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Optimization of an adenovirus-vectored zoster vaccine production process with chemically defined medium and a perfusion system

Biotechnology Letters, ISSN: 1573-6776, Vol: 44, Issue: 11, Page: 1347-1358
2022
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Article Description

Objectives: Cells grown in chemically defined medium are sensitive to shear force, potentially resulting in decreased cell growth. We optimized the perfusion process for HEK293 cell-based recombinant adenovirus-vectored zoster vaccine (Ad-HER) production with chemically defined medium. Methods: We first studied the pseudo-continuous strategies in shake flasks as a mimic of the bioreactor equipped with perfusion systems. Using design of experiment (DoE) in shake flasks, we obtained the regression models between Ad-HER titer/virus input–output ratio and three production process parameters: time of infection (TOI), multiplicity of infection (MOI), and virus production pH (pH). We then confirmed the effect of Pluronic F68 (PF-68) at 3.0 g/L on HEK293 cell growth and Ad-HER production in shake flasks and a 2 L benchtop bioreactor. Results: The optimized process was scale-up to a 2 L benchtop bioreactor with the PATFP perfusion system, which yielded cell density of 7.4 × 10 cells/mL and Ad-HER titer of 9.8 × 10 IFU/mL at 2 dpi, comparable to the bioreactor with a ATF2 system. Conclusion: This optimization strategy could be used to develop a robust process with stable cell culture performance and adenovirus titer. Increasing PF-68 concentration in chemically defined medium could protect cells from shear stress generated by perfusion system.

Bibliographic Details

Nie, Jianqi; Sun, Yang; Ren, He; Huang, Lingling; Feng, Kai; Li, Ye; Bai, Zhonghu

Springer Science and Business Media LLC

Biochemistry, Genetics and Molecular Biology; Chemical Engineering; Immunology and Microbiology

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