Fast biosensor immunoassays for the detection of cows' milk in the milk of ewes and goats

The Journal of Dairy Research
Willem HaasnootMaria G E G Bremer

Abstract

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).

Citations

Mar 6, 2009·Analytical and Bioanalytical Chemistry·Maria G E G BremerWillem Haasnoot
May 29, 2007·Analytical and Bioanalytical Chemistry·Arjon J van Hengel
Oct 28, 2005·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Dec 24, 2010·Bipolar Disorders·Emily G SeveranceRobert H Yolken
May 16, 2015·Critical Reviews in Food Science and Nutrition·Shyam Narayan JhaRishi Bhardwaj
Mar 23, 2015·Analytical and Bioanalytical Chemistry·Μichailia AngelopoulouSotirios E Kakabakos
Aug 10, 2016·Biosensors & Bioelectronics·J AshleyI E Tothill
Sep 22, 2009·The Analyst·Asta Kausaite-MinkstimieneArunas Ramanavicius
Sep 22, 2010·Analytical Chemistry·Sabina Rebe RazMaria G E G Bremer
Jul 13, 2018·Analytical Chemistry·Michailia AngelopoulouSotirios E Kakabakos

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