HPCPC separation of proteins using polyethylene glycol-potassium phosphate aqueous two-phase.

ABSTRACT

High Performance Centrifugal Partition Chromatography (HPCPC) is a practical and suitable method, particularly on the preparative scale, for the separation of biomolecules such as proteins, enzymes, etc. Aqueous two-phase system is also very attractive for the isolation of biomolecules. Aqueous polymer phase system composed of polyethylene glycol 6000-potassium phosphate has been used for the countercurrent chromatographic separation of bovine serum albumin (BSA) and lysozyme using HPCPC. The separation of BSA and lysozyme under various conditions such as various flow rates, rotational speeds, pH of the solvent, and the retention of stationary phase has been studied in the present investigation. The baseline separation of BSA and lysozyme has been also observed. The results of this study demonstrate that HPCPC is useful for separation of proteins with aqueous two-phase systems.