Sensitive carbohydrate detection using surface enhanced Raman tagging.
Sensitive carbohydrate detection using surface enhanced Raman tagging.
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DOI：
10.1021/ac102284x
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发表时间：
2010-12-15
影响因子：
7.4
通讯作者：
Zhang, Dongmao
中科院分区：
文献类型：
--
作者：
Vangala, Karthikeshwar;Yanney, Michael;Hsiao, Cheng-Te;Wu, Wells W.;Shen, Rong-Fong;Zou, Sige;Sygula, Andrzej;Zhang, Dongmao
糖基化是蛋白质翻译后修饰中最重要的修饰之一，因此糖基化分析在生物和生物医学研究中是一个日益重要的领域。通过设计和应用罗丹明B衍生物作为表面增强拉曼光谱(SERS)标记，我们发展了一种检测碳水化合物的新技术。通过还原胺化反应，成功地将罗丹明标记(RT)连接到三种模型碳水化合物(葡萄糖、乳糖和葡萄糖醛酸)上。用632 nm HeNe激光获得的SERS检测下限为：所有RT-碳水化合物的浓度为~1 nm，总样品消耗量为~10fmol。SERS方法的动态范围约为4个数量级，范围从1 nm到5微米。使用同位素取代的SERS内部参照物的比率SERS定量方法还允许分别用RT标记的碳水化合物和用氘/氢取代的RT标记的碳水化合物的比较定量。除了增强标记碳水化合物的SERS检测外，罗丹明标记还促进了碳水化合物的荧光和质谱学检测。目前RT-碳水化合物的荧光灵敏度为~3 nm，而用线性离子陷阱电喷雾电离(ESI-MS)仪测得的MS灵敏度约为1fmol。讨论了利用这种SERS标记策略的高SERS、荧光和MS敏感性的潜在应用，用于实际的糖类分析，其中碳水化合物可以用荧光和SERS技术定量，然后用ESI-MS技术鉴定。
Glycomic analysis is an increasingly important field in biological and biomedical research as glycosylation is one of the most important protein post-translational modifications. We have developed a new technique to detect carbohydrates using surface enhanced Raman spectroscopy (SERS) by designing and applying a Rhodamine B derivative as the SERS tag. Using a reductive amination reaction, the Rhodamine-based tag (RT) was successfully conjugated to three model carbohydrates (glucose, lactose and glucuronic acid). SERS detection limits obtained with 632 nm HeNe laser were ~1 nM in concentration for all the RT-carbohydrate conjugates and ~10 fmol in total sample consumption. The dynamic range of the SERS method is about 4 orders of magnitude, spanning from 1 nM to 5 µM. Ratiometric SERS quantification using isotope-substituted SERS internal references also allows comparative quantifications of carbohydrates labeled with RT and deuterium/hydrogen substituted RT tags, respectively. In addition to enhancing the SERS detection of the tagged carbohydrates, the Rhodamine tagging facilitates fluorescence and mass spectrometric detection of carbohydrates. Current fluorescence sensitivity of RT-carbohydrates is ~ 3 nM in concentration while the mass spectrometry (MS) sensitivity is about 1 fmol that was achieved with linear ion trap electrospray ionization (ESI)-MS instrument. Potential applications that take advantage of the high SERS, fluorescence and MS sensitivity of this SERS tagging strategy are discussed for practical glycomic analysis where carbohydrates may be quantified with a fluorescence and SERS technique, and then identified with ESI-MS techniques.
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影响因子：
7.4
作者：
Mrozek, MF;Weaver, MJ
通讯作者：
Weaver, MJ
影响因子：
7.4
作者：
Goetz, John A.;Novotny, Milos V.;Mechref, Yehia
通讯作者：
Mechref, Yehia
影响因子：
4.3
作者：
Dalpathado, DS;Jiang, H;Desaire, H
通讯作者：
Desaire, H
葡萄糖醛酸化，吡咯里西啶生物碱的新代谢途径。
影响因子：
4.1
作者：
He, Yu-Qi;Yang, Li;Wang, Zheng-Tao
通讯作者：
Wang, Zheng-Tao
影响因子：
3.8
作者：
Char, Tom S.;Wilson, John X.;O'Brien, Peter J.
通讯作者：
O'Brien, Peter J.