Single-Molecule Studies of Proteins at Polymer based Chromatographic Interfaces
Moringo, Nicholas A
Landes, Christy F
Doctor of Philosophy
The worldwide pharmaceutical landscape has witnessed a large influx of biological-based therapeutics, termed ‘biologics’, for the successful treatment of diseases and illnesses. The downstream separation and purification of promising biologics via chromatography dominates the total production cost leading to market entry. A mechanistic examination of protein interactions at the interface of stationary phase materials can improve and enable predictive chromatographic separation optimization. Single-molecule imaging techniques, inspired by the advances of super-resolution microscopy, can capture the highly dynamic interactions of proteins at stationary phase materials. It is observed that nanoscale protein dynamics can explain experimentally observed increases in separation efficiencies. We uncover how the suppression of anomalous surface diffusion, which leads to improved separations, can be tuned with stationary phase surface chemistries, polymer packing, and ionic salt conditions. Overall, we have shown that single-molecule imaging can relate protein dynamics at the nanoscale to improved protein separations at the interface of synthetic polymer materials.
single-molecule microscopy; chromatography