Abstract:

The complete antigen of AFB1 was synthesized. The mouse polyclonal antiserum was prepared by immunizing the BALB/c mice with the complete antigen. The carboxyl group was introduced in the Aflatoxin B1 (AFB1) molecule by Succinimide ester method. AFB1O was produced. AFB1-BSA and AFB1-OVA were synthesized by EDC and DCC methods. Thin layer chromatography technology, Uv spectroscopy, SDS-PAGE and immunological methods were used to identify the complete antigen. BALB/c mice were immunized with AFB1-BSA. The titer, sensibility and specificity of the antiserum was determined by indirect ELISA and competitive ELISA. The results showed that the antiserum titer of all the BALB/c mice were over 1:3200. The highest No.6 mouse reached 1.28×10-4 and the sensitivity is good with half inhibitory concentration IC50 of 22.268 ng/mL, low cross-reactivity. The complete antigen of AFB1was synthesized successfully. The sensitive polyclonal antiserum of AFB1 has been gained in this study, which laid a foundation for the preparation of monoclonal antibodies and the establishment of a rapid detection method against AFB1.