Comparison of alternative analytical techniques for the characterisation of the human serum proteome in HUPO Plasma Proteome Project

Li, Xiaohai; Gong, Yi; Wang, Ying; Wu, Songfeng; Cui, Yun; He, Ping; Zhang, Lu; Ying, Wantao; Zhang, Yangjun; Jiao, Liyan; He, Hui; Zhang, Zisen; He, Fuchu; Zhao, Xiulan; Qian, Xiaohong

doi:10.1002/9783527609482.ch10

Cited by 18 publications

(25 citation statements)

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“…Since, human plasma can be easily obtained, several laboratories have developed various proteomic strategies for the purpose of using plasma proteins as a source for possible disease-related biomarkers [7][8][9][10][11][12][13][14][15]22]. However, the great variety of protein types present, as well as the extended dynamic range of protein concentrations greatly hinders the identification of plasma biomarkers.…”

Section: Discussionmentioning

confidence: 99%

“…Although these depletion methods have shown good efficiency for albumin and IgG depletion, they often do not completely extract the targeted proteins or alternatively they nonspecifically bind some other plasma proteins. Recently, immunoaffinity methods have been developed to improve both the efficiency and the specificity for depletion of several abundant proteins in addition to albumin and IgGs in plasma or serum [7][8][9][10][11][12][13][14]. These chromatographic affinity methods enable specific depletion of 6-9 of the most abundant plasma proteins.…”

Section: Introductionmentioning

confidence: 99%

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Proteomic Analysis of Human Plasma Proteins by Two-Dimensional Gel Electrophoresis and by Antibody Arrays Following Depletion of High-Abundance Proteins

Desrosiers

Beaulieu

Buchanan

et al. 2007

Cell Biochem Biophys

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Detecting proteins that are present at lower levels in human plasma, for the identification of potential disease biomarkers, is complicated by a few highly abundant proteins. One promising strategy is the removal of these abundant proteins interfering with the analysis of plasma content by proteomic techniques. This study compared three affinity-based methods to remove the most abundant proteins in human plasma. Two of them, based on antibodies, which depletes the six or the 12 most abundant proteins, demonstrated the highest efficiency in enriching less abundant plasma proteins. Comparison of two anticoagulant treatments for plasma preparation, EDTA and CTAD, showed that this treatment influenced the patterns of lower-abundance proteins visible on 2-dimensional (2-D) gels. Several staining procedures including two fluorescent dyes, Sypro Ruby and Deep Purple, were also compared with a very sensitive silver staining method for the visualization of lower-abundance proteins on 2-D gels. Furthermore, treatments of lower-abundance plasma proteins with hydroxyethyl disulfide enhanced protein sharpness and resolution. The purpose of all these systematic comparisons was to select the most reliable methods in different steps of plasma preparation and handling as well as in analysis of proteins by 2-D gels to obtain highly reproducible patterns of lower-abundance plasma proteins. Importantly, the lower-abundance plasma proteins enriched by these optimized conditions were further analyzed by antibody microarrays allowing the identification of 61 proteins using 350 antibodies directed against signalling proteins suggesting that this proteomic strategy is a valuable approach for detecting potential plasma biomarkers.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Proteomic Analysis of Human Plasma Proteins by Two-Dimensional Gel Electrophoresis and by Antibody Arrays Following Depletion of High-Abundance Proteins

Desrosiers

Beaulieu

Buchanan

et al. 2007

Cell Biochem Biophys

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“…Qian et al compared five approaches for the characterization of human serum proteome [57]. Six proteins with the highest abundance were depleted and the serum was further analyzed using different combinations of separation methods such as chromatography/2D-PAGE, online or offline LC-LC-MS spectrum.…”

Section: Depletion Of High-abundance Proteinsmentioning

confidence: 99%

Recent advances in proteolysis and peptide/protein separation by chromatographic strategies

et al. 2010

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This review gives a broad glance on the progress of recent advances on proteolysis and peptide/protein separation by chromatographic strategies in the past ten years, covering the main research in these areas especially in China. The reviewed research focused on enzymatic micro-reactors and peptide separation in bottom-up approaches, and protein and peptide separation in top-down approaches. The new enzymatic micro-reactor is able to accelerate proteolytic reaction rate from conventionally a couple of hours to a few seconds, and the multiple dimensional chromatographic-separation with various models or arrays could sufficiently separate the proteomic mixture. These advances have significantly promoted the research of protein/peptide separation and identification in proteomics.proteolysis, enzymatic digestion, chromatographic separation, peptide enrichment, mass spectrometry

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“…More than leading HLPP, China has actively participated in HPPP [20], HBPP [21], Human Antibody Initiative (HAI) and Proteome Standard Initiative (PSI) [22].…”

Section: Contribution To Hupo and Hppmentioning

confidence: 99%

“…The proteomic technical platforms have been set up at international-level with high throughput, sensitivity, and resolution, which can be applied on large-scale and quantitative proteomics-based researches to speed up revealing human physiological and pathological mechanisms. Proteomes for some human organs and body fluids have been profiled since establishment of these platforms, such as protein expression profiles of human fetal liver [23] and adult liver [24,25], human plasma proteome [20,26], protein modification profile [27][28][29], comparative proteomics researches on cancer [30,31].…”

Section: Proteomics-based Technologiesmentioning

confidence: 99%

Proteomics in China: Ready for prime time

Gao

Zhang

Zheng

et al. 2010

Sci. China Life Sci.

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Proteomics is a newborn science focusing on the comprehensive systematic analysis of all proteins in the molecule machineries, organelles, cells, tissues, organs or intact organisms. It has been becoming one of the focuses in life sciences and the cutting-edged techniques in biotechnologies in the 21st century. During last decade, proteomics in China has developed much faster than other developing fields in life sciences. This review article briefly retrospects the origin and development of proteomics in China, also provides an overview on the representative scientific progress and perspectives.

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Comparison of alternative analytical techniques for the characterisation of the human serum proteome in HUPO Plasma Proteome Project

Cited by 18 publications

References 0 publications

Proteomic Analysis of Human Plasma Proteins by Two-Dimensional Gel Electrophoresis and by Antibody Arrays Following Depletion of High-Abundance Proteins

Proteomic Analysis of Human Plasma Proteins by Two-Dimensional Gel Electrophoresis and by Antibody Arrays Following Depletion of High-Abundance Proteins

Recent advances in proteolysis and peptide/protein separation by chromatographic strategies

Proteomics in China: Ready for prime time

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