Comparative Glycomic Mapping through Quantitative Permethylation and Stable-Isotope Labeling

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• 作者：Pilsoo Kang ; Yehia Mechref ; Zuzana Kyselova ; John A. Goetz ; Milos V. Novotny
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：August 15, 2007
• 年：2007
• 卷：79
• 期：16
• 页码：6064 - 6073
• 全文大小：952K
• 年卷期：v.79,no.16(August 15, 2007)
• ISSN：1520-6882

文摘

Comparative glycan quantification has thus far been achallenging task due to the lack of sensitive and reproducible analytical techniques. We introduce here a combination of quantitative permethylation and isotope labeling of glycans as an approach (C-GlycoMAP) allowingprecise comparison between different samples in a singleMALDI-MS analysis. Samples are either methylated ordeuteriomethylated prior to their mixing and mass spectrometric acquisitions. Comparative analyses are basedon the ratio of the two isotopically distinct forms of thesame glycan structure, thus allowing a direct absoluteevaluation of the intensities of the two forms originatingfrom two different biological samples (e.g., control anddiseased). The direct comparison between the two formseliminates a MALDI-MS low m/z bias commonly associated with this technique. These comparative analyses arehighly reliable when the intensity ratios of the two formslie between 0.125 and 6, an overall reproducibility betterthan 30% (RSD). The value of C-GlycoMAP is demonstrated here for N-glycans derived from human bloodserum collected from a healthy individual and a breastcancer patient as well as for O-glycans derived fromnormal and cancer cell extracts.