USE OF HAPTEN-CARRIER COMPLEXES FOR BENZIMIDAZOLE PESTICIDES IMMUNOASSAYS DEVELOPMENT

摘要

In order to develop immunoassays for pesticides detection, this work describes the choice of three different haptensthat present structural similarity to benzimidazole molecule, methods for coupling them with two carrier proteins inorder to make them immunogenic, a protocol for immunization of laboratory rodents with hapten-carrier complexes,and the evaluation of the specific antibody responses against haptens using an in-house developed immunoassay. Threecarbendazim (methyl 2-benzimidazole-carbamate) derivatives bearing different functional reactive groups (-NH2, -SHand -COOH), namely 2-(2-Aminoethyl) benzimidazole (AEB),2-Mercaptobenzimidazole (2MB) and 2-Benzimidazolepropionic acid (BPA), were coupled to keyhole limpet haemocyanin (KLH) and bovine serum albumin(BSA),respectively, mixed with immuno-adjuvants, and injected four times into Balb/C mice and Wistar rats for induction ofspecific immune responses.All three chemicals elicited a specific but weak antibodies response upon immunization withhapten-KLH complexes followed by serological testing by indirect ELISA with hapten-BSA complexes, and showeddetectabledifferences in antibody titers with regard to number of inoculations, hapten structure and animal species.Whereas the AEB-KLH complex was the strongest, the 2MB-KLH complex was the weakest immunogenic in mice.However, the best animal responders allow the application of technologies for getting monoclonal antibodies againstbenzimidazoles, which can then be used for immunoassays development.