Solvent Modulation of Column Chromatography

A majority of column chromatographies use only selected salts, e.g., ammonium sulfate, NaCl, Citrate and phosphate in hydrophobic interaction chromatography (HIC) and NaCl in ion exchange and dye affinity chromatographies. Alternatively, a pH range below or above the neutral value is often used to reduce affinity interactions, e.g., in Protein-A or dye affinity column chromatography. Although these parameters are easily manipulated, they are not necessarily the optimal conditions for high recovery and resolution of the proteins. So-called co-solvents have been used, although to a limited extent, to manipulate performance of column chromatography. Here the term co-solvent is used to indicate its relatively high concentrations required for these applications, meaning that it also serves as solvent along with water. Ethylene glycol and MgCl2 have been used to elute specific antibodies from antigen-affinity column. Arginine has also been used for the same purpose. Arginine has much wider applications for various column chromatographies, including size exclusion chromatography (SEC), HIC and affinity chromatography. Polyethylene glycol and glycine have also been used to improve the performance of HIC and hydroxyapatite chromatography. This review summarizes these applications of co-solvents for column chromatographies. Keywords: Solvent modulation, chromatography, affinity interaction, elution, co-solvent