10.27.22 -- From Cells To Capsid Separation: A Scalable rAAV Process

The cell and gene therapy industry is striving to establish efficient large-scale production of viral vectors with current good manufacturing practice (cGMP). Review experiments for the adaptation of the HEK293 suspension cell line for transient transfection and scalable adeno-associated virus (AAV) production.

Explore how we expanded HEK293/HEK293T suspension cells from small cell culture (20 mL) in shake flasks up to a 10 L culture in a benchtop Xcellerex™ XDR-10 single-use bioreactor system. AAV was produced at high titers upon transient plasmid transfection in the bioreactor.

In this application note, we describe the performance of two assays for titer analysis of adeno-associated virus (AAV), serotype 2 and serotype 5, based on surface plasmon resonance (SPR). Both assays provide repeatable results and correlate well with established total AAV capsid titer assays using ELISA.

Scalable, cost-efficient, and robust filtration and chromatography-based processes are required for the purification of adeno-associated virus (AAV) in bioprocessing. In this article, we show how we developed an efficient purification process for recombinant AAV (rAAV).

Optimizing the polishing step of your AAV process for the separation of full capsids from empty capsids for each serotype is essential. We show how Capto™ Q resin with dextran surface extenders, magnesium chloride (MgCl₂), and the elution salt type (especially for rAAV9) significantly enhance separation.