Artículo
Design and optimization of an IgG human ELISA assay reactive to recombinant RBD SARS-CoV-2 protein
Rodríguez, María Celeste
; Ceaglio, Natalia Analia
; Gugliotta, Agustina
; Villarraza, Carlos Javier
; Garay, Ernesto Sergio
; Fuselli, Antonela
; Gastaldi, Victoria
; Tardivo, María Belén; Antuña, Sebastián; Fontana, Diego Sebastian
; Prieto, Claudio
Fecha de publicación:
11/2022
Editorial:
Springer
Revista:
Applied Microbiology and Biotechnology
ISSN:
0175-7598
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Serology assays are essential tools to mitigate the effect of COVID-19, help to identify previous SARS-CoV-2 infections or vaccination, and provide data for surveillance and epidemiologic studies. In this study, we report the production and purification process of the receptor-binding domain (RBD) of SARS-CoV-2 in HEK293 cells, which allowed the design, optimization, and validation of an indirect ELISA (iELISA) for the detection of human anti-RBD antibodies. To find the optimal conditions of this iELISA, a multivariate strategy was performed throughout design of experiments (DoE) and response surface methodology (RSM), one of the main tools of quality by design (QbD) approach. The adoption of this strategy helped to reduce the time and cost during the method development stage and to define an optimum condition within the analyzed design region. The assay was then validated, exhibiting a sensitivity of 94.24 (86.01–98.42%; 95% CI) and a specificity of 95.96% (89.98–98.89%; 95% CI). Besides, the degree of agreement between quality results assessed using kappa’s value was 0.92. Hence, this iELISA represents a high-throughput technique, simple to perform, reliable, and feasible to be scaled up to satisfy the current demands. Since RBD is proposed as the coating antigen, the intended use of this iELISA is not only the detection of previous exposure to the virus, but also the possibility of detecting protective immunity.
Palabras clave:
SARS-COV-2
,
INDIRECT ELISA
,
COVID
,
QbD
,
DoE
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Licencia
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Articulos(SEDE CENTRAL)
Articulos de SEDE CENTRAL
Articulos de SEDE CENTRAL
Citación
Rodríguez, María Celeste; Ceaglio, Natalia Analia; Gugliotta, Agustina; Villarraza, Carlos Javier; Garay, Ernesto Sergio; et al.; Design and optimization of an IgG human ELISA assay reactive to recombinant RBD SARS-CoV-2 protein; Springer; Applied Microbiology and Biotechnology; 106; 23; 11-2022; 7933-7948
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