High-Throughput Glycomic Methods

Glycomics aims to identify the structure and function of the glycome, the complete set of oligosaccharides (glycans), produced in a given cell or organism, as well as to identify genes and other factors that govern glycosylation. This challenging endeavor requires highly robust, sensitive, and potentially automatable analytical technologies for the analysis of hundreds or thousands of glycomes in a timely manner (termed high-throughput glycomics). This review provides a historic overview as well as highlights recent developments and challenges of glycomic profiling by the most prominent high-throughput glycomic approaches, with N-glycosylation analysis as the focal point. It describes the current state-of-the-art regarding levels of characterization and most widely used technologies, selected applications of high-throughput glycomics in deciphering glycosylation process in healthy and disease states, as well as future perspectives.

• Publication year

  2022

• Venue

  Chemical Reviews

• Publication date

  2022-07-07

• Fields of study

  Biology, Medicine, Materials Science, Chemistry

• Identifiers
  DOI 10.1021/acs.chemrev.1c01031PMID 35797639PMCID 9614987
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• In-Depth Profiling of O-Glycan Isomers in Human Cells Using C18 Nanoliquid Chromatography–Mass Spectrometry and Glycogenomics

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• Genetic regulation of post-translational modification of two distinct proteins

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• Fucosylated AGP glycopeptides as biomarkers of HNF1A-Maturity Onset Diabetes of the Young.

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• The minimum information required for a glycomics experiment (MIRAGE): reporting guidelines for capillary electrophoresis.

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• N-Glycosylation Patterns across the Age-Related Macular Degeneration Spectrum

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• Immunoglobulin G glycome composition in transition from premenopause to postmenopause

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• Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis.

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• Tailor-made recombinant prokaryotic lectins for characterisation of glycoproteins.

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• Small RNAs are modified with N-glycans and displayed on the surface of living cells.

  2021cited by this paper
• Sialic acid as a potential biomarker for cardiovascular disease, diabetes and cancer.

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• Direct N-Glycosylation Profiling of Urine and Prostatic Fluid Glycoproteins and Extracellular Vesicles

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• The Analysis of Human Serum N-Glycosylation in Patients with Primary and Metastatic Brain Tumors

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• Profile of Immunoglobulin G N-Glycome in COVID-19 Patients: A Case-Control Study

  2021cited by this paper
• Plasma protein N-glycosylation is associated with cardiovascular disease, nephropathy, and retinopathy in type 2 diabetes

  2021cited by this paper
• Label‐Free Protein Glycosylation Analysis Using NanoMonitor—An Ultrasensitive Electrochemical Biosensor

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• Gold standard for nutrition: a review of human milk oligosaccharide and its effects on infant gut microbiota

  2021cited by this paper
• The glycoconjugate ontology (GlycoCoO) for standardizing the annotation of glycoconjugate data and its application

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• High-throughput glycopeptide profiling of prostate-specific antigen from seminal plasma by MALDI-MS.

  2021cited by this paper
• Human milk oligosaccharides, infant growth, and adiposity over the first 4 months of lactation

  2021cited by this paper
• Longitudinal Changes in Human Milk Oligosaccharides (HMOs) Over the Course of 24 Months of Lactation.

  2021cited by this paper
• The structure and role of lactone intermediates in linkage-specific sialic acid derivatization reactions

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• Streamlined Subclass-Specific Absolute Quantification of Serum IgG Glycopeptides Using Synthetic Isotope-Labeled Standards.

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• Glycosylation and Aging.

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• Rapid-Throughput Analysis of Human Milk Oligosaccharides from Filipino Breastmilk

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• Sweet systems: technologies for glycomic analysis and their integration into systems biology

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• Discrepancies between High-Resolution Native and Glycopeptide-Centric Mass Spectrometric Approaches: A Case Study into the Glycosylation of Erythropoietin Variants

  2021cited by this paper
• Serum and Plasma Immunoglobulin G Fc N-Glycosylation Is Stable during Storage

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• High-throughput and high-sensitivity N-Glycan profiling: a platform for biopharmaceutical development and disease biomarker discovery.

  2021cited by this paper
• Glycoform analysis of intact erythropoietin by MALDI FT-ICR mass spectrometry.

