乌头碱免疫抗原及多克隆抗体的制备

Preparation of Immunogen and Polyclonal Antibodies against Aconitine

乌头碱与戊二酸酐反应得半抗原乌头碱-3-基戊二酸单酯,并采用活泼酯法使其与牛血清白蛋白(BSA)结合,制得乌头碱免疫抗原(AC-GH-BSA),经MALDI-TOF质谱分析其蛋白偶联率为18。以此抗原免疫新西兰兔,制得的多克隆抗体能够特异性结合乌头碱,并采用间接酶联免疫吸附法测定抗体效价。结果显示,制备的乌头碱多克隆抗体效价为1∶1 280 000。

The hapten, mono aconitine-3-yl glutarate （AC-GH）, was synthesized by introduction of a glutaryl group to aconitine （AC）. The derivative was then conjugated with bovine serum albumin （BSA） by the active ester method to form the artificial antigen （AC-GH-BSA）. Eighteen molecules of AC-GH were coupled with one molecule of BSA determined by MALDI-TOF MS. Polyclonal antibody （PAbs） was produced from immunized New Zealand rabbits and could bind to AC specifically. The titer of PAbs was 1 ： 1 280 000.