Separation of monoclonal antibody aggregates using a dry-compressed ceramic hydroxyapatite cuboid chromatography device

• Dry-compression packing of ceramic hydroxyapatite media in cuboid chromatography device • Fast, simple and reproducible packing technique • Precisely weighed mass of media within device • Fast and efficient protein separation • Efficient separation of monoclonal antibody aggregates Slurry packing techniques are commonly used for preparing ceramic hydroxyapatite (or CHT) multimodal chromatographic columns. These techniques, in addition to being dependent on operator expertise, can also result in significant porosity differences in the radial direction and packing density differences along the bed-height. If the dry CHT media could be directly packed into a chromatography device, some of the above problems associated with slurry packing could potentially be avoided. We discuss a dry-compression method for packing CHT (Type I) media in a cuboid chromatography device. The method is simple and reproducible, and does not require a high level of operator expertise. Also, very precise mass of media can be packed within a device using this method. Separation efficiency metrics such as the plate height and the reduced plate height obtained with a device packed using the proposed technique were found to be superior to those recommended by the manufacturer of the CHT media used in the current study. CHT columns have been shown to be suitable for the separation and analysis of monoclonal antibody samples. The suitability of using a dry-compressed CHT media packed cuboid chromatography device for separating aggregates from monoclonal antibody samples was also examined in this study.