Harvesting Adeno-Associated Virus from Transfected Mammalian Cells

Begin with a multi-layered cell chamber containing transfected mammalian cells producing adeno-associated virus vectors. The cell chamber helps generate a high-yield viral harvest.

Shake the chamber vigorously to dislodge the adherent cells, releasing virus-containing cells and extracellular viral particles into the medium.

Transfer the contents to a tube and centrifuge to pellet the virus-containing cells.

Collect the supernatant containing the extracellular virus and transfer it into a storage bottle.

Resuspend the pellet in lysis buffer and incubate.

Enzymes and detergents in the buffer lyse the cells and release the intact intracellular viruses into the solution.

Centrifuge the lysate to pellet the cellular debris. Collect the supernatant containing the virus and add it to the same storage bottle containing the extracellular virus.

Store the crude viral suspension at low temperature until further purification.

After incubation, shake the cell culture chamber vigorously to dislodge the cells until the media appears cloudy, and pour it into four 500-milliliter centrifuge tubes.

Pellet the cells by centrifugation at 18,000 x g for 30 minutes at four degrees Celsius. Pour the clarified supe