APPLICATION OF TWO-DIMENSIONAL ELECTROPHORESIS AND MASS SPECTROMETRY FOR THE DETECTION OF ALLERGENS IN SELECTED VARIETIES OF WHEAT, OATS AND BUCKWHEAT

Abstract

The number of people suffering from food allergies and intolerances has been increasing in recent years and cereal proteins are the most common food allergens. Pseudocereals represent perspective alternative in nutrition with a positive effect on the human body. The aim of the work was to analyze the proteome of selected varieties of wheat (Triticum aestivum L.), oats (Avena sativa) and buckwheat (Fagopyrum esculentum Moench.) using two-dimensional electrophoresis (2DE) and mass spectrometry in order to detect the presence of potentially allergenic proteins. Using the PDQuest program, 221 protein spots ranging from 4.13 to 9.89 µl with experimental molecular weights from 12.42 kDa to 140 kDa were quantified in 2DE gels of wheat. In the oat sample, 168 protein spots were quantified in the range pI of 4.02 to 9.93 and an experimental molecular weight of 14.81 kDa to 67.96 kDa. Buckwheat proteins were separated on a 2DE gel into 208 protein spots in the 3 to 9.83 pI region with an experimental molecular weight of 10.10 kDa to 115 kDa. By comparing the data with the Allergome database, allergens Tri and 26, Tri and 33, Tri and 36, Tri and alpha Gliadin, Tri and 20 were detected in wheat, Ave s 11S allergens in oats and Fag e 1 allergen in buckwheat. 2DE together with mass spectrometry have been shown to be suitable and sensitive methods for the detection of allergens in food crops.