Virus preparations have been used as vaccines for over two hundred years, and more recently for gene therapy platforms. For gene delivery, conventional viral purification methods such as density gradient ultracentrifugation and precipitation can be difficult to scale and may not meet the purity standards required for a therapeutic. Consequently, chromatography has emerged as an advanced tool to meet large-scale therapeutic virus purification requirements. Viruses bind to chromatographic resins by electrostatic interactions.
We have previously demonstrated mammalian virus purification with the mixed-mode CHT Ceramic Hydroxyapatite Media (Kurosawa et al. 2014). CHT Media are a group of mixed-mode calcium affinity/cation exchange supports. The high negative surface charge on many viruses allows them to bind tightly to CHT calcium sites, permitting significant purification with high (>80%) yields. Such tight binding has also been utilized to provide robust viral clearance, which can be used to ensure viral safety during biomanufacturing (ICH Expert Working Group 1999, Moritz 2005).
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CHT XT is the newest addition to the CHT family of media. It has been designed for superior physical robustness to ensure it can be used repeatedly over a large number of cycles. It maintains the unique separation properties of CHT for aggregate removal and monoclonal antibody purification. In order to test its virus purification capabilities, we performed single-step purifications of influenza and dengue viruses using CHT XT Media.