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http://dx.doi.org/10.4014/jmb.1501.01094

Expression and Efficient One-Step Chromatographic Purification of a Soluble Antagonist for Human Leukemia Inhibitory Factor Receptor in Escherichia coli  

Kim, Eun-Yeong (Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University)
Choi, Hee-Jung (Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University)
Chung, Tae-Wook (Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University)
Jang, Se Bok (Department of Molecular Biology, College of Natural Sciences, Pusan National University)
Kim, Kibong (2nd Division of Clinical Medicine, School of Korean Medicine, Pusan National University and Pediatric, Korean Medicine Hospital, Pusan National University Hospital)
Ha, Ki-Tae (Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.8, 2015 , pp. 1307-1314 More about this Journal
Abstract
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.
Keywords
Leukemia inhibitory factor; Escherichia coli; antagonist; LIF-05; His-tagged; endometrial cells;
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