Scaling AAV9 Manufacturing: A Blueprint for Clinical-Grade Gene Therapy Production

Learn how to overcome AAV9 bottlenecks with an open-source, high-yield manufacturing blueprint designed to accelerate clinical-grade gene therapy production.

Scalable AAV manufacturing remains one of the most persistent bottlenecks in advancing gene therapies to the clinic. Achieving high viral titers through triple transfection is notoriously difficult, and purifying drug substance to acceptable quality using scalable techniques like liquid chromatography has challenged the field for years. For many emerging CGT programs, access to a robust, license-free bioprocess only adds to those barriers.

Landmark Bio, an advanced cell and gene therapy CDMO focused on bridging the gap from bench to patient, undertook a project to address this directly. Supported by a grant from the National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL), Landmark Bio executed a pilot-scale AAV9 batch using the NIIMBL Viral Vector Program’s bioprocess framework — with the goal of creating an opensource platform that any organization can use to produce clinical-scale material. In confirming the process, the team optimized two critical unit operations: an upstream study using the AAViator+RevIT Transfection System that achieved greater than 10-fold improvement in viral titer, and a downstream effort to scale production and optimize chromatography for purity while maximizing yield. These findings offer a practical, transferable framework for further optimization — and a potential foundation for CGT programs that need a viable path forward.