Collaborative Research: A Low-Cost, "Digital" Biosensing Platform with Single Protein Biomarker Sensitivity

蛋白质的准确测量对现代生物医学研究至关重要，特别是对于疾病的早期检测和监测、系统生物学和新药开发。更具体地说，对患者样本中的蛋白质生物标志物进行灵敏、特异、快速和低成本的检测对公共卫生具有重要意义，也是未来个性化诊断和治疗的关键组成部分。到目前为止，所有它们的变体中的酶联免疫吸附分析是用于检测蛋白质生物标记物存在的最常见的传感技术。然而，由于灵敏度低，需要校准图来阐明结果，以及需要先进的昂贵仪器进行超灵敏测量，仍然存在一些限制，极大地限制了它们的广泛应用。这一建议旨在通过开发一种简单、低成本但潜在功能强大的传感器来解决上述缺点，该传感器基于两级信号放大传感机制和机器学习启用的可靠性的集成。开发的传感设备特别适用于疾病的早期检测，理想地能够以良好的便携性、适应性、准确性和高通量“数字”(无需校准)检测生物标记物的存在。从这项研究中获得的知识和技术将传播给科学界和产业界。这个项目将通过将先进的生物传感知识整合到他们的教育和实验室培训中，并积极与普通公众和工业公司互动，来影响研究生、本科生和高中生的教育。所提出的传感设备将允许免疫分析作为一种低成本和数字(无需校准)检测技术进行，通过集成三个关键部件来实现潜在的单拷贝灵敏度：1)压电衬底和可更换的纳米压印声波微柱共振膜之间的耦合共振，2)微米尺寸的质量放大器，通过与检测抗体结合的单分散金纳米颗粒的均匀银增强实现，3)开发和训练机器学习算法，以可靠的方式分析生物传感数据，从而实现更准确的检测。颗粒的数量可以直接与生物标记物的数量相关联，实现“数字化”传感。所开发的方法不限于夹心式免疫分析，也不限于任何特定的蛋白质生物标记物，因此，所开发的传感器可以被推广为监测任何感兴趣的生物分子的通用平台，例如蛋白质生物标记物、病毒和病原体，以确保公共健康和生物安全。如果成功，这项工作将使基于声波的生物传感平台发生革命性变化。这一奖项反映了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。

Accurate measurements of proteins are critical for modern biomedical research, particularly for early detection and monitoring of disease, systems biology, and new drug development. More specifically, sensitive, specific, fast, and low-cost detection of proteins biomarkers in patient samples is of importance for public health and a key component of future personalized diagnostics and therapy. To date, enzyme-linked immune-sorbent assays in all their variants are the most common sensing techniques employed to detect the presence of protein biomarkers. However, a number of limitations still exist that greatly diminish their broader applications due to the low-sensitivity, the requirement of a calibration plot to elucidate the results, and the need for advanced costly instruments for ultrasensitive measurement. This proposal aims to address the aforementioned drawbacks by developing a simple and low-cost, but potentially powerful sensor based on the integration of a two-level signal amplification sensing mechanism and machine-learning-enabled reliability. The developed sensing devices are particularly desirable for early disease detection, ideally capable of "digitally" (calibration-free) detecting the presence of biomarkers with excellent portability, adaptability, accuracy, and high throughput. The knowledge and technology gained from this research will be disseminated to the scientific community and industry. This project will impact the education of the graduate, undergraduate and high school students by integrating advanced biosensing knowledge into their educational and laboratory training, and also actively interacting with general public and industry companies.The proposed sensing device will allow immunoassay to be performed as a low-cost and digital (calibration-free) detection technique with potential single copy sensitivity through integration of three key components: 1) a coupled resonance between a piezoelectric substrate and a replaceable nanoimprinted acoustic wave micropillar resonance film "sticker", 2) a micron-sized mass amplifier realized through uniform silver enhancement of mono-dispersed gold nanoparticles conjugated with detection antibody, and 3) machine learning algorithms to be developed and trained to analyze the biosensing data in a reliable way, thus allowing more accurate detection. The number of the particles can be directly correlated to the number of biomarkers, realizing "digital" sensing. The developed methodology is not restricted to sandwich-type immunoassays, nor is the assay limited to any particular protein biomarker and therefore the develop sensor can be generalized as a universal platform for monitoring any biomolecule of interest such as protein biomarkers, viruses, and pathogens for assuring public health and biosafety. The work will revolutionize the acoustic wave-based biosensing platform if successful.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.