title: Effects of processing on the recovery of food allergens from a model dark chocolate matrix.
abstract: To alleviate the risk to allergic consumers, it is crucial to improve factors affecting the detection of food allergens in processed chocolate products. This study evaluated processing effects on (1) recovery of peanut, egg, and milk allergens using five different extraction buffers, and (2) identification of specific allergenic proteins from extracts of incurred chocolate using allergen-specific antibodies and human allergic sera. Immunochemical staining with polyclonal antibodies showed that the addition of detergent or reducing agent improved extraction efficiency of peanut proteins, but not of egg and milk proteins. Tempering decreased antibody binding regardless of extractant. Detection of IgE-reactive peanut, egg, and milk allergens was differentially affected by tempering and extractant. Detection problems associated with matrix and processing effects may be overcome by the choice of extraction buffer and detecting antibody.
authors.affiliation: Center for Food Safety and Applied Nutrition, United States Food and Drug Administration, 8301 Muirkirk rd., Laurel, MD 20708, United States. Electronic address: sefat.khuda@fda.hhs.gov.
authors.forename: Sefat E
authors.initials: SE
authors.lastname: Khuda
authors.affiliation: Center for Food Safety and Applied Nutrition, United States Food and Drug Administration, 6502 South Archer Road, Bedford Park, IL 60501, United States.
authors.forename: Lauren S
authors.initials: LS
authors.lastname: Jackson
authors.affiliation: Center for Food Safety and Applied Nutrition, United States Food and Drug Administration, 6502 South Archer Road, Bedford Park, IL 60501, United States.
authors.forename: Tong-Jen
authors.initials: TJ
authors.lastname: Fu
authors.affiliation: Center for Food Safety and Applied Nutrition, United States Food and Drug Administration, 8301 Muirkirk rd., Laurel, MD 20708, United States.
authors.forename: Kristina M
authors.initials: KM
authors.lastname: Williams
mesh_terms: D000485:Allergens / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D010367:Arachis / Q000378:metabolism*; D015153:Blotting, Western; D002099:Cacao / Q000378:metabolism*; D002645:Chickens; D004529:Egg White / Q000032:analysis; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D008892:Milk / Q000378:metabolism; D008894:Milk Proteins / Q000032:analysis / Q000276:immunology
chemical_list: D000485:Allergens; D000906:Antibodies; D008894:Milk Proteins; D007073:Immunoglobulin E
keywords: Allergenic proteins; Antibody specificity; Chocolate matrix; Extraction buffers; Food allergens; Immunochemical staining; Processing effects