Immunoglobulins are glycoproteins produced by B cells that play key roles in the immune response. They are classified into five major types: IgG, IgA, IgM, IgE, and IgD.
Adapted from Vattepu R et al. (2022) Sialylation as an Important Regulator of Antibody Function. Front. Immunol. 13:818736.
All immunoglobulins are glycosylated, and their attached glycans are essential for maintaining structural integrity, stability, and modulating immune functions.
Asparia provides a specialized immunoglobulin profiling service using advanced LC-MS and MALDI-TOF platforms, enabling in-depth characterization of immunoglobulins.
MALDI-TOF profiling of immunoglobulins released N and O-glycans
This technique, applied to Ig, enables rapid screening and fingerprinting of immunoglobulin glycoforms.
Despite being rare in therapeutic antibodies, O-glycosylation can be detected by MALDI-TOF after chemical release and permethylation. Asparia is one of the rare specialists of O-glycan analysis on the market.
We offer:
- Serum IgG N-glycan Profiling by MALDI-TOF MS + CarboQuant Technology for absolute quantification
- Recombinant therapeutic IgG Fc-glycan profiling by MALDI-TOF MS and CarboQuant Technology
UHPLC-FLD-MS profiling of immunoglobulins released N-glycans
UHPLC-MS offers significantly better quantitative accuracy and analytical resolution compared to MALDI, thanks to its combination of hydrophilic interaction chromatography (HILIC) and high-resolution mass spectrometry. This setup enables effective separation of glycan isoforms and precise quantification based on peak areas. To enhance MS signal sensitivity and facilitate fluorescence detection when needed, N-glycans are commonly labeled with tags such as procainamide or Rapifluor.
- Serum IgG-N-glycan profiling for biomarker studies
UHPLC-MS profiling of N-glycans from human plasma IgG.
- Fc/Fab fragments glycosylation analysis: IgG molecules are enzymatically cleaved to separate Fc and Fab, which are then purified. The N-glycans from each fragment can be specifically profiled to assess glycosylation patterns unique to each region. Fc glycosylation is critical, as it can influence antibody stability, effector functions (e.g., ADCC and CDC activity), and overall therapeutic efficacy. In contrast, Fab glycosylation may impact antigen binding.
How we work for immunoglobulins N-Glycan Profiling
Our workflow ensures a clear, straightforward, and transparent process for sample glycoanalysis
Sample
Send us your sample (purified Ig, serum, plasma) and we will apply our Ig-specific and improved 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.
Related publications
A selection of our most relevant scientific publications that form our Immunoglobulins N-Glycan Profiling analysis
Álvarez J, Parody N, Calzada D, Aranda T, Renshaw A, Serna S, et al. Correlation between N-glycan GnGnXF3 and the allergic immune response against Juniperus ashei pollen. Allergy. 2025.
Alvarez J, Aranda T, Ruiz R, Aramendia L, Reichardt N-C, Carnes J. N-glycans other than MUXF3 are targets for specific IgE antibodies against Juniperus ashei pollen allergens. J Allergy Clin Immunol. 2023;151(2):AB212.
Huettner I, Krumm SA, Serna S, Brzezicka K, Monaco S, Walpole S, et al. Cross-reactivity of glycan-reactive HIV-1 broadly neutralizing antibodies with parasite glycans. Cell Rep. 2022;38(13):110611.
Robakiewicz S, Bridot C, Serna S, Gimeno A, Echeverria B, Delgado S, et al. Minimal epitope for Mannitou IgM on paucimannose-carrying glycoproteins. Glycobiology. 2021;31(8):1005–17.
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.
Vattepu, R.; Sneed, S. L.; Anthony, R. M. Sialylation as an Important Regulator of Antibody Function. Front. Immunol. 2022, 13.
Purposes and applications for N-Glycan Profiling
N-glycans of a purified immunoglobulin serve several purposes and applications:
Oncology
Reveals tumor-associated changes in antibody glycosylation that affect immune surveillance, supporting biomarker discovery and glycoengineered cancer therapeutics
Biomarkers & Diagnostics
Disease-specific immunoglobulin N-glycan signatures serve as robust biomarkers for early diagnosis, patient stratification, and longitudinal monitoring of therapy
Infectious Diseases
In infectious diseases, complex and altered N-glycans are exploited by pathogens for cell entry; profiling these interactions informs vaccine design and antiviral glycomimetics
Immune and Inflammatory Diseases
Uncovers shifts in Fc glycosylation linked to inflammation and autoimmunity (generated during allergies and inflammatory processes) , enabling sensitive diagnostics and guiding therapeutic antibody design
Microbiome
Alterations in host immunoglobulin N-glycans reflect interactions with the microbiota, providing insights into mucosal immunity and supporting microbiota-targeted interventions
Other Fields
- Process optimization and early-stage biopharmaceutical development, batch-to-batch comparison in the biosimilar field
- Veterinary medicine: it aids in diagnosing animal diseases and improving vaccine efficacy
- Nutraceuticals and Cosmetics: it supports the validation of immune-modulating glycans
- Human Milk Oligosaccharides (HMOs): inform the design of glycan mimetics that enhance infant immunity
