Abstract
Leukemia inhibitory factor (LIF) is a polyfunctional cytokine with numerous regulatory effects in vivo and in vitro. In murine stem cell cultures it is the essential media supplement for the maintenance of pluripotency of embryonic and induced pluripotent stem cells. To explore if the glycosylation and/or other post-translational modifications are affecting this activity, we produced and isolated LIF from either eucaryotic cells (Chinese hamster ovary (CHO) cells) or procaryotes (E.coli) and compared their biological activities in this study.
Highlights
Leukemia inhibitory factor (LIF) is a polyfunctional cytokine with numerous regulatory effects in vivo and in vitro
The genes for the fusion protein are separated on the vector by a TEV (Tobacco Etch Virus) protease cleavage site, in addition thioredoxin is expressed with a his-tag
The protein was afterwards purified from the fermentation broth by metal chelate affinity chromatography through the utilization of Zn2+ ions immobilized on IDA membrane adsorbers
Summary
Leukemia inhibitory factor (LIF) is a polyfunctional cytokine with numerous regulatory effects in vivo and in vitro. Production in E.coli To increase the solubility the LIF is expressed together with thioredoxin as a fusion protein. The protein was afterwards purified from the fermentation broth by metal chelate affinity chromatography through the utilization of Zn2+ ions immobilized on IDA membrane adsorbers.
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