Comparison of analytical methods for profiling N- and O-linked glycans from cultured cell lines : HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study.
Comparison of analytical methods for profiling N- and O-linked glycans from cultured cell lines : HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study.
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比较分析培养细胞系的N和O连接聚糖的分析方法：HUPO人类疾病糖/蛋白质组倡议多机构研究。
DOI：
10.1007/s10719-015-9625-3
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发表时间：
2016-06
影响因子：
3
通讯作者：
Narimatsu H
中科院分区：
文献类型：
--
作者：
Ito H;Kaji H;Togayachi A;Azadi P;Ishihara M;Geyer R;Galuska C;Geyer H;Kakehi K;Kinoshita M;Karlsson NG;Jin C;Kato K;Yagi H;Kondo S;Kawasaki N;Hashii N;Kolarich D;Stavenhagen K;Packer NH;Thaysen-Andersen M;Nakano M;Taniguchi N;Kurimoto A;Wada Y;Tajiri M;Yang P;Cao W;Li H;Rudd PM;Narimatsu H
人类疾病糖组/蛋白质组倡议(HGPI)是人类蛋白质组组织(HUPO)的一项活动，由国际研究机构的领先研究人员支持，旨在开发与疾病相关的糖组/糖蛋白质组分析技术。自2004年以来，该倡议进行了三项试点研究。前两项是对纯化的转铁蛋白和免疫球蛋白-G进行N-和O-葡聚糖分析，并评估当时采用的最合适的分析方法。本文描述了第三项研究，该研究比较了不同的方法来定量粗制生物样品中附着在蛋白质上的N-和O-连接的葡聚糖。对细胞颗粒的初步分析产生了差异很大的葡聚糖谱，这可能是前处理样品制备方法不同的结果。然而，在随后由一家实验室以特定方法制备的细胞裂解物组分的分析中，数据的重复性并未得到显著改善。这项研究表明，用任何单一技术都很难对原油样品中的糖苷进行完整的分析，以及从前处理到数据解释的分析过程严格优化的重要性。这表明，在这一快速发展的领域中采用最新技术的另一项合作研究将有助于实现对复杂细胞样本中糖蛋白进行大规模分析的要求。
The Human Disease Glycomics/Proteome Initiative (HGPI) is an activity in the Human Proteome Organization (HUPO) supported by leading researchers from international institutes and aims at development of disease-related glycomics/glycoproteomics analysis techniques. Since 2004, the initiative has conducted three pilot studies. The first two were N- and O-glycan analyses of purified transferrin and immunoglobulin-G and assessed the most appropriate analytical approach employed at the time. This paper describes the third study, which was conducted to compare different approaches for quantitation of N- and O-linked glycans attached to proteins in crude biological samples. The preliminary analysis on cell pellets resulted in wildly varied glycan profiles, which was probably the consequence of variations in the pre-processing sample preparation methodologies. However, the reproducibility of the data was not improved dramatically in the subsequent analysis on cell lysate fractions prepared in a specified method by one lab. The study demonstrated the difficulty of carrying out a complete analysis of the glycome in crude samples by any single technology and the importance of rigorous optimization of the course of analysis from preprocessing to data interpretation. It suggests that another collaborative study employing the latest technologies in this rapidly evolving field will help to realize the requirements of carrying out the large-scale analysis of glycoproteins in complex cell samples.