Demand for improve the biomanufacturing process of gene therapy products has grown with the recent approval of several gene therapy based therapeutics. While the number of clinical trials investigating the therapeutic use of viral vectors continues to grow, the absence of an ideal scalable process limits both clinical advancement for many investigational products, as well as, successful commercialization of approved products. A major focus in development of gene therapy products is improvement of purification strategies for viral vectors. Currently, most viral vector downstream processes are based off methods developed for recombinant protein purification. While these methods are attractive for their scalability and routine usage in industry, challenges arise when applying these methods to much larger molecules, such as DNA and viruses. Column chromatography using porous particle-based media is one such protein purification technique, whose yield is low when adapted to gene therapy vector purification due to the heavy reliance on diffusive mass transport. This diffusive limitation, coupled with the fact that viral diffusion can be orders of magnitudes slower than proteins, prompts the need to investigate how porous particle morphology can be modified for improved viral absorption. Recent advances in computational modeling have allowed us to study and better understand the behavior of biological species in chromatographic separation all the way down to the bead/pore level, thereby empowering the simulation of different resin topology for improved viral absorption. This work examines the effect of chromatographic resin morphology on viral vector purification using computational fluid dynamics (CFD), scanning electron microscopy (SEM) and confocal laser microscopy. The structure characteristics for resins with different morphologies are determined using SEM. Resin morphology was varied though the use of chemically modified diatom silica frustules and compared with commercially available POROS 50 resin. In this presentation, CFD simulations will be provided for comparisons of flow patterns, peclet numbers, and tracer absorption for different resins. Comparison of experimental flow cell data for absorption kinetics provided a rational basis for CFD validation.

Investigation of biomimetic resin structure using CFD modeling of nano/meso fluid behavior for chromatographic gene therapy vector purification

255th National Meeting (2018)

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