DICP OpenIR
学科主题物理化学
Facile preparation of monolithic immobilized metal affinity chromatography capillary columns for selective enrichment of phosphopeptides
Zhang, Liyuan1,2; Wang, Hui1,2; Liang, Zhen1; Yang, Kaiguang1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
关键词Immobilized Metal Affinity Chromatography Monolithic Column Phosphopeptides Ti(4+) Zr(4+)
刊名JOURNAL OF SEPARATION SCIENCE
2011-09-01
ISSN待补充
DOI10.1002/jssc.201100169
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
研究领域[WOS]Chemistry
关键词[WOS]MASS-SPECTROMETRY ; PHOSPHOPROTEOME ANALYSIS ; PROTEIN-PHOSPHORYLATION ; MITOCHONDRIAL PHOSPHOPROTEOME ; MICROSPHERES ; PEPTIDES
英文摘要Facile preparation of monolithic immobilized metal affinity chromatography capillary columns for selective enrichment of phosphopeptides; 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.
语种英语
WOS记录号WOS:000295189400020
引用统计
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/115304
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
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
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GB/T 7714
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.
APA Zhang, Liyuan.,Wang, Hui.,Liang, Zhen.,Yang, Kaiguang.,Zhang, Lihua.,...&张丽华.(2011).Facile preparation of monolithic immobilized metal affinity chromatography capillary columns for selective enrichment of phosphopeptides.JOURNAL OF SEPARATION SCIENCE,34(16-17),2122-2130.
MLA Zhang, Liyuan,et al."Facile preparation of monolithic immobilized metal affinity chromatography capillary columns for selective enrichment of phosphopeptides".JOURNAL OF SEPARATION SCIENCE 34.16-17(2011):2122-2130.
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