Comparative Study of Two Tenecteplase Therapeutic Protein Purification Methods
Therapeutic Tenecteplase (TNKase) is a recombinant and site directed mutant version of human tissue plasminogen activator (TPA) with clinical advantages over TPA. Due to specific glycosylation, TNKase is preferred to over-express in mammalian cell lines such as Chinese Hamster Ovarian (CHO) Cells. T...
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| Format: | Article |
| Language: | English |
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Shiraz University of Medical Sciences
2021-09-01
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| Series: | Trends in Pharmaceutical Sciences |
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| Online Access: | https://tips.sums.ac.ir/article_47722_efe5fa2e3e5a9dd64737d26fb46cd3f7.pdf |
| _version_ | 1818826151006568448 |
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| author | Hossein Javid |
| author_facet | Hossein Javid |
| author_sort | Hossein Javid |
| collection | DOAJ |
| description | Therapeutic Tenecteplase (TNKase) is a recombinant and site directed mutant version of human tissue plasminogen activator (TPA) with clinical advantages over TPA. Due to specific glycosylation, TNKase is preferred to over-express in mammalian cell lines such as Chinese Hamster Ovarian (CHO) Cells. The production and purification of this protein need huge efforts and costs, which directly increase the end product price and limits its medical applications in developing countries despite its benefits. In the current study, we compared two purification methods in order to minimize purification steps as well as purification costs. In the first method, DMEM medium containing CHO-C111 cells expressed recombinant TNKase was purified by a three columns protocol including Sephadex® G-10, HiPrepTM CM FF and L-lysine HyperD®. In the second method, because of its properties, only L-lysine HyperD® column was applied for purification of protein molecules with the lysine binding site, including TNKase. Our results showed that in the second method, higher purification fold and purification yield (1.14 and 1.25 times, respectively) have achieved compared to the first method. This finding in addition to reduction in purification steps, purification cost and time, make it possible to use this method for purification of TNKase. In addition, we suggest overexpressing this protein in serum-free cell lines such as CHO-DG44 in order to minimize impurities and make purification procedure easier. |
| first_indexed | 2024-12-19T00:23:06Z |
| format | Article |
| id | doaj.art-0681fc1f54d44cec897d2abe230e67b4 |
| institution | Directory Open Access Journal |
| issn | 2423-5652 |
| language | English |
| last_indexed | 2024-12-19T00:23:06Z |
| publishDate | 2021-09-01 |
| publisher | Shiraz University of Medical Sciences |
| record_format | Article |
| series | Trends in Pharmaceutical Sciences |
| spelling | doaj.art-0681fc1f54d44cec897d2abe230e67b42022-12-21T20:45:25ZengShiraz University of Medical SciencesTrends in Pharmaceutical Sciences2423-56522021-09-017319120010.30476/tips.2021.91710.110447722Comparative Study of Two Tenecteplase Therapeutic Protein Purification MethodsHossein Javid0Iranian Academic Center for Education, Culture & Research (ACECR)‐Fars Branch Department of Human Genetics Research, Shiraz, IranTherapeutic Tenecteplase (TNKase) is a recombinant and site directed mutant version of human tissue plasminogen activator (TPA) with clinical advantages over TPA. Due to specific glycosylation, TNKase is preferred to over-express in mammalian cell lines such as Chinese Hamster Ovarian (CHO) Cells. The production and purification of this protein need huge efforts and costs, which directly increase the end product price and limits its medical applications in developing countries despite its benefits. In the current study, we compared two purification methods in order to minimize purification steps as well as purification costs. In the first method, DMEM medium containing CHO-C111 cells expressed recombinant TNKase was purified by a three columns protocol including Sephadex® G-10, HiPrepTM CM FF and L-lysine HyperD®. In the second method, because of its properties, only L-lysine HyperD® column was applied for purification of protein molecules with the lysine binding site, including TNKase. Our results showed that in the second method, higher purification fold and purification yield (1.14 and 1.25 times, respectively) have achieved compared to the first method. This finding in addition to reduction in purification steps, purification cost and time, make it possible to use this method for purification of TNKase. In addition, we suggest overexpressing this protein in serum-free cell lines such as CHO-DG44 in order to minimize impurities and make purification procedure easier.https://tips.sums.ac.ir/article_47722_efe5fa2e3e5a9dd64737d26fb46cd3f7.pdftenecteplasepurificationhuman tissue plasminogen activator |
| spellingShingle | Hossein Javid Comparative Study of Two Tenecteplase Therapeutic Protein Purification Methods Trends in Pharmaceutical Sciences tenecteplase purification human tissue plasminogen activator |
| title | Comparative Study of Two Tenecteplase Therapeutic Protein Purification Methods |
| title_full | Comparative Study of Two Tenecteplase Therapeutic Protein Purification Methods |
| title_fullStr | Comparative Study of Two Tenecteplase Therapeutic Protein Purification Methods |
| title_full_unstemmed | Comparative Study of Two Tenecteplase Therapeutic Protein Purification Methods |
| title_short | Comparative Study of Two Tenecteplase Therapeutic Protein Purification Methods |
| title_sort | comparative study of two tenecteplase therapeutic protein purification methods |
| topic | tenecteplase purification human tissue plasminogen activator |
| url | https://tips.sums.ac.ir/article_47722_efe5fa2e3e5a9dd64737d26fb46cd3f7.pdf |
| work_keys_str_mv | AT hosseinjavid comparativestudyoftwotenecteplasetherapeuticproteinpurificationmethods |