100 N-Glycan Microarray

Introduction

Glycans are essential in numerous physiological and pathological processes, including cell signaling, immune modulation, host-pathogen interactions, and cancer progression. Among them, N-linked glycans represent a structurally diverse and biologically critical class of oligosaccharides covalently attached to asparagine residues within glycoproteins. Yet, understanding their interactions remains challenging due to their heterogeneity and complex biosynthesis. To overcome this, Creative Biolabs offers a 100 N-glycan microarray platform, a robust and reproducible analytical solution to systematically evaluate the binding properties of glycan-interacting molecules. This platform enables comprehensive profiling of glycan-binding proteins (GBPs), antibodies, viral particles, and cells against a standardized panel of 100 well-characterized N-glycans.

What Is the 100 N-Glycan Microarray?

The 100 N-glycan microarray is a pre-printed slide containing 100 structurally verified N-glycans, selected to represent major mammalian glycosylation motifs. These include high-mannose, hybrid, and complex-type N-glycans with varied terminal residues such as sialic acids (Neu5Ac/Neu5Gc), galactose, fucose, and bisecting GlcNAc. Each glycan is covalently linked to a glass surface via a spacer arm, ensuring optimal spatial orientation and accessibility for binding assays. Slides are compatible with fluorescence-based detection and surface-sensitive imaging systems.

Why Analyze N-Glycan Interactions?

N-Glycans modulate protein folding, trafficking, and clearance. Importantly, specific N-glycan motifs act as ligands for various lectins, pathogens, or immune receptors. For instance:

• DC-SIGN, Siglecs, and galectins bind distinct N-glycan structures to regulate inflammation or tolerance.
• Tumor-specific glycan signatures serve as diagnostic and therapeutic targets.
• Viruses such as influenza, HIV, and coronaviruses exploit host N-glycans for cell entry.
• Glycoengineering of therapeutic antibodies requires precise knowledge of N-glycan-mediated effector functions.

Step by Step Workflow for N-glycan Microarray

Fig.1 Standardized workflow for 100 N-glycan microarray platform.

1. Array Fabrication: Covalent attachment of 100 purified N-glycans to a chemically activated glass surface.
2. Sample Incubation: Application of GBPs, antibodies, viruses, or live cells labeled with a fluorescent tag or followed by labeled secondary antibodies.
3. Washing & Detection: Removal of unbound material and acquisition of signal via laser scanner or microscope.
4. Data Analysis: Generation of heatmaps, glycan specificity profiles, and statistical clustering using proprietary software.

Data Interpretation & Reporting

Results are provided in a structured, publication-ready format. Our deliverables include:

• Fluorescent signal heatmaps
• Quantitative glycan-binding intensity data
• Glycan specificity clustering (PCA or hierarchical)
• Comparative plots for group analysis

Advantages of the Creative Biolabs Platform

• Structural Fidelity: All glycans are chemically or enzymatically synthesized to ensure high purity and accurate linkage definition, confirmed by LC-MS/MS.
• Customizable Microarrays: Beyond the standard 100-N-glycan panel, we offer custom spotting with additional human or pathogen-associated glycans, glycopeptides, or lipid-linked glycoconjugates.
• Low Sample Requirement: Only ~5–20 µg of test material is needed, making this suitable for rare antibodies or expensive recombinant proteins.
• Cross-Species Applications: Validated for use with human, mouse, rat, and non-human primate samples. Compatible with academic research.

Applications in Biomedical Research

Fig.2 The application of N-glycan microarray in research.

Lectin Specificity Profiling

Lectins are glycan-binding proteins essential for immune recognition, cell adhesion, and intracellular trafficking. The microarray can reveal subtle preferences of lectins for specific glycan epitopes, including fucosylation and sialylation variants.

Antibody Glycan Specificity

In vaccine development and infectious disease studies, antibody-glycan interactions are crucial. For example, anti-glycan antibodies targeting tumor-associated carbohydrate antigens (TACAs) can be validated for glycoepitope specificity using the array.

Pathogen-Glycan Interactions

Viral envelope proteins (e.g., hemagglutinin, spike glycoproteins) and bacterial adhesins often target host N-glycans. Screening these proteins on the 100 N-glycan array helps identify glycan-based entry mechanisms and therapeutic targets.

Cancer Biomarker Discovery

Aberrant N-glycosylation is a hallmark of many cancers. Profiling serum or tumor-derived proteins for altered glycan-binding profiles provides insights into disease progression and potential diagnostic markers.

The 100 N-glycan microarray platform from Creative Biolabs offers a powerful solution for elucidating glycan-binding interactions with high sensitivity, reproducibility, and biological relevance. This technology accelerates glycomics discovery and biomarker development for basic research. Whether characterizing immune receptors, mapping therapeutic antibody specificities, or profiling pathogen tropism, our platform provides the precision and throughput that modern glycobiology demands. Contact us to get your tailored and time-saving solutions for N-glycan related research.

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FAQs

Reference:

1. Bojar, Daniel, et al. "A useful guide to lectin binding: machine-learning directed annotation of 57 unique lectin specificities." ACS chemical biology 17.11 (2022): 2993-3012. https://doi.org/10.1021/acschembio.1c00689

Supports

• N-Linked Glycan