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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >Development of a filter-aided extraction method coupled with glycosylamine labeling to simplify and enhance high performance liquid chromatography-based N-glycan analysis
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Development of a filter-aided extraction method coupled with glycosylamine labeling to simplify and enhance high performance liquid chromatography-based N-glycan analysis

机译：滤波辅助提取方法的研制耦合糖基胺标记，以简化和增强高效液相色谱的基于N-Glycan分析

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摘要

Efficient sample pretreatment of N-glycans from glycoproteins is essential but challenging due to the limitations of existing tedious and laborious methods in N-glycomics. This study aimed to establish a filter-aided extraction method coupled with glycosylamine AQC labeling for a simple and rapid direct HPLC-FLD-based analysis of N-glycans. The developed method was demonstrated to be simpler and more sensitive compared to previous HILIC SPE purification method coupled with glycosylamine labeling. It has been validated with wild-type N-glycans from human transferrin and RNase B and then was successfully applied to investigate N-glycan profiles of the transferrin in human serum and a monoclonal antibody (mAb). Results showed good applicability of the method for complex samples. Additionally, this method is compatible with the replicate determination of N-glycan samples to assess the high-throughput analysis of glycan variability in mAb sample. (C) 2019 Elsevier B.V. All rights reserved.
机译：来自糖蛋白的N-聚糖的有效样品预处理是必不可少的，而是挑战，因为存在于N-Glycooces中存在的繁琐和富费的方法。 本研究旨在建立一种耦合的含有糖基胺AQC标记的滤波辅助提取方法，用于基于N-聚糖的简单和快速直接的HPLC-FLD分析。 与先前的HILIC SPE纯化方法与糖基胺标记相结合，表明开发方法更简单和更敏感。 它已被来自人转移素和RNase B的野生型N-聚糖验证，然后成功地应用于研究人血清中转移素的N-聚糖型材和单克隆抗体（MAb）。 结果表明，复杂样品的方法良好适用。 另外，该方法与重复测定N-Glycan样品以评估MAB样品中的聚糖变异性的高通量分析。 （c）2019 Elsevier B.v.保留所有权利。

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  • 作者

    Wu Yike; Sha Qiuyue; Wang Chang; Liu Bi-Feng; Wang Song; Liu Xin;

  • 作者单位

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol Dept Biomed Engn Wuhan Natl Lab Optoelect Syst Biol Theme Britton Chance Ctr Biomed Photon Wuhan 430074 Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol Dept Biomed Engn Wuhan Natl Lab Optoelect Syst Biol Theme Britton Chance Ctr Biomed Photon Wuhan 430074 Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol Dept Biomed Engn Wuhan Natl Lab Optoelect Syst Biol Theme Britton Chance Ctr Biomed Photon Wuhan 430074 Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol Dept Biomed Engn Wuhan Natl Lab Optoelect Syst Biol Theme Britton Chance Ctr Biomed Photon Wuhan 430074 Hubei Peoples R China;

    Wuhan Sports Univ Exercise Immunol Ctr Wuhan Hubei Peoples R China;

    Huazhong Univ Sci &

    Technol Coll Life Sci &

    Technol Dept Biomed Engn Wuhan Natl Lab Optoelect Syst Biol Theme Britton Chance Ctr Biomed Photon 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

    N-glycans; Glycosylamine labeling; Filter-aided extraction; HILIC-HPLC/FLD;

    机译：N-聚糖;糖基胺标记;滤波辅助提取;HILIC-HPLC / FLD;

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