The ProPac® WAX-10 is a high-resolution weak anion-exchange column for the separation of protein isoforms. Ion-exchange columns provide characterization and quality control assessment of closely related protein variants. The rigid, nonporous pellicular resin provides exceptionally high resolving power, permitting the separation of proteins that differ by as little as one charged residue. The neutral hydrophilic pellicular coat eliminates protein-resin hydrophobic interactions, resulting in highly efficient peaks with excellent recovery. Selectivity differences between the ProPac WAX-10 and the ProPac SAX-10 provide flexibility in maximizing differences of closely related species.
The ProPac WAX-10 column is a weak anion exchanger with tertiary amine functional groups attached to a nonporous core. Traditionally, porous resins have been used for column packing materials because of the increased surface area provided by the pores. However, proteins, like other large molecules, are often too big to fit through these pores, which can result in significant peak shape deterioration through mass transfer restrictions. The ProPac resins are based on a nonporous polymeric particle coated with a proprietary hydrophilic layer and are functionalized with grafted linear, ion-exchange moieties. The nonporous core particle provides the high-efficiency separations, the hydrophilic layer prevents unwanted secondary interactions that can destroy a separation, and the grafted ion-exchange changins provide much higher capacity than is typical with nonporous resins. Differences in selectivity between the ProPac WAX-10 and ProPac SAX-10 provide flexibility in maximizing resolution of closely related species. The choice of whether to use a strong or weak anion or cation exchanger is somewhat empirical, and is based on the pI of the protein in question. As with all our ProPac columns, ProPac WAX-10 columns are manufactured and tested under the strictest specifications, resulting in unmatched column-to-column and lot-to-lot reproducibility. Dionex offers these columns as a lot select package; three columns from one lot or three columns each from a different lot, to assist the quality control scientist in the validation of their methods. The power of the ProPac WAX-10 has been demonstrated in the separation of the protein of interest from minor components, such as ovalbumin 1 from its minor components and carbonic anhydrase from its minor components.
|
