The success of AAV-based gene therapies relies on bioprocess technologies that can produce high-purity, cell-transducing capsids at clinically relevant scales. However, current expression systems generate a mix of full, partial, and empty capsids, whose proportions vary with serotype, transgene, and process conditions, posing safety risks such as immunogenicity and genotoxicity. Removing these impurities requires a complex anion-exchange chromatography (AEX) enrichment step, which must be tailored to specific AAV designs and sample characteristics. Developing effective AEX protocols is challenging due to numerous interdependent parameters, including stationary phase properties, flow rates, buffer compositions, and elution formats.
To address the challenges in AAV capsid purification, this project will leverage the combined expertise and resources of NIIMBL academic and industry partners, including advanced chromatographic technologies, modeling tools, and a diverse library of model AAVs. We will establish the Platform for Analytical Chromatographic Testing (PACT) to rapidly screen and validate AEX protocols across varied AAV designs, generating a comprehensive dataset of in-line, at-line, and off-line analytics. This data will inform the development of the Rulebook of AAV Analytical Chromatography (RAAC), a predictive framework linking chromatographic parameters to capsid purity outcomes. Together, PACT and RAAC will transform AEX method development from empirical trial-and-error to a systematic, data-driven approach, accelerating gene therapy manufacturing and supporting NIIMBL’s broader data initiatives.
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North Carolina State University
Bio-Rad Laboratories, Inc.
Bristol-Myers Squibb
Chromagenix
EMD Millipore Corporation
Joan & Sanford I. Weill Medical College of Cornell University
Refeyn Inc.
Rensselaer Polytechnic Institute
Sartorius Stedim
University of Florida
