Recombinant SARS-CoV-2 spike proteins for sero-surveillance and epitope mapping

  1. Sophie M. Jegouic
  2. Silvia Loureiro
  3. Michelle Thom
  4. Deepa Paliwal
  5. Ian M. Jones

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Abstract

The newly emergent SARS-CoV-2 coronavirus is closely related to SARS-CoV which emerged in 2002. Studies on coronaviruses in general, and SARS in particular, have identified the virus spike protein (S) as being central to virus tropism, to the generation of a protective antibody response and to the unambiguous detection of past infections. As a result of this centrality SARS-CoV-2 S protein has a role in many aspects of research from vaccines to diagnostic tests. We describe a number of recombinant forms of SARS-CoV-2 S expressed in commonly available expression systems and their preliminary use in diagnostics and epitope mapping. These sources may find use in the current and future analysis of the virus and the Covid-19 disease it causes.

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  1. ScreenIT

    SciScore for 10.1101/2020.05.21.109298: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The plates were washed five times with TBS containing 0.05% v/v Tween-20 and polyclonal anti-human antibody conjugated to HRP (Sigma) added for one hour at room temperature.
    anti-human
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cell culture: Spodoptera frugiperda (Sf9) and T.niao38 cells were maintained in EX-CELL 420 medium (Sigma) supplemented with 2% fetal bovine serum, 1% penicillin/streptomycin solution, at 27°C with shaking.
    T.niao38
    suggested: None
    Software and Algorithms
    SentencesResources
    Alternate hosts used were SoluBL21 (AMS Biotechnology) and LOBSTR [17].
    LOBSTR
    suggested: (lobSTR, RRID:SCR_008030)
    Protein Purification: Infected insect cells were disrupted with CytoBuster™ protein extraction reagent (Merck) and clarified by centrifugation at 4,300 × g for 20m before column loading.
    CytoBuster™
    suggested: None

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    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.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    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.
    • No funding statement was detected.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.05.21.109298v1 on bioRxiv