Hydrogel glycan microarrays

• Published in: Analytical biochemistry 2005-12, Vol.347 (1), p.94-105
• Main Authors: Dyukova, V.I., Dementieva, E.I., Zubtsov, D.A., Galanina, O.E., Bovin, N.V., Rubina, A.Yu
• Format: Article
• Language: English
• Subjects: ABO Blood-Group System - analysis; Antibodies; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; Carbohydrates; Combined glyco/protein chip; Gel microchips; Glycan microarrays; Humans; Hydrogel glycochips; Hydrogel, Polyethylene Glycol Dimethacrylate - chemistry; Lectins; Molecular Probes - chemistry; Polysaccharides - chemistry; Protein Array Analysis; Ricin; Ricin - chemistry; Trisaccharides - chemistry
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2005.09.009

The technology of hydrogel microchips manufacturing, which was developed previously for covalent immobilization of DNA and proteins, was applied for the preparation of glycochips and combined glyco/protein chips. Microchips consist of hydrogel drops separated with hydrophobic surface. Spacered amino-saccharides and polyacrylamide glycoconjugates were used for immobilization. Gel elements were ∼1 nl in volume (150 μm in diameter and 25 μm in height), and the amount of covalently immobilized saccharide in the glycoarray was 0.4–1.7 pmol per gel element. Hydrogel glycan microchips were used for quantitative assay of antibodies against blood group antigens and assay of lectins with fluorescent detection. In all cases, only specific interaction with chip-immobilized saccharides was observed, whereas the background signal was very low. The detection limit of on-chip assays was comparable to that of the standard 96-well plate assays. Mixing of reaction solution allowed us to decrease the duration of the assays significantly: 2–3 h for incubation and development steps and 10 min for washing. A method for determination of association constants for binding of compounds with chip-immobilized ligands from the kinetics of their binding is proposed. Combined microchips containing different types of biomolecules can be designed and used for simultaneous detection of different compounds.