Integration of NMR Spectroscopy in an Analytical Workflow to Evaluate the Effects of Oxidative Stress on Abituzumab: Beyond the Fingerprint of mAbs.
Linda CerofoliniEnrico RaveraChristian FischerAndrea TrovatoFrancesca SaccoWolf PalinskyGabriella AngiuoniMarco FragaiFabio BaroniPublished in: Analytical chemistry (2023)
The assessment of the higher-order structure (HOS) by NMR is a powerful methodology to characterize the structural features of biologics. Forced oxidative stress studies are used to investigate the stability profile, to develop pharmaceutical formulations and analytical methods. Here, the effects of forced oxidative stress by H 2 O 2 on the monoclonal antibody Abituzumab have been characterized by a multianalytical approach combining NMR spectroscopy, mass spectrometry, differential scanning calorimetry, surface plasmon resonance, computational tools, and bioassays. This integrated strategy has provided qualitative and semiquantitative characterization of the samples and information at residue level of the effects that oxidation has on the HOS of Abituzumab, correlating them to the loss of the biological activity.
Keyphrases
- oxidative stress
- monoclonal antibody
- mass spectrometry
- liquid chromatography
- high resolution
- dna damage
- diabetic rats
- ischemia reperfusion injury
- induced apoptosis
- systematic review
- healthcare
- hydrogen peroxide
- nitric oxide
- gas chromatography
- tandem mass spectrometry
- capillary electrophoresis
- endoplasmic reticulum stress
- electron transfer