一种基于聚酰胺-胺型树枝状聚合物富集糖肽的新策略

Amine-Based Soluble Nanopolymers for Highly Effective Glycopeptide Enrichment

目的:提出了一种新型的糖肽富集试剂,将不同代数的含高密度N端的聚酰胺-胺(PAMAM)型树枝状聚合物固定到溴化氰活化的琼脂糖凝胶上用于糖肽的高效分离。方法:先用标准糖蛋白对试剂的富集条件进行优化,包括是否使用还原剂、不同酸度的结合溶液、不同洗脱液、不同试剂比例,将优化后的方法用于鼠脑裂解液糖肽的富集。结果与结论:将优化的方法用于鼠脑糖肽的富集,用第6代PAMAM鉴定到的糖肽数目是采用商业化酰肼材料的3倍,该试剂对糖肽富集的高选择性和高重现性为糖蛋白组学研究提供了新的工具。

Objective： Here, we presented a novel glycopeptides enrichment strategy based on the poly（amido- amine）（PAMAM） dendrimer reagent, which was functionalized with reactive groups for the site-specific capturing of glycopeptides. Methods： The high density amine group bearing PAMAM with different generation（PAMAM-G4, PAMAM-G6） were linked to CNBr-activated Sepharose bead, and then were used for isolation of glycopeptides from complex matrix. The method was optimized first with model glycoprotein, including reducing agents, binding solution, washing buffer, PAMAM to peptide ratio. Results ＆ Conclusion： This novel reagent demonstrated good reproducibility, high selectivity for enrichment of glycopeptides from complex mixture, which make it a useful tool for glycoproteomics researches.