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Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells

The human embryonal kidney 293 cell (HEK‐293) is a widely used expression host for transient gene expression. The genes or plasmids used for the transient transfections are usually propagated and extracted from the gram‐negative bacterium Escherichia coli, the workhorse for molecular biologists. As...

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Autores principales: Faust, Christine, Beil, Christian, Dittrich, Werner, Rao, Ercole, Langer, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8576075/
https://www.ncbi.nlm.nih.gov/pubmed/34764829
http://dx.doi.org/10.1002/elsc.202100065
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author Faust, Christine
Beil, Christian
Dittrich, Werner
Rao, Ercole
Langer, Thomas
author_facet Faust, Christine
Beil, Christian
Dittrich, Werner
Rao, Ercole
Langer, Thomas
author_sort Faust, Christine
collection PubMed
description The human embryonal kidney 293 cell (HEK‐293) is a widely used expression host for transient gene expression. The genes or plasmids used for the transient transfections are usually propagated and extracted from the gram‐negative bacterium Escherichia coli, the workhorse for molecular biologists. As a gram‐negative bacterium E. coli has an outer membrane (OM) containing lipopolysaccharides (LPS) or endotoxins. LPS are very potent inducers of inflammatory cytokines in the body. In early research phases DNA intended for transient transfections is not routinely checked for LPS‐levels. In this study we addressed the question whether LPS has an impact on the cultivation and production of a recombinant antibody. At high concentrations the presence of LPS has a detrimental impact on cell viability and recombinant protein expression. But low LPS concentrations are tolerated and might even enhance protein expression levels.
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spelling pubmed-85760752021-11-10 Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells Faust, Christine Beil, Christian Dittrich, Werner Rao, Ercole Langer, Thomas Eng Life Sci Research Articles The human embryonal kidney 293 cell (HEK‐293) is a widely used expression host for transient gene expression. The genes or plasmids used for the transient transfections are usually propagated and extracted from the gram‐negative bacterium Escherichia coli, the workhorse for molecular biologists. As a gram‐negative bacterium E. coli has an outer membrane (OM) containing lipopolysaccharides (LPS) or endotoxins. LPS are very potent inducers of inflammatory cytokines in the body. In early research phases DNA intended for transient transfections is not routinely checked for LPS‐levels. In this study we addressed the question whether LPS has an impact on the cultivation and production of a recombinant antibody. At high concentrations the presence of LPS has a detrimental impact on cell viability and recombinant protein expression. But low LPS concentrations are tolerated and might even enhance protein expression levels. John Wiley and Sons Inc. 2021-10-25 /pmc/articles/PMC8576075/ /pubmed/34764829 http://dx.doi.org/10.1002/elsc.202100065 Text en © 2021 The Authors. Engineering in Life Sciences published by Wiley‐VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Faust, Christine
Beil, Christian
Dittrich, Werner
Rao, Ercole
Langer, Thomas
Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
title Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
title_full Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
title_fullStr Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
title_full_unstemmed Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
title_short Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
title_sort impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (hek‐293) cells
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8576075/
https://www.ncbi.nlm.nih.gov/pubmed/34764829
http://dx.doi.org/10.1002/elsc.202100065
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