Development of enzyme-linked immunosorbent assays for the insecticide chlorpyrifos .1. Monoclonal antibody production and immunoassay design

被引:67
作者
Manclus, JJ
Primo, J
Montoya, A
机构
[1] UNIV POLITECN VALENCIA,CSIC,LAB INTEGRADO BIOINGN,VALENCIA 46022,SPAIN
[2] UNIV POLITECN VALENCIA,CSIC,INST TECNOL QUIM,VALENCIA 46022,SPAIN
关键词
insecticide; chlorpyrifos; hapten design; monoclonal antibodies; ELISA; analysis;
D O I
10.1021/jf960144q
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
The present work describes the production and characterization of monoclonal antibodies (MAbs) to the insecticide chlorpyrifos and their incorporation into several ELISA configurations. With this aim, a collection of chlorpyrifos haptens was synthesized by introducing appropriate spacers in opposite sites of the analyte molecular structure. From mice immunized with protein conjugates of these haptens, several hybridomas secreting MAbs with the ability to sensitively bind the analyte were obtained. IMAbs showing the highest affinity to chlorpyrifos in homologous assays (I-50 values in the 20-220 nM range) were selected. Hapten heterology involving modifications of the moiety closer to the attachment site provided the highest improvement in sensitivity. MAbs displayed striking differences in their cross-reactivity pattern with structurally related compounds. One MAb (I-50 around 10 nM) was incorporated into other ELISA formats. No remarkable changes of assay characteristics, other than immunoreagent consumption and immunoassay procedure, were found. These ELISAs are potentially very valuable analytical tools for the rapid and sensitive determination of this insecticide.
引用
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页码:4052 / 4062
页数:11
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