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Quantum Dot-Conjugated SARS-CoV-2 Spike Pseudo-Virions Enable Tracking of Angiotensin Converting Enzyme 2 Binding and Endocytosis
- Source :
- ACS Nano; September 2020, Vol. 14 Issue: 9 p12234-12247, 14p
- Publication Year :
- 2020
-
Abstract
- The first step of SARS-CoV-2 infection is binding of the spike protein’s receptor binding domain to the host cell’s ACE2 receptor on the plasma membrane. Here, we have generated a versatile imaging probe using recombinant Spike receptor binding domain conjugated to fluorescent quantum dots (QDs). This probe is capable of engaging in energy transfer quenching with ACE2-conjugated gold nanoparticles to enable monitoring of the binding event in solution. Neutralizing antibodies and recombinant human ACE2 blocked quenching, demonstrating a specific binding interaction. In cells transfected with ACE2-GFP, we observed immediate binding of the probe on the cell surface followed by endocytosis. Neutralizing antibodies and ACE2-Fc fully prevented binding and endocytosis with low nanomolar potency. Importantly, we will be able to use this QD nanoparticle probe to identify and validate inhibitors of the SARS-CoV-2 Spike and ACE2 receptor binding in human cells. This work enables facile, rapid, and high-throughput cell-based screening of inhibitors for coronavirus Spike-mediated cell recognition and entry.
Details
- Language :
- English
- ISSN :
- 19360851 and 1936086X
- Volume :
- 14
- Issue :
- 9
- Database :
- Supplemental Index
- Journal :
- ACS Nano
- Publication Type :
- Periodical
- Accession number :
- ejs54065635
- Full Text :
- https://doi.org/10.1021/acsnano.0c05975