A high-throughput multi-well dynamic light scattering instrument

一种高通量多孔动态光散射仪

• 批准号: 520417342
• 金额: --
• 依托单位: Heinrich-Heine-Universität Düsseldorf
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2023
• 资助国家: 德国
• 起止时间: 2022-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/520417342?language=en
• 关键词: throughput; multi; well; dynamic; light

We request a low-volume, multi-well dynamic light scattering (DLS) device equipped with an incubator plate hotel. The instrument differs from our current model and from other offerings on the market by its ability to work with extremely small sample volumes (< 0.1 µL), and the availability of a built-in microscope for imaging both in the UV-vis range. Compared to our current single-cuvette instrument, the plate-based concept of the proposed device is expected to speed up our experimentation by two orders of magnitude. In the interval between measurements, plates are stored in an attached temperature controlled hotel unit that is completely automated and can house multiple plates. This proposal is intended to support our research activities focusing on protein aggregation and its pathophysiological significance in neurological diseases. Essential objectives in these projects include the characterization of self- and hetero-association of proteins, their misfolding pathways and resulting aggregation states. In these studies, physico-chemical parameters such as temperature, pH, ionic strength, and other solvent properties represent important variables to be investigated. Protein aggregation and fibrillization will lead to an increase of the effective hydrodynamic radius of particles, with a concomitant increase in the scattered intensity and a slow-down in its fluctuation. Using kinetic models, we are then able to describe the aggregation mechanism of the protein of interest and its potential relevance to the pathophysiology of the respective disease. In another important branch of our research, we apply the knowledge acquired on protein aggregation in human diseases for development of therapeutics. Many of our candidate therapeutic agents are L- or D-enantiomeric peptide leads that were identified using phage display (PD) or mirror image PD technology. Here, DLS represents an efficient way of characterizing large sets of customised peptides under various different conditions, without the need for chemical labelling, e.g. with fluorescent dyes, which may significantly alter the properties of the compounds under investigation. It is important to note that aggregating systems are extremely sensitive to timing and environmental conditions, so ensuring reproducibility of results is particularly demanding. The capability of handling 96 samples in parallel on a single plate, combined with the precise scheduling of measurements and temperature control during incubation, as offered by the proposed device, will dramatically enhance the validity of the data acquired, and to allow for studies, which are not possibly with the present available device. Another important consideration relates to reproducibility and replication of experiments. This is calling for reduction of human intervention as much as achievable. An automated high-throughput platform as the one requested is considered the appropriate solution to cope with these and many other challenges.

我们要求一个低容量，多孔动态光散射（DLS）设备配备了一个孵化器板酒店。该仪器与我们当前的型号和市场上其他产品的不同之处在于，它能够处理极小的样品体积（< 0.1 µL），并且可以使用内置显微镜在紫外-可见光范围内进行成像。与我们目前的单试管仪器相比，所提出的设备的基于板的概念预计将加快我们的实验两个数量级。在测量间隔期间，将平板储存在一个完全自动化的温度控制的附属酒店单元中，该单元可以容纳多个平板。该提案旨在支持我们的研究活动，重点是蛋白质聚集及其在神经系统疾病中的病理生理学意义。这些项目的基本目标包括蛋白质的自我和异源缔合的表征，它们的错误折叠途径和由此产生的聚集状态。在这些研究中，物理化学参数，如温度，pH值，离子强度和其他溶剂性质代表了要研究的重要变量。蛋白质的聚集和纤维化将导致颗粒的有效流体动力学半径的增加，伴随着散射强度的增加和其波动的减缓。使用动力学模型，我们能够描述感兴趣的蛋白质的聚集机制及其与相应疾病的病理生理学的潜在相关性。在我们研究的另一个重要分支中，我们将所获得的关于人类疾病中蛋白质聚集的知识应用于治疗方法的开发。我们的许多候选治疗剂都是L-或D-对映体肽先导物，这些先导物是使用噬菌体展示（PD）或镜像PD技术鉴定的。在这里，DLS代表了一种在各种不同条件下表征大量定制肽的有效方法，而不需要化学标记，例如用荧光染料，这可能会显着改变所研究化合物的性质。重要的是要注意，聚集系统对时间和环境条件极其敏感，因此确保结果的可重复性特别重要。在单个平板上平行处理96个样本的能力，结合孵育期间测量和温度控制的精确时间表，如所提出的设备所提供的，将显著提高所获取数据的有效性，并允许进行研究，这是目前可用的设备所不可能的。另一个重要的考虑因素涉及实验的再现性和重复性。这就要求尽可能减少人为干预。所要求的自动化高通量平台被认为是科普这些挑战和许多其他挑战的适当解决方案。