Glycosaminoglycan (GAG) Microarray Assay
Unraveling Complexity, One Glycosaminoglycan at a Time
Glycosaminoglycans (GAGs) are sulfated glycans in the extracellular matrix and on the cell surface that are critical for the regulation of various signaling proteins and whose function is essential in many pathophysiological systems. Therefore, GAGs can serve as useful molecular tools for researching GAG-protein interactions. At CD BioGlyco, we have constructed a variety of specific Glycan Microarrays based on the development of a comprehensive Glycan Display Platform. With our expertise in Glycan Microarray technology, we offer custom GAG microarray assays.
The GAG microarray and analysis services based on the GAG microarray we provided are as follows.
- GAG microarray construction
- First, we prepare the GAG-protein couplers by providing either a chemoenzymatic pathway or modular assembly based on pre-synthesized building blocks.
- Then, we dissolve the GAG-protein couplers in a spotting solution. At the same time, the insoluble material is removed by filtration.
- Next, we covalently immobilized GAG on epoxy-activated microarray slides using a non-contact printing technique.
- Finally, the glass slides are incubated. Subsequently, excess unimmobilized material is washed away with probe buffer and closed with buffer.
- GAG microarray assay
We fluorescently label proteins coupled to GAGs. The specific binding of proteins to GAGs is detected using a fluorescence scanner. Similarly, we utilize GAG microarrays to screen for GAGs recognized by the binding proteins. In this process, we provide various binding analysis techniques to analyze the binding ability of the binding proteins to different GAGs. Such as surface plasmon resonance (SPR), pull-down analysis, flow cytometry, and so on.
Fig.1 GAG microarray assay services. (CD BioGlyco)
Publication
Technology: Fluorescence detection
Journal: FEBS Letters
IF: 3.5
Published: 2021
Results: HS is a sulfated GAG, an essential host attachment factor that promotes SARS-CoV-2 infection. In this thesis, the authors have developed fluorescence detection-based GAG microarrays for high-sensitivity screening of proteins for GAG-binding specificity and apply them to the analysis of SARS-CoV-2 spiny (S) proteins. Out of 20 different GAGs, S proteins bind not only to heparin (HEP)/HS but also to chondroitin sulfate E (CSE) in a concentration-dependent manner. Next, the authors analyzed the specificity of each subunit of S proteins. While the S1 subunit displayed exclusive binding to HEP, the S2 subunit was also bound to CSE and HP/HS. CSE may act as a surrogate attachment factor for HS in SARS-CoV-2 infection.
Applications
- GAG microarrays can be used to detect proteins or glycoproteins that bind specifically to GAGs.
- GAG microarrays can be used for high-throughput screening of GAG-mediated protein-cell interactions.
- GAG microarrays can be used to research GAG-protein interactions, leading to the preparation of GAGs that are valuable for physical and pharmaceutical research.
Advantages
- We construct GAG microarrays with high sensitivity and high throughput screening capabilities, and only a trace amount of sample is needed to complete the entire assay.
- With a professional R&D team, we provide custom glycan microarray assays according to clients' needs.
- In addition to glycan microarray assays, we also provide Microarray-based Glycoproteomics Services.
CD BioGlyco is dedicated to research in glycobiology and provides clients with advanced glycan microarray detection technologies. Currently, we have constructed a state-of-the-art glycan display platform, which covers a wide range of glycan microarrays and glycan display array-related services. Please feel free to contact us if you are interested in our services.
References
- Watanabe, T.; et al. A glycosaminoglycan microarray identifies the binding of SARS-CoV-2 spike protein to chondroitin sulfate E. FEBS Letters. 2021, 595(18): 2341-2349.
- Pomin, V.H.; Wang, X. Synthetic oligosaccharide libraries and microarray technology: a powerful combination for the success of current glycosaminoglycan interactomics. ChemMedChem. 2018, 13(7): 648-661.
