30 results on '"Bas C. Jansen"'
Search Results
2. Large-Scale Analysis of Apolipoprotein CIII Glycosylation by Ultrahigh Resolution Mass Spectrometry
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Daniel Demus, Annemieke Naber, Viktoria Dotz, Bas C. Jansen, Marco R. Bladergroen, Jan Nouta, Eric J. G. Sijbrands, Mandy Van Hoek, Simone Nicolardi, and Manfred Wuhrer
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high-throughput ,mass spectrometry ,apolipoprotein-CIII ,glycosylation ,oxidation ,Chemistry ,QD1-999 - Abstract
Apolipoprotein-CIII (apo-CIII) is a glycoprotein involved in lipid metabolism and its levels are associated with cardiovascular disease risk. Apo-CIII sialylation is associated with improved plasma triglyceride levels and its glycosylation may have an effect on the clearance of triglyceride-rich lipoproteins by directing these particles to different metabolic pathways. Large-scale sample cohort studies are required to fully elucidate the role of apo-CIII glycosylation in lipid metabolism and associated cardiovascular disease. In this study, we revisited a high-throughput workflow for the analysis of intact apo-CIII by ultrahigh-resolution MALDI FT-ICR MS. The workflow includes a chemical oxidation step to reduce methionine oxidation heterogeneity and spectrum complexity. Sinapinic acid matrix was used to minimize the loss of sialic acids upon MALDI. MassyTools software was used to standardize and automate MS data processing and quality control. This method was applied on 771 plasma samples from individuals without diabetes allowing for an evaluation of the expression levels of apo-CIII glycoforms against a panel of lipid biomarkers demonstrating the validity of the method. Our study supports the hypothesis that triglyceride clearance may be regulated, or at least strongly influenced by apo-CIII sialylation. Interestingly, the association of apo-CIII glycoforms with triglyceride levels was found to be largely independent of body mass index. Due to its precision and throughput, the new workflow will allow studying the role of apo-CIII in the regulation of lipid metabolism in various disease settings.
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- 2021
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3. IgA N- and O-glycosylation profiling reveals no association with the pregnancy-related improvement in rheumatoid arthritis
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Albert Bondt, Simone Nicolardi, Bas C. Jansen, T. Martijn Kuijper, Johanna M. W. Hazes, Yuri E. M. van der Burgt, Manfred Wuhrer, and Radboud J. E. M. Dolhain
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Rheumatoid arthritis ,Pregnancy ,Immunoglobulin A ,Glycosylation ,Diseases of the musculoskeletal system ,RC925-935 - Abstract
Abstract Background The Fc glycosylation of immunoglobulin G (IgG) is well known to associate with rheumatoid arthritis (RA) disease activity. The same may be true for other classes of Igs. In the present study, we sought to determine whether the glycosylation of IgA was different between healthy subjects and patients with RA, as well as whether it was associated with RA disease activity, in particular with the pregnancy-associated improvement thereof or the flare after delivery. Methods A recently developed high-throughput method for glycoprofiling of IgA1 was applied to affinity-captured IgA from sera of patients with RA (n = 252) and healthy control subjects (n = 32) collected before, during and after pregnancy. Results IgA1 O-glycans bore more sialic acids in patients with RA than in control subjects. In addition, levels of bisecting N-acetylglucosamine of the N-glycans at asparagine 144 were higher in the patients with RA. The levels of several N-glycosylation traits were shown to change with pregnancy, similar to what has been shown before for IgG. However, the changes in IgA glycosylation were not associated with improvement or a flare of disease activity. Conclusions The glycosylation of IgA differs between patients with RA and healthy control subjects. However, our data suggest only a minor, if any, association of IgA glycosylation with RA disease activity.
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- 2017
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4. Sialic acid linkage differentiation of glycopeptides using capillary electrophoresis – electrospray ionization – mass spectrometry
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Guinevere S. M. Kammeijer, Bas C. Jansen, Isabelle Kohler, Anthonius A. M. Heemskerk, Oleg A. Mayboroda, Paul J. Hensbergen, Julie Schappler, and Manfred Wuhrer
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Medicine ,Science - Abstract
Abstract Sialylation is a glycosylation feature that occurs in different linkages at the non-reducing end of a glycan moiety, the linkage isomers are often differentially associated with various biological processes. Due to very similar physico-chemical properties, the separation of isomeric sialylated glycopeptides remains challenging but of utmost importance in the biomedicine and biotechnology, including biomarker discovery, glyco-engineering and biopharmaceutical characterization. This study presents the implementation of a high-resolution separation platform based on capillary electrophoresis – mass spectrometry (CE–MS) allowing for the selective analysis of α2,3- and α2,6-sialylated glycopeptides. These differentially linked glycopeptides showed an identical fragmentation pattern (collision induced dissociation) but different electrophoretic mobilities, allowing for baseline separation of the different linkages without the need for an extensive sample preparation. The different migration behavior between the two moieties was found to correlate with differences in pKa values. Using a novel methodology adapted from the so-called internal standard CE approach, a relative difference of 3.4·10−2 in pKa unit was determined. This approach was applied for the analysis of tryptic glycopeptides of prostate specific antigen, which shows highly complex and heterogeneous glycosylation. The developed platform therefore appears attractive for the identification of differentially linked sialic acids that may be related to pathological conditions.
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- 2017
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5. Galactosylation and Sialylation Levels of IgG Predict Relapse in Patients With PR3-ANCA Associated Vasculitis
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Michael J. Kemna, Rosina Plomp, Pieter van Paassen, Carolien A.M. Koeleman, Bas C. Jansen, Jan G.M.C. Damoiseaux, Jan Willem Cohen Tervaert, and Manfred Wuhrer
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PR3-ANCA associated vasculitis ,Granulomatosis with Polyangiitis ,Glycosylation ,Glycopeptide ,Immunoglobulin ,Mass spectrometry ,Medicine ,Medicine (General) ,R5-920 - Abstract
Objective: The objective of our study is to investigate the Fc glycosylation profiles of both antigen-specific IgG targeted against proteinase 3 (PR3-ANCA) and total IgG as prognostic markers of relapse in patients with Granulomatosis with Polyangiitis (GPA). Methods: Seventy-five patients with GPA and a PR3-ANCA rise during follow-up were included, of whom 43 patients relapsed within a median period of 8 (2–16) months. The N-glycan at Asn297 of affinity-purified and denatured total IgG and PR3-ANCA was determined by mass spectrometry of glycopeptides in samples obtained at the time of the PR3-ANCA rise and at the time of the relapse or time-matched during remission. Results: Patients with total IgG1 exhibiting low galactosylation or low sialylation were highly prone to relapse after an ANCA rise (HR 3.46 [95%-CI 1.73–6.96], p
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- 2017
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6. Serum N-Glycomic Biomarkers Predict Treatment Escalation in Inflammatory Bowel Disease
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Archana Shubhakar, Bas C Jansen, Alex T. Adams, Karli R. Reiding, Nicholas T. Ventham, Rahul Kalla, Daniel Bergemalm, Paulina A Urbanowicz, Richard A Gardner, IBD-BIOM Consortium, Jonas Halfvarson, Jack Satsangi, Daryl L Fernandes, Manfred Wuhrer, and Daniel I R Spencer
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A blood-based prognostic biomarker to guide clinical decision-making at diagnosis of inflammatory bowel disease (IBD) would be immensely helpful. We investigated a composite serum N-glycomic biomarker to predict future disease course in 244 newly diagnosed IBD patients. Forty-seven individual glycan peaks were analysed using ultra-high performance liquid chromatography identifying 105 glycoforms from which 24 derived glycan traits were calculated. Multivariable logistic regression was performed to determine associations of derived glycan traits with disease. Cox proportional hazard models were used to predict treatment escalation from first-line treatment to biologics or surgery (hazard ratio (HR) 25.9, p = 1.1×10− 12; 95% confidence interval (CI), 8.52–78.78). Application to an independent replication cohort of 54 IBD patients yielded a HR of 5.1 (p = 1.1×10− 5; 95% CI, 2.54–10.1). These data demonstrate the predictive capacity of serum N-glycan biomarkers and represent a step towards personalized medicine in IBD.
