Product(s) used in this publication: PepStar™ Peptide Microarrays
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-caseinvariants B and C, which are frequent in most breeds, starting from milk with identification and purification of caseinvariants by isoelectric focusing (IEF) and anion-exchange chromatography, followed by in vitro gastrointestinal digestion of the caseinvariants, 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 microarrayimmunoassay with cow's milk allergic human sera.
Caseinvariants; IgE epitope mapping; In vitro gastrointestinal digestion; Milk allergy; Peptidemicroarray