Carbohydrate Microarray for the Detection of Glycan–Protein Interactions Using Metal-Enhanced Fluorescence

Abstract

Carbohydrate arrays are potentially one of the most attractive tools to study carbohydrate-based interactions. This paper describes a new analytical platform that exploits metal-enhanced fluorescence for the sensitive and selective screening of carbohydrate-lectin interactions. The chip consists of a glass slide covered with gold nanostructures, postcoated with a thin layer of amorphous silicon-carbon alloy (a-Si0.8C0.2:H). An immobilization strategy based on the formation of a covalent bond between propargyl-terminated glycans and surface-linked azide groups was used to attach various glycans at varying surface densities onto the interface and to fabricate a carbohydrate array via efficient local "click" chemistry strategy. The specific association of the new interface with fluorescently labeled lectins was assessed by fluorescence imaging and an excellent selectivity to specific proteins was achieved. Optimization of the surface architecture and the plasmonic transducer resulted in an enhancement of the fluorescence intensity by 1 order of magnitude, when compared to the corresponding substrate devoid of gold nanostructures. The limit of detection (LOD) of such microarrays is in the picomolar range, making it a promising system for development in pharmaceutical or biomedical applications.

Vitronectin (Vn) glycosylation patterned by lectin affinity assays—A potent glycoproteomic tool to discriminate plasma Vn from cancer ascites Vn

“…Thus, the commercial lectin array we used appears suitable for high‐throughput screening and profiling of glycosylation and glycoproteins. Accumulating propositions based on microarray technology and lectin affinity are gaining popularity in glycomics and glycoproteomics . Thanks to their diversity and specificity of glycan recognition, lectins can distinguish between linkages or other fine nuances of oligosaccharide, information not easily accessible using mass spectrometry or chromatography methods .…”

Rapid and sensitive identification of uropathogenic Escherichia coli using a surface-enhanced-Raman-scattering-based biochip

“…[44][45] The integration of undecylenic acid functions onto a-Si:H is based on the photochemical hydrosilylation reaction, [46] followed by the amidation of a bifunctional oligo(ethylene glycol) OEG linker, HOOC-OEG 12 -NH 2 in a two-step process using the wellknown EDC/NHS coupling activation. [47][48] The presence of at least 8 OEG units proved to be highly efficient for limiting non-specific surface interactions with proteins like lectins. To demonstrate the reliability of this surface functionalization scheme, FTIR-ATR measurements were performed after each reaction step on a 20 nm-thick a-Si:H film deposited on a silicon ATR prism.…”

Analytical strategies used to identify the readers of histone modifications: A review

“…The microarray format is a representative of high-throughput techniques, in which the probes are immobilized on a suitable supporting material and exposed to samples containing potential binders. After extensive washing, the retained specific binders can be further analyzed and identified [24,63,64] .…”