Virus-Free and Live-Cell Visualizing SARS-CoV-2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors.
The ongoing corona virus disease 2019 (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 ACE2 receptor, is an essential target for the development of vaccines, therapeutic antibodies, and drugs. Using a mammalian cell expression system, a genetically engineered sensor of fluorescent protein (Gamillus)-fused SARS-CoV-2 spike trimer (STG) to probe the viral entry process is developed. In ACE2-expressing cells, it is 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.
['Zhang|Yali|Y|', 'Wang|Shaojuan|S|', 'Wu|Yangtao|Y|', 'Hou|Wangheng|W|', 'Yuan|Lunzhi|L|', 'Shen|Chenguang|C|', 'Wang|Juan|J|', 'Ye|Jianghui|J|', 'Zheng|Qingbing|Q|', 'Ma|Jian|J|', 'Xu|Jingjing|J|', 'Wei|Min|M|', 'Li|Zonglin|Z|', 'Nian|Sheng|S|', 'Xiong|Hualong|H|', 'Zhang|Liang|L|', 'Shi|Yang|Y|', 'Fu|Baorong|B|', 'Cao|Jiali|J|', 'Yang|Chuanlai|C|', 'Li|Zhiyong|Z|', 'Yang|Ting|T|', 'Liu|Lei|L|', 'Yu|Hai|H|', 'Hu|Jianda|J|', 'Ge|Shengxiang|S|', 'Chen|Yixin|Y|', 'Zhang|Tianying|T|', 'Zhang|Jun|J|', 'Cheng|Tong|T|', 'Yuan|Quan|Q|', 'Xia|Ningshao|N|https://orcid.org/0000-0003-0179-5266']
fluorescent protein entry inhibitor SARS‐CoV‐2 high‐content analysis live‐cell imaging spike glycoprotein viral entry visualization