The Ciprofloxacin (CIP) was conjugated directly to bovine serum albumin(BSA) by carbodiimide (EDC) method and isobutyl chloroformate method, giving the immunizing antigen. And the coating antigen, CIP-OVA, was synthesized by EDC method. The conjugated CIP-BSA and CIP-OVA were analyzed by non-denaturing gel electrophoresis and UV spectrophotometry. Results showed that the molecule conjugate ratio of CIP to BSA by EDC method and isobutyl chloroformate method were 4:1 and 11:1, respectively. The molecule conjugate ratio of CIP to OVA was 12:1. The successful synthesis of Ciprofloxacin artificial antigens provided references for establishment of immunoassay of Ciprofloxacin.