Lentiviral vectors (LVVs) are emerging as a promising tool for gene and cell therapy. Due to the ability of LVVs to deliver genetic payloads to both dividing and non-dividing cells, they possess significant potential for introducing therapeutic genes to cells of interest. LVVs are typically produced by transfecting envelope, packaging and transfer plasmids into Human Embryonic Kidney 293 (HEK293) or other suitable cell lines. Nevertheless, several challenges remain in the production process including low titers and a lack of suitable scale-up production. To address these concerns, we developed a high-throughput microwell platform for upstream transfection optimization leading to higher yields of LVVs. Several commercially available cell media were evaluated for their ability to support high cell densities. BalanCD – a commercially available HEK293 suspension cell medium was able to support both high cell densities and generate high transfection efficiencies, and was, therefore, selected for further screens. To optimize transfection, screens were performed to establish optimal amount of DNA/cell, viable cell density at time of transfection, optimal ratios among packaging, envelope and transfer plasmids, and the ideal transfection reagent to DNA ratio. Comparisons of optimal conditions across scales were then made for both batch and perfusion scale-up modes.<br/>

Scaled-down, high-throughput optimization of transfection for lentivirus production for use as gene therapy vectors

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.