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，对用于诊断、监测和管理疾病的高度准确和特定的生物传感器的需求日益增长。现有技术通常专注于单一疾病生物标记物，无法同时对多个生物标记物进行超灵敏分析，最终导致错误的测试结果，特别是在疾病开始时。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，通过分析血液和尿液样本来检测标志性核酸和蛋白质，以解决这一限制。开发的传感器有可能大幅改进对各种疾病的早期检测的临床诊断方法，如新冠肺炎和癌症。等离子体纳米结构的潜在应用范围包括化学和生化科学、临床科学、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研究和教育方法的努力，以通过指导研究以及将数据和概念整合到大学课程和现有的社区推广工作中来培养下一代创业型科学、技术、工程和数学(STEM)创新者。本项目的目标是利用局域表面等离子体共振(LSPR)活性金属纳米结构来设计和构建光学生物传感器，通过测量分析物与生物传感器连接前后的LSPR峰位移，利用简单的UV-Vis吸收光谱仪来检测不同类型的疾病生物标志物，如核酸和蛋白质。LSPR活性的金属纳米结构被光开关分子(PSM)功能化，这些PSM作为受体结合基序，使得生物传感器在反复暴露于紫外线和可见光时是可逆的和可再生的，从而可以重复使用。实验和理论计算相结合的方法可以选择最合适的PSM的化学结构和将其连接到纳米结构的有机配体，以提高生物传感的灵敏度。将这种生物传感方法集成到多孔平板格式中，可以构建高通量分析方法，通过在吸收模式下使用平板读取器在单个仪器上运行标准化来分析数十个患者样本。这将缩短化验时间，并有可能改善临床疾病诊断。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.