Abstract
Cow’s milk allergy is defined as an immunologically mediated adverse reaction to cow’s milk proteins and it is usually, along with hen’s egg allergy, the first food allergy identified in childhood.
One of the main aspects to consider when evaluating the allergenic potential of food proteins is the effect of gastric digestion. It is known that allergens are usually able to survive the harsh acidic environment of the stomach, tolerate the presence of surfactants, and resist digestion by pepsin. They might also be digested into high molecular weight peptide fragments, which retain the same, or sometimes increased, IgE-binding. In this respect, western blotting is a highly sensitive and efficient technique that we have used to detect IgE-binding to the digests of milk and egg proteins. Given the importance of the resistance of food proteins to gastric digestion in their capacity to modulate the immune response, we describe in this chapter the assessment of IgE reactivity of a relevant cow’s milk allergen, β-casein, by western blotting after simulated digestion under relevant physiological conditions.
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References
Burks AW, Tang M, Sicherer S et al (2012) ICON: food allergy. J Allergy Clin Immunol 129:906–920
Schoemaker AA, Sprikkelman AB, Grimshaw KE et al (2015) Incidence and natural history of challenge-prov en cow’s milk allergy in European children—EuroPrevall birthcohort. Allergy 70:963–972
Nwaru BI, Hickstein L, Panesar SS et al (2014) The epidemiology of food allergy in Europe: a systematic review and meta-analysis. Allergy 69:62–75
Herz U (2008) Immunological basis and management of food allergy. J Ped Gastroenterol Nutr 47:S54–S57
Restani P, Ballabio C, Di Lorenzo C et al (2009) Molecular aspects of milk allergens and their role in clinical events. Anal Bioanal Chem 395:47–56
Shek LP, Bardina L, Castro R et al (2005) Humoral and cellular responses to cow milk proteins in patients with milk-induced IgE-mediated and non-IgE-mediated disorders. Allergy 60:912–919
Monaci L, Tregoat V, van Hengel AJ et al (2006) Milk allergens, their characteristics and their detection in food: a review. Eur Food Res Technol 223:149–179
Benedé S, López-Expósito I, Molina E et al (2015) Egg proteins as allergens and the effects of the food matrix and processing. Food Funct 6:694–713
Mine Y, Zhang M (2008) Recent advances in the understanding of egg allergens: basic, industrial, and clinical perspectives. J Agric Food Chem 56:4874–4900
Cooke SK, Sampson HA (1997) Allergenic properties of ovomucoid in man. J Immunol 159:2026–2032
Verhoeckx KC, Vissers YM, Baumert JL et al (2015) Food processing and allergenicity. Food Chem Toxicol 80:223–240
Benedé S, López-Expósito I, Giménez G et al (2014a) In vitro digestibility of bovine β-casein with simulated and human oral and gastrointestinal fluids. Identification and IgE-reactivity of the resultant peptides. Food Chem 143:514–521
Benedé S, López-Expósito I, Giménez G et al (2014b) Mapping of IgE epitopes in in vitro gastroduodenal digests of β-lactoglobulin produced with human and simulated fluids. Food Res Int 62:1127–1133
Benedé S, López-Expósito I, López-Fandiño R et al (2014c) Identification of IgE binding peptides in hen egg ovalbumin digested in vitro with human and simulated gastroduodenal fluids. J Agric Food Chem 62:152–158
Benedé S, López-Fandiño R, Reche M et al (2013) Influence of the carbohydrate moieties on the immunoreactivity and digestibility of the egg allergen ovomucoid. PLoS One 8(11):e80810
Jiménez-Saiz R, Benedé S, Miralles B et al (2014) Immunological behavior of in vitro digested egg-white lysozyme. Mol Nutr Food Res 58:614–624
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Benedé, S., López-Fandiño, R., Molina, E. (2017). Assessment of IgE Reactivity of β-Casein by Western Blotting After Digestion with Simulated Gastric Fluid. In: Lin, J., Alcocer, M. (eds) Food Allergens. Methods in Molecular Biology, vol 1592. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6925-8_13
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DOI: https://doi.org/10.1007/978-1-4939-6925-8_13
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