Immobilized Proteins In Drug Discovery

一种基于目标受体、跨膜受体（细胞、核或线粒体）在流动色谱系统中包含的高通量筛选新方法已经被开发出来。在这种方法中，目标蛋白被固定在固体载体上，载体被包装成一个小柱。测试化学物质通过柱，通过固定的靶标，化合物从柱的开始到柱的结束所花费的时间与靶标和化合物之间相互作用的强度直接相关，即配体-受体复合物的结合亲和力。使用这种方法，复杂的化学和生物混合物可以在与疾病相关靶标相互作用和不相互作用的化合物之间快速分类。与此同时，与靶标结合的化合物本身在低、中、高亲和力的结合剂之间迅速分类。因此，该方法可以快速提供信息量大、信息量高的数据。我们开发了含有各种亚型尼古丁受体的柱来筛选烟草烟雾凝聚物。

A new method for high throughput screening has been developed based upon the inclusion of the target receptors, transmembrane receptors (cellular, nuclear or mitochondrial) in a flowing chromatographic system. In this approach, the target protein is immobilized on a solid support and the support packed into a small column. The test chemicals are passed through the column, over the immobilized target, and the time that it takes for the compounds to pass from the beginning of the column to its end is directly related to the strength of interaction between the target and the compound, i.e. the binding affinity of the ligand-receptor complex. Using this method, complex chemical and biological mixtures can be rapidly sorted between compounds that interact and do not interact with the disease-related target. At the same time, the compounds that bind to the target are themselves rapidly sorted between low, medium and high affinity binders. Thus, the method quickly provides a large amount of data with high information content. We have developed columns containing various subtypes of the nicotinic receptor to screen tobacco smoke condensates. The general approach has also been expanded to the immobilization of SIRT6 protein onto the surface of an open tubular capillary and on the surface of magnetic beads. The SIRT6-MBs were used to extract small molecules that bind to SIRT6 from chemical and botanical mixtures. Additional SIRT6 activity assays were developed and optimized for the identification of SIRT6 activators and SIRT6 inhibitors. Using this approach, several novel SIRT6 activators have been identified from complex matrixes. And the pharmacological activity was used to create a pharmacophore model. More recently, rhBri2 was also immobilized onto the surface of magnetic beads. The Bri2-immobilized proteins was used to identify interacting proteins from a complex matrix.