Preservation of neutralizing antibody function in COVID‐19 convalescent plasma treated using a riboflavin and ultraviolet light‐based pathogen reduction technology

  1. Susan Yonemura
  2. Lindsay Hartson
  3. Taru S. Dutt
  4. Marcela Henao‐Tamayo
  5. Raymond Goodrich
  6. Susanne Marschner

Reviewed by

Read the full article

Listed in

Log in to save this article

Abstract

Convalescent plasma (CP) has been embraced as a safe therapeutic option for coronavirus disease 2019 (COVID‐19), while other treatments are developed. Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is not transmissible by transfusion, but bloodborne pathogens remain a risk in regions with high endemic prevalence of disease. Pathogen reduction can mitigate this risk; thus, the objective of this study was to evaluate the effect of riboflavin and ultraviolet light (R + UV) pathogen reduction technology on the functional properties of COVID‐19 CP (CCP).

Materials and methods

COVID‐19 convalescent plasma units ( n  = 6) from recovered COVID‐19 research donors were treated with R + UV. Pre‐ and post‐treatment samples were tested for coagulation factor and immunoglobulin retention. Antibody binding to spike protein receptor‐binding domain (RBD), S1 and S2 epitopes of SARS‐CoV‐2 was assessed by ELISA. Neutralizing antibody (nAb) function was assessed by pseudovirus reporter viral particle neutralization (RVPN) assay and plaque reduction neutralization test (PRNT).

Results

Mean retention of coagulation factors was ≥70%, while retention of immunoglobulins was 100%. Starting nAb titres were low, but PRNT 50 titres did not differ between pre‐ and post‐treatment samples. No statistically significant differences were detected in levels of IgG ( P  ≥ 0·3665) and IgM ( P  ≥ 0·1208) antibodies to RBD, S1 and S2 proteins before and after treatment.

Conclusion

R + UV PRT effects on coagulation factors were similar to previous reports, but no significant effects were observed on immunoglobulin concentration and antibody function. SARS‐CoV‐2 nAb function in CCP is conserved following R + UV PRT treatment.

Article activity feed

  1. Version published to 10.1111/vox.13108
  2. ScreenIT

    SciScore for 10.1101/2021.02.18.21251437: (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
    CCP units were analyzed for selected coagulation factors, immunoglobulins, and SARS-CoV-2 antibody binding and neutralizing activity.
    SARS-CoV-2
    suggested: None
    Functional Assays: An enzyme-linked immunosorbent assay (ELISA) was performed at Colorado State University to test CCP samples and a negative control (normal plasma sample) for antibody binding to the SARS-CoV-2 spike protein receptor-binding domain (RBD) and epitopes associated with the spike protein subunits S1 and S2 (catalog numbers 40592-V08H, 40591-V08H, and 40590-V08B, Sino Biological US Inc., Wayne, PA, USA).
    SARS-CoV-2 spike protein receptor-binding domain ( RBD )
    suggested: None
    S2
    suggested: None
    40591-V08H
    suggested: None
    Plates were then washed and incubated for 1 hour with horseradish peroxidase (HRP) conjugated anti-human IgG or anti-human IgM secondary antibodies (Jackson ImmunoResearch Inc.) prepared in blocking buffer (1:10,000 dilution).
    anti-human IgG
    suggested: None
    anti-human IgM
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Greiner Bio One) containing recently confluent Vero cells (ATCC, Manassas, VA, USA) were inoculated with the virus-plasma mixtures.
    Vero
    suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
    Software and Algorithms
    SentencesResources
    Statistical analysis was performed using Prism 8 for Windows (GraphPad Software, Inc.
    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: We detected the following sentences addressing limitations in the study:
    Limitations of this study include the small sample size and the generally low anti-SARS-CoV-2 titers in the CCP units. In an Emergency Use Authorization for the use of CCP to treat hospitalized COVID-19 patients, the United States Food and Drug Administration defined high-titer CCP to be units with a signal-to-cutoff value of 12 or greater when tested by the Ortho VITROS SARS-CoV-2 IgG test, which corresponds to an ID50 titer cutoff of 250 using a SARS-CoV-2 neutralization assay similar to the PRNT [38]. The six CCP units evaluated in this study were collected specifically for research at a time when blood centers were urgently calling for therapeutic CCP donations. It is possible that the donors providing research CCP units were unable to donate therapeutic units due to low antibody titers or other donor deferral factors. Despite the low titers, the various antibody assays performed in this study consistently demonstrated stability between pre- and post-treatment samples, whether testing for retention, epitope binding, or neutralizing activity.

    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.
    • No funding statement was detected.
    • 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 10.1101/2021.02.18.21251437v1 on medRxiv