Analysis of charge heterogeneities in mAbs using imaged CE

Area of Science:

• Biopharmaceutical analysis
• Analytical chemistry
• Protein characterization

Background:

• Monoclonal antibodies (mAbs) are critical therapeutics.
• Accurate characterization of mAb charge variants is essential for quality control.
• Existing methods like IEF and ion exchange chromatography have limitations.

Purpose of the Study:

• To develop and validate an imaged capillary electrophoresis (iCE) method for mAb charge variant analysis.
• To demonstrate the method's capability to provide both pI and quantitative charge variant data.
• To assess the suitability of iCE for research and manufacturing quality control.

Main Methods:

• Development and validation of a generic iCE method.
• Testing the method on a diverse range of therapeutic mAbs.
• Evaluating method robustness and suitability for various applications.

Main Results:

• The iCE method successfully measured charge variants for multiple mAbs.
• It provided information comparable to IEF and ion exchange chromatography.
• The method demonstrated suitability for a wide range of charge compositions (10-70% acidic species) and pIs (6.9-9.6).
• The technique proved to be robust and validated for quality control.

Conclusions:

• The developed iCE method is a powerful tool for mAb charge variant analysis.
• It offers advantages in identification, separation, and quantitation over conventional methods.
• iCE is suitable for research, quality control, release testing, characterization, stability studies, and comparability studies.
• This technology is a strong candidate for a platform analytical method in mAb development.