An easy pipeline for one-step purification of SARS-CoV-2 nucleocapsid protein from insect cell suspension culture

The COVID-19 pandemic has demanded a range of biotechnological products for detection of SARS-CoV-2 variants and evaluation of human seroconversion after infection or vaccination. In this work, we describe an easy pipeline for expression of SARS-CoV-2 nucleocapsid (N) protein in insect cells followe...

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Bibliographic Details
Published in:Journal of virological methods 2022-01, Vol.299, p.114341-114341, Article 114341
Main Authors: de Camargo, Brenda R., da Silva, Leonardo A., de Oliveira, Athos S., Ribeiro, Bergmann M.
Format: Article
Language:English
Subjects:
Affinity chromatography
Animals
Baculovirus
COVID-19
Humans
Insecta
Nucleocapsid
Nucleocapsid protein
Nucleocapsid Proteins - genetics
One-step purification
Pandemics
SARS-CoV-2
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Summary:The COVID-19 pandemic has demanded a range of biotechnological products for detection of SARS-CoV-2 variants and evaluation of human seroconversion after infection or vaccination. In this work, we describe an easy pipeline for expression of SARS-CoV-2 nucleocapsid (N) protein in insect cells followed by its purification via affinity chromatography. The N gene was cloned into the genome of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) via transposition and the resulting recombinant baculovirus was used for infection of lepidopteran Sf9 cells adapted to high-density suspension. Using Tris−HCl pH 8.0 buffer as mobile phase and eluting bound proteins with 175 mM imidazole as part of a three-step gradient, an average of 1 mg N protein could be purified from each 50 mg of total protein from clarified supernatant. Such protein amount allows the manufacturing of serological tests and the development of basic studies on cellular responses to SARS-CoV-2.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2021.114341