A high-throughput multi-well dynamic light scattering instrument

我们需要一个小体积，多孔动态光散射（DLS）设备配备一个培养板酒店。该仪器与我们目前的型号和市场上的其他产品的不同之处在于，它能够处理极小的样品量（< 0.1µL），并且内置了一个显微镜，可在紫外-可见范围内进行成像。与我们目前的单比色皿仪器相比，所提出的装置的基于板的概念有望将我们的实验速度提高两个数量级。在测量之间的间隔，印版被储存在一个附加的温度控制的旅馆单元中，该单元是完全自动化的，可以容纳多个印版。本提案旨在支持我们专注于蛋白质聚集及其在神经系统疾病中的病理生理意义的研究活动。这些项目的基本目标包括蛋白质的自结合和异结合的表征，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是需要研究的重要变量。蛋白质聚集和成纤维化会导致颗粒有效水动力半径增大，同时散射强度增大，其波动速度减慢。利用动力学模型，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，我们将在人类疾病中获得的蛋白质聚集知识应用于治疗方法的开发。我们的许多候选治疗剂是L-或d -对映体肽引线，使用噬菌体展示（PD）或镜像PD技术鉴定。在这里，DLS代表了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，例如荧光染料，这可能会显著改变所研究化合物的性质。值得注意的是，聚合系统对时间和环境条件非常敏感，因此确保结果的可重复性是特别需要的。在单个平板上并行处理96个样品的能力，加上所提议的设备提供的精确测量计划和孵育期间的温度控制，将大大提高所获得数据的有效性，并允许进行目前可用设备不可能进行的研究。另一个重要的考虑因素与实验的可再现性和重复性有关。这就要求尽可能减少人为干预。一个自动化的高吞吐量平台被认为是应对这些挑战和许多其他挑战的合适解决方案。

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.