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Decrease of dynamic range of proteins in human plasma by ampholine immobilized polymer microspheres
Deng, Nan1,2; Zhu, Guijie1; Chen, Yuanbo1,2; Wu, Qi1,2; Liang, Zhen1; Sui, Zhigang1; Zhang, Lihua1; Yang, Kaiguang1; Zhang, Yukui1
KeywordAmpholine Fractionation Human Plasma Protein Dynamic Range
Source PublicationANALYTICA CHIMICA ACTA
2014-05-15
DOI10.1016/j.aca.2014.04.004
Volume826Pages:43-50
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordMULTIDIMENSIONAL LIQUID-CHROMATOGRAPHY ; LECTIN AFFINITY-CHROMATOGRAPHY ; TANDEM MASS-SPECTROMETRY ; LIGAND LIBRARY TREATMENT ; HUMAN SERUM PROTEOME ; EXCHANGE CHROMATOGRAPHY ; BIOMARKER DISCOVERY ; GEL ELECTROPHORESIS ; PEPTIDE LIBRARIES ; REVERSED-PHASE
AbstractA 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.
Language英语
WOS IDWOS:000335452300006
Citation statistics
Cited Times:4[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/145916
Collection中国科学院大连化学物理研究所
Affiliation1.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
GB/T 7714
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.
APA Deng, Nan.,Zhu, Guijie.,Chen, Yuanbo.,Wu, Qi.,Liang, Zhen.,...&Zhang, Yukui.(2014).Decrease of dynamic range of proteins in human plasma by ampholine immobilized polymer microspheres.ANALYTICA CHIMICA ACTA,826,43-50.
MLA Deng, Nan,et al."Decrease of dynamic range of proteins in human plasma by ampholine immobilized polymer microspheres".ANALYTICA CHIMICA ACTA 826(2014):43-50.
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