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.

Preparation of Protein A Membrane Adsorbers Using Strain-Promoted, Copper-Free Dibenzocyclooctyne (DBCO)-Azide Click Chemistry

“…The apparent K d value also was benchmarked against other Protein A membranes. Previously, we measured K d values of 7.07 × 10 −2 ± 1.8 × 10 −2 mg/mL for Purilogics Purexa™ PrA and 7.63 × 10 −2 mg/mL for Sartorius Sartobind ® Protein A membranes [19]. K d values from Protein A resins have been reported to be between 4.5 × 10 −2 and 1.20 × 10 −1 mg/mL by Hahn et al [21].…”

“…Commercially, up until 2017, the only available Protein A membrane was the Sartorius Sartobind ® Protein A, which reported a DBC 10 of 5.9 mg IgG/mL membrane [18]. Newer commercial Protein A membranes, Purilogics' Purexa™ PrA and Cytiva's Fibro™ PrismA, were evaluated by Osuofa and Husson and shown to have ~70 mg/mL DBC 10 at 5 s residence time and ~90 mg/mL SBC for hIgG [19]. Sartorius' new convecdiff Protein A membranes have reported DBC 10 values of ~43 mg/mL at 12 s residence time [20].…”

Development of a New Affinity Gold Polymer Membrane with Immobilized Protein A

“…Commercially available bed-based chromatography products with Protein A, such as MabSelect™ by Cytiva, have dynamic binding capacities of 35-65 mg mL −1 and retention times (RT) of 2.4-6 min; Monofinity A Resin™ by Genscript achieved a DBC10 of 30 mg mL −1 with a RT of 4 min or even monoliths such as CIMac™ by BIA Separations of >5 mg mL −1 [6]. In a recent study by Osuofa and Husson [36], different new affinity membrane materials were tested regarding performance. The four tested membranes comprised a composite membrane by Gore, an electrospun fiber by Cytiva, an affinity membrane from Purilogics which is based on a convection-based flow, and the Sartobind ® Protein A membrane.…”

Continuous multi‐column capture of monoclonal antibodies with convective diffusive membrane adsorbers

“…At the same time, MAs have a large dead volume to stationary phase ratio (Matos et al, 2023), which leads to a broad residence time distribution due to high dispersion and back-mixing in the device (Ghosh et al, 2014;Hagemann et al, 2022;Madadkar et al, 2015). Thus, the elution volume of MAs increases with the dead volume to stationary phase ratio (Osuofa & Husson, 2023), resulting in a dilution of eluted product (Madadkar et al, 2015) and increased buffer consumption.…”