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Virus-free and live-cell visualizing SARS-CoV-2 cell entry for studies of neutralizing antibodies and compound inhibitors

By Yali Zhang, Shaojuan Wang, Yangtao Wu, Wangheng Hou, Lunzhi Yuan, Chenguang Sheng, Juan Wang, Jianghui Ye, Qingbing Zheng, Jian Ma, Jingjing Xu, Min Wei, Zonglin Li, Sheng Nian, Hualong Xiong, Liang Zhang, Yang Shi, Baorong Fu, Jiali Cao, Chuanlai Yang, Zhiyong Li, Ting Yang, Lei Liu, Hai Yu, Jianda Hu, Shengxiang Ge, Yixin Chen, Tianying Zhang, Jun Zhang, Tong Cheng, Quan Yuan, Ningshao Xia

Posted 22 Jul 2020
bioRxiv DOI: 10.1101/2020.07.22.215236

The ongoing COVID-19 pandemic, caused by SARS-CoV-2 infection, has resulted in hundreds of thousands of deaths. Cellular entry of SARS-CoV-2, which is mediated by the viral spike protein and host ACE2 receptor, is an essential target for the development of vaccines, therapeutic antibodies, and drugs. Using a mammalian cell expression system, we generated a recombinant fluorescent protein (Gamillus)-fused SARS-CoV-2 spike trimer (STG) to probe the viral entry process. In ACE2-expressing cells, we found that the STG probe has excellent performance in the live-cell visualization of receptor binding, cellular uptake, and intracellular trafficking of SARS-CoV-2 under virus-free conditions. The new system allows quantitative analyses of the inhibition potentials and detailed influence of COVID-19-convalescent human plasmas, neutralizing antibodies and compounds, providing a versatile tool for high-throughput screening and phenotypic characterization of SARS-CoV-2 entry inhibitors. This approach may also be adapted to develop a viral entry visualization system for other viruses. ### Competing Interest Statement The authors have declared no competing interest.

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