Expression of a novel chimeric truncated t-PA in CHO cells based on in silico experiments.

Fatemeh Davami 1 Soroush Sardari 2 Keivan Majidzadeh-A 1 Mehdi Hemayatkar 1 Farzaneh Barkhrdari 1 Maryam Omidi 1 Mehrnaz Azami 1 Ahmad Adeli 1 Noushin Davoudi 1 Fereidoun Mahboudi 1, *
* Corresponding author
1 Biotechnology Research Center
2 Drug Design and Bioinformatics Unit
Abstract : Tissue plasminogen activator (t-PA) is one of the fibrin-specific serine proteases that play a crucial role in the fibrinolytic system. The rapid clearance of the drug from the circulation, caused by its active uptake in the liver, has lead to complicated clinical applications. Different forms of plasminogen activators have been developed to treat thrombotic disease. Deletion of the first three domains of t-PA by gene manipulation techniques has shown a significant increase in its plasma half life. In order to compensate the disadvantage of higher bleeding risk, a novel chimeric truncated form of t-PA with 394 amino acids and more fibrin affinity compared to the truncated form was designed to be expressed in Chinese Hamster Ovarian (CHO) cells. The recombinant chimeric plasminogen activator consists of kringle 2 and serine protease (K2S) domains of t-PA, namely GHRP-SYQ-K2S. The level of expression was found to be 752 IU/ml with 566,917 IU/mg specific activity, based on amidolytic activity. The fibrin binding of this novel chimeric truncated t-PA was 86% of the full length t-PA at a fibrinogen concentration of 0.2 mg/ml. This could be a promising approach with more desirable pharmacodynamic properties compared to existing commercial forms.
Keywords : Truncated t-PA CHO Cells in Silico Experiments
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Fatemeh Davami, Soroush Sardari, Keivan Majidzadeh-A, Mehdi Hemayatkar, Farzaneh Barkhrdari, et al.. Expression of a novel chimeric truncated t-PA in CHO cells based on in silico experiments.. Journal of Biomedicine and Biotechnology, Hindawi Publishing Corporation, 2010, 2010, pp.108159. ⟨10.1155/2010/108159⟩. ⟨pasteur-00821614⟩

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