Preparation of Conjugate Antigen and Antibody to Chlorpromazine in the Animal Food
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Abstract:
In this study, an enzyme-linked immunosorbent assay (ELISA) for the determination of chlorpromazine residues in animal foods was established. Chlorpromazine hapten (CPZ-H) was synthesized from chlorpromazine by demethylation, reaction with tert-butyl acrylate and hydrolysis for the first time. Hapten CPZ-H was conjugated with OVA and BSA by active ester method to form the coated antigen and immunogen, respectively. New Zealand white rabbits were immunized with immunogen CPZ-H-BSA to successfully prepare specific polyclonal antibodies against chlorpromazine. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for the rapid determination of chlorpromazine was established based on the antibody. By optimizing the working conditions of ic-ELISA method, it was determined that the dilution fold of coated antigen and antibody was 1:8000. At this time, the IC50 of standard curve was 1.1 ng/mL and the linear range was 0.13 ng/mL~8.8 ng/mL. The prepared polyclonal antibody against chlorpromazine has good specificity and no cross reaction with other chlorpromazine analogues. This study provides a basis for the preparation of specific and rapid detection kit for chlorpromazine in the future.
