Chromatography of hen egg-white proteins on anion-exchange cellulose at high flow rates using a radial flow column.

Abstract

The influence of column configuration on the separation of hen egg-white proteins using Whatman DE52 and QA52 anion-exchange cellulose has been investigated. Using a 100 ml volume axial flow column (6.6 cm x 4.4 cm i.d.) we achieved flow rates of up to 25 ml/min i.e. 15 bed volumes/h after which higher flow was restricted due to pressure constraints within the system. Under radial flow conditions using a 100 ml column flow rates of up to 150 ml/min i.e. 90 bed volumes/h were achieved using DE52 and QA52. While chromatographic resolution was superior under axial flow at the lower flow rates excellent resolution was maintained at up to 150 ml/min using the radial flow column. This is a consequence of the fast kinetics of adsorption/desorption exhibited by DE52 and QA52. The data indicate that it is the column configuration and not the cellulose matrix which influences flow performance.