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

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

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1983-1990
Number of pages8
JournalProtein Science
Volume30
Issue number9
DOIs
StatePublished - Sep 2021

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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