Assessment of IgE Reactivity of β-Casein by Western Blotting After Digestion with Simulated Gastric Fluid.
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.
Instituto de Investigación en Ciencias de la Alimentación (CIAL, CSIC-UAM), Nicolás Cabrera 9, Madrid, 28049, Spain. sarabenede@hotmail.com.|Pediatric Allergy and Immunology, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY, 10029, USA. sarabenede@hotmail.com.
Sara
S
Benedé
Instituto de Investigación en Ciencias de la Alimentación (CIAL, CSIC-UAM), Nicolás Cabrera 9, Madrid, 28049, Spain.
Rosina
R
López-Fandiño
Instituto de Investigación en Ciencias de la Alimentación (CIAL, CSIC-UAM), Nicolás Cabrera 9, Madrid, 28049, Spain.
Elena
E
Molina
D000485:Allergens / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002364:Caseins / Q000378:metabolism*; D002417:Cattle; D021181:Egg Hypersensitivity / Q000378:metabolism; D004531:Eggs; D005512:Food Hypersensitivity / Q000378:metabolism; D005744:Gastric Acid / Q000378:metabolism*; D007073:Immunoglobulin E / Q000378:metabolism*; D008892:Milk / Q000378:metabolism; D016269:Milk Hypersensitivity / Q000378:metabolism
D000485:Allergens; D002364:Caseins; D007073:Immunoglobulin E
Digestibility; Food allergy; Gastric digestion; Immunoassay; SDS-PAGE; Simulated gastric fluid; Western blotting