Immune-analytical detection of the cross-reactive major cereal allergens

Investigation of allergens of celiac disease and cereal allergy has become important for scientific research. Immune-blotting is currently one of the most appropriate methods applied for the identification of potential allergens. The research was focused on the examination of in vitro allergenic activity of proteins in domestic wheat, triticale, rye, barley, rice, corn, buckwheat and amaranth cultivars. The IgA/IgE reactive epitopes of water/salt- and alcohol-soluble protein fractions, respectively, were detected with IgA antibodies of celiac and IgE antibodies of wheat allergic human sera. Further efforts were made to separate the alpha-amylase inhibitors from the water/salt-soluble protein fraction of wheat by DEAE-cellulose anion-exchange chromatography and to confirm their presence on native gel. The most frequently recognised IgE reactive proteins (27) by wheat allergic human sera were identified by liquid chromatography coupled with mass spectroscopy and accession to databank supply. The pepsin resistance of identified wheat allergens was assessed directly on PVDF membrane-blotted protein bands.