Structural characteristics of low-molecular-mass displacers for cation-exchange chromatography

The relative efficacy of a variety of low-molecular-mass displacers was examined using a displacer ranking plot. This method enables an evaluation of the dynamic affinity of a variety of displacers over a range of operating conditions. Several homologous series of molecules were evaluated to provide insight into the effects of various structural features on displacer efficacy. The results indicate that linear flexible geometries may have advantages over branched or cyclic structures. Data also indicate that the spreading out of charges may increase affinity. The incorporation of aromatic moieties in these displacers, particularly near the surface of the molecules, appears to result in a dramatic increase in displacer affinity. The ability of several high-affinity low-molecular-mass displacers a very strongly bound cationic protein is also examined. The results confirm the predictions of the theory and indicate that it is indeed possible to displace highly bound macromolecules with low-molecular-mass dispatchers. The work presented in this paper indicates that non-specific interactions can be exploited for producing high-affinity low-molecular-mass displacers.