In the pharmaceutical industry the purification of proteins and other biomolecules is a key step in the production of biopharmaceuticals. For this purpose downstream processing relies on a range of commercially available chromatography media (resins), featuring a large diversity of base matrices and ligands. While base matrices based on silica, polymethacrylate, dextran and others are also used, beaded agarose continues to be considered a very suitable base matrix material for large-scale biomolecule purification. Central to chromatography media performance is the porous structure, particularly as the pore space has a direct influence on the solute transport properties of the media. At the same time, the pore space comprises a collection of pores of heterogeneous sizes and irregular geometries. Thus, the pore space is characterized by a pore size distribution (PSD), rather than homogenous pores of a particular size. While the latter is virtually unattainable, research focused on tuning the PSD could improve selectivity and transport properties of the chromatography media. Such research is, however, in the case of materials such as agarose and dextran, often hampered by the fact that the materials tend to shrink when dried, limiting the usefulness of common techniques for investigating the PSD, e.g. gas sorption and mercury porosimetry. Instead, inverse size exclusion chromatography (ISEC) is applied in order to probe the pore space of chromatography media. In this work, we present our latest results using ISEC, atomic force microscopy and other methods to study the pore space of agarose-based chromatography media.

On the pore space of agarose-based chromatography media

251st National Meeting (2016)

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