抗腹泻性贝毒软海绵酸多克隆抗体的制备与检测

Development and characterization of polyclonal antibodies against diarrhetic shellfish toxin,okadaic acid

制备了腹泻性贝毒软海绵酸(okadaic acid,0A)的多克隆抗体,鉴定了抗体特性。利用半抗原BSA偶联小分子毒素OA,制备完全抗原OA-BSA,免疫两只新西兰白兔,分离、纯化抗血清。用间接ELISA方法测定抗体效价,两个兔抗血清效价均达到128,000以上。IC50为2.852ng/mL。DOT BLOT测定抗体特异性,结果表明,抗体与抗原有特异性反应。试验表明,抗OA多克隆抗体制备成功,其质量较好,可用于将来的免疫检测研究。

To prepare and analyze the polyclonal antibodies against diarrhetic shellfish toxin, （DSP）, okadaic acid （OA） ,complete antigen OA-BSA was made by conjugating the small molecules toxin OA to carrier BSA. Two rabbits were immunized with the OA-BSA conjugate and the antiserum was purified and by saturated sul furic ammonium precipitation method, followed by affinity chromatography. The affinity constant was determined, and the IC50 value was measured by ELISA. The sensitivity was tested by DOT BLOT. The results showed that the titers of the antiserum is over 128 000 and IC50 value is about from 0. 11 ng/mL to 1ng/mL. The result of DOT BLOT indicates specific reaction between the antigen and antibodies. This test shows polyclonal antibodies may be useful for the future research.