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
Protein & Peptide Letters
Title: Transient Expression of Recombinant sPDGFRα-Fc in CHO DG44 Cells using 50-mL Orbitally Shaking Disposable Bioreactors
Volume: 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
Affiliation:
Keywords: PDGF, soluble PDGFRα-Fc, transient gene expression, CHO DG44 cells, Orbitally shaking bioreactor, disposable bioreactor
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.
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Sang Yun-Xia, Zhang Xiao-Wei, Chen Xiao-Jia, Xie Kui, Qian Chui-Wen, Hong An, Xie Qiu-Ling and Xiong Sheng, Transient Expression of Recombinant sPDGFRα-Fc in CHO DG44 Cells using 50-mL Orbitally Shaking Disposable Bioreactors, Protein & Peptide Letters 2010; 17 (7) . https://dx.doi.org/10.2174/092986610791306733
DOI https://dx.doi.org/10.2174/092986610791306733 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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