摘要
目的制备农药醚菊酯类似物和修饰物,与载体蛋白偶联成完全抗原,以之免疫大耳白兔制备抗醚菊酯抗体,为醚菊酯检测试剂的研制做好材料准备。方法利用3-苯氧基苯甲醇和琥珀酸酐合成带羧基的醚菊酯类似物,用重氮法合成对氮基苯乙酸和醚菊酯的偶氮化合物;活化酯法将合成的半抗原与牛血清白蛋白(bovine serum albumin,BSA)及鸡卵清蛋白(ovalbumin,OVA)偶联成完全抗原;并免疫大耳白兔,制备出醚菊酯抗血清,经蛋白G(protein G)柱纯化得到多抗。结果制备的醚菊酯完全抗原偶联比达8∶1和15∶1;制备的多抗效价高于1∶1 280 000;抗体IC_(50)为ng级。结论成功制备了高质量醚菊酯完全抗原及相应抗体,为下一步研制醚菊酯快速检测试剂打下坚实基础。
Objective Etofenprox analogues and modifications were coupled with carrier proteins to form complete antigens which were used to immunize Japanese white rabbits to prepare anti etofenprox antibodies,and to prepare materials for the development of etofenprox detection reagents.Methods Synthesizing etofenprox analogues with carboxyl group by using 3-phenoxybenzyl alcohol and succinic anhydride,and synthesizing azo compounds of p-Nitrophenylacetic acid and etofenprox by a diazo method.The synthesized hapten was coupled with bovine serum albumin(BSA) and ovalbumin(OVA) to form complete antigens by activated ester method.Then the etofenprox antiserum was prepared by immunizing Japanese white rabbits,and purified by protein G column to obtain multi antibodies.Results The results showed that etofenprox complete antigens were prepared with coupling ratios of 8∶1 and 15∶1,the multi antibody titers were higher than 1∶1 280 000 and the antibodies’ IC_(50) was ng grade.Conclusion The high-quality etofenprox complete antigens and corresponding antibodies were successfully prepared,which laid a solid foundation for the further development of the next step of the rapid detection reagent of etofenprox.
作者
吕俊海
杨静
修成奎
王雪
LYU Jun-hai;YANG Jing;XIU Cheng-kui;WANG Xue(Beijing Key Laboratory of TCM Basic Research on Prevention and Treatment of Major Disease,Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing100700,China)
出处
《医学动物防制》
2021年第3期228-232,236,共6页
Journal of Medical Pest Control
基金
中国中医科学院自主课题基金(ZZ2017003)。
关键词
醚菊酯
完全抗原
多克隆抗体
农药
免疫检测
Etofenprox
Complete antigen
Polyclonal antibody
Pesticide
Immune detection
