| Project/Area Number |
62870011
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
General pharmacology
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| Research Institution | Kitasato University School of Medicine |
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Principal Investigator |
KATORI Makoto Kitasato University School of Medicine Prof., 医学部, 教授 (50050365)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Shozo Tokushima University School of Medicine Prof., 医学部, 教授 (50025607)
MUROTA Sei-itsu Tokyo Medical and Dental University School of Dentistry Prof., 歯学部, 教授 (50072989)
TADA Michihiko Osaka University School of Medicine Prof., 医学部, 教授 (90093434)
SATOH Kazuo Nihon University School of Medicine Prof., 医学部, 教授 (80010180)
ABE Keishi Tohoku University School of Medicine Prof., 医学部, 教授 (60004777)
鶴藤 〓 東北大学, 薬学部, 教授 (40012596)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥12,100,000 (Direct Cost: ¥12,100,000)
Fiscal Year 1989: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1988: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1987: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | Thromboxane A_2 / 11-dehydro-thromboxane B_2 / Immunoaffinity-columu / 11ーDHーTXB_2 / 抗体固定化カラム / 尿 / 血漿 / 動物種差 / オ-プン体 / 11ーデヒドローTXB_2 / 抗体固相化カラム / 2,3ージノールーTXB_2 |
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| Research Abstract |
TXB2 had been used as an index of TXA2 generation in the body, but cannot be used anymore, because of its artifactual generation during blood collection. Instead, 11- dehydro(DH)-TXB2 was chosen and its enzyme immunoassay method was developed by a research member. The subsequent usage of the commercially available RIA kits prompted us to proceed analyses of the determination method of 11-DH-TXB2 and we realized that the extraction and purification was an unavoidable step before assay. Then, the simpler extraction and purification procedure by an immunoaffinity column was explored icolaboration with Sankyo group of this research group. The monoclonal antibody against 11- DH-TXB2, produced by Dr. Yamamoto's group, Tokushima University, was used for this immunoaffinity column and the standard extraction method was established by a workshop of this research group. Each member of this research group measured the 11-DH-TXB2 content in the distributed known samples and the column was applied
… More
to measure ll-DH- TXB2 levels in pathological models and in clinical patients. It was confirmed that this immunoaffinity columns was useful for assay of 11-DH-TXB2 not only in urine, but also in plasma. There are, however, many difficulties for taking 11-DH-TXB2 as an index of TXA2; the large variety in animal species differences, showing that dogs scarcely produce ll-DH- TXB2 from TXB2. In contrast, 11-DH-TXB2 is produced largely in human from TXB2 and its half life is approximately 60 min, indicating that this metabolite can be taken as an index of TXA2 generation in clinical field. Moreover, we found many difficulties in this assay, such as use of the open type of 11-DH-TXB2, method to exclude substances, disturbed in RIA procedures, reliability of isotope labeled standards, available commercially (large variation of the contentents in individual lots) and others. It is clear that three years studies of this research group established the assay method of 11-DH-TXB2 including extraction and purification using this immunoaffinity column and disclosed many difficulties during the extraction. Less
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