MassIVE MSV000088601

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Online Hydrophilic Interaction Chromatography (HILIC) Enhanced Top-Down Mass Spectrometry Characterization of SARS-CoV-2 Spike Receptor Binding Domain

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Description

Online LCMS intact glycoform separation of SARS-CoV-2 spike receptor binding domain proteins expressed in HEK-293 cells from the vendors Sino Biological or RayBiotech. C2 RPLC, CZE, and HILIC separation methods are compared as well as top-down and bottom-up analysis to define RBD glycosylation patterns. Intact proteins were buffer exchanged into 100 mM ammonium acetate before online LC separation in triplicate. ETD top-down analysis was performed after PNGase F N-glycan removal. Bottom-up analysis was performed using cysteine alkylation with iodoacetamide and trypsin digestion for RayBiotech RBDs and a combination of Glu-C and trypsin digestion for Sino Biological RBDs. Intact data (in triplicate per an RBD) were processed using PMI Intact for all glycoform separation experiments and output in csv format. Glycopeptide data were processed with PMI Byonic in the folder PMI_Byonic_results which can be found inside the directory named "other" as well as the processed intact data. [doi:10.25345/C5PS11] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: SARS-CoV-2 ; spike receptor binding domain ; glycosylation ; proteomics

Contact

Principal Investigators:
(in alphabetical order) 		Mowei Zhou, Pacific Northwest National Laboratory, United States
Submitting User: alchemistmatt

Publications

Jesse W Wilson, Aivett Bilbao, Juan Wang, Yen-Chen Liao, Dusan Velickovic, Roza Wojcik, Marta Passamonti, Rui Zhao, Andrea F G Gargano, Vincent R Gerbasi, Ljiljana Pas A-Toli?, Scott E Baker, Mowei Zhou.
Online Hydrophilic Interaction Chromatography (HILIC) Enhanced Top-Down Mass Spectrometry Characterization of the SARS-CoV-2 Spike Receptor-Binding Domain.
Anal Chem. 2022 Apr 5. doi: 10.1021/acs.analchem.2c00139.

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