Improvement of the Degradation Profiling of Eculizumab and Omalizumab Monoclonal Antibodies by Liquid Chromatography–Mass Spectrometry

AbstractThe control of the degradation of monoclonal antibodies is one of primary steps of the biologics development process. Traditionally a range of chromatographic and electrophoretic procedures is used for this purpose. However, the most comprehensive information about the degradation process can be obtained using a multiple attribute method (MAM) approach based on bottom-up liquid chromatography–tandem mass spectrometry (LC–MS/MS). We perform a comparative evaluation of the forced degradation of omalizumab and eculizumab monoclonal antibodies obtained by the bottom-up approach, isoelectric focusing, and high-performance liquid chromatography. An advantage of HPLC–MS/MS in terms of dynamic range, specificity, and flexibility in the evaluation of degraded protein forms was demonstrated. The results show that the developed bottom-up approach can be used in the studies of the degradation of monoclonal antibodies and the routine biopharmaceutical quality control.