Issue 9, 2012

Surface chemistry and linker effects on lectin–carbohydrate recognition for glycan microarrays

Abstract

Glycan microarrays are an increasingly utilised tool for analysis of proteincarbohydrate interactions and a variety of glycan-containing molecules and slide chemistries have been used to array carbohydrates on microarray surfaces. Slide surface chemistry can have significant impact on the ligand presentation, background noise, spot size and morphology and reproducibility of the arrayed molecules, which in turn impacts upon lectin–carbohydrate recognition. The linker used to attach the carbohydrate to the molecular scaffold is another variable in ligand presentation. To evaluate these effects, three different microarray surface chemistries were arrayed with the same mono- and di-saccharide neoglycoconjugates and natural glycoproteins and incubated with four well-characterised plant lectins. Analogues of three monosaccharide neoglycoconjugates, with two common linkers each, were included in the test group to evaluate the linker effect on lectin recognition. Based on lowest background noise, expected lectin–ligand interaction, good spot morphology and best reproducibility, the three-dimensional hydrogel slide surface proved most suitable for lectin interrogation of carbohydrate ligands, and the more flexible phenylisothiocyanate linker afforded greater recognition of the carbohydrates by the relevant lectins.

Graphical abstract: Surface chemistry and linker effects on lectin–carbohydrate recognition for glycan microarrays

Supplementary files

Article information

Article type
Paper
Submitted
24 May 2012
Accepted
18 Jul 2012
First published
18 Jul 2012

Anal. Methods, 2012,4, 2721-2728

Surface chemistry and linker effects on lectin–carbohydrate recognition for glycan microarrays

M. Kilcoyne, J. Q. Gerlach, M. Kane and L. Joshi, Anal. Methods, 2012, 4, 2721 DOI: 10.1039/C2AY25532D

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