Human monoclonal antibodies block the binding of SARS-CoV-2 spike protein to angiotensin converting enzyme 2 receptor

  1. Xiangyu Chen
  2. Ren Li
  3. Zhiwei Pan
  4. Chunfang Qian
  5. Yang Yang
  6. Renrong You
  7. Jing Zhao
  8. Pinghuang Liu
  9. Leiqiong Gao
  10. Zhirong Li
  11. Qizhao Huang
  12. Lifan Xu
  13. Jianfang Tang
  14. Qin Tian
  15. Wei Yao
  16. Li Hu
  17. Xiaofeng Yan
  18. Xinyuan Zhou
  19. Yuzhang Wu
  20. Kai Deng
  21. Zheng Zhang
  22. Zhaohui Qian
  23. Yaokai Chen
  24. Lilin Ye

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Abstract

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  1. ScreenIT

    SciScore for 10.1101/2020.04.06.20055475: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Human samples: The 26 COVID-19 patients enrolled in the study were provided written informed consent.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Single-cell sorting by flow cytometry: For B cell enrichment, PBMCs were firstly stained with FITC-conjugated anti-CD19 antibody (Biolegend) on ice for 30 min.
    anti-CD19
    suggested: None
    PE-conjugated anti-CD20 antibody (Biolegend),
    anti-CD20
    suggested: None
    APC-Cy7-conjugated anti-CD3 antibody (Biolegend),
    anti-CD3
    suggested: None
    anti-CD14 antibody (Biolegend),
    anti-CD14
    suggested: None
    anti-CD56 antibody (Biolegend) and APC-Cy7-conjugated LIVE/DEAD dye (Life Technologies).
    anti-CD56
    suggested: None
    After washing with PBST, the bound antibodies were incubated with anti-human IgG HRP detection antibody (Bioss Biotech) for 30 min, followed by washed with PBST, then PBS and addition of TMB (Beyotime).
    anti-human IgG
    suggested: None
    Then, the incubated mixtures were added to ELISA plates and allowed to develop for 30 min, followed by PBST washing and anti-mouse Fc HRP antibody (Thermo Fisher Scientific).
    anti-mouse Fc HRP
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The digested heavy and light chain genes were further cloned into human IgG1 heavy chain and light chain expression vectors, respectively. Transfection: Human embryonic kidney (HEK) 293T cells of 80-90% confluent in the 15 cm tissue culture plate were transfected with master mixture containing 9 μg heavy chain plasmid, 9 μg light chain plasmid and 60 μl
    HEK
    suggested: None
    Then, the mixtures were incubated with 10,000 hACE2-plasmid transiently transfected 293T cells for 40 min on ice, followed by stained with Alexa Fluor 647-conjugated goat anti-mouse IgG (Biolegend) and APC-Cy7-conjugated LIVE/DEAD dye (Life Technologies).
    293T
    suggested: None
    Concisely, HEK-293T cells were transfected with psPAX2, pLenti-GFP and 2019-nCov S plasmids by using TransIT-293 Transfection reagent (Mirus).
    HEK-293T
    suggested: None
    At 40 hours-post incubation, the luciferase activity of infected hACE2/293T cells were detected by Dual-Luciferase Reporter Assay System (
    hACE2/293T
    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.
    • 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 10.1038/s41423-020-0426-7
  3. Version published to 10.1101/2020.04.06.20055475v1 on medRxiv