Wang Jonathan Y, Lin Chien-Ju, Liu Ching-Hsuan, Lin Liang-Tzung
Department of Molecular Biosciences, University of Texas at Austin.
School of Pharmacy, College of Pharmacy, Kaohsiung Medical University.
J Vis Exp. 2019 Jul 15(149). doi: 10.3791/59920.
Antiviral assays that mechanistically examine viral entry are pertinent to discern at which step the evaluated agents are most effective, and allow for the identification of candidate viral entry inhibitors. Here, we present the experimental approaches for the identification of small molecules capable of blocking infection by the non-enveloped coxsackievirus A16 (CVA16) through targeting the virus particles or specific steps in early viral entry. Assays include the time-of-drug-addition analysis, flow cytometry-based viral binding assay, and viral inactivation assay. We also present a molecular docking protocol utilizing virus capsid proteins to predict potential residues targeted by the antiviral compounds. These assays should help in the identification of candidate antiviral agents that act on viral entry. Future directions can explore these possible inhibitors for further drug development.
从机制上检查病毒进入的抗病毒检测方法，对于辨别被评估药物在哪个步骤最有效至关重要，并且有助于鉴定候选病毒进入抑制剂。在此，我们展示了通过靶向病毒颗粒或病毒早期进入的特定步骤来鉴定能够阻断无包膜柯萨奇病毒A16（CVA16）感染的小分子的实验方法。检测方法包括药物添加时间分析、基于流式细胞术的病毒结合检测和病毒灭活检测。我们还展示了一种利用病毒衣壳蛋白的分子对接方案，以预测抗病毒化合物靶向的潜在残基。这些检测方法应有助于鉴定作用于病毒进入的候选抗病毒药物。未来的方向可以探索这些可能的抑制剂以进行进一步的药物开发。
