By Thomas Parker, Youness Cherradi, Ph.D., Claire Scanlan, And Elina Gousseinov, MilliporeSigma
As with recombinant proteins and antibodies, recombinant adeno-associated viruses (rAAV) are manufactured using cell culture processes, typically using insect (Sf9) or mammalian (HEK293) cells.
During early clinical phases or for low-dosage applications, rAAV production tends to be performed using adherent cell cultures; however, for larger late-stage clinical- and commercial-scale processes (>200 L), suspension cell culture in bioreactors is preferred.
After transfection or infection to produce rAAV, lysis is typically performed to release the virus from the cells. While mechanical or salt-based lysis is possible, detergent lysis is often accomplished using a surfactant, such as Triton X-100 or Tween® 20. Breaking open the cells releases large quantities of DNA into the bioprocess fluid, which generally is degraded with the addition of a nuclease such as Benzonase® endonuclease.
Normal flow filtration (NFF) using depth and membrane filters is the most popular approach for clarification of rAAV, as NFF generally provides good yields for rAAV vectors at a reasonable cost with minimal process development complexity. Depth filters can clarify a wide range of cell culture feed streams in a single-use format. There are several types of depth filters available. Learn more about options for depth filters and what is best for your application.