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Weak anion and cation exchange mixed-bed microcolumn for protein separation
Hou, Chunyan1,2; Yuan, Huiming1; Qiao, Xiaoqiang1,2; Liu, Jinxiang1,2; Shan, Yichu1; Zhang, Lihua1; Liang, Zhen1; Zhang, Yukui1
关键词Ion-exchange Chromatography Protein Separation Tryptic Digestion Weak Anion And Cation Exchange Mixed-bed
刊名JOURNAL OF SEPARATION SCIENCE
2010-11-01
DOI10.1002/jssc.201000440
33期:21页:3299-3303
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Physical Sciences
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]ION-EXCHANGE ; LIQUID-CHROMATOGRAPHY ; MASS-SPECTROMETRY ; PH ; SALT
英文摘要To separate proteins with a wide distribution of pIs under the conditions compatible to online tryptic digestion (with preferable pH = 8.0), weak anion and cation exchange chromatography (WAX/WCX) mixed-bed microcolumn has been developed. With a mixture of five proteins with pIs ranging from 4.2 to 11.4, the effect of WAX/WCX ratio on the separation performance was investigated, and an optimum packing ratio of 1:1 w/w was obtained. Moreover, the undesirable hydrophobic interaction between the proteins and the stationary phase was suppressed with 10% ACN v/v added in the mobile phases. Under the optimized conditions compatible to tryptic digestion, basic and acidic proteins were resolved simultaneously, with RSDs of relative retention time on six columns less than 6%, indicating the good resolution and packing reproducibility. Furthermore, one RPLC fraction of proteins extracted from rat middle brain and the whole protein mixture extracted from rat liver were analyzed, respectively. The results demonstrated better separation performance on WAX/WCX microcolumns than that on both weak anion exchange chromatography and weak cation exchange chromatography at pH similar to 8. We anticipate that WAX/WCX microcolumns are promising for the integration of protein separation and tryptic digestion aiming at high-throughput proteome study.
语种英语
WOS记录号WOS:000284230700004
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被引频次:4[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/141684
专题中国科学院大连化学物理研究所
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
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Hou, Chunyan,Yuan, Huiming,Qiao, Xiaoqiang,et al. Weak anion and cation exchange mixed-bed microcolumn for protein separation[J]. JOURNAL OF SEPARATION SCIENCE,2010,33(21):3299-3303.
APA Hou, Chunyan.,Yuan, Huiming.,Qiao, Xiaoqiang.,Liu, Jinxiang.,Shan, Yichu.,...&Zhang, Yukui.(2010).Weak anion and cation exchange mixed-bed microcolumn for protein separation.JOURNAL OF SEPARATION SCIENCE,33(21),3299-3303.
MLA Hou, Chunyan,et al."Weak anion and cation exchange mixed-bed microcolumn for protein separation".JOURNAL OF SEPARATION SCIENCE 33.21(2010):3299-3303.
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