The Marie Skłodowska Curie Action “A Single-Molecule Technology for Resolving Chaperone Action in Neurodegenerative Diseases” (project acronym: MicroSPARK) was devoted to advance our technological capabilities of studying misfolding processes in neurodegenerative diseases and elucidate mechanisms of chaperone action and drug action. This was achieved through the first-time combination and integration of single molecule and microfluidics technologies, and their use in dissecting the action mechanism of chaperones.
Neurodegenerative diseases, including Alzheimer’s and Parkinson’s disease, are increasingly prevalent disorders of our ageing society. These diseases arise from the formation of amyloidogenic protein aggregates. Molecular chaperones can counteract aggregate formation, but their molecular action mechanisms remain poorly understood. This Marie Skłodowska Curie Action addresses this challenge and implements the µSPARK technology to extend our understanding of the role of chaperones in the suppression of amyloid proliferation and in aggregate clearance, and introduces new means to sense amyloidogenic species for the development of therapeutics against neurodegenerative disease.
The core advancement of this project is the implementation of microfluidic sorting devices with single molecule detection to address important questions related to protein misfolding diseases with a specific focus on chaperones and diagnostics. Specifically, microfluidic diffusional sizing combined with confocal fluorescence spectroscopy was used to study the disaggregation mechanism of the Hsc70 heat shock protein machinery. Moreover, free-flow electrophoresis was used in combination with laser-induced spectroscopy to resolve heterogeneities of oligomeric species created during aggregation reactions. In addition, the technological capabilities of the platform were used to establish a new diagnostic technology termed digital immuno-sensing assay (DigitISA).
