Generation Of A Universal AAV Packaging Cell Line For Use In Commercial Gene Therapy Manufacturing

By Marguerite Campbell, Kelly Geosits, Ngan Trinh, Khan Umaer, Ph.D., Srivanya Tummala, & Brian Tomkowicz, Ph.D., Viral Vector Research and Development, SK pharmteco Cell & Gene North America – King of Prussia, PA, United States

The growing demand for large quantities of high-quality viral vectors — produced at lower cost — is critical to advancing both rare disease therapies and broader clinical applications. To address this need, SK pharmteco’s Cell & Gene North America team has developed and validated a CGMP-ready, clonally derived, universal viral production cell line: SKPT-HEK293-4G9. This platform supports plasmid-based transient viral vector production from research to commercial scale.

Building on the serum-free, suspension-adapted SKPT-HEK293 parental line, we further engineered the SKPT-HEK293 AAV packaging cell line, incorporating stable integration of Rep, Capsid, and Ad Helper elements under a doxycycline-inducible promoter. With this design, simple transfection of a gene-of-interest (GOI) cis-plasmid enables single-step AAV production upon induction, eliminating the inefficiencies of multi-plasmid transient transfection. This approach significantly reduces process complexity, production costs, and batch variability.

We outline the workflow for generating stable pools and single-cell clones using the Cytena F.Sight cell printing platform. Our data demonstrate stable expression and production efficiency comparable to traditional triple-transfection methods.

Together with SK pharmteco’s established manufacturing, quality, and regulatory expertise, these off-the-shelf stable AAV capsid solutions provide a robust, scalable pathway to commercial manufacturing. By reducing the cost of goods (COGS), this platform has the potential to make gene therapies more affordable and accessible for patients with urgent, unmet needs.