Systematic development and optimization of a cIEF method for the charge variants analysis of moderately basic to basic monoclonal antibodies using the Analytical Quality by Design strategy

Key Points

• This research aims to develop and optimize a cIEF method for analyzing charge variants of moderately basic to basic monoclonal antibodies.
• Developed a capillary isoelectric focusing (cIEF) method for profiling mAbs' isoforms.
• Utilized Analytical Quality by Design (AQbD) and mixture design for systematic optimization.
• Assessed calibration R² and pI biases as responses for method optimization.
• Successfully profiled five moderately basic to basic monoclonal antibodies using cIEF.
• Achieved high calibration R² values indicating robustness of the method.
• Minimized pI biases during optimization, enhancing accuracy of analysis.