Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma

	Development of Glycoprotein Capture-Based Label-Free Method for the High-throughput Screening of Differential Glycoproteins in Hepatocellular Carcinoma
	Chen, Rui 1; Tan, Yexiong 2; Wang, Min 2; Wang, Fangjun 1; Yao, Zhenzhen 3; Dong, Liwei 2; Ye, Mingliang 1; Wang, Hongyang 2; Zou, Hanfa 1
刊名	molecular & cellular proteomics
	2011-07-01
卷号	10 期号:7 页码:m110;006445
ISSN号	1535-9476
通讯作者	邹汉法
产权排序	1,1
中文摘要	development of glycoprotein capture-based label-free method for the high-throughput screening of differential glycoproteins in hepatocellular carcinoma
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.
学科主题	物理化学
WOS标题词	science & technology ; life sciences & biomedicine
类目[WOS]	biochemical research methods
研究领域[WOS]	biochemistry & molecular biology
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
收录类别	SCI
语种	英语
WOS记录号	WOS:000292541500005
公开日期	2012-07-09
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/115246]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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
推荐引用方式 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.,...&Zou, Hanfa.(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.