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学科主题: 物理化学
题名: Facile preparation of monolithic immobilized metal affinity chromatography capillary columns for selective enrichment of phosphopeptides
作者: Zhang, Liyuan1, 2;  Wang, Hui1, 2;  Liang, Zhen1;  Yang, Kaiguang1;  Zhang, Lihua1;  Zhang, Yukui1
通讯作者: 张丽华
关键词: Immobilized metal affinity chromatography ;  Monolithic column ;  Phosphopeptides ;  Ti(4+) ;  Zr(4+)
刊名: JOURNAL OF SEPARATION SCIENCE
发表日期: 2011-09-01
DOI: 10.1002/jssc.201100169
卷: 34, 期:16-17, 页:2122-2130
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 1,1
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
摘要: Facile preparation of monolithic immobilized metal affinity chromatography capillary columns for selective enrichment of phosphopeptides
研究领域[WOS]: Chemistry
英文摘要: The selective enrichment of phosphopeptides with good reproducibility is vital for the indepth study of the phosphoproteome. Herein, we presented a novel method to prepare monolithic Ti(4+) or Zr(4+) immobilized metal affinity chromatography (IMAC) columns. Since succinimide was of high reaction activity with nitrilotriacetic acid (NTA) than traditional epoxy group, through the reaction of succinimide group on the monolithic matrix with nitrilotriacetic acid, the preparation of monolithic IMAC columns became facile. By the developed new method, not only the time required for Ti(4+) or Zr(4+) immobilization could be shortened from 10 to 1 h, but also the RSD of obtained phosphopeptide peak area was below 13%, even with IMAC columns prepared in different batches (n = 3). With such monolithic Ti(4+)- and Zr(4+)-IMAC columns, ten and seven phosphopeptides were effectively identified from the digests of a mixture of beta-casein and BSA, even with the molar ratio as low as 1:100, respectively. The phosphopeptide recovery was over 73%, and the loading capacity was over 0.8 mu mol/mL. Compared with commercial IMAC beads, the monolithic IMAC columns provide outstanding reproducibility, good selectivity, large loading capacity, and high recovery, which demonstrates that such monolithic IMAC columns might facilitate the in-depth analysis of phospho-proteomes.
关键词[WOS]: MASS-SPECTROMETRY ;  PHOSPHOPROTEOME ANALYSIS ;  PROTEIN-PHOSPHORYLATION ;  MITOCHONDRIAL PHOSPHOPROTEOME ;  MICROSPHERES ;  PEPTIDES
语种: 英语
WOS记录号: WOS:000295189400020
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115304
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China

Recommended Citation:
Zhang, Liyuan,Wang, Hui,Liang, Zhen,et al. Facile preparation of monolithic immobilized metal affinity chromatography capillary columns for selective enrichment of phosphopeptides[J]. JOURNAL OF SEPARATION SCIENCE,2011,34(16-17):2122-2130.
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