Purification of soybean cupins and comparison of IgE binding with peanut allergens in a population of allergic subjects

• Published in: Food and chemical toxicology Vol. 147; p. 111866
• Main Authors: Ramadan, Samah, Marsh, Justin, El-Sherbeny, Ghada A., El-Halawany, El-Sayed F., Luan, Fulei, Baumert, Joseph L., Johnson, Philip, Osman, Yehia, Goodman, Richard E.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.01.2021
• Subjects: albumins; Amino Acid Sequence; Antigens, Plant - chemistry; Antigens, Plant - immunology; Ara h 1; Ara h 2; Ara h 3; Ara h 6; Arachis - chemistry; Chromatography, Liquid; cross reaction; Cross Reactions; Globulins - chemistry; Gly m 5; Gly m 6; Glycine max - chemistry; Humans; IgE immunoblotting; immunoblotting; immunogenicity; Immunoglobulin E; LC-MS/MS; liquid chromatography; Peanut Hypersensitivity; peanuts; Protein Binding; Seed Storage Proteins - chemistry; Soybean Proteins - chemistry; soybeans; Tandem Mass Spectrometry; toxicology; vicilin; LC-MS/MS; Gly m 6; Gly m 5; Ara h 6; Ara h 1; Ara h 2; Ara h 3; IgE immunoblotting
• ISSN: 0278-6915; 1873-6351; 1873-6351
• DOI: 10.1016/j.fct.2020.111866

Identification, purification and characterization of allergens is crucial to the understanding of IgE-mediated disease. Immunologic and structural studies with purified allergens is essential for understanding relative immunogenicity and cross-reactivity. In this work, the complex soybean 7S vicilins (Gly m 5) with three subunits and 11S legumins (Gly m 6) with five subunits were purified and characterized along with purified peanut allergens (Ara h 1, 2, 3, and 6) by label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). Individual subjects plasma IgE binding was tested from subjects allergic to soybeans and or peanuts by immunoblotting, ImmunoCAP™ and ISAC™ ImmunoCAP chip, comparing these soybean proteins with those of purified peanut allergens; vicilin (Ara h 1), 2S albumin (Ara h 2 and Ara h 6) and 11S globulin (Ara h 3). Results show differences between methods and subjects demonstrating the complexity of finding answers to questions of cross-reactivity.