Combined TGE-SGE expression of novel PAI-1-resistant t-PA in CHO DG44 cells using orbitally shaking disposable bioreactors.
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 truncatedmutant protein was expressed in CHO DG44 cells in 50 ml orbital shaking bioreactors. The final yield of the truncatedmutant 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.
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Fatemeh
F
Davami
Farzaneh
F
Barkhordari
Mahmoud
M
Alebouyeh
Ahmad
A
Adeli
Fereidoun
F
Mahboudi
D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D006224:Cricetinae; D015870:Gene Expression; D006801:Humans; D017395:Plasminogen Activator Inhibitor 1 / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D017395:Plasminogen Activator Inhibitor 1; D011993:Recombinant Fusion Proteins; D010959:Tissue Plasminogen Activator