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Abstract:
In recent years, interest in macroporous matrices like membrane adsorber and monoliths for the purification of cell culture-derived viral particles and viral vectors increased steadily. This trend is mainly attributed to their hydrodynamic benefits compared to classical bead based resins, which allow to reduce the overall process time. Furthermore, in the case of membrane adsorbern the availability of prepacked disposable capsules eliminate the need for packing, cleaning, sanitization and their validation.
Here we give an overview on downstream processing of cell culture-derived influenza and vaccinia virus. Influenza viruses (A/Wisconsin/67/2005, A/Puerto Rico/8/34, B/Malaysia/2506/2004) were derived from Madin-Darby canine kidney (MDCK) cell cultures, and the Modified Vaccinia Ankara virus (MVA-BN®) from primary chicken embryo fibroblasts (CEF). Specific focus will be the use of affinity membrane adsorption (MA) chromatography as capturing step. In addition, purification based on pseudo-affinity MA chromatography is compared to bead-based resins (Cellufine® sulfate) and conventional ion exchange MA.
For influenza and vaccinia virus particles a remarkable high degree of contaminant depletion is achieved by use of affinity MA compared to ion exchange MA. Up to 90% of the host cell DNA can be eliminated compared to the starting content. Contamination with host cell proteins can be reduced to less than 5% of the initial amount. Overall, product yields of up to 90% (influenza) and 80% (vaccinia) were obtained. Due to the increased volumetric throughput an up to 20-fold increase in productivity was achieved with MA compared to bead based resins.
