Oxidation of recombinant methionyl human granulocyte colony stimulating factor
| Autor: | Auke Bult, Ed Hop, J.Léon E. Reubsaet, Serge D. Scholten, Willy J. M. Underberg, Jos H. Beijnen, Jan Teeuwsen |
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| Rok vydání: | 1998 |
| Předmět: |
Filgrastim
Molecular Sequence Data Clinical Biochemistry Pharmaceutical Science Mass Spectrometry Micellar electrokinetic chromatography Analytical Chemistry law.invention chemistry.chemical_compound Residue (chemistry) Methionine law Granulocyte Colony-Stimulating Factor Drug Discovery Humans Amino Acid Sequence Cyanogen Bromide Hydrogen peroxide Chromatography High Pressure Liquid Spectroscopy Chromatography Chemistry Hydrogen Peroxide Hydrogen-Ion Concentration Recombinant Proteins In vitro Recombinant DNA Spectrophotometry Ultraviolet Cyanogen bromide Chemical stability Oxidation-Reduction |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis. 17:283-289 |
| ISSN: | 0731-7085 |
| DOI: | 10.1016/s0731-7085(97)00199-4 |
| Popis: | 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 k obs value for the oxidation process is constant. Above pH 7.5, k obs 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 Met 121 oxidized, while the dioxidation product has oxidized Met 121 and Met 126 residues. The trioxidation products are the proteins with Met 121 , Met 126 and Met 137 or Met 0 , Met 121 and Met 126 oxidized. |
| Databáze: | OpenAIRE |
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