title: Fast solid-phase extraction of N-linked glycopeptides by amine-functionalized mesoporous silica nanoparticles.
abstract: Selective enrichment is a crucial step before the mass spectrometric analysis of glycoproteins. A new approach using 3-aminopropyltriethoxysilane (APTES)-functionalized mesoporous silica materials (SBA-15) was reported to enrich the glycoproteins. Selective extraction of glycopeptides was achieved through coupling the oxidized glycan chains on the glycopeptides with the amine groups on SBA-15 through a reductive amination reaction, then the captured glycopeptides were detached from the SBA-15 for the following MS analysis using the enzyme PNGase F. Because the mesoporous material has a confinement effect, the efficiency of enrichment and enzymatic deglycosylation was improved dramatically. The coupling time was shortened from 4 hours to 1 hour, and the deglycosylation time was greatly shortened from 6 hours to 3 hours. This approach was successfully applied to profile the N-glycoproteome of human colorectal cancer serum. 84 N-linked glycosylation sites from 56 N-linked glycoproteins were identified from as little as 5 μL serum.
authors.affiliation: Shanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, P. R. China. ying@fudan.edu.cn luhaojie@fudan.edu.cn.
authors.forename: Weili
authors.initials: W
authors.lastname: Miao
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Cai
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Haojie
authors.initials: H
authors.lastname: Lu
mesh_terms: D000588:Amines / Q000737:chemistry*; D006020:Glycopeptides / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D009584:Nitrogen / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D052616:Solid Phase Extraction / Q000379:methods*; D013997:Time Factors
chemical_list: D000588:Amines; D006020:Glycopeptides; C509969:SBA-15; D012822:Silicon Dioxide; D009584:Nitrogen