Park, H Im, H Kang, YJ Yu, MH Hong, HJ 2024-01-21T13:31:52Z 2024-01-21T13:31:52Z 2022-01-10 2000-10 0141-5492 https://pubs.kist.re.kr/handle/201004/141059 The extracellular domain (edMpl) of human thrombopoietin (TPO) receptor, c-Mpl was expressed in Escherichia coli by changing some nucleotides before and after the translation initiation codon. The mutations increased the expression by approx. 15-fold. The inclusion bodies were solubilized in 8 M guanidine-HCl under reducing conditions and refolded using a glutathione-redox system. The monomeric form of edMpl was purified to near homogeneity by two successive steps of ion-exchange chromatography using DEAE-Sephacel and Mono Q columns. The purified monomeric edMpl inhibited the TPO-dependent cell proliferation, suggesting that it was binding to TPO. Also, antisera raised against the edMpl bound specifically to the soluble receptor secreted by mammalian cells. English KLUWER ACADEMIC PUBL C-MPL LIGAND IDENTIFICATION CLONING STIMULATION GROWTH CODON Expression, purification and characterization of soluble human thrombopoietin receptor from Escherichia coli Article 10.1023/A:1005672824663 1 BIOTECHNOLOGY LETTERS, v.22, no.20, pp.1611 - 1617 BIOTECHNOLOGY LETTERS 22 20 1611 1617 scie scopus 000089975000006 Biotechnology & Applied Microbiology Biotechnology & Applied Microbiology Article C-MPL LIGAND IDENTIFICATION CLONING STIMULATION GROWTH CODON inclusion bodies protein purification receptor domain thrombopoietin