| Project/Area Number |
06557016
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Human pathology
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| Research Institution | Fukui Medical School |
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Principal Investigator |
FUKUDA Masaru Faculty of medicine, Fukui Medical School Professor, 医学部, 教授 (60079720)
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| Co-Investigator(Kenkyū-buntansha) |
SASAMOTO Masami DOJINDO LAB., Research Division.Chairman, 研究部, 研究部長
KOHNO Masahiro JEOL.Co.Ltd., Analysis Division.Chairman, 分析機器本部, グループ長
NORIKI Sakon Medical School Hospital, Fukui Medicial School Research assistant, 医学部附属病院, 助手 (30228374)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1994: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | BIOTIN / TEMPO / SPIN-ASSAY / SPIN-IMMUNOASSAY / ESR / AVIDIN / BIOTIN-TEMPO / BIOCYTIN-TEMPO / ビチオン・TEMPO / ビオチン・TEMPO |
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| Research Abstract |
We synthesized biotin TEMPO.The ESR signals of biotin-TEMPO consist of three sharp lines of nitroxide radical of TEMPO.The signals of biotin TEMPO was decreased in proportion to the amount of Avidin. We proved that avidin has no dismutase activity of TEMPO,and that decrease of biotin-TEMPO signals don't result from increasing protein concentration. Attenuating of the signals of biotin-TEMPO in proportion to the amount of Avidin is result from the combination avidin and biotin-TEMPO.In order to dissociate avidin-biotin complex, we treated with autoclave, 120 ﾟC,40 minutes. The TEMPO signals were recovered by this heat treatment. The peak height of recovered signals was about 50% of that before conjugation with avidin. Autoclave treatment didn't attenuate signals of 4-amino-TEMPO.Moreover, the recovered signals of TEMPO by autoclave treatment was again decreased by the addition of avidin. These facts show that avidin-biotin-TEMPO complex is dissociated by autoclave treatment, and biotin-TEMPO is not destructed by autoclave treatment. Autoclave don't degrade biotin-TEMPO to biotin and TEMPO.We can try to test varied dissociation conditions of avidin biotin complex. This may make clear the reason of very strong combination of avidin and biotin. We examined basic research of avidin quantification ; the detection limits of avidin activity is 0.012mg/ml and CV value is 4.1%, co-relation value (gamma) is 0.995. Then we measured serum avidin activity of healthy persons (n=4) and patients with inflammatory disease (n=16). The avidin activity of healthy persons is 0.0375 <plus-minus> 0.0038mg/ml.that of patients with inflammation is 0.0413 <plus-minus> 0.0079mg/ml. No significant differences are found. As for as the immno-spinassay, the sensitivity of this methods is too poor to apply immuno-spinassay. During this research, We also synthesized and purified biocytin-TEMPO to determine biotinidase activ
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