Competitive SARS-CoV-2 Serology Reveals Most Antibodies Targeting the Spike Receptor-Binding Domain Compete for ACE2 Binding
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Abstract
With the emergence and continued spread of the SARS-CoV-2 virus, and of the associated disease, coronavirus disease 2019 (COVID-19), there is an urgent need for improved understanding of how the body mounts an immune response to the virus. Here, we developed a competitive SARS-CoV-2 serological assay that can simultaneously determine whether an individual has developed antibodies against the SARS-CoV-2 Spike protein receptor-binding domain (RBD) and measure the proportion of these antibodies that block interaction with the human angiotensin-converting enzyme 2 (ACE2) required for viral entry. Using this assay and 144 SARS-CoV-2 patient serum samples, we found that a majority of anti-RBD antibodies compete for ACE2 binding. These results not only highlight the need to design vaccines to generate such blocking antibodies but also demonstrate the utility of this assay to rapidly screen patient sera for potentially neutralizing antibodies.
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SciScore for 10.1101/2020.05.27.20114652: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Patient Samples: All serum samples were obtained via antecubital venipuncture and collected into BD Vacutainer serum collection tubes in using protocols approved by the UCSF Institutional Review Board and in accordance with the Declaration of Helsinki. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Plates were again washed, and antibodies bound to the coated antigens detected using 20 μL of anti-human Fab (Jackson ImmunoResearch Laboratories 109-036-097, [1:5000]), anti-human IgM (Sigma-Aldrich A6907, [1:3000]), anti-human IgG (Sigma-Aldrich … SciScore for 10.1101/2020.05.27.20114652: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Patient Samples: All serum samples were obtained via antecubital venipuncture and collected into BD Vacutainer serum collection tubes in using protocols approved by the UCSF Institutional Review Board and in accordance with the Declaration of Helsinki. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Plates were again washed, and antibodies bound to the coated antigens detected using 20 μL of anti-human Fab (Jackson ImmunoResearch Laboratories 109-036-097, [1:5000]), anti-human IgM (Sigma-Aldrich A6907, [1:3000]), anti-human IgG (Sigma-Aldrich A0170, [1:3000]), or Protein L (Thermo Fisher Scientific 32420, [1:5000]) as indicated for 30 minutes at room temperature. anti-human Fabsuggested: Noneanti-human IgMsuggested: (Sigma-Aldrich Cat# A6907, RRID:AB_258318)anti-human IgGsuggested: (Sigma-Aldrich Cat# A0170, RRID:AB_257868)Software and Algorithms Sentences Resources All graphing and statistical analysis was performed in GraphPad Prism (Version 8.4.2). GraphPad Prismsuggested: (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.
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