Micro and Nanofluidic Methods for Studying Supramolecular Assembly

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This project focuses on the development of micro and nanofluidic methods for the study of supramolecular protein assembly. We will aim to identify within a complex mixture proteins and protein complexes by trapping them, using electrophoretic, electrostatic and hydrodynamic forces, at nanojunctions on microfluidic devices fabricated using soft lithographic techniques. This approach will allow the selective enriching of species of interest and will thus allow us to reach very high sensitivities even with dilute analyte solutions, allowing minority populations of misfolded proteins and incorrectly assembled protein complexes to be identified. As part of the project we will develop approaches for effective integration of nano and microfluidic elements on chip. This objective will be achieved through the use of two-photon lithography to overcome the diffraction limit through non-linear optics and to define nanoscale features in the same photoresist used for conventional microfabrication. The lithographically patterned substrate will then be used to imprint polymers in a soft lithography process to yield a scalable avenue to fabricate devices integrating nano and micro scale features. We will detect proteins through deep UV fluorescence, which will allow us to work with fully unlabelled species. These results will open up the path towards quantitative charaterisation of protein self-assembly at high resolution in their native environment.