Manfred Wuhrer
A5065397150- Glycosylation and Glycoproteins Research
- Monoclonal and Polyclonal Antibodies Research
- Galectins and Cancer Biology
- Carbohydrate Chemistry and Synthesis
- Mass Spectrometry Techniques and Applications
- Advanced Proteomics Techniques and Applications
- Protein purification and stability
- T-cell and B-cell Immunology
- Blood groups and transfusion
- Analytical Chemistry and Chromatography
- Proteoglycans and glycosaminoglycans research
- Parasites and Host Interactions
- Complement system in diseases
- Immune Cell Function and Interaction
- Platelet Disorders and Treatments
- Immunodeficiency and Autoimmune Disorders
- SARS-CoV-2 and COVID-19 Research
- Genomics and Phylogenetic Studies
- Systemic Lupus Erythematosus Research
- Microfluidic and Capillary Electrophoresis Applications
- Immunotherapy and Immune Responses
- Diabetes and associated disorders
- Chronic Lymphocytic Leukemia Research
- Pancreatic and Hepatic Oncology Research
- Trypanosoma species research and implications
Netherlands Metabolomics Centre
2016-2025
Leiden University Medical Center
2016-2025
Loyola University Medical Center
2010-2025
University of Copenhagen
2024
Noguchi Memorial Institute for Medical Research
2024
Leiden University
2013-2023
Vrije Universiteit Amsterdam
2013-2019
Proteogenomics Research Institute for Systems Medicine
2013-2018
Amsterdam UMC Location Vrije Universiteit Amsterdam
2014-2016
University of Nottingham
2010
All immunoglobulin G molecules carry N-glycans, which modulate their biological activity. Changes in N-glycosylation of IgG associate with various diseases and affect the activity therapeutic antibodies intravenous immunoglobulins. We have developed a novel 96-well protein monolithic plate used it to rapidly isolate from plasma 2298 individuals three isolated human populations. N-glycans were released by PNGase F, labeled 2-aminobenzamide analyzed hydrophilic interaction chromatography...
Summary Mannose‐binding type 1 pili are important virulence factors for the establishment of Escherichia coli urinary tract infections (UTIs). These initiated by adhesion uropathogenic E. to uroplakin receptors in uroepithelium via FimH adhesin located at tips pili. Blocking bacterial is able prevent infection. Here, we provide first time binding data molecular events underlying fimbrial adherence, crystallographic analyses receptor domains from a and K‐12 strain, affinity measurements with...
Immunoglobulin G (IgG) antibodies are crucial for protection against invading pathogens. A highly conserved N-linked glycan within the IgG-Fc tail, which is essential IgG function, shows variable composition in humans. Afucosylated variants already used anticancer therapeutic their increased activity through Fc receptors (FcγRIIIa). Here, we report that afucosylated (approximately 6% of total humans) specifically formed enveloped viruses but generally not other antigens. This mediates...
Solid-phase extraction microtips are important devices in modern bioanalytics, as they allow miniaturized sample preparation for mass spectrometric analysis. Here we introduce the use of cotton wool filter-free HILIC SPE microtips. To this end, pieces pads (approximately 500 μg) were packed into 10 μL pipet tips. The performance tips was evaluated microscale purification tryptic IgG Fc N-glycopeptides. Cotton allowed removal salts, most nonglycosylated peptides, and detergents such SDS from...
Glycosylation of the immunoglobulin G (IgG)-Fc tail is required for binding to Fc-gamma receptors (FcγRs) and complement-component C1q. A variety IgG1 glycoforms detected in human sera. Several groups have found global or antigen specific skewing IgG glycosylation, example autoimmune diseases, viral infections, alloimmune reactions. The glycoprofiles seem correlate with disease outcome. Additionally, IgG-glycan composition contributes significantly Ig based therapies, as IVIg diseases...
Protein glycosylation is an important post-translational modification associated, among others, with diseases and the efficacy of biopharmaceuticals. Matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) can be performed to study in a high-throughput manner, but hampered by instability ionization bias experienced sialylated glycan species. Stabilization neutralization these sialic acids achieved permethylation or specific carboxyl group derivatization...
Current approaches to study glycosylation of polyclonal human immunoglobulins G (IgG) usually imply protein digestion or glycan release. While these allow in-depth characterization, they also result in a loss valuable information regarding certain subclasses, allotypes and co-occuring post-translational modifications (PTMs). Unfortunately, the high variability IgGs makes their intact mass spectrometry (MS) analysis extremely challenging. We propose here middle-up strategy for fragment...
