Identification, Mapping and Relative Quantitation of SARS-CoV-2 Spike Glycopeptides by Mass-Retention Time Fingerprinting

  1. R. Chalk
  2. W. Greenland
  3. T. Moreira
  4. J. Coker
  5. S.M.M Mukhopadhyay
  6. E. Williams
  7. C. Manning
  8. T. Bohstedt
  9. R. McCrorie
  10. A. Fernandez-Cid
  11. N.A. Burgess-Brown

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Abstract

We describe a novel analytical method for rapid and robust identification, mapping and relative quantitation of glycopeptides from SARS-CoV-2 Spike protein. The method may be executed using any LC-TOF mass spectrometer, requires no specialised knowledge of glycan analysis and makes use of the differential resolving power of reversed 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. While this has previously been viewed as 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. This database allows any ESI-TOF equipped lab to reliably identify and quantify spike glycans from a single overnight elastase protein digest in less than 90 minutes.

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    SciScore for 10.1101/2020.07.24.217562: (What is this?)

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    Table 2: Resources

    Software and Algorithms
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    This is possible either using the Agilent software described, software provided by other vendors, or by manual inspection.
    Agilent
    suggested: (Agilent Bravo NGS, RRID:SCR_019473)

    Results from OddPub: Thank you for sharing your data.

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

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    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
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    • No protocol registration statement was detected.

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  2. Version published to 10.1101/2020.07.24.217562 on bioRxiv