Comparison of Methods for Profiling O-Glycosylation

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.

[1]  J. Mestecky,et al.  Receptor-mediated binding and uptake of immunoglobulin A by human liver. , 1988, Gastroenterology.

[2]  M. J. Chalmers,et al.  Analysis of O-glycan heterogeneity in IgA1 myeloma proteins by Fourier transform ion cyclotron resonance mass spectrometry: implications for IgA nephropathy , 2007, Analytical and bioanalytical chemistry.

[3]  Yusuke Suzuki,et al.  Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity. , 2009, The Journal of clinical investigation.

[4]  Mark C. Field,et al.  O-linked oligosaccharides from human serum immunoglobulin A1. , 1989, Biochemical Society transactions.

[5]  J. Marth,et al.  Critical functions of N-glycans in L-selectin-mediated lymphocyte homing and recruitment , 2007, Nature Immunology.

[6]  H. Cooper,et al.  Determination of Aberrant O-Glycosylation in the IgA1 Hinge Region by Electron Capture Dissociation Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry* , 2005, Journal of Biological Chemistry.

[7]  Richard D Cummings,et al.  Altered O-glycosylation and sulfation of airway mucins associated with cystic fibrosis. , 2005, Glycobiology.

[8]  Kunihiko Kobayashi,et al.  J‐chain determinants in polymeric immunoglobulins , 1973, European journal of immunology.

[9]  M. Tajiri,et al.  Hydrophilic affinity isolation and MALDI multiple-stage tandem mass spectrometry of glycopeptides for glycoproteomics. , 2004, Analytical chemistry.

[10]  G. Carlone,et al.  Evaluation of monoclonal antibodies with specificity for human IgA, IgA subclasses and allotypes and secretory component. Results of an IUIS/WHO collaborative study. , 1996, Journal of immunological methods.

[11]  R. Zubarev,et al.  Localization of O-glycosylation sites in peptides by electron capture dissociation in a Fourier transform mass spectrometer. , 1999, Analytical chemistry.

[12]  J. Mestecky.  Immunobiology of IgA. , 1988, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[13]  B. Hendry,et al.  Human serum IgA1 is substituted with up to six O-glycans as shown by matrix assisted laser desorption ionisation time-of-flight mass spectrometry. , 2004, Carbohydrate research.

[14]  J. Peter-Katalinic,et al.  Electron Capture Dissociation of O-Glycosylated Peptides: Radical Site-Induced Fragmentation of Glycosidic Bonds , 2005, European journal of mass spectrometry.

[15]  R. Dwek,et al.  The Glycosylation and Structure of Human Serum IgA1, Fab, and Fc Regions and the Role of N-Glycosylation on Fcα Receptor Interactions* , 1998, The Journal of Biological Chemistry.

[16]  Naoyuki Taniguchi,et al.  Comparison of the methods for profiling glycoprotein glycans--HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study. , 2007, Glycobiology.

[17]  S. Kornfeld,et al.  Structure of the carbohydrate units of IgA1 immunoglobulin. II. Structure of the O-glycosidically linked oligosaccharide units. , 1974, The Journal of biological chemistry.

[18]  G. Alarcón,et al.  IgA1-secreting cell lines from patients with IgA nephropathy produce aberrantly glycosylated IgA1. , 2008, The Journal of clinical investigation.

[19]  B. Julian,et al.  Patients with IgA nephropathy have increased serum galactose-deficient IgA1 levels. , 2007, Kidney international.

[20]  Stuart,et al.  Structure of the Carbohydrate Units of IgA, Immunoglobulin , 2003 .

[21]  J. Novak,et al.  Heterogeneity of O-glycosylation in the hinge region of human IgA1. , 2000, Molecular immunology.