Open AccessImpact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK‐293) cells
Author(s) -
Faust Christine,
Beil Christian,
Dittrich Werner,
Rao Ercole,
Langer Thomas
Publication year - 2021
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/elsc.202100065
Subject(s) - recombinant dna , hek 293 cells , escherichia coli , plasmid , biology , bacteria , gene , microbiology and biotechnology , biochemistry , genetics
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.