Liang Pi-Hui, Wang Sheng-Kai, Wong Chi-Huey
Department of Chemistry, the Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
J Am Chem Soc. 2007 Sep 12;129(36):11177-84. doi: 10.1021/ja072931h. Epub 2007 Aug 18.
Carbohydrate-protein interactions on surface and in solution were quantitatively measured by a glycan microarray. Assessing carbohydrate affinities is typically difficult due to weak affinities and limited sources of structurally complex glycans. We described here a sensitive, high-throughput, and convenient glycan microarray technology for the simultaneous determination of a wide variety of parameters in a single experiment using small amounts of materials. Assay systems based on this technology were developed to analyze multivalent interactions and determine the surface dissociation constant (KD,surf) for surface-coated mannose derivatives with mannose binding lectins and antibodies. Competition experiments that employed monovalent ligands in solution yielded KD and Ki values in solution similar to equilibrium binding constants obtained in titration microcalorimetry and surface plasmon resonance experiments.
通过聚糖微阵列对表面和溶液中的碳水化合物 - 蛋白质相互作用进行了定量测量。由于亲和力较弱且结构复杂的聚糖来源有限，评估碳水化合物亲和力通常很困难。我们在此描述了一种灵敏、高通量且便捷的聚糖微阵列技术，可在单次实验中使用少量材料同时测定多种参数。基于该技术开发了分析多价相互作用的检测系统，并确定了表面包被的甘露糖衍生物与甘露糖结合凝集素和抗体的表面解离常数（KD,surf）。在溶液中使用单价配体的竞争实验得到的溶液中的KD和Ki值，类似于滴定微量热法和表面等离子体共振实验中获得的平衡结合常数。
