R406 KINOMEscan-2

The KINOMEscan assay platform is based on a competition binding assay that is run for a compound of interest against each of a panel of 442 kinases. The assay has three components: a kinase-tagged phage, a test compound, and an immobilized ligand that the compound competes with to displace the kinase. The amount of kinase bound to the immobilized ligand is determined using quantitative PCR of the DNA tag. Kd values (nM) reported for each compound were determined using 11 serial threefold dilutions of test compound and a DMSO control. A null result means no inhibition of kinase binding to the ligand in the presence of the compound and low Kd means strong inhibition.

KINOMEscan检测平台基于竞争性结合检测技术，可针对目标化合物与由442种激酶组成的检测阵列中的每一种激酶分别开展结合实验。该检测体系包含三个核心组分：激酶标记噬菌体（kinase-tagged phage）、受试化合物，以及可与受试化合物竞争结合并置换结合于其上的激酶的固定化配体。通过对噬菌体携带的DNA标记进行定量聚合酶链反应（quantitative PCR），可定量测定与固定化配体结合的激酶含量。针对每种化合物所报告的解离常数（Kd，单位为纳摩尔nM），通过将受试化合物进行11次三倍梯度稀释，并设置二甲基亚砜（DMSO）对照组测算得到。若检测结果为阴性，则代表在该化合物存在的条件下，激酶与配体的结合未受到抑制；而较低的Kd值则意味着该化合物对激酶结合配体具有较强的抑制作用。