title: 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.
abstract: 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.
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Tehran 1316943551, Iran.
authors.forename: Azam
authors.initials: A
authors.lastname: Rahimpour
authors.forename: Behrouz
authors.initials: B
authors.lastname: Vaziri
authors.forename: Reza
authors.initials: R
authors.lastname: Moazzami
authors.forename: Leila
authors.initials: L
authors.lastname: Nematollahi
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Barkhordari
authors.forename: Leila
authors.initials: L
authors.lastname: Kokabee
authors.forename: Ahmad
authors.initials: A
authors.lastname: Adeli
authors.forename: Fereidoun
authors.initials: F
authors.lastname: Mahboudi
mesh_terms: D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D004268:DNA-Binding Proteins / Q000235:genetics; D005260:Female; D015870:Gene Expression; D060847:Metabolic Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D055571:Secretory Pathway / Q000235:genetics*; D010959:Tissue Plasminogen Activator / Q000235:genetics / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator