A functional assay for serum detection of antibodies against SARS-CoV-2 nucleoprotein

EMBO J. 2021 Sep 1;40(17):e108588. doi: 10.15252/embj.2021108588. Epub 2021 Jul 29.

Abstract

The humoral immune response to SARS-CoV-2 results in antibodies against spike (S) and nucleoprotein (N). However, whilst there are widely available neutralization assays for S antibodies, there is no assay for N-antibody activity. Here, we present a simple in vitro method called EDNA (electroporated-antibody-dependent neutralization assay) that provides a quantitative measure of N-antibody activity in unpurified serum from SARS-CoV-2 convalescents. We show that N antibodies neutralize SARS-CoV-2 intracellularly and cell-autonomously but require the cytosolic Fc receptor TRIM21. Using EDNA, we show that low N-antibody titres can be neutralizing, whilst some convalescents possess serum with high titres but weak activity. N-antibody and N-specific T-cell activity correlates within individuals, suggesting N antibodies may protect against SARS-CoV-2 by promoting antigen presentation. This work highlights the potential benefits of N-based vaccines and provides an in vitro assay to allow the antibodies they induce to be tested.

Keywords: SARS-CoV-2; TRIM21; antibodies; neutralization; nucleoprotein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neutralizing / blood
  • Antibodies, Neutralizing / isolation & purification*
  • Antibodies, Viral / blood
  • Antibodies, Viral / isolation & purification*
  • COVID-19 / blood*
  • COVID-19 / virology
  • Humans
  • Nucleoproteins / blood
  • Nucleoproteins / immunology
  • SARS-CoV-2 / isolation & purification*
  • SARS-CoV-2 / pathogenicity

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Nucleoproteins