CyDisCo production of functional recombinant SARS‐CoV ‐2 spike receptor binding domain
| Autor: | Andy Schnaubelt, William E. Lutz, Gloria E. O. Borgstahl, Janani Prahlad, Mara J. Broadhurst, Kenneth W. Bayles, Surender Khurana, Savanna A. Wallin, Lucas R. Struble |
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| Rok vydání: | 2021 |
| Předmět: |
Methods and Applications
COVID19 Recombinant Fusion Proteins CyDisCo Biochemistry SARS‐CoV‐2 law.invention antigen Protein Domains law protein purification Escherichia coli Humans Protein disulfide-isomerase Receptor Molecular Biology chemistry.chemical_classification biology SARS-CoV-2 Chemistry biology.organism_classification Folding (chemistry) Enzyme Cytoplasm Spike Glycoprotein Coronavirus biology.protein Recombinant DNA Angiotensin-Converting Enzyme 2 Antibody Bacteria |
| Zdroj: | Protein Science : A Publication of the Protein Society |
| ISSN: | 1469-896X 0961-8368 |
| Popis: | The COVID‐19 pandemic caused by SARS‐CoV‐2 has applied significant pressure on overtaxed healthcare around the world, underscoring the urgent need for rapid diagnosis and treatment. We have developed a bacterial strategy for the expression and purification of a SARS‐CoV‐2 spike protein receptor binding domain (RBD) that includes the SD1 domain. Bacterial cytoplasm is a reductive environment, which is problematic when the recombinant protein of interest requires complicated folding and/or processing. The use of the CyDisCo system (cytoplasmic disulfide bond formation in E. coli) bypasses this issue by pre‐expressing a sulfhydryl oxidase and a disulfide isomerase, allowing the recombinant protein to be correctly folded with disulfide bonds for protein integrity and functionality. We show that it is possible to quickly and inexpensively produce an active RBD in bacteria that is capable of recognizing and binding to the ACE2 (angiotensin‐converting enzyme) receptor as well as antibodies in COVID‐19 patient sera. |
| Databáze: | OpenAIRE |
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