Na Dong H, Park Eun J, Youn Yu S, Moon Byung W, Jo Yeong W, Lee Sung H, Kim Won-Bae, Sohn Yeowon, Lee Kang C
Drug Targeting Laboratory, College of Pharmacy, SungKyunKwan University, Suwon City, Korea.
Electrophoresis. 2004 Feb;25(3):476-9. doi: 10.1002/elps.200305684.
Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) using a hydrophilic replaceable polymer network matrix was applied to characterize the polyethylene glycol(PEG)ylated interferon alpha (PEG-IFN). The SDS-CGE method resulted in a clearer resolution in both the PEG-IFN species and the native IFN species. The distribution profile of PEGylation determined by SDS-CGE was consistent with that obtained by SDS-polyacrylamide gel electrophoresis (PAGE) with Coomassie blue or barium iodide staining. The result was also compared using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. SDS-CGE was also useful for monitoring the PEGylation reaction to optimize the reaction conditions, such as reaction molar ratio. This study shows the potential of SDS-CGE as a new method for characterizing the PEGylated proteins with advantages of speed, minimal sample consumption and high resolution.
使用亲水性可置换聚合物网络基质的十二烷基硫酸钠-毛细管凝胶电泳(SDS-CGE)被用于表征聚乙二醇(PEG)化干扰素α(PEG-IFN)。SDS-CGE方法在PEG-IFN种类和天然IFN种类中都产生了更清晰的分辨率。通过SDS-CGE确定的聚乙二醇化分布图谱与通过考马斯亮蓝或碘化钡染色的SDS-聚丙烯酰胺凝胶电泳(PAGE)获得的图谱一致。还使用基质辅助激光解吸/电离飞行时间质谱对结果进行了比较。SDS-CGE对于监测聚乙二醇化反应以优化反应条件(如反应摩尔比)也很有用。本研究表明SDS-CGE作为一种表征聚乙二醇化蛋白质的新方法具有潜力,具有速度快、样品消耗少和分辨率高的优点。