CAREER: Bio-Inspired Transport Assays and Single-Molecule Detection using Micro-and Nanopores

职业：使用微孔和纳米孔进行仿生运输测定和单分子检测

• 批准号: 0449088
• 负责人: Michael Mayer
• 金额: --
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0449088&HistoricalAwards=false
• 关键词: CAREER; Bio; Inspired; Transport; Assays

0449088MayerThe proposed research will focus on two related, bio-inspired projects. In one project, wells of volumes comparable to living cells (~10 12 liters) will be fabricated in silicon, glass, or polymer substrates. The open side of these microwells will be closed by a biomembrane. The Principal Investigator (PI) will then apply Nature's concept of using small, enclosed volumes to detect the in- or outflux of only a few molecules or ions. Like cells, the proposed setups will use multiple modes of amplification to detect the rapid rise in concentration after influx of less than 10,000 molecules through lipid bilayer membranes. These setups will be ideally suited to follow (i) passive transport of therapeutic drugs through lipid bilayers, (ii) activation of transmembrane receptors such as G protein-coupled receptors, and (iii) ion channel activity. Combining microfluidics with electrical and single-molecule fluorescence measurements might make it possible to (i) elucidate the time delay from binding of a fluorescent ligand to an ion channel to the opening of the pore in these proteins and (ii) investigate clinically relevant transporter proteins on a single molecule level.In the other project, The PI will use nanofabrication by femtosecond-pulsed lasers to take the bio-inspired approach one step further. The biological lipid membrane will be replaced with a robust, synthetic quartz membrane that supports a single nano-machined pore. In analogy to the gating characteristics and specificity of ion channels, the wall of the nanopore will be modified to attain specificity for antibodies, antigens, or viruses. Binding of those biomolecules might gate the current flux through the nanopore and result in more sensitive and diagnostically conclusive clinical assays than standard technology.

0449088梅尔拟议的研究将集中在两个相关的，生物启发的项目。 在一个项目中，威尔斯的体积相当于活细胞（~10 - 12升）将在硅，玻璃或聚合物基板制造。 这些微孔的开口侧将被生物膜封闭。 然后，主要研究者（PI）将应用大自然的概念，使用小的封闭体积来检测仅少数分子或离子的流入或流出。 像细胞一样，拟议的设置将使用多种扩增模式来检测通过脂质双层膜流入少于10，000个分子后浓度的快速上升。 这些设置将理想地适合于遵循（i）治疗药物通过脂质双层的被动转运，（ii）跨膜受体如G蛋白偶联受体的活化，和（iii）离子通道活性。 将微流体技术与电学和单分子荧光测量相结合，可以（i）阐明从荧光配体与离子通道结合到这些蛋白质中的孔打开的时间延迟，以及（ii）在单分子水平上研究临床相关的转运蛋白。PI将通过飞秒脉冲激光使用纳米纤维，将生物启发的方法更进一步。 生物脂质膜将被一个坚固的合成石英膜所取代，该膜支持一个纳米加工孔。 类似于离子通道的门控特性和特异性，纳米孔的壁将被修饰以获得对抗体、抗原或病毒的特异性。 这些生物分子的结合可能会控制通过纳米孔的电流，并导致比标准技术更灵敏和诊断结论性的临床测定。