Comparing MALDI-MS, RP-LC-MALDI-MS and RP-LC-ESI-MS glycomic profiles of permethylated N-glycans derived from model glycoproteins and human blood serum.
Comparing MALDI-MS, RP-LC-MALDI-MS and RP-LC-ESI-MS glycomic profiles of permethylated N-glycans derived from model glycoproteins and human blood serum.
复制标题
DOI：
10.1002/elps.201100703
复制
发表时间：
2012-07
期刊：
影响因子：
2.9
通讯作者：
Mechref, Yehia
中科院分区：
文献类型：
--
作者：
Hu, Yunli;Mechref, Yehia
纯化的糖蛋白和生物样品的糖组学分析通常通过不同的分析方法实现，但主要通过MS和LC-MS。增强的电离效率和改进的串联MS对全甲基化聚糖的解释促进了这种方法的普及。本研究的重点是使用MALDI-MS以及RP-LC-MALDI-MS和RP-LC-ESI-MS比较来自模型糖蛋白和人血清的全甲基化N-聚糖的糖组学谱。在模型糖蛋白的情况下，使用三种方法获得的糖组学谱非常相似。然而，在源自血清的聚糖的情况下，这并不完全正确。来自250 nl血清的还原和全甲基化N-聚糖的RP-LC-ESI-MS分析允许可靠地检测73种聚糖（其结构通过质量准确度和串联MS确认），而在相同样品的RP-LC-MALDI-MS和MALDI-MS分析中分别鉴定了53种和43种结构。RP-LC-ESI-MS分析有助于自动化和灵敏的串联MS采集。仅在RP-LC-ESI-MS分析中检测到的聚糖结构是以低丰度存在的聚糖。这表明RP-LC-ESI-MS分析的检测灵敏度更高，这是由于色谱分离促进了检测器的竞争性电离和饱和度降低。后者还允许分离几种结构异构体;然而，未检测到与连接有关的异构体分离。
The glycomic profiling of purified glycoproteins and biological specimen is routinely achieved through different analytical methods, but mainly through MS and LC-MS. The enhanced ionization efficiency and improved tandem MS interpretation of permethylated glycans have prompted the popularity of this approach. This study focuses on comparing the glycomic profiling of permethylated N-glycans derived from model glycoproteins and human blood serum using MALDI-MS as well as RP-LC-MALDI-MS and RP-LC-ESI-MS. In the case of model glycoproteins, the glycomic profiles acquired using the three methods were very comparable. However, this was not completely true in the case of glycans derived from blood serum. RP-LC-ESI-MS analysis of reduced and permethylated N-glycans derived from 250 nl of blood serum allowed the confident detection of 73 glycans (the structures of which were confirmed by mass accuracy and tandem MS), while 53 and 43 structures were identified in the case of RP-LC-MALDI-MS and MALDI-MS analyses of the same sample, respectively. RP-LC-ESI-MS analysis facilitates automated and sensitive tandem MS acquisitions. The glycan structures that were detected only in the RP-LC-ESI-MS analysis were glycans existing at low abundances. This is suggesting the higher detection sensitivity of RP-LC-ESI-MS analysis, originating from both reduced competitive ionization and saturation of detectors, facilitated by the chromatographic separation. The latter also permitted the separation of several structural isomers; however, isomeric separations pertaining to linkages were not detected.
登录
查看更多内容
影响因子：
7.4
作者：
Alley, William R., Jr.;Madera, Milan;Mechref, Yehia;Novotny, Milos V.
通讯作者：
Novotny, Milos V.
影响因子：
2
作者：
Delaney, J;Vouros, P
通讯作者：
Vouros, P
影响因子：
3
作者：
Goetz, John A.;Mechref, Yehia;Novotny, Milos V.
通讯作者：
Novotny, Milos V.
DOI：
10.1016/j.chroma.2008.09.105
发表时间：
2009-02-20
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Luo Q;Rejtar T;Wu SL;Karger BL
通讯作者：
Karger BL
影响因子：
4.3
作者：
Lam, Maggie P. Y.;Siu, S. O.;Lau, Edward;Mao, Xiuli;Sun, H. Z.;Chiu, Philip C. N.;Yeung, William S. B.;Cox, David M.;Chu, Ivan K.
通讯作者：
Chu, Ivan K.