HIGH PRESSURE GEL PERMEATION CHROMATOGRAPHY OF NATIVE PROTEINS ON TSK‐SW COLUMNS

Abstract

Twenty-five proteins in their native state as well as additional materials for the evaluation of column parameters have been submitted to high pressure gel permeation chromatography (GPC) on a TSK G3000 SW column. Our data are compiled from 110 experiments. A new method of treating GPC data has been developed and compared with other methods currently in use. The new method yields lower errors in molecular weights than the others, and by comparison with theoretical plots we were able to attribute the larger errors to curvature of the alternative plots. After studying several methods for characterizing the column, we have concluded that thyroglobulin as well as three or four proteins in the molecular weight range 20,000–300,000, all containing sodium azide, serve to characterize the column with respect to V₀, V_iand V_t. Extension of this method of analysis to denatured proteins, biopolymers and other polymers is also out lined. Several chromatograms are also presented to illustrate the resolution obtained with columns of this type. Column parameters are compared with values for other packing materials used in GPC.