ArticleProtein expression and purification2005
Prokaryotic expression, refolding, and purification of fragment 450-650 of the spike protein of SARS-coronavirus.
Article in Protein expression and purification, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 25 citations in OpenAlex.
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- COVID-19 immunotherapy: Treatment based on the immune cell-mediated approaches.International immunopharmacology · 2022Review
- A universal bacteriophage T4 nanoparticle platform to design multiplex SARS-CoV-2 vaccine candidates by CRISPR engineering.Science advances · 2021Article
- Expression of SARS-CoV-2 surface glycoprotein fragment 319-640 in E. coli, and its refolding and purification.Protein expression and purification · 2021Article
- DCs-based therapies: potential strategies in severe SARS-CoV-2 infection.International journal of medical sciences · 2021Review
- Cell adhesion as a novel approach to determining the cellular binding motif on the severe acute respiratory syndrome coronavirus spike protein.Journal of virological methods · 2014Article
- Calreticulin as a hydrophilic chimeric molecular adjuvant enhances IgG responses to the spike protein of severe acute respiratory syndrome coronavirus.Microbiology and immunology · 2012Article
- Comparison of immunoglobulin G responses to the spike and nucleocapsid proteins of severe acute respiratory syndrome (SARS) coronavirus in patients with SARS.Clinical and vaccine immunology : CVI · 2007Article
- Protein folding liquid chromatography and its recent developments.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences · 2007Review
- A study on antigenicity and receptor-binding ability of fragment 450-650 of the spike protein of SARS coronavirus.Virology · 2007Article
- High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells.World journal of gastroenterology · 2006Article
- SARS-CoV genome polymorphism: a bioinformatics study.Genomics, proteomics & bioinformatics · 2005Article
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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 spike (S) glycoprotein is one of the major structure proteins of SARS-associated coronavirus (CoV). Fragment 450-650 (S450-650) of the S protein contains receptor-binding domain and neutralizing epitopes. In this study, S450-650 was expressed with a histidine tag in Escherichia coli BL21. Bacterial inclusion bodies containing the recombinant S450-650 were solubilized with 8 M urea and then applied onto a Ni-nitrilotriacetic acid column. On-column refolding and purification was performed. Reduced glutathione and oxidized glutathione were included in the refolding buffer. In the wash and elution buffers, glycerol and glucose were necessary additives to prevent protein aggregation during purification. This refolding and purification procedure allowed production of S450-650 at up to 500 microg/ml in soluble form, which maintained appropriate antigenicity and immunogenicity. It was able to induce strong IgG responses in BALB/c mice. In Western blot assays, the recombinant S450-650 was recognized by monoclonal Ab against the His-tag and also sera from a convalescent SARS patient. S450-650-based ELISA system was able to detect anti-SARS-CoV IgG Abs in patient sera.
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