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Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments
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 pl...
Autores principales: | , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Hindawi Publishing Corporation
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2946600/ https://www.ncbi.nlm.nih.gov/pubmed/20885932 http://dx.doi.org/10.1155/2010/108159 |
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author | Davami, Fatemeh Sardari, Soroush Majidzadeh-A, Keivan Hemayatkar, Mehdi Barkhrdari, Farzaneh Omidi, Maryam Azami, Mehrnaz Adeli, Ahmad Davoudi, Noushin Mahboudi, Fereidoun |
author_facet | Davami, Fatemeh Sardari, Soroush Majidzadeh-A, Keivan Hemayatkar, Mehdi Barkhrdari, Farzaneh Omidi, Maryam Azami, Mehrnaz Adeli, Ahmad Davoudi, Noushin Mahboudi, Fereidoun |
author_sort | Davami, Fatemeh |
collection | PubMed |
description | 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. |
format | Text |
id | pubmed-2946600 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-29466002010-09-30 Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments Davami, Fatemeh Sardari, Soroush Majidzadeh-A, Keivan Hemayatkar, Mehdi Barkhrdari, Farzaneh Omidi, Maryam Azami, Mehrnaz Adeli, Ahmad Davoudi, Noushin Mahboudi, Fereidoun J Biomed Biotechnol Research Article 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. Hindawi Publishing Corporation 2010 2010-09-22 /pmc/articles/PMC2946600/ /pubmed/20885932 http://dx.doi.org/10.1155/2010/108159 Text en Copyright © 2010 Fatemeh Davami et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Davami, Fatemeh Sardari, Soroush Majidzadeh-A, Keivan Hemayatkar, Mehdi Barkhrdari, Farzaneh Omidi, Maryam Azami, Mehrnaz Adeli, Ahmad Davoudi, Noushin Mahboudi, Fereidoun Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments |
title | Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments |
title_full | Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments |
title_fullStr | Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments |
title_full_unstemmed | Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments |
title_short | Expression of a Novel Chimeric Truncated t-PA in CHO Cells Based on in Silico Experiments |
title_sort | expression of a novel chimeric truncated t-pa in cho cells based on in silico experiments |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2946600/ https://www.ncbi.nlm.nih.gov/pubmed/20885932 http://dx.doi.org/10.1155/2010/108159 |
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