Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells
Abstract Interferon-β-1a (INF-β-1a) has gained significant attention due to its emerging applications in the treatment of different human diseases. Therefore, many researchers have attempted to produce it in large quantities and also in a biologically active form using different expression systems....
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Universidade de São Paulo
2022-03-01
|
| Series: | Brazilian Journal of Pharmaceutical Sciences |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502022000100528&tlng=en |
| _version_ | 1811273528119394304 |
|---|---|
| author | Nasim Moatamedi Rahman Emamzadeh Hamid Mir Mohammad Sadeghi Vajihe Akbari |
| author_facet | Nasim Moatamedi Rahman Emamzadeh Hamid Mir Mohammad Sadeghi Vajihe Akbari |
| author_sort | Nasim Moatamedi |
| collection | DOAJ |
| description | Abstract Interferon-β-1a (INF-β-1a) has gained significant attention due to its emerging applications in the treatment of different human diseases. Therefore, many researchers have attempted to produce it in large quantities and also in a biologically active form using different expression systems. In the present study, we aimed to improve the expression level of INF-β-1a by Pichia pastoris using optimization of culture conditions. The codon-optimized INF-β- 1a gene was cloned into pPICZαA plasmid under the control of alcohol oxidase I (AOX1) promoter. The protein expression was induced using different concentrations of methanol at different pHs and temperatures. The biological activity of produced protein was evaluated by anti-proliferative assay. The ideal culture conditions for the expression of INF-β-1a by P. pastoris were found to be induction with 2% methanol at pH 7.0 culture medium at 30 C which yielded a concentration of 15.5 mg/L INF-β-1a in a shake flask. Our results indicate that differences in glycosylation pattern could result in different biological activities as INF- β-1a produced by P. pastoris could significantly more reduce the cell viability of HepG-2 cells, a hepatocellular carcinoma cell line, than a commercially available form of this protein produced by CHO. |
| first_indexed | 2024-04-12T23:00:49Z |
| format | Article |
| id | doaj.art-06d4894154f24f1fa375aa2d1b00dd02 |
| institution | Directory Open Access Journal |
| issn | 2175-9790 |
| language | English |
| last_indexed | 2024-04-12T23:00:49Z |
| publishDate | 2022-03-01 |
| publisher | Universidade de São Paulo |
| record_format | Article |
| series | Brazilian Journal of Pharmaceutical Sciences |
| spelling | doaj.art-06d4894154f24f1fa375aa2d1b00dd022022-12-22T03:13:03ZengUniversidade de São PauloBrazilian Journal of Pharmaceutical Sciences2175-97902022-03-015810.1590/s2175-97902022e18984Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cellsNasim Moatamedihttps://orcid.org/0000-0002-4289-110XRahman Emamzadehhttps://orcid.org/0000-0002-4144-4332Hamid Mir Mohammad Sadeghihttps://orcid.org/0000-0002-4834-4502Vajihe Akbarihttps://orcid.org/0000-0003-0754-1486Abstract Interferon-β-1a (INF-β-1a) has gained significant attention due to its emerging applications in the treatment of different human diseases. Therefore, many researchers have attempted to produce it in large quantities and also in a biologically active form using different expression systems. In the present study, we aimed to improve the expression level of INF-β-1a by Pichia pastoris using optimization of culture conditions. The codon-optimized INF-β- 1a gene was cloned into pPICZαA plasmid under the control of alcohol oxidase I (AOX1) promoter. The protein expression was induced using different concentrations of methanol at different pHs and temperatures. The biological activity of produced protein was evaluated by anti-proliferative assay. The ideal culture conditions for the expression of INF-β-1a by P. pastoris were found to be induction with 2% methanol at pH 7.0 culture medium at 30 C which yielded a concentration of 15.5 mg/L INF-β-1a in a shake flask. Our results indicate that differences in glycosylation pattern could result in different biological activities as INF- β-1a produced by P. pastoris could significantly more reduce the cell viability of HepG-2 cells, a hepatocellular carcinoma cell line, than a commercially available form of this protein produced by CHO.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502022000100528&tlng=enInterferon-β-1aPichia pastorisExpressionCell survivalHepatocellular carcinoma |
| spellingShingle | Nasim Moatamedi Rahman Emamzadeh Hamid Mir Mohammad Sadeghi Vajihe Akbari Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells Brazilian Journal of Pharmaceutical Sciences Interferon-β-1a Pichia pastoris Expression Cell survival Hepatocellular carcinoma |
| title | Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells |
| title_full | Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells |
| title_fullStr | Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells |
| title_full_unstemmed | Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells |
| title_short | Bioprocess optimization of interferon β-1-a in Pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells |
| title_sort | bioprocess optimization of interferon β 1 a in pichia pastoris and its improved inhibitory effect against hepatocellular carcinoma cells |
| topic | Interferon-β-1a Pichia pastoris Expression Cell survival Hepatocellular carcinoma |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502022000100528&tlng=en |
| work_keys_str_mv | AT nasimmoatamedi bioprocessoptimizationofinterferonb1ainpichiapastorisanditsimprovedinhibitoryeffectagainsthepatocellularcarcinomacells AT rahmanemamzadeh bioprocessoptimizationofinterferonb1ainpichiapastorisanditsimprovedinhibitoryeffectagainsthepatocellularcarcinomacells AT hamidmirmohammadsadeghi bioprocessoptimizationofinterferonb1ainpichiapastorisanditsimprovedinhibitoryeffectagainsthepatocellularcarcinomacells AT vajiheakbari bioprocessoptimizationofinterferonb1ainpichiapastorisanditsimprovedinhibitoryeffectagainsthepatocellularcarcinomacells |