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Lentiviral Vector Production from a Stable Packaging Cell Line Using a Packed Bed Bioreactor.
Alicia D Powers1, Jason E Drury1, Christopher F Hoehamer2
1Department of Therapeutics Production & Quality, St. Jude Children's Research Hospital, Memphis, TN 38105-3678, USA.
Molecular Therapy. Methods & Clinical Development
|September 30, 2020
Summary
Scalable production of lentiviral vectors (LVVs) for gene therapy was achieved using the iCELLis bioreactor. This method offers high viral yields comparable to traditional cell factories, ensuring efficient cellular gene therapy applications.
Area of Science:
- Biotechnology and Bioprocessing
- Gene Therapy Vector Production
- Cellular Engineering
Background:
- Self-inactivating lentiviral vectors (LVVs) are crucial for genetic modification in cellular gene therapy, particularly for T cells and hematopoietic stem cells.
- Increasing demand for LVVs necessitates scalable and controllable production methods beyond traditional suspension or adherent cultures.
- Existing methods often rely on HEK293T cells in suspension bioreactors or serum-containing adherent cultures.
Purpose of the Study:
- To optimize and scale up the production of a specific LVV (GPRTG-EF1α-hγc-OPT) using the iCELLis Nano bioreactor system.
- To compare the productivity and efficacy of LVVs produced in the iCELLis Nano with those manufactured under current Good Manufacturing Practice (cGMP) using cell factories.
- To evaluate the suitability of the iCELLis Nano for producing LVVs for treating X-linked severe combined immunodeficiency.
Main Methods:
- Production of GPRTG-EF1α-hγc-OPT LVV using a stable packaging cell line in iCELLis Nano bioreactors (0.53 and 2.6 m² surface area).
- Optimization of key parameters including fetal bovine serum (FBS) concentration, post-induction pH, and induction day.
- Comparison of viral yields and transduction efficiency of iCELLis-produced LVVs against those from cGMP cell factories using purified CD34+ cells.
Main Results:
- Optimized production in the iCELLis Nano bioreactor achieved viral yields exceeding 2 × 10⁷ transducing units/mL.
- LVVs produced using the iCELLis Nano demonstrated transduction efficiency comparable to LVVs manufactured in cell factories.
- The study successfully scaled LVV production using the iCELLis Nano bioreactor system.
Conclusions:
- The iCELLis Nano bioreactor provides a scalable and efficient platform for high-yield LVV production.
- LVVs manufactured using this system maintain high efficacy, suitable for clinical applications like X-linked severe combined immunodeficiency treatment.
- This optimized bioprocess offers a viable alternative for meeting the growing demand for gene therapy vectors.
