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A Fully Automated System with Online Sample Loading, Isotope Dimethyl Labeling and Multidimensional Separation for High-Throughput Quantitative Proteome Analysis
Wang, Fangjun1; Chen, Rui1; Zhu, Jun1; Sun, Deguang2; Song, Chunxia1; Wu, Yifeng2; Ye, Mingliang1; Wang, Liming2; Zou, Hanfa1
Source PublicationANALYTICAL CHEMISTRY
2010-04-01
DOI10.1021/ac100075y
Volume82Issue:7Pages:3007-3015
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordDIFFERENCE GEL-ELECTROPHORESIS ; SPECTROMETRY-BASED PROTEOMICS ; CODED AFFINITY TAGS ; HEPATOCELLULAR-CARCINOMA ; MASS-SPECTROMETRY ; BIOMARKER DISCOVERY ; CELL-CULTURE ; AMINO-ACIDS ; PROTEINS ; IDENTIFICATION
AbstractMultidimensional separation is often applied for large-scale qualitative and quantitative proteome analysis. A fully automated system with integration of a reversed phase-strong cation exchange (RP-SCX) biphasic trap column into vented sample injection system was developed to realize online sample loading, isotope dimethyl labeling and online multidimensional separation of the proteome samples. Comparing to conventionally manual isotope labeling and off-line fractionation technologies, this system is fully automated and time-saving, which is benefit for improving the quantification reproducibility and accuracy. As phosphate SCX monolith was integrated into the biphasic trap column, high sample injection flow rate and high-resolution stepwise fractionation could be easily achieved. similar to 1000 proteins could be quantified in similar to 30 h proteome analysis, and the proteome coverage of quantitative analysis can be further greatly improved by prolong the multidimensional separation time. This system was applied to analyze the different protein expression level of HCC and normal human liver tissues. After three times replicated analysis, finally 94 up-regulated and 249 down-regulated (HCC/Normal) proteins were successfully obtained. These significantly regulated proteins are widely validated by both gene and proteins expression studies previously. Such as some enzymes involved in urea cycle, methylation cycle and fatty acids catabolism in liver were all observed down-regulated.
Language英语
WOS IDWOS:000276004000056
Citation statistics
Cited Times:55[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/141637
Collection中国科学院大连化学物理研究所
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Dalian Med Univ, Affiliated Hosp 1, Dalian 116027, Peoples R China
Recommended Citation
GB/T 7714
Wang, Fangjun,Chen, Rui,Zhu, Jun,et al. A Fully Automated System with Online Sample Loading, Isotope Dimethyl Labeling and Multidimensional Separation for High-Throughput Quantitative Proteome Analysis[J]. ANALYTICAL CHEMISTRY,2010,82(7):3007-3015.
APA Wang, Fangjun.,Chen, Rui.,Zhu, Jun.,Sun, Deguang.,Song, Chunxia.,...&Zou, Hanfa.(2010).A Fully Automated System with Online Sample Loading, Isotope Dimethyl Labeling and Multidimensional Separation for High-Throughput Quantitative Proteome Analysis.ANALYTICAL CHEMISTRY,82(7),3007-3015.
MLA Wang, Fangjun,et al."A Fully Automated System with Online Sample Loading, Isotope Dimethyl Labeling and Multidimensional Separation for High-Throughput Quantitative Proteome Analysis".ANALYTICAL CHEMISTRY 82.7(2010):3007-3015.
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