磺胺嘧啶多克隆抗体的制备

Preparation of the Polyclonal Antibodies against Sulfadiazine

为制备磺胺嘧啶(sulfadiazine,SD)多克隆抗体(pAb),用重氮化方法将SD偶联于载体蛋白BSA和OVA上,合成免疫抗原BSA-SD和包被抗原OVA-SD。用紫外扫描、SDS-PAGE凝胶电泳试验检测,结果表明偶联成功。用BSA-SD免疫新西兰大白兔,间接ELISA测定多克隆抗体效价。阻断ELISA结果显示:SD单克隆抗体(pAb)的IC50达19.19ng/mL,和其他磺胺类药物交叉反应小于等于2.7%。结果表明,获得了高效价和特异的pAb。

To prepare the polyclonal antibodies aginst sulfadiazine, the artificial immunizing antigen BSA - SD and coating antigen OVA - SD obtained by diazotization, and further identified by ultraviolet （UV） and SDS- PAGE. New Zealand white rabbits were immunized with BSA - SD. The titres of polyclonal antibodies were detected by indirect ELISA and blocking ELISA. The results showed that the IC50 reached to 19.19 ng/mL and the cross reaction with other sulfa drugs was less than or equal to 2.7%. This indicated that the SD polyclonal antibodies of high titer,sensitivity and specificity has been generated and made it possible to establish immunoassay of SD residues.