A Novel Technology for Virus Vaccine Purification: Modeling and Operation of Continuous Annular Chromatography Unit

摘要

Recent decades have witnessed a large increase in the technological strategies for vaccine purification. Veterinary vaccines are also experiencing an upsurge in interest, especially for major problem areas such as foot and mouth disease (FMD), which is a devastating disease 'of live stock. Process economics and process intensification are the key drivers in order to develop a rational purification strategy for FMD vaccines. The continuous annular chromatography (CAC) is a potential and promising downstream process that allows large-scale continuous preparative chromatographic separation and purification of multi-component mixtures. Improvement of the purification of FMD vaccines via continuous annular chromatography (CAC) is the focus of this work. In chromatographic simulation and modeling studies, the complexity of the problem increases if the usually assumed equilibrium-dispersive condition, which neglects all transient resistances, is not invoked. In this work, the non-equilibrium model of Ozdural et al [1] is applied to continuous annular chromatography (CAC) modeling studies, so as to understand the contributions of mass transfer resistances on the elution profiles, and thereby providing the efficient and reliable simulation profiles. It was concluded that in order to reach a factual chromatographic column dynamics portrait the inclusion of non-equilibrium constraints in the modeling studies, as separate identities, is essential. In the experimental phase, inactivated and 300K Dalton tangential flow filtered BHK cell culture virus harvest, produced by SAP Institute (FMD vaccine production plant, Ankara, Turkey) is used. It was determined that tangential filtration was not efficient in removing host cell DNA from virus, despite that it is highly competent in regard of other proteins. The virus suspension is further treated with different types of resin filled columns so as to gain an insight of the chromatographic media to be used in CAC which was build upon our design and operation strategies are based on the non-equilibrium model predictions of the model developed in this study. It was demonstrated that careful selection of operation parameters is needed for an efficient separation of virus and host cell DNA, whereas the CAC model predictions might save considerable time for the optimization of chromatographic virus purification strategy. The significance of this work is threefold. First, it delineates the importance of the contributions of mass transfer resistances on the elution profiles of chromatographic separations. Second, use of this new methodology allows us to suggest protocols for system operation parameters and/or scale-up processes of CAC purification systems. Finally, the use of CAC emerged as a novel technology for continuous chromatographic virus purifications.