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Development and characterization of a quantitative ELISA to detect anti-SARS-CoV-2 spike antibodies.

Authors
  • Żak, Magdalena M1
  • Stock, Aryeh2
  • Stadlbauer, Daniel3, 4
  • Zhang, Wei2
  • Cummings, Kirstie5
  • Marsiglia, William6
  • Zargarov, Arsen2
  • Amanat, Fatima3
  • Tamayo, Monica2
  • Cordon-Cardo, Carlos2
  • Krammer, Florian3
  • Mendu, Damodara Rao2
  • 1 Cardiovascular Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY.
  • 2 Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, NY.
  • 3 Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY.
  • 4 Moderna, Inc.
  • 5 Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY.
  • 6 Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY.
Type
Published Article
Journal
Heliyon
Publisher
Elsevier
Publication Date
Nov 20, 2021
Identifiers
DOI: 10.1016/j.heliyon.2021.e08444
PMID: 34841098
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

A novel clinical assay for the detection and quantitation of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was adapted from an in-house, research-based enzyme-linked immunosorbent assay (ELISA). Development and validation were performed under regulatory guidelines, and the test obtained emergency use authorization (EUA) from the New York State Department of Health (NYSDOH) and the Food and Drug Administration (FDA). The Mount Sinai COVID-19 antibody assay is an orthogonal, quantitative direct ELISA test which detects antibodies reactive to the receptor binding domain (RBD) and the spike protein of the novel SARS-CoV-2. The assay is performed on 96-well plates coated with either SARS-CoV-2 recombinant RBD or spike proteins. The test is divided into two stages, a qualitative screening assay against RBD and a quantitative assay against the full-length spike protein. The test uses pooled high titer serum as a reference standard. Negative pre-COVID-19 and positive post-COVID-19, PCR-confirmed specimens were incorporated in each ELISA test run, and the assays were performed independently at two different locations. The Mount Sinai COVID-19 serology performed with high sensitivity and specificity, 92.5% (95% CI: 0.785 - 0.980) and 100% (CI: 0.939 - 1.000) respectively. Between-run precision was assessed with a single run repeated over 22 days; and within-run precision was assessed with 10 replicates per day over 22 days. Both were within reported acceptance criteria (CV ≤20%). This population-based study reveals the applicability and reliability of this novel orthogonal COVID-19 serology test for the detection and quantitation of antibodies against SARS-CoV-2, allowing a broad set of clinical applications, including the broad evaluation of SARS-CoV-2 seroprevalence and antibody profiling in different population subsets. © 2021 Published by Elsevier Ltd.

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