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Development of an Enzyme-Linked Immunosorbent Assay for 3,5,6-Trichloro-2-pyridinol. 1. Production and Characterization of Monoclonal Antibodies

1996; American Chemical Society; Volume: 44; Issue: 11 Linguagem: Inglês

10.1021/jf950656p

1520-5118

Juan J. Manclús, Ángel Montoya,

Pharmacological Effects and Assays

3,5,6-Trichloro-2-pyridinol (TCP) is the major degradation product of both the widely used chlorpyrifos and chlorpyrifos-methyl insecticides and the herbicide triclopyr. In this paper we describe the development of immunoassays for the detection of this metabolite. With this aim, two types of haptens were synthesized, with spacer attachment either through the 6-position of the pyridyl ring or through the hydroxyl group. Protein conjugates of these haptens were used for the immunization of mice to produce monoclonal antibodies (MAbs). From haptens of the former type, a panel of MAbs displaying 50% inhibition (I50) for TCP in the range 1−10 nM was obtained. Characterization of these MAbs showed their high affinity and specificity for TCP. The best MAbs were used in various assay formats in combination with homologous and heterologous assay haptens. Several immunoassays with I50 lower than 1 nM were developed. These immunoassays are potentially very valuable analytical tools for the determination of TCP in food and in the environment, as well as for monitoring human exposure to chlorpyrifos. Keywords: 3,5,6-Trichloro-2-pyridinol (TCP); chlorpyrifos; monoclonal antibodies; ELISA