridomas was very low, as descrived above. This study are still in progress.
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