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Monoclonal Antibodies Targeting SARS-VOC-2 Nucleocapsid Protein

At a Glance

Researchers at Colorado State University have developed and characterized a panel of monoclonal antibodies targeting the SARS-CoV-2 nucleocapsid protein. The clones were tested for activity in western blots, ELISAs, and immunofluorescence assays with SARS-CoV-2 infected cells and for cross reactivity against SARS-CoV, HuCoV-NL63, and HuCoV-229E N protein. Overall, data provides a strong foundation for the use of the monoclonal antibodies to study SARS-CoV-2 N protein and development of novel diagnostic assays to detection of COVID-19.

Background

Coronavirus nucleocapsid protein plays a key role in coating the viral RNA genome during virion formation, and overall pathogenesis. Additionally, the nucleocapsid protein has been used as a diagnostic marker for active coronavirus infection. There are very few antibody reagents currently available that are specific for SARS-CoV-2 nucleocapsid, as most monoclonal antibodies that are commercially available were generated against SARS-CoV nucleocapsid protein and are cross-reactive for SARS-CoV-2 nucleocapsid. There is a significant need for new antibodies to be produced with increased specificity and utility for detecting SARS-CoV-2 nucleocapsid protein for both the research community and the necessity to develop therapeutics.

Overview

The antibody clones were screened by ELISA, Western Blot, and Immunofluorescence assays. The cross-reactivity of antibodies was tested against SARS-CoV, HuCoV-NL63, and HuCoV-229E N proteins. These antibodies have validated high specificity towards SARS-CoV-2 nucleocapsid protein across several diagnostic and research assays. 

Figure 1: Reactivity of mBG17 and mBG86 Against Captured Virion-Derived N Proteins. Virion-derived N protein serially diluted from 5.5 × 105 PFU/ml stock and captured by anti-NP rabbit polyclonal antibody. mBG17 and mBG86 used in tandem with goat anti-mouse HRP-conjugated antibody to report on mBG effectiveness in capture ELISA.

Benefits

  • Antibodies specifically target SARS-CoV-2 nucleocapsid
  • Nucleocapsid protein is a diagnostic marker for active coronavirus infection
  • Application in ELISA assays, reducing/denatured western blots, immunofluorescence assays, sandwich ELISAs, gene sequences, and others

Applications

  • ELISA
  • Immunofluorescence
  • Immunohistochemistry (IHC)
  • Western blot

Publications

BJ Geiss, et al (2024). PAbFold: Linear Antibody Epitope Prediction using AlphaFold2, https://doi.org/10.7554/eLife.98369.1

JS Terry, et al. (2021) Development of a SARS-COV-2 Nucleocapsid Specific Monoclonal Antibody. Virology. https://doi.org/10.1016/j.virol.2021.01.003.

Last Updated: August 2024
Test tube for COVID-19
Opportunity

Material is available through Addgene, individual product linked below:
211756  – Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17]
211757 – Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86]

Inventors

Brian Geiss
Tony Schountz
James Terry

Reference Number
2020-109
Licensing Manager

Steve Foster
Steve.Foster@colostate.edu
970-491-7100

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