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An enzyme-linked immunosorbent assay for aconitine-type alkaloids using an anti-aconitine monoclonal antibody
Authors:Kido Katsumi  Edakuni Kyoko  Morinaga Osamu  Tanaka Hiroyuki  Shoyama Yukihiro
Institution:

aGraduate School of Pharmaceutical Sciences, Kyushu University, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan

bFaculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7, Huis Ten Bosch, Sasebo City, Nagasaki 859-3298, Japan

Abstract:3-Succinylaconitine was conjugated with bovine serum albumin (BSA) for use as an immunogen for the preparation of a monoclonal antibody (MAb) against aconitine (Aco). Splenocytes from mice immunized with the Aco-BSA conjugate were fused with an aminopterin-sensitive mouse myeloma cell line, P3-X63-Ag8-653, and a hybridoma secreting a MAb against Aco was successfully obtained. The MAb cross-reacted with mesaconitine, hypaconitine and jesaconitine, which are Aco-type alkaloids, but not with any other compounds examined. The full measurement range of an enzyme-linked immunosorbent assay (ELISA) developed using the new MAb extended from 100 ng mL−1 to 1.5 μg mL−1 of Aco. The concentrations of Aco-type alkaloids in various Aconiti radixes assayed using the new ELISA method showed good agreement with previous reports.
Keywords:Aconitine-type alkaloid  Monoclonal antibody  Enzyme-linked immunosorbent assay  Aconiti radix
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