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Cell-free protein synthesis as a novel tool for directed glycoengineering of active erythropoietin
- As one of the most complex post-translational modification, glycosylation is widely involved in cell adhesion, cell proliferation and immune response. Nevertheless glycoproteins with an identical polypeptide backbone mostly differ in their glycosylation patterns. Due to this heterogeneity, the mapping of different glycosylation patterns to their associated function is nearly impossible. In the last years, glycoengineering tools including cell line engineering, chemoenzymatic remodeling and site-specific glycosylation have attracted increasing interest. The therapeutic hormone erythropoietin (EPO) has been investigated in particular by various groups to establish a production process resulting in a defined glycosylation pattern. However commercially available recombinant human EPO shows batch-to-batch variations in its glycoforms. Therefore we present an alternative method for the synthesis of active glycosylated EPO with an engineered O-glycosylation site by combining eukaryotic cell-free protein synthesis and site-directedAs one of the most complex post-translational modification, glycosylation is widely involved in cell adhesion, cell proliferation and immune response. Nevertheless glycoproteins with an identical polypeptide backbone mostly differ in their glycosylation patterns. Due to this heterogeneity, the mapping of different glycosylation patterns to their associated function is nearly impossible. In the last years, glycoengineering tools including cell line engineering, chemoenzymatic remodeling and site-specific glycosylation have attracted increasing interest. The therapeutic hormone erythropoietin (EPO) has been investigated in particular by various groups to establish a production process resulting in a defined glycosylation pattern. However commercially available recombinant human EPO shows batch-to-batch variations in its glycoforms. Therefore we present an alternative method for the synthesis of active glycosylated EPO with an engineered O-glycosylation site by combining eukaryotic cell-free protein synthesis and site-directed incorporation of non-canonical amino acids with subsequent chemoselective modifications.…
Author details: | Anne ZemellaGND, Lena ThoringGND, Christian Hoffmeister, Mária Šamalíková, Patricia Ehren, Doreen Anja Wüstenhagen, Stefan KubickORCiD |
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URN: | urn:nbn:de:kobv:517-opus4-427017 |
DOI: | https://doi.org/10.25932/publishup-42701 |
Title of parent work (German): | Postprints der Universität Potsdam : Mathematisch Naturwissenschaftliche Reihe |
Publication series (Volume number): | Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe (824) |
Publication type: | Postprint |
Language: | English |
Date of first publication: | 2020/02/27 |
Publication year: | 2018 |
Publishing institution: | Universität Potsdam |
Release date: | 2020/02/27 |
Tag: | CDNA; anemia; cloning; expression; glycoprotein; glycosylation; growth; purification; recombinat-human-erythropoietin; site |
Issue: | 824 |
Number of pages: | 14 |
Source: | Scientific Reports 8 (2018) 8514 DOI:10.1038/s41598-018-26936-x |
Organizational units: | Mathematisch-Naturwissenschaftliche Fakultät |
DDC classification: | 5 Naturwissenschaften und Mathematik / 50 Naturwissenschaften / 500 Naturwissenschaften und Mathematik |
6 Technik, Medizin, angewandte Wissenschaften / 60 Technik / 600 Technik, Technologie | |
Peer review: | Referiert |
Publishing method: | Open Access |
License (German): | CC-BY - Namensnennung 4.0 International |
External remark: | Bibliographieeintrag der Originalveröffentlichung/Quelle |