环糊精及肽配体介导的毒死蜱非竞争检测模式

摘要

以毒死蜱作为对象,构建了一种由环糊精和肽配体介导的非竞争检测模式.首先利用分子模拟和光谱学方法对β-环糊精包合毒死蜱的效应进行了证实;然后将β-环糊精与牛血清蛋白进行偶联,并证实偶联状态的环糊精分子仍能包合毒死蜱;以β-环糊精-牛血清蛋白偶联产物为核心材料,从商品化的噬菌体展示线性十二肽库中筛选获得了5个可以特异识别环糊精-毒死蜱包合物的肽配体;最后,利用亲和力最强的肽配体(8#克隆)构建毒死蜱的非竞争检测模式,该检测模式能够特异性地识别毒死蜱,而对丙溴磷、除线磷和溴硫磷等结构类似农药分子无识别.本研究所构建的检测模式无需对小分子物质进行改造和固定,可为小分子化学污染物的快速检测提供新的参考.%In this work,a cyclodextrin and peptide ligand-assisted noncompetitive detection model was developed using chlorpyrifos (CPF) as typical target.The possibility of CPF encapsulated by β-cyclodextrin (β-CD) was first confirmed through the use of molecular modeling and UV and fluorescence spectroscopy.Then,bovine serum albumin (BSA)-β-CD conjugate,which was still able to include CPF,was synthesized and five positive peptide ligands specifically bound to the CPF:β-CD inclusion complexes rather other β-CD were screened from a commercialized phage-display liner dodecapeptide library.At last,a noncompetitive detection model was developed using BSA-β-CD conjugate and the most active peptide (Clone 8).The developed detection model could specially detect CPF,but not profenofos,dichlofenthion,or bromophos without the need for the modification and immobilization of the target small molecules,thereby providing an alternative method for the rapid detection of small molecules.