Watterson Daniel, Robinson Jodie, Chappell Keith J, Butler Mark S, Edwards David J, Fry Scott R, Bermingham Imogen M, Cooper Matthew A, Young Paul R
School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland 4072, Australia.
Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland 4072, Australia.
Sci Rep. 2016 Mar 15;6:22791. doi: 10.1038/srep22791.
Fusion of the viral envelope with host cell membranes is an essential step in the life cycle of all enveloped viruses. Despite such a clear target for antiviral drug development, few anti-fusion drugs have progressed to market. One significant hurdle is the absence of a generic, high-throughput, reproducible fusion assay. Here we report that real time, label-free measurement of cellular electrical impedance can quantify cell-cell fusion mediated by either individually expressed recombinant viral fusion proteins, or native virus infection. We validated this approach for all three classes of viral fusion and demonstrated utility in quantifying fusion inhibition using antibodies and small molecule inhibitors specific for dengue virus and respiratory syncytial virus.
病毒包膜与宿主细胞膜的融合是所有包膜病毒生命周期中的关键步骤。尽管这是抗病毒药物开发的明确靶点,但很少有抗融合药物进入市场。一个重大障碍是缺乏通用、高通量、可重复的融合检测方法。在此,我们报告细胞电阻抗的实时、无标记测量能够量化由单独表达的重组病毒融合蛋白或天然病毒感染介导的细胞间融合。我们针对所有三类病毒融合验证了该方法,并证明其在使用针对登革热病毒和呼吸道合胞病毒的抗体及小分子抑制剂量化融合抑制方面的实用性。