Abstract
Objective
This study was to evaluate the feasibility of using a rocking type bioreactor system, specifically the WAVE 25, in an intensified perfusion culture (IPC) mode for monoclonal antibody (mAb) production in Chinese hamster ovary (CHO) cell line.
Methods
A disposable perfusion bag with floating membrane was used in the IPC process. An automated filter switching system was employed to continuously clarify the harvested post-membrane culture fluid. The overall cell culture performance, product titer, and quality were compared to those of a typical IPC conducted in a bench-top glass bioreactor.
Results
The results showed that the overall trends of cell culture performance, product titer (accumulated harvest volumetric titer) were similar to those of the typical IPC conducted in the glass bioreactor, while the purity related quality were slightly better than the typical run. Furthermore, with the automated filter switching system, the harvested post-membrane culture fluid could be continuously clarified, making it suitable for downstream continuous chromatography.
Conclusion
The study demonstrated the feasibility of using the WAVE-based rocking type bioreactor in the N stage IPC process, which increases the flexibility in adopting IPC process. The results suggest that the rocking type bioreactor system could be a viable alternative to traditional stirred tank bioreactors for perfusion culture in the biopharmaceutical industry.
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- CHO:
-
Chinese hamster ovary
- ATF:
-
Alternating tangential flow
- TFF:
-
Tangential flow filtration
- CSPR:
-
Cell specific perfusion rate
- VCD:
-
Viable cell density
- LPM:
-
Liter per minute
- DO:
-
Dissolved oxygen
- HPLC:
-
High performance liquid chromatography
- HMW:
-
High molecular weight
- IPC:
-
Intensified perfusion culture
- mAb:
-
Monoclonal antibody
- MFC:
-
Mass flow controller
- NTU:
-
Nephelometric turbidity unit
- PC:
-
Principal component
- PCA:
-
Principal component analysis
- Pv :
-
Volumetric productivity
- RCF:
-
Relative centrifugal force
- RPM:
-
Revolutions per minute
- VVM:
-
Air sparger volume per minute/working volume
- SEC-HPLC:
-
Size-exclusion chromatography-high performance liquid chromatography
- CIEF:
-
Capillary isoelectric focusing
- PCC:
-
Periodic counter-current chromatography
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Acknowledgements
We thank Dr. Yifeng Li for his critical reading of the manuscript. We also thank Dr. Weichang Zhou and Dr. Gang Huang for lending their support to this work. The authors would like to thank Downstream Process Development (DSPD) Department, Analytical Science (AS) Department for their assistance with one-step purification and PQA analysis and the media development team of Cell Culture Process Development (CCPD) Department for the media powder used in this work.
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ZL: conceptualization, data curation, formal analysis, methodology, writing—original draft, writing—review & editing. SY: data curation, writing—review & editing. QX: data curation, writing—review & editing. GC: conceptualization, data curation, formal analysis, investigation, methodology, writing—original draft, writing—review & editing.
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Lang, Z., Yan, S., Xiong, Q. et al. WAVE-based intensified perfusion cell culture for fast process development. Biotechnol Lett 45, 1117–1131 (2023). https://doi.org/10.1007/s10529-023-03405-8
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DOI: https://doi.org/10.1007/s10529-023-03405-8