Fast biosensor immunoassays for the detection of cows' milk in the milk of ewes and goats.
Two monoclonal antibodies (MAb) raised against bovine kappa-casein were developed and applied in an automated optical biosensor (Biacore 3000) to create easy and fast direct and inhibition biosensor immunoassays (BIA) for the detection of cows' milk in the milk of ewes and goats. With both assay formats, low limits of detection (<01%) and fast run times (around 5 min) were obtained. For sample preparation, milk was diluted in buffer (direct assay) or in an antibody-containing buffer (inhibition assay) only. For quantitative analysis, calibrants of cows' milk in ewes' or goats' milk were used. Advantages of the direct BIA are: the single reagent format (biosensor chip immobilized antibodies only); the use of small amounts of antibodies (2 microg for >350 tests); and the wide measurement range (0.1 to 10% cows' milk). Despite these advantages, the inhibition BIA (using kappa-casein immobilized on the chip) was preferred because of the possible application of non-purified Mab, the higher responses, the higher sensitivity at relevant low percentages of cows' milk and its robustness (>800 cycles per chip).
Haasnoot
Willem
W
RIKILT-Institute of Food Safety, PO Box 230, 6700 AE Wageningen, The Netherlands. willem.haasnoot@wur.nl
Smits
Nathalie G E
NG
Kemmers-Voncken
Anniek E M
AE
Bremer
Maria G E G
MG
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D015374:Biosensing Techniques; D002364:Caseins / Q000032:analysis / Q000276:immunology; D002417:Cattle; D000939:Epitopes / Q000276:immunology; D005260:Female; D005506:Food Contamination / Q000032:analysis; D006041:Goats; D007118:Immunoassay / Q000379:methods / Q000662:veterinary*; D007202:Indicators and Reagents; D008892:Milk / Q000737:chemistry / Q000145:classification*; D008894:Milk Proteins / Q000032:analysis; D012680:Sensitivity and Specificity; D012756:Sheep; D012996:Solutions; D013997:Time Factors
D000911:Antibodies, Monoclonal; D002364:Caseins; D000939:Epitopes; D007202:Indicators and Reagents; D008894:Milk Proteins; D012996:Solutions