Comparative Evaluation of Commercial Protein A Membranes for the Rapid Purification of Antibodies

Abstract

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.

Keywords

• Protein A
• membrane chromatography
• dynamic binding capacity
• elution volume
• permeability
• bioprocessing industry