The Development of a Chip-Scale Nano-Calorimeter

芯片级纳米量热仪的研制

• 批准号: 8118437
• 负责人: DALE NORMAN LARSON
• 金额: $ 25.3万
• 依托单位: CHARLES STARK DRAPER LABORATORY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8118437
• 关键词: Address; Affinity; Binding; Binding Sites; Biochemistry; Biology; Biophysics; Boston; Buffers; Calorimetry; Cells; Clinic; Data; Deposition; Detection; Development; Device Designs; Devices; Dimethyl Sulfoxide; Doctor of Philosophy; Engineering; Ensure; Entropy; Equilibrium; Equipment; Evaluation Studies; Event; Feedback; Free Energy; Gefitinib; Generations; Gleevec; Goals; Gold; Heating; Kinetics; Knowledge; Laboratories; Lateral; Ligands; Light; Liquid substance; Literature; Malignant Neoplasms; Measurement; Measures; Methods; Microfluidics; Modeling; Nature; Optics; Performance; Pharmacologic Substance; Pharmacology; Process; Productivity; Proteins; Publishing; Reaction; Reference Standards; Reference Values; Relative (related person); Research; Research Personnel; Role; Sampling; Screening procedure; Shapes; Signal Transduction; Solid; Surface; Surface Plasmon Resonance; System; Technology; Temperature; Testing; Thermodynamics; Thick; Time; Titrations; Travel; Universities; Work; analytical method; analytical tool; assay development; base; computerized data processing; design; drug development; enthalpy; experience; high throughput screening; innovation; instrumentation; multidisciplinary; nano; nanoscale; prototype; research and development; research study; sensor; stoichiometry; therapeutic target; tool; transmission process

DESCRIPTION (provided by applicant): The study of binding interactions is a central aspect of basic biology research and pharmaceutical R&D and there are numerous analytical methods available to study various aspects of these interactions. Each has its own strengths and weaknesses. Calorimetry is currently used, not as a screening tool, but as a tool to understand a specific reaction and is very important in the study of binding interactions. A calorimeter measures the energy released or absorbed by a reaction over a range of reactant concentrations to determine the relative contributions of enthalpically driven processes (related to the number and types of bonds) and entropically driven processes (related to the shapes of the binding site and the ligand). Unfortunately, the need for a large amount of protein (0.5 to 5mg) limits its usage. Additionally, there are some reactions where the amount of heat is too small for the current generation of calorimeters to measure. We are developing a chip scale calorimeter based on extraordinary optical transmission (EOT) through an array of nanometric apertures. Stark et al and Brolo et al have shown that these nanohole array devices can be used as affinity sensors where one of the binding partners is immobilized on the surface of the nanohole array device. With these nanohole array sensors the signal is temperature dependent due to the dielectric function of the buffer changing the plasmon excitation conditions. Holding the concentration constant in an approximately 100nm thick layer of dielectric directly above the nanohole array surface enables the use of EOT as a fast and sensitive temperature sensor to measure the heat of reaction (enthalpy, ¿H) from binding events. The inherent ability to multiplex many nanohole array sensor devices on a single chip enables the simultaneous measurement of controls to characterize confounding effects (e.g. buffer dilution, mixing, presence of DMSO in the buffer) and deconvolution of these effects to determine the true heat of reaction. This multiplexing also indicates the possibility of using this for high throughput screening as well as expanding on the current role of calorimetry. Early results indicate that a nanohole array calorimetry system has the potential to reduce the amount of protein required by 1000-fold and increase sensitivity by 100-fold. This will expand the use of calorimetry in pharmaceutical R&D. Our research plan consists of three specific aims to demonstrate proof-of-principle for this technology. Aims 1 and 2 explore the fundamental design options and tradeoffs involved in nanohole array device design and sample delivery. Aim 3 integrates these results into a calorimetry system and assesses the resulting measurement performance against quantitative milestones. In this application we propose to develop a new chip-scale nanocalorimeter that addresses the key limitations (compound usage, sensitivity, and analysis time) of current calorimetry technologies. The two primary performance goals for this project are to decrease compound usage by at least 1000-fold and to increase sensitivity by at least 100-fold while ensuring compatibility with existing liquid handling equipment.

描述（由适用提供）：结合相互作用的研究是基本生物学研究和药物研发的主要方面，并且有许多分析方法可用于研究这些相互作用的各个方面。每个人都有自己的优点和劣势。目前使用了量热法，而不是用作筛选工具，而是一种了解特定反应的工具，并且在研究结合相互作用中非常重要。量热计测量反应在一系列反应物浓度上释放或吸收的能量，以确定焓驱动的过程（与键的数量和类型有关）和熵驱动过程（与结合位点和配体的形状有关）的相对贡献。不幸的是，需要大量蛋白质（0.5至5mg）限制了其使用情况。此外，在某些反应中，热量太小，无法测量当前的Calolimeters。我们正在通过一系列纳米孔基于非凡的光学传输（EOT）开发芯片刻度量表。 Stark等人和Brolo等人表明，这些纳米荷尔阵列设备可以用作亲和力传感器，其中一个结合伙伴之一被固定在纳米霍尔阵列器件的表面上。使用这些纳米极阵列传感器，由于缓冲液的静脉功能改变了等离子兴奋条件，因此信号取决于温度。将浓度常数保持在大约100nm厚的旧层厚层中，直接在纳米尔阵列表面上方，可以将EOT用作快速和敏感的温度传感器，从而从结合事件中测量反应热（焓，€）。单个芯片上许多纳米荷尔阵列传感器设备的遗传性使控件的同时测量能够表征混杂效应（例如，缓冲稀释，混合，缓冲液中的DMSO存在）和这些效果的反向处理能够确定反应的真实热量。这种多路复用还表明将其用于高吞吐量筛选以及量热法的当前作用的可能性。早期结果表明，纳米荷尔阵列量热法具有将所需蛋白质量减少1000倍并提高敏感性100倍的潜力。这将扩大药品研发中量热法的使用。我们的研究计划包括三个特定的目的，旨在证明该技术的原理证明。 AIMS 1和2探索了纳米尔阵列设备设计和样品交付中涉及的基本设计选项和权衡。 AIM 3将这些结果集成到量热法系统中，并评估针对定量里程碑的结果测量性能。在此应用程序中，我们建议开发一种新的芯片尺度纳米级别，以解决当前量热法技术的关键局限性（复合用法，灵敏度和分析时间）。该项目的两个主要绩效目标是将复合用法降低至少1000倍，并将灵敏度提高至少100倍，同时确保与现有液体处理设备的兼容性。

项目成果

Dispersion of a Nanoliter Bolus in Microfluidic Co-Flow.

• DOI: 10.1088/0960-1317/24/3/034006
• 发表时间: 2014-03
• 期刊: Journal of micromechanics and microengineering : structures, devices, and systems
• 作者: Conway AJ;Saadi WM;Sinatra FL;Kowalski G;Larson D;Fiering J
• 通讯作者: Fiering J

A continuous flow microfluidic calorimeter: 3-D numerical modeling with aqueous reactants.

• DOI: 10.1016/j.tca.2014.09.024
• 发表时间: 2015-03-10
• 期刊: THERMOCHIMICA ACTA
• 影响因子: 3.5
• 作者: Sen, Mehmet A.;Kowalski, Gregory J.;Fiering, Jason;Larson, Dale
• 通讯作者: Larson, Dale