title: A simple integrated system for rapid analysis of sialic-acid-containing N-glycopeptides from human serum.
abstract: Terminal sialylation is very important in cancer biology and has been extensively investigated for the discovery of potential clinical biomarkers of cancers. In this study, we presented a novel approach, by using of Ti(IV)-IMAC, to enrich sialic-acid-containing N-glycopeptides for the analysis of terminal sialylation. Compared with conventional method using TiO2 , this approach obtained 2.5 times more glycopeptides and glycosylation sites. Then, a simple integrated system combining filter-aided sample preparation, ACN-improved digestion, and Ti(IV)-IMAC enrichment was established for efficient analysis. In this system, protein digestion, glycopeptide enrichment, and deglycosylation were integrated and were performed sequentially in a single filter unit without any need for desalting, lyophilization, or sample transfer procedures. As a result, the number of identifications was improved by 1.5-fold and the total processing time was drastically reduced to only 7-8 h. By using this system, fast and efficient analysis of human serum sialylated N-glycoproteome was achieved. From only 1 μL of human serum, 217 unique glycopeptides and 194 glycosylation sites were successfully identified.
authors: CAS Key Lab of Separation Sciences for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China.
authors: Jun
authors: J
authors: Zhu
authors: Fangjun
authors: F
authors: Wang
authors: Kai
authors: K
authors: Cheng
authors: Jing
authors: J
authors: Dong
authors: Deguang
authors: D
authors: Sun
authors: Rui
authors: R
authors: Chen
authors: Liming
authors: L
authors: Wang
authors: Mingliang
authors: M
authors: Ye
authors: Hanfa
authors: H
authors: Zou
mesh_terms: D002853:Chromatography, Liquid; D006020:Glycopeptides / Q000097:blood* / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D019158:N-Acetylneuraminic Acid / Q000737:chemistry* / Q000378:metabolism; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods; D013997:Time Factors; D014025:Titanium / Q000737:chemistry*
chemical_list: D006020:Glycopeptides; C009495:titanium dioxide; D014025:Titanium; D019158:N-Acetylneuraminic Acid