Identification of Conformation Differences between Protein Isoforms for Developing Resolute Separation Methods

The purpose is to develop approaches to separate protein isoforms through the identification of conformation differences of one isoform relative to the other. Development of approaches to separate protein isoforms is propelled by the central role that these macromolecules have been found to have in the pathological states of disease and mutations. Additional Motivational Factor: Current Methods of protein separation require unrealistically large amounts of highly concentrated protein and a significant amount of time; microfluidic approaches can sidestep such requirements. Current methodologies for separating protein isoforms attempt to separate protein molecules while in their native or fully denatured state. Knowing that proteins adopt different conformation pathways as a function of the solvent environment they are equilibrated in, it is hypothesized that a particular buffer in which distinct conformations differences arise between two isoforms will provide for more resolute separation. Through the identification of buffer conditions in which protein isoforms conformations are significantly different relative to one another, separation of close mobility molecules can be made extremely resolute since such conditions will provide an environment in which differences in electrophoretic mobility and/or adsorption kinetics become more discernable and exploitable.