1. Identification, mapping and relative quantitation of SARS-CoV-2 Spike glycopeptides by Mass-Retention Time Fingerprinting
- Author
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William E. P. Greenland, Nicola A. Burgess-Brown, Tina Bohstedt, Eleanor Williams, Rod Chalk, Charlotte Manning, Rama McCrorie, Tiago Moreira, Alejandra Fernández-Cid, Jesse Coker, and S.M.M. Mukhopadhyay
- Subjects
Glycan ,Time Factors ,QH301-705.5 ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Glycobiology ,Medicine (miscellaneous) ,Peptide ,Computational biology ,Mass spectrometry ,General Biochemistry, Genetics and Molecular Biology ,Article ,Biology (General) ,Databases, Protein ,chemistry.chemical_classification ,biology ,Chemistry ,Glycopeptides ,Glycopeptide ,Identification (information) ,Spike Glycoprotein, Coronavirus ,biology.protein ,Spike (software development) ,General Agricultural and Biological Sciences ,Retention time - Abstract
We describe an analytical method for the identification, mapping and relative quantitation of glycopeptides from SARS-CoV-2 Spike protein. The method may be executed using a LC-TOF mass spectrometer, requires no specialized knowledge of glycan analysis and exploits the differential resolving power of reverse phase HPLC. While this separation technique resolves peptides with high efficiency, glycans are resolved poorly, if at all. Consequently, glycopeptides consisting of the same peptide bearing different glycan structures will all possess very similar retention times and co-elute. Rather than a disadvantage, we show that shared retention time can be used to map multiple glycan species to the same peptide and location. In combination with MSMS and pseudo MS3, we have constructed a detailed mass-retention time database for Spike glycopeptides. This database allows any accurate mass LC-MS laboratory to reliably identify and quantify Spike glycopeptides from a single overnight elastase digest in less than 90 minutes., Mapping multiple glycan species to the same peptide and location from shared retention time of glycopeptides with MS and HPLC, Chalk, Greenland et al. construct a detailed mass-retention time database for Spike protein glycopeptides. This allows any mass LC-MS laboratory to reliably identify and quantify Spike protein glycopeptides from a single overnight elastase protein digest in less than 90 minutes.
- Published
- 2021