LAUSR

Background & Aim The approval of gene therapies delivered by Adeno-Associated Viruses (AAVs) has resulted in an increased commitment to adopt AAV as the vector of choice to address a broad range of diseases. The increased demand for AAV is a challenge, which will probably only be fulfilled by adoption of platform processes, similar to those developed for monoclonal antibodies. An affinity capture step can be applied to most AAV serotypes and appears to be part of a nascent platform purification process. However, a post affinity step, most often ion-exchange chromatography, is required to remove the remaining host cell proteins and DNA, and to separate full capsids, containing DNA payload, from empty capsids. A robust process is difficult to develop for these chromatography steps and as dilutions must be performed after the affinity step, volumes to load can be large compared to the titer. Methods, Results & Conclusion To address these challenges, we have investigated the performance of membrane chromatography as a polishing step for purification of AAV serotypes 2, 5 and 9. We compare the performance for contaminant removal compared to commonly used resin based approaches to show the potential of membrane chromatography as a platform-able polishing step for AAV purification. Membrane chromatography can be loaded 40 times more quickly than conventional chromatography resins, which can lead to much higher productivities and greatly speed the purification step.