An enzyme-linked immunosorbent assay for aconitine-type alkaloids using an anti-aconitine monoclonal antibody
An enzyme-linked immunosorbent assay for aconitine-type alkaloids using an anti-aconitine monoclonal antibody
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DOI：
10.1016/j.aca.2008.04.002
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发表时间：
2008-05-26
影响因子：
6.2
通讯作者：
Shoyama, Yukihiro
中科院分区：
文献类型：
--
作者：
Kido, Katsumi;Edakuni, Kyoko;Shoyama, Yukihiro
将3-琥珀酰乌头碱与牛血清白蛋白(BSA)偶联，制备抗乌头碱(ACO)的单抗。将ACO-BSA结合物免疫的小鼠脾细胞与氨基喋呤敏感的小鼠骨髓瘤细胞系P3-X63-Ag8-653融合，成功地获得了分泌抗ACO单抗的杂交瘤细胞株。单抗与ACO类生物碱中乌头碱、次乌头碱和新乌头碱发生交叉反应，但不与其他化合物发生交叉反应。使用新的单抗开发的酶联免疫吸附试验(ELISA)的全部测量范围从100ngmL(-1)扩大到1.5mgmLACO。用该方法测定了不同附子中ACO类生物碱的含量，结果与文献报道基本一致。(C)2008爱思唯尔B.V.保留所有权利。
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 ngmL(-1) to 1.5 mu g 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. (C) 2008 Elsevier B.V. All rights reserved.