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

Abstract 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 biolog...

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Autores principales:	Christine Faust, Christian Beil, Werner Dittrich, Ercole Rao, Thomas Langer
Formato:	article
Lenguaje:	EN
Publicado:	Wiley-VCH 2021
Materias:	endotoxin HEK‐293 cells lipopolysaccharides recombinant protein expression transient transfection Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/e7d73704dc174b76b633edeb7e530783
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spelling	oai:doaj.org-article:e7d73704dc174b76b633edeb7e5307832021-11-09T05:07:06ZImpact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells1618-28631618-024010.1002/elsc.202100065https://doaj.org/article/e7d73704dc174b76b633edeb7e5307832021-11-01T00:00:00Zhttps://doi.org/10.1002/elsc.202100065https://doaj.org/toc/1618-0240https://doaj.org/toc/1618-2863Abstract 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.Christine FaustChristian BeilWerner DittrichErcole RaoThomas LangerWiley-VCHarticleendotoxinHEK‐293 cellslipopolysaccharidesrecombinant protein expressiontransient transfectionBiotechnologyTP248.13-248.65ENEngineering in Life Sciences, Vol 21, Iss 11, Pp 778-785 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	endotoxin HEK‐293 cells lipopolysaccharides recombinant protein expression transient transfection Biotechnology TP248.13-248.65
spellingShingle	endotoxin HEK‐293 cells lipopolysaccharides recombinant protein expression transient transfection Biotechnology TP248.13-248.65 Christine Faust Christian Beil Werner Dittrich Ercole Rao Thomas Langer Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
description	Abstract 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.
format	article
author	Christine Faust Christian Beil Werner Dittrich Ercole Rao Thomas Langer
author_facet	Christine Faust Christian Beil Werner Dittrich Ercole Rao Thomas Langer
author_sort	Christine Faust
title	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_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_sort	impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (hek‐293) cells
publisher	Wiley-VCH
publishDate	2021
url	https://doaj.org/article/e7d73704dc174b76b633edeb7e530783
work_keys_str_mv	AT christinefaust impactoflipopolysaccharidesoncultivationandrecombinantproteinexpressioninhumanembryonalkidneyhek293cells AT christianbeil impactoflipopolysaccharidesoncultivationandrecombinantproteinexpressioninhumanembryonalkidneyhek293cells AT wernerdittrich impactoflipopolysaccharidesoncultivationandrecombinantproteinexpressioninhumanembryonalkidneyhek293cells AT ercolerao impactoflipopolysaccharidesoncultivationandrecombinantproteinexpressioninhumanembryonalkidneyhek293cells AT thomaslanger impactoflipopolysaccharidesoncultivationandrecombinantproteinexpressioninhumanembryonalkidneyhek293cells
_version_	1718441344806944768

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

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