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KMID : 0545120070170061002
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 6 p.1002 ~ p.1009
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Development of Homogeneous Enzyme Immunoassay for the Organophosphorus Insecticide Fenthion: Assessment of the Effects of Protein-Hapten BridgeStructures on Assay Sensitivity
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Kim Bok-Hee
Park Eun-Yong
Lee Yong-Tae
Lee Jung-Hun
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Abstract
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A rapid, convenient homogeneous competitive enzyme immunoassay for estimating the amount of fenthion is described. The assay utilizes glucose-6-phosphate dehydrogenasehapten conjugates that are inhibited in solution by antibodies obtained from bovine serum albumin-hapten conjugates. In order to investigate the effects of bridging group recognition on the sensitivity of dose response characteristics, the bridging groups of varying alkyl chain length were attached at the phosphate position of fenthion. Among the antibodies used, the one obtained from the use of hapten (fenthion analog) with the same bridging group structure that was used in preparing the enzyme-fenthion conjugates showed maximum inhibition (up to 51.8%) in the absence of fenthion. In the presence of fenthion, the activity of the enzyme-hapten conjugate is regained in an amount proportional to the fenthion concentration. Under the optimized condition, the ED50 value for fenthion was 0.809 μg/ml. The assay developed in this study is a rapid effective screening method for fenthion prior to precise analysis.
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KEYWORD
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Organophosphorus insecticide, fenthion, homogeneous competitive enzyme immunoassay, proteinhapten bridge structures, glucose-6-phosphate dehydrogenase
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