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CyDisCo production of functional recombinant SARS-CoV-2 spike receptor binding domain

Protein Sci. 2021-06; 
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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Nucleic Acid Purification & Analysis … Fractions yielding protein (as observed by A280 peaks on the AKTA FPLC (Amersham Biosciences, UK) chromatogram) were gel-verified using a 10% SDS-PAGE gel (Genscript) that was stained with Coomassie Brilliant Blue G-250 for size … Get A Quote

摘要

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 ... More

关键词

COVID19, CyDisCo, SARS-CoV-2, antigen, protein purification