SpringerProtocols: Abstract: Microarray-Based Study of Carbohydrate?Protein Binding

10. Microarray-Based Study of Carbohydrate–Protein Binding

By: Zhenxin Wang¹, Jingqing Gao¹

Abstract

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To develop a novel high-throughput tool for monitoring carbohydrate–protein interactions, carbohydrate or glycoprotein microarrays have been prepared for binding with lectins. The interaction events are marked by attachment of fluorescent dyes and gold nanoparticles. The attachment of the fluorescent dyes and gold nanoparticles is achieved by standard avidin–biotin chemistry. The detection principle is fluorescence or resonance light scattering (RLS). The electroless deposition of silver onto the gold particles has been employed for RLS signal enhancement. Well-defined recognition systems, three monosaccharides (Man-α, Glc-β, and Gal-β) or three glycoproteins (Asf, RNase A, and RNase B) with two lectins (ConA and RCA120), are chosen here to establish the microarray-based assay, respectively. Highly selective recognition of carbohydrate–protein down to 25.6 pg/mL for RCA120 in solution and 8 μM for Gal-β and 32 ng/mL for Asf on the microarray spots is demonstrated.

Affiliation(s): (1) State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, and Graduate School of the Chinese Academy of Sciences, Beijing, China

Book Title: Functional Glycomics: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 600 | Pub. Date: Oct-30-2009 | Page Range: 145-153 | DOI: 10.1007/978-1-60761-454-8_10

Subject: Biochemistry

Key Words: Glycan microarray - fluorescence - resonance light scattering - gold nanoparticle

References

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