1. SciScore for 10.1101/2021.06.08.447530: (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.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Anti-M13-HRP antibody (Sino Biological) was used for detection at 1:4000 dilution in blocking buffer.
    suggested: None
    Plates were blocked for 1 hr at RT with 5% bovine serum albumin (BSA) in PBS before a 1 hr incubation with serially diluted VNAR-hFc antibodies or ACE2.
    suggested: None
    The plates were washed, and binding was detected with an anti-human IgG (Fc specific) (1:5000 dilution, Sigma-Aldrich, A0170) or anti-FLAG (1:1000 dilution, Sigma-Aldrich, A8592) HRP-conjugated antibodies for VNAR-hFc antibodies or ACE2, respectively.
    anti-human IgG
    suggested: (Sigma-Aldrich Cat# A0170, RRID:AB_257868)
    suggested: (Sigma-Aldrich Cat# A8592, RRID:AB_439702)
    ACE2 binding was measured with an anti-FLAG HRP conjugated antibody (Sigma-Aldrich, A8592) diluted 1:1000.
    anti-FLAG HRP
    suggested: None
    suggested: (Sigma-Aldrich Cat# A8592, RRID:AB_439702)
    After blocking in 5% BSA in PBS for 1 hr, 50 nM biotinylated S1-RBD alone or premixed with 500 nM secondary (competitor) VNAR-hFc antibodies were added to the plate pairwise.
    suggested: None
    Experimental Models: Cell Lines
    Expi293 cells were transiently transfected as VNAR human Fc fusions (VNAR-hFc) and produced following the manufacturer’s manual.
    suggested: RRID:CVCL_D615)
    Vero CCL81 cells were seeded at a cell density of 100,000 cells per well in 48-well plates and incubated at 37°C in serum free OptiPro SFM (Thermo Fisher Scientific) for 24 hr before infection.
    Vero CCL81
    suggested: None
    Recombinant DNA
    The ACE2 (UniProt entry: Q9BYF1) ectodomain (18-740) was synthesized with FLAG and 6xHis tags at the C-terminal end separated by a single G4S linker before cloning into the pFUSE vector.
    suggested: None
    Flow cytometry: Expi293 cells were transiently transfected with ACE2 cloned into the pCMV3-C-GFPSpark expression vector (Sino Biological).
    suggested: None
    Software and Algorithms
    After gating on cells expressing ACE2-GFP fusion protein in FL1 channel, median fluorescence intensity (MFI) of cells positive for S1 binding was used to determine IC50 by 4-parametric non-linear regression analysis using GraphPad Prism 8.0.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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.

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