Monoclonal antibodies for the S2 subunit of spike of SARS-CoV cross-react with the newly-emerged SARS-CoV-2

  1. Zhiqiang Zheng
  2. Vanessa M. Monteil
  3. Sebastian Maurer-Stroh
  4. Chow Wenn Yew
  5. Carol Leong
  6. Nur Khairiah Mohd-Ismail
  7. Suganya Cheyyatraivendran Arularasu
  8. Vincent Tak Kwong Chow
  9. Raymond Lin Tzer Pin
  10. Ali Mirazimi
  11. Wanjin Hong
  12. Yee-Joo Tan

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Abstract

The emergence of a novel coronavirus, SARS-CoV-2, at the end of 2019 has resulted in widespread human infections across the globe. While genetically distinct from SARS-CoV, the etiological agent that caused an outbreak of severe acute respiratory syndrome (SARS) in 2003, both coronaviruses exhibit receptor binding domain (RBD) conservation and utilize the same host cell receptor, angiotensin-converting enzyme 2 (ACE2), for virus entry. Therefore, it will be important to test the cross-reactivity of antibodies that have been previously generated against the surface spike (S) glycoprotein of SARS-CoV in order to aid research on the newly emerged SARS-CoV-2. Here, we show that an immunogenic domain in the S2 subunit of SARS-CoV S is highly conserved in multiple strains of SARS-CoV-2. Consistently, four murine monoclonal antibodies (mAbs) raised against this immunogenic SARS-CoV fragment were able to recognise the S protein of SARS-CoV-2 expressed in a mammalian cell line. Importantly, one of them (mAb 1A9) was demonstrated to detect S in SARS-CoV-2-infected cells. To our knowledge, this is the first study showing that mAbs targeting the S2 domain of SARS-CoV can cross-react with SARS-CoV-2 and this observation is consistent with the high sequence conservation in the S2 subunit. These cross-reactive mAbs may serve as tools useful for SARS-CoV-2 research as well as for the development of diagnostic assays for its associated coronavirus disease COVID-19.

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  1. ScreenIT

    SciScore for 10.1101/2020.03.06.980037: (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
    Cells were immunolabelled for 1 hour at RT with the indicated murine mAb and 45 min with Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Life Technologies).
    anti-mouse IgG
    suggested: None
    They were then washed 3 times with PBS and incubated with goat anti-mouse conjugated to Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher) containing DAPI in immunofluorescence buffer for an additional incubation of 30 minutes.
    anti-mouse
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cells: Vero E6 and COS-7 cells were purchased from American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Scientific) supplemented with 10% foetal bovine serum (FBS; HyClone) and penicillin-streptomycin solution (Thermo Fisher Scientific).
    Vero E6
    suggested: None
    COS-7
    suggested: None
    Vero-E6 cells were infected with SARS-CoV-2 (SARS-CoV-2-Iso/01/human/2020/SWE, GenBank accession no. MT093571) at a multiplicity of infection (MOI) of 1 in DMEM 2% FBS (Thermo Fisher).
    Vero-E6
    suggested: None
    Software and Algorithms
    SentencesResources
    A multiple sequence alignment was created with MAFFT using the slow but accurate L-INS-I parameter settings [21] and the alignment curated, cut to the target region 1029-1192 (SARS-CoV numbering) and visualized with Jalview [22].
    MAFFT
    suggested: (MAFFT, RRID:SCR_011811)
    Jalview
    suggested: (Jalview, RRID:SCR_006459)
    The nucleotide sequences were searched with BLASTX against the reference spike glycoprotein.
    BLASTX
    suggested: (BLASTX, RRID:SCR_001653)

    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.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • 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 10.1101/2020.03.06.980037 on bioRxiv