Effects of resin ligand density on yield and impurity clearance in preparative cation exchange chromatography. I. Mechanistic evaluation

• Published in: Journal of Chromatography A Vol. 1225; pp. 62 - 69
• Main Authors: Fogle, Jace, Mohan, Nina, Cheung, Eric, Persson, Josefine
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 17.02.2012
• Subjects: Aggregates; Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - isolation & purification; Cation exchange chromatography; cation exchange resins; Cation exchanging; Cations - chemistry; Charge variants; CHO Cells; Chromatography; Chromatography, Ion Exchange - methods; Clearances; Cricetinae; Cricetulus; Density; High molecular weight variants; Host cell protein; Humans; Hydrophobic and Hydrophilic Interactions; Ion Exchange Resins - chemistry; Ligand density; Ligands; Molecular Weight; Monoclonal antibodies; Polymers; Proteins; Proteins - chemistry; purification methods; Resins; Charge variants; Aggregates; Host cell protein; Cation exchange chromatography; Ligand density; Monoclonal antibodies; High molecular weight variants
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2011.12.048

► Effects of resin ligand density on cation exchange chromatography evaluated. ► Experiments were performed in bind-and-elute mode using gradient elution. ► Ligand density did not affect clearance of high molecular weight variants. ► Ligand density did affect resolution of a basic charge variant. The effects of resin ligand density on cation exchange chromatography performance in preparative monoclonal antibody purification processes were evaluated. A set of agarose-based cation exchange resins spanning a relatively wide range of ligand densities was tested using three different monoclonal antibodies with unique impurity profiles. Experiments were performed in bind-and-elute mode using gradient elution at both intermediate protein loadings and near saturation capacity. Ligand density did not affect clearance of high molecular weight variants under any of the conditions tested; however, ligand density did affect resolution of a basic charge variant in one case and changed host cell protein clearance in another instance. In general, the results indicate that the relationships between ligand density, retention, and resolution are affected by both characteristic charge and protein surface charge distribution.