2020
DOI: 10.1101/2020.05.26.117549
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Development and validation of IMMUNO-COV™: a high-throughput clinical assay for detecting antibodies that neutralize SARS-CoV-2

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 lu… Show more

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Cited by 16 publications
(25 citation statements)
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References 29 publications
(30 reference statements)
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“…Tang 4 Despite widespread interest in neutralizing antibodies, methods for their detection and quantification are relatively low-throughput and limited to Biosafety Level 3-equipped research laboratories. While high-throughput methods have emerged, most rely on recombinant Vesicular Stomatitis Viruses (VSV) engineered to express a portion of the SARS-CoV-2 viral spike protein, and their subsequent entry into cell lines (12)(13)(14).…”
Section: Introductionmentioning
confidence: 99%
“…Tang 4 Despite widespread interest in neutralizing antibodies, methods for their detection and quantification are relatively low-throughput and limited to Biosafety Level 3-equipped research laboratories. While high-throughput methods have emerged, most rely on recombinant Vesicular Stomatitis Viruses (VSV) engineered to express a portion of the SARS-CoV-2 viral spike protein, and their subsequent entry into cell lines (12)(13)(14).…”
Section: Introductionmentioning
confidence: 99%
“…Our previously published SARS-CoV-2 neutralization assay relied upon virus-induced fusion of two dual split protein (DSP) reporter cell lines to generate a luciferase signal (21). To further improve assay throughput and eliminate the need for two cell lines we generated a recombinant VSV (VSV-SARS2-Fluc) encoding SARS-CoV-2 spike-Δ19CT (S-Δ19CT) in place of VSV-G, and firefly luciferase (Fluc) as an additional transcriptional unit located between the S-Δ19CT and VSV-L genes (Fig.…”
Section: Resultsmentioning
confidence: 99%
“…VSV-SARS2-Fluc infects Vero cells via endogenously expressed ACE2 receptors (21). We hypothesized that ACE2 overexpression could enhance Vero cell susceptibility to VSV-SARS2-Fluc and thereby improve assay sensitivity.…”
Section: Resultsmentioning
confidence: 99%
“…Donor blood samples were tested using the Ortho VITROS™ SARS-CoV-2 Total Ig assay, Abbott SARS-CoV-2 IgG assay, in-house ELISAs, and the Vyriad IMMUNO-COV™ neutralization assay as described with some modifications. (23) The IMMUNO-COV assay performed here differed from that which was described in the referenced publication in that: 1) plasma samples were heat-inactivated instead of serum samples, which is necessary due to thermal coagulation and 2) neutralization activity was quantified using neutralization end point titer (NT 100 ) method and not a standard curve.…”
Section: Methodsmentioning
confidence: 99%