Objective:The goal of this study was to synthesize artificial antigen of AFB1 and obtain its mice polyclonal antiserum. Method:Immunogen AFB1-BSA and coating antigen AFB1-OVA were synthesized using NHS by linking carrier proteins BSA and OVA to AFB1 and identified by ultraviolet scanning,SDS-PAGE. The mice polyclonal antiserum was obtained by BALB/C mice immunization and was identified by indirect ELISA and blocking ELISA. Result:The results showed that the hapten AFB1 was successfully linked to carrier proteins. Indirect ELISA showed a high titre above 1×10-4 of the antiserum of all the six BALB/c mice. The sensitivity of antiserum of No. 3 mice was the best,which IC50 was 8.199 ng/mL. Conclusion:Artificial antigen AFB1-BSA and AFB1-OVA were synthesized and sensitivity AFB1 polyclonal antiserum has been generated in this study,which laid a foundation base for the preparation of monoclonal antibodies and rapid test reagent against AFB1.