Artículo
Improved Expression of SARS-CoV-2 Spike RBD Using the Insect Cell-Baculovirus System
Poodts, Joaquín
; Smith, Ignacio
; Birenbaum, Joaquín Manuel
; Rodriguez, María Sol; Montero, Luciano; Wolman, Federico Javier
; Marfía, Juan Ignacio
; Valdez, Silvina Noemi
; Alonso, Leonardo Gabriel
; Targovnik, Alexandra Marisa
; Miranda, Maria Victoria
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Fecha de publicación:
12/2022
Editorial:
MDPI
Revista:
Viruses
ISSN:
1999-4915
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Insect cell-baculovirus expression vector system is one of the most established platforms to produce biological products, and it plays a fundamental role in the context of COVID-19 emergency, providing recombinant proteins for treatment, diagnosis, and prevention. SARS-CoV-2 infection is mediated by the interaction of the spike glycoprotein trimer via its receptor-binding domain (RBD) with the host’s cellular receptor. As RBD is required for many applications, in the context of pandemic it is important to meet the challenge of producing a high amount of recombinant RBD (rRBD). For this reason, in the present study, we developed a process based on Sf9 insect cells to improve rRBD yield. rRBD was recovered from the supernatant of infected cells and easily purified by metal ion affinity chromatography, with a yield of 82% and purity higher than 95%. Expressed under a novel chimeric promoter (polh-pSeL), the yield of rRBD after purification was 21.1 ± 3.7 mg/L, which is the highest performance described in Sf9 cell lines. Finally, rRBD was successfully used in an assay to detect specific antibodies in COVID-19 serum samples. The efficient strategy herein described has the potential to produce high-quality rRBD in Sf9 cell line for diagnostic purpose.
Palabras clave:
BACULOVIRUS
,
COVID-19
,
EXPRESSION
,
INSECT CELLS
,
RBD
,
SARS-COV-2
,
SF9
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Identificadores
Colecciones
Articulos(IDEHU)
Articulos de INST.DE EST.DE LA INMUNIDAD HUMORAL PROF.R.A.MARGNI
Articulos de INST.DE EST.DE LA INMUNIDAD HUMORAL PROF.R.A.MARGNI
Articulos(NANOBIOTEC)
Articulos de INSTITUTO DE NANOBIOTECNOLOGIA
Articulos de INSTITUTO DE NANOBIOTECNOLOGIA
Citación
Poodts, Joaquín; Smith, Ignacio; Birenbaum, Joaquín Manuel; Rodriguez, María Sol; Montero, Luciano; et al.; Improved Expression of SARS-CoV-2 Spike RBD Using the Insect Cell-Baculovirus System; MDPI; Viruses; 14; 12; 12-2022; 1-14
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