Immunoglobulin E epitope mapping of milk proteins reveals important information about their immunologic properties. Genetic variants of αS1-casein, one of the major allergens in bovine milk, are until now not considered when discussing the allergenic potential. Here we describe the complete procedure to assess the allergenicity of αS1-casein variants B and C, which are frequent in most breeds, starting from milk with identification and purification of casein variants by isoelectric focusing (IEF) and anion-exchange chromatography, followed by in vitro gastrointestinal digestion of the casein variants, identification of the resulting peptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDITOF- MS), in silico analysis of the variant-specific peptides as allergenic epitopes, and determination of their IgE-binding properties by microarray immunoassay with cow’s milk allergic human sera.
CITATION STYLE
Lisson, M., & Erhardt, G. (2016). Mapping of epitopes occurring in bovine αs1-casein variants by peptide microarray immunoassay. In Methods in Molecular Biology (Vol. 1352, pp. 279–296). Humana Press Inc. https://doi.org/10.1007/978-1-4939-3037-1_21
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