Purification of wheat flour high-molecular-weight glutenin subunits by fast protein liquid chromatography

Fast protein liquid chromatography has been developed for purification of high-molecular-weight glutenin subunits HMW-GSs from wheat flour. Flour samples from four wheat cultivars with different HMW-GS alleles at Glu-A1, Glu-B1 and Glu-D1 loci were used to establish the method. The column material used was Resource™ Phe, and the optimal elution was with a gradient formed with buffer A [0.05 M Tris–HCl containing 4 M urea and 0.25 M (NH4)2SO4, pH 8.0] and buffer B [0.05 M Tris–HCl containing 4 M urea (pH 8.0)] at a flow rate of 0.5 ml/min. A pure single 1Dx-, 1Bx- HMW-GS, and all the y-type HMW-GSs present in one genotype can be reliably separated in a single step.