Ceramic hydroxyapatite media have been shown to be an effective multimodal chromatographic support for the purification of a variety of biomolecules including monoclonal and polyclonal antibodies, isoproteins, antibody fragments, and DNA. Ceramic hydroxyapatite requires special considerations during process-scale chromatography column packing due to its high specific gravity and rapid settling rate. Ceramic hydroxyapatite media have a free-settling density of 35-125 cm/hr for 40 μm particles and 125-275 cm/hr for 80 μm particles. Additionally, ceramic hydroxyapatite media are sensitive to mechanical shear, which can fracture the particles and create fines.
Open column systems such as the GE BioProcess Glass (BPG) column are commonly used for pilot-scale protein purification because of their simple design and ease of use. To transfer media into an open column, remove the top piston from the column tube.
In this report, a procedure for packing CHT ceramic hydroxyapatite media (Bio-Rad Laboratories, Inc.) in a BPG open column system is described, and column evaluation results after five independent packing studies are shown.