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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
High-level protein expression in scalable CHO transient transfection.
Jianxin Ye1, Vanessa Kober, Melanie Tellers
1Merck & Co., Inc., Bioprocess Research & Development, Rahway, New Jersey 07065, USA. Jianxin_ye@merck.com
Biotechnology and Bioengineering
|February 10, 2009
Summary
We developed a high-yield Chinese hamster ovary (CHO) cell transient transfection platform for rapid therapeutic protein production. This optimized process achieved high monoclonal antibody (Mab) expression levels, suitable for pharmaceutical development.
Area of Science:
- Biotechnology
- Cell Line Development
- Protein Expression
Background:
- Chinese hamster ovary (CHO) cells are industry standard for therapeutic protein production.
- Rapid protein production in early development necessitates efficient transient transfection platforms.
- Current CHO transient transfection methods often lack sufficient protein expression levels.
Purpose of the Study:
- To develop an optimized CHO transient transfection platform for high-level therapeutic protein expression.
- To identify key factors enhancing transfection efficiency and protein yield in CHO cells.
- To demonstrate the scalability and comparability of protein quality from the developed platform.
Main Methods:
- Utilized polyethylenimine (PEI) for transfection in CHO cells.
- Screened various media, identifying UltraCHO as optimal.
- Optimized transfection conditions using DMSO and lithium acetate (LiAc).
- Developed a 14-day fed-batch process for enhanced production yield.
- Scaled the process to a 10 L working volume in a wave bioreactor.
Main Results:
- Achieved an average monoclonal antibody (Mab) expression level of 80 mg/L.
- Demonstrated significant improvements in CHO transient transfection expression using DMSO and LiAc.
- Successfully scaled the transient transfection process to 10 L.
- Mabs produced showed comparable glycosylation patterns to those from stable CHO cell lines.
Conclusions:
- An optimized CHO transient transfection platform enables high-level therapeutic protein production.
- The developed platform offers a rapid and efficient alternative for early-stage biopharmaceutical development.
- Transiently expressed Mabs in CHO cells exhibit similar glycosylation profiles to stably expressed Mabs, unlike those from HEK293EBNA cells.
