Protein separation by high-performance membrane chromatography

High-performance membrane chromatography (HPMC) is a very effective Chromatographic method that combines the advantages of both membrane technology and column chromatography. In this work, proteins (myoglobin, conalbumin and soybean trypsin inhibitor) were separated by HPMC. The separation mechanism involved anionexchange, and the stationary phases were used anion CIM DEAE and QA disks (12x3 mm). Two types of mobile phase, buffer A (20 mM Tris-HCl, pH 7.4) and buffer B (buffer A+1 M NaCl) were used. As the amount of sodium chloride dissolved in buffer linearly increased, the retention time shortened and the resolution of the components was greatly improved. The optimum mobile phases and operating conditions were experimentally determined. From the experimental results, the proteins were separated within 2 min at a mobile phase flow rate of 4 ml/min.