Hydrazide Functionalized Core-Shell Magnetic Nanocomposites for Highly Specific Enrichment of N-Glycopeptides
Hydrazide Functionalized Core-Shell Magnetic Nanocomposites for Highly Specific Enrichment of N-Glycopeptides
复制标题
用于高度特异性富集 N-糖肽的酰肼功能化核壳磁性纳米复合材料
DOI：
10.1021/am501110e
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发表时间：
2014-05-28
影响因子：
9.5
通讯作者：
Lu, Haojie
中科院分区：
文献类型：
--
作者：
Liu, Liting;Yu, Meng;Lu, Haojie
鉴于糖基化对人类健康的生物学意义，来自复杂生物样品的糖蛋白质组分析高度倾向于发现疾病生物标志物和临床诊断。然而，由于与非糖基化肽相比，糖肽以相对低的丰度存在并且聚糖微异质性，糖肽需要从复杂的生物样品中高度选择性地富集以用于质谱分析。本文合成了一种新型的酰肼功能化核壳磁性纳米复合材料，用于高特异性富集N-糖肽。采用聚甲基丙烯酸（PMMA）对磁性核进行功能化，通过回流沉淀聚合法制备了磁性核和高分子壳上挂有高密度酰肼基团的Fe3O4@poly纳米复合材料（甲基丙烯酸）然后用己二酸二酰肼（ADH）对Fe3O4@PMAA进行表面改性，得到Fe3O4@poly（甲基丙烯酸酰肼）（Fe3O4@PMAH）。丰富的酰肼基团对糖肽的高度特异性富集和磁性核心使其适用于大规模，高通量和自动化的样品处理。此外，亲水性聚合物表面可以提供对其他肽的低非特异性吸附。与市售的酰肼树脂相比，Fe3O4@PMAH提高了标准糖肽的5倍以上的信噪比。最后，将该纳米复合材料应用于结直肠癌患者血清中N-糖蛋白质组的分析。在三个重复的实验中，总共鉴定了175个独特的糖肽和181个糖基化位点，对应于63个独特的糖蛋白，富集的糖肽和相应的糖蛋白的特异性分别为69.6%和80.9%。由于所有这些有吸引力的功能，我们相信，这种新的酰肼功能化的核壳磁性纳米复合材料将揭示新的光的轮廓复杂的生物样品中的N-糖蛋白质组的高通量。
In view of the biological significance of glycosylation for human health, profiling of glycoproteome from complex biological samples is highly inclined toward the discovery of disease biomarkers and clinical diagnosis. Nevertheless, because of the existence of glycopeptides at relatively low abundances compared with nonglycosylated peptides and glycan microheterogeneity, glycopeptides need to be highly selectively enriched from complex biological samples for mass spectrometry analysis. Herein, a new type of hydrazide functionalized core shell magnetic nanocomposite has been synthesized for highly specific enrichment of N-glycopeptides. The nanocomposites with both the magnetic core and the polymer shell hanging high density of hydrazide groups were prepared by first functionalization of the magnetic core with polymethacrylic acid by reflux precipitation polymerization to obtain the Fe3O4@poly(methacrylic acid) (Fe3O4@PMAA) and then modification of the surface of Fe3O4@PMAA with adipic acid dihydrazide (ADH) to obtain Fe3O4@poly(methacrylic hydrazide) (Fe3O4@PMAH). The abundant hydrazide groups toward highly specific enrichment of glycopeptides and the magnetic core make it suitable for large-scale, high-throughput, and automated sample processing. In addition, the hydrophilic polymer surface can provide low nonspecific adsorption of other peptides. Compared to commercially available hydrazide resin, Fe3O4@PMAH improved more than 5 times the signal-to-noise ratio of standard glycopeptides. Finally, this nanocomposite was applied in the profiling of N-glycoproteome from the colorectal cancer patient serum. In total, 175 unique glycopeptides and 181 glycosylation sites corresponding to 63 unique glycoproteins were identified in three repeated experiments, with the specificities of the enriched glycopeptides and corresponding glycoproteins of 69.6% and 80.9%, respectively. Because of all these attractive features, we believe that this novel hydrazide functionalized core shell magnetic nanocomposite will shed new light on the profiling of N-glycoproteome from complex biological samples in high throughput.