Comparative glycomic profiling of isotopically permethylated N-glycans by liquid chromatography/electrospray ionization mass spectrometry.
Comparative glycomic profiling of isotopically permethylated N-glycans by liquid chromatography/electrospray ionization mass spectrometry.
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DOI：
10.1002/rcm.6512
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发表时间：
2013-04-30
影响因子：
2
通讯作者：
Mechref, Yehia
中科院分区：
文献类型：
--
作者：
Hu, Yunli;Desantos-Garcia, Janie L.;Mechref, Yehia
基于质谱的比较糖组学对于疾病生物标志物的发现是必不可少的。然而，开发可靠的量化方法仍然是一项具有挑战性的任务。我们在这里报告了一种同位素标记策略，采用稳定的同位素碘甲烷进行比较糖组学分析，通过LC-ESI-MS。从模型糖蛋白和血清样品中释放的N-聚糖用碘甲烷（“轻”）和碘甲烷-d1或-d3（“重”）试剂进行全甲基化。然后在LC-ESI-MS分析之前，将全甲基化样品以等体积混合。同位素全甲基化的N-聚糖的峰强度比（重/轻，H/L）几乎等于理论值。观察到的差异主要与“重”碘甲烷试剂（碘甲烷-d1或-d3）的纯度有关。数据表明该策略同时定量来自代表不同队列的生物样品的N-聚糖的有效性。因此，该策略在单个LC-ESI-MS分析中比较多个样品是有效的。探讨了这种策略用于定义代表不同食管疾病的血清样本中糖组学差异的潜力。来源于生物样品和糖蛋白的同位素全甲基化N-聚糖的LC-ESI-MS比较糖组学分析可靠地确定了与生物条件或糖蛋白表达相关的聚糖变化。作为生物学应用，该策略允许可靠地量化与不同食管疾病相关的糖组学变化，包括高度异型增生、巴雷特病和食管腺癌。
Mass spectrometry based comparative glycomics is essential for disease biomarker discovery. However, developing a reliable quantification method is still a challenging task. We here report an isotopic labeling strategy employing stable isotopic iodomethane for comparative glycomic profiling by LC-ESI-MS. N-Glycans released from model glycoproteins and blood serum samples were permethylated with iodomethane (“light”) and iodomethane-d1 or -d3 (“heavy”) reagents. Permethylated samples were then mixed at equal volumes prior to LC-ESI-MS analysis. Peak intensity ratios of N-glycans isotopically permethylated (Heavy/Light, H/L) were almost equal to the theoretical values. Observed differences were mainly related to the purity of “heavy” iodomethane reagents (iodomethane-d1 or -d3). The data suggested the efficacy of this strategy to simultaneously quantify N-glycans derived from biological samples representing different cohorts. Accordingly, this strategy is effective in comparing multiple samples in a single LC-ESI-MS analysis. The potential of this strategy for defining glycomic differences in blood serum samples representing different esophageal diseases was explored. LC-ESI-MS comparative glycomic profiling of isotopically permethylated N-glycans derived from biological samples and glycoproteins reliably defined glycan changes associated with biological conditions or glycoproteins expression. As a biological application, this strategy permitted the reliable quantification of glycomic changes associated with different esophageal diseases, including high grade dysplasia, Barrett’s disease and esophageal adenocarcinoma.
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DOI：
10.1158/1078-0432.ccr-07-5261
发表时间：
2009-03-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Goldman R;Ressom HW;Varghese RS;Goldman L;Bascug G;Loffredo CA;Abdel-Hamid M;Gouda I;Ezzat S;Kyselova Z;Mechref Y;Novotny MV
通讯作者：
Novotny MV
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
DOI：
10.1111/j.1432-1033.1995.607_2.x
发表时间：
1995-10-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
BROCKHAUSEN, I;YANG, JM;TAYLORPAPADIMITRIOU, J
通讯作者：
TAYLORPAPADIMITRIOU, J
影响因子：
3
作者：
Goetz, John A.;Mechref, Yehia;Novotny, Milos V.
通讯作者：
Novotny, Milos V.
影响因子：
2.9
作者：
Hu, Yunli;Mechref, Yehia
通讯作者：
Mechref, Yehia