A5023111668- Mass Spectrometry Techniques and Applications
- Advanced Proteomics Techniques and Applications
- Glycosylation and Glycoproteins Research
- Metabolomics and Mass Spectrometry Studies
- Monoclonal and Polyclonal Antibodies Research
- Protein purification and stability
- Analytical Chemistry and Chromatography
- RNA and protein synthesis mechanisms
- Carbohydrate Chemistry and Synthesis
- Microfluidic and Capillary Electrophoresis Applications
- Bacterial Genetics and Biotechnology
- Polysaccharides and Plant Cell Walls
- Lipid metabolism and disorders
- Gut microbiota and health
- Pharmaceutical and Antibiotic Environmental Impacts
- Advanced Cellulose Research Studies
- Bacterial Identification and Susceptibility Testing
- Pancreatic and Hepatic Oncology Research
- Pancreatitis Pathology and Treatment
- Enzyme Production and Characterization
- Diabetes and associated disorders
- Biofuel production and bioconversion
- Genomics and Phylogenetic Studies
- Molecular Biology Techniques and Applications
- Diabetes, Cardiovascular Risks, and Lipoproteins
Netherlands Metabolomics Centre
2013-2024
Leiden University Medical Center
2015-2024
Leiden University
2017-2018
University of Pisa
2017
Sapienza University of Rome
2007
MALDI mass spectrometry imaging is able to simultaneously determine the spatial distribution of hundreds molecules directly from tissue sections, without labeling and prior knowledge. Ultra-high resolution measurements based on Fourier-transform have been utilized resolve isobaric lipids, metabolites tryptic peptides. Here we demonstrate potential 15T MALDI-FTICR MSI for molecular pathology in a mouse model high-grade glioma. The high accuracy resolving power field FTICR enabled tumor...
The Consortium for Top-Down Proteomics (www.topdownproteomics.org) launched the present study to assess current state of top-down mass spectrometry (TD MS) and middle-down (MD characterizing monoclonal antibody (mAb) primary structures, including their modifications. To meet needs rapidly growing therapeutic market, it is important develop analytical strategies characterize heterogeneity a product's structure accurately reproducibly. major objective determine whether TD/MD MS technologies...
Abstract Glycans are involved in various life processes and represent critical targets of biomedical developments. Nevertheless, the accessibility to long glycans with precise structures remains challenging. Here we report on synthesis consisting [→4)-α-Rha-(1 → 3)-β-Man-(1 ] repeating unit, which relevant O -antigen Bacteroides vulgatus , a common component gut microbiota. The optimal combination assembly strategy, protecting group arrangement, glycosylation reaction has enabled us...
The emergence of complex protein therapeutics in general and monoclonal antibodies (mAbs) particular have stimulated analytical chemists to develop new methods strategies for their structural characterization. Mass spectrometry plays a key role providing information on the primary amino acid sequence, post-translational modifications, other structure characteristics that must be monitored during manufacturing process subsequent quality control assessment. In this study, we present novel...
An accurate mass measurement of a known protein provides information on potential amino acid deletions and post-translational modifications. Although this field is dominated by strategies based electrospray ionization, spectrometry (MS) methods using matrix-assisted laser desorption/ionization (MALDI) have the advantage yielding predominantly singly charged precursor ions, thus avoiding peak overlap from different charge states multiple species. Such MALDI-MS require at ultrahigh resolution,...
High-throughput mass spectrometry (MS) glycomics is an emerging field driven by technological advancements including sample preparation and data processing. Previously, we reported automated protocol for the analysis of N-glycans released from plasma proteins that included sialic acid derivatization with linkage-specificity, namely, ethylation α2,6-linked residues lactone formation α2,3-linked acids. In current study, each step in this was further optimized. Method improvements minimizing...
Biofunctionalized gold nanoparticles (AuNPs) enable innovative translational research and development in biomedicine. Biomolecules such as peptides, proteins, lipids, carbohydrates can be assembled onto AuNPs to yield nanomaterials with unique properties for applications imaging, photothermal therapy, vaccination strategies, drug delivery. The characterization of functionalized still remains an analytical challenge that normally requires the combination multiple techniques. Laser...
Determination of the impact individual antibody glycoforms on FcɣRIIIa affinity, and consequently antibody-dependent cell-mediated cytotoxicity (ADCC) previously required high purity glycoengineering. We hyphenated affinity chromatography to mass spectrometry, which allowed direct comparison intact monoclonal antibodies. The approach enabled reproduction refinement known glycosylation effects, insights afucosylation pairing as well low-abundant, unstudied glycoforms. Our method greatly...
