Building A Chromatography Platform For Purification And Full Capsid Enrichment On AAV8

The development of gene therapy using Adeno-Associated Virus (AAV) vectors necessitates the efficient purification of full capsids to ensure therapeutic efficacy and minimize immunogenic risks associated with empty capsids. This study focuses on optimizing chromatography techniques for the purification and enrichment of AAV8 full capsids, specifically targeting improvements in anion exchange chromatography (AEX) following affinity chromatography.

AAV8 was produced in-house using suspension HEK293 cells and purified through a multi-step process involving depth filtration, tangential flow filtration, and affinity chromatography. The subsequent AEX development evaluated various resins, loading conditions, and buffers, employing dynamic light scattering (DLS), sedimentation velocity analytical ultracentrifugation (SV-AUC), and transmission electron microscopy (TEM) for analysis. Initial findings demonstrate a significant enhancement in full capsid percentage, achieving up to 88.7% enrichment, with the potential for further optimization.

This poster presents a comprehensive overview of the methodologies, experimental conditions, and outcomes, highlighting the efficacy of chromatography in the scalable purification of AAV vectors.