Extended Analysis of mAbs Using Ion-Exchange Chromatography on the Alliance™ iS Bio HPLC System

Large molecule biologics continue to be a major focus in the pharmaceutical market. These biopharmaceutical drugs require rigorous testing and monitoring throughout the production lifecycle beginning with the manufacturing process and ending with lifetime monitoring for stability and efficacy. Ion-exchange chromatography (IEX) is a widely used technique for lysine charge variant monitoring of monoclonal antibodies (mAbs) via high-performance liquid chromatography (HPLC). Ion-exchange methods, which employ either a salt or pH gradient, can be challenging on the chromatographic hardware over time due to the high salt concentration or variable pH used in the mobile phase system.

In this study an Alliance iS Bio HPLC System is employed in a 30-day robustness test using a salt gradient IEX method. A 3-protein standard is used for system baselining each day, while the lysine charge variants of Infliximab are monitored daily. Repeatability of retention time, total area, and relative area are monitored during the study. The chromatographic hardware and column demonstrated consistent performance achieving relative standard deviations as low as 0.1% for retention time, 2.44% for total area, and 0.95% for relative area over the 30 days of testing.

Benefits
- Reproducible multi-day HPLC performance on the Alliance iS Bio HPLC System
- Stable and repeatable salt gradient delivery using quaternary blending
- Consistent results for Infliximab charge variants with sample cooling for biomolecules down to 6 °C