This contribution reports on a study using Purexa™-MQ multimodal anion-exchange (AEX) membranes for protein polishing at elevated solution conductivities. Dynamic binding capacities (DBC 10 ) of bovine serum albumin (BSA), human immunoglobulins, and salmon sperm DNA (ss-DNA) are reported for various salt types, salt concentrations, flowrates, and pH. Using 1 mg/ml BSA, DBC 10 values for Purexa™-MQ were >90 mg/ml at conductivities up to 15 mS/cm. The membranes maintained a high, salt-tolerant BSA DBC 10 of 89.8 ± 2.7 (SD) over the course of 100 bind-elute cycles.Polishing studies with acidic and basic monoclonal antibodies at >2 kg/L loads showed that Purexa™-MQ had higher clearance of host cell proteins and aggregate species at high conductivity (13 mS/cm) and in the presence of phosphate than other commercial AEX media. Purexa™-MQ also had a high ss-DNA DBC 10 of 50 mg/ml at conductivities up to 15 mS/cm, markedly outperforming other commercial products.In addition to the effectiveness of Purexa™-MQ for protein polishing at elevated solution conductivities, its unusually high binding capacity for ss-DNA indicates potential applications for plasmid DNA purification.

Protein A chromatography is ubiquitous to antibody purification. The high specificity of Protein A for binding the Fc-region of antibodies and related products enables unmatched clearance of process impurities like host cell proteins, DNA, and virus particles. A recent development is the commercialization of research-scale Protein A membrane chromatography products that can perform capture step purification with short residence times (RT) on the order of seconds. This study investigates process-relevant performance and physical properties of four Protein A membranes: Purilogics Purexa™ PrA, Gore® Protein Capture Device, Cytiva HiTrap™ Fibro PrismA, and Sartorius Sartobind® Protein A. Performance metrics include dynamic binding capacity, equilibrium binding capacity, regeneration-reuse, impurity clearance, and elution volumes. Physical properties include permeability, pore diameter, specific surface area, and dead volume. Key results indicate that all membranes except the Gore® Protein Capture Device operate with flow rate-independent binding capacities; the Purilogics Purexa™ PrA and Cytiva HiTrap Fibro™ PrismA have binding capacities on par with resins, with orders of magnitude faster throughput; and dead volume and hydrodynamics play major roles in elution behavior. Results from this study will enable bioprocess scientists to understand the ways that Protein A membranes can fit into their antibody process development strategies.

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