Minimal system for assembly of SARS-CoV-2 virus like particles

  1. Heather Swann
  2. Abhimanyu Sharma
  3. Benjamin Preece
  4. Abby Peterson
  5. Crystal Eldredge
  6. David M. Belnap
  7. Michael Vershinin
  8. Saveez Saffarian

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Abstract

SARS-CoV-2 virus is the causative agent of COVID-19. Here we demonstrate that non-infectious SARS-CoV-2 virus like particles (VLPs) can be assembled by co-expressing the viral proteins S, M and E in mammalian cells. The assembled SARS-CoV-2 VLPs possess S protein spikes on particle exterior, making them ideal for vaccine development. The particles range in shape from spherical to elongated with a characteristic size of 129 ± 32 nm. We further show that SARS-CoV-2 VLPs dried in ambient conditions can retain their structural integrity upon repeated scans with Atomic Force Microscopy up to a peak force of 1 nN.

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  1. Version published to 10.1038/s41598-020-78656-w
  2. Version published to 10.1101/2020.06.01.128058v3 on bioRxiv
  3. Version published to 10.1101/2020.06.01.128058v2 on bioRxiv
  4. ScreenIT

    SciScore for 10.1101/2020.06.01.128058: (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
    VLPs were incubated with 1:1000 dilution of anti-S antibody (clone 1A9, GTX632604, GeneTex, Irvine, CA) and 1:5 dilution of goat anti-mouse IgG nanogold conjugates (BBI solutions, Crumlin, UK, distributed by TedPella, Redding, CA, cat # 15753).
    anti-mouse IgG
    suggested: (Creative Diagnostics Cat# DCABH-15753, RRID:AB_2487965)
    Membranes were stained with 1:1000 dilution of Anti-SARS-CoV SΔ10 antibody ([1A9], GenTex, Irvine, CA) along with 1:1000 dilution of Anti-Membrane Protein (2019-nCoV
    Anti-SARS-CoV SΔ10
    suggested: None
    Anti-Membrane Protein (2019-nCoV
    suggested: None
    Glass coverslips were functionalized with anti-S antibody as follows.
    anti-S
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    After triple washing the cells with PBS, 293T cells were suspended in 100ul of RIPA buffer (sc-24948, Santa Cruz, Dallas, Texas).
    293T
    suggested: None
    Software and Algorithms
    SentencesResources
    The surfaces were then incubated with neutravidin (Thermo Scientific Pierce Protein Biology, Waltham, MA, USA) followed by a brief dd-H20 wash to get rid of excess neutravidin and incubation with biotinylated anti-S antibody (11-2001-B, Abeomics, San Diego, CA, USA)
    Thermo Scientific Pierce Protein Biology
    suggested: None

    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.

  5. Version published to 10.1101/2020.06.01.128058v1 on bioRxiv