Institute of Virology, University of Veterinary Medicine, Vienna, Veterinarplatz 1, 1210 Vienna, Austria.
Mol Biotechnol. 2013 Jan;53(1):9-18. doi: 10.1007/s12033-012-9616-6.
In this study, we describe a versatile, flexible, and quick method to label different families of enveloped viruses with glycosylphosphatidylinositol-modified green fluorescent protein, termed fluorescence molecular painting (FMP). As an example for a potential application, we investigated virus attachment by means of flow cytometry to determine if viral binding behavior may be analyzed after FMP of enveloped viruses. Virus attachment was inhibited by using either dextran sulfate or by blocking attachment sites with virus pre-treatment. Results from the FMP-flow cytometry approach were verified by immunoblotting and enzyme-linked immunosorbent assay. Since the modification strategy is applicable to a broad range of proteins and viruses, variations of this method may be useful in a range of research and applied applications from bio-distribution studies to vaccine development and targeted infection for gene delivery.
在这项研究中，我们描述了一种通用、灵活和快速的方法，即用糖基磷脂酰肌醇修饰的绿色荧光蛋白对不同家族的包膜病毒进行标记，称为荧光分子作图（FMP）。作为潜在应用的一个例子，我们通过流式细胞术研究了病毒的附着，以确定 FMP 处理后的包膜病毒是否可以分析病毒的结合行为。通过使用硫酸葡聚糖或通过用病毒预处理来阻断附着位点，抑制了病毒附着。FMP-流式细胞术方法的结果通过免疫印迹和酶联免疫吸附试验得到验证。由于该修饰策略适用于广泛的蛋白质和病毒，因此该方法的变体可能在从生物分布研究到疫苗开发和靶向感染基因传递等一系列研究和应用中具有用途。
