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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| Veröffentlicht in: | Engineering in life sciences 2021-11, Vol.21 (11), p.778-785 |
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| creator | Faust, Christine Beil, Christian Dittrich, Werner Rao, Ercole Langer, Thomas |
| 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. |
| doi_str_mv | 10.1002/elsc.202100065 |
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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.</description><identifier>ISSN: 1618-0240</identifier><identifier>EISSN: 1618-2863</identifier><identifier>DOI: 10.1002/elsc.202100065</identifier><identifier>PMID: 34764829</identifier><language>eng</language><publisher>HOBOKEN: Wiley</publisher><subject>Antibodies ; Antigens ; Bacteria ; Biotechnology & Applied Microbiology ; Cell culture ; Cell viability ; Cloning ; Coliforms ; Cultivation ; Cytokines ; Cytomegalovirus ; E coli ; endotoxin ; Endotoxins ; Gene expression ; Gram-negative bacteria ; HEK‐293 cells ; Inflammation ; Kidneys ; Life Sciences & Biomedicine ; Lipids ; Lipopolysaccharides ; Monoclonal antibodies ; Peptides ; Plasmids ; Protein expression ; Proteins ; recombinant protein expression ; Science & Technology ; transient transfection</subject><ispartof>Engineering in life sciences, 2021-11, Vol.21 (11), p.778-785</ispartof><rights>2021 The Authors. published by Wiley‐VCH GmbH</rights><rights>2021 The Authors. 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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.</description><subject>Antibodies</subject><subject>Antigens</subject><subject>Bacteria</subject><subject>Biotechnology & Applied Microbiology</subject><subject>Cell culture</subject><subject>Cell viability</subject><subject>Cloning</subject><subject>Coliforms</subject><subject>Cultivation</subject><subject>Cytokines</subject><subject>Cytomegalovirus</subject><subject>E coli</subject><subject>endotoxin</subject><subject>Endotoxins</subject><subject>Gene expression</subject><subject>Gram-negative bacteria</subject><subject>HEK‐293 cells</subject><subject>Inflammation</subject><subject>Kidneys</subject><subject>Life Sciences & Biomedicine</subject><subject>Lipids</subject><subject>Lipopolysaccharides</subject><subject>Monoclonal antibodies</subject><subject>Peptides</subject><subject>Plasmids</subject><subject>Protein expression</subject><subject>Proteins</subject><subject>recombinant protein expression</subject><subject>Science & Technology</subject><subject>transient transfection</subject><issn>1618-0240</issn><issn>1618-2863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>HGBXW</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNkk2P0zAQhiMEYpeFK0cUicsi1OKv2M4FCVWFrajEAThbjj3duiR2sJOFigs_gd_IL8H9oGK5wMnj8TOvZ0ZvUTzGaIoRIi-gTWZKEMkXxKs7xTnmWE6I5PTuMUaEobPiQUobhLCQEt8vzigTnElSnxffFl2vzVCGVdm6PvSh3SZtzFpHZyGVwZdmbAd3oweXY-1tGcGErnFe-6HsYxjA-RK-9hFS2iH5th47nXNdE7fB67b85KyHbXl5NX_78_sPUtNnpYG2TQ-LeyvdJnh0PC-Kj6_nH2ZXk-W7N4vZq-XEVKRGE1FzniOCuLUWamJqwVbMCMwMooZTaIRccVNz1DBsK2JIxQxUGjcNkhUHelEsDro26I3qo-t03KqgndonQrxWOg7OtKBAWEEFYtZgwRrBG04p2PwDVBQJSbPWy4NWPzYdWAN-iLq9JXr7xbu1ug43SlaCI1FlgcujQAyfR0iD6lzarUN7CGNSpMrjScqZyOjTv9BNGGPe6J5ispa83glOD5SJIaUIq1MzGKmdR9TOI-rkkVzw5M8RTvhvU2RAHoAv0IRVMg68gROWRQTOkyCSI4Rnbth7YxZGP-TS5_9fmml2pF0L23_0rebL9zPMGKK_AFd-6lQ</recordid><startdate>202111</startdate><enddate>202111</enddate><creator>Faust, Christine</creator><creator>Beil, Christian</creator><creator>Dittrich, Werner</creator><creator>Rao, Ercole</creator><creator>Langer, Thomas</creator><general>Wiley</general><general>John Wiley & Sons, Inc</general><general>John Wiley and Sons Inc</general><general>Wiley-VCH</general><scope>24P</scope><scope>WIN</scope><scope>BLEPL</scope><scope>DTL</scope><scope>HGBXW</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>L6V</scope><scope>LK8</scope><scope>M7P</scope><scope>M7S</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-4012-3051</orcidid></search><sort><creationdate>202111</creationdate><title>Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells</title><author>Faust, Christine ; 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| subjects | Antibodies Antigens Bacteria Biotechnology & Applied Microbiology Cell culture Cell viability Cloning Coliforms Cultivation Cytokines Cytomegalovirus E coli endotoxin Endotoxins Gene expression Gram-negative bacteria HEK‐293 cells Inflammation Kidneys Life Sciences & Biomedicine Lipids Lipopolysaccharides Monoclonal antibodies Peptides Plasmids Protein expression Proteins recombinant protein expression Science & Technology transient transfection |
| title | Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells |
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