Synthesis of carbendazim artificial antigens and preparation of murine polyclonal antibodies
Objective To synthesize carbendazim artificial antigens,prepare carbendazim polyclonal antibodies and identify their characteristics.Methods Active carboxyl groups were introduced to prepare the carbendazim haptens by the mixed anhydride method.The artificial antigens and coating antigens were obtained by coupling the small molecule haptens with carriers of bovine serum albumin(BSA)and ovalbumin(OVA).Sodium dodecyl sulfate polycrylamide gel electropheresis(SDS-PAGE)was used to identify carbendazim artificial antigens.Mice were immunized with the prepared artificial antigens to obtain polyclonal antibodies against carbendazim,and the antibody titers and specificity were identified by indirect ELISA.Results Carbendazim artificial antigens were successfully prepared.The titer of polyclonal antibody was above 1∶12 800 and the half-maximal inhibitory concentration(IC50)of the antibody was 0.107 μg/mL.The cross-reactivity rates with both benomyl and thiabendazole were less than 1％.Conclusion Polyclonal antibodies with high sensitivity and high specificity were successfully prepared,laying the foundation for the establishment of a rapid detection method for carbendazim residues.
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