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学科主题: 物理化学
题名: Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma
作者: Chen, Rui1;  Tan, Yexiong2;  Wang, Min2;  Wang, Fangjun1;  Yao, Zhenzhen3;  Dong, Liwei2;  Ye, Mingliang1;  Wang, Hongyang2;  Zou, Hanfa1
通讯作者: 邹汉法
刊名: MOLECULAR & CELLULAR PROTEOMICS
发表日期: 2011-07-01
DOI: 10.1074/mcp.M110.006445
卷: 10, 期:7, 页:M110;006445
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1809
产权排名: 1,1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods
摘要: Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: A robust, reproducible, and high throughput method was developed for the relative quantitative analysis of glycoprotein abundances in human serum. Instead of quantifying glycoproteins by glycopeptides in conventional quantitative glycoproteomics, glycoproteins were quantified by nonglycosylated peptides derived from the glycoprotein digest, which consists of the capture of glycoproteins in serum samples and the release of nonglycopeptides by trypsin digestion of captured glycoproteins followed by two-dimensional liquid chromatography-tandem MS analysis of released peptides. Protein quantification was achieved by comparing the spectrum counts of identified nonglycosylated peptides of glycoproteins between different samples. This method was demonstrated to have almost the same specificity and sensitivity in glycoproteins quantification as capture at glycopeptides level. The differential abundance of proteins present at as low as nanogram per milliliter levels was quantified with high confidence. The established method was applied to the analysis of human serum samples from healthy people and patients with hepatocellular carcinoma (HCC) to screen differential glycoproteins in HCC. Thirty eight glycoproteins were found with substantial concentration changes between normal and HCC serum samples, including alpha-fetoprotein, the only clinically used marker for HCC diagnosis. The abundance changes of three glycoproteins, i.e. galectin-3 binding protein, insulin-like growth factor binding protein 3, and thrombospondin 1, which were associated with the development of HCC, were further confirmed by enzyme-linked immunosorbent assay. In conclusion, the developed method was an effective approach to quantitatively analyze glycoproteins in human serum and could be further applied in the biomarker discovery for HCC and other cancers. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.006445, 1-13, 2011.
关键词[WOS]: FACTOR-BINDING PROTEIN-3 ;  GROWTH-FACTOR-I ;  LECTIN AFFINITY-CHROMATOGRAPHY ;  SERUM-ALPHA-FETOPROTEIN ;  HUMAN PLASMA PROTEOME ;  MASS-SPECTROMETRY ;  BREAST-CANCER ;  THROMBOSPONDIN LEVELS ;  SHOTGUN PROTEOMICS ;  LIVER-CANCER
语种: 英语
WOS记录号: WOS:000292541500005
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115246
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 R&A Ctr, Dalian 116023, Peoples R China
2.Second Mil Med Univ, Int Cooperat Lab Signal Transduct, Eastern Hepatobiliary Surg Inst, Shanghai 200438, Peoples R China
3.Second Mil Med Univ, Dept Biochem & Mol Biol, Shanghai 200433, Peoples R China

Recommended Citation:
Chen, Rui,Tan, Yexiong,Wang, Min,et al. Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma[J]. MOLECULAR & CELLULAR PROTEOMICS,2011,10(7):M110;006445.
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