SARS-CoV-2 Antibody Binding and Neutralization in Dried Blood Spot Eluates and Paired Plasma

  1. Hannah L. Itell
  2. Haidyn Weight
  3. Carolyn S. Fish
  4. Jennifer K. Logue
  5. Nicholas Franko
  6. Caitlin R. Wolf
  7. Denise J. McCulloch
  8. Jared Galloway
  9. Frederick A. Matsen
  10. Helen Y. Chu
  11. Julie Overbaugh

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Abstract

Plasma and sera isolated from venous blood represent conventional sample types used for the evaluation of SARS-CoV-2 antibody responses after infection or vaccination. However, collection of these samples is invasive and requires trained personnel and equipment for immediate processing.

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  1. Version published to 10.1128/spectrum.01298-21
  2. ScreenIT

    SciScore for 10.1101/2021.08.02.21261504: (What is this?)

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

    Table 1: Rigor

    EthicsField Sample Permit: Study participants: Paired DBS and plasma specimens were collected from COVID-19 convalescent and SARS-CoV-2 vaccinated individuals enrolled as part of the Hospitalized or Ambulatory Adults with Respiratory Viral Infections (HAARVI) research study in Seattle, Washington.
    Consent: All participants completed informed consent as approved by the University of Washington Institutional Review Board (protocol #STUDY00000959).
    IRB: All participants completed informed consent as approved by the University of Washington Institutional Review Board (protocol #STUDY00000959).
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Secondary antibody was prepared by diluting goat anti-human IgG-horseradish peroxidase (Sigma #A0170) to 1:2,500 in blocking buffer (2.24 µg/mL).
    anti-human IgG-horseradish
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    HEK293T cells were added to 6-well plates at 5 x 105 cells per well in DMEM supplemented with 10% fetal bovine serum, 2 mM L-glutamine, and penicillin/streptomycin/fungizone.
    HEK293T
    suggested: None
    4 HEK293T-ACE2 cells in 96-well black-walled plates and, after 16-24 hours, adding 100 µL of diluted viral supernatant per well in duplicate.
    HEK293T-ACE2
    suggested: None
    Software and Algorithms
    SentencesResources
    Duplicate OD450 measurements were averaged and IgG concentrations in plasma and DBS eluates were interpolated from the standard curve using the five-parameter logistic equation function in GraphPad Prism 8.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    The following point dilutions were assessed: 1:400 for participant plasma and negative control normal human serum (Gemini Biosciences #100-110, lot H87WOOK), 1:20 for VB DBS eluates, and 1:40 for FS DBS eluates.
    Gemini Biosciences
    suggested: (Penn Cell Center Stockroom, RRID:SCR_010003)
    Figure 1 was created in BioRender.com.
    BioRender
    suggested: (Biorender, RRID:SCR_018361)
    The remaining figures were generated using RStudio.
    RStudio
    suggested: (RStudio, RRID:SCR_000432)

    Results from OddPub: Thank you for sharing your code.

    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.

  3. Version published to 10.1101/2021.08.02.21261504v1 on medRxiv