Executive Summary
Effective AAV manufacturing functions as an interconnected system where upstream decisions directly influence downstream outcomes. Early, expert consultation on process design de-risks development by identifying and resolving potential scalability, purity, and yield issues long before they impact timelines. By integrating analytical strategy with process development from the outset, programs can build a robust, phase-appropriate CMC package that accelerates the path to IND.
The ratio of full to empty AAV capsids originates from upstream process parameters established during initial cell culture and transfection, directly impacting downstream purification. A manufacturing process designed for scalability anticipates these interdependencies, ensuring that decisions made at the 2L scale are viable at the 500L scale. This requires a deep understanding of vector biology and the physics of purification.
Our approach begins with a comprehensive evaluation of the entire manufacturing train, from plasmid design and cell line selection to the final formulation. We focus on key upstream drivers of yield and quality:
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Plasmid & Transfection: Optimizing plasmid ratios and transfection reagents to maximize vector expression while minimizing cellular toxicity.
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Cell Culture Platform: Selecting between adherent and suspension systems based on the specific AAV serotype, intended scale, and program timeline.
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Harvesting Strategy: Defining the optimal timepoint and method for harvest to maximize recovery of viable vector particles.
The success of the downstream purification cascade is predicated on the quality of the material from the upstream process. Our consultation focuses on designing efficient, scalable purification trains that consistently deliver a high-purity product. This includes chromatography resin selection, buffer optimization, and filtration strategies designed to separate full capsids from empty capsids and other impurities. This level of detailed planning provides a stable foundation for development. As one biotech partner noted, “The key people from UPenn Vector Core joined Franklin Biolabs and our partnership transitioned without interruption… Franklin Biolabs is integral to our AAV-vector based gene therapy candidate development.”
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Analytical development must run in parallel with process development. Complex therapeutic strategies, such as dual-vector systems for gene editing applications, introduce significant manufacturing challenges related to ensuring consistent product ratios and co-transduction efficiency (PMID: 26829317). A robust analytical platform is required to characterize these complex products and guide process improvements. By defining quality attributes early and developing assays to measure them, we create a data-driven feedback loop that accelerates optimization.
This integrated approach, combining upstream and downstream expertise with a forward-looking analytical strategy, is executed within our >100,000 sq ft facility. This methodology has supported our team’s 100% IND approval success rate since 2019. With the launch of Franklin Biolabs in 2024, this same experienced team provides our partners a clear, predictable path from discovery to clinical readiness, typically within an 18-24 month IND timeline.
