Combining 2-DE immunoblots and mass spectrometry to identify putative soybean (Glycine max) allergens. (June 2018)

• Record Type: Journal Article
• Authors: Lu, Mei; Jin, Yuan; Cerny, Ron; Ballmer-Weber, Barbara; Goodman, Richard E.
• Is Part Of: Food and chemical toxicology. Volume 116:Part B(2018)
• Page Start: 207
• Page End: 215
• Publication Date: 2018-06
• Subjects: β-conglycinin -- Glycinin -- IgE -- LC-MS/MS -- Soybean allergens -- Two-dimensional immunoblots; CHAPS 3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate -- DBPCFC double-blind placebo-control food challenge -- GE genetically engineered -- LEP late embryogenesis protein -- MOWSE molecular weight search -- MW molecular weight -- NCBI National Center for Biotechnology -- OECD Organization of Economic Co-operation and Development -- SBP seed biotinylated protein -- SKTI soybean Kunitz trypsin inhibitor -- SPT skin prick test -- vdc volts of direct current -- WHO/IUIS World Health Organization and International Union of Immunological Societies; Toxicology -- Periodicals; Food poisoning -- Periodicals; Food Poisoning -- Periodicals; Toxicology -- Periodicals; Toxicologie -- Périodiques; Intoxications alimentaires -- Périodiques; Food poisoning; Toxicology; Periodicals; Electronic journals; 615.9
• DOI: 10.1016/j.fct.2018.04.032
• Languages: English
• ISSNs: 0278-6915

Abstract

Abstract: Soybean is recognized as a commonly allergenic food, but the identity of important allergens is not well studied. Recently, some global regulatory agencies started requiring quantitative analysis of individual allergens, including unproven allergens, as part of the risk assessment for genetically engineered (GE) soybeans. We sought to identify soybean proteins that bind IgE from any of 10 individual soybean-sensitized subjects. Soybean IgE binding proteins were identified by 2-DE immunoblots using sera from four soy-allergic and plasma from six soy-sensitized human subjects. Corresponding spots were excised from stained gels, digested, and analyzed using a quadrupole TOF Synapt G2-S tandem mass spectrometer. Results showed the major IgE binding proteins were subunits of either β-conglycinin (Gly m 5) or glycinin (Gly m 6). Soybean Kunitz trypsin inhibitor (SKTI) was a significant IgE binding protein for four subjects. Soybean agglutinin, seed biotinylated protein (SBP) of 65 kDa, late embryogenesis protein (LEP), and sucrose-binding protein were identified as IgE binding only for soy-sensitized subjects. We conclude that the major soybean allergens are isoforms of Gly m 5, Gly m 6, and possibly SKTI and that requirements for quantitative measurement of proteins that are not clear allergens is not relevant to safety. Graphical abstract: Highlights: Subunits of β-conglycinin (Gly m 5) and glycinin (Gly m 6) were major soybean allergens. Soybean Kunitz trypsin inhibitor (SKTI) was a significant IgE binding protein in four out of ten subjects. Soy agglutinin, seed biotinylated, late embryogenesis and sucrose-binding proteins bound IgE of soy-sensitized individuals. Many putative soybean allergens were not detected as IgE binding proteins among the ten subjects.