Mapping of Epitopes Occurring in Bovine α(s1)-Casein Variants by Peptide Microarray Immunoassay.
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.
Department of Animal Breeding and Genetics, Justus-Liebig University, Ludwigstraße 21 B, Gießen, 35390, Germany.
Maria
M
Lisson
Department of Animal Breeding and Genetics, Justus-Liebig University, Ludwigstraße 21 B, Gießen, 35390, Germany. Georg.Erhardt@agrar.uni-giessen.de.
Georg
G
Erhardt
D019943:Amino Acid Substitution; D000818:Animals; D002364:Caseins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D002417:Cattle; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D004386:Duodenum / Q000378:metabolism; D018604:Epitope Mapping / Q000379:methods; D005753:Gastric Mucosa / Q000378:metabolism; D014644:Genetic Variation; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007073:Immunoglobulin E / Q000276:immunology; D007525:Isoelectric Focusing; D008892:Milk / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D040081:Protein Array Analysis / Q000379:methods*; D059748:Proteolysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D002364:Caseins; D010446:Peptide Fragments; D007073:Immunoglobulin E
Casein variants; IgE epitope mapping; In vitro gastrointestinal digestion; Milk allergy; Peptide microarray