脱氧雪腐镰刀菌烯醇人工抗原和多克隆抗体的制备

Preparation of artificial antigen and polyclonal antibodies against deoxynivalenol

对脱氧雪腐镰刀菌烯醇(DON)的两种半抗原合成方法进行对比研究,合成半抗原,并采用活化酯法将半抗原与牛血清白蛋白偶联制备得到免疫原,通过特定程序免疫日本大耳白兔获得了多克隆抗体。抗血清效价为1∶32000,protei-nA-sepharose4B亲和层析纯化的DON抗体与DON、3-AC-DON(3-乙酰氧基脱氧雪腐镰刀菌烯醇)、15-AC-DON(15-乙酰氧基脱氧雪腐镰刀菌烯醇)、T-2毒素、OTA(赭曲霉A)、ZEN(玉米赤霉烯酮)的交叉反应率分别为:100%、500%、2.50%、0.10%、0.01%、0.01%,表明了所制备DON抗体的高特异性。

Deoxnivalenol hapten was synthesized successfully followed by studying of the two kinds of hapten synthesis method. The immunogen was prepared following a scheme that employs active ester method for the conjugation of the haptens to BSA（bovine serum albumin）, with which polyclonal antibodies against deoxynivalenol were raised in rabbits after immunization. The antiserum titer was 1∶32 000, and the antiserum was purified by proteinA-sepharose 4B immunity affinity chromatography. The antibody specificity was checked on the basis of the 50% inhibition levels, relative cross-reactivities with DON, 3-AC-DON, 15-AC-DON, T-2, OTA and ZEN were found to be 100%, 500%, 2.50%, 0.10%, 0.01%, 0.01%.