Towards predictive protein separations: Imaging protein dynamics at nanoscale interfaces
Recent efforts by our group and others have shown the promise of applying single molecule methods to link mechanistic detail about protein adsorption to macroscale observables. When we study one molecule at a time, we eliminate ensemble averaging, thereby accessing underlying heterogeneity. However, we must develop new methods to increase information content in the resulting low density and low signal-to-noise data and to improve space and time resolution.
I will highlight recent advances in super-resolution microscopy for quantifying the physics and chemistry that occur between target proteins and stationary phase supports during chromatographic separations. My discussion will concentrate on the newfound ability of super-resolved single protein imaging to inform theoretical parameters via quantification of adsorption-desorption dynamics, protein unfolding, and nano-confined transport. Additionally, I will discuss using phase manipulation to encode temporal and 3D spatial information, and the opportunities and challenges associated with such imaging methods.