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KMID : 0545120150250081307
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Journal of Microbiology and Biotechnology
2015 Volume.25 No. 8 p.1307 ~ p.1314
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Expression and Efficient One-Step Chromatographic Purification of a Soluble Antagonist for Human Leukemia Inhibitory Factor Receptor in Escherichia coli
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Kim Eun-Yeong
Choi Hee-Jung
Chung Tae-Wook
Jang Se-Bok
Kim Ki-Bong
Ha Ki-Tae
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Abstract
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Leukemia inhibitory factor (LIF) is a member of the IL-6 cytokine family, having pleiotropic actions such as maintaining stem cell pluripotency and enabling blastocyst implantation. Because the action of LIF is mediated by a ligand-receptor interaction with the LIF receptor (LIF-R), an antagonist for LIF-R has been developed to inhibit LIF-induced signaling. In this study, we present a novel method for the production and purification of an antagonist to human LIF-R (hLA). His-tagged hLA was expressed in E. coli, and simple purification methods without any endopeptidase cleavage were designed. In addition, we determined the optimal temperature conditions for enhancing the production of soluble hLA. Finally, the bioactivity of His-tagged hLA was examined using STAT3 phosphorylation and receptivity of human endometrial ECC-1 cells. Our strategy provides a rapid and efficient method to produce biologically active recombinant hLA.
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KEYWORD
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Leukemia inhibitory factor, Escherichia coli, Antagonist, LIF-05, His-tagged, Endometrial cells
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