Successful Capture DoE Studies For A Range Of AAV Serotypes To Reduce Manufacturing Costs, Accelerate Development

By M. Langhauser, R. Staffler, J. Trommer, C. Mantzoros, M. Boscher, B. Larena Carino, A. Youssef, and K. Heller

Adeno-associated virus (AAV) is the most widely utilized vector in in vivo gene therapy due to its versatility and effectiveness. Achieving optimal vector potency in specific target cells or organs while minimizing off-target effects and immunological challenges relies on selecting from numerous naturally occurring AAV serotypes and engineered capsids. Over the past two decades, capsid engineering technologies have significantly advanced, enabling further customization of AAV vectors for desired applications. However, capsid choice influences both upstream and downstream manufacturing, requiring tailored assay development and process optimizations.

Key upstream considerations include starting material design and transfection optimization. For downstream processing, capsid-specific adaptations are essential at every stage, particularly during the affinity chromatography capture step. Using a design of experiment (DoE) approach, this study identified optimal elution pH and conductivity conditions for AAV2, AAV5, and AAV8, revealing unique behavior for AAV5. Compatibility with subsequent full/empty particle separation was also confirmed.

The findings underscore the importance of selecting affinity resins with a wide dynamic range and high affinity, enabling robust, cost-effective manufacturing of high-quality AAV vectors. Download the poster to explore detailed data and insights for optimizing AAV manufacturing processes!