Figure 6.

Depiction of recombinant adeno-associated virus (rAAV) manufacturing technology. (1) Suspension HEK293 cells in the WAVE bioreactor 20/50EHT are transfected. (2) Approximately 48 hours post-transfection, cells are harvested into centrifuge bottles and pelleted at low-speed centrifugation. (3) Cell pellets are resuspended and then lysed using a sonicator. (4) Lysate is incubated with Benzonase for 45 minutes at 37 °C. (5) Lysate is clarified using low-speed centrifugation. (6) Clarified lysate is loaded onto iodixanol gradient and ultracentrifuged. (7) rAAV collected from the 40%/60% iodixanol interface are loaded onto AKTA FPLC and purified using anion exchange chromatography. (8) rAAV chromatography peak fractions are dialyzed into final formulation buffer. (9) Sterile filtration through 0.2 μm filter.