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题名: Decrease of dynamic range of proteins in human plasma by ampholine immobilized polymer microspheres
作者: Deng, Nan1, 2;  Zhu, Guijie1;  Chen, Yuanbo1, 2;  Wu, Qi1, 2;  Liang, Zhen1;  Sui, Zhigang1;  Zhang, Lihua1;  Yang, Kaiguang1;  Zhang, Yukui1
关键词: Ampholine ;  Fractionation ;  Human plasma ;  Protein dynamic range
刊名: ANALYTICA CHIMICA ACTA
发表日期: 2014-05-15
DOI: 10.1016/j.aca.2014.04.004
卷: 826, 页:43-50
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
英文摘要: A novel protein sample pretreatment method based on ampholine immobilized polymer microsphere (ampholine@PM) was developed for the fractionation of intact proteins prior to protein digestion and peptide analysis to reduce the dynamic range of human plasma proteome. After incubation with our prepared ampholine@PM, the captured plasma proteins were successively desorbed by 2 M NaCl, 100 mM glycine-hydrochloric acid, and 30% (v/v) acetonitrile with 0.1% (v/v) trifluoroacetic acid. The SDS-PAGE results showed the protein dynamic range in such three fractions was obviously reduced as compared with the native plasma. On-particle digestion was ultimately performed to release all proteins retained on ampholine@PM. Followed by MuPIT analysis, the number of identified proteins in plasma was improved by 75% after ampholine@PM treatment. Furthermore, the spectral count of 9 high abundance proteins was decreased by 37.6-97.2%, and the identified low abundance protein (< 100 ng mL(-1)) number was increased from 4 to 17. These results demonstrated that the fractionation by ampholine@PM could efficiently decrease the protein dynamic range in abundance, beneficial to achieve the deep coverage identification of human plasma proteome. (c) 2014 Elsevier B. V. All rights reserved.
关键词[WOS]: MULTIDIMENSIONAL LIQUID-CHROMATOGRAPHY ;  LECTIN AFFINITY-CHROMATOGRAPHY ;  TANDEM MASS-SPECTROMETRY ;  LIGAND LIBRARY TREATMENT ;  HUMAN SERUM PROTEOME ;  EXCHANGE CHROMATOGRAPHY ;  BIOMARKER DISCOVERY ;  GEL ELECTROPHORESIS ;  PEPTIDE LIBRARIES ;  REVERSED-PHASE
语种: 英语
WOS记录号: WOS:000335452300006
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/145916
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, Key Lab Separat Sci Analyt Chem, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China

Recommended Citation:
Deng, Nan,Zhu, Guijie,Chen, Yuanbo,et al. Decrease of dynamic range of proteins in human plasma by ampholine immobilized polymer microspheres[J]. ANALYTICA CHIMICA ACTA,2014,826:43-50.
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