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

用于分子间相互作用超灵敏测量的新型微流体平台

• 批准号: 21J10403
• 负责人: Garifullina Ainash
• 金额: $ 0.96万
• 依托单位: Okinawa Institute of Science and Technology Graduate University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-28 至 2023-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21J10403/
• 关键词: Plasmonics; Biosensing; Microfluidics; plasmonics; PPI; SARS-CoV-2; biosensing; LSPR

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.

如今，随着纳米技术的快速发展，我们可以制造设备来回答一些主要的科学问题。这些问题之一涉及蛋白质-蛋白质相互作用 (PPI) 背后的基本原理，因为蛋白质对于许多过程（包括但不限于生物体发育、新陈代谢和防御病毒）至关重要。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具有重大影响。尽管了解分子间结合力的精确值是实用的，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、高通量的微流体平台，用于通过光操纵对 PPI 进行超灵敏检测。我们制造了 Ag/Al 涂层的高纵横比纳米柱，在其表面涂有 C 反应蛋白和 SARS-CoV-2 刺突蛋白特异性抗体，然后用它们检测各自的抗原，检测限分别等于 11 pM 和 5 pM。我们用装饰有 PS 纳米柱的聚苯乙烯 (PS) 薄膜基底组装了微流体装置，并在生物素-链霉亲和素-X 系统中检测了 PPI，其中 X 对应于不同分子量的配体。优化我们的 PPI 检测系统后，我们使用各种蛋白质和生物分子系统收集数据。我们相信这些结果将提供常用蛋白质系统的定量和定性信息，并有助于改进当前可用的 PPI 分析平台。

项目成果

Plasmonic Detection of COVID-19 biomarkers.

COVID-19 生物标志物的等离子检测。

• 发表时间: 2022
• 作者: Ainash Garifullina;Amy Q. Shen;柴 亜東，周 燕ジ，多賀谷 基博;Audisio Tracy Lynn & Bourguignon Thomas;柴 亜東，円子 友理，宮田 真理，多賀谷 基博;Ainash Garifullina
• 通讯作者: Ainash Garifullina

Real-time detection of protein-protein interactions via high aspect ratio plasmonic nanostructures inside a microfluidic channel

通过微流体通道内的高纵横比等离子体纳米结构实时检测蛋白质-蛋白质相互作用

• 发表时间: 2024
• 期刊: Analytical Chemistry
• 影响因子: 7.4
• 作者: Ainash Garifullina;Amy Q. Shen
• 通讯作者: Amy Q. Shen