DICP OpenIR
Subject Area物理化学
Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma
Chen, Rui1; Tan, Yexiong2; Wang, Min2; Wang, Fangjun1; Yao, Zhenzhen3; Dong, Liwei2; Ye, Mingliang1; Wang, Hongyang2; Zou, Hanfa1; Zou HF(邹汉法)
Source PublicationMOLECULAR & CELLULAR PROTEOMICS
2011-07-01
ISSN1535-9476
DOI10.1074/mcp.M110.006445
Volume10Issue:7Pages:M110;006445
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank1,1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordFACTOR-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
AbstractDevelopment of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma; 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.
Language英语
WOS IDWOS:000292541500005
Citation statistics
Cited Times:48[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/115246
Collection中国科学院大连化学物理研究所
Corresponding AuthorZou HF(邹汉法)
Affiliation1.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
GB/T 7714
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.
APA Chen, Rui.,Tan, Yexiong.,Wang, Min.,Wang, Fangjun.,Yao, Zhenzhen.,...&邹汉法.(2011).Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma.MOLECULAR & CELLULAR PROTEOMICS,10(7),M110;006445.
MLA Chen, Rui,et al."Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma".MOLECULAR & CELLULAR PROTEOMICS 10.7(2011):M110;006445.
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