  2021cited by this paper
• Afucosylated Plasmodium falciparum-specific IgG is induced by infection but not by subunit vaccination

  2021cited by this paper
• N-Glycomic Signature of Stage II Colorectal Cancer and Its Association With the Tumor Microenvironment

  2021cited by this paper
• Analysis of Monoclonal Antibodies by Capillary Electrophoresis: Sample Preparation, Separation, and Detection

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• Large-Scale Analysis of Apolipoprotein CIII Glycosylation by Ultrahigh Resolution Mass Spectrometry

  2021influential reference
• Glycosylation and Cardiovascular Diseases.

  2021cited by this paper
• Dopant-Enriched Nitrogen Gas for Enhanced Electrospray Ionization of Released Glycans in Negative Ion Mode

  2021cited by this paper
• Immunoglobulin G glycome composition in transition from pre-menopause to menopause

  2021cited by this paper
• N-glycosylation of immunoglobulin G predicts incident hypertension

  2021cited by this paper
• High-Throughput and Site-Specific N-Glycosylation Analysis of Human Alpha-1-Acid Glycoprotein Offers a Great Potential for New Biomarker Discovery

  2021cited by this paper
• Glycosylation of IgG Associates with Hypertension and Type 2 Diabetes Mellitus Comorbidity in the Chinese Muslim Ethnic Minorities and the Han Chinese

  2021cited by this paper
• Fc Galactosylation Promotes Hexamerization of Human IgG1, Leading to Enhanced Classical Complement Activation

  2021cited by this paper
• Changes of Serum IgG Glycosylation Patterns in Primary Biliary Cholangitis Patients

  2021cited by this paper
• Robustness and repeatability of GlycoWorks RapiFluor-MS IgG N-glycan profiling in a long-term high-throughput glycomic study.

  2021cited by this paper
• Global patterns of breast cancer incidence and mortality: A population‐based cancer registry data analysis from 2000 to 2020

  2021cited by this paper
• O- and N-Glycosylation of Serum Immunoglobulin A is Associated with IgA Nephropathy and Glomerular Function

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• Global epidemiology of systemic lupus erythematosus

  2021cited by this paper
• Antibody Subclass and Glycosylation Shift Following Effective TB Treatment

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• Extensive weight loss reduces glycan age by altering IgG N-glycosylation

  2021cited by this paper
• Immunoglobulin G1 Fc glycosylation as an early hallmark of severe COVID-19

  2021cited by this paper
• Comprehensive analysis of glycosphingolipid glycans by lectin microarrays and MALDI-TOF mass spectrometry

  2021cited by this paper
• Associations of Human Milk Oligosaccharides and Bioactive Proteins with Infant Morbidity and Inflammation in Malawian Mother-Infant Dyads

  2021cited by this paper
• Biology of human milk oligosaccharides: From basic science to clinical evidence

  2021cited by this paper
• Plasma N-glycome shows continuous deterioration as the diagnosis of insulin resistance approaches

  2021cited by this paper
• MS-based glycomics and glycoproteomics methods enabling isomeric characterization.

  2021cited by this paper
• N-glycosylation profiling of Type 2 diabetes mellitus from baseline to follow-up: an observational study in a Ghanaian population.

  2021cited by this paper
• GeenaR: A Web Tool for Reproducible MALDI-TOF Analysis

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• Serum N-glycan profiles differ for various breast cancer subtypes

  2021cited by this paper
• Application of Lectin Microarrays for Biomarker Discovery

  2020cited by this paper
• Glycomics studies using sialic acid derivatization and mass spectrometry

  2020cited by this paper
• Altered glycosylation associated with dedifferentiation of hepatocellular carcinoma: a lectin microarray-based study

  2020cited by this paper
• Glycosylation of immunoglobulin G is regulated by a large network of genes pleiotropic with inflammatory diseases

  2020cited by this paper
• Serum N-glycan markers for diagnosing liver fibrosis induced by hepatitis B virus

  2020cited by this paper
• rMSIproc: an R package for mass spectrometry imaging data processing

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• OUP accepted manuscript

  2020cited by this paper
• Protein Glycosylation

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• Dissecting Total Plasma and Protein-Specific Glycosylation Profiles in Congenital Disorders of Glycosylation

  2020cited by this paper
• Decreased Immunoglobulin G Core Fucosylation, A Player in Antibody-dependent Cell-mediated Cytotoxicity, is Associated with Autoimmune Thyroid Diseases*

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• Toolbox Accelerating Glycomics (TAG): Glycan Annotation from MALDI-TOF MS Spectra and Mapping Expression Variation to Biosynthetic Pathways

  2020cited by this paper
• Choosing proper normalization is essential for discovery of sparse glycan biomarkers.

