Untitled

Abstract

A monomethioninesulfoxide rHuIFN alpha-2b variant was found to be present in the rHuIFN alpha-2b bulk drug substance in solution. The Met(111) residue was identified as Met sulfoxide by comparative tryptic/V8 mapping and mass spectrometric analysis. Nevertheless, the oxidation of the Met(111) residue did not seem to have a detectable effect on the biological activity of the molecule.

Evaluation of recombinant human interferon α-2b structure and stability by in-gel tryptic digestion, H/D exchange and mass spectrometry

“…Content of ␣-helix was calculated to be 56%, which is in agreement with data found in literature for native rHuINF ␣-2b [1,2]. Purity of interferon ␣-2b was examined by gel-filtration and reversed-phase chromatography with UV absorption and fluorescence detection.…”

“…Since rHuINF ␣-2b contains 12 Gln, 4 Asn and 5 Met residues, deamidation and oxidation take a part in protein degradation by influencing on structural and activity changes. Both processes were suggested to explain presence of different isoforms [1,23]. In this work, detailed structural characterization was performed.…”

“…Interferons are cytokines with a wide spectrum of biological activities including antiviral effect, cell growth inhibition and immunomodulatory properties [1,2]. There are three main types of human interferons according to the initial producer cell and immunospecificities: ␣ (leucocytes), ␤ (fibroblasts) and ␥ (immune).…”

A simple and rapid HPLC method for quantitation of interferon-α2b in dosage forms and delivery systems

“…System suitability tests showed that the developed method is of appropriate separation efficiency and peak shape. A notable advantage of this method over the previously reported HPLC methods for IFN␣2b [13,[17][18][19] was its considerably shorter (at least two times) run-time, which is of a great importance in practice, particularly when a high number of samples are to be analyzed (e.g., in regulatory and industrial settings). The results of assay tests on IFN-␣2b products marketed in Iran were representative for an applicable and easy-to-use method.…”

“…In addition, these methods are capable of being used during the production/formulation processes for monitoring of minor structural as well as conformational variations occurred in protein structure which can lead to significant changes in biological activity of the drug [15]. Most of the chromatographic methods presented for IFN-␣2b analysis have gain limited attention in practical use because of a series of inherent problems including very long run-time [13,[16][17][18][19], the need for special columns and/or other equipments, high variations in responses and, finally, the high expertise needed for use and interpretation of the results obtained from these methods [13,[18][19][20][21][22]. Therefore, in this article we have developed and validated a simple and available gradient reversed-phase HPLC method with UV detection for quantitation of IFN-␣2b in parenteral dosage forms and delivery systems.…”