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

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 (2.5-5 cm diameter) are not usable due to irregular polymerization phenomena. Preparing thin disks of 0.5 cm height with similar diameter proved to be an excellent alternative. Disks could be stacked and run in a chromatographic setup. 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 sevenfold, we observed a 12-fold increase of ligand density and a sevenfold 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.