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

Sang Yun Xia; Zhang Xiao Wei; Chen Xiao Jia; Xie Kui; Qian Chui Wen; Hong An; Xie Qiu Ling*; Xiong Sheng
SCI
南昌大学; 济南大学; 暨南大学

摘要

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 sPDGFR alpha-Fc fusion protein expressed in static CHO-k(1) 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 sPDGFR alpha-Fc fusion protein was transiently expressed in Chinese Hamster Ovary (CHO) DG44 cells in 50-mL orbital shaking bioreactors. sPDGFR alpha-Fc was expressed as a 250-kDa dimeric protein with potential glycosylation. The final yield of sPDGFR alpha-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.

关键词

PDGF soluble PDGFR alpha-Fc transient gene expression CHO DG44 cells Orbitally shaking bioreactor disposable bioreactor