Comparative performance of four methods for high-throughput glycosylation analysis of immunoglobulin G in genetic and epidemiological research.
The biological and clinical relevance of glycosylation is becoming increasingly recognized, leading to a growing interest in large-scale clinical and population-based studies. In the past few years, several methods for high-throughput analysis of glycans have been developed, but thorough validation and standardization of these methods is required before significant resources are invested in large-scale studies. In this study, we compared liquid chromatography, capillary gel electrophoresis, and two MS methods for quantitative profiling of N-glycosylation of IgG in the same data set of 1201 individuals. To evaluate the accuracy of the four methods we then performed analysis of association with genetic polymorphisms and age. Chromatographic methods with either fluorescent or MS-detection yielded slightly stronger associations than MS-only and multiplexed capillary gel electrophoresis, but at the expense of lower levels of throughput. Advantages and disadvantages of each method were identified, which should inform the selection of the most appropriate method in future studies.
From the ‡MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK;
Jennifer E
JE
Huffman
§Genos Glycoscience Laboratory, Zagreb, Croatia;
Maja
M
Pučić-Baković
§Genos Glycoscience Laboratory, Zagreb, Croatia;
Lucija
L
Klarić
¶Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; ‖glyXera GmbH, Magdeburg, Germany;
René
R
Hennig
**Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands;
Maurice H J
MH
Selman
§Genos Glycoscience Laboratory, Zagreb, Croatia;
Frano
F
Vučković
§Genos Glycoscience Laboratory, Zagreb, Croatia;
Mislav
M
Novokmet
§Genos Glycoscience Laboratory, Zagreb, Croatia;
Jasminka
J
Krištić
‖glyXera GmbH, Magdeburg, Germany;
Matthias
M
Borowiak
¶Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; ‖glyXera GmbH, Magdeburg, Germany;
Thilo
T
Muth
‡‡Faculty of Medicine, University of Split, Split, Croatia;
Ozren
O
Polašek
§§University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia;
Genadij
G
Razdorov
§§University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia;
Olga
O
Gornik
**Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands;
Rosina
R
Plomp
¶¶Centre for Population Health Sciences, School of Medicine and Veterinary Medicine, University of Edinburgh, Edinburgh, UK;
Evropi
E
Theodoratou
From the ‡MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK;
Alan F
AF
Wright
¶¶Centre for Population Health Sciences, School of Medicine and Veterinary Medicine, University of Edinburgh, Edinburgh, UK;
Igor
I
Rudan
From the ‡MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK;
Caroline
C
Hayward
¶¶Centre for Population Health Sciences, School of Medicine and Veterinary Medicine, University of Edinburgh, Edinburgh, UK;
Harry
H
Campbell
**Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands;
André M
AM
Deelder
¶Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; ‖‖Otto-von-Guericke University, Chair of Bioprocess Engineering, Magdeburg, Germany;
Udo
U
Reichl
Institute of Cytology and Genetics SB RAS, Novosibirsk, Russia; "Yurii Aulchenko" consulting, Groningen, The Netherlands;
Yurii S
YS
Aulchenko
¶Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; ‖glyXera GmbH, Magdeburg, Germany;
Erdmann
E
Rapp
**Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands; Division of BioAnalytical Chemistry, VU University Amsterdam, Amsterdam, The Netherlands.
Manfred
M
Wuhrer
§Genos Glycoscience Laboratory, Zagreb, Croatia; §§University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia; glauc@pharma.hr.
Gordan
G
Lauc
D000328:Adult; D002853:Chromatography, Liquid; D019075:Electrophoresis, Capillary; D006031:Glycosylation; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000235:genetics*; D013058:Mass Spectrometry / Q000379:methods*; D011110:Polymorphism, Genetic; D011134:Polysaccharides / Q000235:genetics* / Q000302:isolation & purification
D007074:Immunoglobulin G; D011134:Polysaccharides