喹乙醇代谢物完全抗原的合成与多克隆抗体的制备

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。

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

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.