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- 2021
7. A semi-automated, high throughput approach for O-glycosylation profiling of in vitro established cancer cell lines by MALDI-FT-ICR MS
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Maximilianos, Kotsias, Katarina, Madunić, Simone, Nicolardi, Radoslaw P, Kozak, Richard A, Gardner, Bas C, Jansen, Daniel I R, Spencer, and Manfred, Wuhrer
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Glycosylation ,Polysaccharides ,Neoplasms ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Humans ,Cell Line - Abstract
The study of protein O-glycosylation is important in biological research as O-glycans have been reported to regulate a multitude of molecular and cell biology processes occurring in cancer. It is known that alterations in O-glycosylation are involved in the development and progression of cancer. Their easy accessibility makes in vitro established cell lines suitable and useful models for studying biological mechanisms in disease. However, the O-glycosylation analysis of large numbers of samples, as required in systems biology and biomarker discovery studies, is often challenging. In the present study, O-glycans from three human colorectal cancer cell lines and two human pancreatic cancer cell lines were released by semi-automated, high throughput reductive β-elimination and analysed using ultrahigh resolution MALDI-FT-ICR MS. Automated data integration and processing was performed using MassyTools, where the analyte was automatically included for relative quantitation based on a range of selection criteria including signal-to-noise ratio, mass error and isotopic pattern quality scores. A total of 126 O-glycan compositions, ranging from a single monosaccharide to large oligosaccharides exhibiting complex glycan motifs, were detected. The use of ultrahigh resolution MALDI-FTICR MS enabled glycan identification and quantitation in the matrix region of the spectrum. This approach has the potential to be used for O-glycosylation analysis of large numbers of samples, such as patient sample cohorts.
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- 2021
8. Interlaboratory evaluation of plasma N-glycan antennary fucosylation as a clinical biomarker for HNF1A-MODY using liquid chromatography methods
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Katharine R. Owen, Agata Juszczak, Haiyang Wu, Daniel Demus, Bas C. Jansen, Daniel I. R. Spencer, Nathalie Juge, Tamara Štambuk, Olga Gornik, Paulina A. Urbanowicz, Richard A. Gardner, and Edita Pape Medvidović
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Adult ,Male ,Glycan ,Glycosylation ,Fucosylation ,N-glycans ,Biology ,Biochemistry ,Maturity onset diabetes of the young ,03 medical and health sciences ,Young Adult ,Polysaccharides ,Blood plasma ,medicine ,HNF1A-MODY ,Humans ,glycosylation ,biomarker ,diagnostics ,fucosylation ,Hepatocyte Nuclear Factor 1-alpha ,Molecular Biology ,Diagnostics ,030304 developmental biology ,chemistry.chemical_classification ,Observer Variation ,0303 health sciences ,030302 biochemistry & molecular biology ,Area under the curve ,Cell Biology ,Biomarker ,medicine.disease ,Molecular biology ,3. Good health ,HNF1A ,chemistry ,Diabetes Mellitus, Type 2 ,Gene Expression Regulation ,Mutation ,biology.protein ,Biomarker (medicine) ,Female ,Original Article ,Glycoprotein ,Laboratories ,Biomarkers - Abstract
Antennary fucosylation alterations in plasma glycoproteins have been previously proposed and tested as a biomarker for differentiation of maturity onset diabetes of the young (MODY) patients carrying a functional mutation in the HNF1A gene. Here, we developed a novel LC-based workflow to analyze blood plasma N-glycan fucosylation in 320 diabetes cases with clinical features matching those at risk of HNF1A-MODY. Fucosylation levels measured in two independent research centers by using similar LC-based methods were correlated to evaluate the interlaboratory performance of the biomarker. The interlaboratory study showed good correlation between fucosylation levels measured for the 320 cases in the two centers with the correlation coefficient (r) of up to 0.88 for a single trait A3FG3S2. The improved chromatographic separation allowed the identification of six single glycan traits and a derived antennary fucosylation trait that were able to differentiate individuals carrying pathogenic mutations from benign or no HNF1A mutation cases, as determined by the area under the curve (AUC) of up to 0.94. The excellent (r = 0.88) interlaboratory performance of the glycan biomarker for HNF1A-MODY further supports the development of a clinically relevant diagnostic test measuring antennary fucosylation levels. Supplementary Information The online version contains supplementary material available at 10.1007/s10719-021-09992-w.
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- 2021
9. OP0295 N-LINKED GLYCANS IN THE VARIABLE DOMAIN OF ACPA-IGG IN THE DEVELOPMENT OF RHEUMATOID ARTHRITIS
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Lise Hafkenscheid, Xiaobo Meng, Thomas Huizinga, Manfred Wuhrer, Stacy Tanner, René E. M. Toes, Irene Smolik, Hani El-Gabalawy, Hans Scherer, Bas C. Jansen, Emma C. de Moel, and Albert Bondt
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musculoskeletal diseases ,0301 basic medicine ,Glycosylation ,Inflammatory arthritis ,Population ,Arthritis ,medicine.disease_cause ,Autoimmunity ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,skin and connective tissue diseases ,education ,030203 arthritis & rheumatology ,education.field_of_study ,Predictive marker ,biology ,business.industry ,medicine.disease ,030104 developmental biology ,chemistry ,Rheumatoid arthritis ,Immunology ,biology.protein ,Antibody ,business - Abstract
Background Anti-citrullinated protein antibodies (ACPA) are disease-specific biomarkers in rheumatoid arthritis (RA). Recently, we described that more than 90% of ACPA-IgGs harbour N-linked glycans in the antibody variable (V) domain. The corresponding N-glycosylation sites in the amino acid backbone of ACPA V-regions result from somatic hypermutation, a T cell-dependent process. Notably, both genetic evidence and data obtained from the analysis of serum ACPA indicate that T-cells drive the maturation of the ACPA-response prior to the onset of arthritis. Objectives We investigated whether ACPA-IgG carry V-domain N-glycans prior to the development of arthritis and whether the occurrence of such glycans predicts the transition from pre-disease autoimmunity to overt RA. Methods Two independent sets of serum samples were obtained from RA patients and from ACPA-positive first-degree relatives (FDR) of RA-patients (n=126) of an Indigenous North American (INA) population with high incidence rates of ACPA-positive RA. These samples