Kipper Sarit, Avrahami Dorit, Bajorek Monika, Gerber Doron
Nanotechnology Institute, Mina and Everard Goodman Faculty of Life Sciences, Bar Ilan University, Ramat Gan, Israel.
Section of Virology, Faculty of Medicine, Imperial College London, St. Mary's Campus, Norfolk Place, London, W2 UK, UK.
Methods Mol Biol. 2016;1442:165-74. doi: 10.1007/978-1-4939-3687-8_12.
We present a high-throughput microfluidics platform to identify novel host cell binding partners of respiratory syncytial virus (RSV) matrix (M) protein. The device consists of thousands of reaction chambers controlled by micro-mechanical valves. The microfluidic device is mated to a microarray-printed custom-made gene library. These genes are then transcribed and translated on-chip, resulting in a protein array ready for binding to RSV M protein.Even small viral proteome, such as that of RSV, presents a challenge due to the fact that viral proteins are usually multifunctional and thus their interaction with the host is complex. Protein microarrays technology allows the interrogation of protein-protein interactions, which could possibly overcome obstacles by using conventional high throughput methods. Using microfluidics platform we have identified new host interactors of M involved in various cellular pathways. A number of microfluidics based assays have already provided novel insights into the virus-host interactome, and the results have important implications for future antiviral strategies aimed at targets of viral protein interactions with the host.
我们展示了一个高通量微流控平台,用于鉴定呼吸道合胞病毒(RSV)基质(M)蛋白新的宿主细胞结合伴侣。该设备由数千个由微机械阀控制的反应室组成。微流控设备与微阵列打印的定制基因文库相结合。然后这些基因在芯片上进行转录和翻译,形成一个可供与RSV M蛋白结合的蛋白质阵列。即使是像RSV这样的小型病毒蛋白质组,由于病毒蛋白通常具有多种功能,因此它们与宿主的相互作用很复杂,这也带来了挑战。蛋白质微阵列技术允许对蛋白质-蛋白质相互作用进行研究,这可能通过使用传统的高通量方法克服障碍。利用微流控平台,我们已经鉴定出参与各种细胞途径的M蛋白新的宿主相互作用分子。许多基于微流控的检测方法已经为病毒-宿主相互作用组提供了新的见解,这些结果对于未来针对病毒蛋白与宿主相互作用靶点的抗病毒策略具有重要意义。