Soybean Allergens Affecting North American Patients Identified by 2D Gels and Mass Spectrometry
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Soybean Allergens Affecting North American Patients Identified by 2D Gels and Mass Spectrometry Christine Gagnon & Vaino Poysa & Elroy R. Cober & Stephen Gleddie
Received: 1 December 2008 / Accepted: 20 May 2009 / Published online: 26 June 2009 # Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada 2009
Abstract In this study, we compared the immunoglobulin E immunological reactions of 23 soy-allergic, nine soysensitive, and four non-allergic human sera to soybean proteins separated by 1D and 2D gel electrophoresis and screened by Western blot. This method led to novel allergen identification in soybean proteins by tandem mass spectrometry analysis of reactive proteins. Soybean is one of the eight most significant foods which provoke allergic reactions among North Americans. Previous studies have identified several putative allergens present in soybean seeds; however, many of these reports did not employ mass spectrometry to conclusively identify the allergenic proteins. Reviews have suggested that soybean seeds may contain between 10 and 20 different proteins responsible for provoking allergic reactions among sensitive patients. We report, in this study of North American patients, a total of 19 potentially allergenic proteins including ten identified by mass spectrometry and five novel allergens. We have also made extensive use of soybean lines lacking various subunits of the major seed storage proteins, glycinin and βconglycinin. By using these knockout lines in Western blots with patient serum, it was conclusively demonstrated that some patients react predominantly to only a few proteins, while most react to four to six proteins. The findings herein describe the main allergic proteins present in soybean C. Gagnon : E. R. Cober : S. Gleddie (*) Eastern Cereal and Oilseed Research Centre, Central Experimental Farm, Agriculture and Agri-Food Canada, Ottawa, Ontario K1A 0C6, Canada e-mail: [email protected] V. Poysa Greenhouse and Processing Crops Research Centre, Agriculture and Agri-Food Canada, Harrow, Ontario N0R 1G0, Canada
seeds, the relative significance of these allergens among North Americans, and some genetic lines of soybean lacking individual allergens. Keywords Soybean Allergy . Soy-Allergic Human Serum . Soy-Sensitive Human Serum . 2D Gel Electrophoresis . IgE Western Blot . Tandem Mass Spectrometry
Introduction Soybeans (Glycine max L. Merr.) are an excellent source of high-quality protein, polyunsaturated fats, vitamins, minerals, and other nutrients for both human food and animal feed (Krishnan 2000; Wang et al. 2003). Soy proteins are added to a wide range of food products, such as infant formulas, meal replacement drinks, and sports bars and beverages. As well as increasing the nutritional value of the food, the US Food and Drug Administration and the Joint Health Claims Initiative in the UK, among other regulatory bodies, have established that adding soy proteins to food reduces cholesterol levels, thereby lowering the risk of cardio
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