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

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...

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Published in:	Brazilian journal of microbiology Vol. 52; no. 4; pp. 2069 - 2073
Main Authors:	Conzentino, Marcelo S., Forchhammer, Karl, Souza, Emanuel M., Pedrosa, Fábio O., Nogueira, Meri B., Raboni, Sônia M., Rego, Fabiane G. M., Zanette, Dalila L., Aoki, Mateus N., Nardin, Jeanine M., Fornazari, Bruna, Morales, Hugo M. P., Celedon, Paola A. F., Lima, Carla V. P., Mattar, Sibelle B., Lin, Vanessa H., Morello, Luis G., Marchini, Fabricio K., Reis, Rodrigo A., Huergo, Luciano F.
Format:	Journal Article
Language:	English
Published:	Cham Springer International Publishing 01-12-2021 Springer Nature B.V
Subjects:	Antigens Assaying Biomedical and Life Sciences Clinical Microbiology - Short Communication Coronaviruses COVID-19 E coli Enzyme-linked immunosorbent assay Food Microbiology Homogeneity Immune response (humoral) Immunization Life Sciences Mass production Medical Microbiology Microbial Ecology Microbial Genetics and Genomics Microbiology Mycology Nucleocapsids Pandemics Proteins Public health Seroconversion Severe acute respiratory syndrome Severe acute respiratory syndrome coronavirus 2 Vaccines COVID-19 SARS-CoV-2 Nucleocapsid protein Immunological test ELISA
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1517-8382 1678-4405
DOI:	10.1007/s42770-021-00556-6