化工学报 ›› 2016, Vol. 67 ›› Issue (7): 2893-2900.DOI: 10.11949/j.issn.0438-1157.20160095

• 分离工程 • 上一篇    下一篇

三种混合模式色谱填料色谱性能的比较

董彬彬1, 夏海锋1,2   

  1. 1. 江南大学工业生物技术教育部重点实验室, 江苏 无锡 214122;
    2. 江南大学粮食发酵工艺与技术国家工程实验室, 江苏 无锡 214122
  • 收稿日期:2016-01-21 修回日期:2016-04-12 出版日期:2016-07-05 发布日期:2016-07-05
  • 通讯作者: 夏海锋
  • 基金资助:

    国家自然科学基金项目(21206054);江苏省自然科学基金项目(BK20151123);中央高校基本科研业务费专项资金(JUSRP51401A)。

Comparison of three silica resins in chromatographic property for mixed-mode chromatography

DONG Binbin1, XIA Haifeng1,2   

  1. 1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China;
    2. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, Jiangsu, China
  • Received:2016-01-21 Revised:2016-04-12 Online:2016-07-05 Published:2016-07-05
  • Supported by:

    supported by the National Natural Science Foundation of China (21206054), the Natural Science Foundation of Jiangsu Province (BK20151123)and the Fundamental Research Funds for the Central Universities (JUSRP51401A).

摘要:

以γ-缩水甘油醚氧丙基三甲氧基硅烷为中间活化剂,将3-氨基吡啶、2-巯基苯并咪唑和2-巯基-1-甲基咪唑3种杂环配基键合到粒径为5 μm、孔径为30 nm的球形硅胶上,制备3种高效疏水电荷诱导色谱填料。利用溶菌酶、牛血清白蛋白和免疫球蛋白G作为模型蛋白,考察其在3种不同配基色谱柱上的保留行为和分离行为。比较了3种不同性质的模型蛋白在不同色谱柱上的色谱性能的异同。发现以2-巯基-1-甲基咪唑为配基的色谱柱具有良好的pH控制色谱行为和良好的分离性能,初步验证了高效疏水电荷诱导色谱的分离模式及潜在应用。

关键词: 生物分离, 蛋白质, 高效液相色谱, 疏水电荷诱导色谱, 保留因子, 分离行为

Abstract:

Spherical silica gel with a diameter of 5 μm and pore size of 30 nm was activated by KH-560 and coupled with three kinds of ligands (3-aminopyridine, 2-mercapto benzimidazole and 2-mercapto-1-methylimidazole) to prepare the high performance hydrophobic charge-induction chromatography (HPHCIC) column. The chromatographic properties of three columns were compared and evaluated with three model proteins (Lysozyme, BSA and IgG). The column with 2-mercapto-1-methylimidazole as ligand displayed a proper chromatography and separation behavior which controlled by pH. This work demonstrated that the novel synthetic materials can be utilized as the media of high performance hydrophobic charge induction chromatography.

Key words: bioseparation, protein, high performance liquid chromatography, hydrophobic charge-induction chromatography, retention factor, separation behavior

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