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Updated: Jan 19, 2026
EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR
Published on: January 16, 2016
Process optimization for the rapid production of Enterovirus 71
Xiao-Xin Wu1, Ke-Da Chen1, Da-Zhi Chen1
1State Key Laboratory for Diagnosis and Treatment of Infectious Disease, Collaborative Innovative Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, 310003, China.
This study optimized Enterovirus 71 (EV71) production using Vero cells and bioreactors. The efficient process achieved high viral yields, crucial for developing EV71 vaccines against hand-foot-and-mouth disease (HFMD).
Area of Science:
- Virology
- Biotechnology
- Vaccine Development
Background:
- Enterovirus 71 (EV71) causes hand-foot-and-mouth disease (HFMD).
- Effective EV71 vaccines are essential for public health.
- Current EV71 production methods require optimization for efficiency and cost-effectiveness.
Purpose of the Study:
- To develop a highly efficient and economical large-scale production process for Enterovirus 71 (EV71).
- To optimize cell culture conditions for maximizing EV71 yield.
- To validate the optimized process in a large-scale bioreactor system.
Main Methods:
- EV71 epidemic strain (EV71-2013ZJHFMD) was purified.
- Vero cells were cultured in a mini-bioreactor with polymer fiber carriers.
- Optimization of seeding density, multiplicity of infection (MOI), and temperature was performed.
- The optimized protocol was scaled up in a 10 L perfusion bioreactor (ACPB).
Main Results:
- Optimal conditions identified: seeding density 1.5 × 107 cells/0.6 g carrier, MOI 0.1, temperature 32°C.
- Cell numbers increased from 1.5 × 109 to 3.0 × 1010.
- Maximum viral titer reached 1.0 × 108/mL with a total yield of 1.93 × 1012 virus particles in 25 L supernatant.
Conclusions:
- A highly efficient large-scale EV71 production process using Vero cells on polymer fiber carriers was established.
- The optimized perfusion bioreactor system significantly enhanced EV71 yield.
- This scalable method provides a foundation for cost-effective EV71 vaccine production.
