Development and Characterization of a New Monoclonal Antibody against SARS-CoV-2 NSP12

Wen Meng; Siying Guo; Simon Cao; Masahiro Shuda; Lindsey R Robinson-McCarthy; Kevin R McCarthy; Yoko Shuda; Alberto E Paniz Mondolfi; Clare Bryce; Zachary Grimes; Emilia M Sordillo; Carlos Cordon-Cardo; Pengfei Li; Hu Zhang; Stanley Perlman; Haitao Guo; Shou-Jiang Gao; Yuan Chang; Patrick S Moore

doi:10.1002/jmv.28246

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Development and Characterization of a New Monoclonal Antibody against SARS-CoV-2 NSP12

Journal article

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Development and Characterization of a New Monoclonal Antibody against SARS-CoV-2 NSP12

Wen Meng, Siying Guo, Simon Cao, Masahiro Shuda, Lindsey R Robinson-McCarthy, Kevin R McCarthy, Yoko Shuda, Alberto E Paniz Mondolfi, Clare Bryce, Zachary Grimes, …

Journal of medical virology, Vol.95(1), e28246

01/2023

DOI: 10.1002/jmv.28246

PMCID: PMC9874566

PMID: 36271490

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9874566View

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Abstract

SARS-CoV-2 NSP12, the viral RNA-dependent RNA polymerase (RdRp), is required for viral replication and is a therapeutic target to treat COVID-19. To facilitate research on SARS-CoV-2 NSP12 protein, we developed a rat monoclonal antibody (CM12.1) against the NSP12 N-terminus that can facilitate functional studies. Immunoblotting and immunofluorescence assay (IFA) confirmed the specific detection of NSP12 protein by this antibody for cells overexpressing the protein. Although NSP12 is generated from the ORF1ab polyprotein, IFA of human autopsy COVID-19 lung samples revealed NSP12 expression in only a small fraction of lung cells including goblet, club-like, vascular endothelial cells, and a range of immune cells, despite wide-spread tissue expression of spike protein antigen. Similar studies using in vitro infection also generated scant protein detection in cells with established virus replication. These results suggest that NSP12 may have diminished steady-state expression or extensive posttranslation modifications that limit antibody reactivity during SARS-CoV-2 replication.

Details

Title: Subtitle: Development and Characterization of a New Monoclonal Antibody against SARS-CoV-2 NSP12
Creators: Wen Meng
Siying Guo
Simon Cao
Masahiro Shuda
Lindsey R Robinson-McCarthy
Kevin R McCarthy
Yoko Shuda
Alberto E Paniz Mondolfi
Clare Bryce
Zachary Grimes
Emilia M Sordillo
Carlos Cordon-Cardo
Pengfei Li
Hu Zhang
Stanley Perlman
Haitao Guo
Shou-Jiang Gao
Yuan Chang
Patrick S Moore
Resource Type: Journal article
Publication Details: Journal of medical virology, Vol.95(1), e28246
DOI: 10.1002/jmv.28246
PMID: 36271490
PMCID: PMC9874566
ISSN: 0146-6615
eISSN: 1096-9071
Grant note: DOI: 10.13039/100000002, name: National Institutes of Health
Language: English
Electronic publication date: 10/21/2022
Date published: 01/2023
Academic Unit: Microbiology and Immunology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Infectious Disease (Pediatrics)
Record Identifier: 9984306742602771

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