Comparison of axial and radial flow chromatography on protein separation speed and resolution

Abstract

Fluid dynamic behavior, particularly the relationship between pressure drop and liquid flowrate, inside an axial and a radial flow Chromatographic column packed with compressible porous media was theoretically analyzed using the modified Kozeny-Carman equation. The results were compared with experimental observations obtained using compressible DEAE-agarose as a model medium. At the 2–9 psi pressure drop range studied, theoretical derivation accounting for the gel compression’ effect predicted simple Langmuirian-type response of volumetric flowrate to changes in pressure drop. On the other hand, the experimental result was more or less sigmoidal. At the same pressure drop, radial column yielded 2–3 times higher flowrates than those of axial column both theoretically and experimentally. Using r-HBsAg crude extract, protein resolution effects between the two types of columns at various flowrates were compared side-by-side. Though general Chromatographic behaviors were very similar, the axial column was somewhat superior in terms of rHBsAg recovery yield and specificity. However, the number of theoretical plates analysis indicated the protein resolution effects were comparable.