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[Preprint]. 2021 Mar 9:2021.03.08.433764. [Version 1] doi: 10.1101/2021.03.08.433764

Site-specific steric control of SARS-CoV-2 spike glycosylation

Joel D Allen, Himanshi Chawla, Firdaus Samsudin, Lorena Zuzic, Aishwary Tukaram Shivgan, Yasunori Watanabe, Wan-ting He, Sean Callaghan, Ge Song, Peter Yong, Philip J M Brouwer, Yutong Song, Yongfei Cai, Helen M E Duyvesteyn, Tomas Malinauskas, Joeri Kint, Paco Pino, Maria J Wurm, Martin Frank, Bing Chen, David I Stuart, Rogier W Sanders, Raiees Andrabi, Dennis R Burton, Sai Li, Peter J Bond, Max Crispin
PMCID: PMC7986994  PMID: 33758835

Abstract

A central tenet in the design of vaccines is the display of native-like antigens in the elicitation of protective immunity. The abundance of N-linked glycans across the SARS-CoV-2 spike protein is a potential source of heterogeneity between the many different vaccine candidates under investigation. Here, we investigate the glycosylation of recombinant SARS-CoV-2 spike proteins from five different laboratories and compare them against infectious virus S protein. We find patterns which are conserved across all samples and this can be associated with site-specific stalling of glycan maturation which act as a highly sensitive reporter of protein structure. Molecular dynamics (MD) simulations of a fully glycosylated spike support s a model of steric restrictions that shape enzymatic processing of the glycans. These results suggest that recombinant spike-based SARS-CoV-2 immunogen glycosylation reproducibly recapitulates signatures of viral glycosylation.

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