Development of Next Generation Plasmonic Nanosensors for Ultrasensitive, High-Throughput Nucleic Acid and Protein Assays

正如SARS-CoV-2大流行所证明的那样，人们越来越需要高度准确和特定的生物传感器来诊断、监测和管理疾病。现有技术通常侧重于单一疾病生物标志物，无法同时对多种生物标志物进行超灵敏检测，最终导致错误的检测结果，特别是在疾病发病时。该项目的目标是通过构建超灵敏的光学生物传感器（称为等离子体纳米传感器）来解决这一限制，该传感器可以通过分析血液和尿液样本来检测标记核酸和蛋白质。此次开发的传感器有可能大幅改善COVID-19和癌症等各种疾病的早期诊断方法。等离子体纳米结构的潜在应用横跨化学和生物化学科学、临床科学和生物工程，以及一般的纳米技术。该项目预计将影响提供多方面的研究和教育方法的努力，通过指导研究和将数据和概念整合到大学课程和现有的社区推广工作中，为下一代创业型科学、技术、工程和数学（STEM）创新者做好准备。本项目的目标是利用局部表面等离子体共振(LSPR)-活性金属纳米结构来设计和构建基于光学的生物传感器，利用简单的紫外-可见吸收分光光度计通过测量分析物附着在生物传感器前后的LSPR峰移来分析不同类别的疾病生物标志物，如核酸和蛋白质。lspr活性金属纳米结构与光开关分子（psm）功能化，作为受体结合基序，允许生物传感器可逆和再生，因此在反复暴露于紫外线和可见光下可重复使用。实验和理论计算相结合的方法可以选择最合适的PSM化学结构和与纳米结构连接的有机配体，以提高生物传感灵敏度。将这种生物传感方法集成到多孔板格式中，可以在一次仪器运行中使用吸收模式的板读取器构建高通量分析，分析数十个患者样品的标准化。这缩短了检测时间，有可能改善临床疾病诊断。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

As evidenced in the SARS-CoV-2 pandemic, there is an ever-increasing need for highly accurate and specific biosensors to diagnose, monitor, and manage illnesses. Existing technologies generally operate with a focus on a single disease biomarker and are unable to perform ultrasensitive assays for multiple biomarkers simultaneously, leading ultimately to false test results, specifically at the disease onset. The goal of this project is to address this limitation by constructing ultrasensitive optical-based biosensors, termed plasmonic nanosensors, that can detect marker nucleic acids and proteins by analyzing blood and urine samples. The sensors developed have the potential to substantially improve the clinical diagnostic approach for early-stage detection of various diseases such as COVID-19 and cancer. The potential applications for plasmonic nanostructures span chemical- and biochemical-sciences, clinical science, and bioengineering, as well as nanotechnology in general. The project is expected to affect efforts to provide multifaceted research and educational approaches to prepare the next generation of entrepreneurial science, technology, engineering, and mathematics (STEM) innovators through mentored-research and the integration of data and concepts into both college coursework and existing community outreach efforts. The goal of this project is to use localized surface plasmon resonance (LSPR)-active metal nanostructures to design and construct optical-based biosensors that can assay different classes of disease biomarkers, such as nucleic acids and proteins, utilizing simple UV-vis absorption spectrophotometer by measuring the LSPR peak shift before and after analyte attachment to biosensors. The LSPR-active metal nanostructures are functionalized with photoswitchable molecules (PSMs) that act as receptor binding motifs allowing the biosensor to be reversible and regenerative, thus reusable, upon repeated exposure to ultraviolet and visible light. A combined experimental and theoretical calculation approach enables selection of the most suitable chemical structure of the PSM and organic ligands connecting it to the nanostructures to enhance the biosensing sensitivity. The integration of this biosensing approach into a multi-well plate format allows construction of a high-throughput assay analyzing a few tens of patient samples with standardization in a single instrument run using a plate reader in the absorption mode. This leads to shorter assay time, with the potential to improve clinical disease diagnosis.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.