Related Experiment Video
Updated: Aug 5, 2026
08:17
Production of High-Yield Adeno Associated Vector Batches Using HEK293 Suspension Cells
Published on: April 26, 2024
Backbone-Minimised Nanoplasmid DNA Systems Enable High-Titre AAV Production in Suspension HEK293 Platforms
Lewis Hall1,2, Michael J Fiore2, Joel Abramovich2,3
1Department of Biochemical Engineering, University College London, London WC1E 6BT, UK.
Pharmaceutics
|July 28, 2026
Summary
Using compact Nanoplasmids™ DNA significantly boosts adeno-associated virus (AAV) production by up to tenfold. This scalable gene therapy manufacturing approach reduces costs and improves safety by minimizing bacterial plasmid DNA.
Area of Science:
- Gene Therapy Manufacturing
- Bioprocessing Technology
- Vector Biology
Background:
- Scalability and cost are key limitations for adeno-associated virus (AAV) gene therapy manufacturing.
- Current AAV production relies on triple-plasmid transfection using large bacterial plasmids, impacting cost-of-goods and productivity.
- Limited innovation exists in upstream AAV production processes compared to capsid engineering.
Purpose of the Study:
- To evaluate a compact Nanoplasmid™ DNA system for enhancing AAV upstream productivity.
- To compare Nanoplasmid™ DNA systems against conventional pUC-based plasmids in HEK293 suspension platforms.
- To assess the impact of full-system backbone minimization on AAV yield.
Main Methods:
- Utilized Nanoplasmid™ DNA and conventional pUC-based triple-plasmid systems in HEK293 suspension cells.
- Employed response surface Design of Experiments for platform optimization.
- Constructed and evaluated hybrid plasmid configurations to determine component-level contributions.
Main Results:
- Complete replacement with Nanoplasmid™ DNA yielded up to a 10-fold increase in vector genome titre compared to conventional plasmids.
- Hybrid systems did not achieve similar yield improvements, indicating the necessity of full backbone minimization.
- Productivity gains were consistent across different transfection reagents and suspension media.
Conclusions:
- Nanoplasmid™ DNA offers a scalable, capsid-independent method for upstream intensification in AAV production.
- This approach improves yield without affecting capsid biology, enhancing process robustness and economic feasibility.
- Integrating Nanoplasmid™ DNA into manufacturing workflows can reduce bacterial plasmid elements, increasing safety and reducing costs for AAV therapeutics.
