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用N,N-二甲基六烷基溴化铵涂层的毛细管对重组促红细胞生长素及其酶解产物进行毛细管电泳和毛细管电泳-电喷雾质谱分析
Analysis of Recombinant Erythropoietin and Its Tryptic Digestby Capillary Electrophoresis and Capillary Electrophoresis- Electrospray Ionization-Mass Spectrometry Using Capillary Coated with 6,6-Ionene
【摘要】 采用毛细管电泳技术研究了重组促红细胞生长素(rhEPO)的分离问题。用N,N-二甲基六烷基溴化铵(6,6-ionene)涂层的毛细管测定了rhEPO中唾液酸的微多相性,同时采用毛细管电泳-质谱(CE-M S)联用技术在22m in内鉴定了rhEPO20段胰酶消化肽中的11段。该方法简单快捷,重现性好,可用于蛋白质一级结构的测定。
【Abstract】 The microheterogeneity of recombinant human erythropoietin (rhEPO) was analyzed by capillary electrophoresis (CE) using a capillary coated with 6,6-ionene. The applicability of a volatile electrolyte for fast analysis of tryptic fragments of rhEPO with online CE-electrospray ionization-mass spectrometry (ESI-MS) was investigated, resulting in a reproducible separation of eleven rhEPO tryptic fragments within 22 min under the following conditions: running buffer 300 mmol/L acetic acid-ammonium acetate (HAc-NH4Ac), pH 4.80, separation voltage -15 kV and capillary temperature 25 ℃. The proposed method is rapid and effective, and can be used for the structural analysis of related proteins.
【Key words】 capillary electrophoresis; capillary electrophoresis coupled with electrospray ionization-mass spectrometry; 6,6-ionene coating; recombinant human erythropoietin; tryptic digest; microheterogeneity; tryptic mapping;
- 【文献出处】 色谱 ,Chinese Journal of Chromatography , 编辑部邮箱 ,2005年05期
- 【分类号】O658.9
- 【被引频次】4
- 【下载频次】267