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Rapid, sensitive two-site ELISA for detection of cows' milk in goats' or ewes' milk using monoclonal antibodies

Published online by Cambridge University Press:  01 June 2009

Didier Levieux  and
Annie Venien
Didier Levieux
Affiliation:
Unité d'Immunochimie, Station de Recherches sur la Viande INRA, Theix, 63122 Saint-Genès-Champanelle, France
Annie Venien
Affiliation:
Unité d'Immunochimie, Station de Recherches sur la Viande INRA, Theix, 63122 Saint-Genès-Champanelle, France
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Summary

A sandwich ELISA (enzyme-linked immunosorbent assay) of the two-site type has been successfully developed for the detection of cows' milk in goats' or ewes' milk. The assay uses two monoclonal antibodies (MAb) raised in mice against cows' β-lactoglobulin (β-lg). These MAb recognize different epitopes of the β-lg, which are sufficiently distinct to allow simultaneous binding of the corresponding antibodies. One of the MAb recognizes a species-specific epitope of the bovine β-lg and was adsorbed to a plastic microtitration plate (capture antibody). The second MAb was labelled with peroxidase and used to detect the captured cows' β-lg. Factors affecting assay performance were investigated. The optimized assay is highly specific, reproducible (intra- and inter-assay CV were 8 and 13% respectively) and sensitive: as little as 5 ng β-lg/ml or 1 part cows' milk per 100000 parts goats' or ewes' milk can be detected. The technique is robust, cheap, rapid, reliable and suitable for high sample throughput, semi-automation and screening surveys. The MAb used guarantee the high specificity of the assay and indefinite reagent supply of constant quality once approved by collaborative national or international trials.

Type
Original Articles
Information
Journal of Dairy Research , Volume 61 , Issue 1 , February 1994 , pp. 91 - 99
Copyright
Copyright © Proprietors of Journal of Dairy Research 1994

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