VX-680 KINOMEscan

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（KINOMEscan assay platform）基于竞争性结合检测原理，针对单种目标化合物，与一组共计442种激酶分别进行结合检测。该检测体系包含三个核心组分：携带激酶标签的噬菌体（kinase-tagged phage）、受试化合物，以及可与激酶竞争结合位点的固定化配体（immobilized ligand）。通过对噬菌体携带的DNA标签进行定量PCR（quantitative PCR），可定量检测与固定化配体结合的激酶总量。针对各化合物报告的Kd值（单位：纳摩尔nM），通过对受试化合物进行11次三倍系列稀释，并设置二甲基亚砜（DMSO）对照组计算得到。阴性结果指受试化合物存在时未对激酶与配体的结合产生抑制作用，而较低的Kd值则代表化合物具有较强的抑制活性。