A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors

  1. Xinjian Yin
  2. Litong Chen
  3. Siwen Yuan
  4. Lan Liu
  5. Zhizeng Gao

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Abstract

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  1. Version published to 10.1016/j.bioorg.2021.105362
  2. ScreenIT

    SciScore for 10.1101/2021.06.17.448891: (What is this?)

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

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    Recombinant DNA
    SentencesResources
    Ltd (Guangzhou, China) and inserted into plasmid pET-44b(+) and pET-28a(+), respectively.
    pET-44b(+
    suggested: None
    pET-28a(+)
    suggested: RRID:Addgene_108949)
    2.2 Protein expression and purification: The plasmids pET-44b-NusA-5HB, pET-28a-5HB, and pCold-NusA containing the gene encoding NusA-5HB, 5HB and NusA (with N-terminal His-tag) were transformed into competent E. coli BL21 (DE3) cells (Invitrogen) respectively.
    pET-28a-5HB
    suggested: None
    pCold-NusA
    suggested: None
    The recombinant E. coli was cultivated in LB medium with antibiotic (50 mg/L Kan for pET-28a-5HB; 100 mg/L Amp for pET-44b-NusA-5HB and pCold-NusA) at 37 °C and rotary shaking at 200 rpm.
    pET-44b-NusA-5HB
    suggested: None
    Software and Algorithms
    SentencesResources
    The obtained f values were plotted using GraphPad Prism 8 to calculate the IC50.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    PyMOL (Version 1.5, Schrö dinger, LLC) was used to prepare figures of the complexes.
    PyMOL
    suggested: (PyMOL, RRID:SCR_000305)

    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.

    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 10.1101/2021.06.17.448891v1 on bioRxiv