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Differential gel-based proteomic approach for cancer biomarker discovery using human plasma.

Authors
 Keun Na  ;  Min-Jung Lee  ;  Hye-Jin Jeong  ;  Hoguen Kim  ;  Young-Ki Paik 
Citation
 Methods in Molecular Biology (Clifton, N.J.), Vol.854 : 223-237, 2012 
Journal Title
Methods in Molecular Biology (Clifton, N.J.)
ISSN
 1064-3745 
Issue Date
2012
MeSH
Aged ; Biomarkers, Tumor/blood* ; Biomarkers, Tumor/isolation & purification* ; Biomarkers, Tumor/metabolism ; Carbocyanines/chemistry ; Carcinoma, Hepatocellular/blood ; Chromatography, Liquid ; Coloring Agents/chemistry ; Humans ; Hydrogen-Ion Concentration ; Image Processing, Computer-Assisted ; Liver Neoplasms/blood ; Male ; Nanotechnology ; Proteolysis ; Proteome/analysis* ; Proteome/isolation & purification* ; Proteome/metabolism ; Proteomics/methods* ; Staining and Labeling ; Tandem Mass Spectrometry ; Trypsin/metabolism ; Two-Dimensional Difference Gel Electrophoresis/methods*
Keywords
Two-dimensional fluorescence difference gel electrophoresis ; Narrow pH range ; Plasma proteomics ; Hepatocellular carcinoma ; Biomarker
Abstract
Two-dimensional fluorescence difference gel electrophoresis (2D DIGE) has become a general platform for analysis of various clinical samples such as biofluids and tissues. In comparison to conventional 2-D polyacrylamide gel electrophoresis (2D PAGE), 2D DIGE offers several advantages, such as accuracy and reproducibility between experiments, which facilitate spot-to-spot comparisons. Although whole plasma can be easily obtained, the complexity of plasma samples makes it challenging to analyze samples with good reproducibility. Here, we describe a method for decreasing protein complexity in plasma samples within a narrow pH range by depleting high-abundance plasma proteins. In combination with analysis of differentially expressed spots, trypsin digestion, identification of protein by mass spectrometry, and standard 2D PAGE and DIGE, this method has been optimized for comparison of plasma samples from healthy donors and patients diagnosed with hepatocellular carcinoma.
Full Text
http://link.springer.com/protocol/10.1007%2F978-1-61779-573-2_16
DOI
22311764
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Pathology (병리학교실) > 1. Journal Papers
Yonsei Authors
Kim, Hogeun(김호근)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/91455
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