Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS-CoV-2 seroconversion

Braz J Microbiol. 2021 Dec;52(4):2069-2073. doi: 10.1007/s42770-021-00556-6. Epub 2021 Aug 3.

Abstract

Serological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here, the SARS-CoV-2 nucleocapsid protein was expressed in Escherichia coli and purified to homogeneity and high yield using a single chromatography step. The purified SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identification of human SARS-CoV-2 seroconverts. The assay sensitivity and specificity were determined analyzing sera from 140 RT-qPCR-confirmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specificity; identical accuracies were obtained in head-to-head comparison with a commercial ELISA kit. Antigen-coated plates were stable for up to 3 months at 4 °C. The ELISA method described is ready for mass production and will be an additional tool to track COVID-19 cases.

Keywords: COVID-19; ELISA; Immunological test; Nucleocapsid protein; SARS-CoV-2.

MeSH terms

  • Antibodies, Viral / blood
  • COVID-19* / diagnosis
  • COVID-19* / immunology
  • Coronavirus Nucleocapsid Proteins / immunology*
  • Enzyme-Linked Immunosorbent Assay*
  • Humans
  • Immunity, Humoral
  • Nucleocapsid Proteins / genetics
  • Phosphoproteins / immunology
  • Sensitivity and Specificity
  • Seroconversion*

Substances

  • Antibodies, Viral
  • Coronavirus Nucleocapsid Proteins
  • Nucleocapsid Proteins
  • Phosphoproteins
  • nucleocapsid phosphoprotein, SARS-CoV-2