基于双特异性单克隆抗体的克百威和三唑磷多残留ELISA分析方法研究及在环境样品中应用  

Development of a Multi-Residue Enzyme-Linked Immunoassay Based on Bispecific Bonoclonal Antibody for Carbufuran and Triazophos and Its Environmental Application

作　　者：金仁耀[1] 朱国念[2] 

机构地区：[1]浙江工商大学食品与生物工程学院,浙江杭州310012 [2]浙江大学农药与环境毒理研究所,浙江杭州310029

出　　处：《核农学报》2013年第4期515-522,531,共9页Journal of Nuclear Agricultural Sciences

摘　　要：采用杂交-杂交瘤技术制备抗克百威和三唑磷的双特异性单克隆抗体并进行纯化,通过对反应模式、工作浓度、包被缓冲液、有机溶剂、离子强度、pH值等条件优选,建立优化的ELISA分析方法,对克百威和三唑磷的线性回归方程分别为Y=20.009ln(x)-9.9293(R2=0.9959)和Y=19.486ln(x)+39.752(R2=0.9945),抑制中浓度I50分别为20ng.mL-1和1.69ng.mL-1,最低检测限I20分别为4.46ng.mL-1和0.36ng.mL-1。建立的ELISA分析方法对水、土壤等环境样品中克百威和三唑磷的平均添加回收率介于88.4%～117%之间,说明所制备的双特异性单克隆抗体和建立的ELISA分析方法可靠,可应用于克百威和三唑磷的多残留免疫分析。The bispecific monoclonal antibody to N-methylcarbamate pesticide carbofuran and Organophosphorus pesticide triazophos was produced by hybrido hybridoma （tetradoma） technology. After purifying the antibody, the optimized ELISA condition, like reaction format, working concentration, coating buffer solution, organic solvent, phosphate ion strength and pH value were investigated. The developed immunoassay showed that the linear recursive equation followed Y =20.009In（x） -9.9293（R^（2） =0.9959 and Y = 19.486In（x） ＋39.752（R^（2） =0.9945） for carbofuran and triazophos, respectively. Inhibition rate analysis showed that 150 and I20 values were 20ng· mL^（-1） ,4.46ng · mL^（-1） and 1.69ng· mL^（-1）, 0.36ng·mL^（-1）for carbofuran and triazophos, respectively. The mean recovery of earbofuran and triazophos in environmental samples such as water and soils were 88.4% and 117% , respectively. The result showed that the developed immunoassay based on bispecific monoclonal antibody was reliable and useful for multiresidue assay for pesticide carbofuran and triazophos.

关 键 词：双特异性单克隆抗体 克百威 三唑磷 多残留检测 ELISA 

分 类 号：S481.8[农业科学—农药学]