title: Determinants of protein retention characteristics on cation-exchange adsorbents.
abstract: There are currently a large number of commercially available strong and weak cation-exchange adsorbents for preparative protein purification, typically prepared by coupling charged ligands to a mechanically rigid porous bead. Because of the diverse chemical nature of the base matrix (carbohydrate, synthetic polymer, inorganic) and the coupling and ligand chemistry, cation-exchange adsorbents from different suppliers can differ substantially in chemical surface properties and physical structure. The differences in chemical properties can be in ionic capacity, hydrophobicity, the presence of hydrogen bond donors/acceptors, and the nature of the charged functional groups. In order to probe the effects of these factors on protein affinity, the isocratic retention of a set of model proteins was examined on a set of cation-exchange adsorbents to obtain a quantitative assessment of retention differences between adsorbents. Two adsorbent factors were found to be the dominant determinants of overall protein retention: the anion type and the adsorbent pore size distribution. Protein retention on strong cation-exchangers was found to be greater than that on corresponding weak cation-exchangers. Protein retention was increased on adsorbents with pore size distributions that include significant amounts of pore space with dimensions similar to those of the protein solute.
authors.affiliation: Merck Research Laboratories, Sumneytown Rd., West Point, PA 19486, USA.
authors.forename: P
authors.initials: P
authors.lastname: DePhillips
authors.forename: A M
authors.initials: AM
authors.lastname: Lenhoff
mesh_terms: D000327:Adsorption; D002411:Cation Exchange Resins; D002852:Chromatography, Ion Exchange; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry*
chemical_list: D002411:Cation Exchange Resins; D011506:Proteins