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Published on: June 19, 2013
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Scalable Lentiviral Vector Production Using Stable HEK293SF Producer Cell Lines
Aziza P Manceur1, Howard Kim2, Vanja Misic2
11 Human Health Therapeutics, National Research Council Canada , Montreal, Canada.
Human Gene Therapy Methods
|August 23, 2017
Summary
This study developed a scalable lentiviral vector (LV) production method using a stable HEK293 producer cell line. Perfusion strategies significantly increased LV titers, enabling large-scale manufacturing for gene and cell therapies.
Area of Science:
- Biotechnology
- Gene Therapy
- Cell Therapy
Background:
- Lentiviral vectors (LV) are crucial for gene and cell therapies.
- Large-scale production of LV remains a challenge for clinical applications.
- Suspension culture processes are needed for scalable LV manufacturing.
Purpose of the Study:
- To develop and optimize a scalable lentiviral vector production strategy.
- To improve lentiviral vector titers using a stable inducible producer cell line.
- To enable efficient large-scale manufacturing of lentiviral vectors.
Main Methods:
- Developed a stable inducible HEK293 producer cell line (clone 92) for LV production.
- Demonstrated scalability from shake flasks to bioreactors.
- Implemented perfusion strategies with acoustic cell filter technology for high-yield production.
Main Results:
- Achieved LV titers of 10^6 TU/mL without optimization.
- Improved titers to 3.2 x 10^7 TU/mL using perfusion strategies.
- Increased cumulative functional LV titers by up to 15-fold, reaching 8 x 10^10 TU/L.
Conclusions:
- The developed LV production strategy is scalable and amenable to commercial manufacturing.
- Perfusion-based production significantly enhances LV yield and efficiency.
- This method addresses the need for large-scale LV production for therapeutic applications.
