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

这项建议的目的是开发新的模式来分离和检测选择性蛋白质(生物标记物)，通过在具有多路纳米级电极的纳米/微流控通道/腔中利用适体-蛋白质相互作用和芯片上的数据处理。为了实现这些目标，将采用一致的制造和建模策略：(1)开发用于分离低丰度疾病生物标记物的功能化膜，(2)设计和开发具有可单独寻址的纳米电极的生物芯片，该生物芯片利用高通量纳米压印光刻制造并利用适体功能化用于生物标记物的多重检测，(3)开发新型和快速制造纳米/微流控通道，(4)对测量到的纳米电极之间的生物标记物-适体相互作用的电子性质进行建模、分析和表征，以及，(5)具有时序寻址、驱动、测量和放大功能的实时低功耗无噪声读出电路；识别位点的数据分析。互选优点：这项提议将改变和创建一个新的领域？蛋白质组学？，建立在蛋白质组学的进展？还有？分子电子学？这些活性利用了分子尺度装置和体外适配子-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中昂贵和系列化制造的瓶颈，并提供替代劳动密集型、低敏感性和冗长的蛋白质组学方案。新型聚合物纳米/微流控流体将为保持蛋白质的表达和功能提供合适的条件。片上电路将引领医疗保健点蛋白质生物芯片的原型。纳米电极将为适配子-蛋白质结合提供三维相互作用体积，从而比平面形态的电极具有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。拟议的战略将创新性地转变和革新一些学科：(1)用于生物传感的快速纳米制造，(2)使用各种适配子的疾病标志物的多重检测，(3)生物标志物的超灵敏芯片上电子检测和用于早期疾病检测的分析，(4)新型纳米/微流体的无掩膜生产。更广泛的影响：该提案在其他生物传感器领域也有直接应用，例如基因表达分析、病毒/病原体检测和全血分析。拟议技术的变化可以用更好的疾病干预策略、更好的统计置信度和实时检测来改变生物分子传感。PI在他的研究实验室聘请了女性研究生和少数族裔本科生/高中生。这项提议将推动创新的教育努力：(1)开发关于纳米生物设备的研究生课程，(2)以本科生参与研究和留住学生为重点的研讨会/演示/实验室之旅，(3)为期一周的夏令营，面向来自阿灵顿学区的高中生(主要是非裔美国人和西班牙裔)，将MEMS/Nano研究和生物学概念结合起来，(4)互动网站/博客，用于投射/暴露/讨论最新的研究成果，(5)星期六上午的实时聊天会议，以跟踪/接触K-12学生和教师，(6)技术转移研究，以培养对现实世界问题感兴趣的学生的创业精神；(7)发展国际研究合作，以交换来往美国的学生。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.