Review
Chemistry, Analytical
Denisa Smolkova, Richard Cmelik, Jana Lavicka
Summary: Capillary electrophoresis is widely used for oligosaccharide and glycan analysis, but the low electrophoretic mobility and lack of chromophore or fluorophore in saccharide molecules require modification before separation and detection. This review summarizes the labeling strategies used for oligosaccharide/glycan analysis, categorized based on the labeling mechanism and saccharide molecule. In addition to the commonly used 8-aminopyrene-1,3,6-trisulfonic acid trisodium salt, less common and recently developed tags applicable in glycomics are also summarized.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2023)
Article
Chemistry, Analytical
Jieqiang Zhong, Yifan Huang, Yehia Mechref
Summary: This study presents an efficient derivatization method for discriminating sialic acid linkage isomers on glycopeptides, resulting in increased signal and successful characterization of sialylated glycopeptides from various proteins. The method targets site- and sialic acid linkage-specific profiling of glycosylation, making it a facile and highly reproducible technique.
ANALYTICAL CHEMISTRY
(2021)
Article
Biochemical Research Methods
Hua Zhang, Xudong Shi, Yuan Liu, Bin Wang, Meng Xu, Nathan Welham, Lingjun Li
Summary: Spatial visualization of N-glycans in clinical tissue samples is achieved through a novel on-tissue derivatization strategy. This strategy utilizes the amidation reaction between sialic acid residues and aniline to stabilize and enhance the signal intensity of sialylated N-glycans. Proof-of-concept experiments and mass spectrometry imaging demonstrate the improved detection sensitivity and characterization of N-glycan dysregulation in cancer tissue samples.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Alan B. Moran, Georgia Elgood-Hunt, Yuri E. M. van der Burgt, Manfred Wuhrer, Wilma E. Mesker, Rob A. E. M. Tollenaar, Daniel I. R. Spencer, Guinevere S. M. Lageveen-Kammeijer
Summary: A newly developed analytical strategy was used to analyze the N-glycome of colorectal cancer (CRC) patients before and after surgery. The strategy involved fluorescent labeling and specific derivatization techniques, allowing for efficient separation and detection. The results showed changes in N-glycan structures, which may have implications for CRC classification and clinical data analysis. This strategy can be used to monitor patient profiles during and after clinical events and warrants further investigation.
Article
Chemistry, Analytical
Yiting Wang, Yang Hu, Ru Wang, Wei Zhang, Huiting Mao, Chuanjun Yuan, Ruinian Hua
Summary: In this study, a highly sensitive non-proteinuria assay platform was developed using urinary sialic acid as a biomarker to accurately diagnose diabetic nephropathy within a short time period. This platform incorporated the ninhydrin reaction and the inner filter effect, resulting in stable and high sensitivity with 100% positive prediction for urine samples.
Article
Chemistry, Analytical
Yiting Wang, Yang Hu, Ru Wang, Wei Zhang, Huiting Mao, Chuanjun Yuan, Ruinian Hua
Summary: Diabetic nephropathy (DN), the most common microvascular complication associated with incurable diabetes, can now be accurately diagnosed within 20 minutes using an ultrasensitive non-proteinuria assay platform. This platform utilizes urinary sialic acid as an effective biomarker for DN and combines mimetic immunoassay and inner filter effect to convert imperceptible color into highly sensitive fluorescence signals.
Article
Chemistry, Analytical
Jieqiang Zhong, Yifan Huang, Peilin Jiang, Yehia Mechref
Summary: Glycosylation is a crucial post-translational modification. However, the characterization of negatively charged sialoglycopeptides associated with various diseases is challenging due to co-existence with high abundant peptides and low ionization efficiency. Therefore, an efficient enrichment method for sialoglycopeptides is essential. This study presents a novel derivatization-based enrichment method that identifies linkage isomers of sialic acids, unifies the net charge of peptides, and introduces positive charges to sialoglycopeptides. The method enables efficient enrichment through electrostatic interaction using weak cation exchange (WCX) media.
ANALYTICA CHIMICA ACTA
(2022)
Article
Chemistry, Analytical
Taiga Iwama, Kuniyuki Kano, Daisuke Saigusa, Kim Ekroos, Gerhild van Echten-Deckert, Johannes Vogt, Junken Aoki
Summary: The novel on-tissue derivatization method using Phos-tag in combination with MALDI-MSI enables visualization of low-abundance bioactive lipids like LPA and S1P in the brain, providing a simple and optimal way to reveal the spatial localization of these important molecules in tissues.
