Ampholine immobilized polymer microspheres for increasing coverage of human urinary proteome

被引:2
|
作者
Deng, Nan [1 ,2 ]
Chen, Yuanbo [2 ,3 ]
Liang, Zhen [2 ]
Bian, Yangyang [4 ]
Wang, Bing [1 ]
Sui, Zhigang [2 ]
Zhang, Xiaodan [2 ]
Yang, Kaiguang [2 ]
Zhang, Lihua [2 ]
Zhang, Yukui [2 ]
机构
[1] CNTC, Zhengzhou Tobacco Res Inst, Zhengzhou 450001, Peoples R China
[2] Chinese Acad Sci, Natl Chromatog Res & Anal Ctr, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
[3] Merieux Nutrisci China, Ningbo 315040, Peoples R China
[4] Zhengzhou Univ, Med Res Ctr, Affiliated Hosp 1, Zhengzhou 450052, Peoples R China
基金
中国国家自然科学基金;
关键词
Urine proteome; Sample treatment; Fractionation; Functionalized polymer microspheres; HYDROPHILIC INTERACTION CHROMATOGRAPHY; PROSTATE-CANCER; GLYCOSYLATION; PROTEINS; IDENTIFICATION; BIOMARKERS; CARRIER; KIDNEY; ACID; HEPARIN;
D O I
10.1016/j.talanta.2020.120931
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Urinary proteome, as an important component of body fluid proteome, could reflect kidney, urogenital tract function and pathological changes of human organs. This study reports a convenient strategy for urine proteome analysis through ampholine immobilized polymer microsphere (ampholine@PM) fractionation strategy. After ampholine@PM treatment, 16,543 unique peptides corresponding to 2173 non-redundant urinary proteins were identified, while only 856 proteins, corresponding to 3524 peptides were identified in the crude urine sample. The number of proteins and peptides was increased by 1.54 and 3.69 times, respectively. 31 urinary candidate biomarkers have also been identified (17 candidate biomarkers of glomerular injury and 14 candidate biomarkers of tubular injury), showing the potential of our strategy in urinary biomarker discovery study. In additional to the urine proteome, N-glycoproteome analysis was also performed after ampholine@PM fractionation followed by the N-glycopeptides enrichment. The number was increased from 144 to 281 for N-glycoproteins, 261 to 709 for N-glycopeptides, and 226 to 493 for N-glycosylation sites, after ampholine@PM treatment. Based on the significant increase on the identified N-glycoprotein number, ampholine@PM fractionation strategy also offered a beneficial tool for the post translational modification analysis of urine proteome.
引用
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页数:8
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