TY  - JOUR
A1  - Zemella, Anne
A1  - Thoring, Lena
A1  - Hoffmeister, Christian
A1  - Samalikova, Maria
A1  - Ehren, Patricia
A1  - Wüstenhagen, Doreen Anja
A1  - Kubick, Stefan
T1  - Cell-free protein synthesis as a novel tool for directed glycoengineering of active erythropoietin
JF  - Scientific reports
N2  - 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-directed incorporation of non-canonical amino acids with subsequent chemoselective modifications.
Y1  - 2018
U6  - https://doi.org/10.1038/s41598-018-26936-x
SN  - 2045-2322
VL  - 8
PB  - Nature Publ. Group
CY  - London
ER  - 
TY  - GEN
A1  - Zemella, Anne
A1  - Thoring, Lena
A1  - Hoffmeister, Christian
A1  - Šamalíková, Mária
A1  - Ehren, Patricia
A1  - Wüstenhagen, Doreen Anja
A1  - Kubick, Stefan
T1  - Cell-free protein synthesis as a novel tool for directed glycoengineering of active erythropoietin
T2  - Postprints der Universität Potsdam : Mathematisch Naturwissenschaftliche Reihe
N2  - 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-directed incorporation of non-canonical amino acids with subsequent chemoselective modifications.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 824 
KW  - recombinat-human-erythropoietin
KW  - glycosylation
KW  - expression
KW  - site
KW  - anemia
KW  - CDNA
KW  - glycoprotein
KW  - purification
KW  - cloning
KW  - growth
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-427017
IS  - 824
ER  -