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...

Full description

Saved in:
Bibliographic Details
Published in:Engineering in life sciences 2021-11, Vol.21 (11), p.778-785
Main Authors: Faust, Christine, Beil, Christian, Dittrich, Werner, Rao, Ercole, Langer, Thomas
Format: Article
Language:English
Subjects:
Antibodies
Antigens
Bacteria
Cell culture
Cell viability
Cloning
Coliforms
Cultivation
Cytokines
Cytomegalovirus
E coli
endotoxin
Endotoxins
Gene expression
Gram-negative bacteria
HEK‐293 cells
Inflammation
Kidneys
Lipids
Lipopolysaccharides
Monoclonal antibodies
Peptides
Plasmids
Protein expression
Proteins
recombinant protein expression
transient transfection
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1618-0240
1618-2863
DOI:10.1002/elsc.202100065