Preparation and characterization of large-format macroporous cryogel disks for use in affinity chromatography and biotechnological applications

Abstract

We have prepared and evaluated larger format phage-bound epoxy-cryogel columns in order to increase the yield of bound target. Freezing thermograms showed that larger column formats (2,5 cm-5 cm diameter) are not usable due to irregular polymerization phenomena. Preparing thin discs of 0,5 cm height with similar diameter proved to be an excellent alternative. Discs could be stacked and run in a chromatographic set up. In this way we could increase the matrix volume, ligand binding capacity and finally the yield of bound target. By increasing the column volume about 7 fold, we observed a twelve fold increase of ligand density and a 7 fold increase in the yield of protein recovery in a column where phages were attached without spacer and a 10 to 34 fold increase in a spacer column, depending on the spacer used.