Highly specific monoclonal antibodies and epitope identification against SARS-CoV-2 nucleocapsid protein for antigen detection tests

Cell Rep Med. 2021 Jun 15;2(6):100311. doi: 10.1016/j.xcrm.2021.100311. Epub 2021 May 16.

Abstract

The ongoing coronavirus disease 2019 (COVID-19) pandemic is a major global public health concern. Although rapid point-of-care testing for detecting viral antigen is important for management of the outbreak, the current antigen tests are less sensitive than nucleic acid testing. In our current study, we produce monoclonal antibodies (mAbs) that exclusively react with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and exhibit no cross-reactivity with other human coronaviruses, including SARS-CoV. Molecular modeling suggests that the mAbs bind to epitopes present on the exterior surface of the nucleocapsid, making them suitable for detecting SARS-CoV-2 in clinical samples. We further select the optimal pair of anti-SARS-CoV-2 nucleocapsid protein (NP) mAbs using ELISA and then use this mAb pair to develop immunochromatographic assay augmented with silver amplification technology. Our mAbs recognize the variants of concern (501Y.V1-V3) that are currently in circulation. Because of their high performance, the mAbs of this study can serve as good candidates for developing antigen detection kits for COVID-19.

Keywords: COVID-19; SARS-CoV-2; monoclonal antibody; nucleoprotein; point-of-care testing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antigen-Antibody Reactions
  • COVID-19 / pathology
  • COVID-19 / virology
  • Coronavirus Nucleocapsid Proteins / genetics
  • Coronavirus Nucleocapsid Proteins / immunology*
  • Coronavirus Nucleocapsid Proteins / metabolism
  • Epitopes / immunology*
  • Humans
  • Immunization
  • Immunoassay / methods*
  • Mice
  • Mice, Inbred BALB C
  • Phosphoproteins / genetics
  • Phosphoproteins / immunology
  • Phosphoproteins / metabolism
  • Point-of-Care Systems
  • SARS-CoV-2 / isolation & purification
  • SARS-CoV-2 / metabolism*
  • Silver / chemistry

Substances

  • Antibodies, Monoclonal
  • Coronavirus Nucleocapsid Proteins
  • Epitopes
  • Phosphoproteins
  • nucleocapsid phosphoprotein, SARS-CoV-2
  • Silver