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Virus‐Free and Live‐Cell Visualizing SARS‐CoV‐2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors
Small Methods ( IF 10.7 ) Pub Date : 2020-12-18 , DOI: 10.1002/smtd.202001031
Yali Zhang 1 , Shaojuan Wang 1 , Yangtao Wu 1 , Wangheng Hou 1 , Lunzhi Yuan 1 , Chenguang Shen 2 , Juan Wang 1 , Jianghui Ye 1 , Qingbing Zheng 1 , Jian Ma 1 , Jingjing Xu 3 , Min Wei 1 , Zonglin Li 1 , Sheng Nian 1 , Hualong Xiong 1 , Liang Zhang 1 , Yang Shi 1 , Baorong Fu 1 , Jiali Cao 1 , Chuanlai Yang 1 , Zhiyong Li 4 , Ting Yang 3 , Lei Liu 2 , Hai Yu 1 , Jianda Hu 3 , Shengxiang Ge 1 , Yixin Chen 1 , Tianying Zhang 1 , Jun Zhang 1 , Tong Cheng 1 , Quan Yuan 1 , Ningshao Xia 1
Affiliation  

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.

中文翻译:

无病毒和活细胞可视化 SARS-CoV-2 细胞进入,用于中和抗体和复合抑制剂的研究

由 SARS-CoV-2 感染引起的 2019 年冠状病毒病(COVID-19)大流行正在持续,已导致数十万人死亡。SARS-CoV-2 的细胞进入由病毒刺突蛋白和 ACE2 受体介导,是开发疫苗、治疗性抗体和药物的重要靶点。使用哺乳动物细胞表达系统,开发了一种融合荧光蛋白(Gamillus)的 SARS-CoV-2 刺突三聚体(STG)的基因工程传感器来探测病毒进入过程。在表达 ACE2 的细胞中,发现 STG 探针在无病毒条件下 SARS-CoV-2 的受体结合、细胞摄取和细胞内运输的活细胞可视化中具有出色的性能。新系统可以定量分析 COVID-19 恢复期人血浆、中和抗体和化合物的抑制潜力和详细影响,为 SARS-CoV-2 进入抑制剂的高通量筛选和表型表征提供通用工具。这种方法也可以适用于开发其他病毒的病毒进入可视化系统。
更新日期:2021-02-12
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