Objective To determine whether anticitrullinated protein antibodies (ACPA) exhibit specific changes in Fc glycosylation prior to the onset of arthritis. Methods Serum samples patients with ACPA-positive arthralgia (n=183) were collected at baseline and various time points follow-up. 105 developed arthritis after a median 12 months (IQR 6–24) classified as having either rheumatoid (RA, n=48) or undifferentiated (UA, n=57) based on 1987 American College Rheumatology (ACR) criteria. ACPA total...
The N-linked glycosylation of the constant fragment (Fc) immunoglobulin G has been shown to change during pathological and physiological events strongly influence antibody inflammatory properties. In contrast, little is known about Fab-linked N-glycosylation, carried by ∼20% IgG. Here we present a high-throughput workflow analyze Fab Fc polyclonal IgG purified from 5 μl serum. We were able detect quantify 37 different N-glycans means MALDI-TOF-MS analysis in reflectron positive mode using...
Abstract Epigenetic alterations may provide important insights into gene-environment interaction in inflammatory bowel disease (IBD). Here we observe epigenome-wide DNA methylation differences 240 newly-diagnosed IBD cases and 190 controls. These include 439 differentially methylated positions (DMPs) 5 regions (DMRs), which study detail using whole genome bisulphite sequencing. We replicate the top DMP ( RPS6KA2 ) DMRs VMP1, ITGB2 TXK an independent cohort. Using paired genetic epigenetic...
Patients diagnosed with coronavirus disease 2019 (COVID-19) become critically ill primarily around the time of activation adaptive immune response. Here, we provide evidence that antibodies play a role in worsening at seroconversion. We show early-phase severe acute respiratory distress syndrome 2 (SARS-CoV-2) spike protein-specific immunoglobulin G (IgG) serum COVID-19 patients induces excessive inflammatory responses by human alveolar macrophages. identified this response is dependent on...
Monomeric serum immunoglobulin A (IgA) can contribute to the development of various autoimmune diseases, but regulation IgA effector functions is not well defined. Here, we show that two subclasses (IgA1 and IgA2) differ in their effect on immune cells due distinct binding signaling properties. Whereas IgA2 acts pro-inflammatory neutrophils macrophages, IgA1 does have pronounced effects. Moreover, different glycosylation profiles, with possessing more sialic acid than IgA2. Removal increases...
Antibody dependent cellular cytotoxicity (ADCC) is an Fc-dependent effector function of IgG important for anti-viral immunity and anti-tumor therapies. NK-cell mediated ADCC mainly triggered by IgG-subclasses IgG1 IgG3 through the IgG-Fc-receptor (FcγR) IIIa. Polymorphisms in immunoglobulin gamma heavy chain gene likely form a layer variation strength ADCC-response, but this has never been studied detail. We produced all 27 known allotypes assessed FcγRIIIa binding activity. While IgG1, IgG2...
Primary membranous nephropathy (pMN) is a leading cause of nephrotic syndrome in adults. In most cases, this autoimmune kidney disease associated with autoantibodies against the M-type phospholipase A2 receptor (PLA2R1) expressed on podocytes, but mechanisms to glomerular damage remain elusive. Here, we developed cell culture model using human podocytes and found that anti-PLA2R1–positive pMN patient sera or isolated IgG4, not IgG4-depleted sera, induced proteolysis 2 essential podocyte...
Background The new types of mRNA-containing lipid nanoparticle vaccines BNT162b2 and mRNA-1273 the adenovirus-based vaccine AZD1222 were developed against SARS-CoV-2 code for its spike (S) protein. Several studies have investigated short-term antibody (Ab) responses after vaccination. Objective However, impact these formats with unclear effects on long-term Ab response – including isotype, subclass, their type Fc glycosylation is less explored. Methods Here, we analyzed anti-S in blood serum...
Abstract Despite recent advances in cancer immunotherapy, pancreatic ductal adenocarcinoma (PDAC) remains unresponsive due to an immunosuppressive tumor microenvironment, which is characterized by the abundance of cancer-associated fibroblasts (CAFs). Once identified, CAF-mediated immune inhibitory mechanisms could be exploited for immunotherapy. Siglec receptors are increasingly recognized as checkpoints, and their ligands, sialic acids, known overexpressed cells. Here, we unveil a...