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Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation
Transient gene expression (TGE) is a methodology employed in bioprocessing for the fast provision of recombinant protein material. Mild hypothermia is often introduced to overcome the low yield typically achieved with TGE and improve specific protein productivity. It is therefore of interest to exam...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5765507/ https://www.ncbi.nlm.nih.gov/pubmed/28921534 http://dx.doi.org/10.1002/bit.26456 |
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author | Sou, Si N. Lee, Ken Nayyar, Kalpana Polizzi, Karen M. Sellick, Christopher Kontoravdi, Cleo |
author_facet | Sou, Si N. Lee, Ken Nayyar, Kalpana Polizzi, Karen M. Sellick, Christopher Kontoravdi, Cleo |
author_sort | Sou, Si N. |
collection | PubMed |
description | Transient gene expression (TGE) is a methodology employed in bioprocessing for the fast provision of recombinant protein material. Mild hypothermia is often introduced to overcome the low yield typically achieved with TGE and improve specific protein productivity. It is therefore of interest to examine the impact of mild hypothermic temperatures on both the yield and quality of transiently expressed proteins and the relationship to changes in cellular processes and metabolism. In this study, we focus on the ability of a Chinese hamster ovary cell line to galactosylate a recombinant monoclonal antibody (mAb) product. Through experimentation and flux balance analysis, our results show that TGE in mild hypothermic conditions led to a 76% increase in q(P) compared to TGE at 36.5°C in our system. This increase is accompanied by increased consumption of nutrients and amino acids, together with increased production of intracellular nucleotide sugar species, and higher rates of mAb galactosylation, despite a reduced rate of cell growth. The reduction in biomass accumulation allowed cells to redistribute their energy and resources toward mAb synthesis and Fc‐glycosylation. Interestingly, the higher capacity of cells to galactosylate the recombinant product in TGE at 32°C appears not to have been assisted by the upregulation of galactosyltransferases (GalTs), but by the increased expression of N‐acetylglucosaminyltransferase II (GnTII) in this cell line, which facilitated the production of bi‐antennary glycan structures for further processing. |
format | Online Article Text |
id | pubmed-5765507 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-57655072018-02-01 Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation Sou, Si N. Lee, Ken Nayyar, Kalpana Polizzi, Karen M. Sellick, Christopher Kontoravdi, Cleo Biotechnol Bioeng COMMUNICATION TO THE EDITORS Transient gene expression (TGE) is a methodology employed in bioprocessing for the fast provision of recombinant protein material. Mild hypothermia is often introduced to overcome the low yield typically achieved with TGE and improve specific protein productivity. It is therefore of interest to examine the impact of mild hypothermic temperatures on both the yield and quality of transiently expressed proteins and the relationship to changes in cellular processes and metabolism. In this study, we focus on the ability of a Chinese hamster ovary cell line to galactosylate a recombinant monoclonal antibody (mAb) product. Through experimentation and flux balance analysis, our results show that TGE in mild hypothermic conditions led to a 76% increase in q(P) compared to TGE at 36.5°C in our system. This increase is accompanied by increased consumption of nutrients and amino acids, together with increased production of intracellular nucleotide sugar species, and higher rates of mAb galactosylation, despite a reduced rate of cell growth. The reduction in biomass accumulation allowed cells to redistribute their energy and resources toward mAb synthesis and Fc‐glycosylation. Interestingly, the higher capacity of cells to galactosylate the recombinant product in TGE at 32°C appears not to have been assisted by the upregulation of galactosyltransferases (GalTs), but by the increased expression of N‐acetylglucosaminyltransferase II (GnTII) in this cell line, which facilitated the production of bi‐antennary glycan structures for further processing. John Wiley and Sons Inc. 2017-11-16 2018-02 /pmc/articles/PMC5765507/ /pubmed/28921534 http://dx.doi.org/10.1002/bit.26456 Text en © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | COMMUNICATION TO THE EDITORS Sou, Si N. Lee, Ken Nayyar, Kalpana Polizzi, Karen M. Sellick, Christopher Kontoravdi, Cleo Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation |
title | Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation |
title_full | Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation |
title_fullStr | Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation |
title_full_unstemmed | Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation |
title_short | Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc‐glycosylation |
title_sort | exploring cellular behavior under transient gene expression and its impact on mab productivity and fc‐glycosylation |
topic | COMMUNICATION TO THE EDITORS |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5765507/ https://www.ncbi.nlm.nih.gov/pubmed/28921534 http://dx.doi.org/10.1002/bit.26456 |
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