Glycan array on aluminum oxide-coated glass slides through phosphonate chemistry.
A new type of glycan array covalently or noncovalently attached to aluminum oxide-coated glass (ACG) slides has been developed for studies of enzymatic reactions and protein binding. To prepare the noncovalent array, glycans with a polyfluorinated hydrocarbon (-C(8)F(17)) tail are spotted robotically onto the ACG slide surface containing a layer of polyfluorinated hydrocarbon terminated with phosphonate. After incubation and washing, the noncovalent array can be characterized by MS-TOF via ionization/desorption at a low laser energy without addition of matrix. A representative cellotetraose array was developed to study the activity and specificity of different cellulases and to differentiate the exo- and endoglucanase activities. To prepare the covalent array, glycans with a phosphonic acid tail were synthesized and spotted robotically onto the ACG slide surface. After incubation, the slides can be used directly for quantitative protein binding analysis. Compared to the preparation of glycan arrays on glass slides and other surfaces, this method of arraying using phosphonic acid reacting with ACG is more direct, convenient, and effective and represents a new platform for the high-throughput analysis of protein-glycan interactions.
Chang
Shih-Huang
SH
Genomics Research Center, Academia Sinica, 128 Academia Road, Section 2, Nankang, Taipei 115, Taiwan.
Han
Jeng-Liang
JL
Tseng
Susan Y
SY
Lee
Hsin-Yu
HY
Lin
Chin-Wei
CW
Lin
Yu-Chen
YC
Jeng
Wen-Yih
WY
Wang
Andrew H-J
AH
Wu
Chung-Yi
CY
Wong
Chi-Huey
CH
D000537:Aluminum Oxide / Q000737:chemistry*; D002480:Cellulase / Q000737:chemistry; D005898:Glass; D013058:Mass Spectrometry; D063065:Organophosphonates / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*
D063065:Organophosphonates; D011134:Polysaccharides; D002480:Cellulase; D000537:Aluminum Oxide