Oxidation-Induced Destabilization of Model Antibody-Drug Conjugates
| Autor: | Matthias Winzer, Henning Gieseler, Jakob W. Buecheler, Christian Weber |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Immunoconjugates
Time Factors Drug Compounding Drug Storage Lysine Pharmaceutical Science 02 engineering and technology Protein aggregation 030226 pharmacology & pharmacy High-performance liquid chromatography Antioxidants Excipients 03 medical and health sciences 0302 clinical medicine Drug Stability Chromatography High Pressure Liquid chemistry.chemical_classification Reactive oxygen species Chromatography biology Protein Stability Antibodies Monoclonal Hydrogen Peroxide 021001 nanoscience & nanotechnology body regions chemistry Forced degradation biology.protein 0210 nano-technology Protein A Oxidation-Reduction Cysteine Conjugate |
| Zdroj: | Journal of pharmaceutical sciences. 108(3) |
| ISSN: | 1520-6017 |
| Popis: | Oxidation of biopharmaceutics represents a major degradation pathway, which may impact bioactivity, serum half-life, and colloidal stability. This study focused on the quantification of oxidation and its effects on structure and colloidal stability for a model antibody and its lysine (ADC-L) and cysteine (ADC-C) conjugates. The effects of oxidation were evaluated by a forced degradation study using H2O2 and a shelf-life simulation, which used degrading polysorbate 80 as source for reactive oxygen species. Differential scanning fluorimetry revealed decreasing transition temperatures of the CH2 domain with rising oxidation, resulting in a loss of colloidal stability as assessed by size-exclusion high pressure liquid chromatography. The conjugation technique influences structural changes of the monoclonal antibody (mAb) and subsequently alters the impact of oxidation. ADC-C was most effected by oxidation as the CH2 domain showed the biggest destabilization on conjugation compared to the mAb and ADC-L. Quantification of Fc methionine oxidation by analytical protein A chromatography revealed 4-fold higher oxidation after 8 weeks for the ADC-C compared to the mAb. Payload degradation was observed independently of the conjugation technique used or if free in solution by ultraviolet-visible. In addition, adding antioxidants can be a suitable approach to prevent oxidation and achieve baseline stabilization of the proteins. |
| Databáze: | OpenAIRE |
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