PRODUCTION OF MONOCLONAL-ANTIBODIES AND ENZYME-IMMUNOASSAY FOR TYPICAL ADENYLATE-CYCLASE ACTIVATOR, FORSKOLIN

被引:66
作者
SAKATA, R
SHOYAMA, Y
MURAKAMI, H
机构
[1] KYUSHU UNIV, FAC PHARMACEUT SCI, DEPT PHARMACOGNOSY, HIGASHI KU, FUKUOKA 812, JAPAN
[2] KYUSHU UNIV, FAC AGR, GRAD SCH GENET RESOURCES TECHNOL, HIGASHI KU, FUKUOKA 812, JAPAN
关键词
ADENYLATE CYCLASE ACTIVATOR; COLEUS FORSKOHLII; ENZYME IMMUNOASSAY; FORSKOLIN; MONOCLONAL ANTIBODY; MASS SPECTOMETRY;
D O I
10.1007/BF00754612
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The ratio of hapten to bovine serum albumin in an antigen conjugate was exactly determined by matrix-assisted laser desorption/ionization mass spectrometry. A hybridoma secreting monoclonal antibodies against forskolin was produced by fusing splenocytes immunized with a forskolin-bovine serum albumin conjugate with HAT-sensitive mouse myeloma cells. The cross-reaction of anti-forskolin antibodies with 7-deacetyl forskolin was 5.57%. A very small cross-reaction appeared with 1-deoxy, 9-deoxy and 1,9-dideoxy forskolin derivatives. The full measuring range of the assay extends from 6 ng to 200 ng of forskolin. The competitive ELISA assay used for this analysis was found to be more sensitive than TLC (10 mu g), GLC (30 ng) and HPLC (1 mu g) methods.
引用
收藏
页码:101 / 108
页数:8
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