马波沙星人工抗原的合成及其免疫原性的鉴定  

Synthesizing artificial antigen of marbofloxacin and identifying immunogenicity of the antigent

作　　者：汝晓飞 张伟[1] 王兴[1] 秦海艳[1] 周小钰 侯珂[1] 刘若璇[1] 蔡云虹[1] 王捍东[1] RU Xiaofei;ZHANG Wei;WANG Xing;QIN Haiyan;ZHOU Xiaoyu;HOU Ke;LIU Ruoxuan;CAI Yunhong;WANG Handong(College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Chin)

机构地区：[1]扬州大学兽医学院

出　　处：《畜牧与兽医》2018年第4期66-71,共6页Animal Husbandry & Veterinary Medicine

基　　金：江苏高校优势学科建设工程资助项目(PAPD);江苏省动物重要疫病与人兽共患病防控协同创新中心;江苏省高校重点实验室开放课题资助;江苏省普通高校自然科学基金(08KJD230002)

摘　　要：合成、鉴定了马波沙星（MBF）人工抗原,为马波沙星单克隆抗体的制备及其免疫学检测方法的建立奠定基础。以牛血清白蛋白（BSA）、鸡卵清蛋白（OVA）为载体蛋白,采用碳二亚胺法分别合成马波沙星免疫抗原（MBF-BSA）和包被抗原（MBF-OVA）,并使用紫外分光光度法、SDS-PAGE、ELISA方法鉴定人工抗原合成效果。初步鉴定结果显示,MBF-BSA与MBF-OVA均偶联成功;用免疫抗原MBF-BSA免疫小鼠,经ELISA方法检测,五免后血清中抗体效价可达1∶5.12×105,半数抑制率为90.20 ng/m L。研究表明,马波沙星人工抗原合成成功,其免疫抗原具有良好的免疫原性,免疫小鼠可获得高效价、高特异性多克隆抗体。The present study was to synthesize and identify the artificial antigen of marbofloxacin and to lay foundation for preparation of monoclonal antibodies of marbofloxacin and establishment of immunological detection methods. MBA-BSA and MBF-OVA were synthesized using the carbodiimide method,with bovine serum albumin（BSA） and chicken ovalbumin（OVA） as carrier proteins. The synthetic effect of artificial antigens was observed using UV spectrophotometry,SDS-PAGE,ELISA. The results of preliminary examination showed that MBFBSA and MBF-OVA were coupled successfully by UV spectrophotometry and SDS-PAGE. Immunizing mice with MBF-BSA produced specificity antibodies against the MBF in serum and the antibody titer was detected to be 1： 5. 12 × 10~5 by ELISA. In conclusion,the successfully synthesized antigens of marbofloxacin had good immunogenicity and could produce high titer and high specificity polyclonal antibody by immunizing mice.

关 键 词：马波沙星 人工抗原 免疫原性 兽药残留 

分 类 号：S859.84[农业科学—临床兽医学] S852.43[农业科学—兽医学]