Ceramic Hydroxyapatite (CHT) has been widely used in the production of biopharmaceuticals.  This mixed-mode chromatography media has unique capability of resolving biomolecules that are not separable by alternative means, owing to the synergistic interactions from the chelation of the calcium ions and the cationic exchange of the phosphoryl groups in its matrix. Multiple process and product-related impurities, such as aggregates/fragments of drug molecules, host cell proteins and nucleic acids, endotoxins, and contaminant viruses, can be effectively removed in a single chromatography operation. The rigid spherical macroporous particles of CHT offer high permeability for biomolecules and a linear pressure-flow relationship irrespective of column diameter, which allows easy production scale-up with the robustness and consistency demanded by bioprocessing.<br/>We have investigated some critical attributes in packing process-scale columns. CHT media can be packed by axial movement of column piston or constant flow of mobile phase, and no significant difference in final column quality is observed in the speed range of 100 ~ 400 cm per hour. On the basis of our results, we recommend testing column efficiency at a mobile phase linear velocity the same as column packing speed. Movement of column after packing may lead to initial minor change of plate count and/or asymmetry. However, subsequent movement does not seem to cause additional column bed settling or performance change. Practical tips for packing CHT columns will be provided in the present study.<br/>

Process-scale ceramic hydroxyapatite columns: Packing methods, column efficiency evaluation, and post-packing handling

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.