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.…
Verfasserangaben: | Anne ZemellaGND, Lena ThoringGND, Christian Hoffmeister, Maria Samalikova, Patricia Ehren, Doreen Anja Wüstenhagen, Stefan KubickORCiD |
---|---|
DOI: | https://doi.org/10.1038/s41598-018-26936-x |
ISSN: | 2045-2322 |
Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/29867209 |
Titel des übergeordneten Werks (Englisch): | Scientific reports |
Verlag: | Nature Publ. Group |
Verlagsort: | London |
Publikationstyp: | Wissenschaftlicher Artikel |
Sprache: | Englisch |
Datum der Erstveröffentlichung: | 04.06.2018 |
Erscheinungsjahr: | 2018 |
Datum der Freischaltung: | 22.11.2021 |
Band: | 8 |
Seitenanzahl: | 12 |
Fördernde Institution: | European Regional Development Fund (EFRE)European Union (EU); German Ministry of Education and Research (BMBF)Federal Ministry of Education & Research (BMBF) [031B0078A]; German Research Foundation (DFG)German Research Foundation (DFG) [1623] |
Organisationseinheiten: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Mathematik |
DDC-Klassifikation: | 5 Naturwissenschaften und Mathematik / 50 Naturwissenschaften / 500 Naturwissenschaften und Mathematik |
6 Technik, Medizin, angewandte Wissenschaften / 60 Technik / 600 Technik, Technologie | |
Peer Review: | Referiert |
Publikationsweg: | Open Access / Gold Open-Access |
DOAJ gelistet | |
Lizenz (Deutsch): | CC-BY - Namensnennung 4.0 International |
Externe Anmerkung: | Zweitveröffentlichung in der Schriftenreihe Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe ; 824 |