Carbohydrate microarray for the detection of glycan-protein interactions using metal-enhanced fluorescence.
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.
†Physique de la Matière Condensée, Ecole Polytechnique-CNRS, 91128 Palaiseau, France.
Jie
J
Yang
†Physique de la Matière Condensée, Ecole Polytechnique-CNRS, 91128 Palaiseau, France.
Anne
A
Moraillon
‡Laboratoire de Glycochimie des Antimicrobiens et des Agroressources (LG2A), (FRE 3517-CNRS), Université de Picardie Jules Verne, 33 Rue St Leu, 80039 Amiens, France.
Aloysius
A
Siriwardena
§Institut d'Electronique, de Microélectronique et de Nanotechnologie (IEMN, CNRS-8520), Université Lille 1, Cité Scientifique, Avenue Poincaré B.P. 60069, 59652 Villeneuve d'Ascq, France.
Rabah
R
Boukherroub
†Physique de la Matière Condensée, Ecole Polytechnique-CNRS, 91128 Palaiseau, France.
François
F
Ozanam
†Physique de la Matière Condensée, Ecole Polytechnique-CNRS, 91128 Palaiseau, France.
Anne Chantal
AC
Gouget-Laemmel
§Institut d'Electronique, de Microélectronique et de Nanotechnologie (IEMN, CNRS-8520), Université Lille 1, Cité Scientifique, Avenue Poincaré B.P. 60069, 59652 Villeneuve d'Ascq, France.
Sabine
S
Szunerits
D001386:Azides / Q000737:chemistry; D057930:Click Chemistry; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D037102:Lectins / Q000032:analysis / Q000378:metabolism*; D046228:Microarray Analysis / Q000295:instrumentation*; D049329:Nanostructures / Q000737:chemistry*; D061848:Optical Imaging / Q000295:instrumentation*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*
D001386:Azides; D005456:Fluorescent Dyes; D037102:Lectins; D011134:Polysaccharides; D006046:Gold