CAREER: Nanoelectronic Microfluidic Biochip for Ultrasensitive Detection of Selective Protein Biomarkers

本提案的目的是通过在纳米/微流体通道/腔室中使用多路纳米级电极和芯片上数据处理的适体-蛋白质相互作用，开发分离和检测选择性蛋白质（生物标志物）的新模式。为了实现目标，将采用连贯的制造和建模策略：(1)开发分离低丰度疾病生物标志物的功能化膜；(2)设计和开发具有可单独寻址的纳米电极的生物芯片，采用高通量纳米压印光刻技术制成，并使用适配体进行功能化，用于生物标志物的多路检测；(3)开发新型和快速制造纳米/微流体通道；(4)建模；(5)实时低功耗无噪声读出电路，对识别位点进行顺序寻址、驱动、测量和数据分析。知识优势：该提案将改变并创造一个新的领域——蛋白质学？在蛋白质组学的进步的基础上，然后呢?分子电子学吗?。这些活动利用了分子尺度装置和体外适体-蛋白质相互作用，并可扩展到许多其他应用。这些想法将克服分子电子学昂贵和连续制造的瓶颈，并为劳动密集型、低灵敏度和冗长的蛋白质组学协议提供替代方案。新型聚合物纳米/微流体将为保持蛋白质表达和功能提供适当的条件。芯片上的电路将会成为即时护理蛋白质生物芯片的原型。纳米电极将为适配体-蛋白质结合提供三维相互作用体积，从而获得比平面电极更高的灵敏度和信噪比。该方法还将通过消除器件掺杂、几何形状、尺寸和流体环境的影响来克服灵敏度限制。所提出的策略将创新性地改变和革新许多学科：(1)用于生物传感的快速纳米制造，(2)使用各种适体对疾病标记物进行多路检测，(3)用于早期疾病检测的生物标记物的超灵敏芯片上电检测和分析，(4)新型纳米/微流体的无口罩生产。更广泛的影响：该提案在其他生物传感器领域有直接应用，例如基因表达分析，病毒/病原体检测和全血分析。提出的技术的变化可以改变生物分子传感与更好的疾病干预策略，提高统计置信度和实时检测。PI已经聘请了女性研究生和少数民族本科生/高中生在他的研究实验室工作。本建议将继续进行创新教育工作：(1)开发纳米生物器件的研究生课程，(2)研讨会/演示/实验室参观，重点关注本科生的研究参与和保留，(3)为来自阿灵顿学区的高中生（主要是非裔美国人和西班牙裔）提供为期一周的夏令营，整合MEMS/纳米研究和生物学概念，(4)交互式网站/博客，用于投影/展示/讨论研究中的最新技术。(5)周六上午的实时聊天，以跟进/参与K-12学生和教师；(6)技术转移研究，以培养对现实问题感兴趣的学生的创业精神；(7)发展国际研究合作，以交换来自美国和美国的学生。拟议想法的结果将通过同行评议的文章、会议和公共媒体传播。

The objective of this proposal is to develop new modalities for the isolation and detection of selective proteins (biomarkers), by using aptamer-protein interactions in nano/microfluidic channels/chambers with multiplexed nanoscale electrodes and on-chip data processing. To accomplish the goals, a coherent strategy of fabrication and modeling will be adopted: (1) Development of functionalized membranes for isolation of low-abundant disease biomarkers, (2) Design and development of a bio-chip with individually-addressable nano-electrodes, made with high-throughput nanoimprint lithography and functionalized with aptamers for multiplexed detection of biomarkers, (3) Development of novel and rapid fabrication of nano/microfluidic channels, (4) Modeling, analysis and characterization of the electronic properties of biomarker-aptamer interactions measured between the nano-electrodes, and, (5) Real-time low-power noise-free read-out circuit with sequential addressing, actuation, measurement & data analysis of the recognition sites.INTELLECTUAL MERITS: This proposal will transform and create a new area ?proteonics?, building up on the advances in ?proteomics? and ?molecular electronics?. The activities leverage from the molecular scale devices and the in vitro aptamer-protein interactions, and are extendible to a host of other applications. The ideas will overcome bottlenecks of expensive and serial fabrication in molecular electronics and provide alternate to the labor-intensive, poorly-sensitive and lengthy protocols of proteomics. The novel polymer nano/microfluidics will provide proper conditions to retain protein expression and functionality. On-chip circuit will lead the way to prototype point-of-care proteonic bio-chips. The nano-electrodes will provide a 3-D interaction volume for aptamer-protein binding, resulting in higher sensitivity and signal-to-noise ratio than those for planar morphologies. The approach will also overcome sensitivity limitations by removing the effects of device doping, geometry, dimensions, and fluidic environments. The proposed strategies will innovatively transform and revolutionize a number of disciplines: (1) Rapid nano-manufacturing for bio-sensing, (2) Multiplexed detection of disease markers using various aptamers, (3) Ultrasensitive on-chip electrical detection of biomarkers and analysis for early disease detection, (4) Mask-less production of novel nano/microfluidics. BROADER IMPACT: The proposal has direct applications in other biosensor domains, e.g. gene expression analysis, virus/pathogen detection and whole blood analysis. The variations of the propose technology can transform biomolecular sensing with better disease intervention strategies, improved statistical confidence and real-time detection. The PI has engaged women graduate students and minority undergraduate/high school students in his research lab. Innovative educational endeavors will be pursued with this proposal: (1) Development of a graduate course on nano-bio devices, (2) Seminars/Demos/Lab-tours focused on research involvement and retention of undergraduates, (3) One-week summer camp for high school students (primarily African-American and Hispanic) from Arlington school district, integrating MEMS/Nano research and biology concepts, (4) Interactive website/blog for the projection/exposure/discussion of the state of the art in research, (5) Saturday morning live-chat sessions to follow-up/engage K-12 students and teachers, (6) Technology transfer studies to nurture entrepreneurship in students interested in real-world problems, (7) Development of international research collaborations for exchange of students from and to USA. The results of the proposed ideas will be disseminated through peer-reviewed articles, conferences and public media.