期刊
ENGINEERING IN LIFE SCIENCES
卷 21, 期 11, 页码 778-785出版社
WILEY
DOI: 10.1002/elsc.202100065
关键词
endotoxin; HEK-293 cells; lipopolysaccharides; recombinant protein expression; transient transfection
The presence of LPS from E. coli has a detrimental impact on cell viability and recombinant protein expression at high concentrations, but low concentrations of LPS can be tolerated and even enhance protein expression levels.
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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