loading page

A high cell density perfusion process for MVA virus production: process integration with inline DNA digestion and cost analysis
  • +4
  • Gwendal Gränicher,
  • Masoud Babakhani,
  • Sven Göbel,
  • Ingo Jordan,
  • Pavel Marichal-Gallardo,
  • Y Genzel,
  • Udo Reichl
Gwendal Gränicher
Max Planck Institute for Dynamics of Complex Technical Systems
Author Profile
Masoud Babakhani
Max Planck Institute for Dynamics of Complex Technical Systems
Author Profile
Sven Göbel
Max Planck Institute for Dynamics of Complex Technical Systems
Author Profile
Ingo Jordan
ProBioGen AG
Author Profile
Pavel Marichal-Gallardo
Max Planck Institute for Dynamics of Complex Technical Systems
Author Profile
Y Genzel
Max Planck Institute for Dynamics of Complex Technical Systems
Author Profile
Udo Reichl
Max Planck Institute for Dynamics of Complex Technical Systems
Author Profile

Abstract

By integrating continuous cell cultures with continuous purification methods, process yields and product quality attributes were improved over the last 10 years for recombinant protein production. However, for the production of viral vectors such as Modified Vaccinia virus Ankara (MVA), no such studies have been reported although there is an increasing need to meet the requirements for a rising number of clinical trials against infectious or neoplastic diseases. Here, we present for the first time a scalable suspension cell (AGE1.CR.pIX cells) culture-based perfusion process in bioreactors integrating continuous virus harvesting through an acoustic settler with semi-continuous chromatographic purification. This allowed to obtain purified MVA particles with a space-time yield >600% higher for the integrated perfusion process (1.05 x 1011 TCID50/Lbioreactor/day) compared to the integrated batch process. Without further optimization, purification by membrane-based steric exclusion chromatography resulted in an overall product recovery of 50.5%. To decrease the level of host cell DNA prior to chromatography, a novel inline continuous DNA digestion step was integrated into the process train. A detailed cost analysis comparing integrated production in batch versus production in perfusion mode showed that the cost per dose for MVA was reduced by nearly one third using this intensified small-scale process.

Peer review status:UNDER REVIEW

23 Mar 2021Submitted to Biotechnology and Bioengineering
23 Mar 2021Assigned to Editor
23 Mar 2021Submission Checks Completed
10 Apr 2021Reviewer(s) Assigned
30 May 2021Editorial Decision: Revise Major
30 May 2021Review(s) Completed, Editorial Evaluation Pending