Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein

ABSTRACT Immunoassays represent sensitive, easy‐to‐use, and cost‐effective tests useful for the detection of the SARS‐CoV‐2 virus. In this manuscript, we report on the binding specificity of a pair of novel monoclonal antibodies (MAbs) generated against the SARS‐CoV‐2 nucleocapsid protein (NP) and t...

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Main Authors: Robert M. Hnasko, Alice V. Lin, Jeffery A. McGarvey, Eric S. Jackson
Format: Article
Language:English
Published: Wiley 2025-02-01
Series:Microbial Biotechnology
Subjects:
Online Access:https://doi.org/10.1111/1751-7915.70117
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author Robert M. Hnasko
Alice V. Lin
Jeffery A. McGarvey
Eric S. Jackson
author_facet Robert M. Hnasko
Alice V. Lin
Jeffery A. McGarvey
Eric S. Jackson
author_sort Robert M. Hnasko
collection DOAJ
description ABSTRACT Immunoassays represent sensitive, easy‐to‐use, and cost‐effective tests useful for the detection of the SARS‐CoV‐2 virus. In this manuscript, we report on the binding specificity of a pair of novel monoclonal antibodies (MAbs) generated against the SARS‐CoV‐2 nucleocapsid protein (NP) and their development into sensitive sandwich enzyme‐linked immunosorbent assays (sELISA) and a lateral flow immunoassay (LFIA). Binding of these MAbs to hCoVs is limited to variants of SARS‐CoV‐2 and SARS‐CoV NP. Chemiluminescent and absorbance spectroscopy sELISAs report a limit of detection (LOD) for the SARS‐CoV‐2 B.1.1.529 NP variant at 15 pg/mL, and the LFIA using a red‐dyed 200 nm particle at 10 ng/mL. The sELISA exhibits broad SARS‐CoV‐2 viral variant detection with assay LOD for SARS‐CoV‐2 B.1.1.529 virus at 1.4 × 105 genome copies per mL (p ≤ 0.001). The availability of these MAbs should facilitate continued investment in the commercial development of immunoassays to increase global SARS‐CoV‐2 detection technologies.
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spelling doaj-art-82391fd7d32d47e9b82f7620129dfe1f2025-08-20T02:01:58ZengWileyMicrobial Biotechnology1751-79152025-02-01182n/an/a10.1111/1751-7915.70117Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid ProteinRobert M. Hnasko0Alice V. Lin1Jeffery A. McGarvey2Eric S. Jackson3United States Department of Agriculture, Agricultural Research Service Produce Safety and Microbiology Unit Albany California USAUnited States Department of Agriculture, Agricultural Research Service Produce Safety and Microbiology Unit Albany California USAUnited States Department of Agriculture, Agricultural Research Service Foodborne Toxin Detection and Prevention Unit Albany California USAUnited States Department of Agriculture, Agricultural Research Service Foodborne Toxin Detection and Prevention Unit Albany California USAABSTRACT Immunoassays represent sensitive, easy‐to‐use, and cost‐effective tests useful for the detection of the SARS‐CoV‐2 virus. In this manuscript, we report on the binding specificity of a pair of novel monoclonal antibodies (MAbs) generated against the SARS‐CoV‐2 nucleocapsid protein (NP) and their development into sensitive sandwich enzyme‐linked immunosorbent assays (sELISA) and a lateral flow immunoassay (LFIA). Binding of these MAbs to hCoVs is limited to variants of SARS‐CoV‐2 and SARS‐CoV NP. Chemiluminescent and absorbance spectroscopy sELISAs report a limit of detection (LOD) for the SARS‐CoV‐2 B.1.1.529 NP variant at 15 pg/mL, and the LFIA using a red‐dyed 200 nm particle at 10 ng/mL. The sELISA exhibits broad SARS‐CoV‐2 viral variant detection with assay LOD for SARS‐CoV‐2 B.1.1.529 virus at 1.4 × 105 genome copies per mL (p ≤ 0.001). The availability of these MAbs should facilitate continued investment in the commercial development of immunoassays to increase global SARS‐CoV‐2 detection technologies.https://doi.org/10.1111/1751-7915.70117coronavirusdiagnosticELISAimmunoassaylateral flow immunoassaymonoclonal antibodies
spellingShingle Robert M. Hnasko
Alice V. Lin
Jeffery A. McGarvey
Eric S. Jackson
Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein
Microbial Biotechnology
coronavirus
diagnostic
ELISA
immunoassay
lateral flow immunoassay
monoclonal antibodies
title Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein
title_full Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein
title_fullStr Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein
title_full_unstemmed Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein
title_short Immunoassay Detection of SARS‐CoV‐2 Using Monoclonal Antibody Binding to Viral Nucleocapsid Protein
title_sort immunoassay detection of sars cov 2 using monoclonal antibody binding to viral nucleocapsid protein
topic coronavirus
diagnostic
ELISA
immunoassay
lateral flow immunoassay
monoclonal antibodies
url https://doi.org/10.1111/1751-7915.70117
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AT alicevlin immunoassaydetectionofsarscov2usingmonoclonalantibodybindingtoviralnucleocapsidprotein
AT jefferyamcgarvey immunoassaydetectionofsarscov2usingmonoclonalantibodybindingtoviralnucleocapsidprotein
AT ericsjackson immunoassaydetectionofsarscov2usingmonoclonalantibodybindingtoviralnucleocapsidprotein