CyDisCo production of functional recombinant SARS‐CoV‐2 spike receptor binding domain |
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| Authors: | Janani Prahlad Lucas R. Struble William E. Lutz Savanna A. Wallin Surender Khurana Andy Schnaubelt Mara J. Broadhurst Kenneth W. Bayles Gloria E. O. Borgstahl |
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| Affiliation: | 1. Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha Nebraska, USA ; 2. Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha Nebraska, USA ; 3. Division of Viral Products, Center for Biologics Evaluation and Research (CBER), FDA, Silver Spring Maryland, USA ; 4. Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha Nebraska, USA |
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| Abstract: | ![]()
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. |
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| Keywords: | antigen, COVID19, CyDisCo, protein purification, SARS‐ CoV‐ 2 |
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