top of page


Commercial Scale Production (1000L) of AAV Using the Gibco™ CTS™ AAV-MAX Production System

J Zmuda(1) C Y Liu(1) K Thompson(1) C Rasmussen(1) P Decaria(1) E Jackson-Holmes(1) I Pringle(1)

1:Thermo Fisher Scientific

Until relatively recently, AAV production has taken place primarily in adherent cultures using 293T cells in the presence of fetal bovine serum (FBS).  Adherent systems suffer from a number of shortcomings including difficulty in scaling up, the presence of the SV40 large T antigen in the producer 293T cell line as well as cost, consistency and regulatory considerations from the use of animal sera. To address these shortcomings, we developed the Gibco™ CTS™ AAV-MAX Helper-Free AAV Production System (AAV-MAX), an all-in-one, chemically defined, suspension-based AAV production system that allows for scalable, high titer production of AAV viral vectors in a non-293T cell lineage.  The system comprises all of the components required for scalable AAV production: a clonally-documented, 293F-derived producer cell line (VPC 2.0), a chemically-defined growth and expression medium, a production enhancer, a cationic lipid-based transfection reagent and booster, a chemically-defined complexation buffer, and a polysorbate 20-based lysis buffer. 

In the present study, we demonstrate consistent AAV production and cellular kinetics of expression from ml scale in multi-well plates and shake flasks to 50-1000L scales in single-use stirred-tank bioreactors using AAV-MAX.  Additionally, we provide guidance around scalable downstream purification and strategies for analytical characterization of the AAV particles for a seamless end-to-end AAV production process.  These data demonstrate that AAV-MAX is a flexible platform for scalable, high-titer AAV production in a streamlined protocol that maximizes viral titers while reducing time and cost of production, thus providing a suitable platform for AAV production from basic research through clinical and commercial production.

bottom of page