Transient expression systems have proven useful for generating significant amounts of material in short periods of time, which can be particularly valuable in the early stages of development when stable cell lines are unavailable. However, in the case of most virus or virus like particle production, the product of interest is toxic to the host cells and poses various problems for generating stable cell lines. Thus, to alleviate development time and complications, transient transfection processes can serve as a reasonable alternative.<br/><br/>In our work, we focus on gaining a deeper understanding of transient expression of VLPs to achieve enhanced predictability and process improvements. Our transient model focuses on the analysis of chikungunya virus like particle (VLP) expression using chemical (polyethylenimine) and physical (electroporation) based transfection methods with suspension HEK 293 cells. We use flow cytometry to visualize and interpret plasmid delivery and the resulting cellular responses. Additionally, we utilize the AMBR 15 microbioreactor system to screen for optimal transfection/expression conditions for enhanced productivity. Ultimately, our work offers significant insight on the kinetics of transient transfection/expression and the importance of the environmental conditions for successful VLP production.

Investigating and understanding process improvements for transient production of virus like particles in HEK 293 cells

253rd National Meeting (2017)

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