期刊
PROCESSES
卷 10, 期 6, 页码 -出版社
MDPI
DOI: 10.3390/pr10061064
关键词
BMP2; non-viral gene delivery; recombinant protein production; transient gene expression; process development; EGFP; HEK; CHO
资金
- DFG (Deutsche Forschungsgemeinschaft) [411774929 (FR 830/21-1)]
- DFG
This study demonstrates that transient gene expression (TGE)-based production of BMP2 is more efficient in HEKsus cells than in CHOsus cells. The use of a repeated transient transfection (RTT) process in HEKsus cells significantly improves BMP2 production and extends the production phase.
Human bone morphogenetic protein 2 (hBMP2) is routinely used in medical applications as an inducer of osteoformation. The recombinant production of BMP2 is typically performed using stable Chinese hamster ovary (CHO) cell lines. However, this process is inefficient, resulting in low product titers. In contrast, transient gene expression (TGE), which also enables the production of recombinant proteins, suffers from short production times and hence limited total product amounts. Here, we show that TGE-based BMP2 production is more efficient in HEKsus than in CHOsus cells. Independently of the cell lines, a bicistronic plasmid co-expressing EGFP and BMP2 facilitated the determination of the transfection efficiency but led to inferior BMP2 titers. Finally, we used a high cell density transient transfection (HCD-TGE) protocol to improve and extend the BMP2 expression by performing four rounds of serial transfections on one pool of HEKsus cells. This repeated transient transfection (RTT) process in HEKsus cells was implemented using EGFP as a reporter gene and further adapted for BMP2 production. The proposed method significantly improves BMP2 production (up to 509 ng/10(6) cells) by extending the production phase (96-360 h). RTT can be integrated into the seed train and is shown to be compatible with scale-up to the liter range.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据