Glycosylation and Serological Reactivity of an Expression-enhanced SARS-CoV-2 Viral Spike Mimetic.
Antibodies, Viral
/ blood
Binding Sites
/ genetics
COVID-19
/ immunology
Glycosylation
Humans
Immunoglobulin A
/ blood
Immunoglobulin G
/ blood
Immunoglobulin M
/ blood
Mannose
/ metabolism
Mutation, Missense
/ genetics
Oligosaccharides
/ metabolism
Polysaccharides
/ metabolism
Proline
/ genetics
Recombinant Proteins
/ chemistry
SARS-CoV-2
/ immunology
Spike Glycoprotein, Coronavirus
/ genetics
SARS-CoV-2
antibody
glycoprotein
glycosylation
serology
Journal
Journal of molecular biology
ISSN: 1089-8638
Titre abrégé: J Mol Biol
Pays: Netherlands
ID NLM: 2985088R
Informations de publication
Date de publication:
30 01 2022
30 01 2022
Historique:
received:
09
08
2021
revised:
21
10
2021
accepted:
21
10
2021
pubmed:
1
11
2021
medline:
27
1
2022
entrez:
31
10
2021
Statut:
ppublish
Résumé
Extensive glycosylation of viral glycoproteins is a key feature of the antigenic surface of viruses and yet glycan processing can also be influenced by the manner of their recombinant production. The low yields of the soluble form of the trimeric spike (S) glycoprotein from SARS-CoV-2 has prompted advances in protein engineering that have greatly enhanced the stability and yields of the glycoprotein. The latest expression-enhanced version of the spike incorporates six proline substitutions to stabilize the prefusion conformation (termed SARS-CoV-2 S HexaPro). Although the substitutions greatly enhanced expression whilst not compromising protein structure, the influence of these substitutions on glycan processing has not been explored. Here, we show that the site-specific N-linked glycosylation of the expression-enhanced HexaPro resembles that of an earlier version containing two proline substitutions (2P), and that both capture features of native viral glycosylation. However, there are site-specific differences in glycosylation of HexaPro when compared to 2P. Despite these discrepancies, analysis of the serological reactivity of clinical samples from infected individuals confirmed that both HexaPro and 2P protein are equally able to detect IgG, IgA, and IgM responses in all sera analysed. Moreover, we extend this observation to include an analysis of glycan engineered S protein, whereby all N-linked glycans were converted to oligomannose-type and conclude that serological activity is not impacted by large scale changes in glycosylation. These observations suggest that variations in glycan processing will not impact the serological assessments currently being performed across the globe.
Identifiants
pubmed: 34717971
pii: S0022-2836(21)00569-6
doi: 10.1016/j.jmb.2021.167332
pmc: PMC8550889
pii:
doi:
Substances chimiques
Antibodies, Viral
0
Immunoglobulin A
0
Immunoglobulin G
0
Immunoglobulin M
0
Oligosaccharides
0
Polysaccharides
0
Recombinant Proteins
0
Spike Glycoprotein, Coronavirus
0
spike protein, SARS-CoV-2
0
Proline
9DLQ4CIU6V
Mannose
PHA4727WTP
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
167332Subventions
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Bill & Melinda Gates Foundation
ID : INV-008352
Pays : United States
Organisme : Medical Research Council
ID : MR/N023706/1
Pays : United Kingdom
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests J.S.M. is an inventor on the following U.S. patent applications: no. 62/412,703 (“Prefusion Coronavirus Spike Proteins and Their Use”); no. 62/972,886 (“2019-nCoV Vaccine”); no. 63/032,502 (“Engineered Coronavirus Spike (S) Protein and Methods of Use Thereof”).
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