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High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
Published on: December 28, 2015
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Development of stable HEK293T cell pools expressing CSFV E2 protein: A potential antigen expression platform
Yanmin Zhang1, Daoyuan Na1, Weijian Zhang1
1The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Vaccine
|February 1, 2023
Summary
Stable HEK293T cell pools efficiently produce large quantities of classical swine fever virus (CSFV) E2 protein. This robust platform achieves high titers (2 g/L) and maintains stability for rapid antigen production.
Area of Science:
- Biotechnology
- Virology
- Protein Expression
Background:
- Protective antigens like classical swine fever virus (CSFV) E2 protein are crucial for diagnostics and vaccines.
- Large-scale antigen production is essential but challenging with traditional methods.
Purpose of the Study:
- To develop a stable HEK293T cell pool for efficient large-scale production of CSFV E2 protein.
- To optimize E2 protein yield and assess the stability and performance of the cell pools.
Main Methods:
- Lentiviral vector transduction to create stable HEK293T cell pools expressing E2 protein.
- Optimization of signal peptide (SP7) and multiplicity of infection (MOI).
- Evaluation of cell pool stability, and E2 protein production in batch, fed-batch, and semi-perfusion cultures.
Main Results:
- Selected SP7 signal peptide significantly increased E2 protein production.
- High MOI enhanced E2 expression by increasing gene copy numbers.
- Stable cell pools maintained integrity for over 90 days, achieving a maximum E2 titer of 2 g/L in semi-perfusion.
Conclusions:
- Stable HEK293T cell pools offer a robust and efficient platform for rapid recombinant protein production.
- The developed system demonstrates high yield and stability, suitable for diagnostic reagents and subunit vaccines.
- No significant differences in E2 protein aggregation or immunogenicity were observed across different culture methods.
