Among wheat storage proteins, omega-gliadins display a singular amino acid sequence only composed by re-petitive sequences. They have been described as a major wheat allergen and also contain T-cell epitopesimplicated in coeliac disease. To study the structural and biophysical properties of omega-gliadins and othergliadins (e.g., gluten network formation, allergic response), highly purified and concentrated fractions areneeded. In the present work, we used chromatography media screening to improve their fractionation. A SCeramic Hyper D (Pall) resin was selected for its ability to concentrate omega-gliadins in the first elution peaks.Gamma- and beta-gliadins were then separated by hydrophobic interaction chromatography and eluted with anethanol gradient. Alpha-gliadins were purified by gel filtration. Each fraction was characterized by electro-phoresis and reverse phase HPLC. The developed preparative protocol enabled the purification of several gramsof highly purified gliadins to the detriment of the yield.