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用于严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染过程和生命周期中关键病毒蛋白的基于荧光的检测方法的开发。

Development of Fluorescence-Based Assays for Key Viral Proteins in the SARS-CoV-2 Infection Process and Lifecycle.

作者信息

Deng Mingzhenlong, Zhang Chuang, Yan Wanli, Chen Lei, He Bin, Li Yan

机构信息

State Key Laboratory of Functions and Applications of Medicinal Plants, Engineering Research Center for the Development and Application of Ethnic Medicine and TCM (Ministry of Education), Guizhou Provincial Key Laboratory of Pharmaceutics, School of Pharmacy, Guizhou Medical University, Guiyang 550004, China.

School of Basic Medical Science, Guizhou Medical University, Guiyang 550004, China.

出版信息

Int J Mol Sci. 2024 Mar 1;25(5):2850. doi: 10.3390/ijms25052850.

DOI:10.3390/ijms25052850
PMID:38474097
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10932265/
Abstract

Since the appearance of SARS-CoV-2 in 2019, the ensuing COVID-19 (Corona Virus Disease 2019) pandemic has posed a significant threat to the global public health system, human health, life, and economic well-being. Researchers worldwide have devoted considerable efforts to curb its spread and development. The latest studies have identified five viral proteins, spike protein (Spike), viral main protease (3CLpro), papain-like protease (PLpro), RNA-dependent RNA polymerase (RdRp), and viral helicase (Helicase), which play crucial roles in the invasion of SARS-CoV-2 into the human body and its lifecycle. The development of novel anti-SARS-CoV-2 drugs targeting these five viral proteins holds immense promise. Therefore, the development of efficient, high-throughput screening methodologies specifically designed for these viral proteins is of utmost importance. Currently, a plethora of screening techniques exists, with fluorescence-based assays emerging as predominant contenders. In this review, we elucidate the foundational principles and methodologies underpinning fluorescence-based screening approaches directed at these pivotal viral targets, hoping to guide researchers in the judicious selection and refinement of screening strategies, thereby facilitating the discovery and development of lead compounds for anti-SARS-CoV-2 pharmaceuticals.

摘要

自2019年严重急性呼吸综合征冠状病毒2（SARS-CoV-2）出现以来，随之而来的2019冠状病毒病（COVID-19）大流行对全球公共卫生系统、人类健康、生命和经济福祉构成了重大威胁。世界各地的研究人员为遏制其传播和发展付出了巨大努力。最新研究确定了五种病毒蛋白，即刺突蛋白（Spike）、病毒主蛋白酶（3CLpro）、木瓜样蛋白酶（PLpro）、RNA依赖性RNA聚合酶（RdRp）和病毒解旋酶（Helicase），它们在SARS-CoV-2侵入人体及其生命周期中发挥着关键作用。针对这五种病毒蛋白开发新型抗SARS-CoV-2药物具有巨大潜力。因此，开发专门针对这些病毒蛋白的高效、高通量筛选方法至关重要。目前，存在大量筛选技术，基于荧光的检测方法成为主要的竞争方法。在本综述中，我们阐明了针对这些关键病毒靶点的基于荧光的筛选方法的基本原理和方法，希望指导研究人员明智地选择和优化筛选策略，从而促进抗SARS-CoV-2药物先导化合物的发现和开发。

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