High Performance Anion-Exchange Chromatography of Proteins

Abstract

High performance anion-exchange chromatography using an aqueous solvent system is presented for the analysis and preparation of proteins. Ten purified proteins, having a molecular weight of 10,000 - 190,000 daltons and an isoelectric point of 3.9 - 8.5, were applied to a diethylaminoethyl (DEAE) polymer-based column and eluted within 60 min by a linear salt gradient of NaCl in 0.05 M Tris-HCl buffer at pH 7.5. The retention time of protein increases linearly with a decreasing order of the pI value of the protein in this system. By the application of this method, neuron-specific enolase and ceruloplasmin were purified from partially-purified preparations of these proteins respectively, and a series of isoforms of brain S100 protein were separated from each other. This column is capable of separating proteins in high speed, high resolution, large capacity, and in considerably high recovery of proteins without losing the biological activity.