Development of a Microsphere-Based Immunoassay Authenticating A2 Milk and Species Purity in the Milk Production Chain
2022
Alexander J. W. Elferink | Deborah Entiriwaa | Paolo Bulgarelli | Nathalie G. E. Smits | Jeroen Peters
Processed milk and milk products produced from bovine milk, commonly contain &beta:-casein A1 (&beta:CA1) and &beta:-casein A2 (&beta:CA2). Since the presence of &beta:CA1 is linked to milk intolerance and digestion problems, A2A2 milk, which only contains &beta:CA2, is proposed as a healthier alternative. To support this health claim, the purity of A2A2-milk has to be guaranteed. In the presented study, a multiplex immunoassay, able to distinguish between &beta:CA2 and &beta:CA1, was developed and real-life applicability was shown on raw milk samples from genotyped A1A1, A1A2 and A2A2 cows. Because of its ability to discriminate between &beta:CA2 and &beta:CA1, this newly developed method was able to detect the addition of common bovine A1A2 milk to A2A2 milk, as low as 1%. Besides the detection of A2A2 milk purity, the developed assay can also be implemented as a rapid phenotyping method at dairy farms to replace the more invasive DNA-based screening. Additionally, the developed method was capable of detecting the addition of common bovine milk up to 1% in sheep, goat, buffalo, horse and donkey milk, which conforms to EU recommendations. In conclusion, a newly developed multiplex method capable of reliably detecting the dilution of A2A2 milk of multiple species, with common bovine milk up to 1%, is presented.
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