comprised cross-sectional and longitudinal samples of individuals who did or did not transition to inflammatory arthritis. Serum ACPA-IgG were affinity-purified and subjected to enzymatic glycan release and UHPLC-based glycan analysis. Results ACPA-IgG V-domain glycosylation could be detected in RA patients and in FDR of RA patients. In both datasets, FDR-derived ACPA-IgG displayed markedly lower levels of V-domain glycans ( Conclusion Glycosylation of the ACPA-IgG V-domain can be detected prior to the onset of disease. Extensive glycosylation is present in a subset of predisposed FDRs of INA RA patients. The presence of this feature substantially increases the risk of RA development. These observations fit well with a pivotal role for T cells in the selection and expansion of ACPA-expressing B cells, possibly by facilitating the introduction of N-glycosylation sites in ACPA-IgG V-domains. Moreover, glycosylation of the ACPA-IgG V-domain represents a predictive marker for RA development in ACPA-positive individuals and may serve to better time and target preventive therapeutic interventions. Disclosure of Interests Lise Hafkenscheid: None declared, Emma C. de Moel: None declared, Irene Smolik: None declared, Xiaobo Meng: None declared, Stacy Tanner: None declared, Bas C. Jansen: None declared, Albert Bondt: None declared, Manfred Wuhrer: None declared, Thomas Huizinga Consultant for: Merck, UCB, Bristol Myers Squibb, Biotest AG, Pfizer, GSK, Novartis, Roche, Sanofi-Aventis, Abbott, Crescendo Bioscience Inc., Nycomed, Boeringher, Takeda, Zydus, Epirus, Eli Lilly, Rene Toes Grant/research support from: Sanofi, Hani El-Gabalawy: None declared, Hans Ulrich Scherer Grant/research support from: Sanofi, BMS
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- 2019
10. Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice
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Jona Freysdottir, Yoann Rombouts, Karli R. Reiding, Manfred Wuhrer, Andreea Ioan-Facsinay, Agnes L. Hipgrave Ederveen, Hulda S. Jónasdóttir, Ingibjorg Hardardottir, Martin Giera, Bas C. Jansen, and [ 1 ] Leiden Univ, Med Ctr, Ctr Prote & Metab, Leiden, Netherlands [ 2 ] Leiden Univ, Dept Rheumatol, Med Ctr, Leiden, Netherlands [ 3 ] Univ Toulouse, CNRS, UPS, Inst Pharmacol & Biol Struct, Toulouse, France [ 4 ] Univ Iceland, Fac Med, Biomed Ctr, Sch Hlth Sci, Reykjavik, Iceland [ 5 ] Landspitali Natl Univ Hosp Iceland, Dept Immunol, Reykjavik, Iceland [ 6 ] Landspitali Natl Univ Hosp Iceland, Ctr Rheumatol Res, Reykjavik, Iceland [ 7 ] Vrije Univ Amsterdam, Div BioAnalyt Chem, Amsterdam, Netherlands
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0301 basic medicine ,AAI12 ,Glycosylation ,Mouse ,Peritonitis ,Inflammation ,Biochemistry ,Peritoneal fluid ,Acetylglucosamine ,03 medical and health sciences ,chemistry.chemical_compound ,Plasma ,Mice ,Congenital Disorders of Glycosylation ,Zymosan-induced peritonitis ,N-linked glycosylation ,RHE12 ,N-glycosylation changes ,medicine ,Animals ,Ascitic Fluid ,Acute-Phase Reaction ,Molecular Biology ,Fucosylation ,Glycoproteins ,chemistry.chemical_classification ,Zymosan ,Acute-phase protein ,Cell Biology ,medicine.disease ,3. Good health ,carbohydrates (lipids) ,Mice, Inbred C57BL ,030104 developmental biology ,chemistry ,Original Article ,Female ,medicine.symptom ,Glycoprotein ,NAF12 ,Protein Processing, Post-Translational - Abstract
To access publisher's full text version of this article, please click on the hyperlink in Additional Links field or click on the hyperlink at the top of the page marked Files. This article is open access. Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. Importantly, substantial changes in glycosylation were already observed 12 h after induction of peritonitis, thereby demonstrating an unexpected velocity of the biological mechanisms involved. Dutch Arthritis Association (Reumafonds) LLP-24 Innovative Medicines Initiative Joint Undertaking (IMI JU)/ 115142-2 Netherlands Genomic Initiative/93511033 info:eu-repo/grantAgreement/EC/FP7/278535 info:eu-repo/grantAgreement/EC/FP7/278535
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- 2016
11. HappyTools: A software for high-throughput HPLC data processing and quantitation
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Manfred Wuhrer, Lise Hafkenscheid, Jenifer L. Hendel, Daniel I. R. Spencer, Bas C. Jansen, Albert Bondt, and Richard A. Gardner
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Data Analysis ,0301 basic medicine ,Glycosylation ,Physiology ,Computer science ,Glycobiology ,Information Theory ,lcsh:Medicine ,Biochemistry ,01 natural sciences ,High-performance liquid chromatography ,Workflow ,Software ,Immune Physiology ,Medicine and Health Sciences ,Post-Translational Modification ,lcsh:Science ,Throughput (business) ,Chromatography, High Pressure Liquid ,Liquid Chromatography ,Data processing ,Data Processing ,Immune System Proteins ,Multidisciplinary ,Chromatographic Techniques ,Software Engineering ,Biopharmaceutical ,Engineering and Technology ,Information Technology ,Research Article ,Computer and Information Sciences ,Immunology ,Research and Analysis Methods ,Antibodies ,03 medical and health sciences ,Humans ,Signal to Noise Ratio ,Data curation ,Software Tools ,business.industry ,Background Signal Noise ,010401 analytical chemistry ,lcsh:R ,Biology and Life Sciences ,Proteins ,High Performance Liquid Chromatography ,Source Code ,0104 chemical sciences ,030104 developmental biology ,Embedded system ,Signal Processing ,lcsh:Q ,business - Abstract
High-performance liquid chromatography (HPLC) is widely used for absolute quantitation. The advent of new columns and HPLC technology has enabled higher sample throughput, and hence, larger scale studies that perform quantitation on different sample types (e.g. healthy controls vs. patients with rheumatoid arthritis) using HPLC are becoming feasible. However, there remains a lack of methods that can analyse the increased number of HPLC samples. To address this in part, the modular toolkit HappyTools has been developed for the high-throughput targeted quantitation of HPLC measurements. HappyTools enables the user to create an automated workflow that includes retention time (tr) calibration, data extraction and the calculation of several quality criteria for data curation. HappyTools has been tested on a biopharmaceutical standard and previously published clinical samples. The results show comparable accuracy between HappyTools, Waters Empower and ThermoFisher Chromeleon. However, HappyTools offered superior precision and throughput when compared with Waters Empower and ThermoFisher Chromeleon. HappyTools is released under the Apache 2.0 license, both the source code and a Windows binary can be freely downloaded from https://github.com/Tarskin/HappyTools.
