Characterization of Site-Specific N- and O-Glycopeptides from Recombinant Spike and ACE2 Glycoproteins Using LC-MS/MS Analysis DOI Open Access

Ju Hwan Song,

S. Y. Jang,

Jinwoong Choi

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(24), P. 13649 - 13649

Published: Dec. 20, 2024

The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in hundreds of millions infections and deaths globally. Although vaccination campaigns are mitigating the emerging viral variants continue to pose challenges. spike (S) protein SARS-CoV-2 plays a critical role entry binding angiotensin-converting enzyme (ACE2) receptor, making both proteins essential targets for therapeutic vaccine development. glycosylation these influences their structure function. This underscores need detailed site-specific glycoproteomic analysis. In this study, we characterized N- or O-glycosylation profiles recombinant receptor-binding domain (RBD) ACE2 expressed from Expi293F cells, as well S2 subunit plant (N. benthamiana) cells. Using high-resolution Orbitrap Eclipse Tribrid mass spectrometer equipped with Ultimate 3000 RSLCnano I-GPA (Integrated GlycoProteome Analyzer) developed previous 148 28 O-glycopeptides RBD, 71 N-glycopeptides subunit, 139 were characterized. addition, report post-translational modifications (PTMs) glycan, including mannose-6-phosphate (M6P) GlcNAc-1-phosphate-6-O-mannose N-glycan RBD ACE2, O-acetylation O-glycan identified first time proteins. relative abundance distribution according glycosites glycan types analyzed quantified O (only RBD)-glycopeptides S2, using I-GPA. Asn331 Asn1098 Asn103 majorly N-glycosylated, dominant glycan-type was complex high-mannose S2. These findings will provide valuable insights into patterns that influence function immunogenicity offer new perspectives development vaccines antibody-based therapies against COVID-19.

Language: Английский

Modifying the glycosylation profile of SARS-CoV-2 spike-based subunit vaccines alters focusing of the humoral immune response in a mouse model DOI Creative Commons
Tyler M. Renner, Matthew Stuible, Martín A. Rossotti

et al.

Communications Medicine, Journal Year: 2025, Volume and Issue: 5(1)

Published: April 11, 2025

Language: Английский

Citations

0
Characterization of Site-Specific N- and O-Glycopeptides from Recombinant Spike and ACE2 Glycoproteins Using LC-MS/MS Analysis DOI Open Access

Ju Hwan Song,

S. Y. Jang,

Jinwoong Choi

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(24), P. 13649 - 13649

Published: Dec. 20, 2024

The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in hundreds of millions infections and deaths globally. Although vaccination campaigns are mitigating the emerging viral variants continue to pose challenges. spike (S) protein SARS-CoV-2 plays a critical role entry binding angiotensin-converting enzyme (ACE2) receptor, making both proteins essential targets for therapeutic vaccine development. glycosylation these influences their structure function. This underscores need detailed site-specific glycoproteomic analysis. In this study, we characterized N- or O-glycosylation profiles recombinant receptor-binding domain (RBD) ACE2 expressed from Expi293F cells, as well S2 subunit plant (N. benthamiana) cells. Using high-resolution Orbitrap Eclipse Tribrid mass spectrometer equipped with Ultimate 3000 RSLCnano I-GPA (Integrated GlycoProteome Analyzer) developed previous 148 28 O-glycopeptides RBD, 71 N-glycopeptides subunit, 139 were characterized. addition, report post-translational modifications (PTMs) glycan, including mannose-6-phosphate (M6P) GlcNAc-1-phosphate-6-O-mannose N-glycan RBD ACE2, O-acetylation O-glycan identified first time proteins. relative abundance distribution according glycosites glycan types analyzed quantified O (only RBD)-glycopeptides S2, using I-GPA. Asn331 Asn1098 Asn103 majorly N-glycosylated, dominant glycan-type was complex high-mannose S2. These findings will provide valuable insights into patterns that influence function immunogenicity offer new perspectives development vaccines antibody-based therapies against COVID-19.

Language: Английский

Citations

0