Evaluation of multi-modal high salt binding ion exchange materials.
The performance and selectivity of novel cation and anion exchange multi-modal chromatographic materials were evaluated. Desorption profiles of 13 proteins possessing a range of properties (e.g. size, charge and hydrophobicity) were determined on the cation exchange materials. Batch experiments were carried out by loading individual proteins on each resin at low salt, and examining the desorption of the proteins during sequential washes with increasing salt concentrations. While all of the resins exhibited some binding of proteins at elevated salt concentrations, this effect was more pronounced on the resins with aromatic ligands as compared to the materials with aliphatic ligands. As expected, materials with higher ionic capacities exhibited higher binding at elevated salts. In addition, some proteins exhibited high binding at elevated salt concentrations to all of the resins. The combined effect of charge and other secondary interactions with these multi-modal chromatographic materials enables high salt binding of a range of proteins as well as unique selectivities for the recovery of certain classes of proteins. Since the anion exchange materials all exhibited high binding at elevated salt concentrations the work with these materials focused on a study of elution strategies to remove proteins from these aromatic based materials. After evaluating various elution protocols, a combined strategy of pH change and chaotropic salt were shown to minimize electrostatic and hydrophobic interactions and was found to be an effective elution strategy for this class of anion exchange materials using peanut lectin as a model protein.
Yang
Ting
T
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY, USA.
Malmquist
Gunnar
G
Johansson
Bo-Lennart
BL
Maloisel
Jean-Luc
JL
Cramer
Steven
S
D002852:Chromatography, Ion Exchange / Q000295:instrumentation*; D007475:Ion Exchange Resins; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism; D012492:Salts / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D007475:Ion Exchange Resins; D011506:Proteins; D012492:Salts