  2020cited by this paper
• Ultrafast high-resolution analysis of human milk oligosaccharides by multicapillary gel electrophoresis.

  2020cited by this paper
• Detection of small molecule concentration gradients in ocular tissues and humours.

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• N-glycomic signature of stage II colorectal cancer and its association with the tumor microenvironment.

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• Simultaneous Immunoglobulin A and G Glycopeptide Profiling for High-Throughput Applications

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• Composition of the immunoglobulin G glycome associates with the severity of COVID-19

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• Clustering and curation of electropherograms: an efficient method for analyzing large cohorts of capillary electrophoresis glycomic profiles for bioprocessing operations

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• N-glycans as functional effectors of genetic and epigenetic disease risk.

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• Rapid N-Glycan Profiling of Serum and Plasma by a Novel Slide-Based Imaging Mass Spectrometry Workflow.

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• Enhanced protocol for quantitative N-linked glycomics analysis using Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tags (INLIGHT)™

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• On-Tissue Derivatization with Girard’s Reagent P Enhances N-Glycan Signals for Formalin-Fixed Paraffin-Embedded Tissue Sections in MALDI Mass Spectrometry Imaging

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• Metformin and statin use associate with plasma protein N-glycosylation in people with type 2 diabetes

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• C-Type Lectin Receptors in Host Defense Against Bacterial Pathogens

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• Human Milk Oligosaccharide profiles and allergic disease up to 18 years.

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• Immunoglobulin G Glycoprofiles are Unaffected by Common Bottom-Up Sample Processing

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• Lectin microarrays for glycoproteomics: an overview of their use and potential

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• Afucosylated IgG characterizes enveloped viral responses and correlates with COVID-19 severity

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• In situ isobaric lipid mapping by MALDI-ion mobility separation-mass spectrometry imaging.

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• Recent Advances in Analytical Approaches for Glycan and Glycopeptide Quantitation

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• High-throughput and site-specific N-glycosylation analysis of human alpha-1-acid glycoprotein offers a great potential for new biomarker discovery.

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• High-Throughput Analysis of Fluorescently Labeled N-Glycans Derived from Biotherapeutics Using an Automated LC-MS-Based Solution

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• Differential Influence on Antibody Dependent Cellular Phagocytosis by Different Glycoforms on Therapeutic Monoclonal Antibodies.

  2020cited by this paper
• Plasma N-Glycans as Emerging Biomarkers of Cardiometabolic Risk: A Prospective Investigation in the EPIC-Potsdam Cohort Study

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• O‐glycosylated clusterin as a sensitive marker for diagnosing early stages of prostate cancer

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• High-throughput glycomic analyses reveal unique oligosaccharide profiles of canine and feline milk samples

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• Multiplexed imaging mass spectrometry of the extracellular matrix using serial enzyme digests from formalin-fixed paraffin-embedded tissue sections

  2020cited by this paper
• Integrated glycomics strategy for the evaluation of glycosylation alterations in salivary proteins associated with type 2 diabetes mellitus

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• Replication of fifteen loci involved in human plasma protein N-glycosylation in 4,802 samples from four cohorts.

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• Molecular signatures of tumor progression in myxoid liposarcoma identified by N-glycan mass spectrometry imaging

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• Human milk oligosaccharide DSLNT and gut microbiome in preterm infants predicts necrotising enterocolitis

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• Recent advances in the analysis of human milk oligosaccharides by liquid phase separation methods.

  2020cited by this paper
• Plasma and antibody glycomic biomarkers of time to HIV rebound and viral setpoint.

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• New strategies for profiling and characterization of human milk oligosaccharides

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• GlycoForge generates realistic glycomics data under known ground truth for rigorous method benchmarking

  2026cites this paper
• Glycomic Insights in Gynecological Disease: From Molecular Mechanisms to Precision Diagnostics and Therapeutics

  2026cites this paper
• Machine Learning-Enhanced Analysis of Exosomal Surface Sialic Acid Using Surface-Enhanced Raman Spectroscopy for Ovarian Cancer Diagnosis and Therapeutic Monitoring.

