AAV Production in HEK293 Cells
Type
Protocol
...other than PES. AAV particles stick to many other surfaces, but do not stick to PES. Using a PES filter will...polyethersulfone (PES) filter system, Nalgene, 565-0010 (or for larger volumes 1000 mL 0.45 µm Rapid-Flow PES Filtration...supernatant as follows: Filter through a 0.45 μm PES membrane. Add 40% PEG such that the final PEG concentration...sodium bicarbonate, Corning 25-035-CI (optional) 1 M HEPES, HyClone SH30237.01 (optional) L-alanyl-L-glutamine..., 7.5 mL of 7.5% Sodium Bicarbonate, 7.5 mL 1 M HEPES to 750 mL DMEM + 1 g/L glucose. 1 mg/mL polyethylenimine...Often AAV2 are harvested at 48–72 h, while other serotypes are harvested at 96–120 h. Optional: perform a...