Integration of NMR Spectroscopy in an Analytical Workflow to Evaluate the Effects of Oxidative Stress on Abituzumab: Beyond the Fingerprint of mAbs

The assessment of the higher-order structure (HOS) byNMR is apowerful methodology to characterize the structural features of biologics.Forced oxidative stress studies are used to investigate the stabilityprofile, to develop pharmaceutical formulations and analytical methods.Here, the effects of forced oxidative stress by H2O2 on the monoclonal antibody Abituzumab have been characterizedby a multianalytical approach combining NMR spectroscopy, mass spectrometry,differential scanning calorimetry, surface plasmon resonance, computationaltools, and bioassays. This integrated strategy has provided qualitativeand semiquantitative characterization of the samples and informationat residue level of the effects that oxidation has on the HOS of Abituzumab,correlating them to the loss of the biological activity.