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، جلد ۶، شماره ۲، صفحات ۲۹-۳۵
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| عنوان فارسی |
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| چکیده فارسی مقاله |
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| کلیدواژه‌های فارسی مقاله |
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| عنوان انگلیسی |
Expression of Recombinant Factor IX Using the Transient Gene Expression Technique |
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| چکیده انگلیسی مقاله |
Background: Pilot and large-scale production of recombinant proteins require the presence of stable clones, but the process of selecting stable clones is time consuming. Moreover, continuous clone culturing in large-scale production may cause loss of incoming plasmid and recombinant genes. Considering the advancements in Transient Gene Expression (TGE) technology, the large-scale expression of factor IX (FIX) was investigated in HEK cells by the TGE technique. Materials and methods: HEK cells were seeded in a cell factory, and then transfected by pcDNA-hFIX plasmid using calcium phosphate co-precipitation method. Stable HEK-hFIX cells were also seeded in a cell factory, separately. After adding vitamin K, recombinant FIX was quantified in conditioned media using an ELISA. Moreover, its functional activity was assayed using an aPTT test. Results: The results showed that the expression and activity of FIX by TGE technology was, respectively, 1.6 and 1.5 times higher than that obtained through stable HEK-FIX cells. Since calculating the specific activity revealed that for all time periods it is 0.2 mU/ng, so the increase in activity is due to the increase in the amount of FIX. Conclusions: HEK cells with higher transfectability seemed to be an appropriate alternative for transient expression for large-scale protein production. Furthermore, if rapid expression of recombinant proteins is intended, TGE can replace costly and low-yield methods. |
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| کلیدواژه‌های انگلیسی مقاله |
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| نویسندگان مقاله |
جعفر وطن دوست | Jafar Vatandoost
Department of Biology, Hakim Sabzevari University, Sabzevar
دانشگاه حکیم سبزواری
محمدامین عظیم فر | Mohammad amin Azimifar
Department of Biotechnology, Sabzevar Branch, Islamic Azad University, Sabzevar, Iran
دانشگاه آزاد سبزوار
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| نشانی اینترنتی |
http://rmm.mazums.ac.ir/browse.php?a_code=A-10-1050-1&slc_lang=en&sid=1 |
| فایل مقاله |
اشکال در دسترسی به فایل - ./files/site1/rds_journals/2735/article-2735-2201160.pdf |
| کد مقاله (doi) |
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| زبان مقاله منتشر شده |
en |
| موضوعات مقاله منتشر شده |
بیوتکنولوژی |
| نوع مقاله منتشر شده |
پژوهشی |
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