Everything You Want to Know About Glyco-Microarray Technology
Microarray technology is a collection of many microscopic chemical reaction regions, which was developed in 2002 for glycobiology research. It involves immobilizing thousands of oligosaccharide sequences in the form of dot arrays on specific substrates and analyzing the specific interactions between sugar molecules and other biomolecules by high-throughput scanning techniques to study the functions and mechanisms of action of glycans in living organisms.
The key to the preparation of sugar microarrays is the appropriate method of immobilizing sugar molecules on solid-phase carriers, which relies heavily on the development of materials chemistry.
The preparation and analysis of glycan microarrays is similar to that of gene chips, and various automated devices used for DNA array technology can also be used for glycan microarray studies. Immobilization of glycans in glyco-microarrays is mainly based on the immobilization and covalent immobilization of sugar molecules by physical adsorption on the surface of the carrier. Most immobilization methods involve immobilizing the glycan ligands on the carrier surface to form a two-dimensional monomolecular layer.
The main steps of glycan microarray preparation include:
(1) Glycan library creation. The method is similar to the establishment of gene libraries and protein libraries. The main sources of glycans are through chemical synthesis, enzymatic synthesis and biosynthesis.
(2) Immobilization of the sample. Covalent immobilization is mainly through chemical reactions between sugar molecules and carriers to form covalent bonds, and non-covalent immobilization is mainly through adsorption between sugar molecules and solid phase carriers.
(3) Detection of labeling and data processing. The labeling methods mainly include fluorescent labeling and radiolabeling, and the detection methods mainly rely on matrix scanner as well as mass spectrometry-liquid chromatography.
Glycan microarray construction methods are key to glycan microarray technology, and certain immobilization methods that rely on non-specific adsorption between sugars and supports have been successfully used for glycobiological studies such as oligosaccharides, glycans, glycolipids, glycoproteins and glycoconjugates.
Covalently linked mono/bisaccharide microarrays
Covalently linked sugar microarrays are formed by immobilizing mono(bis)sugars by covalent linkage on a glass sheet carrier encapsulated with a monolayer of activation groups. Generally, the mono(bis)sugar molecule is first derivatized and then converted into a sugar derivative containing a cis-butylene-diimide covalent bond at the C6 position of the sugar ring. This cis-butylene-diimide coplanar group is attached to the carrier by addition or cycloaddition reactions with thiol or benzoquinone groups on the surface of the carrier.
Biskup et al. proposed a method to immobilize sugars containing aldehyde (amine) groups on amidated (formylated) glass sheets by reductive amidation or amidation. Angeloni et al. also developed a universal microarray platform based on dextran-coated glass sheets for the covalent immobilization of glycans and lectins, among others.
Non-covalent adsorption polysaccharide microarrays
The hydrophobic adsorbent sugar microarray technology involves the immobilization of polysaccharides or glycoproteins on a surface hydrophobic carrier by non-covalent adsorption. The high molecular weight and hydrophobicity of the polysaccharide determine its ability to bind to the carrier surface by non-covalent adsorption. The carrier surface is usually coated with a polymeric film to increase the surface hydrophobicity. The hydrophobic surface of the carrier is air-dried to improve the non-covalent immobilization of the polysaccharide on the carrier.
Oligosaccharide molecules cannot be microarrayed by non-covalent immobilization methods because they consist of only 2~20 monosaccharides and have low molecular weight and hydrophilic properties compared with polysaccharides. To overcome this drawback, glycolipid microarray technology can be realized by attaching a lipid molecule to the oligosaccharide molecule.
Lectin microarrays allow access to core information on glycan structures such as N2 glycosylation or O2 glycosylation, high mannose type or complex type, full or partial neuraminylation, etc. by lectin affinity techniques.
The most distinctive feature of glyco-microarray technology is the small amount of samples and the large number of samples tested at once, which can be analyzed and identified for hundreds of samples simultaneously. Glyco-microarrays can immobilize numerous glyco-ligands on a very small chip and perform multi-parameter assays on a large number of samples simultaneously under the same conditions. Thus microarrays can provide a more sensitive and appropriate analytical system than conventional methods. Sugar microarrays can directly characterize glyco-protein interactions, and the adsorption of sugars on the carrier surface can also effectively mimic the representation of these compounds on cell membranes.
As one of the leading companies in the field of histology, Creative Proteomics has many years of experience in histology research and offers many types of glycan-related microarray analysis methods to meet your various analytical needs.
· Glyco-gene Microarray Assay
· Glycan Microarray Assay
· Glycopeptide Microarray Assay