A simple and robust approach for expression and purification of Receptor-Binding Domain (RBD) of SARS-CoV-2 spike protein from E. coli
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Updated: Oct 30, 2026
A "Plug-And-Display" Nanoparticle Vaccine Platform Based on Outer Membrane Vesicles Displaying SARS-CoV-2 Receptor-Binding Domain
Published on: July 25, 2022
Solomon Oluwasunmibare Oni1, Shouqi Cui2, Buyuan Ma3
1Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101, China; Shandong Energy Institute, Qingdao, 266101, China; Qingdao New Energy Shandong Laboratory, Qingdao, 266101, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
Abstract:
The Receptor-Binding Domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein is the paramount component that is responsible for recognizing the host receptor angiotensin-converting enzyme 2 (ACE2) to initiate the membrane fusion and cell entry. RBD also serves as an important target for development of neutralizing antibodies, virus inhibitors and vaccines. Here, we developed a one-step purification protocol for getting bacterially overexpressed FLAG tagged-RBD with high yield (>20 mg/L minimal medium) and solubility (∼1 mM), which can be used for Nuclear Magnetic Resonance spectroscopy (NMR) study and structural characterization. The purified RBD retains its biological functions as demonstrated by its binding to the human ACE2 receptor and the neutralizing nanobody Ty1. This protocol provides a simple and robust method for producing RBD samples to facilitate NMR-based studies on SARS-CoV-2 for therapeutic development and understanding viral biology.
