Production of monoclonal antibodies and enzyme immunoassay for typical adenylate cyclase activator, Forskolin.
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 micrograms), GLC (30 ng) and HPLC (1 microgram) methods.
Sakata
R
R
Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Shoyama
Y
Y
Murakami
H
H
D000262:Adenylyl Cyclases / Q000187:drug effects*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D005576:Colforsin / Q000032:analysis*; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D007124:Immunoenzyme Techniques; D007834:Lasers; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D012710:Serum Albumin, Bovine / Q000737:chemistry
D000911:Antibodies, Monoclonal; D005576:Colforsin; D012710:Serum Albumin, Bovine; D000262:Adenylyl Cyclases