Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents
Purposes: Solubilization of inclusion bodies expressed in E. coli is a critical step during manufacturing of recombinant proteins expressed as inclusion bodies. So far, various methods have been used for solubilization and purification of inclusion body proteins to obtain active proteins with high p...
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| Format: | Article |
| Language: | English |
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Tabriz University of Medical Sciences
2020-02-01
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| Series: | Advanced Pharmaceutical Bulletin |
| Subjects: | |
| Online Access: | https://apb.tbzmed.ac.ir/PDF/apb-27814 |
| _version_ | 1828555098692780032 |
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| author | Samira Nekoufar Ahmad Fazeli Mohammad Reza Fazeli |
| author_facet | Samira Nekoufar Ahmad Fazeli Mohammad Reza Fazeli |
| author_sort | Samira Nekoufar |
| collection | DOAJ |
| description | Purposes: Solubilization of inclusion bodies expressed in E. coli is a critical step during manufacturing of recombinant proteins expressed as inclusion bodies. So far, various methods have been used for solubilization and purification of inclusion body proteins to obtain active proteins with high purity and yield. The aim of this study was to examine the benefit of organic solvents such as alcohols in solubilization of recombinant interferon β-1b inclusion bodies. Methods: Effect of important parameters inclusion pH, concentration and type of denaturant and concentration of alcoholic solvents were optimized to formulate a suitable solubilization buffer and investigate their effect on solubilization of interferon β-1b inclusion bodies. Results: Our findings showed the acidic pH in the range of 2-3 is more suitable than alkaline pH >12 for solubilization and achieving higher content of interferon β-1beta and pure recombinant protein. We have also demonstrated that 1% SDS acts better than 2M urea to solubilize Inclusion body proteins of interferon β-1b at pH of 2-3. The interferon concentration was 2.35 mg per 100 mg IB when we used 40% (v/v) 1-propanol and 20% (v/v) 2-butanol into the buffer solution as well. Conclusion: The optimized method provides gentile condition for solubilization of inclusion body at high protein concentration and purity with a degree of retention of native secondary structure which makes this method valuable to be used in production and research area. |
| first_indexed | 2024-12-12T05:47:30Z |
| format | Article |
| id | doaj.art-b426425253a349e08cabc99bfd3806ca |
| institution | Directory Open Access Journal |
| issn | 2228-5881 2251-7308 |
| language | English |
| last_indexed | 2024-12-12T05:47:30Z |
| publishDate | 2020-02-01 |
| publisher | Tabriz University of Medical Sciences |
| record_format | Article |
| series | Advanced Pharmaceutical Bulletin |
| spelling | doaj.art-b426425253a349e08cabc99bfd3806ca2022-12-22T00:35:45ZengTabriz University of Medical SciencesAdvanced Pharmaceutical Bulletin2228-58812251-73082020-02-0110223323810.34172/apb.2020.027apb-27814Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic SolventsSamira Nekoufar0Ahmad Fazeli1Mohammad Reza Fazeli2Department of Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Sciences (TUMS), Tehran, Iran.Department of Drug & Food Control, The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Sciences, Tehran, Iran.Purposes: Solubilization of inclusion bodies expressed in E. coli is a critical step during manufacturing of recombinant proteins expressed as inclusion bodies. So far, various methods have been used for solubilization and purification of inclusion body proteins to obtain active proteins with high purity and yield. The aim of this study was to examine the benefit of organic solvents such as alcohols in solubilization of recombinant interferon β-1b inclusion bodies. Methods: Effect of important parameters inclusion pH, concentration and type of denaturant and concentration of alcoholic solvents were optimized to formulate a suitable solubilization buffer and investigate their effect on solubilization of interferon β-1b inclusion bodies. Results: Our findings showed the acidic pH in the range of 2-3 is more suitable than alkaline pH >12 for solubilization and achieving higher content of interferon β-1beta and pure recombinant protein. We have also demonstrated that 1% SDS acts better than 2M urea to solubilize Inclusion body proteins of interferon β-1b at pH of 2-3. The interferon concentration was 2.35 mg per 100 mg IB when we used 40% (v/v) 1-propanol and 20% (v/v) 2-butanol into the buffer solution as well. Conclusion: The optimized method provides gentile condition for solubilization of inclusion body at high protein concentration and purity with a degree of retention of native secondary structure which makes this method valuable to be used in production and research area.https://apb.tbzmed.ac.ir/PDF/apb-27814interferon beta-1binclusion bodysolubilizationorganic solventalcohol |
| spellingShingle | Samira Nekoufar Ahmad Fazeli Mohammad Reza Fazeli Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents Advanced Pharmaceutical Bulletin interferon beta-1b inclusion body solubilization organic solvent alcohol |
| title | Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents |
| title_full | Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents |
| title_fullStr | Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents |
| title_full_unstemmed | Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents |
| title_short | Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents |
| title_sort | solubilization of human interferon β 1b inclusion body proteins by organic solvents |
| topic | interferon beta-1b inclusion body solubilization organic solvent alcohol |
| url | https://apb.tbzmed.ac.ir/PDF/apb-27814 |
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