Carbohydrates and their conjugates are involved in various biological events, including viral and bacterial infection, the immune response, differentiation and development, and the progression of tumor cell metastasis. Glycan arays are anew technology that has enabled the high-sensitivity and rapid analysis carbohydrate-protein interaction and contribute to significant advances in glycomics. Glycan arays use aminute amount of materials and can be used for high-throughput profling and quantative analysis and provide information for the development of carbohydrate-based vaccines and new drug discover.
General method for creation and use of glycan microarrays [Alyssa M. McQuillan 2019]
Creative Proteomics has many years of experience analyzing defined, as well as, shotgun glycan microarrays. Our glycan arrays system provides a high throughput platform for rapid characterization of glycosylation in proteins, antibodies, hormones, and other glycoconjugates. Our staffs also can carry out assays using your glycan binding proteins (GBP) or organism(s) and assist in interpretation of data using bioinformatic software developed in house.
The ideal glycan assay is characterized by high sensitivity, quantitative analysis and high throughput analysis without labeling of the analytes. At present, the commonly used detection and analysis methods include fluorescence spectroscopy, surface plasmon resonance (SPR) technology, mass spectrometry, and isotope labeling detection methods.
Isotope labeling method (Radio-labeled) commonly used isotopes P and 3P as labels. and the signal is detected by darkroom pressure X-ray or directly with a phosphor screen imaging system. However, isotope labeling will pollute the environment and damage human health, so there are still many details that need to be paid attention to in the application.
SPR technology can directly detect the cross-linking of a glycan and a protein without fluorescent labeling, and is suitable for detecting the interaction between glycan ligands with low affinity and protein receptors. The advantage of this technology is that it can not only monitor the binding of two molecules in real time, but also It is also possible to quantitatively measure the strength of binding affinity between the two. It enables rapid determination of specific interactions between glycans and proteins and also allows analysis of enhancers or inhibitors. The disadvantage is that the limited number of samples per screening does not allow for high throughput analysis.
Mass spectrometry is also an important method for glycan microarray detection, allowing the analysis of specific structures of glycoprotein samples or glycan molecular probes without target labeling and with high sensitivity, but again not for high throughput analysis.
Labeling chemistry is widely used in sugar microarrays and is the earliest and more mature method for sugar microarray detection. Among them, fluorescence spectroscopy is the most commonly used detection method, which is safe, has high detection sensitivity, easy to operate, and allows quantitative analysis and high-throughput screening of glycoprotein interactions.