Virus-Free and Live-Cell Visualizing SARS-CoV-2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors
Virus-Free and Live-Cell Visualizing SARS-CoV-2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors
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病毒-游离和活细胞可视化 SARS-CoV-2 细胞进入，用于中和抗体和复合抑制剂的研究
DOI：
10.1002/smtd.202001031
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发表时间：
2020-12-18
期刊：
影响因子：
12.4
通讯作者：
Xia, Ningshao
中科院分区：
文献类型：
--
作者：
Zhang, Yali;Wang, Shaojuan;Xia, Ningshao
由SARS-CoV-2感染引起的2019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通过病毒刺突蛋白和ACE 2受体介导进入细胞，是疫苗、治疗性抗体和药物开发的重要靶点。利用哺乳动物细胞表达系统，研制了一种融合荧光蛋白（Gamillus）的SARS-CoV-2刺突三聚体（STG）基因工程传感器，用于检测病毒的侵入过程。在ACE 2表达细胞中，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优异的性能。新系统允许定量分析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了通用工具。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。
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.