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- 2018
12. MIgGGly (mouse IgG glycosylation analysis) - a high-throughput method for studying Fc-linked IgG N-glycosylation in mice with nanoUPLC-ESI-MS
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Genadij Razdorov, Olga O. Zaytseva, Julija Erhardt, Maja Hanić, Mia Mrčela, Marija Pezer, Gordan Lauc, Stipan Jonjić, Ranko Stojković, Ilija Brizić, and Bas C. Jansen
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0301 basic medicine ,Analyte ,Spectrometry, Mass, Electrospray Ionization ,Glycosylation ,IgG ,Electrospray ionization ,lcsh:Medicine ,Immunoglobulin G ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,N-linked glycosylation ,Inbred strain ,Polysaccharides ,Animals ,NATURAL SCIENCES. Biology ,Effector functions ,lcsh:Science ,Biology ,Chromatography, High Pressure Liquid ,N-glycosilation ,nanoRP-LC-MS ,Mice, Inbred BALB C ,Mice, Inbred C3H ,Multidisciplinary ,PRIRODNE ZNANOSTI. Biologija ,030102 biochemistry & molecular biology ,biology ,Chemistry ,Protein ,lcsh:R ,Igg glycosylation ,Molecular biology ,Immunoglobulin Fc Fragments ,Mice, Inbred C57BL ,030104 developmental biology ,N-glycan ,biology.protein ,lcsh:Q ,Protein, N-glycosilation, N-glycan, IgG, nanoRP-LC-MS - Abstract
Immunoglobulin G (IgG) N-glycosylation is crucial for its effector functions. It is a complex trait, and large sample sets are needed to discover multiple genetic factors that underlie it. While in humans such high-throughput studies of IgG N-glycans became usual, only one has been carried out in mice. Here we describe and validate a method for the relative quantification of IgG Fc-linked N-glycans in a subclass-specific manner using nano-reverse phase liquid chromatography coupled with mass-spectrometry (nanoRP-LC-MS) applied to murine IgG. High-throughput data processing is ensured by the LaCyTools software. We have shown that IgG isolation procedure is the main source of technical variation in the current protocol. The major glycoforms were quantified reliably with coefficients of variation below 6% for all the analytes with relative abundances above 5%. We have applied our method to a sample set of 3 inbred strains: BALB/c, C57BL/6 and C3H and observed differences in subclass-specific and strain-specific N-glycosylation of IgG, suggesting a significant genetic component in the regulation of Fc-linked IgG N-glycosylation.
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- 2018
13. DOP10 Serum N-glycomic biomarkers predict treatment escalation in inflammatory bowel disease
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Manfred Wuhrer, Daryl L. Fernandes, Archana Shubhakar, Richard A. Gardner, Karli R. Reiding, Daniel I. R. Spencer, Jack Satsangi, Bas C. Jansen, R. Kalla, Daniel Bergemalm, Nicholas T. Ventham, Alex Adams, Paulina A. Urbanowicz, and Jonas Halfvarson
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medicine.medical_specialty ,business.industry ,Internal medicine ,Gastroenterology ,medicine ,General Medicine ,medicine.disease ,business ,Inflammatory bowel disease - Published
- 2019
14. MassyTools: A High-Throughput Targeted Data Processing Tool for Relative Quantitation and Quality Control Developed for Glycomic and Glycoproteomic MALDI-MS
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Manfred Wuhrer, Albert Bondt, David Falck, Agnes L. Hipgrave Ederveen, Bas C. Jansen, Karli R. Reiding, Magnus Palmblad, Rheumatology, BioAnalytical Chemistry, AIMMS, and CCA - Disease profiling
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Quality Control ,Glycan ,Glycosylation ,Bioinformatics ,relative quantitation ,Automated data processing ,Signal-To-Noise Ratio ,Biochemistry ,Software ,Polysaccharides ,Calibration ,glycoproteomics ,Humans ,Glycomics ,Profiling (computer programming) ,Background subtraction ,Data processing ,Electronic Data Processing ,Chromatography ,biology ,business.industry ,Chemistry ,matrix-assisted laser desorption/ionization ,Glycopeptides ,Antibodies, Monoclonal ,Pattern recognition ,General Chemistry ,biopharmaceuticals ,profiling mass spectrometry ,Data quality ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,biology.protein ,Artificial intelligence ,business - Abstract
The study of N-linked glycosylation has long been complicated by a lack of bioinformatics tools. In particular, there is still a lack of fast and robust data processing tools for targeted (relative) quantitation. We have developed modular, high-throughput data processing software, MassyTools, that is capable of calibrating spectra, extracting data, and performing quality control calculations based on a user-defined list of glycan or glycopeptide compositions. Typical examples of output include relative areas after background subtraction, isotopic pattern-based quality scores, spectral quality scores, and signal-to-noise ratios. We demonstrated MassyTools' performance on MALDI-TOF-MS glycan and glycopeptide data from different samples. MassyTools yielded better calibration than the commercial software flexAnalysis, generally showing 2-fold better ppm errors after internal calibration. Relative quantitation using MassyTools and flexAnalysis gave similar results, yielding a relative standard deviation (RSD) of the main glycan of similar to 6%. However, MassyTools yielded 2- to 5-fold lower RSD values for low-abundant analytes than flexAnalysis. Additionally, feature curation based on the computed quality criteria improved the data quality. In conclusion, we show that MassyTools is a robust automated data processing tool for high-throughput, high-performance glycosylation analysis. The package is released under the Apache 2.0 license and is freely available on GitHub (https://github.com/Tarskin/MassyTools ).
- Published
- 2015
15. Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3)
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Theo Rispens, Bas C. Jansen, Remco Visser, Carolien A. M. Koeleman, Gillian Dekkers, Manfred Wuhrer, Gestur Vidarsson, Paul J. Hensbergen, Rosina Plomp, Guinevere S. M. Kammeijer, Yoann Rombouts, CCA - Immuno-pathogenesis, BioAnalytical Chemistry, AIMMS, and Landsteiner Laboratory
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Adult ,Male ,Threonine ,Spectrometry, Mass, Electrospray Ionization ,Glycosylation ,Mutant ,Molecular Sequence Data ,Gene Expression ,chemical and pharmacologic phenomena ,Biology ,Biochemistry ,Immunoglobulin G ,Analytical Chemistry ,law.invention ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,law ,Tandem Mass Spectrometry ,parasitic diseases ,Humans ,Trypsin ,Amino Acid Sequence ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Research ,Proteolytic enzymes ,Middle Aged ,Recombinant Proteins ,Electron-transfer dissociation ,carbohydrates (lipids) ,chemistry ,Carbohydrate Sequence ,030220 oncology & carcinogenesis ,Monoclonal ,Mutation ,Proteolysis ,biology.protein ,Recombinant DNA ,Female ,lipids (amino acids, peptides, and proteins) ,Peptides - Abstract
Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes.
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- 2015
16. Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure
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Dietmar Reusch, Rosina Plomp, Bas C. Jansen, Manfred Wuhrer, David Falck, Markus Haberger, BioAnalytical Chemistry, AIMMS, and CCA - Disease profiling
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Proteomics ,Protein Denaturation ,Glycan ,Glycosylation ,glycosylation ,Swine ,medicine.drug_class ,CHO Cells ,Monoclonal antibody ,higher-order structure ,Biochemistry ,trypsin substrate specificity ,chemistry.chemical_compound ,immunoglobulin G ,Cricetulus ,Cricetinae ,tryptic cleavage ,medicine ,Animals ,glycoproteomics ,Trypsin ,Denaturation (biochemistry) ,Serine protease ,biology ,Glycopeptides ,Antibodies, Monoclonal ,General Chemistry ,biopharmaceuticals ,Glycoproteomics ,chemistry ,Polyclonal antibodies ,biology.protein ,monoclonal antibodies ,method development ,SDG 6 - Clean Water and Sanitation ,proteolytic biases ,medicine.drug - Abstract
It has been reported that glycosylation can influence the proteolytic cleavage of proteins. A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol.
