Production of monoclonal antibodies and enzyme immunoassay for typical adenylate cyclase activator,Forskolin |
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| Authors: | Ritsu Sakata Yukihiro Shoyama Hiroki Murakami |
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| Affiliation: | (1) Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashiku, 812 Fukuoka, Japan;(2) Graduate School of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, 812 Fukuoka, Japan |
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| Abstract: | 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  g), GLC (30 ng) and HPLC (1 g) methods. |
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| Keywords: | Adenylate cyclase activator Coleus forskohlii enzyme immunoassay forskolin monoclonal antibody mass spectrometry |
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