Improved Lentiviral Vector Biomanufacturing for Cell and Gene Therapy Applications

Performance Period: 1/1/2019 to 12/31/2020

Gene Therapy is an emerging area of therapeutics with the potential for curative treatment. Advances in genomics, gene editing methodologies, immunology and drug delivery technologies are driving a revolution in healthcare aimed at tackling the world’s most grievous illnesses by correcting genetic defects, enhancing cellular and tissue function, and improving production of cellular products or generating novel screening targets for drug discovery. A critical component of gene therapy is the development of a suitable viral vector that can be produced at large scale. Lentiviruses are a prominent viral vector under widespread consideration. Unfortunately, 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.

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. Specifically, upstream challenges will be addressed using high-throughput studies to optimize host cell growth and increase lentivirus production. These findings will be used to develop a cell culture process using transient transfection of high density HEK293 cells[1] adapted to serum-free medium. Downstream purification will be enhanced by advanced chromatographic separations and screening media reagents. These findings will be used to replace existing lentivirus purification processes with fast, high yielding flow through mode chromatography processes. In parallel, analytical technologies will be developed to enable manufacturers reduced feedback time for in-process samples as well as increased accuracy and precision of viral and infectious titer measurement. Ultimately, an integrated process including optimized bioreactor operations and downstream purification will be developed appropriate for scale-up. This NIIMBL academic-industrial collaboration will define and modify key factors with the goal of improving lentiviral production in a process appropriate for large scale biomanufacturing.