Traditionally, VV- and MVA virus based vaccines have been grown in primary chicken embryo fibroblast cultures and purified either by sucrose cushion or sucrose gradient centrifugation, or by ultrafiltration. However, a potential shift from primary to continuous cell cultures would impose stricter requirements regarding the purity level of the vaccines, and a new generation of vaccine manufacturing processes is needed that include more sophisticated and innovative downstream techniques for purification.
Here, we report the development of an affinity chromatography of cell culture derived VV after an initial host cell homogenization and clearance centrifugation. The Vaccinia viral envelope protein A27L is known to bind to heparin.. Based on this, small scale chromatography experiments with heparinized polymer beads and cellufine sulfate beads in addition to heparinized cellulose membranes have been conducted. There we have found that membrane adsorbers are superior over bead based chromatography media in terms of efficiency and productivity.
Subsequent studies compared ion exchange membrane adsorbers with a heparinized membrane adsorber. The results indicate that the overall performance of the affinity chromatography in terms of virus capturing and contaminant removal is better than any of the tested ion exchange membrane adsorbers.
Hence, membrane affinity chromatography represents a valuable choice to capture VV particles in addition to the general advantages of membrane chromatography.