Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, 200237, People's Republic of China.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, People's Republic of China.
Mikrochim Acta. 2019 Aug 3;186(9):600. doi: 10.1007/s00604-019-3712-2.
A hydrophilic interaction liquid chromatography (HILIC) material with application in glycoproteomics was obtained by sequential deposition of polyethyleneimine (PEI) and hyaluronic acid (HA) on a negatively charged substrate by means of electrostatic self-assembly. This kind of surface modification endows the material with excellent hydrophilicity and warrants efficient glycopeptides enrichment. The feasibility of this enrichment was verified by using dendritic mesoporous silica nanoparticles (DMSNs) and magnetic graphene oxide (MagG) as negatively charged substrates for PEI and HA adhesion. The two final products (DMSNs@PEI@HA and MagG@PEI@HA) exhibit high enrichment selectivity (molar ratios of IgG and BSA digests = 1:500 and 1:1000), sensitivity (detection limit, 2 fmol/μL), recovery (>90%) and enrichment capacity (300 mg/g). When using DMSNs@PEI@HA, 419 N-glycopeptides derived from 105 glycoproteins were identified. When using MagG@PEI@HA, 376 N-glycopeptides derived from 102 glycoproteins were identified, both from a 2 μL serum sample. This is better than by methods described in previous reports. Graphical abstract Schematic representation of hydrophilic modification of negatively charged nanomaterial substrates by electrostatic self-assembly techniques to obtain hydrophilic interaction liquid chromatography (HILIC) materials for enrichment of N-glycopeptides.
通过静电自组装技术，将聚乙烯亚胺（PEI）和透明质酸（HA）顺序沉积在带负电荷的基质上，得到一种用于糖蛋白质组学的亲水性相互作用液相色谱（HILIC）材料。这种表面修饰使材料具有优异的亲水性，并保证了高效的糖肽富集。通过使用树枝状介孔硅纳米粒子（DMSNs）和磁性氧化石墨烯（MagG）作为带负电荷的基底来粘附 PEI 和 HA，验证了这种富集的可行性。两种最终产物（DMSNs@PEI@HA 和 MagG@PEI@HA）表现出高的富集选择性（IgG 和 BSA 消化物的摩尔比=1:500 和 1:1000）、灵敏度（检测限，2 fmol/μL）、回收率（>90%）和富集能力（300 mg/g）。使用 DMSNs@PEI@HA 时，从 105 种糖蛋白中鉴定出 419 个 N-糖肽。使用 MagG@PEI@HA 时，从 102 种糖蛋白中鉴定出 376 个 N-糖肽，均来自 2 μL 血清样品。这比以前报道的方法要好。
