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Profiling of Surface Protein Epitopes on Viral Particles by Multiplex Dual-Reporter Strategy
Published on: January 12, 2024
Single-Particle Tracking and Positional Phenotyping Reveal Variant-Specific Early Checkpoints in SARS-CoV-2 Cell
Frank H Schulz1,2,3, Marcus W Dreisler1, Denis Koylyu4
1Department of Chemistry & Nanoscience Center, University of Copenhagen, CopenhagenDK-2100, Denmark.
None:
Bulk assays of SARS-CoV-2 entry obscure individual virions and conflate binding with internalization. We developed a quantitative single-particle imaging assay that classifies fluorescent virus-like particles (VLPs) as surface, crossing, or internal on HEK293T-ACE2 cells, separating binding from internalization. Comparing G614 and Omicron BA.5 S protein variants, G614 showed higher binding and a larger internalized fraction at baseline. A trivalent anti-S protein aptamer reduced G614 binding and internalization but increased BA.5 internalization. These positional readouts expose variant-resolved early entry checkpoints and provide a simple platform to test how ligands shift binding and internalization.
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