State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, P. R. China.
Analyst. 2011 Oct 21;136(20):4301-7. doi: 10.1039/c1an15230k. Epub 2011 Sep 1.
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) NH(2)-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-α, Glc-α and Gal-β) 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,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 (IC(50))) for inhibitors in solution are also demonstrated by the saccharide competing assays.
这里，建立了一种基于三维（3D）碳水化合物微阵列的等离子体共振光散射（RLS）分析方法，用于研究碳水化合物-凝集素结合的高选择性。3D 碳水化合物微阵列是通过将氨基修饰的碳水化合物固定在自制的第四代（G4）NH（2）末端聚（酰胺-胺）树枝状大分子（PAMAM）修饰的基底上而制备的。通过生物素-亲和素反应，用 13nm 肽稳定的金纳米粒子标记碳水化合物-凝集素结合事件后，3D 微阵列可以通过 RLS 扫描仪直接检测，而无需传统的银增强步骤。这里选择了三个单糖（Man-α、Glc-α 和 Gal-β）和两个凝集素（Con A 和 RCA 120）的明确识别系统，分别建立了 RLS 分析方法。实现了表面碳水化合物和凝集素的表面离解常数（K（D，surf））的定量测定。此外，通过糖竞争测定还证明了溶液中抑制剂的抑制值（即抑制常数（K（i））和产生 50%抑制所需的抑制剂浓度（IC（50））。
