Nanofluidics for label-free detection of exosomes and protein aggregates in neurodegenerative disease research

Microfluidics have become a powerful tool in biotechnology and life sciences, whereas the nanofluidic regime remains widely unused in industry and the clinical environment. Especially protein misfolding diseases such as Alzheimer’s, Parkinson’s and Huntington’s disease, recently experience a growing demand for single-molecule detection capabilities, as the assembly process of a single corrupted protein is correlated with its aggregation propensity and spread of the disease. Conventional microfluidic methods to study relevant biomarkers and aggregates involve fluorescent labelling, which alters the samples' properties and puts additional constraints on experimental design. We therefore seek for methods to study macromolecules (in particular: exosomes, oligomers) in a label-free manner in solution without chemically altering their properties.To overcome this limitation, the project combines innovative nanofluidic technology with cutting-edge label-free microscopy techniques. First, hybrid 2-photon lithography is used for the scalable cost-effective nanofabrication of nanofluidic polymer chips. Secondly, these chips are then employed for the detection and sizing of exosomes (<100 nm) and alpha-synuclein protein aggregates (<12 nm) in solution using label-free imaging methods (e.g. QPM and DUVM).The approach presented here, allows protein misfolding researchers to study oligomerization more efficiently by running orders of magnitude more experiments with same already limited material, and greatly increases availability of nanofluidic chips for protein metrology in existing biological laboratories. New expertise, excellence, management skills and scientific training acquired during the action, prepare the researcher for a role as independent dementia research group leader in Europe, using state-of-the-art nanotechnology to establish nanofluidic single-molecule detection of protein aggregates and macromolecules as new golden golden standard in life sciences research.