Pseudorabies virus production using a serum-free medium in fixed-bed bioreactors with low cell inoculum density
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ORIGINAL RESEARCH PAPER
Pseudorabies virus production using a serum-free medium in fixed-bed bioreactors with low cell inoculum density Jianqi Nie . Yang Sun . Feng Peng . Fei Han . Yankun Yang . Xiuxia Liu . Chunli Liu . Ye Li . Zhonghu Bai
Received: 23 April 2020 / Accepted: 13 August 2020 Ó Springer Nature B.V. 2020
Abstract Fixed-bed bioreactors packed with macrocarriers show great potential to be used for vaccine process development and large-scale production due to distinguishing features of low shear force, high cell adhering surface area, and easy replacement of culture media in situ. As an initial step of utilizing this type of bioreactors for Pseudorabies virus production (PRV) by African green monkey kidney (Vero) cells, we developed a tube-fixed-bed bioreactor in the previous study, which represents a scale-down model for Jianqi Nie and Yang Sun contributed equally to this work. J. Nie F. Peng F. Han Y. Yang The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China
further process optimization. By using this scaledown model, here we evaluated impacts of two strategies (use of serum-free medium and low cell inoculum density) on PRV production, which have benefits of simplifying downstream process and reducing risk of contamination. We first compared Vero cell cultures with different media, bioreactors and inoculum densities, and conclude that cell growth with serum-free medium is comparable to that with serum-containing medium in tube-fixed-bed bioreactor, and low inoculum density supports cell growth only in this bioreactor. Next, we applied serum-free X. Liu C. Liu Z. Bai Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, Wuxi 214122, China
J. Nie F. Peng F. Han Y. Yang X. Liu C. Liu Y. Li (&) Z. Bai (&) National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China e-mail: [email protected] Z. Bai e-mail: [email protected] Y. Sun Institute of Bioengineering, School of Life Sciences, Henan University, Kaifeng 475004, China Y. Yang Z. Bai The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China
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Biotechnol Lett
medium and low inoculum cell density for PRV production. By optimization of time of infection (TOI), multiplicity of infection (MOI) and the harvesting strategy, we obtained total amount of virus particles * 9 log10 TCID50 at 5 days post-infection (dpi) in the tube-fixed-bed bioreactor. This process was then scaled up by 25-fold to a Xcell 1-L fixed-bed bioreactor, which yields totally virus particles of 10.5 log10 TCID50, corresponding to * 3 9 105 doses of vaccine. The process studied in this work holds promise to be developed as a generic platform for the production of vaccines for animal and human health. Keywords Fixed-bed biorea
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