F. Bollmann, M. Noverraz, G. Pressac
Abstract
This study explores monolithic chromatography as an alternative to conventional resin- and ultracentrifugation-based methods for purifying adeno-associated viruses (AAVs). Leveraging monoliths’ convective mass transport for high resolution and capacity, lower pressure drops, higher flow rates, and reusability, we implemented a two-step approach for AAV8 purification, using CIMmultus® SO3 for initial capture and trialing three different chemistries (CIMmultus® QA, CIMmultus PrimaS®, and CIMmultus® PrimaT) for polishing. A design of experiments approach optimized the capture process, while the polishing steps effectively separated empty and full capsids, achieving high recovery and impurity clearance, including empty capsids. The scalability of the process was demonstrated, showcasing the robustness and efficiency of monolithic chromatography as a scalable alternative to conventional AAV purification.
