An all-membrane chromatographic purification process for monoclonal antibodies

Monoclonal antibodies (mAbs) are recombinant therapeutic proteins, produced predominantly by genetically engineered mammalian cells such as Chinese Hamster Ovary (CHO) cells. mAbs are purified through standardized chromatography platforms where Protein A resin is commercially ubiquitous as the first capture step. Packed-bed chromatography has well-known drawbacks, including high cost and low productivity due to extended residence times needed to overcome intraparticle mass transfer diffusional resistances. This research aims to develop a non-Protein A purification process for mAbs using nonwoven membrane adsorbers. Three nonwoven membrnaes were prepared using polyglycidyl methacrylate UV-grafted polyamide 11 membranes as the base material, each functionalized with a specific ligand: a multimodal anion exchange membrane (MMA-BMEA) produced by coupling N-Benzyl-N-methylethanolamine (BMEA), a multimodal cation exchange membrane (MMC-MPCA) functionalized with 2-mercaptopyridine-3-carboxylic acid (MPCA), and a strong anion exchange membrane (AEX-TEA) functionalized with triethylamine (TEA). The multimodal membranes showed remarkable protein-binding performance compared to commercial chromatographic resins under non-competitive and competitive conditions, when tested for direct capture in bind-and-elute mode on a CHO cell culture harvest containing therapeutic a monoclonal antibody (IgG1). A three-step all-membrane chromatographic process for mAb purification from CHO supernatant was then designed and compared with a standard resin-based purification process for the same mAb. While the all-membrane process fell slightly short of the stringent purity requirements for mAb products and a lower final product purity was obtained compared to the resin process, it demonstrated significant reduction in processing time and increased productivity, highlighting the potential of this technology and the need for further optimization to meet commercial standards.