Comparison of analytical methods for profiling N- and O-linked glycans from cultured cell lines: HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study
Author(s)
Ito, Hiromi
Kaji, Hiroyuki
Togayachi, Akira
Azadi, Parastoo
Ishihara, Mayumi
Geyer, Rudolf
Galuska, Christina
Geyer, Hildegard
Kakehi, Kazuaki
Kinoshita, Mitsuhiro
Karlsson, Niclas G
Jin, Chunsheng
Kato, Koichi
Yagi, Hirokazu
Kondo, Sachiko
Kawasaki, Nana
Hashii, Noritaka
Kolarich, Daniel
Stavenhagen, Kathrin
Packer, Nicolle H
Thaysen-Andersen, Morten
Nakano, Miyako
Taniguchi, Naoyuki
Kurimoto, Ayako
Wada, Yoshinao
Tajiri, Michiko
Yang, Pengyuan
Cao, Weiqian
Li, Hong
Rudd, Pauline M
Narimatsu, Hisashi
Year published
2016
Metadata
Show full item recordAbstract
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 ...
View more >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.
View less >
View more >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.
View less >
Journal Title
Glycoconjugate Journal
Volume
33
Issue
3
Subject
Biochemistry and cell biology
Biochemistry and cell biology not elsewhere classified
Medical microbiology
Neurosciences