A stable platform for the production of virus-like particles pseudotyped with the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) spike protein

  1. Sylvie Roy
  2. Karim Ghani
  3. Pedro O. de Campos-Lima
  4. Manuel Caruso

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Abstract

No abstract available

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  1. Version published to 10.1016/j.virusres.2021.198305
  2. ScreenIT

    SciScore for 10.1101/2020.09.16.298992: (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 variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    A human chimeric anti-S1 antibody (Genscript; 1:200 dilution) followed by an Alexa647-conjugated goat anti-human IgG (Jackson Laboratories; 1:400) were successively incubated with cells for labelling.
    anti-S1
    suggested: None
    anti-human IgG
    suggested: None
    Detached cells were labelled with a mouse anti-ACE2 antibody (R&D Systems, Minneapolis, MN1/200) followed by an Alexa488 goat anti-mouse (1:1,000; Invitrogen, Carlsbad, CA).
    anti-ACE2
    suggested: None
    anti-mouse
    suggested: None
    Immunoblotting was performed with a rabbit polyclonal antibody anti-S2 (1:400 dilution, SinoBiological, Beijing, China) and a rat monoclonal antibody anti-MLV p30 produced from the hybridoma R187 (1:2,000 dilution; American Type Culture Collection, Manassas, VA).
    anti-S2
    suggested: None
    anti-MLV
    suggested: (Creative Diagnostics Cat# DCABH-2412, RRID:AB_2476319)
    Blots were then incubated with secondary antibodies IRDylight680 goat anti-rat IgG (1:10,000; Invitrogen) and IRDye 800CW anti-rabbit IgG (1:10,000; Li-Cor Biosciences, Lincoln, NE), and analyzed with the Odyssey Infrared Imaging System (Li-Cor Biosciences).
    anti-rat IgG
    suggested: None
    anti-rabbit IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cell Lines: 293GP, 293 cells (ATCC, CRL-11268), and their derivatives expressing the ACE2 receptor (293-ACE2), S (293GP-S and 293-S), DeltaS (293GP-ΔS and 293-ΔS) and the Galv envelope (293GP-Galv) were cultured with Dulbecco’s modified Eagle’s medium (DMEM; Wisent, Canada) supplemented with 10% fetal calf serum (Life Technologies, Grand island, NY) and antibiotics (Wisent).
    293
    suggested: RRID:CVCL_DR94)
    Recombinant viruses from stable 293GP-S/GFP, 293GP-ΔS/GFP and 293GP-Galv/GFP cells were also produced similarly in 60-mm dishes.
    293GP-Galv/GFP
    suggested: None
    Syncytia Formation Assay: 293-ACE2 cells were mixed with 293, 293GP-S and 293GP-ΔS at a 9/1 ratio and plated at 4 × 105 cells/well in a 24-well plate.
    293-ACE2
    suggested: RRID:CVCL_DR94)
    The presence of S released in the supernatant of transiently transfected 293GP cells was analyzed by Western blot.
    293GP
    suggested: RCB Cat# RCB2354, RRID:CVCL_E072)
    Supernatants from confluent 293GP-S, 293GP-ΔS, 293-S and 293-ΔS cells were also harvested and concentrated from 60-mm dishes.
    293-ΔS
    suggested: None
    Software and Algorithms
    SentencesResources
    Plasmids: The expression plasmid pMD2ACE2iPuror containing the human angiotensin-converting enzyme (ACE2) cDNA used to generate ACE2 positive cells was constructed as follows: the ACE2 PmeI cDNA fragment obtained from the plasmid hACE2 (Addgene; #1786) was cloned in pMD2iPuror opened in EcoRV.
    Addgene
    suggested: (Addgene, RRID:SCR_002037)
    The fluorescence intensity of infected cells displayed in figure 5A were scanned using the Fiji software to evaluate the difference in viral titers (66).
    Fiji
    suggested: (Fiji, RRID:SCR_002285)
    Serial dilutions of known amounts of C-terminally Fc-tagged S2 (BioVendor, Brno, Czech Republic) were used for quantification.
    BioVendor
    suggested: (BioVendor Laboratory Medicine, RRID:SCR_005143)

    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.

  3. Version published to 10.1101/2020.09.16.298992v1 on bioRxiv