Simplifying Charge Heterogeneity Method Development with iCE3

This application notes discusses how charge heterogeneity analysis is crucial in biopharmaceutical development and quality control due to its impact on product stability and safety. Imaged capillary isoelectric focusing (icIEF) on iCE3 systems offers a streamlined method for measuring charge variants, reducing development timelines and enhancing product marketability. This method simplifies method development and ensures robustness through a straightforward workflow and optimization strategies.

Key Points

• icIEF Method Development:
  • Utilizes whole-column detection, eliminating the need for lengthy mobilization steps.
  • Assays are run in a cartridge with integrated electrolyte tanks, streamlining sample preparation.
  • Process from sample injection to capillary wash takes only about 10 minutes, cutting development timelines.
• Method Optimization:
  • Screening with a generic method provides separation profiles, followed by optimization with solubilizes and narrow pH-range ampholytes.
  • Method development from screening to optimization takes approximately 2.5 hours, ensuring efficiency.
  • iCE CFR software ensures compliance with regulatory standards for QC or GMP environments.
• Robust Method Considerations:
  • High salt concentrations can adversely affect resolution and reproducibility, remedied by sample dilution or buffer exchange.
  • Fresh reagents, particularly urea, are essential to maintain assay performance and avoid artificial alterations in protein composition.
  • With an optimized method, high-resolution cIEF separations enable effective characterization and monitoring of charge variants in various biopharmaceutical applications, including development, manufacturing, and formulations.

iCE3 has been discontinued as of June 2022. Consider Maurice, our next-generation iCE system, for charge heterogeneity analysis.