Automated measurement of permethylated serum N-glycans by MALDI-linear ion trap mass spectrometry
Automated measurement of permethylated serum N-glycans by MALDI-linear ion trap mass spectrometry
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DOI：
10.1016/j.carres.2009.06.010
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发表时间：
2009-08-17
影响因子：
3.1
通讯作者：
Lefeber, Dirk J.
中科院分区：
文献类型：
--
作者：
Guillard, Mailys;Gloerich, Jolein;Lefeber, Dirk J.
使用N-聚糖质谱进行临床诊断需要开发稳健的高通量分析方法。然而，聚糖的结构分配需要额外的信息，如MS 2片段化或外切糖苷酶二聚体。我们提出了一种设置，它结合了MALDI电离源与线性离子阱分析仪。由于晶体定位系统与MSn测序选项相结合，该仪器可以自动测量样品。通常用于聚糖分析的2，5-二羟基苯甲酸由于其非均匀结晶特性而不能产生所需的再现性。相反，α-氰基-4-羟基肉桂酸提供了均匀的结晶模式和测量的再现性。使用血清N-聚糖作为测试样品，我们通过优化仪器设置，专注于数据收集的自动化。通过MS 2分析确认聚糖结构。尽管样品处理仍需要优化，但该方法提供了一种使用MALDI线性离子阱仪器测量N-聚糖的可重复和高通量方法。(C)2009爱思唯尔有限公司版权所有。
The use of N-glycan mass spectrometry for clinical diagnostics requires the development of robust high-throughput profiling methods. Still, structural assignment of glycans requires additional information such as MS2 fragmentation or exoglycosidase digestions. We present a setting which combines a MALDI ionization source with a linear ion trap analyzer. This instrumentation allows automated measurement of samples thanks to the crystal positioning system, combined with MSn sequencing options. 2,5-Dihydroxybenzoic acid, commonly used for the analysis of glycans, failed to produce the required reproducibility due to its non-homogeneous crystallization properties. In contrast, alpha-cyano-4-hydroxycinnamic acid provided a homogeneous crystallization pattern and reproducibility of the measurements. Using serum N-glycans as a test sample, we focused on the automation of data collection by optimizing the instrument settings. Glycan structures were confirmed by MS2 analysis. Although sample processing still needs optimization, this method provides a reproducible and high-throughput approach for measurement of N-glycans using a MALDI-linear ion trap instrument. (C) 2009 Elsevier Ltd. All rights reserved.