- Published
- 2015
17. IgA N- and O-glycosylation profiling reveals no association with the pregnancy-related improvement in rheumatoid arthritis
- Author
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Simone Nicolardi, Radboud J E M Dolhain, Albert Bondt, T. Martijn Kuijper, Yuri E. M. van der Burgt, Manfred Wuhrer, Bas C. Jansen, Johanna M. W. Hazes, and Rheumatology
- Subjects
0301 basic medicine ,Immunoglobulin A ,Adult ,medicine.medical_specialty ,lcsh:Diseases of the musculoskeletal system ,Glycosylation ,Immunoglobulin G ,Arthritis, Rheumatoid ,03 medical and health sciences ,chemistry.chemical_compound ,Pregnancy ,Internal medicine ,Medicine ,Humans ,Asparagine ,Rheumatoid arthritis ,biology ,business.industry ,Healthy subjects ,medicine.disease ,Rheumatology ,3. Good health ,carbohydrates (lipids) ,Pregnancy Complications ,030104 developmental biology ,chemistry ,Immunology ,biology.protein ,Female ,lcsh:RC925-935 ,business ,Research Article - Abstract
Background The Fc glycosylation of immunoglobulin G (IgG) is well known to associate with rheumatoid arthritis (RA) disease activity. The same may be true for other classes of Igs. In the present study, we sought to determine whether the glycosylation of IgA was different between healthy subjects and patients with RA, as well as whether it was associated with RA disease activity, in particular with the pregnancy-associated improvement thereof or the flare after delivery. Methods A recently developed high-throughput method for glycoprofiling of IgA1 was applied to affinity-captured IgA from sera of patients with RA (n = 252) and healthy control subjects (n = 32) collected before, during and after pregnancy. Results IgA1 O-glycans bore more sialic acids in patients with RA than in control subjects. In addition, levels of bisecting N-acetylglucosamine of the N-glycans at asparagine 144 were higher in the patients with RA. The levels of several N-glycosylation traits were shown to change with pregnancy, similar to what has been shown before for IgG. However, the changes in IgA glycosylation were not associated with improvement or a flare of disease activity. Conclusions The glycosylation of IgA differs between patients with RA and healthy control subjects. However, our data suggest only a minor, if any, association of IgA glycosylation with RA disease activity. Electronic supplementary material The online version of this article (doi:10.1186/s13075-017-1367-0) contains supplementary material, which is available to authorized users.
- Published
- 2017
18. Galactosylation and Sialylation Levels of IgG Predict Relapse in Patients With PR3-ANCA Associated Vasculitis
- Author
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Manfred Wuhrer, Jan Damoiseaux, Rosina Plomp, Jan Willem Cohen Tervaert, Bas C. Jansen, Pieter van Paassen, Michael J. Kemna, Carolien A. M. Koeleman, Promovendi CD, MUMC+: MA Nefrologie (9), MUMC+: MA Klinische Immunologie (9), RS: NUTRIM - R3 - Respiratory & Age-related Health, RS: MHeNs - R1 - Cognitive Neuropsychiatry and Clinical Neuroscience, MUMC+: DA CDL Algemeen (9), RS: NUTRIM - R4 - Gene-environment interaction, and Faculteit FHML Centraal
- Subjects
0301 basic medicine ,ANCA-ASSOCIATED VASCULITIS ,Glycosylation ,lcsh:Medicine ,Immunoglobulin G ,chemistry.chemical_compound ,immune system diseases ,Proteinase 3 ,Recurrence ,Fucosylation ,lcsh:R5-920 ,biology ,General Medicine ,PR3-ANCA associated vasculitis ,GLYCOSYLATION CHANGES ,3. Good health ,ANTIINFLAMMATORY ACTIVITY ,Rheumatoid arthritis ,Antibody ,Granulomatosis with polyangiitis ,lcsh:Medicine (General) ,ANTIBODY-ASSOCIATED VASCULITIS ,Systemic vasculitis ,Research Paper ,Immunoglobulins ,SYSTEMIC VASCULITIS ,Glycopeptide ,General Biochemistry, Genetics and Molecular Biology ,Antibodies, Antineutrophil Cytoplasmic ,Autoimmune Diseases ,03 medical and health sciences ,MEDIATED GLOMERULONEPHRITIS ,WEGENERS-GRANULOMATOSIS ,medicine ,Immunoglobulin ,Humans ,cardiovascular diseases ,Mass spectrometry ,business.industry ,lcsh:R ,Granulomatosis with Polyangiitis ,MASS-SPECTROMETRY ,medicine.disease ,RHEUMATOID-ARTHRITIS ,carbohydrates (lipids) ,030104 developmental biology ,chemistry ,Immunology ,biology.protein ,IMMUNOGLOBULIN-G ,business - Abstract
Objective The objective of our study is to investigate the Fc glycosylation profiles of both antigen-specific IgG targeted against proteinase 3 (PR3-ANCA) and total IgG as prognostic markers of relapse in patients with Granulomatosis with Polyangiitis (GPA). Methods Seventy-five patients with GPA and a PR3-ANCA rise during follow-up were included, of whom 43 patients relapsed within a median period of 8 (2–16) months. The N-glycan at Asn297 of affinity-purified and denatured total IgG and PR3-ANCA was determined by mass spectrometry of glycopeptides in samples obtained at the time of the PR3-ANCA rise and at the time of the relapse or time-matched during remission. Results Patients with total IgG1 exhibiting low galactosylation or low sialylation were highly prone to relapse after an ANCA rise (HR 3.46 [95%-CI 1.73–6.96], p, Highlights • Low galactosylation and sialyation in total IgG predicts disease reactivation in GPA patients who experience an ANCA rise • Total IgG changes towards a more inflammation-associated phenotype in relapsing patients but not in non-relapsing patients • Glycosylation profiles may be useful in pre-emptive therapy studies using ANCA rises as guideline Patients with Granulomatosis with Polyangiitis suffer from (life-threatening) symptoms which can be kept under control using immunosuppressive therapy. However, these symptoms return in some patients, for reasons we do not yet fully understand. The disease is most likely caused by an antibody, a protein that is normally made in response to a foreign substance but can also be targeted against parts of our own body. We questioned whether there are differences in the antibodies that cause symptoms to return in some patients but not in all patients. To do this, within a group of patients in which the disease was inactive we investigated a glycan, a type of sugar group, that is attached to immunoglobulin G, the most common class of antibody. We found that the composition of this glycan showed variation in the number of certain monosaccharides, such as galactose and sialic acids. Patients in which the antibodies carried less galactose or sialic acid were more likely to suffer from symptoms of disease reactivation in the future. Moreover, the amount of sialic acid and galactose decreased over time in patients that developed symptoms, but not in patients for which the disease remained dormant. These findings may in the future be used to monitor people with inactive disease, so that timely treatment can be started if the disease becomes active.
