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。考虑到蛋白质 - 蛋白质相互作用（PPI）背后的基本原理之一，鉴于许多过程的关键蛋白质在包括但不限于生物体发育，代谢和对病毒的防御方面的关键蛋白质。实验测量蛋白质 - 蛋白质结合力对我们看待和理解生活系统的方式有重大影响。尽管了解分子间结合力的确切值的实用性，但它们的实际测量仍然挑战了蛋白质本质的复杂性。考虑到这一点，我们开发了一种新颖，简单和高通量的微流体平台，以通过轻度操纵对PPI进行超敏化检测。我们制造了Ag/Al涂层的高纵横比纳米柱，用特异性的C反应蛋白和SARS-COV-2尖峰蛋白涂层的表面，后来使用它们分别以等于11和5 pm的限制检测其各自的抗原。我们用聚苯乙烯（PS）薄膜群组装了微流体设备，该薄膜群用PS纳米柱装饰，并在生物素 - 链霉亲素-X系统中检测到PPI，其中X对应于具有各种分子量的配体。优化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。我们认为，这些结果将贡献有关常用蛋白质系统的定量和定性信息，并有助于改善当前可用的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