Stable-isotope labeled hydrophobic hydrazide reagents for the relative quantification of N-linked glycans by electrospray ionization mass spectrometry.
Stable-isotope labeled hydrophobic hydrazide reagents for the relative quantification of N-linked glycans by electrospray ionization mass spectrometry.
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DOI：
10.1021/ac201376q
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发表时间：
2011-09-01
影响因子：
7.4
通讯作者：
Muddiman, David C.
中科院分区：
文献类型：
--
作者：
Walker, S. Hunter;Budhathoki-Uprety, Januka;Novak, Bruce M.;Muddiman, David C.
本研究介绍了稳定同位素标记的疏水性酰肼试剂的开发，用于 N-连接聚糖的相对定量。 P2GPN“轻”(12C) 和“重”(13C6) 对用于差异标记两个 N 连接聚糖样品。将样品按 1:1 组合，使用 HILIC 进行分离，然后使用质谱进行质量区分和定量。这些试剂有几个优点：1) 赋予聚糖疏水性，从而提高电喷雾电离效率和 MS 检测； 2) “轻”和“重”试剂的色谱、MS 和 MS/MS 性能无法区分，无法进行相对定量； 3) 由于样品制备后将两个样品混合在一起，分析变异性显着降低。获得这些分析优势仅需要约 4 小时的样品制备时间。结果表明，这些试剂能够量化简单混合物中糖基化的变化，并且合并血浆样品中试剂的分析变异性小于±30%。此外，内标的加入可以解释两个样品之间系统误差的差异，因为样品是平行处理的，直到衍生化后才混合。
This study presents the development of stable-isotope labeled hydrophobic, hydrazide reagents for the relative quantification of N-linked glycans. The P2GPN ‘light’ (12C) and ‘heavy’ (13C6) pair are used to differentially label two N-linked glycan samples. The samples are combined 1:1, separated using HILIC, and then mass differentiated and quantified using mass spectrometry. These reagents have several benefits: 1) impart hydrophobic character to the glycans affording an increase in electrospray ionization efficiency and MS detection; 2) indistinguishable chromatographic, MS, and MS/MS performance of the ‘light’ and ‘heavy’ reagents affording relative quantification; and 3) analytical variability is significantly reduced due to the two samples being mixed together after sample preparation. Obtaining these analytical benefits only requires ~4 hours of sample preparation time. It is shown that these reagents are capable of quantifying changes in glycosylation in simple mixtures, and the analytical variability of the reagents in pooled plasma samples is shown to be less than ±30%. Additionally, the incorporation of an internal standard allows one to account for the difference in systematic error between the two samples due to the samples being processed in parallel and not mixed until after derivatization.
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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：
--
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：
7
作者：
Kirmiz, Crystal;Li, Bensheng;Miyamoto, Suzanne
通讯作者：
Miyamoto, Suzanne
影响因子：
4.3
作者：
Alvarez-Manilla, Gerardo;Warren, Nicole Lynn;Orlando, Ron
通讯作者：
Orlando, Ron
影响因子：
4
作者：
Casey, RC;Oegema, TR;Skubitz, APN
通讯作者：
Skubitz, APN