Demonstration of Antibodies Against SARS-CoV-2, Neutralizing or Binding, in Seroconversion Panels After mRNA-1273, BNT-162b2, and Ad26.COV2.S Vaccine Administration

  1. Francisco Belda
  2. Oscar Mora
  3. Monica Lopez-Martinez
  4. Nerea Torres
  5. Ana Vivanco
  6. Silvia Marfil
  7. Edwards Pradenas
  8. Marta Massanella
  9. Julià Blanco
  10. Rebecca Christie
  11. Michael Crowley

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Abstract

Vaccines are an important tool in combating the COVID-19 pandemic. Two mRNA vaccines (mRNA-1273 and BNT-162b2) and an adenovirus vector vaccine (Ad26.COV2.S) were among the first vaccines to be approved by global regulatory authorities. The aim of this observational study was to characterize the levels and time course of the generation of anti-SARS-CoV-2 spike protein antibodies after vaccination with 3 different vaccines and the neutralizing activity of these antibodies. Seroconversion panels were generated from blood samples collected before and after vaccination with 3 COVID-19 vaccines: mRNA-1273, BNT-162b2, and Ad26.COV2.S. The seroconversion panels were tested for antibody activity by chemiluminescent immunoassay or enzyme-linked immunosorbent assay (ELISA), and 1 panel was tested for neutralization activity in a pseudovirus assay. Participants positive for anti-SARS-CoV-2 antibodies before vaccination (18.6%) had a higher response to the first dose than participants who tested negative. For 2-dose vaccines, older participants showed a lower response to the first dose than younger participants. All participants showed positive responses after the second vaccine. For the adenovirus vector vaccine, 2 participants did not generate antibody responses after vaccination. Four participants were negative at 2 weeks but positive at 2 months. Pseudovirus neutralization showed good correlation with antibody activity (correlation coefficient = 0.78, P < .0001). Antibody responses in participants over 45 years old tended to be less robust. Participants that had been infected with SARS-CoV-2 and had antibodies prior to vaccination showed a more robust response to initial vaccination. Older participants (>45 years) showed less robust responses to both types of vaccine. All participants receiving full mRNA vaccination showed positive antibody responses. Some participants receiving the adenovirus vaccine did not respond. Antibody responses correlated well with neutralization activity. Seroconversion panels can be useful in the development of antibody assays and in investigating their effectiveness against new SARS-CoV-2 variants.

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  1. Version published to 10.1177/26348535231202681
  2. ScreenIT

    SciScore for 10.1101/2022.03.28.22272552: (What is this?)

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

    Table 1: Rigor

    EthicsConsent: The donors provided informed consent and their samples were collected under an approved IRB protocol ([1149706-4] Diagnostic QC and Pre-Clinical Sample Collection Project: Ballad Health System Institutional Review Board (IRB #00003204), Johnson City, TN, USA).
    IRB: The donors provided informed consent and their samples were collected under an approved IRB protocol ([1149706-4] Diagnostic QC and Pre-Clinical Sample Collection Project: Ballad Health System Institutional Review Board (IRB #00003204), Johnson City, TN, USA).
    Field Sample Permit: These studies were conducted in compliance with all applicable regulatory guidelines.
    Sex as a biological variableThere were 10 female and 5 male donors, and all were white/Caucasian.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    CLIA and ELISA Assays: The samples in the seroconversion panels were tested for the presence of anti-SARS-CoV-2 antibodies using chemiluminescent immunoassays (
    anti-SARS-CoV-2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Neutralizing Antibody Determination: Cell line: HEK293T cells (presumably of female origin) overexpressing wild-type human ACE-2 (Integral Molecular, USA) were used as target for SARS-CoV-2 spike expressing pseudovirus infection.
    HEK293T
    suggested: RRID:CVCL_HA71)
    Then 2 × 104 HEK293T/hACE2 cells treated with DEAE-Dextran (Sigma-Aldrich, St. Louis, MO, USA) were added.
    HEK293T/hACE2
    suggested: RRID:CVCL_A7UK)
    Recombinant DNA
    SentencesResources
    Pseudovirus generation: Pseudoviruses (HIV reporter) expressing the SARS-CoV-2 S protein and luciferase were constructed using the defective HIV plasmid, pNL4-3.Luc.R-.E-.
    pNL4-3.Luc.R-
    suggested: None
    The plasmid was obtained from the NIH AIDS Reagent Program. [17] Expi293F cells were transfected using ExpiFectamine293 Reagent (Thermo Fisher Scientific, Waltham, MA, USA) with pNL4-3.
    pNL4-3
    suggested: None
    Software and Algorithms
    SentencesResources
    The results were normalized and the ID50 (the reciprocal dilution inhibiting 50% of the infection) was calculated by plotting the log of plasma dilution versus response and fitting to a 4-parameter equation in Prism 8.4.3 (GraphPad Software, San Diego, CA, USA).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad
    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 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 10.1101/2022.03.28.22272552 on medRxiv