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

J Dairy Res. 1990 May;57(2):197-205. doi: 10.1017/s0022029900026807.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / immunology
  • Binding, Competitive
  • Biotin
  • Caseins / immunology
  • Cattle
  • Cheese / analysis*
  • Enzyme-Linked Immunosorbent Assay
  • Food Contamination / analysis*
  • Milk / analysis*
  • Sheep
  • Species Specificity

Substances

  • Antibodies
  • Caseins
  • Biotin