Physicochemical and biological assays for quality control of biopharmaceuticals: interferon alpha-2 case study.
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.
Silva
Marta M C G
MM
National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. msilva@nibsc.ac.uk
Lamarre
Baptiste
B
Cerasoli
Eleonora
E
Rakowska
Paulina
P
Hills
Anna
A
Bailey
Marc J A
MJ
Wheeler
Jun X
JX
Burns
Chris J
CJ
Gaines-Das
Rose E
RE
Jones
Chris
C
Robinson
C Jane
CJ
D001681:Biological Assay / Q000379:methods*; D001688:Biological Products / Q000737:chemistry* / Q000378:metabolism; D049109:Cell Proliferation; D002845:Chromatography / Q000379:methods; D015195:Drug Design; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000032:analysis* / Q000378:metabolism; D008715:Methionine / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013048:Specimen Handling; D013696:Temperature
D001688:Biological Products; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D008715:Methionine; D010100:Oxygen