Cellulose Monolith with Tunable Functionality for Immobilization of Influenza Virus

Advancements in influenza virus or virus like particle separation technology can greatly accelerate vaccine production. Due to frequent mutation of the influenza virus, rapid and effective separation of influenza virus directly from host cells still remains a challenge. In this paper, we developed a new cellulose fiber monolith-based chromatography with tunable anion exchange and pseudo-affinity modifications for the separation of cell cultureproduced influenza virus. A novel strategy is proposed for preparing cellulose fiber monolith, which enhances the mechanical property of the monolith. Controllable pore size and porosity were achieved by dispersing and removing a porogen into cellulose-dissolving solution. The structure of cellulose monolith was characterized by Scanning Electron Microscope and the porosity study, which confirmed the controllable average pore size of 10 micron and porosity up to 85%. Two virus separation modes including anion-exchange and pseudo-affinity were introduced into the cellulose monolith, subsequently. The two modifications were quantified by an Element Analyzer. A H1N1 influenza virus strain (A/WSN/33) was produced and harvested in adherent Madin-Darby Canine Kidney Epithelial (MDCK) cells and used directly for virus separation. The results indicated that the pseudo-affinity mode had higher virus separation efficiency and had less host cell dsDNA and protein contamination.