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：自包含、高通量、超灵敏的多路生物分子传感技术[j]: Katsuo kurabayashi]: Ku Pei-Cheng， Edgar meyhofero自然界进化了数百万年的高效分子机器？应用于微型分析系统中，在生物传感、分子分选、流体泵送、微机械驱动和分子组装等生物纳米技术应用中发挥着重要作用。它们具有纳米尺寸的紧凑性，在流体环境中产生强大的运动，并且易于由含有三磷酸腺苷（ATP）的溶液提供燃料。这消除了对微/纳米流体驱动的外部能源的需要。此外，bmm有效地操纵单个生物分子和蛋白质，使基于运动蛋白的具有纳米级质量传递/浓度功能的生物传感系统的发展成为可能。这项由美国国家科学基金会颁发的生物传感/CBET项目奖支持密歇根大学Kurabayashi、Ku和Meyhofer教授开发的一种新的生物传感芯片技术，即生物分子马达（BMM）智能微阵列，它允许高通量、超灵敏（原子摩尔浓度）的生物传感用于芯片上的多路蛋白质结合分析。在微流控系统中结合基于BMM的质量传输/传感机制，BMM智能微阵列能够自主处理样品，包括通过马达蛋白驱动的微管对多个目标分析物进行特定的结合、分选、运输和浓缩。所提出的方法将生物分子马达、光子学和纳米流体结合在一个单一的生物传感器中，同时将大量的分子分析物输送和浓缩到特定的检测器上，以进行超灵敏的定量分析。通过开发高通量超灵敏的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。提出的结合生物纳米技术和基于led的固态照明技术进行超灵敏多路蛋白质传感的基础研究，最终可以扩展到通过非常简单和强大的电池操作的手持模块设置来实现疾病的早期检测。这可能为在资源有限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pi吸收。纳米生物力学，微机电系统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与多个领域的研究人员合作，获得MEMS技术、微纳米制造、生物物理学、生物化学和光子学方面的综合知识和技能。的学生吗?通过在国家/国际MEMS和纳米技术会议上的演讲，他们的沟通和网络技能将得到提高，研究将被纳入pi ？MEMS和纳米制造的跨学科研究生课程。由美国国家科学基金会支持的国家纳米技术基础设施网络（NNIN）本科生研究经验项目（REU）将积极招募来自代表性不足群体的暑期实习生。这项研究具有革命性的潜力，因为它(1)首次展示了将基于bmp的纳米级质量传输和生物传感用于多路生物传感的技术；(2)利用先进的科学知识，在人造工程结构中控制bmm，并在低浓度样品中获得高信噪比的弱生物荧光信号，为实现稳健、经济、简单的临床诊断提供了一种新的方法。

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.