喹乙醇代谢物完全抗原的合成与多克隆抗体的制备
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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.
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.
引文
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[2] CHEN Q,TANG S,JIN X,ZOU J,et al.Investigation of the genotoxicity of guinocetone,carbadox and olaquindox in vitro using vero cells[J].Food Chem Toxicol,2009,47(2):328.
[3] ZON J J,CHEN Q,JIN X,et al.Olaquindox induces apoptosis through the mitochondrial pathway in Hep G2 cells[J].Toxicology,2011,285(3):104.
[4]中国兽药典委员会.中华人民共和国兽药典兽药使用指南2010年版化学药品卷[S].2010.
[5]王仁华,练小华,谢益根.饲料中喹乙醇的应用效果研究[J].饲料与畜牧:新饲料,2017(2):56.