Oxidation of recombinant methionyl human granulocyte colony stimulating factor

Journal of Pharmaceutical and Biomedical Analysis
J L ReubsaetW J Underberg

Abstract

The oxidation of methionine residues in recombinant methionyl human granulocyte colony stimulating factor with hydrogen peroxide has been investigated. Kinetic data of the oxidation were obtained by using reversed phase-high performance liquid chromatography. The stability-indicating capability of this system was confirmed with micellar electrokinetic capillary chromatography. In the pH range 1.9-7.5, the kobs value for the oxidation process is constant. Above pH 7.5, kobs tends to increase with increasing pH. In the pH range 1.9-11.8, four oxidation products were detected in RP-HPLC. Mass spectrometric analysis revealed that one mono-, one di- and two trioxidation products were formed. Using the cyanogen bromide cleavage method the nature of the oxidation products was determined. The mono-oxidation product is the protein with Met121 oxidized, while the dioxidation product has oxidized Met121 and Met126 residues. The trioxidation products are the proteins with Met121, Met126 and Met137 or Met0, Met121 and Met126 oxidized.

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Citations

Jul 9, 2013·Analytical and Bioanalytical Chemistry·Johann HolzmannHansjoerg Toll
Jul 20, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·W J UnderbergJ C Waterval
Dec 29, 2000·Journal of Pharmaceutical and Biomedical Analysis·H YokotaT Shibanuma
Sep 26, 2013·Pharmaceutical Research·Riccardo TorosantucciWim Jiskoot
Dec 11, 2014·Journal of Pharmaceutical Sciences·Tiina LipiäinenAnne Juppo
Jan 3, 2013·Journal of Pharmaceutical Sciences·Xian HuangRajesh Krishnamurthy
Dec 17, 2009·International Journal of Pharmaceutics·Renata PavisićMario Cindrić
Sep 3, 2009·Biotechnology and Bioengineering·Jared S BeeTheodore W Randolph
Jun 28, 2012·Journal of Controlled Release : Official Journal of the Controlled Release Society·Silvia ScaramuzzaPaolo Caliceti
Jan 22, 2013·Veterinary and Comparative Oncology·A K LeblancS A Kania
Feb 6, 2017·Pharmaceutical Research·Dmitry KryndushkinV Ashutosh Rao
Apr 27, 2021·Molecular Therapy. Methods & Clinical Development·Jared A DelmarXiaoyu Chen

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