Previous studies have demonstrated that chemically selective displacement chromatography can be employed to add additional dimensions of selectivity to preparative ion exchange separations. Robotic high throughput screening was employed in concert with several analytical techniques (STD-NMR, SPR and MD Simulations) to identify and evaluate the behavior of chemically selective displacers for protein purification. These results show that selective displacement is achieved by incorporating a binding affinity for the targeted molecule into the design of the chemically selective displacer while also retaining the displacer's affinity for the resin. From these results fluorescent displacers were rationally designed and tested in order to facilitate online detection. Finally, a novel approach was developed for applying selective displacement for the removal of key industrial impurities from product streams. This work demonstrates the ability of chemically selective displacement chromatography to provide high resolution separations while simultaneously removing a targeted molecule, thus eliminating further downstream processing steps.