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KMID : 0545120130230081116
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Journal of Microbiology and Biotechnology
2013 Volume.23 No. 8 p.1116 ~ p.1122
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Engineering the Cellular Protein Secretory Pathway for Enhancement of Recombinant Tissue Plasminogen Activator Expression in Chinese Hamster Ovary Cells: Effects of CERT and XBP1s Genes
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Azam Rahimpour
Behrouz Vaziri
Reza Moazzami
Leila Nematollahi
Farzaneh Barkhordari
Leila Kokabee
Ahmad Adeli
Fereidoun Mahboudi
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Abstract
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Cell line development is the most critical and also the most time-consuming step in the production of recombinant therapeutic proteins. In this regard, a variety of vector and cell engineering strategies have been developed for generating high-producing mammalian cells; however, the cell line engineering approach seems to show various results on different recombinant protein producer cells. In order to improve the secretory capacity of a recombinant tissue plasminogen activator (t-PA)-producing Chinese hamster ovary (CHO) cell line, we developed cell line engineering approaches based on the ceramide transfer protein (CERT) and X-box binding protein 1 (XBP1) genes. For this purpose, CERT S132A, a mutant form of CERT that is resistant to phosphorylation, and XBP1s were overexpressed in a recombinant t-PA-producing CHO cell line. Overexpression of CERT S132A increased the specific productivity of t-PA-producing CHO cells up to 35%. In contrast, the heterologous expression of XBP1s did not affect the t-PA expression rate. Our results suggest that CERTS132A- based secretion engineering could be an effective strategy for enhancing recombinant t- PA production in CHO cells.
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KEYWORD
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Chinese hamster ovary cells, cell line engineering, X-box binding protein 1, ceramide transfer protein, tissue plasminogen activator
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