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

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

in ACE2 / Angiotensin-converting enzyme 2 / Antibodies / Coronaviruses / COVID-19 / Drug development / High-throughput screening / Microbiology / Pandemics / Severe acute respiratory syndrome coronavirus 2 / Spike protein / Trimers

2020

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2020

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2020

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Paper

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

Yuan, Lunzhi,

Yuan, Quan,

Zheng, Qingbing,

Ma, Jian,

Zhang, Tianying,

Hu, Jianda,

Ge, Shengxiang,

Cheng, Tong,

Li, Zonglin,

Ye, Jianghui,

Yang, Shi,

Fu, Baorong,

Wang, Shaojuan,

Hou, Wangheng,

Sheng, Chenguang,

Cao, Jiali,

Zhang, Liang,

Yang, Chuanlai,

Wei, Min,

Xia, Ningshao,

Xiong, Hualong,

Wang, Juan,

Yu, Hai,

Wu, Yangtao,

Liu, Lei,

Li, Zhiyong,

Sheng Nian,

Zhang, Yali,

Xu, Jingjing,

Yang, Ting,

Chen, Yixin,

Zhang, Jun

2020

Overview

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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Publisher

Cold Spring Harbor Laboratory Press,Cold Spring Harbor Laboratory

Subject

ACE2

/ Angiotensin-converting enzyme 2

/ High-throughput screening