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Mapping of Epitopes Occurring in Bovine α s1-Casein Variants by Peptide Microarray Immunoassay

Lisson et al., J Dairy Sci. (2014) - PMID: 26490483

Product(s) used in this publication:  PepStar™ Peptide Microarrays

Abstract:

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 (MALDI-TOF-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.

KEYWORDS:

Casein variants; IgE epitope mapping; In vitro gastrointestinal digestion; Milk allergy; Peptide microarray

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