Cell-Free Glycoengineering of the Recombinant SARS-CoV-2 Spike Glycoprotein.
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Authors
Ruhnau, JohannesGrote, Valerian
Juarez-Osorio, Mariana
Bruder, Dunja

Mahour, Reza
Rapp, Erdmann
Rexer, Thomas F T
Reichl, Udo
Issue Date
2021-08-16
Metadata
Show full item recordAbstract
The baculovirus-insect cell expression system is readily utilized to produce viral glycoproteins for research as well as for subunit vaccines and vaccine candidates, for instance against SARS-CoV-2 infections. However, the glycoforms of recombinant proteins derived from this expression system are inherently different from mammalian cell-derived glycoforms with mainly complex-type N-glycans attached, and the impact of these differences in protein glycosylation on the immunogenicity is severely under investigated. This applies also to the SARS-CoV-2 spike glycoprotein, which is the antigen target of all licensed vaccines and vaccine candidates including virus like particles and subunit vaccines that are variants of the spike protein. Here, we expressed the transmembrane-deleted human β-1,2 N-acetlyglucosamintransferases I and II (MGAT1ΔTM and MGAT2ΔTM) and the β-1,4-galactosyltransferase (GalTΔTM) in E. coli to in-vitro remodel the N-glycans of a recombinant SARS-CoV-2 spike glycoprotein derived from insect cells. In a cell-free sequential one-pot reaction, fucosylated and afucosylated paucimannose-type N-glycans were converted to complex-type galactosylated N-glycans. In the future, this in-vitro glycoengineering approach can be used to efficiently generate a wide range of N-glycans on antigens considered as vaccine candidates for animal trials and preclinical testing to better characterize the impact of N-glycosylation on immunity and to improve the efficacy of protein subunit vaccines.Citation
Front Bioeng Biotechnol. 2021 Aug 16;9:699025. doi: 10.3389/fbioe.2021.699025.Affiliation
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.Publisher
FrontiersPubMed ID
34485255Type
ArticleLanguage
enISSN
2296-4185ae974a485f413a2113503eed53cd6c53
10.3389/fbioe.2021.699025
Scopus Count
The following license files are associated with this item:
- Creative Commons
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