AAV Production Using Microcarriers

In the industrialization of gene therapy, large-scale production of viruses has become a limiting factor, as gene therapy projects enter clinical phase III or get approval into the market.

The most widely used systems to produce adeno-associated virus (AAV) are static systems and other viruses using HEK293 cell transfection.

The typical and convenient way for industrialization is suspension culture, but the quality and yield of the target products will be affected by the differences in culture process between adherent cells and suspended cells. Thus, establishing a large-scale HEK293T cell culture process for virus production has become increasingly important.

Microcarriers are 60 to 250 micron support matrices allowing for the growth of adherent cells in a suspension system and can be made from various materials such as glucan, glass, and other synthetic polymers. Microcarrier culture combines the advantages of adherent and suspension culture and enables the suspension culture of cells in adherent state. It can produce large quantities of cells without the need for large-capacity equipment.

Obio Technology scientists have successfully adapted an adherent HEK293 cell line to grow in shaker flasks in animal component-free and serum-free suspension conditions. The scientists compared adherent, suspension, and microcarrier culture for AAV production and now believe microcarrier culture is suitable for large-scale production of viral vectors to support future gene therapy manufacturing processes.