title: Zirconia layer coated mesoporous silica microspheres as HILIC SPE materials for selective glycopeptide enrichment.
abstract: Characterization of protein glycosylation requires highly specific methods for the enrichment of glycopeptides because of their sub-stoichiometric glycosylation-site occupancy. The hydrophilic affinity based strategy has attracted more attention, owing to its broad glycan specificity, good reproducibility, and compatibility with mass spectrometric (MS) analysis. Several polar matrices have emerged for hydrophilic interaction chromatography (HILIC) approaches, including sepharose, cellulose, ZIC-HILIC and titania. Here, we present the solid-phase extraction (SPE) utility of zirconia coated mesoporous silica (ZrO(2)/MPS) microspheres for glycopeptide isolation prior to MS analysis. The high specificity of this SPE approach was demonstrated by the enrichment of glycopeptides from the digests of model glycoproteins in HILIC mode. ZrO(2)/MPS microspheres show superior selectivity and glycosylation heterogeneity coverage for glycopeptide enrichment to conventional sepharose. Furthermore, digested mixtures of the phosphoprotein α-casein and IgG were also treated with ZrO(2)/MPS HILIC SPE materials, which exhibited that glycopeptides could be effectively enriched with interference from phosphorylated peptides.
authors: Key Lab of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, China.
authors: Huihui
authors: H
authors: Wan
authors: Jingyu
authors: J
authors: Yan
authors: Long
authors: L
authors: Yu
authors: Qianying
authors: Q
authors: Sheng
authors: Xiuli
authors: X
authors: Zhang
authors: Xingya
authors: X
authors: Xue
authors: Xiuling
authors: X
authors: Li
authors: Xinmiao
authors: X
authors: Liang
mesh_terms: D002364:Caseins / Q000032:analysis / Q000737:chemistry; D002845:Chromatography / Q000379:methods*; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000032:analysis / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008863:Microspheres; D015203:Reproducibility of Results; D012822:Silicon Dioxide; D052616:Solid Phase Extraction / Q000379:methods*; D015040:Zirconium
chemical_list: D002364:Caseins; D006020:Glycopeptides; D007074:Immunoglobulin G; D012822:Silicon Dioxide; D015040:Zirconium; C028541:zirconium oxide