Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2.
Virology
; 557: 15-22, 2021 05.
Article
em En
| MEDLINE
| ID: mdl-33582454
ABSTRACT
Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production. SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58-419 aa) was expressed in E. coli in soluble form, purified and characterized biochemically and immunologically. Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of ß-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower (94% and 96.5%, respectively). Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.
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MEDLINE
Assunto principal:
Imunoglobulina G
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Ensaio de Imunoadsorção Enzimática
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Proteínas do Nucleocapsídeo de Coronavírus
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SARS-CoV-2
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COVID-19
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Anticorpos Antivirais
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article