Hydrolytic polysorbate 20 degradation – Sensitive detection of free fatty acids in biopharmaceuticals via UPLC-QDa analytics with isolator column
| Autor: | Stefan Carle, Patrick Garidel, Daniel Lakatos, Julia Buske, Dirk-H. Evers, Frank Hämmerling |
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| Rok vydání: | 2021 |
| Předmět: |
Clinical Biochemistry
Polysorbates Fatty Acids Nonesterified Sensitivity and Specificity 030226 pharmacology & pharmacy 01 natural sciences Biochemistry High-performance liquid chromatography Analytical Chemistry 03 medical and health sciences Hydrolysis chemistry.chemical_compound 0302 clinical medicine Enzymatic hydrolysis Solubility Chromatography High Pressure Liquid Polysorbate Biological Products Chromatography 010401 analytical chemistry Reproducibility of Results Cell Biology General Medicine 0104 chemical sciences Label-free quantification chemistry Linear Models Polysorbate 20 |
| Zdroj: | Journal of Chromatography B. 1174:122717 |
| ISSN: | 1570-0232 |
| Popis: | The enzymatic hydrolysis of polysorbates, e.g. induced by specific host cell proteins in biologics, is a known risk factor regarding the potential particle formation in the product over time. One of the root causes for this observation is an increase in free fatty acids (FA) within the formulation, which indicates the need for convenient monitoring of FA release. This study presents a novel UPLC-QDa based method to evaluate the content of the FAs esterified to polysorbate 20 (PS20) after hydrolysis. The presented method is label-free, i.e. independent of elaborate fluorophore-labeling and able to directly measure the ionized FAs. Furthermore, the method allows the determination of released FAs as percentage of ester bond hydrolysis and as absolute concentration expressed in ng/mL. Additionally, we describe for the first time in FA analytics the application of an isolator column, to remove trace levels of FAs present in the eluents to improve the sensitivity of the method. Lastly, the capabilities of the newly developed method are proven in case studies with three different monoclonal antibodies, which display characteristic FA release patterns in PS20-containing formulations. In summary, we developed a reliable, sensitive method for FA quantification in biologics, which could also be used as a predictive tool, considering FA solubility, regarding the formation of particles. |
| Databáze: | OpenAIRE |
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