恩诺沙星完全抗原的合成及兔源多抗血清的制备  

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

作　　者：刘莉 张家禾[2] 袁维峰[3] 孟婷 

机构地区：[1]江苏省兽用生物制药高技术研究重点实验室,江苏泰州225300 [2]江西农业大学动物科学技术学院,江西南昌330045 [3]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《中国畜牧兽医》2014年第6期38-41,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家高技术研究发展计划(863计划)(2012AA101302);安徽省科技攻关项目(1201c0602006);江苏省"青蓝工程"中青年学术带头人项目;公益性行业(农业)科研专项(200903027;201303042);中央级公益性科研院所基本科研业务费专项资金项目(2011js-3)

摘　　要：为了合成恩诺沙星（enrofloxacin，EFLX）完全抗原，制备兔源多克隆抗体血清，试验通过活化酯法将小分子的EFLX偶联到牛血清白蛋白（BSA）和鸡卵清白蛋白（OVA）上，形成偶联物EFLX-BSA和EFLX—OVA，以EFLX—BSA为免疫原免疫兔，以EFLXOVA为检测原通过ELISA方法检测兔血清效价。结果表明，免疫兔血清效价都在1：51200以上，其中2号兔效价最高，达到1：204800，敏感性好，半数抑制浓度IC50为169．91ng／mL，检测范围为37．52～302．31ng／mL。本试验成功制备了EFLX完全抗原，获得了敏感的兔源多克隆抗体血清，为以后ELISA试剂盒和胶体金试纸条的研发奠定了基础。The purpose of this paper was to synthesize complete antigen of enrofloxacin （EFLX）, and to prepare the rabbit potyclonalantibody. Ovalbumin （OVA） and bovine serum albumin （BSA） were used as protein carriers to synthesize small mole- cule 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 ser- um. This experiment obtained high antibody titer,which could lay the foundation for the development of the ELISA kit and colloidal gold test strip.

关 键 词：恩诺沙星 残留 检测 活化酯法 间接竞争ELISA 

分 类 号：S859.84[农业科学—临床兽医学]