Aim: To develop and optimize a competitive fluorescence polarization(FP)method,and use it as a high-throughput screening(HTS)assay for drug discovery.Methods: Human lectin-like oxidized low-density lipoprotein receptor-1(hLOX-1)and oxidized low-density lipoprotein(oxLDL)were used to establish a high-throughput fluorescence polarization assay to screen ligands of human LOX-1. A96-well plate assay was performed with a fast plate reader. Three fluoresceinisothiocyanate-labeled hLOX-1 concentrations(100, 200, and 400 nmol/L)wereselected to be titrated by oxLDL(from 0.05 nmol/L to 100 μmol/L)in order to obtainoptimal reactive concentrations. The concentration of Me_2SO used(0%, 1%, 3%,5%) and incubation time(15 min, 30 min, 1 h, 2 h)were optimized. The Z’factor wascalculated to estimate the quality of FP-based HTS. Results: Concentrations of200 nmol/L for human LOX-1 and 50 μmol/L for oxLDL were used in the actualassay. Concentrations of 0% to 5% Me_2SO and different reaction times did notaffect the FP-based HTS. The Z’value was 0.66. By using this detection andscreening system, 12 700 compounds were screened and 3 ligands with an IC_(50) ofless than 4.5 μmol/L were found. Conclusion: The established competitive FP-based assay is sensitive, stable, highly reproducible and robust, and suitable forHTS for ligands of the hLOX-1 receptor. Aim: To develop and optimize a competitive fluorescence polarization (FP) method, and use it as a high-throughput screening (HTS) assay for drug discovery. Methods: Human lectin-like oxidized low-density lipoprotein receptor- ) and oxidized low-density lipoprotein (oxLDL) were used to establish high-throughput fluorescence polarization assay to screen ligands of human LOX-1. A96-well plate assay was performed with a fast plate reader. Three fluorescein isothiocyanate- labeled hLOX-1 Concentrations (100, 200, and 400 nmol / L) were selected to be titrated by oxLDL (from 0.05 nmol / L to 100 μmol / L) The Z’factor wascalculated to estimate the quality of FP-based HTS. Results: Concentrations of 200 nmol / L for human LOX- 1 and 50 μmol / L for oxLDL were used in the actualassay. Concentrations of 0% to 5% Me_2SO and different reaction times did no The use of this FP-based HTS. The Z’value was 0.66. By using this detection and screening system, 12 700 compounds were screened and 3 ligands with an IC 50 (of 50) of less than 4.5 μmol / L were found. Conclusion: The established competitive FP -based assay is sensitive, stable, highly reproducible and robust, and suitable for HTS for ligands of the hLOX-1 receptor.