Use of sulfated cellulose membrane adsorbers for chromatographic purification of cell cultured-derived influenza A and B viruses

摘要

Influenza virus particles can be purified by pseudo-affinity chromatography using sulfated carbohydrate matrices. In this study, we compared the binding capacity and the purification performance of two bead-based resins and one membrane adsorber for three influenza virus strains (H1N1, H3N2 and B) produced in MDCK suspension cells in a chemically defined medium. The dynamic binding capacity for the sulfated cellulose membrane adsorbers was consistently higher than for the resins (8 to 22-fold). Confocal microscopy demonstrated, that while virus particles uniformly covered the complete surface of the membrane, the virus particles only bound to the external surface of the sulfated beads. Overall, recovery of virus varied between 66% and 81%. Total protein and DNA removal were > 74% and > 96%, respectively. Statistically significant differences in the purification performance were more often observed between strains than between matrices. In particular, H3N2 influenza virus particles could only be purified using the sulfated membrane adsorbers. Due to the higher operating flow rate and binding capacity, the productivity with the membrane adsorbers was on average 25-times higher than with the resins.