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.