Preparation of Mix-mode Chromatographic Supports Based on Gigaporous Polymer Microspheres

The mix-mode chromatography supports were prepared, including anion exchange and hydrophobic interaction chromatography. This matrix was based on the gigaporous polymer microspheres from copolymer of glycidyl methacrylate and ethylene glycol dimethacrylate. The microspheres were modified by poly (ethylene imine) and butyl glycidyl ether for mix. mode chromatography. The effects of ion capacity and hydrophobic ligand density on protein capacity and recovery were evaluated. The result indicated that the protein capacity and recovery increased with ion capacity of the supports in the range of 0.2-0.5 mmol/mL. The max capacity was 40 mg/mL and the protein recovery could reach more than 90%. The hydrophobic interaction chromatography occurred when hydrophobic ligand density was more than 0. 03 mmol/mL. The back pressure on these supports was less than 2 MPa at 2000 cm/h. In the same time, the high resolution could be retained even at high flow rate of 2880 cm/h for purification of IgG from human serum. Therefore, this support showed a large potential in high through. put separation.