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Journal of Dairy Science Vol. 73 No. 6 1489-1493
© 1990 by American Dairy Science Association ®
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Detection of Bovine Milk in Ovine Milk by an Indirect Enzyme-Linked Immunosorbent Assay

Teresa García 1, Rosario Martín 1, Elena Rodríguez 1, Paloma Morales 1, Pablo E. Hernández 1, and Bernabé Sanz 1

1 Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for detection of bovine milk in ovine milk. Polyclonal antibodies were raised in rabbits against bovine whey proteins. Resultant anti-bovine whey protein antibodies were recovered from crude antiserum by immunoadsorption and elution from a column containing bovine whey proteins. Antibodies were biotinylated and rendered specific for bovine milk by mixing them with lyophilized ovine and caprine whey proteins. ExtrAvidin-peroxidase was used to detect the biotinylated anti-bovine whey protein antibodies bound to bovine milk proteins immobilized on 96-well plates. Subsequent enzymic conversion of substrate resulted in discernible differences in optical density between mixtures of ovine milk that contained variable amounts of bovine milk.

Key Words: milk adulteration • enzyme-linked immunosorbent assay • whey proteins

Submitted on June 12, 1989
Accepted on November 27, 1989






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Copyright © 1990 by the American Dairy Science Association ®.