Assessing Antigen Structural Integrity through Glycosylation Analysis of the SARS-CoV‑2 Viral Spike
journal contributionposted on 08.04.2021, 12:03 by Juliane Brun, Snežana Vasiljevic, Bevin Gangadharan, Mario Hensen, Anu V. Chandran, Michelle L. Hill, J.L. Kiappes, Raymond A. Dwek, Dominic S. Alonzi, Weston B. Struwe, Nicole Zitzmann
Severe acute respiratory syndrome coronavirus 2 is the causative pathogen of the COVID-19 pandemic which as of March 29, 2021, has claimed 2 776 175 lives worldwide. Vaccine development efforts focus on the viral trimeric spike glycoprotein as the main target of the humoral immune response. Viral spikes carry glycans that facilitate immune evasion by shielding specific protein epitopes from antibody neutralization, and antigen efficacy is influenced by spike glycoprotein production in vivo. Therefore, immunogen integrity is important for glycoprotein-based vaccine candidates. Here, we show how site-specific glycosylation differs between virus-derived spikes, wild-type, non-stabilized spikes expressed from a plasmid with a CMV promoter and tPA signal sequence, and commonly used recombinant, engineered spike glycoproteins. Furthermore, we show that their distinctive cellular secretion pathways result in different protein glycosylation and secretion patterns, including shedding of spike monomeric subunits for the non-stabilized wild-type spike tested, which may have implications for the resulting immune response and vaccine design.
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2 776 175 livesantigen efficacynon-stabilized wild-type spikeCMV promotersyndrome coronavirus 2antibody neutralizationCOVID -19 pandemicGlycosylation AnalysisVaccine development efforts focussite-specific glycosylationViral spikesnon-stabilized spikesprotein glycosylationsecretion patternstrimeric spike glycoproteinglycoprotein-based vaccine candidatesspike glycoproteins2 776 175spike glycoprotein productionvirus-derived spikestPA signal sequenceAssessing Antigen Structural Integrityvaccine designsecretion pathways resultimmunogen integrityprotein epitopesspike monomeric subunits