AFM Arrays for Single Molecular Mechanics

用于单分子力学的 AFM 阵列

• 批准号: 7023534
• 负责人: F. Levent Degertekin
• 金额: $ 29.38万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7023534
• 关键词: atomic force microscopy; bioengineering /biomedical engineering; bioimaging /biomedical imaging; microarray technology; nanotechnology

The immune system includes a wide variety of cells that communicate with each other via adhesion and signaling molecules. Many of these molecules behave as 'nanomachines' that connect, sense, move, actuate, to name a few of their functions. To understand the inner workings of such nanomachines requires the characterization of kinetic properties that govern their interactions. We have used atomic force microscopy (AFM) to directly measure mechanistically the behavior of several immune system receptors at the level of single pair of interacting molecules, which extends and complement the ensemble assays commonly used in biochemistry and biophysics. These experiments are laborious and time-consuming due to the requirement for a large ensemble of stochastic data. To address the throughput bottleneck, we will develop arrays of AFM with sufficient sensitivity, bandwidth, and low noise that are suitable for single molecule experiments. This will be done by integrating several existing technologies - cantilever array, photodetector array, and functionalized tip arrays on substrates with a new technology - acoustic radiation pressure (ARP) actuator array. The ARP-driven AFM arrays -protein chips - will enable a large number of parallel single molecular experiments at the same time instead of using a single AFM to conduct them sequentially over a long period of time. The ARP actuation technology will be tested in a single AFM and subsequently in the form of 1D and 2D AFM arrays for their ability to perform single molecule experiments. Using these AFM arrays, we will characterize the force regulation of kinetics of T cell receptor-ligand interactions. Better understanding of the properties and functions of these molecules may lead to the development of diagnostic and therapeutic strategies for immune system dysfunctions. The proposed research may also have significant commercial value because of the potential commercialization of the new AFM array technology. Moreover, protein chips such as this may provide means for the transduction of molecular recognition signals into mechanical, optical, and electrical signals through a natural interface to link individual biomolecular functions to microelectromechanical systems, which may have far-reaching implications to nanobiotechnology and biological/chemical warfare identification for national defense.

免疫系统包括各种通过粘附和信号分子相互通信的细胞。这些分子中的许多行为都是“纳米机器”，它们连接，感觉，移动，动摇，列举了一些功能。要了解这种纳米机器的内部起作用，需要对控制其相互作用的动力学特性的表征。我们已经使用原子力显微镜（AFM）在单对相互作用分子水平上直接测量了几种免疫系统受体的行为，该分子扩展并补充了在生物化学和生物物理学中常用的合奏测定法。由于需要大量随机数据集合，这些实验是费力的，而且耗时。为了解决吞吐量的瓶颈，我们将开发出适合单分子实验的足够灵敏度，带宽和低噪声的AFM阵列。这将通过集成几种现有技术 - 悬臂阵列，光电检测器阵列和官能化的尖端阵列， 新技术 - 声辐射压力（ARP）执行器阵列。 ARP驱动的AFM阵列 - 蛋白质芯片 - 将同时实现大量平行的单分子实验，而不是使用单个AFM在长时间内依次进行它们。 ARP驱动技术将以单个AFM进行测试，然后以1D和2D AFM阵列的形式进行测试，以便他们执行单分子实验的能力。使用这些AFM阵列，我们将表征T细胞受体配体相互作用动力学的力调节。更好地了解这些分子的特性和功能可能会导致免疫系统功能障碍的诊断和治疗策略的发展。拟议的研究也可能具有巨大的商业价值，因为新的AFM阵列技术的潜在商业化。此外，诸如此类的蛋白质芯片可以通过自然界面将分子识别信号转导到机械，光学和电信号中的蛋白质芯片，以将个体生物分子功能与微电机械系统联系起来，这可能具有极遥远的影响与纳米生物学和生物学/生物学//生物学//生物学/生物学//生物学/的影响。国防的化学战标识。