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Quantum Dot-Conjugated SARS-CoV-2 Spike Pseudo-Virions Enable Tracking of Angiotensin Converting Enzyme 2 Binding and Endocytosis

Authors
  • Gorshkov, Kirill1
  • Susumu, Kimihiro2, 3
  • Chen, Jiji4
  • Xu, Miao1
  • Pradhan, Manisha1
  • Zhu, Wei1
  • Hu, Xin1
  • Breger, Joyce C.2
  • Wolak, Mason2
  • Oh, Eunkeu2
  • 1 National Center for Advancing Translational Sciences, United States , (United States)
  • 2 Naval Research Laboratory, United States , (United States)
  • 3 Jacobs Corporation, United States , (United States)
  • 4 National Institutes of Health, United States , (United States)
Type
Published Article
Journal
ACS Nano
Publisher
American Chemical Society
Publication Date
Aug 26, 2020
Volume
14
Issue
9
Pages
12234–12247
Identifiers
DOI: 10.1021/acsnano.0c05975
PMID: 32845122
PMCID: PMC7482579
Source
PubMed Central
Keywords
License
Unknown

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.

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