Linear epitopes of SARS-CoV-2 spike protein elicit neutralizing antibodies in COVID-19 patients

  1. Yang Li
  2. Dan-yun Lai
  3. Hai-nan Zhang
  4. He-wei Jiang
  5. Xiaolong Tian
  6. Ming-liang Ma
  7. Huan Qi
  8. Qing-feng Meng
  9. Shu-juan Guo
  10. Yanling Wu
  11. Wei Wang
  12. Xiao Yang
  13. Da-wei Shi
  14. Jun-biao Dai
  15. Tianlei Ying
  16. Jie Zhou
  17. Sheng-ce Tao

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Abstract

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  1. Version published to 10.1038/s41423-020-00523-5
  2. ScreenIT

    SciScore for 10.1101/2020.06.07.20125096: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Patients and samples: The Institutional Ethics Review Committee of Foshan Fourth Hospital, Foshan, China approved this study and the written informed consent was obtained from each patient.
    Consent: Patients and samples: The Institutional Ethics Review Committee of Foshan Fourth Hospital, Foshan, China approved this study and the written informed consent was obtained from each patient.
    RandomizationA few conjugates were randomly selected for examination by SDS-PAGE.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Peptide microarray fabrication: The peptide-BSA conjugates as well as S1 protein, RBD protein and N protein of SARS-CoV-2, along with the negative (BSA) and positive controls (anti-Human IgG and IgM antibody), were printed in triplicate on PATH substrate slide (Grace Bio-Labs, Oregon, USA) to generate identical arrays in a 1 × 7 subarray format using Super Marathon printer (Arrayjet, UK).
    anti-Human IgG
    suggested: None
    IgM
    suggested: None
    The arrays were washed with 1×PBST and bound antibodies were detected by incubating with Cy3-conjugated goat anti-human IgG and Alexa Fluor 647-conjugated donkey anti-human IgM (Jackson ImmunoResearch, PA, USA), which were diluted for 1: 1,000 in 1×PBST.
    anti-human IgM
    suggested: None
    Antibodies or isotype IgG control (Thermo Fisher Scientific, MA, USA) in DMEM supplemented with 10% fetal calf serum were incubated with pseudoviruses at 37C for 1 h and then the mixtures were added to monolayer Huh-7 cells (104 per well in 96-well plates).
    isotype IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Huh-7 cells were subsequently lysed with 50 µl lysis reagent (Promega, WI, USA), and 30 µl of the lysates were transferred to 96-well Costar flat-bottom luminometer plates (Corning Costar, MA, USA) for the detection of relative light units using the Firefly Luciferase Assay Kit (Promega, WI, USA) and an Ultra 384 luminometer (Tecan, Switzerland).
    Huh-7
    suggested: None
    Software and Algorithms
    SentencesResources
    Pearson correlation coefficient between two proteins or indicators and the corresponding p value was calculated by SPSS software under the default parameters.
    SPSS
    suggested: (SPSS, RRID:SCR_002865)
    Cluster analysis was performed by pheatmap package in R4.
    pheatmap
    suggested: (pheatmap, RRID:SCR_016418)
    Visualization of the structural details were processed by Pymol (https://pymol.org/2/).
    Pymol
    suggested: (PyMOL, RRID:SCR_000305)

    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.

    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.

  3. Version published to 10.1101/2020.06.07.20125096v1 on medRxiv