Membrane chromatography has emerged as alternative to packed bed chromatography due to its advantages of shorter cycle time, favorable mass transport, disposability, and potential to operate aseptically.  In addition, the macroporous structure of membranes is ideally suited for very large biomolecules such as plasmid DNA, virus-like particles, and live viruses, with mass transfer predominantly driven by convection.   In our work, we applied a 96-well high throughput (HT) workflow to the rapid development of a membrane chromatography capture step for large-virus purification.  The HT format was used to examine the effect of the feed matrix, mobile phase composition, and membrane chemistry on the capacity, yield, and impurity, allowing the multivariate parameter space to be thoroughly interrogated and conditions for optimal performance rapidly defined with a fraction of the material required at the lab scale.  The results from the HT format (0.7 cm2 membrane area) agree for the most part with results from the lab scale membrane (110 cm2). Some differences in the binding and elution behaviors are attributed, at least partly, to the differences in geometry and reduced flow uniformity of the 96-well format.

Development of membrane chromatography step for virus purification using high throughput membrane platform

247th National Meeting (2014)

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