A Single Immunization with Spike-Functionalized Ferritin Vaccines Elicits Neutralizing Antibody Responses against SARS-CoV-2 in Mice

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Abstract

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  1. SciScore for 10.1101/2020.08.28.272518: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableMouse immunizations: Balb/c female mice (6-8 weeks old) were procured from The Jackson Laboratory.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    In-house made primary antibodies CR3022, COVA2-15, and CB6 (approximate concentrations 0.8 – 1.3 mg/mL) were added at a 1:10,000 dilution in PBST.
    COVA2-15
    suggested: None
    CB6
    suggested: None
    Secondary antibody was added at 1:10,000 (abcam ab6759, rabbit anti-human IgG H&L HRP) in 5% milk / PBST and incubated for 1 h at room temperature.
    anti-human IgG
    suggested: None
    For purified mAbs and ACE2-Fc, Direct-Blot HRP anti-human IgG1 Fc antibody (Biolegend 410604) was added at a 1:10,000 dilution in diluent buffer for 1 h at room temperature.
    anti-human IgG1
    suggested: (BioLegend Cat# 410604, RRID:AB_2749913)
    CCP samples were obtained from donors who had a clinical diagnosis of COVID-19 and either a positive RT-PCR or SARS-CoV-2 antibody test.
    SARS-CoV-2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    2 pseudotyped lentivirus production and viral neutralization assays: SARS-CoV-2 spike pseudotyped lentivirus was produced in HEK293T cells via calcium phosphate transfection.
    HEK293T
    suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
    Experimental Models: Organisms/Strains
    SentencesResources
    Mouse immunizations: Balb/c female mice (6-8 weeks old) were procured from The Jackson Laboratory.
    Balb/c
    suggested: None
    Software and Algorithms
    SentencesResources
    Dots were quantified using the gel analysis protocol in Fiji (ImageJ) and curves were fit using a linear regression in GraphPad Prism 8.4.1. SEC-MALS of SARS-CoV-2 antigens: SEC-MALS was performed on an Agilent 1260 Infinity II HPLC with Wyatt detectors for light scattering (miniDAWN) and refractive index (Optilab)
    Fiji
    suggested: (Fiji, RRID:SCR_002285)
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Molecular weights were determined using ASTRA 7.3.2 (Wyatt Technologies).
    ASTRA
    suggested: (ASTRA, RRID:SCR_016255)
    Single-particle image processing and 3D reconstruction: All movie stacks were first imported into Relion 3.0.6 (70) for image processing.
    Relion
    suggested: (RELION, RRID:SCR_016274)
    Motion-correction was performed with MotionCor2 1.3.2 (71) and the contrast transfer function was determined with CTFFIND4 4.1.13 (72).
    MotionCor2
    suggested: (MotionCor2, RRID:SCR_016499)
    The initial model was built in cryoSPARC (74) using the ab-initio reconstruction option with octahedral symmetry applied.
    cryoSPARC
    suggested: (cryoSPARC, RRID:SCR_016501)

    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: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT03186781CompletedInfluenza HA Ferritin Vaccine, Alone or in Prime-Boost Regim…
    NCT03814720Active, not recruitingDose, Safety, Tolerability and Immunogenicity of an Influenz…


    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.