Covalent immobilization of carbohydrates on sol-gel-coated microplates.
Carbohydrate microarrays have attracted increasing attention in recent years because of their ability to monitor biologically important protein-carbohydrate interactions in a high-throughput manner. Here we have developed an effective approach to immobilizing intact carbohydrates directly on polystyrene microtiter plates coated with amine-functionalized sol-gel monolayers. Lectin binding was monitored by fluorescence spectroscopy using these covalent arrays of carbohydrates that contained six mono- and di-saccharides on the microplates. In addition, binding affinities of lectin to carbohydrates were also quantitatively analyzed by determining IC(50) values of lectin-specific antibody with these arrays. Our results indicate that microplate-based carbohydrate arrays can be efficiently fabricated by covalent immobilization of intact carbohydrates on sol-gel-coated microplates. The microplate-based carbohydrate arrays can be applied for screening of protein-carbohydrate interactions in a high-throughput manner.
Zou
Lan
L
School of Pharmaceutical Science, Sun Yat-Sen University, Guangzhou 510275, People's Republic of China.
Pang
Hei-Leung
HL
Chan
Pak-Ho
PH
Huang
Zhi-Shu
ZS
Gu
Lian-Quan
LQ
Wong
Kwok-Yin
KY
D002241:Carbohydrates / Q000737:chemistry*; D005782:Gels; D008832:Microchemistry / Q000379:methods; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D013499:Surface Properties
D002241:Carbohydrates; D005782:Gels