Rapid Purification of Molecular Chaperonins by Flowthrough Chromatography with Customized Biporous Anion-Exchanger

Summary

A rigid spherical biporous matrix was prepared by radical suspension polymerization. Functionalized with diethylamine, a biporous ion-exchange resin (IER-B) was obtained. The properties of IER-B were compared with the microporous ion exchange resin (IER-M) in pore size distributions and specific surface areas. Batch adsorption showed that IER-B and IER-M had comparable static adsorption capacities for BSA, i.e., 55.6 and 63.8 mg · g^-1 wet resin, respectively. The dynamic capacity of IER-B column decreased slightly with increasing the flow rate, which maintained at 38 mg · mL^-1 bed at a flow rate of 30 cm · min^-1. In contrast, the dynamic capacity of IER-M column decreased drastically to a value of only 13 mg · mL^-1 bed at a flow rate of 24 cm · min^-1. Then, the IER-B column was used for the purification of molecular chaperonins GroEL and GroES. At elevated flow rates from 2.55 to 10.1 cm · min^-1, the resolution and capacity of the column for GroEL and GroES purification was not affected by increasing flow rate. The results indicate that the biporous resin was promising for high-speed chromatographic purification of proteins.