Comparison of methods for profiling O-glycosylation: Human Proteome Organisation Human Disease Glycomics/Proteome Initiative multi-institutional study of IgA1.
The Human Proteome Organisation Human Disease Glycomics/Proteome Initiative recently coordinated a multi-institutional study that evaluated methodologies that are widely used for defining the N-glycan content in glycoproteins. The study convincingly endorsed mass spectrometry as the technique of choice for glycomic profiling in the discovery phase of diagnostic research. The present study reports the extension of the Human Disease Glycomics/Proteome Initiative's activities to an assessment of the methodologies currently used for O-glycan analysis. Three samples of IgA1 isolated from the serum of patients with multiple myeloma were distributed to 15 laboratories worldwide for O-glycomics analysis. A variety of mass spectrometric and chromatographic procedures representative of current methodologies were used. Similar to the previous N-glycan study, the results convincingly confirmed the pre-eminent performance of MS for O-glycan profiling. Two general strategies were found to give the most reliable data, namely direct MS analysis of mixtures of permethylated reduced glycans in the positive ion mode and analysis of native reduced glycans in the negative ion mode using LC-MS approaches. In addition, mass spectrometric methodologies to analyze O-glycopeptides were also successful.
Wada
Yoshinao
Y
Osaka Medical Center and Research Institute for Maternal and Child Health, Izumi, Osaka, Japan. waday@mch.pref.osaka.jp
Dell
Anne
A
Haslam
Stuart M
SM
Tissot
Bérangère
B
Canis
Kévin
K
Azadi
Parastoo
P
Bäckström
Malin
M
Costello
Catherine E
CE
Hansson
Gunnar C
GC
Hiki
Yoshiyuki
Y
Ishihara
Mayumi
M
Ito
Hiromi
H
Kakehi
Kazuaki
K
Karlsson
Niclas
N
Hayes
Catherine E
CE
Kato
Koichi
K
Kawasaki
Nana
N
Khoo
Kay-Hooi
KH
Kobayashi
Kunihiko
K
Kolarich
Daniel
D
Kondo
Akihiro
A
Lebrilla
Carlito
C
Nakano
Miyako
M
Narimatsu
Hisashi
H
Novak
Jan
J
Novotny
Milos V
MV
Ohno
Erina
E
Packer
Nicolle H
NH
Palaima
Elizabeth
E
Renfrow
Matthew B
MB
Tajiri
Michiko
M
Thomsson
Kristina A
KA
Yagi
Hirokazu
H
Yu
Shin-Yi
SY
Taniguchi
Naoyuki
N
D000465:Algorithms; D002240:Carbohydrate Sequence; D004194:Disease / Q000209:etiology; D054794:Glycomics / Q000379:methods* / Q000458:organization & administration / Q000592:standards; D006023:Glycoproteins / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D055442:Metabolome; D008954:Models, Biological; D011134:Polysaccharides / Q000737:chemistry; D020543:Proteome / Q000032:analysis / Q000378:metabolism; D040901:Proteomics / Q000379:methods* / Q000458:organization & administration* / Q000592:standards; D012958:Societies, Scientific / Q000458:organization & administration
D006023:Glycoproteins; D007070:Immunoglobulin A; D011134:Polysaccharides; D020543:Proteome