ANALYTICAL CHEMISTRY
(2021)
Article
Chemistry, Analytical
Mengxia Cheng, Hong Shu, Ye Peng, Xiaoxiao Feng, Guoquan Yan, Lei Zhang, Jun Yao, Huimin Bao, Haojie Lu
Summary: A comprehensive strategy combining linkage-specific derivatization and charge-sensitive separation method was developed for specific analysis of alpha-2,3-sialylated N-glycan linkage isomers. This new approach demonstrated successful sensitive profiling and reproducible quantitation of serum alpha-2,3-sialylated N-glycome from ovarian cancer patients, identifying significant changes in certain glycan structures.
ANALYTICAL CHEMISTRY
(2021)
Article
Biochemical Research Methods
Taylor M. Domenick, Austin L. Jones, Robin H. J. Kemperman, Richard A. Yost
Summary: To address the poor retention of highly polar metabolites in reversed-phase liquid chromatography (RPLC), a derivatization agent and an internal standard were introduced to improve analyte retention and detection limits. Multiplexed drift-tube ion mobility (IM) was integrated into the LC-MS workflow to enhance method sensitivity and selectivity. The results showed that this method allowed for the quantitation of amino acids in microglial cell extracts.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Analytical
Ge-Ge Gong, Jie Zheng, Sha Li, Ya-Li Bai, Yu-Qi Feng
Summary: A LC-MS method was developed to simultaneously determine 5 RAs in human serum, with enhanced analysis throughput and increased detection sensitivity. The concentrations of three RAs in cancer patient serum samples were significantly decreased compared with health controls, indicating a potential role of RAs in cancer formation and development.
Article
Chemistry, Analytical
Ling Ling, Shaoning Yu, Chuanfan Ding
Summary: Direct analysis of glycans by MALDI MS has limited sensitivity, but using 4-HQ as a reactive matrix can improve the detection sensitivity with good reproducibility and simple application.
Review
Chemistry, Multidisciplinary
Dylan Nicholas Tabang, Megan Ford, Lingjun Li
Summary: Protein modification by glycans is crucial for mediating biological functions, and mass spectrometry (MS) has become a powerful tool for glycomic and glycoproteomic analyses in various diseases. Recent advances in pancreatic disease research using MS have led to the discovery of new biomarkers and understanding of the links between glycans and disease pathology.
FRONTIERS IN CHEMISTRY
(2021)
Article
Chemistry, Applied
Taylor E. Gray, Kamal Narayana, Alexander M. Garner, Samantha A. Bakker, Rachael K. H. Yoo, Amanda J. Fischer-Tlustos, Michael A. Steele, Wesley F. Zandberg
Summary: The study confirmed the existence of sulfated and sialylated BMOs and their direct biosynthetic competition, as well as identified over 170 unique BMOs, including 14 unique glucuronic acid-containing trisaccharides.
Article
Biochemical Research Methods
Yuan Zhi, Li Jia, Jiechen Shen, Jun Li, Zexuan Chen, Bojing Zhu, Zhifang Hao, Yintai Xu, Shisheng Sun
Summary: Formic acid solution is commonly used for dissolving glycopeptide and glycan samples before mass spectrometry analysis. However, storing the samples in formic acid solution at -20°C can lead to an undesirable modification that affects the identification and quantification of glycans. Therefore, storing samples at -20°C should be avoided.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2022)
Article
Biochemical Research Methods
Baubek Spanov, Bas Baartmans, Oladapo Olaleye, Simone Nicolardi, Natalia Govorukhina, Manfred Wuhrer, Nico C. van de Merbel, Rainer Bischoff
Summary: We have developed a method using pH gradient cation-exchange chromatography to reveal the charge heterogeneity of stressed trastuzumab at the subunit level. This approach can be used for charge variant analysis of therapeutic antibodies with two or more modification sites in the Fab region.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2023)
Article
Chemistry, Analytical
Steffen Lippold, Michaela Hook, Christian Spick, Alexander Knaupp, Kevin Whang, Fabian Ruperti, Lance Cadang, Nisana Andersen, Annette Vogt, Michael Grote, Dietmar Reusch, Markus Haberger, Feng Yang, Tilman Schlothauer
Summary: T-cell engaging bispecific antibodies (TCBs) targeting CD3 and tumor-specific antigens show great promise as therapeutics. In this study, we developed a novel method using functional CD3 target affinity chromatography (AC) coupled with online mass spectrometry (MS) to separate and identify critical modified proteoforms of TCBs. This method enables the functional distinction of important modifications and the evaluation of product-related variants, streamlining the integration of functional and structural characterizations of therapeutic CD3 targeting TCBs.
