Optimizing Capture And Polishing Steps In An rAAV Purification Process

Scalable, cost-efficient, and robust filtration and chromatography-based processes are required for the purification of adeno-associated virus (AAV) in bioprocessing.

The key factors for an effective AAV purification process are high overall yields and maximized empty capsid product impurity reduction. After harvest by detergent cell lysis and DNA degradation by nuclease treatment, the feed is clarified followed by a concentration and buffer exchange step that reduces the volume to prepare for the capture step. Affinity capture is an efficient way to specifically bind and concentrate the target and at the same time achieve a high impurity removal.

We adapted an affinity capture step for rAAV2 and rAAV5, followed by a polishing step with rAAV5 to improve the yield of rAAV full capsids.

In this article, we show how we developed an efficient purification process for recombinant AAV (rAAV). The chromatography purification described includes affinity capture to maximize recovery with efficient impurity removal.