摘要
通过碳二亚胺法将吡虫啉交联于牛血清蛋白(BSA)作为免疫抗原,混合酸酐法将吡虫啉交联于卵清蛋白(OVA)作为包被抗原,免疫Balb/c小鼠,采用B细胞杂交瘤技术,经免疫、融合、筛选、克隆,得到抗吡虫啉单克隆抗体,抗体亚类为IgG1,制备的单克隆抗体效价达1×107,确定了吡虫啉酶联免疫吸附分析方法(ELISA)的最佳工作条件,建立了定量测定吡虫啉的间接竞争ELISA方法。本方法的IC50为(15.12±1.28)μg/L,检出限为(1.76±0.02)μg/L。与其它吡虫啉结构类似物无交叉反应。批内相对标准偏差为4.5%;批间相对标准偏差5.1%,饮用水、重庆理工大学地下水和重庆市花溪河地表水平均添加回收率分别为102%,97%和85%。本研究建立了一种快速检测环境水中吡虫啉残留的方法。
To measure imidacloprid(IMI) in water,the hapten(imidacloprid) was conjugated to bovine serum albumin(BSA) with the carbodiimide method to form immunogen IMI-BSA;the hapten imidacloprid was conjugated to ovalbumin(OVA) with mixed carbonic anhydride method to form the coating antigens.Balb/c mice were immunized intraperitoneally with as prepare immunogen.Hybridoma cell lines secreting monoclonal antibodies(McAbs) against imidacloprid,which was obtained through routine hybridoma technology and ELISA screening.The subtype of secreting McAbs was IgG1.The titer of antibody was 1 × 107.After optimization of the ELISA conditions,an indirect ELISA procedure for the determination of imidacloprid was established.The limit of detection was(1.76 ± 0.02) μg/L and IC50 was(15.12 ± 1.28) μg/L.The McAbs had no cross-reaction with some analogues of imidacloprid.Intra-assay RSD reached 4.5%,and inter-assay RSD reached 5.1%.The recoveries obtained by standard imidacloprid addition to the different samples as drinking water,underwater and surface water were 102%,97% and 85%,respectively.The produced monoclonal antibodies and the developed ELISA procedure may become a convenient and satisfied analytical tool for monitoring imidacloprid residues in environmental samples.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2010年第12期1737-1741,共5页
Chinese Journal of Analytical Chemistry
基金
国家863项目(No.2007AA062419)
重庆市科委自然基金(Nos.2008BB5331
2009BB7231)
重庆市教委自然基金(Nos.KJ090614
KJ100828)
清华大学环境模拟与污染控制国家重点联合实验室专项经费(No.09K05ESPCT)资助项目
关键词
吡虫啉
半抗原
酶联免疫分析法
单克隆抗体
Imidacloprid
Hapten
Enzyme linked immunosorbent assay
Monoclonal antibody
