Abstract
A two step flow-through chromatography process is proposed as an universal approach to purify viruses. A resin column with reduced surface area was developed for the first step to remove bulk of the host cell protein (HCP) from a viral feed stream while allowing most of the virus to flow-through. For the second step a chromatographic separations strategy using a primary amine membrane adsorber and multivalent ions in the mobile phase was developed. This enabled selective binding of host cell DNA (hcDNA) to the membrane and complete recovery of virus in the flow-through mode. The techniques were evaluated using cell culture grown influenza virus and bacteriophage feed streams. Virus recoveries of >70-80% and 100% were achieved for the column and membrane approaches respectively. The column cleared > 80% of the HCP and the membrane adsorber reduced whole hcDNA levels to <10 ng.
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