Development of a robust and high throughput method for profiling N-linked glycans derived from plasma glycoproteins by NanoLC-FTICR mass spectrometry.
Development of a robust and high throughput method for profiling N-linked glycans derived from plasma glycoproteins by NanoLC-FTICR mass spectrometry.
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DOI：
10.1021/pr9002323
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发表时间：
2009-07
影响因子：
4.4
通讯作者：
Muddiman DC
中科院分区：
文献类型：
--
作者：
Bereman MS;Young DD;Deiters A;Muddiman DC
最近的研究继续强调糖基化在包括癌症在内的各种疾病中的重要性。在这项工作中，我们提出了一个一步一步的协议，描述了一种方法的N-连接的聚糖分析血浆糖蛋白的纳米流液相色谱傅立叶变换离子回旋共振质谱（FT-ICR-MS）。一个大的实验空间进行了初步探索，并在此描述。将三种内标物加标至样品中，并提供不同实验条件下血浆聚糖丰度的归一化。探索孵育方法、时间和NP 40去污剂对聚糖丰度的影响。结果发现，18小时的孵育无洗涤剂导致最大的离子丰度;然而，数据可以在不到一天的时间内从原始血浆样品利用微波辐射或更短的孵育期。当在初始处理步骤前加入内标物时，三种不同聚糖的样品间精密度小于5.5%（RSD）。FT-ICR质谱仪提供的高质量测量准确度（<3 ppm）为几种聚糖种类提供了可靠的鉴定。
Recent investigations continue to emphasize the importance of glycosylation in various diseases including cancer. In this work, we present a step by step protocol describing a method for N-linked glycan profiling of plasma glycoproteins by nano-flow liquid chromatography Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). A large experimental space was initially explored and is described herein. Three internal standards were spiked into the sample and provided normalization of plasma glycan abundance across different experimental conditions. Incubation methods, times and the effect of NP40 detergent on glycan abundance were explored. It was found that an 18-hour incubation with no detergent lead to the greatest ion abundance; however, data could be obtained in less than one day from raw plasma samples utilizing microwave irradiation or shorter incubation periods. The inter-sample precision of three different glycans was less than 5.5% (RSD) when the internal standards were added prior to the initial processing step. The high mass measurement accuracy (<3 ppm) afforded by the FT-ICR mass spectrometer provided confident identifications of several glycan species.
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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
DOI：
10.1016/j.jchromb.2009.02.040
发表时间：
2009-04-01
影响因子：
3
作者：
Andrews, Genna L.;Shuford, Christopher M.;Burnett, John C., Jr.;Hawkridge, Adam M.;Muddiman, David C.
通讯作者：
Muddiman, David C.
影响因子：
7
作者：
Kirmiz, Crystal;Li, Bensheng;Miyamoto, Suzanne
通讯作者：
Miyamoto, Suzanne
影响因子：
4
作者：
Casey, RC;Oegema, TR;Skubitz, APN
通讯作者：
Skubitz, APN
影响因子：
7.4
作者：
Ruhaak, L. Renee;Huhn, Carolin;Wuhrer, Manfred
通讯作者：
Wuhrer, Manfred