(650e) Development of Fiber Enhanced Dual-Modal Cellulose Monolith for Influenza Virus Separation

Pan, X., Texas A&M University
Karim, M. N., Texas A&M University
Ramkumar, S. S., The Institute of Environmental and Human Health, Texas Tech University
Jinka, S., 2. Texas Tech University, Institute Environmental & Human Health

Development of fiber enhanced dual-modal cellulose monolith for influenza virus separation

Xinghua Pan1, Sudheer Jinka2, Seshadri Ramkumar2, M. Nazmul.Karim1*,  Vinitkumar Singh2

  1. Artie McFerrin Department of Chemical Engineering, Texas A&M University
  2.  Institute Environmental & Human Health, Texas Tech University

   *Corresponding author, Tel: (979) 845-9806, E-mail: naz.karim@che.tamu.edu


       Influenza virus imposes a serve threat to humans and animals, and its frequent mutation challenges the vaccine immunoprophylaxis, which still remains the most efficient method for flu epidemic prevention [1]. Recent widely applied, virus-producing hosts such as mammalian cells and E. colihave been reported for faster vaccine production compared to conventional chicken egg hosts [2]. The purification of virus or virus-like particles (VLP) from host cells is still a time consuming procedure and has been undergoing extensive study [2-4]. 

       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 electrochromatography.  The separation can target small molecules and large complex biomolecules. The advantages of monoliths compared to traditional porous beads are their mechanical robustness, ease of in-situ preparation at low cost, no void volumes, pore size controllability, and easy of functionalization. The advantages of the monolith also include easy shaping into various modes such as capillaries, columns, micropipette tips, and microfluidic channels. Besides, a highly interconnected porous structure allows high hydraulic permeability and convective flow dynamics in the monolith, which allows for higher flux rates. 

      Cellulose is among the most abundant bioproduct in plants. Its excellent biodegradability, sustainability, and biocompatibility make it as one of the ideal candidates for medical and chemical applications. The organic solution used for processing cellulose for membranes could be recycled, making the whole process an environmentally friendly and petroleum-independent green process. Numerous hydroxyl groups on cellulose are easy to modify for different separation purposes. Cellulose-based materials have been commonly applied in separation technologies such as centrifugation and filtration membranes, ion-exchange chromatography, and affinity chromatography.

      The preparation of cellulose monolith regarding its preparation method, mechanical properties, and pore size control has not been exclusively reported yet. To advance the virus separation technology faster and more efficiently, we propose a novel method to prepare dual-modal cellulose fiber enhanced monoliths for the separation of the influenza virus directly from mammalian cell culture media. 

      The interconnected structure of the monolith was contributed both by the porogen generating pores and fiber interactions. To address the variance in expression of surface glycoproteins among different strains, as well as maintaining the specific separation, a dual-mode 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 freeze-thaw operation. The results suggested the novel cellulose monolith could be a promising candidate for influenza virus separation.


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