Hydrodynamic Shear And Turbulent Flow Considerations For High Productivity Membrane Affinity Chromatography

By Eric Van Voorhees, Hasan Hashemisohi, Ph.D., W.L. Gore and Associates, Inc., Neha Saxena, Ph.D., Shunsuke Shiina, Ph.D., Rodrigo Gonzalez, Ph.D., AGC Biologics Inc.

Throughout the manufacturing process, therapeutic antibodies encounter various hydrodynamic forces and material contact surfaces that have the potential to affect protein stability and the likelihood of aggregation. The intensity of these forces can change as fluid flow rates and the elements of component design scale with batch size.

Membrane-based affinity chromatography devices are designed to offer shorter residence times and higher purification productivity in comparison to conventional packed-bed affinity resin columns. These features can lead to increased volumetric flow rates for a given column bed volume and its flow distribution components. Intuitively, two concerns can emerge regarding the forces exerted on proteins and their impact on quality: the effect of large fluid velocity gradients causing shear force, and the potential for turbulent flow regimes to introduce additional forces beyond those from mean flow.

This study aimed to identify hydrodynamic shear rate, cumulative shear stress, and Reynolds number values likely in high-productivity affinity membrane chromatography, based on size and scale. These values were benchmarked against those obtained from resin bead columns under comparable conditions. Moreover, the study involved experimentally purifying a clarified CHO cell harvest using various membrane and resin column size scales to compare profiles of low-order protein aggregation and fragmentation. The experiments also assessed whether large-scale aggregation at solid-phase interfaces during repeated cycles would significantly impact elution yields and column fouling.