Plasma From Recovered COVID-19 Patients Inhibits Spike Protein Binding to ACE2 in a Microsphere-Based Inhibition Assay
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
We present a microsphere-based flow cytometry assay that quantifies the ability of plasma to inhibit the binding of spike protein to angiotensin-converting enzyme 2. Plasma from 22 patients who had recovered from mild coronavirus disease 2019 (COVID-19) and expressed anti–spike protein trimer immunoglobulin G inhibited angiotensin-converting enzyme 2–spike protein binding to a greater degree than controls. The degree of inhibition was correlated with anti–spike protein immunoglobulin G levels, neutralizing titers in a pseudotyped lentiviral assay, and the presence of fever during illness. This inhibition assay may be broadly useful to quantify the functional antibody response of patients recovered from COVID-19 or vaccine recipients in a cell-free assay system.
Article activity feed
-
-
SciScore for 10.1101/2020.06.09.20127050: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Samples from negative controls ranged in age from 19 to 66 years of age (median 37) and 18/30 were female (60%). Table 2: Resources
Antibodies Sentences Resources Successful coupling was confirmed by staining ∼0.5 μL coupled beads with PE-conjugated anti-HIS tag antibodies (Santa Cruz, CAT #8036, USA) and compared to positive and negative controls by flow cytometry. anti-HIS tagsuggested: NoneSoftware and Algorithms Sentences Resources Statistical Analyses: Technical replicates were averaged and data were imported to Prism 8.0 (GraphPad). GraphPadsuggested: …SciScore for 10.1101/2020.06.09.20127050: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Samples from negative controls ranged in age from 19 to 66 years of age (median 37) and 18/30 were female (60%). Table 2: Resources
Antibodies Sentences Resources Successful coupling was confirmed by staining ∼0.5 μL coupled beads with PE-conjugated anti-HIS tag antibodies (Santa Cruz, CAT #8036, USA) and compared to positive and negative controls by flow cytometry. anti-HIS tagsuggested: NoneSoftware and Algorithms Sentences Resources Statistical Analyses: Technical replicates were averaged and data were imported to Prism 8.0 (GraphPad). GraphPadsuggested: (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:There are several limitations to our study that should be noted. First, our cases were all relatively mild (none requiring hospitalization or ventilation), and predominantly middle-aged females. Future studies comparing the responses of older or more severely ill subjects would be informative. Secondly, our study focuses solely on binding of ACE2 to the spike protein and ignores other potential antigens or other immune mechanisms such as inhibition of protease cleavage that might prevent viral entry into the cell. The results of our bead-based assay should be confirmed using a pseudovirus assay, or ideally using live SARS-CoV-2 virus. Third, it is unclear what level of inhibition would correlate with functional resistance to re-infection. Follow-up studies tracking inhibition in our assay over time while simultaneously monitoring subjects for re-infection will be necessary, given the ethical impossibility of experimental human inoculation. Finally, our SARS-CoV-2 PCR+, antibody negative samples leave many questions. For example, did they clear the virus through mechanisms beyond our detection, such as antibodies targeted to alternative viral proteins or non-B-cell dependent mechanisms? Or were their rtPCR results false positives? Most importantly, are they susceptible to future infections? Follow-up with these or similar individuals will be important. In conclusion, the trimer inhibition assay presented here could be broadly useful in the settings of routine clinical evaluati...
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.
-
