Biomolecular Motor Smart Microarrays: Self-Contained, High-Throughput, Ultrasensitive Multiplexed Biomolecular Sensing

摘要生物分子马达智能微阵列：自容式、高通量、超灵敏的多路生物分子传感PI：Katsuo Kurabayashi合作PI：Pei-Cheng Ku，埃德加Meyhofer近年来，生物分子马达（BMOs）？大自然进化了数百万年的高效分子机器已经被用于小型化分析系统中，并且在生物纳米技术应用中发挥重要作用，例如生物传感、分子分选、流体泵送、微机械动力和分子组装。它们是紧凑的，具有纳米尺寸，在流体环境中产生稳健的运动，并且容易由含有三磷酸腺苷（ATP）的溶液提供燃料。这消除了对用于微/纳流体致动的外部能量源的需要。此外，Bethesda有效地操纵单个生物分子和蛋白质，使得开发具有纳米级质量传输/浓缩功能的基于马达蛋白质的生物传感系统成为可能。生物传感/CBET计划的NSF奖项支持密歇根大学Kurabayashi，Ku和Meyhofer教授在开发新的生物传感芯片技术方面的研究，即生物分子马达（BMM）智能微阵列，该微阵列允许高通量，超灵敏（在阿托摩尔浓度）生物传感用于多路芯片上蛋白质结合测定。BMM智能微阵列在微流控系统中构建了基于BM的质量传输/传感机制，使自主样品处理成为可能，该自主样品处理涉及通过驱动蛋白马达蛋白驱动的微管特异性结合、分选、传输和浓缩多种靶分析物。该方法将生物分子马达、光子学和纳米流体学结合在一个生物传感器中，同时将大量（10个）分子分析物传输并浓缩到特定检测器中进行超灵敏定量，通过开发高通量超灵敏多路生物分子传感方法，将对分层医学和个性化医学等临床应用产生更广泛的影响。目标多路生物传感技术将允许监测疾病的早期微妙发作和生物威胁的早期预警。将生物纳米技术和基于LED的固态照明技术结合起来进行超灵敏多路复用蛋白质传感的基础研究最终可以扩展到使用非常简单和强大的电池供电手持模块设置来早期检测疾病。这可能为在有限资源的环境下开发用于即时护理应用的新商业产品打开大门。从这项研究中获得的基本知识将被吸收的PI？纳米生物力学、MEMS和光电器件技术的研究生课程。在这个项目中，参与的研究生和本科生将接受培训，以获得MEMS技术，微/纳米制造，生物物理学，生物化学和光子学的综合知识和技能，与多个领域的研究人员合作。学生？沟通和网络技能将通过他们在国家/国际MEMS和纳米技术会议上的演讲而增长，研究将被纳入PI？MEMS和纳米制造的跨学科研究生课程。 来自代表性不足群体的暑期实习生将通过NSF支持的国家纳米技术基础设施网络（NNIN）本科生研究经验计划（REU）积极招募到该项目中。这项研究代表了变革的潜力，因为它（1）提出了第一种技术，证明了使用基于BM的纳米级质量传输和生物传感的多路生物传感;以及（2）提供了一种新的方法来实现鲁棒的，成本有效的，简单的护理点临床诊断与先进的科学知识，控制人体内的细菌，工程结构，并在获得低浓度样品在高信噪比的微弱生物荧光信号。

AbstractBiomolecular Motor Smart Microarrays: Self-Contained, High-Throughput, Ultrasensitive Multiplexed Biomolecular SensingPI: Katsuo KurabayashiCo-PIs: Pei-Cheng Ku, Edgar MeyhoferIn recent years, biomolecular motors (BMMs) ? highly efficient molecular machines that nature has evolved for over millions of years ? have been employed in miniaturized analysis systems and play important roles in bionanotechnology applications, such as biosensing, molecular sorting, fluidic pumping, micromechanical powering, and molecular assembly. They are compact with a nanometer size, yield robust movement in a fluidic environment, and are readily fueled by adenosine triphosphate (ATP) containing solution. This eliminates the need for an external energy source for micro/nanofluidic actuation. In addition, BMMs efficiently manipulate individual biological molecules and proteins, making possible the development of a motor protein-based biosensing system with a nanoscale mass transport/concentration function. This NSF award by Biosensing/CBET program supports research by Professors Kurabayashi, Ku, and Meyhofer at the University of Michigan on the development of a new biosensing chip technology, namely the biomolecular motor (BMM) smart microarrays, which allows high-throughput, ultrasensitive (at attomolar concentrations) biosensing for multiplexed on-chip protein binding assays. Incorporating a BMM-based mass transport/sensing mechanism in a microfluidic system, the BMM smart microarrays enable autonomous sample handling that involves specific binding, sorting, transporting, and concentrating of multiple target analytes via kinesin motor protein-driven microtubules. The proposed method combines biomolecular motors, photonics and nanofluidics in a single biosensor to simultaneously transport and concentrate large numbers (10) of molecular analytes to specific detectors for ultra-sensitive quantification.The proposed effort will have broader impact on clinical applications such as stratified medicine and personalized medicine through developing a high-throughput ultra-sensitive multiplexed biomolecular sensing method. The aimed multiplexed biosensing technology will allow for monitoring of the early-stage subtle onset of diseases and early warning of biological threats. The proposed fundamental studies towards combining bionanotechnology and LED-based solid state lighting technology for ultrasensitive multiplexed protein sensing can ultimately be extended to enable the early detection of diseases with a very simple and robust battery-operated handheld module setting. This may open the door for the development of a new commercial product for point-of-care applications under an environment of limited resources. The fundamental knowledge gained from this research will be assimilated the PIs? graduate courses on nanobiomechanics, MEMS, and photoelectronic device technology. In this project, the involved graduate and undergraduate students will be trained to obtain integrated knowledge and skills in MEMS technology, micro/nano manufacturing, biophysics, biochemistry, and photonics, in collaboration with researchers across several fields. The students? communication and networking skills will grow through their presentations at national/international MEMS and Nanotechnology conferences, and research will be incorporated in the PIs? interdisciplinary graduate courses in MEMS and Nanomanufacturing. Summer interns from underrepresented groups will be actively recruited to this project through the National Nanotechnology Infrastructure Network (NNIN) Research Experience for Undergraduates Program (REU) supported by the NSF. This research represents transformative potential because it (1) presents the first technique that demonstrates the use of BMM-based nanoscale mass transport and biosensing for multiplexed biosensing; and (2) provides a new approach to realizing robust, cost-effective, simple point-of-care clinical diagnostics with advanced scientific knowledge on controlling BMMs within a man-made engineering structure and on obtaining weak biofluorescent signals at a high signal-to-noise ratio for low-concentration samples.