分子間相互作用の超高感度測定のための新しいマイクロ流体プラットフォーム

今天，随着纳米技术的快速发展，我们可以制造设备来回答一些重大的科学问题。其中一个问题涉及蛋白质-蛋白质相互作用(PPI)背后的基本原理，因为蛋白质对许多过程是多么关键，包括但不限于生物体发育、新陈代谢和对病毒的防御。通过实验测量蛋白质-蛋白质结合力对我们如何看待和理解生命系统有着重大影响。尽管知道分子间结合力的精确值是可行的，但考虑到蛋白质性质的复杂性，它们的实际测量仍然具有挑战性。考虑到这一点，我们开发了一种新颖、简单、高通量的微流控平台，用于通过光操纵方式超灵敏地检测PPI。我们制备了银/铝包被的高深宽比纳米管，在其表面包覆了针对C反应蛋白和SARS-CoV-2刺突蛋白的抗体，然后用它们来检测它们各自的抗原，检测限分别为11和5 pm。我们组装了以聚苯乙烯(PS)薄膜为基质的微流控器件，并在生物素-链霉亲和素-X体系中检测到PPI，其中X对应于不同分子量的配体。在优化了我们的PPI检测系统后，我们使用各种蛋白质和生物分子系统收集了数据。我们相信，这些结果将有助于提供有关常用蛋白质系统的定量和定性信息，并有助于改进目前可用的PPI分析平台。

Today, with rapid advancement of nanotechnology, we can fabricate devices to answer some of the major scientific questions. One of these questions deals with fundamentals behind protein-protein interactions (PPI) given how crucial proteins are for many processes including but not limited to organism development, metabolism, and defense against viruses. Experimentally measuring protein-protein binding forces has a significant impact on how we see and understand living systems. Despite the practicality of knowing exact values of intermolecular binding forces, their actual measurement remains challenging given the complexity of proteins’ nature. With this in mind, we developed a novel, simple, and high-throughput microfluidic platform for ultrasensitive detection of PPI by means of light manipulation. We fabricated Ag/Al- coated high aspect ratio nanopillars, coated their surface with antibodies specific to C-reactive protein and SARS-CoV-2 spike protein and later used them to detect their respective antigens with limits of detection equal to 11 and 5 pM, respectively. We assembled microfluidic devices with polystyrene (PS) thin film base decorated with PS nanopillars and detected PPI within Biotin-Streptavidin-X systems, where X corresponds to ligands with various molecular weights. After optimizing our PPI detection system, we collected data using various protein and biomolecule systems. We believe these results will contribute both quantitative and qualitative information on commonly used protein systems and help improve currently available PPI analysis platforms.