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Vandergaast R, Carey T, Reiter S, et al. Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2. bioRxiv. 2020;doi: 10.1101/2020.05.26.117549.
Vandergaast, R., Carey, T., Reiter, S., Lech, P., Gnanadurai, C., Tesfay, M., Buehler, J., Suksanpaisan, L., Naik, S., Brunton, B., Recker, J., Haselton, M., Ziegler, C., Roesler, A., Mills, J. R., Theel, E., Weaver, S. C., Rafael, G., Roforth, M. M., Jerde, C., Tran, S., Diaz, R. M., Bexon, A., Baum, A., Kyratsous, C. A., Peng, K. W., & Russell, S. J. (2020). Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2. bioRxiv : the preprint server for biology, . https://doi.org/10.1101/2020.05.26.117549
Vandergaast, Rianna, et al. "Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2." bioRxiv : the preprint server for biology vol. (2020). doi: https://doi.org/10.1101/2020.05.26.117549
Vandergaast R, Carey T, Reiter S, Lech P, Gnanadurai C, Tesfay M, Buehler J, Suksanpaisan L, Naik S, Brunton B, Recker J, Haselton M, Ziegler C, Roesler A, Mills JR, Theel E, Weaver SC, Rafael G, Roforth MM, Jerde C, Tran S, Diaz RM, Bexon A, Baum A, Kyratsous CA, Peng KW, Russell SJ. Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2. bioRxiv. 2020 May 27; doi: 10.1101/2020.05.26.117549. PMID: 32577655; PMCID: PMC7302210.
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bioRxiv. 2020 May 27; doi: 10.1101/2020.05.26.117549.

Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2.

bioRxiv : the preprint server for biology

Rianna Vandergaast, Timothy Carey, Samantha Reiter, Patrycja Lech, Clement Gnanadurai, Mulu Tesfay, Jason Buehler, Lukkana Suksanpaisan, Shruthi Naik, Bethany Brunton, Jordan Recker, Michelle Haselton, Christopher Ziegler, Anne Roesler, John R Mills, Elitza Theel, Scott C Weaver, Grace Rafael, Matthew M Roforth, Calvin Jerde, Sheryl Tran, Rosa Maria Diaz, Alice Bexon, Alina Baum, Christos A Kyratsous, Kah Whye Peng, Stephen J Russell

PMID: 32577655 PMCID: PMC7302210 DOI: 10.1101/2020.05.26.117549

Abstract

We here describe the development and validation of IMMUNO-COV™, a high-throughput clinical test to quantitatively measure SARS-CoV-2-neutralizing antibodies, the specific subset of anti-SARS-CoV-2 antibodies that block viral infection. The test measures the capacity of serum or purified antibodies to neutralize a recombinant Vesicular Stomatitis Virus (VSV) encoding the SARS-CoV-2 spike glycoprotein. This recombinant virus (VSV-SARS-CoV-2-S-Δ19CT) induces fusion in Vero cell monolayers, which is detected as luciferase signal using a dual split protein (DSP) reporter system. VSV-SARS-CoV-2-S-Δ19CT infection was blocked by monoclonal α-SARS-CoV-2-spike antibodies and by plasma or serum from SARS-CoV-2 convalescing individuals. The assay exhibited 100% specificity in validation tests, and across all tests zero false positives were detected. In blinded analyses of 230 serum samples, only two unexpected results were observed based on available clinical data. We observed a perfect correlation between results from our assay and 80 samples that were also assayed using a commercially available ELISA. To quantify the magnitude of the anti-viral response, we generated a calibration curve by adding stepped concentrations of α-SARS-CoV-2-spike monoclonal antibody to pooled SARS-CoV-2 seronegative serum. Using the calibration curve and a single optimal 1:100 serum test dilution, we reliably measured neutralizing antibody levels in each test sample. Virus neutralization units (VNUs) calculated from the assay correlated closely (p < 0.0001) with PRNT

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