Bio-Protocol (May 2021)

Production of the Receptor-binding Domain of the Viral Spike Proteins from 2003 and 2019 SARS CoVs and the Four Common Human Coronaviruses for Serologic Assays and Inhibitor Screening

  • Bruno Segovia-Chumbez,
  • Stephen Graham,
  • Ramesh Jadi,
  • Aravinda de Silva,
  • Lakshmanane Premkumar

DOI
https://doi.org/10.21769/BioProtoc.4026
Journal volume & issue
Vol. 11, no. 10

Abstract

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The recombinant receptor-binding domain (RBD) of the viral spike protein from SARS-CoV-1 and 2 are reliable antigens for detecting viral-specific antibodies in humans. We and others have shown that the levels of RBD-binding antibodies and SARS-CoV-2 neutralizing antibodies in patients are correlated. Here, we report the expression and purification of properly folded RBD proteins from SARS and common-cold HCoVs in mammalian cells. RBD proteins were produced with cleavable tags for affinity purification from the cell culture medium and to support multiple immunoassay platforms and drug discovery efforts.Graphic abstract:High-Yield Production of Viral Spike RBDs for Diagnostics and Drug Discovery