| Project/Area Number |
06557120
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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 |
Physical pharmacy
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
GOTO Junichi TOHOKU UNIVERSITY FACULTY OF PHARMACEUTICAL SCIENCES PROFESSOR, 薬学部, 教授 (80006337)
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| Co-Investigator(Kenkyū-buntansha) |
NIWA Toshifumi TOHOKU UNIVERSITY COLLEGE OF MEDICAL SCIENCES ASSOCIATE PROFESSOR, 医療技術短大, 助教授 (90218248)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥8,700,000 (Direct Cost: ¥8,700,000)
Fiscal Year 1995: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1994: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | Enzyme immunoassay / Electrospray ionization / LC / MS / 11-Deoxycortisol / Capillary electrophoresis / Monoclonal antibody / リゾチーム / エレエレクトロスプレーイオン化 |
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| Research Abstract |
Recently, many interests have been focused on the usefulness of mass spectrometry (MS) heyphenated with chromatographic methods such as liquid chromatography (LC) and capillary electrophoresis for the structural elucidation of proteins. On the other hand, it is keen needed elucidation of mechanisms of the intermolecular interaction between proteins and biologically active molecules. The purpose of this project was the development of the method for the structural elucidation of the domain of the anti-haptenic antibody bounded to the hapten.
Initially, the monoclonal antibody for 11-deoxycortisol was prepared using the steroid having a bridge at C-4 on the nucleus as a hapten. Next, lysosome as a model enzyme for enzyme immunoassay was reacted with 11-deoxycortisol and the structure of the conjugated lysozyme was analyzed with electrospray ionization (ESI) mass spectrometry on the flow injection mede. Before and after reductive alkylation, the conjugated lysosome has been further subjected to hydrolysis with various enzyme followed by the LC/ESI-MS analysis to clarify the structure of the domain.
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