Inhibitor screening using microarray identifies the high capacity of neutralizing antibodies to Spike variants in SARS-CoV-2 infection and vaccination

  1. Xiaomei Zhang
  2. Mei Zheng
  3. Hongye Wang
  4. Haijian Zhou
  5. Te Liang
  6. Jiahui Zhang
  7. Jing Ren
  8. Huoying Peng
  9. Siping Li
  10. Haodong Bian
  11. Chundi Wei
  12. Shangqi Yin
  13. Chaonan He
  14. Ying Han
  15. Minghui Li
  16. Xuexin Hou
  17. Jie Zhang
  18. Liangzhi Xie
  19. Jing Lv
  20. Biao Kan
  21. Yajie Wang
  22. Xiaobo Yu

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Abstract

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  1. Version published to 10.7150/thno.67038
  2. ScreenIT

    SciScore for 10.1101/2021.05.15.21257254: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: Our research was approved by the Ethics Committee of Beijing Ditan Hospital (No. 2021-010-01), and exemption of informed consent was obtained prior to sera collection.
    Consent: Our research was approved by the Ethics Committee of Beijing Ditan Hospital (No. 2021-010-01), and exemption of informed consent was obtained prior to sera collection.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The highest dilution of serum that showed complete inhibition activity of SARS-CoV-2 was recorded as the neutralizing antibody titer (ID50).
    ID50
    suggested: (bNAber Cat# bNAberID_50, RRID:AB_2491067)
    After washing three times with PBST, the array was then incubated for 20 min with a Cy3 labelled donkey anti-human IgG(H+L) antibody (Jackson ImmunoResearch, USA) (2 μg/mL).
    anti-human IgG(H+L
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The proteins were expressed in HEK293 cells, followed by further purification on a Ni-NTA spin column (Thermo Fisher)
    HEK293
    suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
    The mixture was placed into a 96-well plate and incubated at 37°C for 2 h in CO2 5%, and then 2□×□104 Vero cells were added and incubated again at 37°C.
    Vero
    suggested: None
    Software and Algorithms
    SentencesResources
    Statistical analyses were performed using the GraphPad Prism software 8.3 and Microsoft Excel with the unpaired t test and Mann-Whitney test.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Microsoft Excel
    suggested: (Microsoft Excel, RRID:SCR_016137)

    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: We detected the following sentences addressing limitations in the study:
    There were several limitations in this study. First, the mSAIS assay is in vitro, which might not accurately reflect results in vivo. Second, some conformational and glycosylation epitopes might not be detected using the peptide microarray that employed chemically-synthesized peptides (Wang et al., 2020). Finally, the number of serum samples from convalescent COVID-19 patients and vaccinees were limited. The results obtained in this study should be validated in a large different cohort in the future.

    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.

    Results from scite Reference Check: We found no unreliable references.

    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 10.1101/2021.05.15.21257254v1 on medRxiv