(415f) The Development of Cellulose Monolith with Anion Exchange/Pseudo-Affinity Functionalization for the Separation of Influenza Virus

The purification of virus or virus-like particles (VLP) from host cells such as mammalian cells and E. coli is a time consuming procedure and has been undergoing extensive study.  A monolith is a single-piece, interconnected, and porous stationary phase material which is widely applicable for separation technologies such as gas and liquid chromatography, high performance liquid chromatography, and capillary electro-chromatography [1, 2]. To make the virus separation technology faster and more efficiently, a new cellulose monolith with a tunable anion exchange/pseudo-affinity surface was developed. We propose a method to prepare cellulose fiber enhanced monolith for the separation of the influenza virus directly from mammalian cell culture media. The interconnected structure of the monolith was a result of the porogen generating pores and fiber interactions. To address the variance in expression of surface glycoproteins among different strains, as well as to maintain a specific separation, chromatography utilizing pseudo-affinity and anion-exchange immobilization was prepared for use with influenza virus. The cellulose monolith structure was characterized by Scanning Electron Microscopy. The two different modifications of anion-exchange and pseudo-affinity were quantified by element analysis. An H1N1 influenza virus strain (A/WSN/33) was replicated and harvested in MDCK cells and used directly for separation efficiency evaluation, after a freeze-thaw operation [3]. The result shows for the H1N1 strain from MDCK cell culture, the pseudo-affinity modification has higher virus separation efficiency.

Reference

[1] Potter OG, Hilder EF. Porous polymer monoliths for extraction: Diverse applications and platforms. Journal of separation science. 2008;31:1881-906.

[2] Namera A, Nakamoto A, Saito T, Miyazaki S. Monolith as a new sample preparation material: Recent devices and applications. Journal of separation science. 2011;34:901-24.

[3] Wolff MW, Reichl U. Downstream Processing: From Egg to Cell Culture‐Derived Influenza Virus Particles. Chemical engineering & technology. 2008;31:846-57.