The hydroxyl-modified surfaces on glass support for fabrication of carbohydrate microarrays.
Glycan-protein interactions play important biological roles in biological processes. But there is a lack of simple high-throughput methods to elucidate recognition events between carbohydrates and protein. Although, there have been a number of glycan arrays developed in recent years utilizing different strategies and for different purposes, the method presented in this paper, a direct covalent immobilization of sugars to hydroxyl-modified glass surface, can be a very useful general method. Here, two strategies have been developed for the production of carbohydrate microarrays by the underivatized sugars that efficiently immobilized on hydroxyl-functionalized glass surface by formation of glycosidic bond with the hemiacetal group at the reducing end of the suitable carbohydrates via condensation. Firstly, untreated glass slides were amino- and epoxy-silanized, respectively. Then, they were further hydroxyl functionalized with different surface chemistries. The carbohydrate microarrays were fabricated on hydroxyl-functionalized glass by robotic arrayer. Additionally, experiments for the characterization of carbohydrates-protein interaction were performed to compare these strategies. Overall best results in terms of conveniency and sensitivity were obtained with hydroxyl-functionalization on epoxysilanized surfaces. The hydroxyl-functionalized slide was used to detect the amount of mannose immobilized on the glass surface. The limit of detection of the fabricated mannose microarray was 100 nM.
Nan
Gang
G
College of Life Sciences, Northwest University, 710069 Xi'an, P.R. China.
Yan
Hua
H
Yang
Ganglong
G
Jian
Qiang
Q
Chen
Chao
C
Li
Zheng
Z
D002241:Carbohydrates / Q000138:chemical synthesis*; D005898:Glass / Q000737:chemistry*; D006878:Hydroxides / Q000138:chemical synthesis*; D046228:Microarray Analysis / Q000379:methods* / Q000639:trends; D012371:Robotics / Q000639:trends; D013499:Surface Properties
D002241:Carbohydrates; D006878:Hydroxides; C031356:hydroxide ion