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Erschienen in:
01.12.2010 | Original Research Article
Physicochemical and Biologic Comparability of a Biosimilar Granulocyte Colony-Stimulating Factor with its Reference Product
Erschienen in: BioDrugs | Ausgabe 6/2010
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Background
Development of biosimilars requires physicochemical and biologic characterization to show comparability with a reference product. Zarzio® (filgrastim) is a biosimilar recombinant human granulocyte colony-stimulating factor (G-CSF) that has been approved in the EU using Neupogen® as its reference product.
Objective
The aim of this study was to compare the drug identity, purity, and bioactivity of Zarzio® (300 and 480μg/0.5mL solution) with Neupogen®, using a broad range of standard and advanced analytical methods.
Methods
Peptide mapping with UV detection and mass determination, circular dichroism (CD) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and liquid chromatography electrospray ionization (LC-ESI) mass spectrometry were among the analyses used to compare primary and higher-order protein structure. Cation-exchange chromatography (CEX) and reversed-phase high-performance liquid chromatography (RP-HPLC) were used to compare polarity and charge. Biologic characterization included comparison of G-CSF receptor binding affinity by surface plasmon resonance spectroscopy, an in vitro cell proliferation assay, and Western blot immunologic binding.
Results
The primary structures of Zarzio® and Neupogen® were shown to be identical by peptide mapping and other tests. CD and NMR spectroscopy demonstrated that the two products have comparable secondary and tertiary structures. RP-HPLC and other methods showed that the products have similar purity profiles. Comparable affinity with the G-CSF receptor GCSFR/CD114 was obtained using surface plasmon resonance spectroscopy, and comparable in vitro bioactivity was shown in a cell proliferation assay.
Conclusion
These results show the physicochemical and biologic comparability of Zarzio® and its reference product, Neupogen®.