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Identification, mapping and relative quantitation of SARS-CoV-2 Spike glycopeptides by Mass-Retention Time Fingerprinting.
Chalk, Rod; Greenland, William E P; Moreira, Tiago; Coker, Jesse; Mukhopadhyay, Shubhashish M M; Williams, Eleanor; Manning, Charlotte; Bohstedt, Tina; McCrorie, Rama; Fernandez-Cid, Alejandra; Burgess-Brown, Nicola A.
  • Chalk R; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK. rod.chalk@cmd.ox.ac.uk.
  • Greenland WEP; Agilent Technologies, Lakeside, Cheadle Royal Business Park, Cheadle, Cheshire, UK.
  • Moreira T; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • Coker J; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • Mukhopadhyay SMM; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • Williams E; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • Manning C; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • Bohstedt T; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • McCrorie R; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • Fernandez-Cid A; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
  • Burgess-Brown NA; Centre for Medicines Discovery, ORCRB, University of Oxford, Oxford, UK.
Commun Biol ; 4(1): 934, 2021 08 03.
Article in English | MEDLINE | ID: covidwho-1341013
Preprint
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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.
Subject(s)

Full text: Available Collection: International databases Database: MEDLINE Main subject: Mass Spectrometry / Glycopeptides / Spike Glycoprotein, Coronavirus Language: English Journal: Commun Biol Year: 2021 Document Type: Article Affiliation country: S42003-021-02455-w

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Mass Spectrometry / Glycopeptides / Spike Glycoprotein, Coronavirus Language: English Journal: Commun Biol Year: 2021 Document Type: Article Affiliation country: S42003-021-02455-w