喹乙醇代谢物完全抗原的合成与多克隆抗体的制备  

Synthesis and Identification of Olaquindox Metobolite Complete Antigen and Preparation of its Polyclonal Antibody

作　　者：高子函 黄金林 GAO Zihan;HUANG Jinlin(Jiangsu Key Lab of Zoonosis/Jiangsu Co-Innovation Center for Prevention and Control of Important Animal InfectiousDiseases and Zoonoses,Yangzhou University, Yangzhou Jiangsu 225009, China)

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《解放军预防医学杂志》2019年第1期4-7,共4页Journal of Preventive Medicine of Chinese People's Liberation Army

摘　　要：目的制备喹乙醇代谢物3-甲基-喹啉-2-羟酸的多克隆抗体,为食品安全快速检测喹乙醇代谢物提供支持。方法用乙酰甲喹制备喹乙醇代谢物3-甲基-喹啉-2-羧酸半抗原,再经以活化酯法制备喹乙醇代谢物-OVA(OLAOVA)与喹乙醇代谢物-BSA(OLA-BSA),用质谱分析鉴定完全抗原;采用长程免疫法免疫小鼠,用间接ELISA法检测抗体血清效价,用竞争ELISA法检测所得抗体特异性与灵敏度。结果成功制备了喹乙醇代谢物-OVA与喹乙醇代谢物-BSA完全抗原,长效免疫小鼠并获得血清抗体效价均达到1∶104以上。结论经过长程免疫法可以得到高效价抗体血清,为食品安全快速检测提供技术支撑。Objective To synthesize a complete antigen of 3-methyl-quinoline-hydroxy acid(MQHA),to prepare polyclonal antibodies against this olaquindox metobolite,and to provide some experimental data for the immunoassay of MQHA.Methods MQHA was prepared using mequindox.The complete antigen(MQHA-BSA and MQHA-OVA)was prepared by coupled MQHA to BSA and OVA using activated ester method,and identified by MALDI-TOF-MS.BALB/c mice were immunized with MQHA-BSA to produce the antiserum that was detected by indirect ELISA and competitive ELISA for titers,specificity and sensitivity.Results MQHA-OVA and MQHA-BSA were successfully prepared and the coupling ratio was 7:1 and 13:1 for MQHA-OVA and MQHA-BSA,respectively.The animals immunized with MQHA-BSA produced specific antibodies which reacted specifically to MQHA with the titer up to 1:10000.Conclusion A complete antigen has been successfully synthesized and specific antiserumprepared.

关 键 词：喹乙醇代谢物 3-甲基-喹啉-2-羟酸 完全抗原 多克隆抗体 

分 类 号：R185.3[医药卫生—流行病学]