Applied Microbiology and Biotechnology

, Volume 102, Issue 15, pp 6469–6477 | Cite as

Process optimization for the rapid production of adenoviral vectors for clinical trials in a disposable bioreactor system

  • Ke-Da Chen
  • Xiao-Xin Wu
  • Dong-Shan Yu
  • Hui-Lin Ou
  • Yan-Hua Li
  • Yu-Qing Zhou
  • Lan-Juan LiEmail author
Biotechnological products and process engineering


Recombinant adenoviral (Ad) vectors are highly efficient gene transfer vectors widely used in vaccine development and immunotherapy. To promote the industrial application of Ad vectors, studies focusing on reducing the cost of manufacturing, shortening the preclinical research period, and improving the quality of products are needed. Here, we describe a highly efficient and economical process for producing Ad vector in a novel, single-use bioreactor system suitable for clinical trials. A mini-bioreactor was used for parameter optimization and development of medium replacement protocols for Ad5-GFP production before scale-up. HEK293 cell culture and virus infection were monitored in a disposable AmProtein Current Perfusion Bioreactor and Bioflo310 bioreactor using optimized parameters and medium replacement protocols. The total cell number increased from 2.0 × 109 to 3.2 × 1010 after 6 days of culture. The total number of viral particles obtained in a single batch was 1.2 × 1015. These results demonstrate the efficiency and suitability of this system for Ad vector production for research and GMP applications.


Adenoviral vector Disposable bioreactor HEK293 cells Process optimization 



We would like to thank AmProtein (Hangzhou, China) for their assistance with ACPB photos. We also thank the native English speaking scientists of Elixigen Company (Huntington Beach, California) for editing our manuscript.


This work was supported by the State Project of Essential Drug Research and Development (grant number 2015ZX09101044) and the Science & Technology Key Program of Zhejiang, China (grant number 2014C03039).

Compliance with ethical standards

This article does not contain any studies with human participants or animals performed by any of the authors.

Conflict of interest

The authors declare that they have no conflicts of interest.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory for Diagnosis and Treatment of Infectious Disease, Collaborative Innovative Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated HospitalZhejiang University School of MedicineHangzhouChina

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