Bacterial expression and purification of functional recombinant SARS-CoV-2 spike receptor binding domain

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 the SARS-CoV-2 spike protein re...

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Bibliographic Details
Published inbioRxiv
Main Authors Janani Prahlad, Struble, Lucas R, Lutz, William E, Wallin, Savanna A, Khurana, Surender, Schnaubelt, Andy, Broadhurst, Mara J, Bayles, Kenneth W, Borgstahl, Gloria E O
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 03.02.2021
Subjects
ACE2
Angiotensin
Angiotensin-converting enzyme 2
Coronaviruses
COVID-19
Disulfide bonds
Pandemics
Peptidyl-dipeptidase A
Protein purification
Severe acute respiratory syndrome coronavirus 2
Spike protein
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Summary: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 the SARS-CoV-2 spike protein receptor binding domain using the CyDisCo system to create and maintain the correct disulfide bonds for protein integrity and functionality. We show that it is possible to quickly and inexpensively produce functional, active antigen in bacteria capable of recognizing and binding to the ACE2 (angiotensin-converting enzyme) receptor as well as antibodies in COVID-19 patient sera.
DOI:10.1101/2021.02.03.429601