PROTEIN SEPARATION BY CROSS-AXIS COIL PLANET CENTRIFUGE WITH TWO DIFFERENT POSITIONS OF ECCENTRIC COIL ASSEMBLIES USING POLYETHYLENE GLYCOL-DEXTRAN SOLVENT SYSTEM

Abstract

Countercurrent chromatography (CCC) of proteins was performed by the cross-axis coil planet centrifuge (cross-axis CPC) using polyethylene glycol (PEG) - dextran biphasic systems. A pair of eccentric coil assemblies was mounted at two different positions (off-center and central) on the rotary frame. The best separation was obtained from a two-phase solvent system composed of 4.0% (w/w) PEG 8000 - 5.0% (w/w) dextran T500 in 5 mM potassium phosphate buffer (pH 7.0) and 3 M sodium chloride using the upper PEG-rich phase as the mobile phase. The off-center column position yielded substantially better peak resolution between lysozyme and myoglobin than that obtained from the central position under otherwise identical conditions.

ACKNOWLEDGMENT

The authors wish to thank Mr. Tomohiro Miura (Division of Physical Chemistry, College of Pharmacy, Nihon University, Chiba, Japan) for his helpful advice.