Epitope Mapping on Microarrays Highlights a Sequence on the N Protein with Strong Immune Response in SARS-CoV-2 Patients

Methods Mol Biol. 2023:2578:209-217. doi: 10.1007/978-1-0716-2732-7_15.

Abstract

In SARS-CoV-2 pandemic scenario, the identification of rapid methods to detect antibodies against coronavirus has been a wide and urgent issue. Epitope mapping on peptide microarrays is a rapid way to identify sequences with a high immunoreactivity. The process begins with a proteome-wide screening, based on immune affinity; the use of a high-density microarray is followed by a validation phase, where a restricted panel of probes is tested using peptide microarrays; peptide sequences are immobilized through a click-based strategy.COVID-19-positive sera are tested and immuno-domains regions are identified on SARS-CoV-2 spike (S), nucleocapsid (N) protein, and Orf1ab polyprotein. An epitope on N protein (region 155-171) provided good diagnostic performance in discriminating COVID-19-positive vs. healthy individuals. Using this sequence, 92% sensitivity and 100% specificity are reached for IgG detection in COVID-19 samples, and no cross-reactivity with common cold coronaviruses is detected. Overall, epitope 155-171 from N protein represents a promising candidate for further development and rapid implementation in serological tests.

Keywords: Epitope mapping; Immune-response; Peptide microarrays; SARS-CoV-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Viral
  • COVID-19* / diagnosis
  • Epitope Mapping
  • Epitopes
  • Humans
  • Immunity
  • Immunoglobulin G
  • Polyproteins
  • Proteome
  • SARS-CoV-2* / genetics
  • Spike Glycoprotein, Coronavirus

Substances

  • Antibodies, Viral
  • Epitopes
  • Immunoglobulin G
  • Polyproteins
  • Proteome
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2