Preparation Of Artificial Antigen Of Diethylstilbestrol And Establishment And Application Of ELISA For Detection

Diethylstilbestrol (DES), which was prepared artificially, had been used in the animal husbandry extensively as the growing promoter. And the studies in broad and at home showed that DES had severe side effects, could destroy the inherent materials and caused the gene mutation to form tumor. At present, the use of DES had been forbidden in the animal husbandry and aquatic culture. But the abuse of DES is still present. For the ensuring of human safety and health, it is necessary to establish a rapid, sensitive method to detect the residue of DES in the animal derived food.Using the DES, succinic anhydride and bovine serum albumin (BSA) as the main materials, DES-succinate was obtained by the acylation of succinic anhydride. Then the DES-succinate and BSA were connected through mixed acid anhydride reaction (isobutyl chloroformate method) to obtain the immunogen. The immuogen was connected with ovalbumin to obtain coating antigen. The rabbits were vaccinated by the immunogen, and the antibody titer was detected by the indirect ELISA. The results showed that the antibody titer could reach 28 , which demonstrated the good immunogenity of artificial antigen and could stimulate the rabbits to secrete the expected antibody of high level.The indirect ELISA to detect antibody against DES, which was sensitive and specific, was established. The optimum conditions were established through the matrix test. The optimum concentration of coating antigen was 9.6μg/ml; the best coating effect could be reached at the condition of 4℃,24h,and the best blocking effect could be reached at the condition of...