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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Zhang, Ying Peng, Ye Bin, Zhichao Wang, Huijie Lu, Haojie |
| Description | Author Affiliation: Zhang Y ( Shanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, PR China); Peng Y ( Department of Chemistry, Fudan University, Shanghai 200032, PR China.); Bin Z ( Department of Chemistry, Fudan University, Shanghai 200032, PR China.); Wang H ( Shanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, PR China.); Lu H ( Shanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, PR China) |
| Abstract | N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. Highly specific identification of N-glycome remains a challenge while its biological significance is acknowledged. The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. Therefore, the highly specific purification procedure becomes a crucial step prior to MS analysis of the N-glycome. Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). At last, the proposed method was successfully applied in the profiling of N-glycome in human serum, in which a total of 31 N-glycan masses were identified. |
| File Format | HTM / HTML |
| ISSN | 00032670 |
| Journal | Analytica Chimica Acta |
| Volume Number | 934 |
| e-ISSN | 18734324 |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-08-31 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry |
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