title: Global proteomic screening of protein allergens and advanced glycation endproducts in thermally processed peanuts.
abstract: Peanuts (Arachis hypogaea) are the cause of one of the most prevalent food allergies worldwide. Thermal processing (e.g., roasting) of peanuts and peanut-containing foods results in complex chemical reactions that alter structural conformations of peanut proteins, preventing accurate detection of allergens by most immunochemical and targeted screening methodologies. To improve food allergen detection and support more accurate food labeling, traditional methods for peanut protein extraction were modified to include protein denaturants and solubilization agents. Qualitative characterization by SDS-PAGE and Western blot analyses of raw and variably roasted peanut extracts confirmed improvements in total protein recovery and provided evidence for the incorporation of Ara h 1, Ara h 3, and, to a lesser extent, Ara h 2 into high molecular weight protein complexes upon roasting. Relative quantification of allergens in peanut lysates was accomplished by label-free spectral feature (MS1) LC-MS/MS methodologies, by which peanut allergen peptides exhibiting a differential MS response in raw versus roasted peanuts were considered to be candidate targets of thermal modification. Identification of lysine-modified Maillard advanced glycation endproducts (AGE) by LC-MS/MS confirmed the formation of (carboxymethyl)lysine (CML), (carboxyethyl)lysine (CEL), and pyrraline (Pyr) protein modifications on Ara h 1 and Ara h 3 tryptic peptides in roasted peanut varieties. These results suggest that complex processed food matrices require initial analysis by an untargeted LC-MS/MS approach to determine optimum analytes for subsequent targeted allergen analyses.
authors.affiliation: Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration , 5100 Paint Branch Parkway, College Park, Maryland 20740, United States.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Hebling
authors.forename: Melinda A
authors.initials: MA
authors.lastname: McFarland
authors.forename: John H
authors.initials: JH
authors.lastname: Callahan
authors.forename: Mark M
authors.initials: MM
authors.lastname: Ross
mesh_terms: D000485:Allergens / Q000009:adverse effects / Q000032:analysis* / Q000737:chemistry; D052179:Antigens, Plant / Q000009:adverse effects / Q000032:analysis* / Q000737:chemistry; D010367:Arachis / Q000009:adverse effects / Q000737:chemistry*; D005513:Food Inspection / Q000379:methods; D057141:Food, Preserved / Q000009:adverse effects / Q000032:analysis*; D017127:Glycation End Products, Advanced / Q000009:adverse effects / Q000032:analysis* / Q000737:chemistry; D006358:Hot Temperature / Q000009:adverse effects; D006801:Humans; D015416:Maillard Reaction; D009754:Nuts / Q000009:adverse effects / Q000737:chemistry*; D021183:Peanut Hypersensitivity / Q000209:etiology / Q000517:prevention & control*; D010446:Peptide Fragments / Q000009:adverse effects / Q000032:analysis / Q000737:chemistry; D010940:Plant Proteins / Q000009:adverse effects / Q000032:analysis / Q000737:chemistry; D020543:Proteome / Q000009:adverse effects / Q000032:analysis / Q000737:chemistry; D040901:Proteomics / Q000379:methods
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D017127:Glycation End Products, Advanced; D010446:Peptide Fragments; D010940:Plant Proteins; D020543:Proteome