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Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells
BACKGROUND: As the demand for monoclonal antibodies (mAb) increases, more efficient expression methods are required for their manufacturing process. Transcriptional gene silencing is a common phenomenon in recombinant cell lines which leads to expression reduction and instability. There are reports...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5322649/ https://www.ncbi.nlm.nih.gov/pubmed/28228095 http://dx.doi.org/10.1186/s12896-017-0330-0 |
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author | Nematpour, Fatemeh Mahboudi, Fereidoun Vaziri, Behrouz Khalaj, Vahid Ahmadi, Samira Ahmadi, Maryam Ebadat, Saedeh Davami, Fatemeh |
author_facet | Nematpour, Fatemeh Mahboudi, Fereidoun Vaziri, Behrouz Khalaj, Vahid Ahmadi, Samira Ahmadi, Maryam Ebadat, Saedeh Davami, Fatemeh |
author_sort | Nematpour, Fatemeh |
collection | PubMed |
description | BACKGROUND: As the demand for monoclonal antibodies (mAb) increases, more efficient expression methods are required for their manufacturing process. Transcriptional gene silencing is a common phenomenon in recombinant cell lines which leads to expression reduction and instability. There are reports on improved antibody expression in ubiquitous chromatin opening element (UCOE) containing both heavy and light chain gene constructs. Here we investigate the impact of having these elements as part of the light chain, heavy chain or both genes during cell line development. In this regard, non-UCOE and UCOE vectors were constructed and stable Chinese hamster ovary (CHO) cell pools were generated by different vector combinations. RESULTS: Expression analysis revealed that all UCOE cell pools had higher antibody yields compared to non-UCOE cells, Moreover the most optimal expression was obtained by cells containing just the UCOE on heavy chain. In terms of stability, it was shown that the high level of expression was kept consistence for more than four months in these cells whereas the expression titers were reduced in the other UCOE pools. CONCLUSIONS: In conclusion, UCOE significantly enhanced the level and stability of antibody expression and the use of this element with heavy chain provided more stable cell lines with higher production level. |
format | Online Article Text |
id | pubmed-5322649 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-53226492017-03-01 Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells Nematpour, Fatemeh Mahboudi, Fereidoun Vaziri, Behrouz Khalaj, Vahid Ahmadi, Samira Ahmadi, Maryam Ebadat, Saedeh Davami, Fatemeh BMC Biotechnol Research Article BACKGROUND: As the demand for monoclonal antibodies (mAb) increases, more efficient expression methods are required for their manufacturing process. Transcriptional gene silencing is a common phenomenon in recombinant cell lines which leads to expression reduction and instability. There are reports on improved antibody expression in ubiquitous chromatin opening element (UCOE) containing both heavy and light chain gene constructs. Here we investigate the impact of having these elements as part of the light chain, heavy chain or both genes during cell line development. In this regard, non-UCOE and UCOE vectors were constructed and stable Chinese hamster ovary (CHO) cell pools were generated by different vector combinations. RESULTS: Expression analysis revealed that all UCOE cell pools had higher antibody yields compared to non-UCOE cells, Moreover the most optimal expression was obtained by cells containing just the UCOE on heavy chain. In terms of stability, it was shown that the high level of expression was kept consistence for more than four months in these cells whereas the expression titers were reduced in the other UCOE pools. CONCLUSIONS: In conclusion, UCOE significantly enhanced the level and stability of antibody expression and the use of this element with heavy chain provided more stable cell lines with higher production level. BioMed Central 2017-02-22 /pmc/articles/PMC5322649/ /pubmed/28228095 http://dx.doi.org/10.1186/s12896-017-0330-0 Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Nematpour, Fatemeh Mahboudi, Fereidoun Vaziri, Behrouz Khalaj, Vahid Ahmadi, Samira Ahmadi, Maryam Ebadat, Saedeh Davami, Fatemeh Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells |
title | Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells |
title_full | Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells |
title_fullStr | Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells |
title_full_unstemmed | Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells |
title_short | Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells |
title_sort | evaluating the expression profile and stability of different ucoe containing vector combinations in mab-producing cho cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5322649/ https://www.ncbi.nlm.nih.gov/pubmed/28228095 http://dx.doi.org/10.1186/s12896-017-0330-0 |
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