Simplified Quantitative Glycomics Using the Stable Isotope Label Girard's Reagent P by Electrospray Ionization Mass Spectrometry
Simplified Quantitative Glycomics Using the Stable Isotope Label Girard's Reagent P by Electrospray Ionization Mass Spectrometry
复制标题
通过电喷雾电离质谱法使用稳定同位素标记吉拉德试剂 P 简化定量糖组学
DOI：
10.1021/pr4010647
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发表时间：
2014
影响因子：
4.4
通讯作者：
Huang Linjuan
中科院分区：
文献类型：
--
作者：
Wang Chengjian;Wu Zhiyu;Yuan Jiangbei;Wang Bo;Zhang Ping;Zhang Ying;Wang Zhongfu;Huang Linjuan
快速，灵敏，简单的方法定量分析不同生物样品之间的聚糖表达的差异是必不可少的蛋白质糖基化模式（糖组学）的研究和疾病的聚糖生物标志物的搜索。基于稳定同位素标记结合质谱检测的聚糖相对定量代表了一种新兴且有前途的技术。然而，这种技术被破坏的同位素标记的聚糖的质谱的复杂性所造成的多个金属离子加合物信号的存在下，这导致在检测灵敏度的降低和增加的困难，在数据解释。本文报道了一种简化的定量糖组学策略，其特征在于用非氘代（d 0-）或氘代（d5-）吉拉德试剂P（GP）对还原性聚糖进行非还原性同位素标记，而不引入盐，并简化了中性聚糖的d 0-和d5-GP衍生物作为分子离子的质谱图，而不需要复杂的金属离子加合物，允许在不同聚糖样品之间进行快速和灵敏的定量比较。我们得到了优化的GP标记条件和良好的定量线性，重现性和数据的准确性的方法。通过对牛和猪免疫球蛋白G以及小鼠和大鼠血清中中性N-聚糖的比较定量分析，验证了该方法的良好适用性。此外，我们已经揭示了这种策略的潜力，作为GP衍生物的唾液酸化聚糖的高灵敏度分析，其中包括通过化学衍生化的唾液酸的羧基的中和。
Fast, sensitive, and simple methods for quantitative analysis of disparities in glycan expression between different biological samples are essential for studies of protein glycosylation patterns (glycomics) and the search for disease glycan biomarkers. Relative quantitation of glycans based on stable isotope labeling combined with mass spectrometric detection represents an emerging and promising technique. However, this technique is undermined by the complexity of mass spectra of isotope-labeled glycans caused by the presence of multiple metal ion adduct signals, which result in a decrease of detection sensitivity and an increase of difficulties in data interpretation. Herein we report a simplified quantitative glycomics strategy, which features nonreductive isotopic labeling of reducing glycans with either nondeuterated (d0-) or deuterated (d5-) Girard’s reagent P (GP) without salts introduced and simplified mass spectrometric profiles ofd0- andd5-GP derivatives of neutral glycans as molecular ions without complex metal ion adducts, allowing rapid and sensitive quantitative comparison between different glycan samples. We have obtained optimized GP-labeling conditions and good quantitation linearity, reproducibility, and accuracy of data by the method. Its excellent applicability was validated by comparatively quantitative analysis of the neutralN-glycans released from bovine and porcine immunoglobulin G as well as of those from mouse and rat sera. Additionally, we have revealed the potential of this strategy for the high-sensitivity analysis of sialylated glycans as GP derivatives, which involves neutralization of the carboxyl group of sialic acid by chemical derivatization.