title: Comparison of analytical methods for profiling N- and O-linked glycans from cultured cell lines : HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study.
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.
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8568, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ito
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8568, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kaji
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8568, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Togayachi
authors.affiliation: Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens 30602, USA.
authors.forename: Parastoo
authors.initials: P
authors.lastname: Azadi
authors.affiliation: Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens 30602, USA.
authors.forename: Mayumi
authors.initials: M
authors.lastname: Ishihara
authors.affiliation: Institute of Biochemistry, Faculty of Medicine, University of Giessen, Giessen D-35392, Germany.
authors.forename: Rudolf
authors.initials: R
authors.lastname: Geyer
authors.affiliation: Institute of Biochemistry, Faculty of Medicine, University of Giessen, Giessen D-35392, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Galuska
authors.affiliation: Institute of Biochemistry, Faculty of Medicine, University of Giessen, Giessen D-35392, Germany.
authors.forename: Hildegard
authors.initials: H
authors.lastname: Geyer
authors.affiliation: Department of Pharmaceutical Sciences, Faculty of Pharmacy, Kinki University, Osaka 577-8502, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Kakehi
authors.affiliation: Department of Pharmaceutical Sciences, Faculty of Pharmacy, Kinki University, Osaka 577-8502, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Kinoshita
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg 40530, Sweden.
authors.forename: Niclas G
authors.initials: NG
authors.lastname: Karlsson
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg 40530, Sweden.
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Jin
authors.affiliation: Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467-8603, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Kato
authors.affiliation: Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467-8603, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Yagi
authors.affiliation: Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467-8603, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kondo
authors.affiliation: Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Tokyo 158-8501, Japan.
authors.forename: Nana
authors.initials: N
authors.lastname: Kawasaki
authors.affiliation: Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Tokyo 158-8501, Japan.
authors.forename: Noritaka
authors.initials: N
authors.lastname: Hashii
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam D-14424, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kolarich
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam D-14424, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Stavenhagen
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney 2109, Australia.
authors.forename: Nicolle H
authors.initials: NH
authors.lastname: Packer
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney 2109, Australia.
authors.forename: Morten
authors.initials: M
authors.lastname: Thaysen-Andersen
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney 2109, Australia.
authors.forename: Miyako
authors.initials: M
authors.lastname: Nakano
authors.affiliation: Disease Glycomics Team, RIKEN, Saitama 351-0198, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
authors.affiliation: Disease Glycomics Team, RIKEN, Saitama 351-0198, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Kurimoto
authors.affiliation: Osaka Medical Center and Research Institute for Maternal and Child Health, Osaka 594-1101, Japan.
authors.forename: Yoshinao
authors.initials: Y
authors.lastname: Wada
authors.affiliation: Osaka Medical Center and Research Institute for Maternal and Child Health, Osaka 594-1101, Japan.
authors.forename: Michiko
authors.initials: M
authors.lastname: Tajiri
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Pengyuan
authors.initials: P
authors.lastname: Yang
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Weiqian
authors.initials: W
authors.lastname: Cao
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.affiliation: National Institute for Bioprocessing Research and Training (NIBRT), Dublin, Ireland.
authors.forename: Pauline M
authors.initials: PM
authors.lastname: Rudd
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8568, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Narimatsu
mesh_terms: D015415:Biomarkers / Q000737:chemistry; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D054794:Glycomics / Q000379:methods* / Q000592:standards; D006023:Glycoproteins / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000592:standards; D011134:Polysaccharides / Q000737:chemistry*; D040901:Proteomics / Q000379:methods / Q000592:standards; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; D006023:Glycoproteins; D011134:Polysaccharides
keywords: Glycoproteomics; Human disease glycomics/proteome initiative (HGPI); Human proteome organization (HUPO)