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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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Transient Gene Expression in Suspension HEK293-EBNA1 Cells
Denis L'Abbé1, Louis Bisson1, Christian Gervais1
1Human Health Therapeutics Research Center, National Research Council Canada, 6100 Royalmount Avenue, Montreal, H4P 2R2, Quebec, Canada.
Methods in Molecular Biology (Clifton, N.J.)
|September 23, 2018
Summary
Transient gene expression in human embryo kidney 293 (HEK293) cells offers a fast method for producing recombinant proteins. This process yields milligram to gram quantities within 10 days using serum-free media and polyethylenimine.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Transient gene expression in HEK293 cells is a common method for rapid recombinant protein production.
- This technique allows for the generation of milligram to gram quantities of proteins within 10 days post-transfection.
Purpose of the Study:
- To describe a simple and robust transfection protocol for suspension-growing HEK293 cells.
- To demonstrate the production and purification of recombinant proteins, including a his-tagged protein and two monoclonal antibodies.
Main Methods:
- Utilizing suspension-growing human embryo kidney 293 (HEK293) cells.
- Employing two commercially available serum-free media.
- Using polyethylenimine (PEI) as the transfection reagent.
Main Results:
- Successful transient gene expression achieved in HEK293 cells.
- Demonstrated production of milligram to gram quantities of recombinant proteins.
- Purification protocols for a his-tagged protein and two monoclonal antibodies were exemplified.
Conclusions:
- A simple and robust transfection method for HEK293 cells using serum-free media and PEI is presented.
- This approach enables efficient and rapid production of recombinant proteins and antibodies.
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