ANALYTICAL CHEMISTRY
(2023)
Article
Immunology
Nienke Oskam, Timon Damelang, Marij Streutker, Pleuni Ooijevaar-de Heer, Jan Nouta, Carolien Koeleman, Julie Van Coillie, Manfred Wuhrer, Gestur Vidarsson, Theo Rispens
Summary: Of the four human IgG subclasses, IgG4 is the least inflammatory due to poor activation of the complement system. However, in certain diseases, IgG4 antibodies have been linked to complement consumption and deposition. This study investigated the ability of IgG4 to activate complement and found that it can only do so at high antigen and antibody concentrations via the classical pathway. Glycosylation and Fab arm exchange influenced complement activation, with reduced Fc galactosylation diminishing activation. These findings suggest that IgG4-mediated complement activation in pathology is possible but unlikely.
FRONTIERS IN IMMUNOLOGY
(2023)
Article
Medicine, General & Internal
Julie Van Coillie, Tamas Pongracz, Johann Rahmoeller, Hung-Jen Chen, Chiara Elisabeth Geyer, Lonneke A. van Vught, Jana Sophia Buhre, Tonci Sustic, Thijs Luc Junior van Osch, Maurice Steenhuis, Willianne Hoepel, Wenjun Wang, Anne Sophie Lixenfeld, Jan Nouta, Sofie Keijzer, Federica Linty, Remco Visser, Mads Delbo Larsen, Emily Lara Martin, Inga Kuensting, Selina Lehrian, Vera von Kopylow, Carsten Kern, Hanna Bele Lunding, Menno de Winther, Niels van Mourik, Theo Rispens, Tobias Graf, Marleen Adriana Slim, Rene Peter Minnaar, Marije Kristianne Bomers, Jonne Jochum Sikkens, Alexander P. J. Vlaar, C. Ellen van der Schoot, Jeroen den Dunnen, Manfred Wuhrer, Marc Ehlers, Gestur Vidarsson
Summary: The BNT162b2 mRNA vaccine induces transient afucosylated anti-S IgG1 responses in naive individuals, but not in antigen-experienced ones. Further studies are needed to determine the clinical context in which potent afucosylated responses would be preferred.
Article
Chemistry, Analytical
Guusje van Schaick, Elena Dominguez-Vega, Jerome Castel, Manfred Wuhrer, Oscar Hernandez-Alba, Sarah Cianferani
Summary: Post-translational modifications (PTMs) increase protein structural heterogeneity and can alter conformation and biological functions. Monitoring PTMs is crucial for therapeutic products like monoclonal antibodies (mAbs). Cation-exchange chromatography (CEX) with native mass spectrometry is valuable for characterizing mAb charge variants, but cannot explore protein conformation. To address this, we combined CEX separation with collision-induced unfolding (CIU) experiments to monitor conformational differences without compromising CEX resolution. This strategy enabled in-depth characterization of mAb glycoforms and linking conformational changes to resistance against gas-phase unfolding charge variants.
ANALYTICAL CHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Di Wang, Valeriia Kuzyk, Katarina Madunic, Tao Zhang, Oleg A. Mayboroda, Manfred Wuhrer, Guinevere S. M. Lageveen-Kammeijer
Summary: Colorectal cancer (CRC) is a common and deadly type of cancer. Studying the N-glycosylation of CRC cell lines can help identify potential therapeutic and diagnostic targets. The study found a diverse range of N-glycans among the examined CRC cell lines and discovered similarities between different platforms used for analysis.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Agriculture, Multidisciplinary
Rui-Rui Guo, Guinevere S. M. Lageveen-Kammeijer, Wenjun Wang, Hans Dalebout, Wangang Zhang, Manfred Wuhrer, Li Liu, Bram Heijs, Josef Voglmeir
Summary: Severe allergic reactions to certain types of meat have been observed in areas with ticks, due to an immune response to a carbohydrate antigen called alpha-Gal. The presence and distribution of alpha-Gal-modified N-glycans in different meats were studied, and it was found that these modified glycans were abundant in beef, mutton, and pork, mainly present in fibroconnective tissue. This study contributes to a better understanding of the glycosylation biology of meat and provides guidance for processed meat products.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2023)
Article
Biochemical Research Methods
Alan B. Moran, Elena Dominguez-Vega, Manfred Wuhrer, Guinevere S. M. Lageveen-Kammeijer
Summary: Intact protein analysis by mass spectrometry is important for assessing post-translational modifications and biotransformation. The lack of consensus on the best practice for data processing and unfit data processing tools make analysis of intact proteins laborious. This study investigated a software-assisted workflow for streamlined intact protein data processing and quantification. The results showed comparable quantification to manually performed quantification, demonstrating the efficiency of the presented workflow.
