Neutralizing antibody against SARS-CoV-2 spike in COVID-19 patients, health care workers, and convalescent plasma donors

  1. Cong Zeng
  2. John P. Evans
  3. Rebecca Pearson
  4. Panke Qu
  5. Yi-Min Zheng
  6. Richard T. Robinson
  7. Luanne Hall-Stoodley
  8. Jacob Yount
  9. Sonal Pannu
  10. Rama K. Mallampalli
  11. Linda Saif
  12. Eugene Oltz
  13. Gerard Lozanski
  14. Shan-Lu Liu

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Abstract

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  1. ScreenIT

    SciScore for 10.1101/2020.08.02.20166819: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    BlindingPlasma and serum were collected from hospitalized COVID-19 inpatients or ICU patients, OSU health care workers, and blinded convalescent plasma donors and analyzed in a blinded manner.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Antibodies used for Western blotting included anti-C9 (anti-rhodopsin) (Santa Cruz, #57432), anti-p24 (abcam, #ab63917), anti-β-actin (Sigma, #A1978) and secondary antibodies anti-Mouse IgG (Sigma, #A5278)
    anti-C9
    suggested: None
    anti-C9 ( anti-rhodopsin )
    suggested: None
    anti-p24
    suggested: None
    anti-β-actin
    suggested: None
    anti-Mouse IgG
    suggested: (Sigma-Aldrich Cat# A5278, RRID:AB_258232)
    Secondary antibodies used for flow cytometry included FITC-conjugated anti-human IgG-Fc (Sigma,405 #F9512).
    anti-human IgG-Fc
    suggested: None
    After three washes with PBS plus 2% FBS, cells were incubated with FITC-conjugated anti-human IgG (1:200, Sigma, #F0257) secondary antibodies for 1 hr.
    anti-human IgG
    suggested: None
    Following wash steps, wells were treated with 100 μl of HRP labeled anti-human-IgG tracer antibody (EDI, #31220), incubated at room temperature for 30 min, and again washed.
    anti-human-IgG tracer
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    HEK293T (ATCC CRL-11268, RRID: CVCL_1926), HeLa (ATCC, CCL-2, RRID: CVCL_0030), HTX (a subclone of HT1080), A549 (ATCC, CCL-185, RRID: CVCL_0023) and Huh7.5 (RRID: CVCL_7927) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 1% penicillin/streptomycin and 10% (vol/vol) FBS.
    HEK293T
    detected: (ATCC Cat# CRL-11268, RRID:CVCL_1926)
    HeLa
    detected: (BCRC Cat# 60005, RRID:CVCL_0030)
    HT1080
    suggested: None
    A549
    suggested: None
    HTX a
    detected: (BCRC Cat# 60074, RRID:CVCL_0023)
    Huh7.5
    detected: ( RRID:CVCL_7927)
    Calu-3 (ATCC, gift of Stelle Cormet-Boyaka) were grown in Eagle’s Minimum Essential Medium (EMEM), supplemented with 1% penicillin/streptomycin and 10% (vol/vol) FBS.
    Calu-3
    suggested: KCLB Cat# 30055, RRID:CVCL_0609)
    HEK293T/ACE2 cell line is a gift from Dr. Fang Li at the University of Minnesota. HeLa, A549, HTX and Huh7.5 cells stably expressing ACE2 were generated by transduction of pLenti-GFP vectors expressing ACE2 (OriGene, #RC208442L4), followed by puromycin selection (1 μg/mL) for 6 days.
    HEK293T/ACE2
    suggested: None
    HeLa
    suggested: None
    Huh7.5
    suggested: RRID:CVCL_7927)
    Production of intron-Gluc- or secNluc-based lentiviral pseudotypes bearing the S protein of SARS Coronaviruses and viral infection: For intron-Gluc- or secNluc-based pseudotyped lentiviral production, we transfected HEK293T cells with HIV-1-NL4.3 inGluc or secNluc vector plus a plasmid expressing the S protein or VSV-G in a 2:1 ratio using polyethylenimine (PEI).
    HEK293T
    suggested: None
    Serum/virus mixtures were then used to infect confluent Vero-E6 cells for 1 h at 37°C.
    Vero-E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Statistical analyses were performed using GraphPad Prism 5.0 as follows: One-way Analysis of Variance (ANOVA) with Bonferroni’s post-tests was used to compute statistical significance between multiple groups for multiple comparison.
    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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    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.

  2. Version published to 10.1172/jci.insight.143213
  3. Version published to 10.1101/2020.08.02.20166819v1 on medRxiv