IDBR: Development of Ultrasensitive, Superfast, and Microliter-Volume Differential Scanning Nanocalorimeter for Direct Characterization of Biomolecular Interactions

摘要：开发用于直接表征生物分子相互作用的超灵敏、超快速、微容积差示扫描纳米热仪所提出的差示扫描纳米热仪将大大减少样品体积，缩短分子现象的测量时间，为全面的高含量热力学研究、膜蛋白研究和药物发现早期提供有力的工具。这项多学科研究将为年轻科学家和工程师，特别是妇女和代表性不足的少数民族提供极好的机会。与不同应用领域的国家实验室和行业的互动将加速开发知识的实施。所有的生物现象都依赖于分子间的相互作用，如配体与蛋白质的结合，或分子内的相互作用，如蛋白质折叠。作为一种无标记和无固定的方法，现代量热仪是直接表征分子相互作用热力学特征的金标准，包括吉布斯自由能、焓、熵、比热和化学计量学，为研究生物分子机制和药物设计提供了有价值的信息，这些信息不能单独从结构或计算方法中获得。然而，目前最先进的量热计需要大体积、高浓度的蛋白质，并且需要很长的测量时间，这限制了它们在生物学研究中的应用。该奖项的目标是开发一种创新的基于mems的差分扫描纳米热计和阵列，以将生物大分子的消耗从毫升减少到微升，并将测量时间从几小时减少到几分钟，从而实现对生物分子相互作用的直接，精确和快速检测。为了实现这一目标，我们计划了四项具体任务，包括1)设计和制造超灵敏低噪声传感器，2)最小化纳米热计的寄生热损失，3)设计和集成低噪声电子和反馈控制器，以及4)测试并将该技术应用于膜蛋白研究和药物筛选。

Award Abstract: Development of Ultrasensitive, Superfast, and Microliter-Volume Differential Scanning Nanocalorimeter for Direct Characterization of Biomolecular InteractionsThe proposed differential scanning nanocalorimeter will significantly reduce the sample volume and shorten the measurement time of molecular phenomena, and provide a powerful tool for comprehensive high-content thermodynamics studies, in the study of membrane proteins and in the early stage of drug discovery. This multidisciplinary research will provide excellent opportunities to young scientists and engineers, especially women and underrepresented minorities. Interaction with national laboratories and industry at different ends of the application spectrum will enable accelerated implementation of the developed knowledge.All biological phenomena depend on molecular interactions, which is either intermolecular, as with ligand binding to a protein, or intramolecular, as with protein folding. As a label-free and immobilization-free method, modern calorimetry instrumentation is the gold standard for directly characterizing the thermodynamic profiles of molecular interactions, including Gibbs free energy, enthalpy, entropy, specific heat, and stoichiometry, providing valuable information for investigating biolomolecular mechanisms and drug design, information that cannot be obtained from structural or computational methods alone. However, the current state-of-the-art calorimeters require large-volume, high-concentration proteins and need very long measurement times, which limits their utility for biological studies. The objective of this award is to develop an innovative MEMS-based differential scanning nanocalorimeter and array to reduce the consumption of biological macromolecules from milliliters to micro liters, and to decrease the measurement time from hours to minutes, and thus enable direct, precise, and rapid detection of biomolecular interactions. To achieve this goal, four specific tasks are planned, including 1) designing and fabricating ultrasensitive low-noise sensors, 2) minimizing the parasitic heat loss of the nanocalorimeter, 3) designing and integrating low-noise electronics and feedback controller, and 4) testing and applying this technology to membrane protein study and pharmaceutical screening.