Enthalpy arrays for thermodynamic characterization

用于热力学表征的焓阵列

基本信息

批准号：
7628512
负责人：
Richard H. Bruce
金额：
$ 36.63万
依托单位：
PALO ALTO RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-06-01 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7628512
关键词：
Address Affinity Area Basic Science Binding Biochemical Biochemistry Biological Assay Biology CD209 gene Calorimetry Carbohydrates Cells Chemicals Collaborations Commit Consumption DNA Data Detection Development Discipline Disclosure Engineering Ensure Entropy Free Energy Funding Generic Drugs Goals HIV HIV Infections HIV Protease HIV Protease Inhibitors Immobilization Immune system Label Letters Life Cycle Stages Ligand Binding Ligands Measurement Measures Mechanics Methods Microfabrication Modification Molecular Noise Nucleic Acids Physics Production Proteins Proteomics RNA Reaction Reagent Research Research Personnel Sampling Screening procedure Senior Scientist Surface System Techniques Technology Thermodynamics Time Titrations Work assay development base chemical reaction cost design detector drug discovery enthalpy evaporation experience improved inhibitor/antagonist instrument interest intermolecular interaction meetings microcalorimetry multidisciplinary neutralizing antibody programs receptor sensor small molecule stoichiometry technology development tool

项目摘要

DESCRIPTION (provided by applicant): A generic method to characterize molecular interactions with no assay development would substantially benefit both functional proteomics research and drug discovery; however, none of the available approaches offers the efficient detection and low reagent consumption needed for the large number of measurements. This proposal addresses developing a sensitive enthalpic assay tool to detect bimolecular interactions involving proteins, ligands, RNA, DNA and other molecules of interest without requiring any special assay development. In particular, the assay does not require development of molecule-specific inhibitors, probes, detectors, attachment of labels, immobilization on surfaces, or chemical modifications of the molecules. The assay is based on direct detection of the enthalpy released during interaction of the species of interest. The detector, a nanocalorimeter, will enable measurement of interactions for molecular concentrations in the low micro-molar range using sample volumes of 200 nl. The technical goal of this work is to develop a detector that can detect enthalpies in the nanocalorie range with an effective measurement time of six seconds. This represents nearly three orders of magnitude reduction in the required sample and measurement time compared with conventional microcalorimetry. We propose to develop an array of 96 detectors and use it to study key aspects of HIV infection and life cycle. In addition to detecting interactions the enthalpy array can be used to perform titrations to determine binding constants. The effort integrates several engineering disciplines with applied physics and biochemistry. We have developed the initial proof of concept through a multidisciplinary effort bridging microfabrication with several engineering disciplines (mechanical, electrical chemical and system) as well as applied physics. We are collaborating with several senior biochemists to ensure that the system meets their scientific needs.

描述（由申请人提供）：一种表征分子相互作用而无需测定开发的通用方法将显着有益于功能蛋白质组学研究和药物发现；但是，所有可用方法都没有提供大量测量所需的有效检测和低试剂消耗。该建议旨在开发一种敏感的焓测定工具，以检测涉及蛋白质，配体，RNA，DNA和其他感兴趣的分子的双重分子相互作用，而无需任何特殊的测定开发。特别是，该测定不需要开发分子特异性抑制剂，探针，检测器，标记的附着，固定在表面上或分子的化学修饰。该测定基于对感兴趣物种相互作用期间释放的焓的直接检测。检测器（一种纳米氧计）将使用200 nL的样品体积来测量低微摩尔范围内分子浓度的相互作用。这项工作的技术目标是开发一个检测器，该检测器可以通过六秒钟的有效测量时间在纳米级范围内检测焓。与常规微钙化法相比，这代表了所需样品的近三个数量级减少和测量时间。我们建议开发96个检测器的阵列，并使用它来研究HIV感染和生命周期的关键方面。除了检测相互作用外，焓阵列可用于执行滴定以确定结合常数。这项工作将几个工程学科与应用物理和生物化学相结合。我们通过多个工程学科（机械，电气化学和系统）以及应用物理学的多学科努力以及跨越的概念证明了概念证明。我们正在与几位高级生物化学家合作，以确保该系统满足他们的科学需求。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Higher throughput calorimetry: opportunities, approaches and challenges.

DOI：
10.1016/j.sbi.2010.09.001
发表时间：
2010-10
期刊：
CURRENT OPINION IN STRUCTURAL BIOLOGY
影响因子：
6.8
作者：
Torres, Francisco E.;Recht, Michael I.;Coyle, Joseph E.;Bruce, Richard H.;Williams, Glyn
通讯作者：
Williams, Glyn

Rapid mixing of sub-microlitre drops by magnetic micro-stirring.