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首页> 外文期刊>Analytica chimica acta >Relative quantitation of neutral and sialylated N-glycans using stable isotopic labeled d0/d5-benzoyl chloride by MALDI-MS
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Relative quantitation of neutral and sialylated N-glycans using stable isotopic labeled d0/d5-benzoyl chloride by MALDI-MS

机译：使用MALDI-MS使用稳定同位素标记的D0 / D5-苯甲酰氯的中性和唾液酸唾液酸N-聚糖的相对定量

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摘要

Quantitative analysis of glycans is an emerging field in glycomic research. Herein we present a rapid and effective dual-labeling strategy, in the combination of isotopic derivatization of N-glycosylamine-based glycans by d0/d5-benzoyl chloride and methylamidation of sialic acids, to relatively quantify both neutral and sialylated N-glycans simultaneously by MALDI-MS. The derivatization efficiencies were increased by microwave-accelerated deglycosylation which not only largely reduce the time of glycoprotein deglycosylation but also inhibit the hydrolysis of intermediate glycosylamines produced by PNGase F digestion. Three model glycoproteins, including RNase B, bovine fetuin and IgG from human serum, were applied to validate this technique. Results showed that the glycans from microgram level of glycoprotein can be successfully quantified with high reproducibility and the whole time of analytical procedure was shortened to 4 h. Furthermore, this proposed method was applied for the comparative analysis of N-glycans from serum of healthy donors and multiple myeloma patients. It was found that five N-glycans may be as the potential biomarkers for rapid detection of early multiple myeloma, indicating the feasibility of this strategy in monitoring subtle quantitative differences of serum glycomics. (c) 2017 Elsevier B.V. All rights reserved.
机译：聚糖的定量分析是glycomic研究中的一个新兴领域。在此我们提出一个快速有效的双标记策略，在N基葡基聚糖由D0 / D5 - 苯甲酰氯和唾液酸methylamidation的同位素衍生的结合，以相对同时通过量化中性和唾液酸化的N-聚糖MALDI-MS。衍生化的效率分别增加了微波加速的去糖基化这不仅大大降低糖蛋白去糖基化的时间，而且还抑制由PNG酶消化˚F产生的中间葡基胺的水解。三个模型糖蛋白，包括RNA酶B，牛胎球蛋白和IgG从人血清中，分别适用于验证该技术。结果表明，从糖蛋白的微克水平上的聚糖可以以高再现性和分析方法的整个时间缩短至4小时成功地定量。另外，涂敷了用于N-聚糖的来自健康供体和多发性骨髓瘤患者的血清中的比较分析此提出的方法。结果发现，5 N-聚糖可以是作为用于快速检测早期多发性骨髓瘤的潜在生物标志物，指示从该策略在监测血清糖组学的细微定量差异的可行性。 （c）2017 Elsevier B.v.保留所有权利。

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  • 来源
    《Analytica chimica acta》 |2018年第2018期|共12页
  • 作者

    Wang Chang; Wu Yike; Zhang Liang; Liu Bi-Feng; Lin Yawei; Liu Xin;

  • 作者单位

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol MOE Wuhan Natl Lab Optoel Dept Biomed Engn Key Lab Biomed Photon Hubei Bioinformat &

    Mol Imaging Key Lab Syst Biol Wuhan 430074 Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol MOE Wuhan Natl Lab Optoel Dept Biomed Engn Key Lab Biomed Photon Hubei Bioinformat &

    Mol Imaging Key Lab Syst Biol Wuhan 430074 Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol MOE Wuhan Natl Lab Optoel Dept Biomed Engn Key Lab Biomed Photon Hubei Bioinformat &

    Mol Imaging Key Lab Syst Biol Wuhan 430074 Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol MOE Wuhan Natl Lab Optoel Dept Biomed Engn Key Lab Biomed Photon Hubei Bioinformat &

    Mol Imaging Key Lab Syst Biol Wuhan 430074 Hubei Peoples R China;

    Wuhan Univ Technol Sch Chem Chem Engn &

    Life Sci Wuhan 430070 Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol MOE Wuhan Natl Lab Optoel Dept Biomed Engn Key Lab Biomed Photon Hubei Bioinformat &

    Mol Imaging Key Lab Syst Biol Wuhan 430074 Hubei Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
  • 关键词

    Rapid deglycosylation; d0/d5-benzoyl chloride; N-glycosylamine; Stable isotopic labeling; N-glycans; Biomarker;

    机译：快速脱糖基化;D0 / D5-苯甲酰氯;N-糖基胺;稳定同位素标记;N-聚糖;生物标志物;

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