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作 者:鲁飞杰 许之珏 邹霞[1] 郭勋祥[1] 张延[1]
机构地区:[1]上海交通大学系统生物医学研究院,系统生物医学教育部重点实验室,上海200240
出 处:《上海交通大学学报(医学版)》2016年第10期1408-1414,共7页Journal of Shanghai Jiao tong University:Medical Science
基 金:国家重点基础研究发展计划(“973”计划)(2012CB822103)~~
摘 要:目的·合成具有亲水性结构的新型末端炔基修饰的磁性纳米颗粒(简称磁珠),使其高效、特异性地富集糖蛋白或糖肽。方法·利用化学合成法在磁珠表面连接一段具有多个亲水性基团的末端炔基多碳链,并利用叠氮单糖标记的荧光多肽验证合成的磁珠的富集效能。结果·荧光测定及质谱鉴定结果表明合成的磁珠能够特异性地富集叠氮糖肽,且能够从牛血清白蛋白的胰蛋白酶解产物与荧光糖肽的混合物(质量比1 000∶1)中特异性地富集糖肽且效能不变。结论·成功合成具有亲水性结构的磁珠,其能够高效、特异性地富集糖肽,为后续复杂样品中各类糖蛋白或糖肽的富集及糖基化位点的鉴定提供新方法。Objective·To effectively and specifically enrich glycopeptides or glycoproteins by synthesizing a novel type of terminal alkyne-functionalized magnetic beads with a hydrophilic structure.Methods·An alkynyl carbons chain with multiple hydrophilic groups was linked to the surface of magnetic beads using chemical synthesis and the enrichment effect of synthesized magnetic beads were verified by GalNAz labeled fluorescent peptide.Results·The results of fluorescence value detection and mass spectrometry showed that synthesized magnetic beads could specifically enrich glycopeptides and were able to enrich glycopeptides from the mixture of bovine serum albumin peptides and azido glycopeptides in a mass ratio of 1 000∶1 with the same efficiency.Conclusion·Magnetic beads with a hydrophilic structure that can efficiently and specifically enrich glycopeptides were successfully synthesized and can provide a novel method for enriching various glycopeptides or glycoproteins and identifying the sites of glycosylation in complex samples.
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