Scalable lentiviral vector production using stable producer cell lines in perfusion mode
September 17-21, 2017
Lentiviral vectors (LVs) are becoming an important tool in gene and cell therapy and are being utilized in several clinical studies for rare and more frequent genetic and acquired diseases, as well as in cancer therapies. However, two major challenges need to be overcome in order to generate enough material to treat patients: First, current production platforms result in low titers (stable producer cell lines from adherent cell lines) or are not amenable to large scale production (LV produced by transfection). Next, LVs are known to have a low temperature stability. To address these two challenges, the National Research Council Canada has developed packaging cell lines and stable producer cell lines for the production of LVs which can grow in suspension in serum-free media and produce LV in the 106 TU/ml range without optimization. Furthermore, productions are performed in perfusion mode in order to operate at high cell densities and address the low LV stability. To facilitate titration, a producer cell line for LV expressing GFP regulated by the strong constitutive CMV promoter was generated (HEK293SF-LVP-CMVGFPq-92). Transcription of Rev and the envelope protein (VSVG) is under the control of the tetracycline and cumate switches, which means that addition of doxycycline and cumate is required to induce the production of LV. Results obtained demonstrate that the system is scalable (Fig. 1A), and up to 15 fold increase in total yield was obtained in perfusion mode when compared to batch mode (Fig. 1B), using perfusion rates of 0.65 to 1 vessel volume exchange per day after induction.
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Aziza Manceur, Stéphane Lanthier, Sonia Tremblay, July Dorion-Thibaudeau, Julia Transfiguracion, Hafida Aomari, Rénald Gilbert, and Sven Ansorge, "Scalable lentiviral vector production using stable producer cell lines in perfusion mode" in "Integrated Continuous Biomanufacturing III", Suzanne Farid, University College London, United Kingdom Chetan Goudar, Amgen, USA Paula Alves, IBET, Portugal Veena Warikoo, Axcella Health, Inc., USA Eds, ECI Symposium Series, (2017). https://dc.engconfintl.org/biomanufact_iii/77