title: Transient expression of recombinant sPDGFR alpha-Fc in CHO DG44 cells using 50-mL orbitally shaking disposable bioreactors.
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 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.
authors.affiliation: Biomedical R&D Center, Guangdong Provincial Key Laboratory of Bioengineering Medicine, National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou 510632, Guangdong, PR China.
authors.forename: Yun-Xia
authors.initials: YX
authors.lastname: Sang
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Zhang
authors.forename: Xiao-Jia
authors.initials: XJ
authors.lastname: Chen
authors.forename: Kui
authors.initials: K
authors.lastname: Xie
authors.forename: Chui-Wen
authors.initials: CW
authors.lastname: Qian
authors.forename: An
authors.initials: A
authors.lastname: Hong
authors.forename: Qiu-Ling
authors.initials: QL
authors.lastname: Xie
authors.forename: Sheng
authors.initials: S
authors.lastname: Xiong
mesh_terms: D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002470:Cell Survival; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D015815:Cell Adhesion Molecules; C106607:FCGBP protein, human; D011993:Recombinant Fusion Proteins; D020796:Receptor, Platelet-Derived Growth Factor alpha