Protein Band Dispersion in Axial and Radial Flow Chromatography

Abstract

Abstract Radial flow (RF) column configurations have been developed for larger scale production liquid chromatography, with the primary purpose of increasing throughput rates and decreasing trans-bed pressure drops in comparison to conventional axial flow (AF) columns. The RF columns have been quite successful in attaining these two objectives. In this work, we investigated the nature of protein band dispersion in both axial and radial flow chromatography column configurations, utilizing S-200 Sephacryl gel filtration media with bovine serum albumin as the applied protein. The effects of input feed flow rate as well as input feed albumin concentration on the nature of protein band dispersion were studied. Protein band dispersion was quantified by examining the shape of the eluting protein peak and measuring peak height-to-width (HTW) ratio's for the various flow rates and feed concentrations used in the study. Our results indicate that protein dispersion was larger in RF columns as indicated by smaller peak HTW ratio's than those obtained for AF columns, operated at the same flow rates and feed protein concentrations. We also found that moderately increasing the flow rate increased peak HTW ratio's and resulted in sharper peaks for both AF and RF columns.