  2026cites this paper
• GlyTrait: A Versatile Bioinformatics Tool for Glycomics Analysis.

  2025cites this paper
• Immunoglobulin G N-glycosylation predicts long-term risk of ischemic stroke: a nested case-control study

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• Compositional data analysis enables statistical rigor in comparative glycomics

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• Lectin Microarray-based Glycomics and Machine Learning Identify Shared Osteoarthritis Biomarkers in Humans, Dogs, and Horses

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• Immunoglobulin G Subclass-Specific Glycosylation Changes in Rheumatoid Arthritis

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• A systematic review of protein post-translational modifications in sepsis

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• Plasma N-Glycan Signatures Are Associated With a First Event of Venous Thromboembolism

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• Advancement in Clinical Glycomics and Glycoproteomics for Congenital Disorders of Glycosylation: Progress and Challenges Ahead

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• Boosting Quantification of N‑Glycans by an Enhanced Isobaric Multiplex Reagents for Carbonyl-Containing Compound (SUGAR) Tagging Strategy

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• Analytical dissection of minor glycoforms and glycoprotein associations in rAAV preparations by multimodal glycoproteomics

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• Advances in mass spectrometry-based glycomics: progress, applications, and perspectives.

  2025cites this paper
• Glycomics in the Clinic:A Brief Introduction toGlycosylation-Informed Medicine

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• Aberrant protein glycosylation in the colon adenoma-cancer sequence: Colorectal cancer mechanisms and clinical implications.

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• New insights into phytochemicals via protein glycosylation focused on aging and diabetes.

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• Impact of Mobile Phase Composition on Separation Selectivity of Labeled Dextran Ladder in Hydrophilic Interaction Liquid Chromatography

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• GlycoDash: automated, visually assisted curation of glycoproteomics datasets for large sample numbers

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• Swift Universal Glycan Acquisition (SUGA) Enables Quantitative Glycan Profiling across Diverse Sample Types

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• Impact of different pathogen classes on the serum N-glycome in septic shock

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• Application of foodomics technologies in the study of peanut quality: A review

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• Decoding Sugars: Mass Spectrometric Advances in the Analysis of the Sugar Alphabet

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• Absolute Quantification of Aging-Associated Glycans in IgG for Biological Age Prediction: Insights from Glycomics and Transcriptomics

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• GlycoPro: A High-Throughput Sample-Processing Platform for Multi-Glycosylation-Omics Analysis

  2025cites this paper
• Oxidative Release of Natural Glycans (ORNG): Preparation of Truncated N-glycans for Engineering Glycoproteins.

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• Advances in MS-based glycomics for biomedical research.

  2025cites this paper
• A funnel-type pipeline for accelerating discovery of tailored designer xylanosomes for efficient arabinoxylan valorization

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• Microbial Dynamics and Quality Monitoring in Biopharmaceutical Production

  2024cites this paper
• An N-glycome tissue atlas of 15 human normal and cancer tissue types determined by MALDI-imaging mass spectrometry

  2024cites this paper
• Advances in Mass Spectrometry-Based Blood Metabolomics Profiling for Non-Cancer Diseases: A Comprehensive Review

  2024cites this paper
• Influence of plasma collection tubes on N-glycome in human blood samples

  2024cites this paper
• Machine learning framework to extract the biomarker potential of plasma IgG N-glycans towards disease risk stratification

  2024cites this paper
• Decoding the glycoproteome: a new frontier for biomarker discovery in cancer

  2024cites this paper
• Exploring the significance of glycosylation in prostate cancer subtyping through single-cell analysis

  2024cites this paper
• The Human Blood N-Glycome: Unraveling Disease Glycosylation Patterns

  2024cites this paper
• Comprehensive O-Glycan Analysis by Porous Graphitized Carbon Nanoliquid Chromatography–Mass Spectrometry

  2024cites this paper
• Recent advances in N-glycan biomarker discovery among human diseases

  2024cites this paper
• Extracellular vesicles and glycans: new avenue for biomarker research

  2024influential citation
• N-Glycosylation of Plant Proteins

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• Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

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• Developing glycoproteomics reveals the role of posttranslational glycosylation in the physiological and pathological processes of male reproduction

  2024cites this paper
• 色谱在糖组学分析中的应用

  2024cites this paper
• GlyTrait Brings Insights into Functional Glycosylation

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• The glycosylation landscape of prostate cancer tissues and biofluids.

