3 research outputs found
Development of a PNGase Rc Column for Online Deglycosylation of Complex Glycoproteins during HDX-MS
Protein
glycosylation is one of the most common PTMs and many cell
surface receptors, extracellular proteins, and biopharmaceuticals
are glycosylated. However, HDX-MS analysis of such important glycoproteins
has so far been limited by difficulties in determining the HDX of
the protein segments that contain glycans. We have developed a column
containing immobilized PNGase Rc (from Rudaea cellulosilytica) that can readily be implemented into a conventional HDX-MS setup
to allow improved analysis of glycoproteins. We show that HDX-MS with
the PNGase Rc column enables efficient online removal of N-linked
glycans and the determination of the HDX of glycosylated regions in
several complex glycoproteins. Additionally, we use the PNGase Rc
column to perform a comprehensive HDX-MS mapping of the binding epitope
of a mAb to c-Met, a complex glycoprotein drug target. Importantly,
the column retains high activity in the presence of common quench-buffer
additives like TCEP and urea and performed consistent across 114 days
of extensive use. Overall, our work shows that HDX-MS with the integrated
PNGase Rc column can enable fast and efficient online deglycosylation
at harsh quench conditions to provide comprehensive analysis of complex
glycoproteins
Development of a PNGase Rc Column for Online Deglycosylation of Complex Glycoproteins during HDX-MS
Protein
glycosylation is one of the most common PTMs and many cell
surface receptors, extracellular proteins, and biopharmaceuticals
are glycosylated. However, HDX-MS analysis of such important glycoproteins
has so far been limited by difficulties in determining the HDX of
the protein segments that contain glycans. We have developed a column
containing immobilized PNGase Rc (from Rudaea cellulosilytica) that can readily be implemented into a conventional HDX-MS setup
to allow improved analysis of glycoproteins. We show that HDX-MS with
the PNGase Rc column enables efficient online removal of N-linked
glycans and the determination of the HDX of glycosylated regions in
several complex glycoproteins. Additionally, we use the PNGase Rc
column to perform a comprehensive HDX-MS mapping of the binding epitope
of a mAb to c-Met, a complex glycoprotein drug target. Importantly,
the column retains high activity in the presence of common quench-buffer
additives like TCEP and urea and performed consistent across 114 days
of extensive use. Overall, our work shows that HDX-MS with the integrated
PNGase Rc column can enable fast and efficient online deglycosylation
at harsh quench conditions to provide comprehensive analysis of complex
glycoproteins
DataSheet1_Interplay of Digital Proximity App Use and SARS-CoV-2 Vaccine Uptake in Switzerland: Analysis of Two Population-Based Cohort Studies.docx
Objectives: Our study aims to evaluate developments in vaccine uptake and digital proximity tracing app use in a localized context of the SARS-CoV-2 pandemic.Methods: We report findings from two population-based longitudinal cohorts in Switzerland from January to December 2021. Failure time analyses and Cox proportional hazards regression models were conducted to assess vaccine uptake and digital proximity tracing app (SwissCovid) uninstalling outcomes.Results: We observed a dichotomy of individuals who did not use the SwissCovid app and did not get vaccinated, and who used the SwissCovid app and got vaccinated during the study period. Increased vaccine uptake was observed with SwissCovid app use (aHR, 1.51; 95% CI: 1.40–1.62 [CI-DFU]; aHR, 1.79; 95% CI: 1.62–1.99 [CSM]) compared to SwissCovid app non-use. Decreased SwissCovid uninstallation risk was observed for participants who got vaccinated (aHR, 0.55; 95% CI: 0.38–0.81 [CI-DFU]; aHR, 0.45; 95% CI: 0.27–0.78 [CSM]) compared to participants who did not get vaccinated.Conclusion: In evolving epidemic contexts, these findings underscore the need for communication strategies as well as flexible digital proximity tracing app adjustments that accommodate different preventive measures and their anticipated interactions.</p