In this study carried out by Novartis, far-UV CD spectroscopy was qualified for the early development of a monoclonal antibody therapeutic. The high sensitivity of the Chirascan V100 enabled low-level detection of an IgG1 mAb in mixture with a structurally highly similar mAb. A demountable short-pathlength cell was used to minimize spectral contributions by formulation buffer excipients. Limit of Detection and Limit of Quantification were established and CD data contributed to a successful biotherapeutic submission to regulatory authorities.
Differences in secondary structure not significant*
Differences in tertiary structure are significant*
Monitor change under stressed conditions (forced degradation)
High sensitivity CD analysis of IgG1 samples subjected to a range of degradation conditions revealed minor differences in tertiary structure when compared to a control sample.
Quantifiable assessment of changes in tertiary structure
Statistical analysis enabled objective confirmation of spectral results. Degradation conditions have affected local environment of aromatic side chains (no changes were detected in secondary structure – results not shown).
Selected recent publications
Austerberry et al. 2017. “The Effect of Charge Mutations on the Stability and Aggregation of a Human Single Chain Fv Fragment.” European Journal of Pharmaceutics and Biopharmaceutics 115 (June): 18–30.