Collection, focusing, and metering of biomolecules using addressable microelectrode arrays for portable low-power bioanalysis

使用可寻址微电极阵列收集、聚焦和计量生物分子，进行便携式低功耗生物分析

• 批准号: 0554108
• 负责人: Victor Ugaz
• 金额: $ 25万
• 依托单位: Texas A&M Engineering Experiment Station
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0554108&HistoricalAwards=false
• 关键词: Collection; focusing; metering; biomolecules; using

Abstract: Advances in microfluidic technology have enabled increasingly sophisticated biosensing and bioassay operations to be conducted at the microscale, many of these applications employ such small amounts of DNA that it must first be pre-concentrated to a detectable level. Often the sample contains multiple components or is unknown, making it difficult to employ techniques like polymerase chain reaction (PCR) amplification to increase the concentration level. Efficient strategies for precisely handling minute quantities of biomolecules in microchannel geometries are critically needed, however it has proven challenging to achieve simultaneous concentration, focusing, and metering with current-generation technology. This research program will address these issues by exploring advanced microfluidic systems incorporating arrays of on-chip electrodes to digitally collect and meter DNA and other charged biomolecules in precise and controllable increments. This technique transports charged biomolecules between active electrodes upon application of a small potential (~1 V), and is capable of achieving orders of magnitude concentration increases within a small device footprint suitable for portable operation (a single AA battery may deliver sufficient power to perform multiple concentration and metering operations). The collected samples are highly focused, with sample zone size and shape defined solely by electrode geometry. The results of these fundamental studies will also be applied to explore entirely new first-of-their-kind applications for this technology including (i) coupling the capture process with a superimposed hydrodynamic flow to concentrate and re-suspend a DNA sample in a new buffer environment suitable for subsequent reactions or analysis, (ii) harnessing the electrode capture process to collect DNA in free solution followed by hydrodynamic injection of a gel sieving matrix directly over the collected DNA to perform subsequent electrophoretic separation, and (iii) capture and focusing of protein samples. These novel applications will address the compelling economic incentives for developing advanced DNA-based assay technologies that will be key components in next-generation genomic analysis systems. In addition to the commercial impact associated with making genomic analysis technology more affordable, this technology will greatly enhance education in emerging multidisciplinary areas of the Chemical Engineering discipline through integration with the PIs efforts to develop new educational experiences for undergraduates at the interface between the physical, chemical, and life sciences. The PI will also introduce a regular series of debates involving undergraduates, graduate students, and faculty to stimulate interest in the societal impact of bio- and nanotechnology issues while improving communication skills and providing a common experience that can be shared by the entire department. Students involved in this project will gain hands-on experience in areas at the frontiers of the Chemical Engineering discipline, and society will benefit through the development of new genomic analysis technology for improved health care.

摘要：微流控技术的进步使得越来越复杂的生物传感和生物测定操作能够在微尺度上进行，其中许多应用使用如此少量的DNA，以至于必须首先将其预浓缩到可检测的水平。通常，样品含有多种成分或未知，使得难以采用聚合酶链反应（PCR）扩增等技术来提高浓度水平。在微通道几何形状中精确处理微量生物分子的有效策略是迫切需要的，然而，事实证明，用当前的技术实现同时浓缩、聚焦和计量具有挑战性。该研究计划将通过探索先进的微流体系统来解决这些问题，该系统包含芯片上电极阵列，以精确和可控的增量数字化收集和计量DNA和其他带电生物分子。该技术在施加小电位（~ 1V）时在活性电极之间输送带电生物分子，并且能够在适合于便携式操作的小装置占地面积内实现数量级的浓度增加（单个AA电池可以输送足够的电力以执行多个浓度和计量操作）。所收集的样品是高度集中的，样品区的大小和形状仅由电极几何形状定义。这些基础研究的结果也将被应用于探索该技术的全新的首次应用，包括（i）将捕获过程与叠加的流体动力学流耦合，以将DNA样品浓缩并重新悬浮在适合于后续反应或分析的新缓冲液环境中，（ii）利用电极捕获过程收集游离溶液中的DNA，然后直接在收集的DNA上流体动力学注射凝胶筛分基质以进行随后的电泳分离，和（iii）蛋白质样品的捕获和聚焦。这些新的应用将解决开发先进的基于DNA的分析技术的令人信服的经济激励，这将是下一代基因组分析系统的关键组成部分。除了与使基因组分析技术更实惠相关的商业影响外，该技术还将通过与PI的整合，为本科生在物理，化学和生命科学之间的接口开发新的教育经验，大大加强化学工程学科新兴多学科领域的教育。PI还将引入一系列涉及本科生，研究生和教师的定期辩论，以激发对生物和纳米技术问题的社会影响的兴趣，同时提高沟通技巧，并提供可以由整个部门分享的共同经验。参与该项目的学生将获得化学工程学科前沿领域的实践经验，社会将通过开发新的基因组分析技术来改善医疗保健而受益。