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J Immunol Methods ; 495: 113082, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34051226

RESUMO

The development of new diagnostic assays become a priority for managing COVID-19. To this aim, we presented here an in-house ELISA based on the production of two major recombinant and high-quality antigens from SARS-CoV-2. Full-length N and S-RBD fragment proteins fused to mouse IgG2a-Fc were produced in the CHO cell line. Secreted recombinant proteins were easily purified with standard Protein A chromatography and were used in an in-house ELISA to detect anti-N and anti-RBD IgGs in the plasma of COVID-19 RTPCR-positive patients. High reactivity against recombinant antigens was readily detected in all positive plasma samples, whereas no recognition was observed with control healthy subject's plasmas. Remarkably, unpurified recombinant N protein obtained from cell culture supernatant was also suitable for the monitoring by ELISA of IgG levels in positive patients. This work provides an early prospection for low price but high-quality serological kit development.


Assuntos
Teste Sorológico para COVID-19/métodos , COVID-19/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Fragmentos Fc das Imunoglobulinas/metabolismo , Imunoglobulina G/metabolismo , Proteínas Recombinantes/metabolismo , SARS-CoV-2/fisiologia , Animais , Anticorpos Antivirais/sangue , Células CHO , Teste Sorológico para COVID-19/economia , Custos e Análise de Custo , Cricetulus , Humanos , Fragmentos Fc das Imunoglobulinas/genética , Imunoglobulina G/genética , Proteínas Recombinantes/genética , Sensibilidade e Especificidade , Glicoproteína da Espícula de Coronavírus/imunologia
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