Microarray-based study of carbohydrate-protein binding by gold nanoparticle probes.
In order to develop a novel high-throughput tool for monitoring carbohydrate-protein interactions, we prepared carbohydrate or glycoprotein microarrays by immobilizing amino modified carbohydrates on aldehyde-derivatized glass slides or glycoprotein on epoxide-derivatized glass slides and carried out lectin binding experiments by using these microarrays, respectively. The interaction events are marked by attachment of gold nanoparticles followed by silver deposition for signal enhancement. The attachment of the gold nanoparticles is achieved by standard avidin-biotin chemistry. The detection principle is resonance light scattering (RLS). The well-defined recognition systems, namely, three monosaccharides (Man-alpha, Glc-beta and Gal-beta) or three glycoproteins (Asf, RNase A and RNase B) with two lectins (ConA and RCA120), were chosen here to establish the RLS assay, respectively. Highly selective recognition of carbohydrate-protein down to 25.6 pg/mL for RCA120 in solution and 8 microM for Gal-beta and 32 ng/mL for Asf on the microarray spots is demonstrated.
Gao
Jingqing
J
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China.
Liu
Dianjun
D
Wang
Zhenxin
Z
D000818:Animals; D032449:Canavalia / Q000378:metabolism; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism; D006046:Gold; D037102:Lectins / Q000032:analysis / Q000378:metabolism; D008027:Light; D053768:Metal Nanoparticles; D046228:Microarray Analysis / Q000379:methods*; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism; D011485:Protein Binding; D012680:Sensitivity and Specificity
D002241:Carbohydrates; D006023:Glycoproteins; D037102:Lectins; D010940:Plant Proteins; D006046:Gold