Development and characterization of a new monoclonal antibody against SARS-CoV-2 NSP12 (RdRp)

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

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


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.

Original languageEnglish
Article numbere28246
JournalJournal of Medical Virology
Issue number1
StatePublished - Jan 2023


  • COVID-19
  • NSP12
  • RNA-dependent RNA polymerase, RdRp
  • SARS-CoV-2
  • monoclonal antibody


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