Linear B-cell epitopes in the spike and nucleocapsid proteins as markers of SARS-CoV-2 exposure and disease severity

EBioMedicine. 2020 Aug:58:102911. doi: 10.1016/j.ebiom.2020.102911. Epub 2020 Jul 22.

Abstract

Background: Given the unceasing worldwide surge in COVID-19 cases, there is an imperative need to develop highly specific and sensitive serology assays to define exposure to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).

Methods: Pooled plasma samples from PCR positive COVID-19 patients were used to identify linear B-cell epitopes from a SARS-CoV-2 peptide library of spike (S), envelope (E), membrane (M), and nucleocapsid (N) structural proteins by peptide-based ELISA. Hit epitopes were further validated with 79 COVID-19 patients with different disease severity status, 13 seasonal human CoV, 20 recovered SARS patients and 22 healthy donors.

Findings: Four immunodominant epitopes, S14P5, S20P2, S21P2 and N4P5, were identified on the S and N viral proteins. IgG responses to all identified epitopes displayed a strong detection profile, with N4P5 achieving the highest level of specificity (100%) and sensitivity (>96%) against SARS-CoV-2. Furthermore, the magnitude of IgG responses to S14P5, S21P2 and N4P5 were strongly associated with disease severity.

Interpretation: IgG responses to the peptide epitopes can serve as useful indicators for the degree of immunopathology in COVID-19 patients, and function as higly specific and sensitive sero-immunosurveillance tools for recent or past SARS-CoV-2 infections. The flexibility of these epitopes to be used alone or in combination will allow for the development of improved point-of-care-tests (POCTs).

Funding: Biomedical Research Council (BMRC), the A*ccelerate GAP-funded project (ACCL/19-GAP064-R20H-H) from Agency of Science, Technology and Research (A*STAR), and National Medical Research Council (NMRC) COVID-19 Research fund (COVID19RF-001) and CCGSFPOR20002. ATR is supported by the Singapore International Graduate Award (SINGA), A*STAR.

Keywords: Biomarkers; COVID-19; Epitopes; Patients; SARS-CoV-2.

MeSH terms

  • Adult
  • B-Lymphocytes / immunology*
  • Biomarkers / blood
  • COVID-19
  • Coronavirus Infections / blood
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / immunology
  • Epitopes / blood
  • Epitopes / immunology*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Nucleocapsid Proteins / immunology*
  • Pandemics
  • Pneumonia, Viral / blood
  • Pneumonia, Viral / diagnosis*
  • Pneumonia, Viral / immunology
  • Serologic Tests / methods
  • Spike Glycoprotein, Coronavirus / immunology*

Substances

  • Biomarkers
  • Epitopes
  • Nucleocapsid Proteins
  • Spike Glycoprotein, Coronavirus
  • spike glycoprotein, SARS-CoV