Chip-based reversed-phase liquid chromatography-mass spectrometry of permethylated N-linked glycans: a potential methodology for cancer-biomarker discovery.
Chip-based reversed-phase liquid chromatography-mass spectrometry of permethylated N-linked glycans: a potential methodology for cancer-biomarker discovery.
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DOI：
10.1021/ac100131e
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发表时间：
2010-06-15
影响因子：
7.4
通讯作者：
Novotny, Milos V.
中科院分区：
文献类型：
--
作者：
Alley, William R., Jr.;Madera, Milan;Mechref, Yehia;Novotny, Milos V.
生物体液和组织中蛋白质糖基化的研究具有重要的医学意义，因为糖链结构的变化现在已经与许多疾病有关。血糖谱变化的量化在寻找疾病生物标志物方面正变得越来越重要。在这里，我们报告了一种高度重复性的组合，即糖蛋白衍生N-连接多糖的糖链样品制备/固相衍生化与随后的基于微芯片的分离和质谱分析(MS)测量的组合。在我们之前描述的O-连接的糖链与氨基-硼烷络合物的还原β消除以减少N-连接的结构之后，N-连接的阿迪醇结构在二甲基甲酰胺介质中被有效地甲基化，以避免MS测量中的伪影。反相微流控液相色谱(LC)的甲基化N-连接的寡糖糖醇拆分一些密切相关的结构为规则的保留增量，帮助他们的结构指定。优化的LC梯度，结合纳米级的MS，已经被应用于血清中N-连接的多糖的定量测量，区分乳腺癌患者和对照组。
The study of protein glycosylation in biological fluids and tissues has substantial medical importance, as changes in glycan structures have now been associated with a number of diseases. Quantification of glycomic-profile changes is becoming increasingly important in the search for disease biomarkers. Here, we report a highly reproducible combination of a glycomic sample preparation/solid-phase derivatization of glycoprotein-derived N-linked glycans with their subsequent microchip-based separation and mass-spectrometric (MS) measurements. Following our previously-described reductive β-elimination for O-linked glycans with ammonia-borane complex to reduce N-linked structures, the N-linked alditol structures are effectively methylated in dimethylformamide medium to avoid artefacts in MS measurements. Reversed-phase microfluidic liquid chromatography (LC) of methylated N-linked oligosaccharide alditols resolved some closely related structures into regular retention increments, aiding in their structural assignments. Optimized LC gradients, together with nanospray MS, have been applied here in the quantitative measurements of N-linked glycans in blood serum, distinguishing breast cancer patients from control individuals.
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DOI：
10.1016/s0021-9673(00)96972-3
发表时间：
1990-01-19
期刊：
JOURNAL OF CHROMATOGRAPHY
影响因子：
--
作者：
ALPERT, AJ
通讯作者：
ALPERT, AJ
影响因子：
--
作者：
de Leoz ML;An HJ;Kronewitter S;Kim J;Beecroft S;Vinall R;Miyamoto S;de Vere White R;Lam KS;Lebrilla C
通讯作者：
Lebrilla C
影响因子：
3.4
作者：
Hashii, N;Kawasaki, N;Hayakawa, T
通讯作者：
Hayakawa, T
影响因子：
2
作者：
Delaney, J;Vouros, P
通讯作者：
Vouros, P
影响因子：
1.7
作者：
CORRADINI, C;CORRADINI, D;BONN, GK
通讯作者：
BONN, GK