Journal of Stem Cell Research & Therapy

Production of therapeutically relevant lentiviral vectors for clinical studies

7^th International Conference and Exhibition on Cell and Gene Therapy

March 15-16, 2018 | London, UK

Johann-Christoph Dettmann, Martin Meyer and Thomas Noll

Miltenyi Biotec GmbH, Germany

Posters & Accepted Abstracts: J Stem Cell Res Ther

Abstract:

The successful genetic engineering of patient-specific T cells with lentiviral vectors (LVV) expressing, e.g., chimeric antigen receptors for late phase clinical trials and beyond, requires the large-scale manufacturing of high-titer vector stocks. Their mass production remains a challenge and limits their potential therapeutic use, requiring robust, scalable and regulatorycompliant processes for industrial vector production. The state-of-the-art production of lentiviral vectors is based on 10 to 40 layer cell factories transiently transfected in the presence of serum. This manufacturing process is extremely limited by its labor intensity, open-system handling operations and its requirement for significant incubator space, plus costs and patient risk due to the presence of serum. To circumvent these limitations, this study aims to develop a stable and serum-free process to produce lentiviral vectors with PEI-mediated transfection. In addition, this study also focuses on the development of a production system not only using a GFP marker but also therapeutically relevant transgenes to overcome the limitations of GFP as process prediction model. Three different cell lines (HEK 293, HEK 293T, HEK 293FT) were investigated concerning their productivity of LVV and their growing behavior in the in-house serum free medium TransMACS. As part of this, a design of experiment was used to investigate the optimal conditions for PEI/DNA-transfection. Furthermore, this statistical approach was used to identify an ideal ratio between the four 3rd generation plasmids (transfer plasmid, envelope plasmid, packaging plasmids). In addition, several different transfection enhancers were investigated concerning their enhancing effects on productivity, comparing HEK cultures producing LVV encoding for different transfer genes.

Biography :

Johann-Christoph Dettmann completed his Master of Science in Biochemistry and Microbiology in 2014 at the University of Rostock, Germany. After this, he took up a Trainee position at the company, Miltenyi Biotec and is now working on his PhD thesis.
Email:johannd@miltenyibiotec.de