Journal article
PEI-Mediated Transient Gene Expression in CHO Cells.
- Rajendra Y Laboratory of Cellular Biotechnology (LBTC), École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland. rajendra_yashas@lilly.com.
- 2018-09-23
Published in:
- Methods in molecular biology (Clifton, N.J.). - 2018
CHO cells
Orbital shaking
Polyethyleneimine
Recombinant protein
Transfection
Animals
CHO Cells
Cricetulus
Polyethyleneimine
Recombinant Proteins
Transfection
English
We describe a method for polyethyleneimine (PEI) mediated transient transfection of suspension-adapted Chinese hamster ovary (CHO-DG44) cells for protein expression applicable at scales from 2 mL to 2 L. The method involves transfection at a high cell density (5 × 106 cells/mL) by direct addition of plasmid DNA (pDNA) and PEI to the culture and subsequent incubation at 31 °C with agitation by orbital shaking. This method requires 0.3 mg/L of coding pDNA, 2.7 mg/L of nonspecific (filler) DNA and 15 mg/L of PEI. The production phase is performed at 31 °C in the presence of 0.25% N,N-dimethylacetamide (DMA). We also provide information on culture vessel options, recommended working volumes, and recommended shaking speeds for transfections at scales from 2 mL to 2 L.
- Language
-
- English
- Open access status
- closed
- Identifiers
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- DOI 10.1007/978-1-4939-8730-6_3
- PMID 30242678
- Persistent URL
- https://sonar.ch/global/documents/16455
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