色谱

色谱

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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

于冰,丛海林,刘虎威,李元宗,刘锋

  

  1. The Key Lab of Bioorganic Chemistry and Molecular Engineering of Ministry of Education,
    College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China
  • 收稿日期:2005-05-08 修回日期:1900-01-01 出版日期:2005-09-30 发布日期:1988-03-25
  • 通讯作者: 刘虎威

Analysis of Recombinant Erythropoietin and Its Tryptic Digest by Capillary Electrophoresis and Capillary Electrophoresis-Electrospray Ionization-Mass Spectrometry Using Capillary Coated with 6,6-Ionene

YU Bing, CONG Hailin, LIU Huwei, LI Yuanzong, LIU Feng
  

  1. The Key Lab of Bioorganic Chemistry and Molecular Engineering of Ministry of Education,
    College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China
  • Received:2005-05-08 Revised:1900-01-01 Online:2005-09-30 Published:1988-03-25

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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-MS)联用技术在22 min内鉴定了rhEPO 20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。

关键词: N,N-二甲基六烷基溴化铵涂层, 毛细管电泳, 毛细管电泳-质谱联用, 酶解产物, 肽图 , 微多相性, 重组促红细胞生长素

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: microheterogeneity, recombinant human erythropoietin, tryptic digest, 6,6-ionene coating, capillary electrophoresis coupled with electrospray ionization-mass spectrometry, tryptic mapping , capillary electrophoresis