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A simple protein-based surrogate neutralization assay for SARS-CoV-2
Kento T. Abe, … , James M. Rini, Anne-Claude Gingras
Kento T. Abe, … , James M. Rini, Anne-Claude Gingras
Published September 1, 2020
Citation Information: JCI Insight. 2020;5(19):e142362. https://doi.org/10.1172/jci.insight.142362.
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Research Article Infectious disease

A simple protein-based surrogate neutralization assay for SARS-CoV-2

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Abstract

Most of the patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mount a humoral immune response to the virus within a few weeks of infection, but the duration of this response and how it correlates with clinical outcomes has not been completely characterized. Of particular importance is the identification of immune correlates of infection that would support public health decision-making on treatment approaches, vaccination strategies, and convalescent plasma therapy. While ELISA-based assays to detect and quantitate antibodies to SARS-CoV-2 in patient samples have been developed, the detection of neutralizing antibodies typically requires more demanding cell-based viral assays. Here, we present a safe and efficient protein-based assay for the detection of serum and plasma antibodies that block the interaction of the SARS-CoV-2 spike protein receptor binding domain (RBD) with its receptor, angiotensin-converting enzyme 2 (ACE2). The assay serves as a surrogate neutralization assay and is performed on the same platform and in parallel with an ELISA for the detection of antibodies against the RBD, enabling a direct comparison. The results obtained with our assay correlate with those of 2 viral-based assays, a plaque reduction neutralization test (PRNT) that uses live SARS-CoV-2 virus and a spike pseudotyped viral vector–based assay.

Authors

Kento T. Abe, Zhijie Li, Reuben Samson, Payman Samavarchi-Tehrani, Emelissa J. Valcourt, Heidi Wood, Patrick Budylowski, Alan P. Dupuis II, Roxie C. Girardin, Bhavisha Rathod, Jenny H. Wang, Miriam Barrios-Rodiles, Karen Colwill, Allison J. McGeer, Samira Mubareka, Jennifer L. Gommerman, Yves Durocher, Mario Ostrowski, Kathleen A. McDonough, Michael A. Drebot, Steven J. Drews, James M. Rini, Anne-Claude Gingras

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Figure 3

Validation of the snELISA results using orthogonal methods.

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Validation of the snELISA results using orthogonal methods.
(A) Results ...
(A) Results of the plaque reduction neutralization tests on the same samples overlaid on the AUC curves from Figure 2C (n = 57 samples). Color coding indicates the PRNT50 titers, while negative/positive hits on the PRNT90 assay are displayed with a different-sized dot (see Supplemental Figure 11 for additional PRNT results and Supplemental Figure 12 for spike pseudotyped virus results). (B) Correlation between the lentiviral spike pseudotyped virus assay calculated IC50 values and the AUC results from the snELISA. (C and D) Assessment of the ability of monoclonal or affinity reagents to block the interaction between ACE2 and the RBD in the snELISA (C) or the lentiviral spike pseudotyped assay (D) (see Supplemental Figure 13–16 for the direct binding and viral neutralization assays and for additional reagents tested). Values for C were generated from 3 independent experiments, and underlayed points have been placed along the x axis. Values for D were obtained in 1 experiment in technical triplicate. Data are presented as mean ± SEM.

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