Microfiltration and Ultrafiltration for Viral Vector Purification

Membrane filtration is an emerging field in the realm of bioprocessing. One application is the purification of viral vectors-a crucial production step in vaccines and gene therapy. Past methods of purification such as ultracentrifugation and chromatography are time intensive, expensive, and inefficient.¹ Membrane purification, on the other hand, offers a flexible, scalable, and efficient way to separate viral vectors from contaminants.² This research focused on the purification optimization of adeno associated-viruses (AAV’s) from cell debris and proteins through microfiltration and ultrafiltration membranes. 20 nm silica particles were used to model AAVs, bovine serum albumin (BSA) was used to model HEK293 protein, and HEK293 lysed cells were used to model cell debris in viral vector supernatant. It was found that high hydrophilicity, large volumetric porosity, and large pore size of microfiltration membranes resulted in higher flux and recovery of 20 nm silica particles. Ultrafiltration was also successful in rejecting BSA and studying the effects of surface fouling.

References

[1]Florea, M., et al., “High-efficiency purification of divergent AAV serotypes using AAVX affinity chromatography,” Molecular therapy Moethods & Clinical Development, 2023, vol.28, pp. 146-157.

[2]Zydney, A., “New developments in membranes for bioprocessing – A review,” Journal of Membrane Science, 2021, vol. 620, pp.1-16.

Acknowledgements

This work would not have been possible without the NSF EPSCoR REU Supplement grant, and the Engineered Bioactive Interfaces and Devices REU program at University of Kentucky.