TY  - JOUR
U1  - Wissenschaftlicher Artikel
A1  - Hein, Marc D.
A1  - Kazenmaier, Daniel
A1  - van Heuvel, Yasemin
A1  - Dogra, Tanya
A1  - Cattaneo, Maurizio
A1  - Kupke, Sascha Y.
A1  - Stitz, Jörn
A1  - Genzel, Yvonne
A1  - Reichl, Udo
T1  - Production of Retroviral Vectors in Continuous High Cell Density Culture
JF  - Applied Microbiology and Biotechnology
N2  - Retroviral vectors derived from murine leukemia virus (MLV) are used in somatic gene therapy applications e.g. for genetic modification of hematopoietic stem cells. Recently, we reported on the establishment of a suspension viral packaging cell line (VPC) for the production of MLV vectors. Human embryonic kidney 293-F (HEK293-F) cells were genetically modified for this purpose using transposon vector technology. Here, we demonstrate the establishment of a continuous high cell density (HCD) process using this cell line. First, we compared different media regarding the maximum achievable viable cell concentration (VCC) in small scale. Next, we transferred this process to a stirred tank bioreactor before we applied intensification strategies. Specifically, we established a perfusion process using an alternating tangential flow filtration system. Here, VCCs up to 27.4E + 06 cells/mL and MLV vector titers up to 8.6E + 06 transducing units/mL were achieved. Finally, we established a continuous HCD process using a tubular membrane for cell retention and continuous viral vector harvesting. Here, the space-time yield was 18-fold higher compared to the respective batch cultivations. Overall, our results clearly demonstrate the feasibility of HCD cultivations for high yield production of viral vectors, especially when combined with continuous viral vector harvesting.
KW  - Gentherapie
KW  - Process Intensification
KW  - High Cell Density Cultivation
KW  - Perfusion Cultivation
KW  - Gene Therapy
KW  - Continuous Viral Vector Harvesting
KW  - Murine Leukemia Viral Vectors
Y1  - 2023
UN  - https://nbn-resolving.org/urn:nbn:de:hbz:832-epub4-27530
SN  - 0175-7598
SS  - 0175-7598
SN  - 1432-0614
SS  - 1432-0614
U6  - https://doi.org/10.1007/s00253-023-12689-9
DO  - https://doi.org/10.1007/s00253-023-12689-9
VL  - 107
IS  - 19
SP  - 5947
EP  - 5961
S1  - 15
PB  - Springer Berlin Heidelberg
ER  -