Designing new monoclonal antibody purification processes using mixed-mode chromatography sorbents

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 879; no. 13; pp. 836 - 843
• Main Authors: Toueille, Magali, Uzel, Audrey, Depoisier, Jean-François, Gantier, René
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.04.2011; Elsevier
• Subjects: adsorbents; Adsorption; Affinity; affinity chromatography; Analysis; Analytical, structural and metabolic biochemistry; Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - isolation & purification; Biological and medical sciences; CHO Cells; Chromatography; Chromatography, Affinity - instrumentation; Chromatography, Affinity - methods; Computational Biology; Cricetinae; Cricetulus; DoE; experimental design; Fundamental and applied biological sciences. Psychology; General pharmacology; High throughput screening; High-Throughput Screening Assays - instrumentation; High-Throughput Screening Assays - methods; Host cell proteins; hydrophobicity; manufacturing; Medical sciences; Mixed-mode; Monoclonal antibodies; Monoclonal antibody; Pharmacology. Drug treatments; Platforms; Protein A; proteins; Purification; purification methods; Resins; screening; Sorbents; Staphylococcal Protein A - chemistry; yields; High throughput screening; Host cell proteins; Mixed-mode; Purification; Monoclonal antibody; DoE; Mixed adsorbent; In vitro; Chromatography; Protein
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2011.02.047

Current platforms for purification of monoclonal antibodies, mostly relying on Protein A as a first capture step, are robust and efficient but significantly increase downstream purification costs, mainly due to Protein A resins. To decrease manufacturing costs, industry is increasingly considering the use of purification schemes without affinity Protein A resins. Mixed-mode chromatography can be used as a powerful alternative to standard purification platforms as it offers new selectivity and separation mechanisms exploiting a combination of both ionic and hydrophobic characteristics of antibodies and contaminating proteins. By using a design of experiments (DoE) approach and high throughput screening in 96-well plates, we developed four different two-steps MAb purification processes, based on the use of mixed-mode sorbents. Finally, three of the tested processes resulted in final purified Mab fractions containing less than 100 ppm of residual CHO proteins (CHOP), with overall process yields above 70%. These data show that mixed-mode chromatography sorbents, used at capture or intermediate purification steps, really expand the options of MAb purification process development with or without Protein A affinity chromatography.