Heparin sulfate (HS) can interact with proteins and other molecules that affect many biological processes. At CD BioGlyco, researchers have developed microarrays of multiple HS structures to help clients identify specific binding interactions between various HP and their potential protein partners.

Introduction

HS is a sulfated unbranched polysaccharide found on the cell surface and in the extracellular matrix that regulates a wide range of physiological and pathophysiological processes. Variations in HS sulfation and epimerization provide great structural diversity that is thought to underlie protein binding and regulatory properties. Due to the ubiquity and multifunctionality of HS, the study of the binding roles of such glycans is important for understanding how to combat related diseases.

The specific binding motifs for HS interactions have not been extensively characterized. Glycan microarray technology has been developed to efficiently probe the interactions between glycans and their ligands while requiring relatively small amounts of samples. In addition, advances in the chemoenzymatic synthesis of HS have enabled the production of specific HS structures, providing the basis for the development of microarrays of multiple chemoenzymatically synthesized HS structures to explore the interactions of various types of HS.

Fig.1 Workflow for constructing a glycosaminoglycan microarray containing HS. (Watanabe, et al., 2021)

Our Services

CD BioGlyco is dedicated to developing technology for combining chemoenzymatic synthesis with microarray techniques to determine the specific HS sequences required for binding to proteins. We developed a methodology to prepare large numbers of differentially sulfated HS oligosaccharides via a modular approach, providing an unprecedented library of multiple HS oligosaccharides for the development of microarrays. HS microarrays are used to detect the ligand requirements of many HS-binding proteins and the data allow the selection of compounds that interfere with biological processes such as growth factor-induced cell proliferation and explore the role of changes in cell surface HS composition in regulating protein function.

Workflow

We provide a comprehensive solution from sample processing to data analysis, aiming to deliver accurate and reliable HS acid binding analysis results.

• HS microarray construction: Our microarray contains multiple carefully selected, structurally defined HS. HS is synthesized and purified using chemical and enzymatic methods, then spotted onto the carrier surface.
• Sample processing: Our HS microarray is suitable for analyzing biological molecules such as viral proteins, antibodies, growth factors, bacterial adhesins, and other biological samples. Depending on the type of sample, we perform appropriate processing and labeling to ensure sensitivity in subsequent detection.
• Incubation: Labeled samples are incubated with the HS microarray under optimized conditions to achieve specific binding. Incubation conditions (temperature, time, buffer, ionic strength) are optimized based on the characteristics of the target molecule and its binding mechanism. Unbound molecules are then removed through washing. We use state-of-the-art microarray scanners to capture high-resolution and high-sensitivity fluorescence intensity images of the binding signals.
• Data analysis: We standardize the raw fluorescence intensity data, analyze the binding strength of the target molecules with each HS oligosaccharide, and enable multi-sample comparisons.

Applications

• HS microarrays are a powerful tool for systematically studying complex glycan-protein interactions, enabling a comprehensive assessment of the binding specificity and affinity of specific proteins toward HS oligosaccharides.
• The complex and variable structure of HS plays a role in various biological functions, such as cellular signaling and molecular recognition. Microarray analysis facilitates high-throughput screening to identify and decode the correspondence between HS structures and biological functions, thereby supporting research into disease mechanisms and other areas.
• By utilizing HS microarrays as a high-throughput screening platform, small molecules, peptides, and other compounds that can specifically bind to HS, mimic HS functions, or effectively inhibit HS-protein interactions can be identified, thereby regulating HS-mediated biological processes and providing candidate molecules for new drug development.

Advantages

• Our HS oligosaccharide library is rich in variety, enabling detailed structural analysis of HS-protein interactions through microarray analysis.
• Our optimized microarray detection system ensures high sensitivity and binding specificity while minimizing non-specific background.
• The synthesis of HS oligosaccharides and microarray preparation processes are highly standardized, combined with strict quality control and experimental procedures, ensuring reproducibility of experimental results both between and within batches.

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Frequently Asked Questions

CD BioGlyco has expanded microarray technology in recent years to provide a dedicated research platform for characterizing HS interactions. We have extensive experience in microarray services and we are confident to be your partner in the field of glycobiology. If you are interested in our services, please contact us for more information.

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