Towards a scalable bioprocess for rAAV production using a HeLa stable cell line
Open Access
- 7 April 2023
- journal article
- research article
- Published by Wiley in Biotechnology and Bioengineering
- Vol. 120 (9) , 2578-2587
Abstract
The majority of recombinant adeno‐associated viruses (rAAV) approved for clinical use or in clinical trials areproduced by transient transfection using the HEK293 cell line. However, this platform has several manufacturing bottlenecks at commercial scales namely, low product quality (full to empty capsid ratio 3 × 1011 vg/mL and full to empty capsids ratio (>70%) at 2 L bioreactor scale. Moreover, the established downstream process, based on ion exchange and affinity‐based chromatography, efficiently eliminated process related impurities, including the Adenovirus 5 helper virus required for production with a log reduction value of 9. Overall, we developed a time‐efficient and robust rAAV bioprocess using a stable producer cell line achieving purified rAAV2 yields > 1 × 1011 vg/mL. This optimized platform may address manufacturing challenges for rAAV based medicines.Keywords
Funding Information
- Horizon 2020 Framework Programme
- Fundação para a Ciência e a Tecnologia
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