Improved Lentiviral Vector Biomanufacturing for Cell and Gene Therapy Applications

The aim of this NIIMBL-funded project is to provide the industry with a lentivirus production process that yields large quantities of highly potent vector with fast facility turnaround times.

Project status

100% Completed

Industry Need

Current production processes, in which host mammalian cells are used to generate

lentivirus, are inefficient, resulting in low yields, lack of scalability, and high biomanufacturing costs.

With clinical trials and FDA approval of T-cell immunotherapy, efficient lentiviral production processes

appropriate for large scale manufacturing are needed.

Solution

The aim of the NIIMBL-funded project was to provide the industry with a lentivirus production process that yields large quantities of highly potent vector with fast facility turnaround times.

Outputs/Deliverables

Establish a microscale screening platform that allows for high density growth of HEK293 suspension cells.
Demonstrated high density perfusion culture of HEK293 suspension cells in bioreactor format.
Identify optimal LVV production conditions in HEK293 suspension cells in commercially available medium that yield high titer lentivirus.
Identify mobile phase conditions that enhance lentivirus stability.
Establish robust, fast and scalable chromatography capture step for purification of LVV.
Establish novel, robust and scalable flow through chromatography purification step.
Established a suite of robust precise and accurate analytical methods for Lentiviral Vector manufacture.

Impacts

Improved viral vector production process yield, processing times, and measurement accuracy

Development of a platform process to mitigate lentiviral vector supply shortages for cell and gene therapy

Publications

Ghosh, R., Koley, S., Gopal, S., Rodrigues, A. L., Dordick, J. S., & Cramer, S. M. (2022). Evaluation of lentiviral vector stability and development of ion exchange purification processes. Biotechnology Progress, 38(6). https://doi.org/10.1002/btpr.3286