Journal
ANALYST
Volume 136, Issue 20, Pages 4301-4307Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c1an15230k
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Funding
- NSFC [20875087]
- Jilin Provincial Science and Technology Department [20100701]
- Chinese Academy of Sciences
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Here, a three-dimensional (3D) carbohydrate microarray-based plasmon resonance light scattering (RLS) assay has been established for studying carbohydrate-lectin binding with high selectivity. The 3D carbohydrate microarray is fabricated by immobilizing amino-modified carbohydrates on the home-made fourth-generation (G4) NH2-terminated poly(amidoamine) dendrimers (PAMAM)-modified substrate. After marking the carbohydrate-lectin binding events by 13 nm peptide-stabilized gold nanoparticles through the biotin-avidin reaction, the 3D microarray can be directly detected by the RLS scanner without the conventional silver enhancement step. The well defined recognition systems: three monosaccharides (Man-alpha, Glc-alpha and Gal-beta) with two lectins (Con A and RCA 120), have been chosen here to establish the RLS assay, respectively. Quantitative determination of the surface dissociation constants (K-D,K-surf) for surface carbohydrates and lectins has been achieved. In addition, inhibition values (i.e. the inhibition constants (K-i) and the concentrations of inhibitors required to produce 50% inhibition (IC50)) for inhibitors in solution are also demonstrated by the saccharide competing assays.
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