Trityl-derivatized carbohydrates immobilized on a polystyrene microplate.
Carbohydrate biosensors, including carbohydrate arrays, are attracting increased attention for the comprehensive and high-throughput investigation of protein-carbohydrate interactions. Here, we describe an effective approach to fabricating a robust microplate-based carbohydrate array capable of probing protein binding and screening for inhibitors in a high-throughout manner. This approach involves the derivatization of carbohydrates with a trityl group through an alkyl linker and the immobilization of the trityl-derivatized carbohydrates (mannose and maltose) onto microplates noncovalently to construct carbohydrate arrays. The trityl carbohydrate derivative has very good immobilization efficiency for polystyrene microplates and strong resistance to aqueous washing. The carbohydrate arrays can probe the interactions with the lectin Concanavalin A and screen this protein for the well-known inhibitors methyl alpha-D-mannopyranoside and methyl alpha-D-glucopyranoside in a high-throughput manner. The method described in this paper represents a convenient way of fabricating robust noncovalent carbohydrate arrays on microplates and offers a convenient platform for high-throughput drug screening.
Zou
Lan
L
Department of Applied Biology and Chemical Technology, Central Laboratory of the Institute of Molecular Technology for Drug Discovery and Synthesis, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, PR China.
Pang
Hei-Leung
HL
Chan
Pak-Ho
PH
Huang
Zhi-Shu
ZS
Gu
Lian-Quan
LQ
Wong
Kwok-Yin
KY
D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000138:chemical synthesis / Q000737:chemistry*; D003208:Concanavalin A / Q000737:chemistry; D004187:Disaccharides / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry; D007700:Kinetics; D037102:Lectins / Q000737:chemistry; D008958:Models, Molecular; D011137:Polystyrenes; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D014320:Triphenylmethyl Compounds / Q000737:chemistry*
D002241:Carbohydrates; D004187:Disaccharides; D005452:Fluoresceins; D006023:Glycoproteins; D037102:Lectins; D011137:Polystyrenes; D011506:Proteins; D014320:Triphenylmethyl Compounds; D003208:Concanavalin A