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In this paper,spectroscopy fluorescence correlation spectroscopy(FCS).The principle of this method(shown in Figure 1)is mainly based on the competition of candidate drugs to the fluorescent probe-target complexes and the excellent capacity of FCS for sensitively distinguishing the free fluorescent probes and the fluorescent probe-target complexes in solution.In this study,the screening of protein kinase inhibitors was used as a model,tyrosine-protein kinase ABL1was used as a target and a known inhibitor dasatinib derivative labeled with fluorescent dyewas used as a fluorescent affinity probe.We firstly established the theoretical model of drug screening based on the binding process of fluorescent probes and targets,the competition of candidate drugs to the fluorescent probe-target complexes and FCS theory.And then,the dasatinib derivatives were synthesized and labeled with fluorescent dye Alexa488,the binding and dissociation processes of Alexa488-dasatiniband ABL1 were syste matically investigated.The dissociation constant and dissociation rate forAlexa488-dasatinib-ABL1 complex were determined.Finally,the established method was used to screen candidate drugs.The dissociation constants of ABL1 kinase to six known drugs for treating chronic myeloid leukemia(CML)were evaluated and the results obtained are well consistent with the reported values.Further more,a homemade chip with micro-wells was successfully utilized in FCS measurements as the carrier of samples,and the sample requirements were only 1~2 μ Lin this case.Our results demonstrated that the drug screening method described here is universal,sensitive and small sample and reagent requirement.We believe that this method will become a high throughput platform for screening of small molecule drugs.