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Synthesis of Complete Antigen of Enrofloxacin and Preparation of Rabbit Polyclonal Antiserum

LIU Li1，ZHANG Jia-he2，YUAN Wei-feng3，MENG Ting1   

  1. （1.Jiangsu Key Laboratory for Veterinary Biotechnologic Drug，Taizhou 225300，China；2.College of Animal Science and Technology，Jiangxi Agricultural University，Nanchang 330045，China；3.Institute of Animal Sciences，Chinese Academy of Agricultural Sciences，Beijing 100193，China）

  • Received:2013-11-12 Online:2014-06-20 Published:2014-07-24

Abstract: The purpose of this paper was to synthesize complete antigen of enrofloxacin （EFLX），and to prepare the rabbit polyclonalantibody.Ovalbumin （OVA） and bovine serum albumin （BSA） were used as protein carriers to synthesize small molecule drugs EFLX complete antigens by active ester method.The conjugates formed in this way was EFLX-BSA and EFLX-OVA，the serum titer was detected by use of EFLX-BSA immunogen and EFLX-OVA coating antigen by ELISA.The results showed that the serum titer detection result of three rabbits were all above 1∶51200，the second were the highest of all，its result was 1∶204800 and the sensitive was good.The result of indirect competitive ELISA indicated that this method had a linear detection range from 37.52 to 302.31 ng/mL （R2＝0.9838），with IC50 values of 169.91 ng/mL.In this paper，the immunogen and coating antigen were successfully synthesized，and prepared the good sensetive of rabbit poloclonal antibody from the serum. This experiment obtained high antibody titer，which could lay the foundation for the development of the ELISA kit and colloidal gold test strip.

Key words:

enrofloxacin; residue; detection; active ester method; indirect competitive ELISA