Scalable Production Of Recombinant AAV Vectors From A549-Based Packaging Cell Lines In The iCELLis™ System
Vector based on adeno-associated virus (AVV) are the subject of increasing interest for many gene transfer application including gene therapy. The current limitations of this technology is the lack of a versatile and scalable vector manufacturing process. In order to address this issue, we have developed A549-based stable packaging cell lines, which contain the AAV2 rep gene and the cap gene from various AAV serotypes. These cell lines efficiently assembler rAAV particles upon dual adenovirus infection (wild-type Ad5 followed by hybrid Ad/AAV), which is correlated with amplification and high-level expression of the integrated rep-cap genes. For production scale-up, we chose the high-cell density iCELLis™ system developed by ATMI. This technology consists of a disposable bioreactor system with a fixed-bed design operating in perfusion mode. The system is available in three is different scales, each with a range of available surface areas (0.5 – 5.3 m2, 500 – 1000 m2) depending on the chosen fixed bed volume and carrier compaction level, which are adjustable to suit the cell size, maximum cell density and required cell quantity. We present the result of a feasibility study performed with our A549-R2C8 cell line in the iCELLis nano fixed-bed bioreactor system.