The fractionation and purification of gliadins by hydrophobic interaction chromatography

Abstract

The apolar nature of wheat gliadins has been studied. Apolar properties have been exploited to fractionate components by hydrophobic interaction chromatography and fractions have been analysed further by polyacrylamide gel electrophoresis. Experiments were carried out in pH 5.0 buffer in absence of additives such as urea or dimethylformamide. Gliadin was not retarded by ethyl or butyl agarose but showed a strong affinity for both the phenyl and octyl derivatives. The strength of the hydrophobic interactions was such that aqueous alcohol was required to achieve appreciable desorption. Even so, only two-thirds of the bound gliadin could be eluted with aqueous ethanol alone; complete desorption required the presence of small concentrations of tetramethylammonium chloride. The elution of protein from columns followed the general order 0-, /3-, a-, and y-gliadin, respectively. This procedure provides a simple system for the fractionation of wheat prolamines under mild conditions. Results suggest that a higher degree of resolution may be attainable by further modification of experimental conditions. Interaction of gliadin with nonpolar groups covalently bound to agarose apparently reflects the presence of hydrophobic patches on the surface of the gliadin molecule.

Combined effects of potassium chloride and ethanol as mobile phase modulators on hydrophobic interaction and reversed-phase chromatography of three insulin variants

“…Nonetheless, the hydrophobic interactions between solute and ligand in both HIC and RPC can be manipulated by salts as well as organic solvents. However, few studies have been carried out on the effect of alcohols on HIC [10][11][12][13], and of inorganic salts on RPC [14,15], and those that have been performed were either focused on a certain separation problem [10,11,15], or on phenomena other than retention, such as protein unfolding during chromatography [14], or peak shape and carryover [12].…”

Enhanced Downstream Processing for a Cell-Based Avian Influenza (H5N1) Vaccine

“…To further eliminate remaining proteins (3145 ± 116.05 ng/mL) from the host cells of the primary H5N1 HPAIV purification solution, we chose Polar MC-HIC, using the hydrophobic nature of phenyl over butyl [ 44 ]. This choice facilitated the ongoing isolation and purification of the viral fluid previously processed by Capto TM Core 700.…”

Rheology of Wheat Doughs for Fresh Pasta Production: Influence of Semolina‐flour Blends and Salt Content

“…It is well established that several factors are responsible for the development of the gluten network: disulphide bonds (Ewart 1985). hydrogen and ionic bonds (Bloksma and Bushuk 1988), Van der Waals and hydrophobic interactions (Caldwell 1979;Huebner and Wall 1980). The use of several compounds as improvers of the rheological properties of dough is based on these intermolecular interactions occurring in the gluten.…”