Significance of Porosity and Pore Accessibility for the Selection of Ion Exchange Adsorbents for Chromatographic Purification of Macromolecules

Selection of adsorbent for the development of purification process for biomolecules is crucial due to the requirement of large number of binding sites and adsorption area. Considering this, porous structure with high charge density is selected as an adsorbent for macromolecule purification. Such selection may provide high static binding capacity but causes loss of separation performance due to improper porosity of adsorbent in comparison to solute sizes involved. To address this problem for the screening of adsorbent, this work reports adsorbent selection procedure on the basis of adsorbent pore diameter (d(p)), solute hydrodynamic dimensions (RH), and flow velocity in support of binding capacity. Towards that end, this study evaluated the pore accessibility performance of varying characteristics adsorbents using tracers like acetone, lysozyme, and bovine serum albumin (BSA) by designing nonbinding conditions. All screened adsorbents showed certain loss of total surface area depending on the solute dimensions and pore size. Sepharose type adsorbents showed accessible area loss up to 25% for lysozyme and 50% for BSA. Sepabeads type showed 30% loss, while macroporous UNO type showed only 7% loss of surface area for lysozyme. The study correlates accessibility with size ratio beta (d(p)/RH). The value of beta > 38 is found to be required for the accessibility of total pore area and optimum separation performance of ion exchangers investigated. Accessibility and beta provide useful information for the selection of suitable adsorbent for the purification of macromolecules.