Preparation Of The Anti-Diethylatibestrol Monoclonal Antibody And Enzyme-Labelled Antigen

Diethylstibestrol (DES) is an artificial estrogen. It have a good effect in improving the metabolism of protein, the growth of the animal and decreasing the fat, so was used in the feeding of the animal.The International Agency for Research on Cancer(IARC) have found: DES can lead to cancer, and can residues in animal products, such as liver, meats, egg and milk, and hazard health through food chain. So it was ruled not to use DES in feeding animal by any way all over the world. Animals and animal products could not be detected DES.The residual immunoassay of DES is an important measure to monitor the use of DES. The aim of this research is to develop the DES residual immunoassay regent box. It included the contents: The preparation and characterization of DES artificial antigens; the preparation of the polyclonal and monoclonal antibody of DES; The synthesis of HRP-DES and the method of the enzyme-linked immunosorbent assay (ELISA) were established to detect DES residual.Diethylstibestol-4-carboxymethyl ether was synthesized through the method of organic chemistry. And it was successfully synthesized by the results of the MS, IR, and H-NMR. Mixed anhydride method and Isobutylchlorofomate and carbodiimide method were used to synthesis DES artifical antigen, which respectively conjugated DES-4-carboxymethyl ether to Bovine serum albumin(BSA) and Ovabumin(OVA), four DES artificial antigens were right materials by identification of ultraviolet spectrophotometry(UV) and sodium doclecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE). The number of hapten molecules per molecules of protein estimated was 30 and 18 respectively.BSA-DES synthesized by mixed anhydride method was used as immunogen to immunized rabbits, and OVA-DES synthesized respectively mixed anhydride method and Isobutylchlorformate and carbodiimide method were immunized mice, and three anti-sera...