Photoimmobilization of unmodified carbohydrates on activated surface.
Herein, we demonstrate a simple, versatile and efficient method for immobilization of unmodified carbohydrates onto a solid surface. The method employs a photoreactive-cellulose membrane which is prepared by the introduction of a photoreactive group to a cellulose membrane through 1-fluoro-2-nitro-4-azidobenzene (FNAB). Upon exposure to UV light photo-reactive azido group of the activated cellulose membrane transforms itself into highly reactive nitrene which covalently binds to underivatized carbohydrates through an insertion reaction. Maximum immobilization of carbohydrate is achieved at the UV exposure time of 60min and carbohydrate concentration of 100microg/disk. The immobilized carbohydrate is assayed by affinity binding of biotin-labeled lectins. The amount of Con A required for detecting immobilized carbohydrate ranges from 4-10microg/ml. The binding affinities of the lectin to the immobilized carbohydrates are analyzed by calculating their IC(50) values. Overall, the present work demonstrates an efficient immobilization of underivatized carbohydrate onto cellulose surface and has the potential to be applied to other surfaces.
Sharma
Pragya
P
Institute of Genomics and Integrative Biology (CSIR), Mall Road, Delhi 110 007, India.
Basir
Seemi Farhat
SF
Nahar
Pradip
P
D000117:Acetylglucosamine / Q000737:chemistry; D001386:Azides / Q000737:chemistry* / Q000528:radiation effects; D002241:Carbohydrates / Q000737:chemistry* / Q000528:radiation effects; D002482:Cellulose / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods; D003208:Concanavalin A / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D054794:Glycomics / Q000379:methods; D020128:Inhibitory Concentration 50; D008358:Mannose / Q000737:chemistry; D008567:Membranes, Artificial; D009578:Nitrobenzenes / Q000737:chemistry* / Q000528:radiation effects; D055668:Photochemical Processes; D013499:Surface Properties; D014466:Ultraviolet Rays
C492759:1-fluoro-2-nitro-4-azidobenzene; D001386:Azides; D002241:Carbohydrates; D008567:Membranes, Artificial; D009578:Nitrobenzenes; D003208:Concanavalin A; D002482:Cellulose; D005947:Glucose; D008358:Mannose; D000117:Acetylglucosamine