ArticleJournal of virological methods2022
An easy pipeline for one-step purification of SARS-CoV-2 nucleocapsid protein from insect cell suspension culture.
Article in Journal of virological methods, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 10 citations in OpenAlex.
- Occlusion body-incorporated SARS-CoV-2 S-RBD via cypovirus polyhedrin-derived peptide: a novel insect cell-expressed antigen for high-accuracy COVID-19 diagnosis, and humoral immune response and hybrid immunity evaluation.Revista do Instituto de Medicina Tropical de Sao Paulo · 2026Article
- Transient Expression in HEK-293 Cells in Suspension Culture as a Rapid and Powerful Tool: SARS-CoV-2 N and Chimeric SARS-CoV-2N-CD154 Proteins as a Case Study.Biomedicines · 2023Article
- Production and Characterization of a SARS-CoV-2 Nucleocapsid Protein Reference Material.ACS measurement science au · 2022Article
- Application of Baculovirus Expression Vector system (BEV) for COVID-19 diagnostics and therapeutics: a review.Journal, genetic engineering & biotechnology · 2022Review
- Understanding the Role of SARS-CoV-2 ORF3a in Viral Pathogenesis and COVID-19.Frontiers in microbiology · 2022Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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.
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