Author | Conzentino, Marcelo S. | |
Author | Gonçalves, Ana C. A. | |
Author | Paula, Nigella M. | |
Author | Rego, Fabiane G. M. | |
Author | Zanette, Dalila Lucíola | |
Author | Aoki, Mateus Nóbrega | |
Author | Nardin, Jeanine M. | |
Author | Huergo, Luciano Fernandes | |
Access date | 2022-05-25T18:59:15Z | |
Available date | 2022-05-25T18:59:15Z | |
Document date | 2022 | |
Citation | CONZENTINO, Marcelo S. et al. A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS‑CoV‑2 Spike S1 RBD produced in Escherichia coli. Brazilian Journal of Microbiology, p. 1–7, 2022. | pt_BR |
ISSN | 1678-4405 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/52882 | |
Language | por | pt_BR |
Publisher | Sociedade Brasileira de Microbiologia | pt_BR |
Rights | open access | pt_BR |
Subject in Portuguese | COVID-19 | pt_BR |
Subject in Portuguese | SARS-CoV-2 | pt_BR |
Subject in Portuguese | Magnetic beads | pt_BR |
Title | A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS‑CoV‑2 Spike S1 RBD produced in Escherichia coli | pt_BR |
Type | Article | pt_BR |
DOI | https://doi.org/10.1007/s42770-022-00753-x | |
Abstract | Immunological assays to detect SARS-CoV-2 Spike Receptor Binding Domain (RBD) antigen seroconversion in humans are important tools to monitor the levels of protecting antibodies in the population in response to infection and/or immunization. Here we describe a simple, low cost, and high throughput Ni2+ magnetic bead immunoassay to detect human IgG reactive to Spike S1 RBD Receptor Binding Domain produced in Escherichia coli. A 6xHis-tagged Spike S1 RBD was expressed in E. coli and purifed by afnity chromatography. The protein was mobilized on the surface of Ni2+ magnetic beads and used to investigate the presence of reactive IgG in the serum obtained from pre-pandemic and COVID-19 confrmed cases. The method was validated with a cohort of 290 samples and an area under the receiver operating characteristic curve of 0.94 was obtained. The method was operated with>82% sensitivity at 98% specifcity and was also able to track human IgG raised in response to vaccination with Comirnaty at>85% sensitivity. The IgG signal obtained with the described method was well-correlated with the signal obtained when pre fusion Spike produced in HEK cell lines was used as antigen. This novel low-cost and high throughput immunoassay may act as an important tool to investigate protecting IgG antibodies against SARS-CoV-2 in the human population. | pt_BR |
Affilliation | Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil. | pt_BR |
Affilliation | Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil. | pt_BR |
Affilliation | Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil. | pt_BR |
Affilliation | Universidade Federal do Paraná. Programa de Pós-Graduação em Ciências Farmacêuticas. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Hospital Erasto Gaertner. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil. | pt_BR |
Subject | Immunoassay | pt_BR |
Subject | Magnetics | pt_BR |
Subject | Receptors, Coronavirus | pt_BR |
Subject | Antibodies | pt_BR |
Subject | Receptors, IgG | pt_BR |
Subject in Spanish | Inmunoensayo | pt_BR |
Subject in Spanish | Receptores de Coronavirus | pt_BR |
Subject in Spanish | Anticuerpos | pt_BR |
Subject in French | Dosage immunologique | pt_BR |
Subject in French | Magnétisme | pt_BR |
Subject in French | Anticorps | pt_BR |
Subject in French | Récepteurs du fragment Fc des IgG | pt_BR |
DeCS | Imunoensaio | pt_BR |
DeCS | Magnetismo | pt_BR |
DeCS | Receptores de Coronavírus | pt_BR |
DeCS | Anticorpos | pt_BR |
DeCS | Receptores de IgG | pt_BR |