Gieseler, G.-M.; Ekramzadeh, K.; Nölle, V.; Malysheva, S.; Kempf, H. et al.: Solubilization and renaturation of biologically active human bone morphogenetic protein-4 from inclusion bodies. In: Biotechnology Reports 18 (2018), Nr., e00249. DOI: https://doi.org/10.1016/j.btre.2018.e00249
Abstract: | |
Biologically active human bone morphogenetic protein-4 (hBMP-4) was successfully produced in a prokaryotic host. For this aim, hBMP-4 cDNA was cloned in Escherichia coli (E. coli) and the protein was produced in a non-active aggregated form. After washing and solubilization, in vitro refolding of the rhBMP-4 monomer was performed using rapid dilution. In this study, different refolding conditions were tested for the dimerization of rhBMP-4 by one-factor-at-a-time variation. The dimerization process was found to be sensitive to pH, protein concentration and the presence of aggregation suppressors. In contrast, redox conditions and ionic strength did not impact refolding as expected. The dimer was separated from the remaining monomer, aggregates and host cell contaminants in a single step using cation-exchange membrane chromatography. The rhBMP-4 dimer produced in E. coli was biologically active as demonstrated by its capability to induce trophoblast differentiation and primitive streak induction of human pluripotent stem cells (hPSCs). © 2018 | |
License of this version: | CC BY 4.0 Unported |
Document Type: | Article |
Publishing status: | publishedVersion |
Issue Date: | 2018 |
Appears in Collections: | Naturwissenschaftliche Fakultät |
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