对位红抗原合成和多克隆抗体的制备

Synthesis of an Antigen of Para Red and Production of Polyclonal Antibody Specific for Para Red

为制备一种特异性强的抗对位红的多克隆抗体,采用混合酸酐法将活化的对位红半抗原与阳离子化牛血清白蛋白(cBSA)偶联成免疫原,免疫大白兔制备多克隆抗体,利用紫外分光光度计分析结合物的结合情况,间接ELISA和间接竞争ELISA分析抗体特性。发现紫外扫描分析的免疫原对位红与蛋白结合的偶联率为14:1,间接竞争ELISA法分析多克隆抗体效价达到1×106,50%抑制率检测限为7.43ng/mL,检测的对位红线性范围在0.1ng/mL^1μg/mL,抗体与对位红和苏丹红Ⅰ交叉反应率分别为100%和97.8%,与苏丹红Ⅱ、Ⅲ、Ⅳ和甲苯胺红的交叉反应率很低,与罗丹明类色素无交叉反应。说明制备的对位红多克隆抗体具有很高的特异性,与其他色素交叉反应率低,可用于对位红在食品残留中的检测。

To prepare a polyclonal antibody （pAb） specific for para red, hapten para red was coupled with carrier protein of cationed bovine serum albumin （cBSA） to form a complete antigen by mixed anhydride methods. The conjugates used to immunize Japanese white rabbits were identified with UV scanning spectrum and the specificity of pAb was detected by indirect ELISA and in indirect compete ELISA（icELISA）. The couple rate of the antigen was 14：1, the titer of pAb was 1×10 6 and the detection limit in term of 50% inhibition rate was 7.43 ng/mL in the linear range of 0.1 ng/mL-1 μg/mL. The cross-reaction rate between para red and pAb was 100% and was 97.8% between Sudan I and pAb. There was very lower cross-reaction between pAb and para red analogs. The anti-para red polyclonal antibody had a high specificity and was good enough for immunoassay of para red in edible food products.