- Published
- 2017
19. Sialic acid linkage differentiation of glycopeptides using capillary electrophoresis – electrospray ionization – mass spectrometry
- Author
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Isabelle Kohler, Oleg A. Mayboroda, Manfred Wuhrer, Anthonius A. M. Heemskerk, Guinevere S. M. Kammeijer, Paul J. Hensbergen, Bas C. Jansen, and Julie Schappler
- Subjects
0301 basic medicine ,Spectrometry, Mass, Electrospray Ionization ,Glycan ,Glycosylation ,Collision-induced dissociation ,Electrospray ionization ,Science ,Mass spectrometry ,01 natural sciences ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Capillary electrophoresis ,Carbohydrate Conformation ,ddc:615 ,Multidisciplinary ,biology ,010401 analytical chemistry ,Glycopeptides ,Electrophoresis, Capillary ,N-Acetylneuraminic Acid ,0104 chemical sciences ,Sialic acid ,030104 developmental biology ,chemistry ,Biochemistry ,biology.protein ,Medicine ,N-Acetylneuraminic acid - Abstract
Sialylation is a glycosylation feature that occurs in different linkages at the non-reducing end of a glycan moiety, the linkage isomers are often differentially associated with various biological processes. Due to very similar physico-chemical properties, the separation of isomeric sialylated glycopeptides remains challenging but of utmost importance in the biomedicine and biotechnology, including biomarker discovery, glyco-engineering and biopharmaceutical characterization. This study presents the implementation of a high-resolution separation platform based on capillary electrophoresis – mass spectrometry (CE–MS) allowing for the selective analysis of α2,3- and α2,6-sialylated glycopeptides. These differentially linked glycopeptides showed an identical fragmentation pattern (collision induced dissociation) but different electrophoretic mobilities, allowing for baseline separation of the different linkages without the need for an extensive sample preparation. The different migration behavior between the two moieties was found to correlate with differences in pKa values. Using a novel methodology adapted from the so-called internal standard CE approach, a relative difference of 3.4·10−2 in pKa unit was determined. This approach was applied for the analysis of tryptic glycopeptides of prostate specific antigen, which shows highly complex and heterogeneous glycosylation. The developed platform therefore appears attractive for the identification of differentially linked sialic acids that may be related to pathological conditions.
- Published
- 2017
20. Mo1764 – Serum N-Glycomic Biomarkers Predict Treatment Escalation in Inflammatory Bowel Disease
- Author
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Richard G. Gardner, Manfred Wuhrer, Daryl L. Fernandes, Archana Shubhakar, Nicholas T. Ventham, Bas C. Jansen, Bergemalm Daniel, Paulina A. Urbanowicz, Jack Satsangi, Alex Adams, Jonas Halfvarson, Karli R. Reiding, and Daniel I. R. Spencer
- Subjects
medicine.medical_specialty ,Hepatology ,business.industry ,Internal medicine ,Gastroenterology ,medicine ,business ,medicine.disease ,Inflammatory bowel disease - Published
- 2019
21. High-Throughput Analysis of IgG Fc Glycopeptides by LC-MS
- Author
-
David, Falck, Bas C, Jansen, Noortje, de Haan, and Manfred, Wuhrer
- Subjects
Protein Denaturation ,Glycosylation ,Immunoglobulin G ,Glycopeptides ,Humans ,Glycomics ,Chromatography, Affinity ,Mass Spectrometry ,Software ,Chromatography, Liquid ,High-Throughput Screening Assays ,Immunoglobulin Fc Fragments - Abstract
This chapter contains a nanoscale liquid chromatography-mass spectrometry method for the glycoform profiling of the conserved Fc N-glycosylation site of monoclonal and polyclonal immunoglobulin G (IgG). It describes in detail LaCyTools, a program for automated data (pre-)processing of the obtained LC-MS data. The minimal sample preparation necessary is explained as well as an optional method for affinity purification of (polyclonal) antibodies from serum or plasma.After (optional) affinity purification, the pure IgG is cleaved with trypsin. The tryptic glycopeptides are separated almost exclusively on their peptide backbone. This ensures similar response factors for all glycoforms in the MS detection and allows the collection of separate glycoform profiles for different IgG isoforms or allotypes. LaCyTools automatically performs label-free (relative) quantitation of the obtained data after minimal manual input and additionally calculates several quality criteria which can be used for data curation at the level of both individual analytes and entire LC-MS runs.
- Published
- 2016
22. High-Throughput Analysis of IgG Fc Glycopeptides by LC-MS
- Author
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Manfred Wuhrer, David Falck, Bas C. Jansen, and Noortje de Haan
- Subjects
0301 basic medicine ,Analyte ,Chromatography ,biology ,Chemistry ,010401 analytical chemistry ,01 natural sciences ,Immunoglobulin G ,0104 chemical sciences ,Glycoproteomics ,03 medical and health sciences ,Label-free quantification ,030104 developmental biology ,Affinity chromatography ,Liquid chromatography–mass spectrometry ,Polyclonal antibodies ,biology.protein ,Bottom-up proteomics - Abstract
This chapter contains a nanoscale liquid chromatography-mass spectrometry method for the glycoform profiling of the conserved Fc N-glycosylation site of monoclonal and polyclonal immunoglobulin G (IgG). It describes in detail LaCyTools, a program for automated data (pre-)processing of the obtained LC-MS data. The minimal sample preparation necessary is explained as well as an optional method for affinity purification of (polyclonal) antibodies from serum or plasma.After (optional) affinity purification, the pure IgG is cleaved with trypsin. The tryptic glycopeptides are separated almost exclusively on their peptide backbone. This ensures similar response factors for all glycoforms in the MS detection and allows the collection of separate glycoform profiles for different IgG isoforms or allotypes. LaCyTools automatically performs label-free (relative) quantitation of the obtained data after minimal manual input and additionally calculates several quality criteria which can be used for data curation at the level of both individual analytes and entire LC-MS runs.
- Published
- 2016
23. MALDI-TOF-MS reveals differential N-linked plasma- and IgG-glycosylation profiles between mothers and their newborns
- Author
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Manfred Wuhrer, Reiding Karli Robert, Gestur Vidarsson, Sicco A. Scherjon, Albert Bondt, Bas C. Jansen, Landsteiner Laboratory, and Reproductive Origins of Adult Health and Disease (ROAHD)
- Subjects
0301 basic medicine ,Glycosylation ,FC-GAMMA-RIII ,NORMAL-PREGNANCY ,Umbilical cord ,Article ,Immunoglobulin G ,Andrology ,03 medical and health sciences ,chemistry.chemical_compound ,PROTEIN-TRANSPORT ,Immune system ,BINDING ,medicine ,Receptor ,FETAL ,Fucosylation ,Pregnancy ,Fetus ,PLACENTAL TRANSPORT ,Multidisciplinary ,RECEPTOR ,biology ,Chemistry ,SIALIC-ACID ESTERIFICATION ,medicine.disease ,carbohydrates (lipids) ,030104 developmental biology ,medicine.anatomical_structure ,Immunology ,biology.protein ,IMMUNOGLOBULIN-G ,IMMUNE-SYSTEM ,lipids (amino acids, peptides, and proteins) - Abstract
During pregnancy, the mother provides multiple nutrients and substances to the foetus, with maternal immunoglobulin G (IgG) being actively transported to the foetus. Newborns depend on maternal IgG for immune-protection in their first months. The glycosylation of IgG has been shown to influence its dynamics, e.g. receptor binding. While minor differences in IgG glycosylation have been found between IgG derived from maternal blood and umbilical cord blood (UC) of newborn children, the differential glycosylation of maternal and UC plasma has hitherto not been studied. Here, we studied the N-glycosylation of IgG and total plasma proteome of both maternal and UC plasma of 42 pairs of mothers and newborn children. A total of 37 N-glycans were quantified for IgG and 45 for the total plasma N-glycome (TPNG). The study showed slightly higher levels of galactosylation for UC IgG than maternal IgG, confirming previous results, as well as lower bisection and sialylation. Furthermore, the TPNG results showed lower values for galactosylation and sialylation and higher values for fucosylation in the UC plasma. In conclusion, this study presents some novel insights into IgG glycosylation differences as well as the first broad overview of the differential plasma glycosylation between mothers and newborns.
