16th International Adenovirus Meeting

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Poster presenter: Katja Vrabec
Title: Integrating Upstream and Downstream Processes for Enhanced Purity and Yield of Adenovirus 5

Poster abstract:

Optimizing adenovirus (AdV) production requires a comprehensive approach that addresses both the upstream and downstream processes, while ensuring process robustness. This presentation will focus on bioprocess development for improved AdV yield and purity, with simplification of operations in mind. We compare various industry-relevant upstream strategies for producing Ad5 and address each step of the purification process, while evaluating both adenovirus concentration and bioburden through diverse analytical tools.
We integrated a multi-detector analytical chromatography with common analytical methods such as ddPCR, infectivity assays, and total DNA and protein analyses. We demonstrate the critical role of process analytical technology to form a comprehensive understanding of the AdV production, and bridge upstream and downstream processes effectively. Our findings reveal that hcDNA significantly affects the capacity of anion exchange columns, underscoring the necessity for a robust pre-chromatographic strategy to mitigate this burden. Despite the differences in clarified lysates from different upstream processes, an anion exchange monolithic column has proven effective, achieving a 2–3 log concentration of adenovirus while eliminating over 99% of protein and DNA impurities.
The findings demonstrate that adopting advanced analytical tools and innovative bioprocessing strategies can significantly improve the quality and scalability of adenovirus production, meeting the demands of both research and commercial applications. This approach not only ensures the production of safer and more effective adenoviral vectors but also offers a cost-effective solution.

Location: Cardiff, Wales