MEMBRANE CHROMATOGRAPHIC SYSTEMS FOR HIGH-THROUGHPUT PROTEIN SEPARATIONS

This paper explores the utility of a membrane chromatographic system (MemSep) for analytical and preparative separations of biomolecules. These column systems consist of stacked disks of macroporous cross-linked regenerated cellulose membranes functionalized with ion-exchange moieties. Fluid flow through the macropores of these membranes results in rapid mass transport to and from the adsorbent surface. Elution and frontal experiments demonstrated that these systems were relatively insensitive to flow-rate. Linear gradient experiments under analytical conditions indicated that rapid separations could be readily carried out. Preparative-scale separations of proteins on ion-exchange MemSep systems were scaled-up with respect to flow-rate and mass loading with minimal adverse effect on bioproduct purity. A cation-exchange CM MemSep 1010 device was able to concentrate and purify 30 mg and 15 mg of proteins in 3 min when operated in the step and linear gradient modes, respectively. The design of these membrane chromatographic systems enables efficient gradient elution of proteins under elevated flow-rate and mass loading conditions.