Glycopeptide / Glycoprotein Enriched The Study Of New Technologies

Glycosylation has been implicated in numerous biological processes including cell growth and developmental biology, immune response, tumor growth, metastasis, anticoagulation, cell to cell communication, and microbial pathogenesis. In this thesis aims, we explored new techniques to enrichment glycopeptides and glycoproteins, and those techniques were developed for the advancement of glycomics.The first chapter is the introduction of the development of glycomics, global methods for protein glycosylation analysis by mass spectrometry and recent development in the enrichment of glycopeptides for glycoproteomics.The second part centers on synthesis and application of aminobenzeneboronic acid capped magnetic carbonaceous (MC) microspheres material. Using this material to enrich glycopeptides in the tryptic digests of HRP and ASF, the results indicated that the particle could not only remarkably separate glycopeptides from peptide mixture but also effectively enrich glycopeptides. Furthermore, these nanoparticles can also effectively enrich glycoprotein from mixture of HRP and BSA.In the third part, an on-plate specific enrichment method is presented for the direct analysis of glycopeptides,4-Mercaptophenylboronic acid capped gold nanoparticles can be immobilized on plate using electric conduction paste, and then these spots were used to capture glycopeptides, non-glycopeptides will be washed away. Meanwhile, gold nanoparticles will develop the intensity of glycopeptides' peaks. This strategy greatly reduces sample loss and gets successful identification of glycopeptides.In the fourth part,4-Mercaptophenylboronic acid capped gold nanoparticles were used to identify the glycoproteins and their glycosylated sites for the liver of rat. A total of 46 unique N-glycosylation sites were detected in 46 different glycoproteins, and this supplied preliminary essay to introduce the material into the study of practical sample.