Landry James P, Fei Yiyan, Zhu X D
Department of Physics, University of California at Davis, Davis, California, 95616, USA.
Int Drug Discov. 2011 Dec:8-13.
Small-molecule compounds remain the major source of therapeutic and preventative drugs. Developing new drugs against a protein target often requires screening large collections of compounds with diverse structures for ligands or ligand fragments that exhibit sufficiently affinity and desirable inhibition effect on the target before further optimization and development. Since the number of small molecule compounds is large, high-throughput screening (HTS) methods are needed. Small-molecule microarrays (SMM) on a solid support in combination with a suitable binding assay form a viable HTS platform. We demonstrate that by combining an oblique-incidence reflectivity difference optical scanner with SMM we can screen 10,000 small-molecule compounds on a single glass slide for protein ligands without fluorescence labeling. Furthermore using such a label-free assay platform we can simultaneously acquire binding curves of a solution-phase protein to over 10,000 immobilized compounds, thus enabling full characterization of protein-ligand interactions over a wide range of affinity constants.
小分子化合物仍然是治疗性和预防性药物的主要来源。针对蛋白质靶点开发新药通常需要筛选大量具有不同结构的化合物，以寻找对靶点具有足够亲和力和理想抑制效果的配体或配体片段，然后再进行进一步的优化和开发。由于小分子化合物数量众多，因此需要高通量筛选（HTS）方法。固体支持物上的小分子微阵列（SMM）与合适的结合测定相结合，形成了一个可行的HTS平台。我们证明，通过将斜入射反射率差光学扫描仪与SMM相结合，我们可以在单个载玻片上筛选10,000种小分子化合物以寻找蛋白质配体，而无需进行荧光标记。此外，使用这种无标记测定平台，我们可以同时获得溶液相蛋白质与超过10,000种固定化化合物的结合曲线，从而能够在广泛的亲和力常数范围内全面表征蛋白质-配体相互作用。
