N-glycans are enzymatically released from glycoproteins with PNGase, labeled, purified, and analyzed by different high-resolution mass spectrometry.
Our workflows ensure consistency, reproducibility, and scalability across high sample volumes.
MALDI-TOF profiling of released N-glycans
To enhance ionization efficiency and sialic acid residues are stabilized by esterification/lactonization, enabling the mass-based differentiation of α2,3- and α2,6-linked sialic acid linkages. MALDI-TOF mass spectrometry provides a fast and cost-effective method for glycan screening, making it especially suitable for large-scale or time-sensitive projects.
UHPLC-MS profiling of released N-glycans
UHPLC-MS combines chromatographic separation with high-resolution Q-TOF analysis. Separation methods as fast as 3 minutes per sample enable quantitative profiling of typical IgG N-glycans without compromising sensitivity or accuracy, compared to longer, more time-consuming separation gradients for a fraction of the cost.
Comparative evaluation of N-glycan profiling using three distinct separation methods confirms consistent performance, maintaining sensitivity and accuracy while enabling high-throughput workflows with a reduced run time of 3 minutes.
Fast UHPLC-MS profiling of N-glycans from human plasma IgG.
How we work for HTP Profiling of N-glycans
Our workflow ensures a clear, straightforward, and transparent process for sample glycoanalysis in an HTP setup
Sample
Let us know which type of sample (purified protein, antibodies, cells, tissue, etc.) you want to analyze, and we will apply the most convenient sample preparation protocol.
Glycan release and isolation
We will use the optimal enzymes, techniques, and procedures to have the glycans ready to be analyzed.
Chemical labelling of glycans
We will choose the labeling method according to the detection techniques.
Sample purification
We enrich and purify the samples accordingly to obtain the best signals.
Mass spectrometry analysis
The latest technology and instruments are used to ensure the data quality.
Data processing
Our experts analyze and interpret the data to transform it into actionable information.
Report
A publication-type report that includes the Materials & Methods, data interpretation, and potential further analysis.
Purposes and applications for HTP of N-glycans
N-glycan population characterization of purified glycoproteins in large amounts of samples serves several purposes and applications:
Oncology
HTP N-glycan profiling rapidly identifies tumor-associated glycosylation changes across large patient cohorts, enabling biomarker validation and accelerating the development of glycomimetic cancer therapies
Infectious Diseases
Through HTP analysis, N-glycan receptors used by pathogens for entry can be systematically mapped, supporting vaccine design and high-throughput screening of antiviral candidates
Immune and Inflammatory Diseases
HTP N-glycan profiling uncovers subtle immune glycosylation changes in large sample sets, facilitating the discovery of diagnostic markers and the optimization of glycoengineered immunotherapies
Microbiome
By applying HTP workflows, N-glycan interactions with microbial enzymes can be efficiently characterized, guiding the design of prebiotics and microbiota-targeted interventions
Biomarkers & Diagnostics
HTP N-glycan profiling (sera, plasma) provides scalable, sensitive detection of disease-associated glycan signatures, improving early diagnosis, patient stratification, and therapeutic monitoring
Biopharmaceutical Development
Process optimization and early-stage biopharmaceutical development, and batch-to-batch comparison
Other fields
- Veterinary medicine: veterinary medicine by validating glycan biomarkers and vaccines across large animal cohorts
- Plants and Agriculture: HTP N-glycan workflows accelerate discovery in plants by mapping stress-related glycosylation
- Nutraceuticals and cosmetics: this service helps identify rapidly by screening bioactive glycans
- Human Milk Oligosaccharides (HMOs): by identifying functional glycan mimetics for infant nutrition
Related publications
A selection of our most relevant scientific publications that form our HTP Profiling of N-glycans
De Leoz MLA, Duewer DL, Fung A, Liu L, Yau HK, Potter O, et al. NIST interlaboratory study on glycosylation analysis of monoclonal antibodies: Comparison of results from diverse analytical methods. Mol Cell Proteomics. 2020;19(1):11–30.
Etxebarria J, Reichardt N-C. Methods for the absolute quantification of N-glycan biomarkers. Biochim Biophys Acta. 2016;1860(8):1676–87.
