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Transient Expression of Recombinant sPDGFRα-Fc in CHO DG44 Cells using 50-mL Orbitally Shaking Disposable Bioreactors

[ Vol. 17 , Issue. 7 ]

Author(s):

Yun-Xia Sang, Xiao-Wei Zhang, Xiao-Jia Chen, Kui Xie, Chui-Wen Qian, An Hong, Qiu-Ling Xie and Sheng Xiong   Pages 919 - 924 ( 6 )

Abstract:


Overactivity of platelet-derived growth factor (PDGF) has been linked to malignant cancers. High levels of PDGF result in the activation of its receptors (PDGFRs) and the over-proliferation of cells. Therefore, interfering with this signaling pathway in cancer cells could be significant for anti-cancer drug development. In a previous study, the sPDGFRα-Fc fusion protein expressed in static CHO-k1 cells showed an anti-proliferative effect on vascular endothelial cells. However, it was difficult to obtain a large quantity of this fusion protein for further functional studies. In the present study, the sPDGFRα-Fc fusion protein was transiently expressed in Chinese Hamster Ovary (CHO) DG44 cells in 50-mL orbital shaking bioreactors. sPDGFRα-Fc was expressed as a 250-kDa dimeric protein with potential glycosylation. The final yield of sPDGFRα-Fc in the culture supernatant was as high as 16.68 mg/L. Our results suggest that transient expression in orbital shaking bioreactors may be feasible for preparation of recombinant proteins used for preclinical studies.

Keywords:

PDGF, soluble PDGFRα-Fc, transient gene expression, CHO DG44 cells, Orbitally shaking bioreactor, disposable bioreactor

Affiliation:

Biomedical R Center, 5/F Building of Biology, Jinan University, Guangzhou 510632, Guangdong, P.R. China.



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