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Development and characterization of a new monoclonal antibody against SARS-CoV-2 NSP12 (RdRp).
Meng, Wen; Guo, Siying; Cao, Simon; Shuda, Masahiro; Robinson-McCarthy, Lindsey R; McCarthy, Kevin R; Shuda, Yoko; Paniz Mondolfi, Alberto E; Bryce, Clare; Grimes, Zachary; Sordillo, Emilia M; Cordon-Cardo, Carlos; Li, Pengfei; Zhang, Hu; Perlman, Stanley; Guo, Haitao; Gao, Shou-Jiang; Chang, Yuan; Moore, Patrick S.
  • Meng W; Cancer Virology Program, University of Pittsburgh Medical Center Hillman Cancer Center, Pittsburgh, Pennsylvania, USA.
  • Guo S; Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Cao S; Cancer Virology Program, University of Pittsburgh Medical Center Hillman Cancer Center, Pittsburgh, Pennsylvania, USA.
  • Shuda M; Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Robinson-McCarthy LR; School of Medicine, Tsinghua University, Beijing, China.
  • McCarthy KR; Cancer Virology Program, University of Pittsburgh Medical Center Hillman Cancer Center, Pittsburgh, Pennsylvania, USA.
  • Shuda Y; Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Paniz Mondolfi AE; Cancer Virology Program, University of Pittsburgh Medical Center Hillman Cancer Center, Pittsburgh, Pennsylvania, USA.
  • Bryce C; Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Grimes Z; Cancer Virology Program, University of Pittsburgh Medical Center Hillman Cancer Center, Pittsburgh, Pennsylvania, USA.
  • Sordillo EM; Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Cordon-Cardo C; Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Li P; Center for Vaccine Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
  • Zhang H; Cancer Virology Program, University of Pittsburgh Medical Center Hillman Cancer Center, Pittsburgh, Pennsylvania, USA.
  • Perlman S; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
  • Guo H; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
  • Gao SJ; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
  • Chang Y; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
  • Moore PS; Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
J Med Virol ; 2022 Oct 21.
Artículo en Inglés | MEDLINE | ID: covidwho-2233227
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.
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Texto completo: Disponible Colección: Bases de datos internacionales Base de datos: MEDLINE Idioma: Inglés Año: 2022 Tipo del documento: Artículo País de afiliación: Jmv.28246

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Texto completo: Disponible Colección: Bases de datos internacionales Base de datos: MEDLINE Idioma: Inglés Año: 2022 Tipo del documento: Artículo País de afiliación: Jmv.28246