Molecular-Sieve Chromatography of Proteins on Granulated Polyacrylamide Gels

Abstract

Abstract A method for molecular-sieve chromatography of proteins on columns of granulated polyacrylamide gels is described in detail. Distribution coefficients (K D and K av) are given for the model proteins γ globulin, bovine serum albumin, ovalbumin, ovomucoid, pepsin, myoglobin, and cytochrome c, as well as certain other proteins, between 0.5 M NaCl and polyacrylamide gels of 30 different compositions. The results are interpreted in terms of the Ogston-Laurent-Killander theory of molecular-sieve chromatography, and empirical equations are derived relating the fundamental parameters L and r of this theory to the gel composition. The results are discussed in terms of the gel structure and a method for estimating the average effective pore radius in polyacrylamide gels of different compositions is derived and compared with experimental findings.