JOURNAL OF PROTEOME RESEARCH
(2023)
Article
Cell Biology
Ruiyao Xu, Xianxian Wang, Sadia Safi, Nico Braunegger, Agnes Hipgrave Ederveen, Michelle Rottmann, Joachim Wittbrodt, Manfred Wuhrer, Janine Wesslowski, Gary Davidson
Summary: Reception of Wnt signals is mediated by Frizzled receptors and a co-receptor LRP6 or LRP5. Precise control of receptor activation is crucial to regulate Wnt/beta-catenin signaling for proper cellular function. A glycosyltransferase B3GnT2-like has been identified to modify LRP6 and enhance Wnt/beta-catenin signaling. This study highlights the importance of LRP6 as a regulatory hub in Wnt signaling and provides insight into the selective targeting of a signaling pathway component by a specific glycosyltransferase.
Article
Biochemistry & Molecular Biology
Alan B. Moran, Georgia Elgood-Hunt, Yuri E. M. van der Burgt, Manfred Wuhrer, Wilma E. Mesker, Rob A. E. M. Tollenaar, Daniel I. R. Spencer, Guinevere S. M. Lageveen-Kammeijer
Summary: A newly developed analytical strategy was used to analyze the N-glycome of colorectal cancer (CRC) patients before and after surgery. The strategy involved fluorescent labeling and specific derivatization techniques, allowing for efficient separation and detection. The results showed changes in N-glycan structures, which may have implications for CRC classification and clinical data analysis. This strategy can be used to monitor patient profiles during and after clinical events and warrants further investigation.
Article
Endocrinology & Metabolism
Elham Memarian, Ralph Heijmans, Roderick C. Slieker, Adriana Sierra, Olga Gornik, Joline W. J. Beulens, Maja Hanic, Petra Elders, Julio Pascual, Eric Sijbrands, Gordan Lauc, Viktoria Dotz, Clara Barrios, Leen M. 't Hart, Manfred Wuhrer, Mandy van Hoek
Summary: Inflammation plays an important role in the development of type 2 diabetes complications, and the N-glycosylation of IgG affects its role in inflammation. This study found that IgG N-glycosylation, particularly galactosylation and to a lesser extent sialylation, is associated with a higher prevalence and future development of macro- and microvascular complications of diabetes.
DIABETES-METABOLISM RESEARCH AND REVIEWS
(2023)
Article
Chemistry, Applied
Guusje van Schaick, Sanne Pot, Olaf Schouten, Joost den Hartog, Michiel Akeroyd, Rob van der Hoeven, Wim Bijleveld, Nicolas Abello, Manfred Wuhrer, Maurien Olsthoorn, Elena Dominguez-Vega
Summary: Protein glycation is a natural process in industrial enzymes and its impact on enzyme performance is unpredictable. A boronate affinity chromatography method was developed to analyze the influence of glycation on enzyme activity and stability. The results showed that glycation did not affect the performance of the lipase.
Article
Biochemistry & Molecular Biology
Annemieke Naber, Daniel Demus, Roderick Slieker, Simone Nicolardi, Joline W. J. Beulens, Petra J. M. Elders, Aloysius G. Lieverse, Eric J. G. Sijbrands, Leen M. 't Hart, Manfred Wuhrer, Mandy van Hoek
Summary: This study aimed to investigate the impact of apo-CIII glycosylation on lipid traits and type 2 diabetes prevalence. The researchers conducted a GWAS on different apo-CIII proteoforms and identified genetic loci associated with glycosylation and its relation to lipids and diabetes. The study confirmed the significant role of GALNT2 gene in apo-CIII O-glycosylation and its association with lipid parameters. Additionally, the IFT172/NRBP1 region was found to be involved in apo-CIII sialylation and hypertriglyceridemia.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemical Research Methods
Florent Clerc, Karli R. Reiding, Noortje de Haan, Carolien A. M. Koeleman, Agnes L. Hipgrave Ederveen, Natalia Manetti, Viktoria Dotz, Vito Annese, Manfred Wuhrer
Summary: This study investigates the associations of immunoglobulin A1 and A2 glycosylation with inflammatory bowel diseases (IBD). Differences in IgA O- and N-glycosylation between patient groups were found to be associated with the diseases, and a statistical model was constructed to predict the disease group of the patients. These findings contribute to the development of noninvasive biomarkers for IBD and improvement in patient care.
JOURNAL OF PROTEOME RESEARCH
(2023)
Article
Biochemistry & Molecular Biology
Oleg A. Mayboroda, Guinevere S. M. Lageveen-Kammeijer, Manfred Wuhrer, Radboud J. E. M. Dolhain
Summary: Rheumatoid arthritis (RA) is a prevalent autoimmune disease affecting joints and other organs. Glycosylation changes in immunoglobulin G (IgG) and immunoglobulin A (IgA) have been found in RA patients, which can be used as biomarkers to differentiate patients and predict disease activity.
