Efficient mAb production in CHO cells incorporating PEI-mediated transfection, mild hypothermia and the co-expression of XBP-1
✍ Scribed by Joe Codamo; Jeff Jia Cheng Hou; Benjamin S. Hughes; Peter P. Gray; Trent P. Munro
- Book ID
- 102882488
- Publisher
- Wiley (John Wiley & Sons)
- Year
- 2011
- Tongue
- English
- Weight
- 622 KB
- Volume
- 86
- Category
- Article
- ISSN
- 0268-2575
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
BACKGROUND: Transient gene expression (TGE) provides a rapid way to generate recombinant protein biologics for pre‐clinical assessment. Human embryonic kidney (HEK293) cells have traditionally been used for TGE; however, there is demand from industry for efficient, high‐producing TGE systems that utilize Chinese hamster ovary (CHO) cells. A polyethyleneimine (PEI) ‐based TGE process has been developed for CHO cells using an episomal expression system to generate enhanced recombinant protein titers.
RESULTS: A five‐fold improvement in monoclonal antibody (mAb) volumetric productivity was achieved by examining key parameters including transfection medium, cell density, transfection reagent, DNA:reagent ratio, the time of transfer to mild hypothermia and feeding strategy post‐transfection. The Epi‐CHO system allowed for a six‐fold expansion in culture volume post‐transfection without significantly affecting specific productivity. This system generates 400% more mAb per µg of plasmid DNA when compared with a non‐episomal system. In addition, the use of X‐box binding protein 1 to enhance secretion capacity and provide further improvements in mAb production with TGE was investigated.
CONCLUSION: Through optimization of key parameters, our results demonstrate the development of a low‐cost, high‐yielding, episomal TGE system that may be adopted during pre‐clinical biologic drug development. Copyright © 2011 Society of Chemical Industry