Research on the novel coronavirus: SARS-CoV-2 binding assay reagents

basic information

Angiotensin-converting enzyme 2 (ACE2), also known as an ACE homolog, is an endogenous membrane protein that has significant homology with ACE. ACE2 was known to be one of the factors involved in the renin-angiotensin system, which regulates blood pressure. Recently, it has been revealed that the novel coronavirus (SARS-CoV-2), which causes COVID-19, binds to ACE2 present on the cell membrane before being taken up into human cells, suggesting that ACE2 is also a receptor for the novel coronavirus. On our company website (http://www.hakarel.com), we provide detailed binding assay protocols and screening for binding inhibitors using competitive methods on the product introduction page, which can be downloaded and viewed from there. References: (1) Alexandra C. Walls, Young-Jun Park, et al., Cell 180, 1-12 (2020)

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Applications/Examples of results

Human ACE2 protein with His tag (100µg) Usage examples 1. Binding assay The human ACE2 protein with His tag (100µL/well) at concentrations of 0.156-10µg/mL was solid-phase immobilized, and then 0.8µg/mL of biotinylated SARS-CoV-2 spike protein RBD with rabbit Fc tag was bound to it, detected using HRP-conjugated streptavidin. The linear range of the standard curve in this case was 0.156-1.25µg/mL. 2. Screening of binding inhibitors using a competitive method A binding reaction was conducted with solid-phase immobilized human ACE2 protein with His tag (100µL/well) at 1µg/mL and biotinylated SARS-CoV-2 spike protein RBD with rabbit Fc tag at 600ng/mL, to which binding inhibitors (SARS-CoV spike protein RBD with rabbit Fc tag) at concentrations of 10-5000ng/mL were added, and detected using HRP-conjugated streptavidin. The 50% inhibitory concentration (IC50) in this case was 140ng/mL (2.7nM). For other usage examples, please refer to our website.