Probing interactions and phase separations of proteins, colloids and polymers with light scattering
The broad objective of my research is to investigate the physical characteristics and interactions of macromolecules and nanoparticles, and the corresponding effects on their phase separation behavior using static and dynamic light scattering (SLS & DLS). Light scattering provides a non-invasive...
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| Format: | Others |
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Scholar Commons
2009
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| Online Access: | http://scholarcommons.usf.edu/etd/2129 http://scholarcommons.usf.edu/cgi/viewcontent.cgi?article=3128&context=etd |
| Summary: | The broad objective of my research is to investigate the physical characteristics and interactions of macromolecules and nanoparticles, and the corresponding effects on their phase separation behavior using static and dynamic light scattering (SLS & DLS). Light scattering provides a non-invasive technique for monitoring the in-situ behavior of solutes in solution, including solute interactions, sizes, shapes, aggregation kinetics and even rheological properties of condensed phases. Initially, we investigated lysozyme solutions for the presence of preformed aggregates and clusters that can distort the kinetics of protein crystal nucleation studies in this important model system for protein crystallization. We found that both undersaturated and supersaturated lysozyme solutions contained population of large, pre-existing protein aggregate.
Separating these clusters and analyzing their composition with gel chromatography indicated that these clusters represented pre-formed lysozyme aggregates, and not extrinsic protein contamination. We investigated the effect of chaotropic versus kosmotropic ions (water structure breakers vs. structure makers) on the hydration layer and hydrodynamic interactions of hen egg white lysozyme. Surprisingly, neither chaotropic nor kosmotropic ions affected the protein hydration layer. Salt-effects on direct and hydrodynamic protein interactions were determined as function of the solutions ionic strength and temperature. Using both static and dynamic light scattering, we investigated the nucleation of gold nanoparticles forming from supersaturated gold sols. We observed that two well separated populations of nuclei formed essentially simultaneously, with sizes of 3nm vs. several tens of nanometer, respectively.
We explore the use of lysozyme as tracer particle for diffusion-base measurements of electrolyte solutions. We showed that the unusual stability of lysozyme and its enhanced colloidal stability enable viscosity measurement of salts solutions at high salt concentration, over a wide range of pH values and temperatures for the common tracer particle polystyrene flocculates. We applied dynamic light scattering to measure the viscoelastic responses of polystyrene probe particles embedded in solutions and gels of two different polymers: polyacrylamide (PAAm) and poly-N-isopropylacrylamide (poly-NiPAAm). |
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