Human serum N-glycome profiling via the newly developed asparagine immobilized cellulose/polymer nanohybrid.
Human serum N-glycome profiling via the newly developed asparagine immobilized cellulose/polymer nanohybrid.
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DOI：
10.1002/jssc.202200179
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发表时间：
2022-12
影响因子：
3.1
通讯作者：
Najam-ul-Haq, Muhammad
中科院分区：
文献类型：
--
作者：
Sajid, Muhammad Salman;Saleem, Muhammad Nakash;Jabeen, Fahmida;Saleem, Shafaq;Iqbal, Sabeen;Habib, Shahid;Ashiq, Muhammad Naeem;Ressom, Habtom W.;Najam-ul-Haq, Muhammad
关键词：
人血清n -链聚糖表达水平在疾病进展过程中发生变化。低丰度、结构多样性和共存的基质阻碍了它们在质谱分析中的检测。考虑到n -聚糖的亲水性，利用天冬酰胺功能化的氧环基团制备纤维素/聚合物（1,2-环氧-5-己烯）纳米杂化物，开发基于固相萃取的亲水性相互作用液相色谱吸附剂（纤维素/1,2-环氧-5-己烯/天冬酰胺）。通过扫描电子显微镜、能量色散x射线光谱和傅里叶变换红外光谱研究了其形貌、元素分析和表面性质。纤维素/聚合物纳米杂化物的大表面积（2.09×102 m2/g）有利于天冬酰胺的高密度固定，在优化条件下具有更好的亲水性相互作用。通过卵清蛋白和免疫球蛋白G释放的n -链聚糖来评估纳米杂交体/天冬酰胺的富集能力，其中分别检测到23个和13个n -链聚糖。该纳米杂化物对牛血清白蛋白的选择性为1:1200，灵敏度低至100阿托摩尔。n -聚糖的人血清分析鉴定了52个聚糖结构。这种新的富集策略在盐、蛋白质和内源性血清肽等存在的情况下富集血清n -链聚糖。
Human serum N-linked glycans expression levels change during the disease progression. The low-abundance, structural diversity and coexisting matrices hinder their detection in MS analysis. Considering the hydrophilic nature of N-glycans, cellulose/polymer (1,2-Epoxy-5-hexene) nanohybrid is fabricated with oxirane groups functionalized of asparagine to develop solid phase extraction based hydrophilic interaction liquid chromatography sorbent (cellulose/1,2-Epoxy-5-hexene/asparagine). The morphology, elemental analysis and surface properties are studied through scanning electron microscopy, energy dispersive x-ray spectroscopy, and Fourier-transform infrared spectroscopy. Large surface area of cellulose/polymer nanohybrid (2.09×102 m2/g) facilitates the high density of asparagine immobilization resulting in better hydrophilic interaction liquid chromatography enrichment under optimized conditions. The enrichment capability of nanohybrid/asparagine is assessed by the N-Linked glycans released from ovalbumin and immunoglobulin G where 23 and 13 N-glycans are detected respectively. The nanohybrid/asparagine shows selectivity of 1:1200 with spiked bovine serum albumin and sensitivity down to 100 attomole. Human serum profiling for N-glycans identifies 52 glycan structures. This new enrichment strategy enriches serum N-linked glycans in the presence of salts, proteins, and endogenous serum peptides, etc.
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