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Displacement Chromatography of Proteins using Low Molecular Weight Anionic Displacers
Authors:Amitava Kundu  Suresh Vunnum  Steven M. Cramer
Affiliation:(1) Howard P. Isermann Department of Chemical Engineering, Rensselaer Polytechnic Institute, Troy, NY, 12180
Abstract:A major impediment to the implementation of displacement chromatography has been the lack of suitable displacer compounds. Recently, it has been shown that low molecular weight dendritic polymers, protected amino acids and antibiotics can be successfully employed for displacement purification in cation-exchange systems. In this paper, a variety of low molecular weight anionic displacers are identified for the resolution of a bovine beta-lactoglobulin mixture into two closely related forms (A and B). A Dynamic Affinity plot is employed to evaluate the affinity of these low molecular weight compounds under various displacement conditions. In contrast to large polyelectrolyte displacers, the efficacy of these low molecular weight displacers are shown to be dependent on displacer concentration. In fact, the Dynamic Affinity Plot qualitatively predicts the transition from a displacement to a desorption regime with these low molecular weight displacers. In addition to the fundamental interest generated by low molecular weight displacers, it is likely that these displacers will have significant operational advantages as compared to large polyelectrolyte displacers. Furthermore, the ability to carry out selective displacement chromatography with these low molecular weight displacers offers significant potential for developing robust large scale displacement processes.
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