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http://dx.doi.org/10.4014/jmb.1106.05060

Combined TGE-SGE Expression of Novel PAI-1-Resistant t-PA in CHO DG44 Cells Using Orbitally Shaking Disposable Bioreactors  

Davami, Fatemeh (Biotechnology Research Center, Pasteur Institute of Iran)
Barkhordari, Farzaneh (Biotechnology Research Center, Pasteur Institute of Iran)
Alebouyeh, Mahmoud (Biotechnology Research Center, Pasteur Institute of Iran)
Adeli, Ahmad (Biotechnology Research Center, Pasteur Institute of Iran)
Mahboudi, Fereidoun (Biotechnology Research Center, Pasteur Institute of Iran)
Publication Information
Journal of Microbiology and Biotechnology / v.21, no.12, 2011 , pp. 1299-1305 More about this Journal
Abstract
An important modification of thrombolytic agents is resistance to plasminogen activator inhibitor-1 (PAI-1). In previous studies, a new truncated PAI-1-resistant variant was developed based on deletion of the first three domains in t-PA and the substitution of KHRR 128-131 amino acids with AAAA in the truncated t-PA. The novel variant expressed in a static culture system of Chinese Hamster Ovary (CHO) DG44 cells exhibited a higher resistance to PAI-1 when compared with the full-length commercial drug; Actylase. In the present study, the truncated-mutant protein was expressed in CHO DG44 cells in 50 ml orbital shaking bioreactors. The final yield of the truncated-mutant in the culture was 752 IU/ml, representing a 63% increase compared with the static culture system. Therefore, these results suggest that using the combined features of a transient and stable expression system is feasible for the production of novel recombinant proteins in the quantities needed for preclinical studies.
Keywords
Tissue plasminogen activator (t-PA); Chinese Hamster Ovary (CHO); suspension culture; orbital shaker; plasminogen activator inhibitor-1 (PAI-1);
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