一种用于测量冠状病毒膜融合的无细胞平台。
A cell-free platform to measure coronavirus membrane fusion.
作者信息
Kicmal Thomas, Qing Enya, Hawkins Grant M, Wilcox Alexandria, Gallagher Tom
机构信息
Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USA.
Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL 60153, USA.
出版信息
STAR Protoc. 2023 Mar 6;4(2):102189. doi: 10.1016/j.xpro.2023.102189.
Here we present a protocol to measure coronavirus-mediated membrane fusion, an essential event in coronavirus cell entry. The approach uses nanoluciferase (Nluc) "HiBiT"-tagged corona virus-like particles (VLPs) and Nluc "LgBiT"-containing extracellular vesicles (EVs) as proxies for virus and cell, respectively. VLP-EV membrane fusion allows HiBiT and LgBiT to combine into measurable Nluc, which signifies virus fusion with target cell membranes. We highlight assay utility with methods to assess coronavirus-mediated fusion and its inhibition by antibodies and antiviral agents. For complete details on the use and execution of this protocol, please refer to Qing et al. (2021), Qing et al. (2022), and Marcink et al. (2022)..
在此，我们展示了一种用于测量冠状病毒介导的膜融合的方法，这是冠状病毒进入细胞过程中的一个关键事件。该方法分别使用纳米荧光素酶（Nluc）“HiBiT”标记的冠状病毒样颗粒（VLPs）和含有Nluc“LgBiT”的细胞外囊泡（EVs）作为病毒和细胞的替代物。VLP-EV膜融合使HiBiT和LgBiT结合形成可测量的Nluc，这表明病毒与靶细胞膜发生了融合。我们通过评估冠状病毒介导的融合及其被抗体和抗病毒药物抑制的方法来突出该检测方法的实用性。有关此方法的使用和执行的完整详细信息，请参考Qing等人（2021年）、Qing等人（2022年）以及Marcink等人（2022年）的研究。
Zotero 插件