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. 2023 May 30;13:8743. doi: 10.1038/s41598-023-35070-2

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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In vitro competitive pseudovirus assay for evaluating the binding affinities of spike variants. Schematic representation of in vitro competitive pseudovirus assay (A). The 293T-hACE2 cells was incubated with D614G spike pseudovirus (expressing luciferase and ZsGreen protein) and different concentration of spike variants. After 60–72 h, infectivity of D614G spike pseudovirus was visualized by expression of ZsGreen protein in fluorescent microscopy at different spike variant treatment (B). The percentage of D614G spike pseudovirus infectivity was quantified by internalized pseudovirus luciferase activity and inhibitory concentration 50 (IC₅₀) of Wild (C) Delta (D) Omicron (E) C.1.2 (F) spike proteins were quantified using non-linear curve fitting models. Table of IC₅₀ and IC₉₀ values of each spike variants (G).