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A novel autoregulated proliferation-controlled production process using recombinant CHO cells
X Mazur1, H M Eppenberger, J E Bailey
1Institute of Biotechnology, ETH Zurich, CH-8093 Zurich, Switzerland.
Biotechnology and Bioengineering
|August 24, 1999
Summary
This study introduces a novel Tet(SWITCH) bioprocess for enhanced heterologous protein production. It uses a self-regulated system to control cell proliferation and protein synthesis, improving yields for pharmaceuticals.
Area of Science:
- Biotechnology
- Molecular Biology
- Bioprocess Engineering
Background:
- Controlled cell proliferation enhances heterologous protein production.
- Existing methods require complex medium changes for phase transitions.
Purpose of the Study:
- To develop a self-regulated, two-phase bioprocess for enhanced protein production.
- To enable a seamless transition from cell growth to production phases without medium exchange.
Main Methods:
- Utilized a multicistronic expression unit with a product gene and a cytostatic gene (p27) under a tetracycline-repressible promoter.
- Implemented a Tet(SWITCH) system leveraging the degradation of tetracycline (tet) and doxycycline (dox).
- Controlled phase transition based on initial cell density and tetracycline concentration.
Main Results:
- Achieved a reliable, self-regulated transition to a proliferation-inhibited production phase.
- Enabled a growth-arrested production phase for at least 7 days with high cell viability.
- Demonstrated enhanced heterologous protein production in a well-defined physiological state.
Conclusions:
- The Tet(SWITCH) process offers a reliable and efficient method for enhanced protein production.
- This adaptable technology is suitable for industrial production of challenging protein pharmaceuticals.
- Eliminates the need for medium exchange, simplifying bioprocess operations.