| Chapter title |
PEI-Mediated Transient Gene Expression in CHO Cells
|
|---|---|
| Chapter number | 3 |
| Book title |
Recombinant Protein Expression in Mammalian Cells
|
| Published in |
Methods in molecular biology, September 2018
|
| DOI | 10.1007/978-1-4939-8730-6_3 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8729-0, 978-1-4939-8730-6
|
| Authors |
Yashas Rajendra, Rajendra, Yashas |
| Abstract |
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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 17 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 3 | 18% |
| Other | 2 | 12% |
| Researcher | 2 | 12% |
| Student > Master | 2 | 12% |
| Student > Doctoral Student | 1 | 6% |
| Other | 0 | 0% |
| Unknown | 7 | 41% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 7 | 41% |
| Chemical Engineering | 1 | 6% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 6% |
| Neuroscience | 1 | 6% |
| Unknown | 7 | 41% |