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Detection of cows' milk in ewes' milk and cheese by an indirect enzyme-linked immunosorbent assay (ELISA)

Published online by Cambridge University Press:  01 June 2009

Elena Rodríguez ,
Rosario Martín ,
Teresa García ,
Pablo E. Hernández  and
Bernabé Sanz
Elena Rodríguez
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Rosario Martín
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Teresa García
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Pablo E. Hernández
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Bernabé Sanz
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
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Summary

An indirect enzyme-linked immunosorbent assay was successfully developed for the detection of defined amounts of cows' milk (1–50%) in sheeps' milk and cheese. The assay used polyclonal antibodies raised in rabbits against bovine caseins (BC). The anti-BC antibodies were recovered from the crude antiserum by immunoadsorption and elution from a column containing BC. The antibodies were biotinylated and rendered cows' milk specific by mixing them with lyophilized ovine and caprine caseins. ExtrAvidin-peroxidase was used to detect the biotinylated anti-BC antibodies bound to BC immobilized on 96-well plates. Subsequent enzymic conversion of substrate gave clear absorbance differences when assaying mixtures of sheeps' milk and cheese containing variable amounts of cows' milk.

Type
Original Articles
Information
Journal of Dairy Research , Volume 57 , Issue 2 , May 1990 , pp. 197 - 205
Copyright
Copyright © Proprietors of Journal of Dairy Research 1990

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