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The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells
Cell lines derived from mammalian are dominant systems for the production of recombinant therapeutic proteins because of their capacity for correct protein folding, assembly and post-translational modification. In the search of an efficient method for the production of a recombinant protein using an...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Medknow Publications & Media Pvt Ltd
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3764673/ https://www.ncbi.nlm.nih.gov/pubmed/24019831 |
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author | Rezaei, M. Zarkesh-Esfahani, S. H. Gharagozloo, M. |
author_facet | Rezaei, M. Zarkesh-Esfahani, S. H. Gharagozloo, M. |
author_sort | Rezaei, M. |
collection | PubMed |
description | Cell lines derived from mammalian are dominant systems for the production of recombinant therapeutic proteins because of their capacity for correct protein folding, assembly and post-translational modification. In the search of an efficient method for the production of a recombinant protein using animal cell culture, we investigated the effects of different treatment including fetal calf serum concentration, glycerol and culture temperature on a Chinese hamster ovary (CHO) cell line on the production of recombinant human growth hormone (rhGH) and recombinant Chinese hamster ovary (rCHO) viability. The GH production was assessed using ELISA and western blotting methods and cell viability was determined by flow cytometry. The production of recombinant protein increased by 2-fold when stimulatory chemical such as glycerol was added in two stages, first cells were cultured without glycerol for a period of time in order to obtain enough cell density and then glycerol was added to achieve high specific productivity.. Moreover, glycerol addition increased cell viability. Low culture temperature (below 37°C) led to enhanced cellular productivity of the rhGH by 3-fold but decreased cell viability. These findings indicate that quite simple factors such as culture temperature and addition of simple chemicals may lead to the improvement of industrial process for the production of recombinant proteins such as rhGH. |
format | Online Article Text |
id | pubmed-3764673 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Medknow Publications & Media Pvt Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-37646732013-09-09 The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells Rezaei, M. Zarkesh-Esfahani, S. H. Gharagozloo, M. Res Pharm Sci Original Article Cell lines derived from mammalian are dominant systems for the production of recombinant therapeutic proteins because of their capacity for correct protein folding, assembly and post-translational modification. In the search of an efficient method for the production of a recombinant protein using animal cell culture, we investigated the effects of different treatment including fetal calf serum concentration, glycerol and culture temperature on a Chinese hamster ovary (CHO) cell line on the production of recombinant human growth hormone (rhGH) and recombinant Chinese hamster ovary (rCHO) viability. The GH production was assessed using ELISA and western blotting methods and cell viability was determined by flow cytometry. The production of recombinant protein increased by 2-fold when stimulatory chemical such as glycerol was added in two stages, first cells were cultured without glycerol for a period of time in order to obtain enough cell density and then glycerol was added to achieve high specific productivity.. Moreover, glycerol addition increased cell viability. Low culture temperature (below 37°C) led to enhanced cellular productivity of the rhGH by 3-fold but decreased cell viability. These findings indicate that quite simple factors such as culture temperature and addition of simple chemicals may lead to the improvement of industrial process for the production of recombinant proteins such as rhGH. Medknow Publications & Media Pvt Ltd 2013 /pmc/articles/PMC3764673/ /pubmed/24019831 Text en Copyright: © Journal of Research in Pharmaceutical Sciences http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Rezaei, M. Zarkesh-Esfahani, S. H. Gharagozloo, M. The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells |
title | The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells |
title_full | The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells |
title_fullStr | The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells |
title_full_unstemmed | The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells |
title_short | The effect of different media composition and temperatures on the production of recombinant human growth hormone by CHO cells |
title_sort | effect of different media composition and temperatures on the production of recombinant human growth hormone by cho cells |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3764673/ https://www.ncbi.nlm.nih.gov/pubmed/24019831 |
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