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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| Format: | Article |
| Language: | English |
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Wiley
2025-02-01
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| Series: | Microbial Biotechnology |
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| 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. |
| format | Article |
| id | doaj-art-82391fd7d32d47e9b82f7620129dfe1f |
| institution | OA Journals |
| issn | 1751-7915 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Microbial Biotechnology |
| 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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