| Autor: |
Silva MM; National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. msilva@nibsc.ac.uk, Lamarre B, Cerasoli E, Rakowska P, Hills A, Bailey MJ, Wheeler JX, Burns CJ, Gaines-Das RE, Jones C, Robinson CJ |
| Jazyk: |
angličtina |
| Zdroj: |
Biologicals : journal of the International Association of Biological Standardization [Biologicals] 2008 Nov; Vol. 36 (6), pp. 383-92. Date of Electronic Publication: 2008 Aug 08. |
| DOI: |
10.1016/j.biologicals.2008.06.003 |
| Abstrakt: |
A selection of physicochemical and biological assays were investigated for their utility in detecting changes in preparations of Interferon alpha-2a and Interferon alpha-2b (IFN-alpha 2a, IFN-alpha 2b), which had been subjected to stressed conditions, in order to create models of biopharmaceutical products containing product-related impurities. The stress treatments, which included oxidation of methionine residues and storage at elevated temperatures for different periods of time, were designed to induce various degrees of degradation, aggregation or oxidation of the interferon. Biological activity of the stressed preparations was assessed in three different in vitro cell-based bioassay systems: a late-stage anti-proliferative assay and early-stage assays measuring reporter gene activation or endogenous gene expression by quantitative real time Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Relevant physicochemical methods such as SDS-PAGE, reverse phase (RP) chromatography, size-exclusion chromatography (SEC) and dynamic light scattering (DLS), proved their complementarity in detecting structural changes in the stressed preparations which were reflected by reductions in biological activity. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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