Process Optimization For Lentivirus Production In The iCELLis Nano Using A Chemically Defined, Animal Component Free HEK293 Media

By Sofia Pezoa, Ph.D., Director of Cell Culture, InVitria

Use of FBS in large scale clinical manufacturing applications represents a significant risk due to potential adventitious agent contamination, unreliable supply chain from limited global supply, and considerable variability in vector yield due to lot-to-lot composition variations. To circumvent the many challenges and drawbacks associated with using serum in viral vector production media for adherent cells, we utilized recombinant human proteins to formulate a chemically defined, serum-free and animal component free media that is optimized specifically for production of viral vectors in 293 and 293T cells in the iCELLis fixed bed bioreactor, called OptiPEAK HEK293t®. We therefore performed optimization studies in OptiPEAK HEK293t to determine the optimal parameters for LV production in the iCELLis Nano bioreactor. The parameters tested included cell density at the time of transfection, linear speed post transfection, pH as well as Dissolved Oxygen (DO) post transfection. Our results demonstrate that high cell densities at the time of transfection were negatively correlated with virus titer. We also observed that a high linear feed rate can be used post transfection without compromising virus titer. Taken together, we have demonstrated that the iCELLis Nano Bioreactor can be used with ACF conditions to achieve high lentivirus titer for clinical manufacturing.