- Published
- 2016
24. Variation of Human Salivary O-Glycome
- Author
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Radoslaw P. Kozak, Paulina A. Urbanowicz, Louise Royle, Bas C. Jansen, Chamindie Punyadeera, Manfred Wuhrer, Daryl L. Fernandes, Reiding Karli Robert, and Daniel I. R. Spencer
- Subjects
0301 basic medicine ,Saliva ,Glycosylation ,Physiology ,Glycobiology ,lcsh:Medicine ,Biochemistry ,Mass Spectrometry ,chemistry.chemical_compound ,fluids and secretions ,Medicine and Health Sciences ,Longitudinal Studies ,Biomarker discovery ,Post-Translational Modification ,lcsh:Science ,Fetuins ,Chromatography, High Pressure Liquid ,Flow Rate ,chemistry.chemical_classification ,Multidisciplinary ,biology ,Physics ,Classical Mechanics ,Hematology ,Reference Standards ,Body Fluids ,Blood ,Physical Sciences ,Metabolome ,Anatomy ,Salivation ,Research Article ,Glycan ,Electrospray ionization ,Fluid Mechanics ,Procainamide ,Continuum Mechanics ,03 medical and health sciences ,stomatognathic system ,Polysaccharides ,Animals ,Humans ,Glycoproteins ,Staining and Labeling ,lcsh:R ,Mucin ,Reproducibility of Results ,Biology and Life Sciences ,Proteins ,Fluid Dynamics ,Glycome ,carbohydrates (lipids) ,030104 developmental biology ,chemistry ,biology.protein ,lcsh:Q ,Cattle ,Glycoprotein ,Physiological Processes ,Blood Groups ,Biomarkers - Abstract
The study of saliva O-glycosylation is receiving increasing attention due to the potential of glycans for disease biomarkers, but also due to easy access and non-invasive collection of saliva as biological fluid. Saliva is rich in glycoproteins which are secreted from the bloodstream or produced by salivary glands. Mucins, which are highly O-glycosylated proteins, are particularly abundant in human saliva. Their glycosylation is associated with blood group and secretor status, and represents a reservoir of potential disease biomarkers. This study aims to analyse and compare O-glycans released from whole human mouth saliva collected 3 times a day from a healthy individual over a 5 days period. O-linked glycans were released by hydrazinolysis, labelled with procainamide and analysed by ultra-high performance liquid chromatography with fluorescence detection (UHPLC-FLR) coupled to electrospray ionization mass spectrometry (ESI-MS/MS). The sample preparation method showed excellent reproducibility and can therefore be used for biomarker discovery. Our data demonstrates that the O-glycosylation in human saliva changes significantly during the day. These changes may be related to changes in the salivary concentrations of specific proteins.
- Published
- 2016
25. LaCyTools: A Targeted Liquid Chromatography-Mass Spectrometry Data Processing Package for Relative Quantitation of Glycopeptides
- Author
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Agnes L. Hipgrave Ederveen, Manfred Wuhrer, Bas C. Jansen, David Falck, Genadij Razdorov, Noortje de Haan, and Gordan Lauc
- Subjects
0301 basic medicine ,Proteomics ,Analyte ,relative quantitation ,alpha-1-antitrypsin ,User defined ,Mass spectrometry ,01 natural sciences ,Biochemistry ,bioinformatics ,glycoproteomics ,immunoglobulin G ,liquid chromatography ,mass spectrometry ,profiling ,quality control ,Mass Spectrometry ,03 medical and health sciences ,Software ,Liquid chromatography–mass spectrometry ,Data processing ,Electronic Data Processing ,Chromatography ,Chemistry ,business.industry ,010401 analytical chemistry ,Glycopeptides ,General Chemistry ,0104 chemical sciences ,Glycoproteomics ,030104 developmental biology ,business ,Modular software ,Chromatography, Liquid - Abstract
Bottom-up glycoproteomics by liquid chromatography-mass spectrometry (LC-MS) is an established approach for assessing glycosylation in a protein- and site-specific manner. Consequently, tools are needed to automatically align, calibrate, and integrate LC-MS glycoproteomics data. We developed a modular software package designed to tackle the individual aspects of an LC-MS experiment, called LaCyTools. Targeted alignment is performed using user defined m/z and retention time (t(r)) combinations. Subsequently, sum spectra are created for each user defined analyte group. Quantitation is performed on the sum spectra, where each user defined analyte can have its own tr, minimum, and maximum charge states. Consequently, LaCyTobls deals with multiple charge states, which gives an output per charge state if desired, and offers various analyte and spectra quality criteria. We compared throughput and performance of LaCyTools to combinations of available tools that deal with individual processing steps. LaCyTools yielded relative quantitation of equal precision (relative standard deviation
- Published
- 2016
26. Dopant Enriched Nitrogen Gas Combined with Sheathless Capillary Electrophoresis-Electrospray Ionization-Mass Spectrometry for Improved Sensitivity and Repeatability in Glycopeptide Analysis
- Author
-
Isabelle Kohler, Oleg A. Mayboroda, Paul J. Hensbergen, Manfred Wuhrer, David Falck, Guinevere S. M. Kammeijer, and Bas C. Jansen
- Subjects
0301 basic medicine ,Detection limit ,Spectrometry, Mass, Electrospray Ionization ,Chromatography ,Dopant ,Chemistry ,Nitrogen ,Electrospray ionization ,010401 analytical chemistry ,Analytical chemistry ,Glycopeptides ,Electrophoresis, Capillary ,Repeatability ,Mass spectrometry ,01 natural sciences ,0104 chemical sciences ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,Capillary electrophoresis ,Ionization ,Acetonitrile ,Chromatography, Liquid - Abstract
Over the last years, numerous strategies have been proposed to enhance both ionization efficiency and spray stability in electrospray ionization (ESI), in particular for nanospray applications. In nano-liquid chromatography-mass spectrometry (nano-LC-ESI-MS), a better ESI performance has been observed when a coaxial gas flow is added around the ESI emitter. Moreover, enrichment of the gas with an organic dopant has led to an improved desolvation and ionization efficiency with an overall enhanced sensitivity. In this study, the use of a dopant enriched nitrogen (DEN)-gas combined with sheathless capillary electrophoresis (CE)-ESI-MS was evaluated for glycopeptide analysis. Using acetonitrile as a dopant, an increased sensitivity was observed compared to conventional sheathless CE-ESI-MS. Up to 25-fold higher sensitivities for model glycopeptides were obtained, allowing for limits of detection unachieved by state-of-the-art nano-LC-ESI-MS. The effect of DEN-gas on the repeatability and intermediate precision was also investigated. When compared to previously reported nano-LC-ESI-MS measurements, similar values were found for CE-ESI-MS with DEN-gas. The enhanced repeatability fosters the use of DEN-gas sheathless CE-ESI-MS in protein glycosylation analysis, where precision is essential. The use of DEN-gas opens new avenues for highly sensitive sheathless CE-ESI-MS approaches in glycoproteomics research, by significantly improving sensitivity and precision.
