Portal de Periódicos da CAPES

Development of Enzyme-Linked Immunosorbent Assays for the Insecticide Chlorpyrifos. 1. Monoclonal Antibody Production and Immunoassay Design

1996; American Chemical Society; Volume: 44; Issue: 12 Linguagem: Inglês

10.1021/jf960144q

1520-5118

Juan J. Manclús, Jaime Primo, Ángel Montoya,

Protein purification and stability

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. MAbs showing the highest affinity to chlorpyrifos in homologous assays (I50 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 (I50 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. Keywords: Insecticide; chlorpyrifos; hapten design; monoclonal antibodies; ELISA; analysis