Effects of resin ligand density on yield and impurity clearance in preparative cation exchange chromatography. II. Process characterization

• Published in: Journal of Chromatography A Vol. 1225; pp. 70 - 78
• Main Authors: Fogle, Jace, Persson, Josefine
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 17.02.2012
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - isolation & purification; biopharmaceuticals; Cation exchange chromatography; CHO Cells; Chromatography; Chromatography, Ion Exchange - instrumentation; Chromatography, Ion Exchange - methods; Clearances; Cricetinae; Cricetulus; Density; Design of experiments; Elution; experimental design; High-throughput screening; Impurities; ion exchange resins; Ion Exchange Resins - chemistry; Ligand density; Ligands; Monoclonal antibodies; Multivariate Analysis; Proteins - chemistry; raw materials; Research Design; Resins; screening; Sepharose - chemistry; SP Sepharose™ Fast Flow; Specifications; Monoclonal antibodies; High-throughput screening; Cation exchange chromatography; Design of experiments; SP Sepharose™ Fast Flow; Ligand density
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2011.12.049

► Yield and impurity clearance evaluated using SP Sepharose™ Fast Flow. ► Ligand density varied over the commercial ligand density range. ► DoE study indicated no effect on yield, HCP clearance, aggregate clearance, or charge variants. ► Column experiments showed that ligand density may influence elution profiles. Ion exchange resins are key raw materials in biopharmaceutical manufacturing processes, and variability in ligand density has the potential to compromise process robustness if not controlled within appropriate ranges. In this study, yield and impurity clearance were evaluated for several preparative cation exchange chromatography steps using SP Sepharose™ Fast Flow resins at the high, low, and center of the commercial ligand density specification range. This was accomplished using a design of experiments (DoE) approach coupled to high-throughput screening in 96-well plate format, as well as column chromatography experiments with gradient elution. Results of the DoE study indicated that ligand density variation within the commercial specification range of SP Sepharose™ Fast Flow had no effect on yield, HCP clearance, aggregate clearance, or distribution of charge variants. However, results from the column experiments showed that ligand density has the potential to influence protein elution profiles which can lead to small changes in impurity clearance in some cases.