Optimizing and developing a scalable, chemically defined, animal component-free lentiviral vector production process...

Area of Science:

• Biotechnology
• Gene Therapy
• Cell Culture

Background:

• Lentiviral vectors (LVVs) are crucial for ex vivo gene-modified cell therapies.
• Current LVV production methods face challenges in scalability and cost, potentially limiting therapeutic applications.

Purpose of the Study:

• To develop and optimize a scalable, chemically defined, animal component-free LVV production process.
• To enhance functional LVV titers and facilitate cost-effective manufacturing.

Main Methods:

• Optimization of 2D static cultures, including cell seeding density, transfection timing, plasmid DNA concentration, and sodium butyrate addition.
• Transfer of the optimized process to an iCELLis Nano fixed-bed bioreactor operated in perfusion.
• Comparison of LVV titers using chemically defined medium versus conventional serum-supplemented medium.

Main Results:

• A 26.9-fold improvement in functional LVV titers was achieved through process optimization in 2D cultures.
• The optimized process in the iCELLis Nano bioreactor yielded titers of 1.2 × 10^6 TU/cm^2 in perfusion.
• Comparable functional LVV titers were obtained using a chemically defined medium and a conventional serum-supplemented medium.

Conclusions:

• A scalable, chemically defined, animal component-free LVV production process has been successfully developed.
• The optimized process significantly enhances LVV production efficiency and reduces associated costs.
• This advancement supports the broader application of LVVs in gene-modified cell therapies.