色谱

色谱

• 研究简报 • 上一篇    下一篇

梯度加压毛细管电色谱分离蛋白质

赵良娟1,邹娟娟3,王秀丽3,阎超3,高如瑜2,刘明羽4,吕宪禹1

  

  1. 1. College of Life Science, Nankai University, Tianjin 300071, China; 2. State Key Laboratory of Elemento-Organic Chemistry, Nankai University, Tianjin 300071, China; 3. Unimicro (Shanghai) Technologies
    Co., Ltd., Shanghai 201203, China; 4. China Agricultural University, Beijing 100094, China
  • 收稿日期:2004-12-21 修回日期:1900-01-01 出版日期:2005-11-30 发布日期:1987-12-25
  • 通讯作者: 吕宪禹

Separation of Proteins by Gradient Pressurized Capillary Electrochromatography

ZHAO Liangjuan1, ZOU Juanjuan3, WANG Xiuli3, YAN Chao3, GAO Ruyu2, LIU Mingyu4, Lv Xianyu1   

  1. 1. College of Life Science, Nankai University, Tianjin 300071, China; 2. State Key Laboratory of Elemento-Organic Chemistry, Nankai University, Tianjin 300071, China; 3. Unimicro (Shanghai) Technologies
    Co., Ltd., Shanghai 201203, China; 4. China Agricultural University, Beijing 100094, China
  • Received:2004-12-21 Revised:1900-01-01 Online:2005-11-30 Published:1987-12-25

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摘要：

以1.5 μm无孔硅胶颗粒(non-porous silica,NPS)为固定相,采用电压和压力联合驱动流动相,用反相梯度加压毛细管电色谱(p-CEC)在7.5 min内实现了核糖核酸酶A、细胞色素C、溶菌酶和肌红蛋白等4种蛋白质的快速、高效的分离。比较了梯度加压毛细管电色谱和微柱液相色谱(μ-HPLC)分离蛋白质的结果,同时考察了固定相、离子对试剂三氟醋酸(TFA)浓度和电压等条件对梯度加压毛细管电色谱分离蛋白质的影响。结果表明,梯度p-CEC可以通过调节电压精细调节带电溶质的保留,提高分离选择性,缩短分离时间,得到较高的柱效。该方法在蛋白质分离分析及蛋白质组学的研究中具有很大的应用潜力，为高效快速地分离蛋白质开辟了新的途径。

关键词: 蛋白质 , 反相色谱, 加压毛细管电色谱, 梯度洗脱

Abstract:

The separation of four proteins including RNase A, cytochrome C, lysozyme and myoglobin was investigated by reversed-phase gradient pressurized capillary electrochromatography (p-CEC) with 1.5 μm non-porous silica C18 stationary phase. This mode was compared with micro-high performance liquid chromatography (μ-HPLC) and the effects of applied voltage, stationary phase and concentration of ion-pairing agent (trifluoroacetic acid, TFA) on the gradient p-CEC were also studied. This separation was performed rapidly on a new CEC instrument TrisepTM 2010 GV. The results showed that the retention mechanism of proteins in p-CEC mode is based on both chromatographic partitioning and electrophoretic migration. The results also demonstrated that p-CEC may have great potential for fast and efficient separation of proteins.

Key words: pressurized capillary electrochromatography, proteins , reversed-phase chromatography, gradient elution