Chapter title |
Mapping of Epitopes Occurring in Bovine αs1-Casein Variants by Peptide Microarray Immunoassay.
|
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Chapter number | 21 |
Book title |
Peptide Microarrays
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Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3037-1_21 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3036-4, 978-1-4939-3037-1
|
Authors |
Maria Lisson, Georg Erhardt |
Editors |
Marina Cretich, Marcella Chiari |
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. |
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