色谱

色谱 ›› 2016, Vol. 34 ›› Issue (3): 232-239.DOI: 10.3724/SP.J.1123.2015.10030

• 研究论文 • 上一篇    下一篇

新型镍离子亲和色谱介质的制备及其性能考察

徐莉莉1, 陈革豫1, 张腾1, 张忠旗2, 韩广2, 李乾2, 赵金礼2, 刘建利1   

  1. 1. 西部资源生物与现代生物技术教育部重点实验室, 西北大学生命科学学院, 陕西 西安 710069;
    2. 西安华澳丽康活性蛋白与多肽工程中心, 陕西 西安 710054
  • 收稿日期:2015-10-20 出版日期:2016-03-08 发布日期:2012-09-28
  • 通讯作者: 刘建利
  • 基金资助:

    国家自然科学基金项目(30070905,20872118);陕西省重点实验室基金项目(2010JS097);陕西省教育厅基金项目(08jk477).

Preparation and properties of new nickel ion affinity chromatography medium

XU Lili1, CHEN Geyu1, ZHANG Teng1, ZHANG Zhongqi2, HAN Guang2, LI Qian2, ZHAO Jinli2, LIU Jianli1   

  1. 1. Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi'an 710069, China;
    2. Active Protein & Polypeptide Engineering Center of Xi'an Hua Ao Li Kang, Xi'an 710054, China
  • Received:2015-10-20 Online:2016-03-08 Published:2012-09-28
  • Supported by:

    National Natural Science Foundation of China (Nos. 30070905, 20872118); Foundation of Key Laboratory of Shaanxi Province (No. 2010JS097); Foundation of Shaanxi Provincial Department of Education (No. 08jk477).

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摘要：

以氯甲基化聚苯乙烯交联微球(氯球)为载体基质、N,N-二甲基甲酰胺(DMF)为溶剂、N,N-二异丙基乙胺(DIEA)催化,将合成的Lys(Boc)-OEt、氯乙酸乙酯连接到微球上。经水解使羧基裸露,与Ni2+螯合,制得新型镍离子亲和色谱介质。将其应用于His-Lys-Tyr三肽及Phe-His-Thr三肽纯化研究。以不含组氨酸的六肽作为杂质模型,用所制备的镍离子亲和介质进行分离。结果表明,所制备的镍离子亲和介质对两个含组氨酸的三肽具有很好的亲和作用,并实现了与不含组氨酸六肽的分离。

关键词: 含组氨酸短肽, 氯甲基化聚苯乙烯交联微球, 镍离子亲和色谱介质

Abstract:

With the chloromethylated crosslinking polystyrene microspheres (chlorine ball) as the matrices, N,N-dimethylformamide (DMF) as the solvent, activated by N,N-diisopropylethylamine (DIEA), the synthesized Lys(Boc)-OEt and ethyl chloroacetate were coupled to the matrices. After hydrolysis, the carboxyl group was released and chelated with Ni2+, and a new nickel ion affinity chromatographic medium was prepared. The affinity medium was applied to purify the His-Lys-Tyr and Phe-His-Thr tri-peptides. The results showed that the peptides containing histidine were retained on the affinity medium and separated with other substances.

Key words: chloromethylated crosslinking polystyrene microspheres, histidine-containing peptides, nickel ion affinity chromatography medium

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