Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion Protein
Background: The N-terminal domain of the myelin oligodendrocyte glycoprotein (MOG) has been shown to generate experimental autoimmune encephalomyelitis (EAE), an animal model of MS. A considerable amount of MOG must be accessible for EAE induction. Here, for the first time, Response Surface Methodol...
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| Format: | Article |
| Language: | English |
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Tabriz University of Medical Sciences
2024-04-01
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| Series: | Pharmaceutical Sciences |
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| Online Access: | https://ps.tbzmed.ac.ir/PDF/ps-30-252.pdf |
| _version_ | 1797209886535188480 |
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| author | Maryam Radmard Atieh Hashemi |
| author_facet | Maryam Radmard Atieh Hashemi |
| author_sort | Maryam Radmard |
| collection | DOAJ |
| description | Background: The N-terminal domain of the myelin oligodendrocyte glycoprotein (MOG) has been shown to generate experimental autoimmune encephalomyelitis (EAE), an animal model of MS. A considerable amount of MOG must be accessible for EAE induction. Here, for the first time, Response Surface Methodology-Box-Behnken (RSM-BBD) was employed to identify the ideal culture conditions for causing Escherichia coli (E. coli) BL21 to overproduce the Thioredoxin-MOG (Trx-MOG) fusion protein. The RSM method is a powerful, efficient, and reliable alternative to the One-Factor-At-A-Time (OFAT) method in optimizing process variables, allowing for a smaller number of experimental runs, investigating variable interaction, and being cheaper and less time-consuming. Methods: Here, using the 29 experimental assays, the direct and indirect effects of factors including post-induction time, IPTGinducer concentration, pre-induction optical density, and post-induction temperature on the protein expression level content were evaluated. Results: The proposed quadratic model demonstrated a significant effect of the two variables A (time) and C (temperature) on protein synthesis. An inducer concentration of 0.491 mM, the pre-induction optical density (OD600) of 0.8, and a temperature of 23 °C for 23.878 hours were found to be the best growth conditions for high yield Trx-MOG synthesis. The optimum protein concentration was attained (163.96 µg/mL) and was within the range of (200.04 µg/mL), which was the value predicted. Conclusion: The study concluded that RSM optimization effectively increased the production of Trx-MOG in E. coli, which could have the potential for large-scale fermentation. |
| first_indexed | 2024-04-24T10:01:49Z |
| format | Article |
| id | doaj.art-3f0821bbf3eb498f8e2010efc2d04d9c |
| institution | Directory Open Access Journal |
| issn | 2383-2886 |
| language | English |
| last_indexed | 2024-04-24T10:01:49Z |
| publishDate | 2024-04-01 |
| publisher | Tabriz University of Medical Sciences |
| record_format | Article |
| series | Pharmaceutical Sciences |
| spelling | doaj.art-3f0821bbf3eb498f8e2010efc2d04d9c2024-04-13T10:40:37ZengTabriz University of Medical SciencesPharmaceutical Sciences2383-28862024-04-0130225226110.34172/PS.2024.1ps-40000Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion ProteinMaryam Radmard0Atieh Hashemi1Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.Background: The N-terminal domain of the myelin oligodendrocyte glycoprotein (MOG) has been shown to generate experimental autoimmune encephalomyelitis (EAE), an animal model of MS. A considerable amount of MOG must be accessible for EAE induction. Here, for the first time, Response Surface Methodology-Box-Behnken (RSM-BBD) was employed to identify the ideal culture conditions for causing Escherichia coli (E. coli) BL21 to overproduce the Thioredoxin-MOG (Trx-MOG) fusion protein. The RSM method is a powerful, efficient, and reliable alternative to the One-Factor-At-A-Time (OFAT) method in optimizing process variables, allowing for a smaller number of experimental runs, investigating variable interaction, and being cheaper and less time-consuming. Methods: Here, using the 29 experimental assays, the direct and indirect effects of factors including post-induction time, IPTGinducer concentration, pre-induction optical density, and post-induction temperature on the protein expression level content were evaluated. Results: The proposed quadratic model demonstrated a significant effect of the two variables A (time) and C (temperature) on protein synthesis. An inducer concentration of 0.491 mM, the pre-induction optical density (OD600) of 0.8, and a temperature of 23 °C for 23.878 hours were found to be the best growth conditions for high yield Trx-MOG synthesis. The optimum protein concentration was attained (163.96 µg/mL) and was within the range of (200.04 µg/mL), which was the value predicted. Conclusion: The study concluded that RSM optimization effectively increased the production of Trx-MOG in E. coli, which could have the potential for large-scale fermentation.https://ps.tbzmed.ac.ir/PDF/ps-30-252.pdfexperimental autoimmune encephalomyelitismultiple sclerosismyelin oligodendrocyte glycoproteinresponse surface methodology |
| spellingShingle | Maryam Radmard Atieh Hashemi Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion Protein Pharmaceutical Sciences experimental autoimmune encephalomyelitis multiple sclerosis myelin oligodendrocyte glycoprotein response surface methodology |
| title | Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion Protein |
| title_full | Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion Protein |
| title_fullStr | Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion Protein |
| title_full_unstemmed | Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion Protein |
| title_short | Response Surface Methodology Approach to Optimize the Expression of Thioredoxin-MOG Fusion Protein |
| title_sort | response surface methodology approach to optimize the expression of thioredoxin mog fusion protein |
| topic | experimental autoimmune encephalomyelitis multiple sclerosis myelin oligodendrocyte glycoprotein response surface methodology |
| url | https://ps.tbzmed.ac.ir/PDF/ps-30-252.pdf |
| work_keys_str_mv | AT maryamradmard responsesurfacemethodologyapproachtooptimizetheexpressionofthioredoxinmogfusionprotein AT atiehhashemi responsesurfacemethodologyapproachtooptimizetheexpressionofthioredoxinmogfusionprotein |