  2024cites this paper
• Fingerprint Profiling of Glycans on Extracellular Vesicles via Lectin-Induced Aggregation Strategy for Precise Cancer Diagnostics.

  2024cites this paper
• Lectin-Based Approaches to Analyze the Role of Glycans and Their Clinical Application in Disease

  2024cites this paper
• The importance of IgG glycosylation—What did we learn after analyzing over 100,000 individuals

  2024cites this paper
• High-throughput N-glycan analysis in aging and inflammaging: State of the art and future directions.

  2024cites this paper
• The general glycan profiling of Dendrobium officinale and their protective effects on MIN6 cells via ERK signaling pathway.

  2024cites this paper
• Cracking the diversity of sweet drugs

  2024cites this paper
• Labeling of oligosaccharides and N-linked glycans by a rhodamine-based fluorescent tag for analysis by capillary electrophoresis with laser-induced fluorescence and mass spectrometry detection.

  2024cites this paper
• The Blessed Union of Glycobiology and Immunology: A Marriage That Worked

  2023cites this paper
• Kütle Spektrometresinden Glikan Mikrodizilerine: Glikomikte Analitik Tekniklere Genel Bir Bakış

  2023cites this paper
• Recommendations for reproducibility of cerebrospinal fluid extracellular vesicle studies

  2023cites this paper
• Transcription Factors HNF1A, HNF4A, and FOXA2 Regulate Hepatic Cell Protein N-Glycosylation

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• Recent progress in high-throughput and automated characterization of N-glycans in monoclonal antibodies

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• Can we suppress chronic systemic inflammation and postpone age-related diseases by targeting the IgG glycome?

  2023cites this paper
• Integration of High-pH Fractionation and Cotton-HILIC Enrichment Approaches for Analyzing N-glycopeptides

  2023cites this paper
• Imaging of N-Linked Glycans in Biological Tissue Sections Using Nanospray Desorption Electrospray Ionization (nano-DESI) Mass Spectrometry.

  2023cites this paper
• The role of the glycome in symbiotic host-microbe interactions

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• Magnetic nanoagent assisted deciphering of heterogeneous glycans in extracellular vesicles of varied cellular origins.

  2023cites this paper
• Immunoglobulin A Glycosylation Differs between Crohn’s Disease and Ulcerative Colitis

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• Amyloid-β: A potential mediator of aging-related vascular pathologies.

  2023cites this paper
• Comprehensive review of MS‐based studies on N‐glycoproteome and N‐glycome of extracellular vesicles

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• Predicting glycan structure from tandem mass spectrometry via deep learning

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• Removable Dyes—The Missing Link for In-Depth N-glycan Analysis via Multi-Method Approaches

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• IgG glycans in health and disease: Prediction, intervention, prognosis, and therapy.

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• The Role of Clinical Glyco(proteo)mics in Precision Medicine

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• Potential Clinically Relevant Effects of Sialylation on Human Serum AAG-Drug Interactions Assessed by Isothermal Titration Calorimetry: Insight into Pharmacoglycomics?

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• Roles of glycosylation at the cancer cell surface: opportunities for large scale glycoproteomics

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• Warning Signs From the Crypt: Aberrant Protein Glycosylation Marks Opportunities for Early Colorectal Cancer Detection

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• Stable isotope labeling differential glycans discovery in the serum of acute myocardial infarction by ultrahigh-performance liquid chromatography-quadrupole-Orbitrap high resolution mass spectrometry.

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• Prediction of gastric cancer by machine learning integrated with mass spectrometry-based N-glycomics.

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• Critical considerations in N-glycoproteomics.

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• In a pursuit of optimal glycan fluorescent label for negative MS mode for high-throughput N-glycan analysis

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• Mixing nitrogenous ligands in osmium(VI)ligand-polysaccharide complexes. Voltammetric sensing

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• Glycoinformatics in the Artificial Intelligence Era

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