Degradation Pathways for Recombinant Human Macrophage Colony-Stimulating Factor in Aqueous Solution
作者： Jay A. Schrier , Richard A. Kenley , Rick Williams , Robert J. Corcoran , Yangkil Kim
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摘要: Recombinant human macrophage colony-stimulating factor (rhM-CSF) promotes proliferation and activity. rhM-CSF clinical trials are currently in progress require a stable, pharmaceutically acceptable dosage form. This report documents pH effects on degradation profiles aqueous solution, with an emphasis identifying products. Thus, highly purified was maintained at 30 to 50°C solutions adjusted 2 10. Stressed samples were analyzed by SDS-PAGE, reverse-phase HPLC, size exclusion scanning microcalorimetry, murine bone marrow The results show maximal protein stability the region 7 8. Degradation product chromatographic electrophoretic analyses distinctly different acidic versus alkaline solution. For stressed products isolated chromatographically electrophoretically. These characterized N-terminal amino acid sequencing, fast-atom bombardment mass spectrometry, peptide mapping. that major pathway solution involves cleavage two sites: aspartate169-proline170 aspartate213-proline214. A third potential site (aspartate45-proline46) remains intact under conditions cleave Asp169-Pro170 Asp213-Pro214. In proceeds via parallel intramolecular cross-linking reactions. β-elimination mechanism is proposed account for Consistent literature observations, retain biological
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