酸性红73多克隆抗体的制备及其应用

在酸性红73分子的羟基上引入一个带有羧基的“间隔臂”,采用N-羟基琥珀亚胺活性酯法将酸性红73分别与牛血清白蛋白(BSA)、卵清蛋白(OA)偶联,合成免疫原和包被原,经免疫新西兰白兔获得多克隆抗体,所得抗体最大效价可达2.56×105,建立了酸性红73的间接竞争ELISA检测方法.本方法的半数抑制浓度(IC50)为181.2 μg/L,检出限(LOD)为7.9 μg/L.交叉反应实验表明,除苏丹红3号(1.13％)外,抗AR73抗体与其它竞争物均无交叉反应.在虾仁中的空白添加回收率为63.5％～90.7％,RSD＜6.8％.说明本方法可用于虾仁中酸性红73的残留检测.

Preparation and Application of Acid Red 73 Polyclonal Antibody

Connecting the hydroxyl of acid red 73(AR73) molecule with a "spacer arm" containing a carboxyl as hapten,and the hapten was conjugated to bovine serum albumin(BSA) or ovalbumin(OA) by using N-hydroxysuccinimide active ester method as immunogen and coating antigen respectively,the polyclonal antibody titer was 2.56×105 through immunization of New zealand white rabbits,then an indirect ELISA procedure for the determination of acid red 73 was established.The half of inhibition concentration(IC50) was 181.2 μg/L,the limit of detection for acid red 73 was 7.9 μg/L,and the cross-reactivity study showed that the polyclonal antiserum was highly specific to AR73,and no cross-reactivity was detected between the obtained polyclonal antiserum and the other competitors except to Sudan red Ⅲ(1.13%).The recoveries from the standards fortified blank samples were in the range of 63.5%-90.7% with RSD lower than 6.8%.The developed method can be used to determine the acid red 73 residue in shrimps,and can help to prepare the monoclonal antibody and develop the rapid test kits for acid red 73.