- Published
- 2016
27. Human plasma protein N-glycosylation
- Author
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Manfred Wuhrer, Albert Bondt, Bas C. Jansen, Guinevere S. M. Kammeijer, Florent Clerc, and Karli R. Reiding
- Subjects
0301 basic medicine ,Serum ,Apolipoprotein D ,Glycan ,Glycosylation ,Immunoglobulins ,N-glycosylation ,Biology ,Biochemistry ,03 medical and health sciences ,chemistry.chemical_compound ,Plasma ,N-linked glycosylation ,Serotransferrin ,Humans ,Molecular Biology ,Glycoproteins ,chemistry.chemical_classification ,Hemopexin ,Blood Proteins ,Cell Biology ,Glycoproteomics ,3. Good health ,carbohydrates (lipids) ,030104 developmental biology ,chemistry ,biology.protein ,Original Article ,lipids (amino acids, peptides, and proteins) ,Glycoprotein ,Protein Processing, Post-Translational - Abstract
Glycosylation is the most abundant and complex protein modification, and can have a profound structural and functional effect on the conjugate. The oligosaccharide fraction is recognized to be involved in multiple biological processes, and to affect proteins physical properties, and has consequentially been labeled a critical quality attribute of biopharmaceuticals. Additionally, due to recent advances in analytical methods and analysis software, glycosylation is targeted in the search for disease biomarkers for early diagnosis and patient stratification. Biofluids such as saliva, serum or plasma are of great use in this regard, as they are easily accessible and can provide relevant glycosylation information. Thus, as the assessment of protein glycosylation is becoming a major element in clinical and biopharmaceutical research, this review aims to convey the current state of knowledge on the N-glycosylation of the major plasma glycoproteins alpha-1-acid glycoprotein, alpha-1-antitrypsin, alpha-1B-glycoprotein, alpha-2-HS-glycoprotein, alpha-2-macroglobulin, antithrombin-III, apolipoprotein B-100, apolipoprotein D, apolipoprotein F, beta-2-glycoprotein 1, ceruloplasmin, fibrinogen, immunoglobulin (Ig) A, IgG, IgM, haptoglobin, hemopexin, histidine-rich glycoprotein, kininogen-1, serotransferrin, vitronectin, and zinc-alpha-2-glycoprotein. In addition, the less abundant immunoglobulins D and E are included because of their major relevance in immunology and biopharmaceutical research. Where available, the glycosylation is described in a site-specific manner. In the discussion, we put the glycosylation of individual proteins into perspective and speculate how the individual proteins may contribute to a total plasma N-glycosylation profile determined at the released glycan level. Electronic supplementary material The online version of this article (doi:10.1007/s10719-015-9626-2) contains supplementary material, which is available to authorized users.
- Published
- 2016
28. Pregnancy-associated serum N-glycome changes studied by high-throughput MALDI-TOF-MS
- Author
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Karli R. Reiding, Guinevere S. M. Kammeijer, Albert Bondt, Yoann Rombouts, Coen J. de Jong, Radboud J E M Dolhain, Manfred Wuhrer, Bas C. Jansen, David Falck, Emanuela Lonardi, and Rheumatology
- Subjects
Serum ,0301 basic medicine ,Glycan ,Time Factors ,Glycosylation ,Polysaccharide ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Immune system ,Polysaccharides ,Pregnancy ,medicine ,Humans ,chemistry.chemical_classification ,Electronic Data Processing ,Multidisciplinary ,biology ,medicine.disease ,Glycome ,Healthy Volunteers ,3. Good health ,Sialic acid ,carbohydrates (lipids) ,Matrix-assisted laser desorption/ionization ,030104 developmental biology ,chemistry ,Biochemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,biology.protein ,Female - Abstract
Pregnancy requires partial suppression of the immune system to ensure maternal-foetal tolerance. Protein glycosylation and especially terminal sialic acid linkages, are of prime importance in regulating the pro- and anti-inflammatory immune responses. However, little is known about pregnancy-associated changes of the serum N-glycome and sialic acid linkages. Using a combination of recently developed methods, i.e. derivatisation that allows the distinction between α2,3- and α2,6-linked sialic acids by high-throughput MALDI-TOF-MS and software-assisted data processing, we analysed the serum N-glycome of a cohort of 29 healthy women at 6 time points during and after pregnancy. A total of 77 N-glycans were followed over time, confirming in part previous findings while also revealing novel associations (e.g. an increase of FA2BG1S1(6), FA2G1S1(6) and A2BG2S2(6) with delivery). From the individual glycans we calculated 42 derived traits. With these, an increase during pregnancy and decrease after delivery was observed for both α2,3- and α2,6-linked sialylation. Additionally, a difference in the recovery speed after delivery was observed for α2,3- and α2,6-linked sialylation of triantennary glycans. In conclusion, our new high-throughput workflow allowed the identification of novel plasma glycosylation changes with pregnancy.
- Published
- 2016
- Full Text
- View/download PDF
29. Longitudinal monitoring of immunoglobulin A glycosylation during pregnancy by simultaneous MALDI-FTICR-MS analysis of N- and O-glycopeptides
- Author
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Guinevere S. M. Kammeijer, Kathrin Stavenhagen, Bas C. Jansen, Paul J. Hensbergen, Albert Bondt, Johanna M. W. Hazes, Yuri E. M. van der Burgt, Dennis Blank, Simone Nicolardi, Manfred Wuhrer, Daryl L. Fernandes, Radboud J E M Dolhain, Radoslaw P. Kozak, and Rheumatology
- Subjects
0301 basic medicine ,Immunoglobulin A ,Glycosylation ,Mass spectrometry ,Article ,Immunoglobulin G ,Fourier transform ion cyclotron resonance ,03 medical and health sciences ,chemistry.chemical_compound ,Polysaccharides ,Pregnancy ,Spectroscopy, Fourier Transform Infrared ,Humans ,Immunologic Factors ,Longitudinal Studies ,chemistry.chemical_classification ,Multidisciplinary ,biology ,Glycopeptide ,3. Good health ,Matrix-assisted laser desorption/ionization ,030104 developmental biology ,chemistry ,Biochemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Immunology ,biology.protein ,Female ,Glycoprotein - Abstract
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.
- Published
- 2016
30. Variation of Human Salivary O-Glycome.
- Author
-
Radoslaw P Kozak, Paulina A Urbanowicz, Chamindie Punyadeera, Karli R Reiding, Bas C Jansen, Louise Royle, Daniel I Spencer, Daryl L Fernandes, and Manfred Wuhrer
- Subjects
Medicine ,Science - Abstract
The study of saliva O-glycosylation is receiving increasing attention due to the potential of glycans for disease biomarkers, but also due to easy access and non-invasive collection of saliva as biological fluid. Saliva is rich in glycoproteins which are secreted from the bloodstream or produced by salivary glands. Mucins, which are highly O-glycosylated proteins, are particularly abundant in human saliva. Their glycosylation is associated with blood group and secretor status, and represents a reservoir of potential disease biomarkers. This study aims to analyse and compare O-glycans released from whole human mouth saliva collected 3 times a day from a healthy individual over a 5 days period. O-linked glycans were released by hydrazinolysis, labelled with procainamide and analysed by ultra-high performance liquid chromatography with fluorescence detection (UHPLC-FLR) coupled to electrospray ionization mass spectrometry (ESI-MS/MS). The sample preparation method showed excellent reproducibility and can therefore be used for biomarker discovery. Our data demonstrates that the O-glycosylation in human saliva changes significantly during the day. These changes may be related to changes in the salivary concentrations of specific proteins.
- Published
- 2016
- Full Text
- View/download PDF
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