Longitudinal monitoring of immunoglobulin A glycosylation during pregnancy by simultaneous MALDI-FTICR-MS analysis of N- and O-glycopeptides.
Immunoglobulin A (IgA) is a glycoprotein of which altered glycosylation has been associated with several pathologies. Conventional methods for IgA N- and O-glycosylation analysis are tedious, thus limiting such analyses to small sample sizes. Here we present a high-throughput strategy for the simultaneous analysis of serum-derived IgA1 N- and O-glycopeptides using matrix-assisted laser/desorption ionisation Fourier transform ion cyclotron resonance (MALDI-FTICR) mass spectrometry (MS). Six non-fucosylated diantennary complex type glycoforms were detected on the Asn144-containing glycopeptide. Thirteen distinct glycoforms were identified for the Asn340-containing tailpiece glycopeptide, mainly of the diantennary complex type, and low amounts of triantennary glycoforms. Simultaneously with these N-glycopeptides, 53 compositional glycoforms of the hinge region O-glycopeptide were profiled in a single high resolution MALDI-FTICR spectrum. Since many pregnancy associated changes have been recognized for immunoglobulin G, we sought to demonstrate the clinical applicability of this method in a cohort of 29 pregnant women, from whom samples were collected at three time points during pregnancy and three time points after delivery. Pregnancy associated changes of N-glycan bisection were different for IgA1 as compared to IgG-Fc described earlier. We foresee further applications of the developed method for larger patient cohorts to study IgA N- and O-glycosylation changes in pathologies.
Department of Rheumatology, Erasmus University Medical Center, Rotterdam, The Netherlands.|Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Albert
A
Bondt
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Simone
S
Nicolardi
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Bas C
BC
Jansen
Division of BioAnalytical Chemistry, VU University Amsterdam, The Netherlands.
Kathrin
K
Stavenhagen
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Dennis
D
Blank
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Guinevere S M
GS
Kammeijer
Ludger, Culham Science Centre, Oxfordshire OX14 3EB, UK.
Radoslaw P
RP
Kozak
Ludger, Culham Science Centre, Oxfordshire OX14 3EB, UK.
Daryl L
DL
Fernandes
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Paul J
PJ
Hensbergen
Department of Rheumatology, Erasmus University Medical Center, Rotterdam, The Netherlands.
Johanna M W
JM
Hazes
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Yuri E M
YE
van der Burgt
Department of Rheumatology, Erasmus University Medical Center, Rotterdam, The Netherlands.
Radboud J E M
RJ
Dolhain
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.|Division of BioAnalytical Chemistry, VU University Amsterdam, The Netherlands.
Manfred
M
Wuhrer
D005260:Female; D006031:Glycosylation; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood / Q000737:chemistry*; D007155:Immunologic Factors / Q000097:blood / Q000737:chemistry*; D008137:Longitudinal Studies; D011134:Polysaccharides / Q000032:analysis*; D011247:Pregnancy; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017550:Spectroscopy, Fourier Transform Infrared
D007070:Immunoglobulin A; D007155:Immunologic Factors; D011134:Polysaccharides