A high-throughput multi-well dynamic light scattering instrument

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 520417342

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.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Ein Hochdurchsatz-Multiwell-Instrument für dynamische Lichtstreuung
• Instrumentation Group: 5360 Meßgeräte für gestreutes und reflektiertes Licht, optische Oberflächen-Prüfgeräte
• Applicant Institution: Heinrich-Heine-Universität Düsseldorf
• Leader: Professor Dieter Willbold, Ph.D.