Wheat Allergens MALDI-TOF/TOF - Using 7 cm immobilized pH gradient strips to determine levels of clinically relevant proteins in wheat grain extracts

The aim of the work was to test relatively simple proteomics approach based on phenol extraction and two-dimensional gel electrophoresis (2-DE) with 7cm immobilized pH gradient strips for the determination of clinically relevant proteins in wheat grain. Using this approach, 157 2-DE spots were quantified in biological triplicate, out of which 55 were identified by MALDI-TOF/TOF mass spectrometry. Clinically relevant proteins associated with celiac disease, wheat dependent exercise induced anaphylaxis, baker´s asthma, and food allergy, were detected in 24 2-DE spots. However, alcohol-soluble gliadins were not detected with this approach. The comparison with recent quantitative study suggested that gel-based and gel-free proteomics approaches are complementary for the detection and quantification of clinically relevant proteins in wheat grain.