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Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2.
bioRxiv. 2020 May 27B

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 EC50 values determined by plaque reduction neutralization test against a clinical isolate of SARS-CoV-2. Taken together, these results demonstrate that the IMMUNO-COV™ assay accurately quantitates SARS-CoV-2 neutralizing antibodies in human sera and therefore is a potentially valuable addition to the currently available serological tests. The assay can provide vital information for comparing immune responses to the various SARS-CoV-2 vaccines that are currently in development, or for evaluating donor eligibility in convalescent plasma therapy studies.

Authors

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Pub Type(s)

Preprint

Language

eng

PubMed ID

32577655

Citation

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, 2020.
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.
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., ... 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 R, et al. Development and Validation of IMMUNO-COV™: a High-throughput Clinical Assay for Detecting Antibodies That Neutralize SARS-CoV-2. bioRxiv. 2020 May 27; PubMed PMID: 32577655.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2. AU - Vandergaast,Rianna, AU - Carey,Timothy, AU - Reiter,Samantha, AU - Lech,Patrycja, AU - Gnanadurai,Clement, AU - Tesfay,Mulu, AU - Buehler,Jason, AU - Suksanpaisan,Lukkana, AU - Naik,Shruthi, AU - Brunton,Bethany, AU - Recker,Jordan, AU - Haselton,Michelle, AU - Ziegler,Christopher, AU - Roesler,Anne, AU - Mills,John R, AU - Theel,Elitza, AU - Weaver,Scott C, AU - Rafael,Grace, AU - Roforth,Matthew M, AU - Jerde,Calvin, AU - Tran,Sheryl, AU - Diaz,Rosa Maria, AU - Bexon,Alice, AU - Baum,Alina, AU - Kyratsous,Christos A, AU - Peng,Kah Whye, AU - Russell,Stephen J, Y1 - 2020/05/27/ PY - 2020/6/25/entrez PY - 2020/6/25/pubmed PY - 2020/6/25/medline JF - bioRxiv : the preprint server for biology JO - bioRxiv N2 - 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 EC50 values determined by plaque reduction neutralization test against a clinical isolate of SARS-CoV-2. Taken together, these results demonstrate that the IMMUNO-COV™ assay accurately quantitates SARS-CoV-2 neutralizing antibodies in human sera and therefore is a potentially valuable addition to the currently available serological tests. The assay can provide vital information for comparing immune responses to the various SARS-CoV-2 vaccines that are currently in development, or for evaluating donor eligibility in convalescent plasma therapy studies. UR - https://www.unboundmedicine.com/medline/citation/32577655/Development_and_validation_of_IMMUNO_COV™:_a_high_throughput_clinical_assay_for_detecting_antibodies_that_neutralize_SARS_CoV_2_ L2 - https://doi.org/10.1101/2020.05.26.117549 DB - PRIME DP - Unbound Medicine ER -
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