ACS Appl Mater Interfaces. 2019 Sep 25;11(38):34586-34594. doi: 10.1021/acsami.9b08521. Epub 2019 Sep 16.
The human immunodeficiency virus 1 (HIV-1) capsid serves as a binding platform for proteins and small molecules from the host cell that regulate various steps in the virus life cycle. However, there are currently no quantitative methods that use assembled capsid lattices to measure host-pathogen interaction dynamics. Here we developed a single-molecule fluorescence biosensor using self-assembled capsid tubes as biorecognition elements and imaged capsid binders using total internal reflection fluorescence microscopy in a microfluidic setup. The method is highly sensitive in its ability to observe and quantify binding, to obtain dissociation constants, and to extract kinetics with an extended application of using more complex analytes that can accelerate characterization of novel capsid binders.
人类免疫缺陷病毒 1（HIV-1）衣壳作为一个结合平台，用于结合宿主细胞中的蛋白质和小分子，调节病毒生命周期的各个步骤。然而，目前还没有使用组装好的衣壳晶格来测量宿主-病原体相互作用动力学的定量方法。在这里，我们开发了一种使用自组装衣壳管作为生物识别元件的单分子荧光生物传感器，并在微流控装置中使用全内反射荧光显微镜对衣壳结合物进行成像。该方法具有很高的灵敏度，能够观察和定量结合，获得解离常数，并提取动力学信息，应用范围广泛，可以使用更复杂的分析物来加速新型衣壳结合物的表征。
