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Glycopeptide Microarray

Glycopeptide Microarray

Antibodies against glycoproteins or glycopeptides generally recognize the peptide backbone as a portion of the binding epitope. Therefore, CD BioGlyco has developed a highly sensitive glycopeptide microarray platform to help our customers screen antibodies against specific glycopeptide antigens. We look forward to providing you with professional and rapid solutions in the research related to glycobiology microarray.

Background

Abnormal post-translational variants of proteins may induce autoantibodies, and the change of O-linked glycosylation is one of the most significant cancer-related post-translational modifications. Short aberrant O-glycans on proteins may introduce new glycopeptide epitopes that can trigger autoantibodies. Therefore autoantibodies against cancer antigens are expected to serve as biomarkers for early cancer detection.

Currently, a new technology of glycopeptide microarray is being developed to screen for antibodies against different glycopeptide antigens and to detect glycopeptide-specific autoantibodies. Glycopeptide microarrays offer the advantages of parallelism, automation and rapidity. The parallel data detected has less error and higher precision. The emergence of this technology has played an important role in the advancement of many disciplines such as biology, genetics, clinical laboratory medicine and oncology.

(CD BioGlyco)Fig.1 Microarray platform for detection of MUC1 O-glycopeptide-specific antibodies. (Wandall, 2010)

Our Services

CD BioGlyco has developed glycopeptide microarray technology to help our customers map antibody epitopes and identify key residues for glycopeptide binding. Services we provide include but are not limited to:

  • We pre-synthesize glycopeptides of various structures and immobilize them on microarray slides. In several circumstances, glycopeptides are modified by specific enzymes (e.g. glycosyltransferases) in solution or in situ on the microarray slide. Constructed glycopeptide microarrays are incubated with interaction partners and then detected by fluorescence or other methods for binding events.
  • We use a chemoenzymatic approach to produce cancer-associated O-glycopeptide libraries in combination with a microarray platform for high-throughput detection of autoantibodies to the O-glycopeptidome. This technology allows sensitive detection of cancer-related autoantibodies to different O-glycopeptide epitopes.

Special Features

Our glycopeptide microarrays offer multiple features and provide an unparalleled high-resolution platform for precise analysis of key interactions based on various glycopeptide sequences.

  • Site-specific analysis: Glycopeptide microarray assay helps identify the impact of changes in specific glycosylation sites on interactions and recognition. For example, it helps understand how glycopeptide motifs influence their interactions with immune cells to explore disease-related changes.
  • Well-defined glycan types: We utilize glycopeptides synthesized chemically or enzymatically, with both glycan structures and peptide sequences known, ensuring unparalleled specificity and reproducibility.
  • High-throughput analysis: Microarrays are suitable for high-throughput analysis to accelerate the discovery of novel interactions and biomarkers.

When biological samples (such as serum, cell lysates, or single cells) are applied to microarrays, glycoprotein-binding proteins (GBPs), antibodies, or other interaction partners selectively bind to corresponding glycopeptide targets. Subsequently, high-sensitivity detection is used to reveal complex mechanisms.

Workflow

Our glycopeptide microarray service is carefully designed to provide reliable, high-quality data.

  • Glycopeptide synthesis and chip preparation: We use chemical or enzymatic methods to precisely synthesize glycopeptides with diverse structures. These glycopeptides are verified using mass spectrometry (MS), nuclear magnetic resonance (NMR), and other techniques. These highly purified glycopeptides are then carefully spotted onto specially treated solid surfaces using advanced printing technology in a precise, high-density array format. Our rigorous preparation process ensures the purity, integrity, and uniform presentation of each glycopeptide on the array.
  • Sample preparation and microarray incubation: Our glycopeptide microarray analysis services are applicable to various biological samples, such as serum, plasma, and cell lysates. After receiving the samples, we perform appropriate processing, including purification and labeling. The samples are then incubated with the glycopeptide microarray under strictly controlled conditions to facilitate the occurrence of specific binding reactions.
  • Signal detection and data acquisition: After incubation, non-specific binding molecules are removed through rigorous washing steps, followed by scanning of the microarray using a high-resolution, high-sensitivity microarray scanner. Advanced detection technology ensures the capture of even weak binding events, providing relevant quantitative data.
  • Data analysis and interpretation: Raw intensity data undergoes a comprehensive bioinformatics processing workflow. Our specialized analysis transforms complex data into clear, actionable insights, highlighting key biological discoveries and potential biomarkers.

Process of our 100 glycan microarray analysis. (CD BioGlyco)

Advantages

  • Wide range of applications: Our services are compatible with a variety of biological samples, including body fluids, cell lysates, plasma, and more from different sources. This broad applicability makes it useful in many research fields.
  • Minimal sample requirements: Analysis requires only a small amount of precious biological samples, making it an ideal choice for testing rare samples.
  • High sensitivity and quantitative data: Our services are suitable for detecting subtle differences in binding affinity and low-abundance interactions, providing reliable quantitative insights for specific glycan type identification.

Applications

  • Biomarker discovery: Glycopeptide microarray analysis is suitable for identifying novel disease-specific glycan type biomarkers for early detection, prognosis assessment, and monitoring of cancers, autoimmune diseases, and more.
  • Vaccine development: Glycopeptide microarray analysis is suitable for analyzing vaccine binding characteristics, providing useful information for vaccine development.
  • Basic glycobiology research: Glycopeptide microarray analysis is suitable for elucidating the binding preferences of GBPs, thereby helping to reveal how these proteins recognize and bind to targets on the cell surface or in the extracellular matrix.

Publication Data

Technology: Glycopeptide microarray

Journal: Journal of the American Chemical Society

IF: 14.4

Published: 2023

Results: Macrophage galactose-type lectin (MGL) is a popular target for cancer vaccines. In this study, the researchers used solid-phase peptide synthesis to make a candidate vaccine that targets MGL on antigen-presenting cells, which has a high-affinity sugar cluster based on a tetraphenylethylene scaffold. The study found that immunization with this newly designed vaccine construct carrying GalNAc glycan clusters induced higher titers of anti-Tn-MUC1 antibodies. The antibodies obtained could bind to tumor-associated glycan structures on MUC1 and MUC1-positive breast cancer cells. Researchers assessed antibody selectivity using a glycopeptide microarray binding assay, which showed that the glycan clusters do not affect the recognition pattern of the antibodies. This study provides support for the design and development of vaccines targeting MGL.

Fig.1 Glycopeptide microarray used to analyze binding specificity.Fig. 1 Antibody binding specificity to the mucin core and glycopeptides produced by vaccines 15 and 16. (Gabba, et al., 2023)

Frequently Asked Questions

With a large and well-equipped platform, CD BioGlyco provides high-quality glycopeptide microarray services to clients around the world, and we have comprehensive capabilities to meet your specific needs on glycopeptide microarray analysis. If you need our services, please contact us for more details and technical support.

Associated Services

Reference

  1. Gabba, A.; et al. MUC1 glycopeptide vaccine modified with a GalNAc glycocluster targets the macrophage galactose C-type lectin on dendritic cells to elicit an improved humoral response. Journal of the American Chemical Society. 2023, 145(24): 13027-13037. (Open Access)
This service is for Research Use Only, not intended for any clinical use.
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CD BioGlyco is a world-class biotechnology company with offices in many countries. Our products and services provide a viable option to what is otherwise available.

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