Comparison of analytical methods for profiling N- and O-linked glycans from cultured cell lines

HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study

Hiromi Ito, Hiroyuki Kaji, Akira Togayachi, Parastoo Azadi, Mayumi Ishihara, Rudolf Geyer, Christina Galuska, Hildegard Geyer, Kazuaki Kakehi, Mitsuhiro Kinoshita, Niclas G. Karlsson, Chunsheng Jin, Koichi Kato, Hirokazu Yagi, Sachiko Kondo, Nana Kawasaki, Noritaka Hashii, Daniel Kolarich, Kathrin Stavenhagen, Nicolle H. Packer & 11 others Morten Thaysen-Andersen, Miyako Nakano, Naoyuki Taniguchi, Ayako Kurimoto, Yoshinao Wada, Michiko Tajiri, Pengyuan Yang, Weiqian Cao, Hong Li, Pauline M. Rudd*, Hisashi Narimatsu

*Corresponding author for this work

    Research output: Contribution to journalArticle

    16 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)405-415
    Number of pages11
    JournalGlycoconjugate Journal
    Volume33
    Issue number3
    DOIs
    Publication statusPublished - 1 Jun 2016

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