Clarification of recombinant adeno-associated virus (rAAV) & lentivirus from adherent culture

Abstract

In recent years the cell and gene therapy industries have been rapidly expanding, with two of the most utilized viral vector classes being adeno-associated virus (AAV) and lentivirus. With clinical success comes the need to develop and scale-up efficient manufacturing processes. As both of these vectors are produced in cells, the first step in their purification is to clarify them from the cell culture. There are many technologies traditionally used for cell culture clarification but given the projected manufacturing scales and need for single-use con-sumables a combination of depth and membrane filtration is a logical fit for batch processing of viral vectors. This work focuses on developing filtration-based clarification processes for both AAV and lentivirus. The data shows robust turbidity reduction and step yields across batches, scales, and AAV serotypes. We discuss how capacity can be impacted by feed-stream characteristics and how capacities translate to manufacturing footprints. Finally, we discuss some process considerations that are unique to viral vector processing and critical to successful vector harvest.