Monitoring polysorbate hydrolysis in therapeutic proteins using an ultrasensitive extraction-free fatty acid quantitation method
| Autor: | Ning Li, Caterina Riccardi, Douglas E. Kamen, Hui Xiao, Sisi Zhang |
|---|---|
| Rok vydání: | 2022 |
| Předmět: |
Biophysics
Polysorbates Fatty Acids Nonesterified Biochemistry Surface-Active Agents Hydrolysis chemistry.chemical_compound Enzymatic hydrolysis Denaturation (biochemistry) Lipase Molecular Biology Chromatography High Pressure Liquid chemistry.chemical_classification Polysorbate Chromatography biology Fatty Acids Antibodies Monoclonal Fatty acid Cell Biology Solubility chemistry biology.protein Degradation (geology) |
| Zdroj: | Analytical Biochemistry. 637:114472 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2021.114472 |
| Popis: | Polysorbates (PSs) are surfactants commonly added to therapeutic protein drug product formulations to protect proteins from denaturation and aggregation during storage, transportation, and delivery. However, enzymatic hydrolysis of PSs has been recognized as the primary route of PS degradation in monoclonal antibody formulations, resulting in the release of free fatty acids that drive undesired particulate formation. Here, we present a rapid lipase activity assay with optimized incubation conditions for accurate quantitation of free fatty acids without a fatty acid extraction step. This assay can detect low levels of PS degradation (0.000024% PS20 degradation) within 1 day with minimal sample preparation. The levels of released free fatty acids were found to strongly correlate with the degree of PS20 degradation. The case study described herein suggests that this approach can detect low levels of PS20 degradation caused by sub-ppm lipase levels within 1 day, compared with the duration of 14 days needed for PS degradation assays based on two-dimensional liquid chromatography-charge aerosol detection. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect