Separation And Enrichment Of Proteins By Using Different Functional Groups Modified Magnetic Nanoparticles And Analysis Of Proteomics

Proteomics studies have shown that low-abundance proteins?such as glycosylated proteins,thiolated proteins,phosphorylated proteins?are closely related to diseases and cancers.The separation and enrichment of these low abundance proteins from complex biological samples is the bottleneck of the study.In the first chapter,we briefly introduces several functionalized nanomaterials which is used for separating and enriching of glycosylated/phosphorylated proteins,and we focus on separating and enriching protein among them.In this paper,isolation and enrichment of glycoprotein,thiol-containing protein and phosphorylated protein were studied.First,the Oxime binding reaction was employed,we reported an approach to separate and enrich the glycoprotein/glycopeptide by Fe₃O₄ magnetic nanoparticles which containing the cleavable disulfide bond and functionalized by oxyamino groups reacted with aldehyde groups on the side chain of sialic acid oxide.Meanwhile,treating the Jurkat cells?acute T Cell leukemia cells?lysis in the same condition above to enrich the glycoprotein and analyzd by LC-MS/MS.?2?Through the thiol exchange reaction,the thiol-containing protein was separated and enriched by another magnetic nanoparticles which modified by pyridine disulfide bonds,which analyzed by SDS-PAGE and mass spectrometry.In additon,with the theory basis of?-elimination reaction,the phosphorylated proteins were separated and enriched by magnetic nanoparticles modified by pyridine disulfide bonds.At the end of this paper,mannose and galactose were used as starting materials,a kind of hexose consisting of four different glycosyl fragments was synthesized by selective protection and deprotection.Herein,three glycosyl fragments of the compound were synthesized,they are named?-D-mannopyranose substitutes,?-D-galactopyranose substitutes and?-D-furanose substitutes,respectively.