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Iranian Biomedical Journal، جلد ۲۳، شماره ۴، صفحات ۲۷۲-۲۷۹
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عنوان انگلیسی Enhancement of Expression Level of Modified t-PA (TNKase) in Leishmania tarentolae by Induction System
چکیده انگلیسی مقاله Background: The expression of bio-therapeutic proteins in mammalian cells, such as CHO, attains high homogeneity related to post-translational modifications. Although CHO remains the most popular cell line for bestselling biotherapeutic proteins on the market, there are still drawbacks such as expensive culture media, long time line, and high drug cost. Recently, researches on a novel Leishmania protozoan system have confirmed that this low level eukaryote could represent a competitive alternative to the mammalian cell lines. Methods: The full length of coding sequence of modified tPA TNKase tenecteplase) was synthesized and cloned into an inducible expression vector of L. tarentolae T7-TR cells. Results: The expression of the construct was driven by a Tet-inducible promoter. A Leishmania secretory signal sequence was also added to the expression cassette to facilitate the release of the recombinant protein into the medium. The secretory recombinant protein was analyzed and confirmed by SDS-PAGE and Western blot analyses. The expression level of TNKase in this novel system of L. tarentolae was 810 IU/mL after induction, which means that the percentage of expression increases two times compared to previous models in L. tarentolae. The TNKase activity was comparable with Activase. Conclusion: Our results suggested that expressed TNK (modified tPA) is functionally compatible with Activase regarding their effect on fibrinolysis. Given the post-translational modification similarities between mammalian and L. tarentolae, it is speculated that this system is capable of producing complex proteins such as tPA similar to mammalian system, with easier manipulation and non-expensive method.
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نویسندگان مقاله | Mohammad Mehdi Attarpour Yazdi
Department of Microbiology, Faculty of Medicine, Shahed University, Tehran, Iran


| Nikky Tofighi
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran


| Taraneh Rajaee
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran


| Marzieh Ghahremanlou
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran


| Ahmad Adeli
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran


| Azam Bolhassani
Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran


| Mohammad Azizi
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran


| Noushin Davoudi
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
نشانی اینترنتی http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-751&slc_lang=en&sid=1
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/125/article-125-1498983.pdf
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زبان مقاله منتشر شده en
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