Apolipoprotein-CIII (apo-CIII) inhibits the clearance of triglycerides from circulation and is associated with an increased risk diabetes complications. It exists in four main proteoforms: O-glycosylated variants containing either zero, one, or two sialic acids a non-glycosylated variant. O-glycosylation may affect metabolic functions apo-CIII. We investigated associations apo-CIII glycosylation blood plasma, measured by mass spectrometry intact protein, genetic micro- macrovascular...
Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation either zero, one or two sialic acids. Changes in the relative abundance of these observed variety pathologies. In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms serum protein profiles. For purpose, proteins...
Structural confirmation and quality control of recombinant monoclonal antibodies (mAbs) by top-down mass spectrometry is still challenging due to the size proteins, disulfide content, post-translational modifications such as glycosylation. In this study we have applied electrochemistry (EC) overcome bridge complexity in analysis mAbs. To end, an electrochemical cell was coupled directly electrospray ionization (ESI) source a Fourier transform ion cyclotron resonance (FTICR) spectrometer (MS)...
Bispecific monoclonal antibodies (BsAbs) are engineered proteins with multiple functionalities and properties. The "bi-specificity" of these complex biopharmaceuticals is a key characteristic for the development novel more effective therapeutic strategies. high structural complexity BsAbs poses challenge to analytical methods needed their characterization. Modifications BsAb structure, resulting from enzymatic non-enzymatic processes, further complicate analysis. An important example latter...
The asialoglycoprotein receptor (ASGPR) and the mannose C-type 1 (MRC1) are well known for their selective recognition clearance of circulating glycoproteins. Terminal galactose N-Acetylgalactosamine recognized by ASGPR, while terminal mannose, fucose, N-Acetylglucosamine MRC1. effects ASGPR MRC1 deficiency on N-glycosylation individual proteins have been studied. However, impact homeostasis major plasma glycoproteins is debated glycosylation has not mapped with high molecular resolution in...
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 high-throughput strategy the simultaneous serum-derived IgA1 -glycopeptides using matrix-assisted laser/desorption ionisation Fourier transform ion cyclotron resonance (MALDI-FTICR) mass spectrometry (MS). Six non-fucosylated...
Abstract Background &Aims Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer type with loco‐regional spread that makes the tumor surgically unresectable. Novel diagnostic tools are needed to improve detection of PDAC and increase patient survival. In this study we explore serum protein N ‐glycan profiles from patients regard their applicability serve as a disease biomarker panel. Methods Total ‐glycome analysis was applied discovery set (86 cases/84 controls) followed by...
The Fc glycosylation of immunoglobulin G (IgG) is well known to associate with rheumatoid arthritis (RA) disease activity. same may be true for other classes Igs. In the present study, we sought determine whether IgA was different between healthy subjects and patients RA, as it associated RA activity, in particular pregnancy-associated improvement thereof or flare after delivery.A recently developed high-throughput method glycoprofiling IgA1 applied affinity-captured from sera (n = 252)...
Carbohydrates, such as oligo- and polysaccharides, are highly abundant biopolymers that involved in numerous processes. The study of their structure functions is commonly based on a material isolated from complex natural sources. However, more precise analysis requires pure compounds with well-defined structures can be obtained chemical or enzymatic syntheses. Novel synthetic strategies have increased the accessibility larger monodisperse posing challenge to analytical methods used for...
Abstract Breast cancer is the most prevalent in women. Early detection of this disease improves survival and therefore population screenings, based on mammography, are performed. However, sensitivity screening modality not optimal new methods, such as blood tests, being explored. Most analyses that aim for early focus proteins bloodstream. In study, biomarker potential total serum N -glycosylation analysis was explored with regard to breast cancer. an age-matched case-control setup protein...
Abstract Background Changes in plasma protein glycosylation are known to functionally affect proteins and associate with liver diseases, including cirrhosis hepatocellular carcinoma. Autoimmune hepatitis (AIH) is a disease characterized by inflammation raised serum levels of IgG, difficult distinguish from other diseases. The aim this study was examine IgG-specific N -glycosylation AIH compare it healthy controls Methods In cross-sectional cohort study, total IgG Fc analysis performed mass...