利用基于单链抗体的BIAcore技术检测蔬菜中的杀螟硫磷残留  

Detection of Residual Fenitrothion in Vegetable Samples by BIAcore Based on Single Chain Antibody

作　　者：刘珊[1] 罗义辉[1] 

机构地区：[1]重庆科技学院化学化工学院,重庆401331

出　　处：《河南农业科学》2017年第5期95-99,共5页Journal of Henan Agricultural Sciences

基　　金：国家质量监督检验检疫总局公益性行业科研专项(2012104003)

摘　　要：为建立一种基于单链抗体的蔬菜中杀螟硫磷残留快速检测方法,测定了前期通过核糖体展示所获得的抗杀螟硫磷单链抗体的亲和力和特异性,并建立了基于该抗体和BIAcore X系统检测蔬菜样品中杀螟硫磷的方法。结果表明,获得3株高亲和力单链抗体,其与杀螟硫磷的解离平衡常数分别为4.56×10^(-10)、1.42×10^(-9)、2.66×10^(-10)mol/L,与除对硫磷外的其他有机磷农药的交叉反应率<0.1%(与对硫磷的交叉反应率≤2.8%),说明获得的单链抗体对杀螟硫磷具有很高的特异性。样本添加回收率试验结果显示,该方法的平均回收率介于95.8%~102.2%,相对标准偏差(RSD)≤2.13%,说明该试验方法可靠,可用于实际蔬菜样品中杀螟硫磷的快速检测。For establishing a method to fast measure residual fenitrothion in vegetables based on single chain antibody,the affinity and specificity of single chain variable fragments( ScFvs) against fenitrothion selected by ribosome display in our preliminary study were tested,and then a method for detection of residual fenitrothion in real vegetable samples by BIAcore X system based on the ScFvs was established.The results showed that three ScFvs of high affinity were obtained,the equilibrium dissociation constants with fenitrothion were 4. 56 × 10^(-10),1. 42 × 10^(-9),2. 66 × 10^(-10)mol/L,respectively,and the cross-reactivity with other organophosphorus pesticides was below 0. 1% except for parathion-methyl( ≤2. 8%),which indicated that the ScFvs had a high specificity for fenitrothion. Recoveries of fenitrothion from fortified vegetables were in the range of 95. 8% —102. 2% and the relative standard deviation( RSD) was below2. 13%. The results above indicated that this method was reliable,and could be used for fast measuring residual fenitrothion in real vegetable samples.

关 键 词：杀螟硫磷 有机磷检测 单链抗体 BIACORE 

分 类 号：S481.8[农业科学—农药学]