基于FP法的EPOR配体高通量筛选模型

A high-throughput fluorescent polarization model for screening EPO receptor ligand

目的 寻找疗效好、副作用小的EPO功能类似物。方法 用FITC标记EPO，从FVA细胞中提取含EPO受体的细胞膜，建立了用FP法筛选EPOR配体的模型，并对实验条件进行了优化和可能影响模型的因素（DMSO）进行了分析，此外用Z’因子对此模型进行了评估。结果 样品中含0％～5％DMSO不会对实验结果产生影响，Z’因子的值为0．78．结论 建立的筛选模型稳定性好和准确率高。

Aim To discover lead compound whose function is similar to EPO. Methods A high-throughput Fluorescent Polarization（FP） assay was established for screening ligand of EPO receptor from small molescules and natural products. In the FP assay, EPO receptor was abstracted from FVA ceil. EPO was labeled with FITC. Results The optimized concentration of FITC-EPO was 10^-9 mol · L^-1. Different concentrations of DMSO （VDMSO/Vtotal： 0%, 0.5%, 1%, 2.5%, 5%） didn＇t affect the FP assay. The value of Z＇ in the FP assay was 0. 78. Conclusion The assay was robust and of high sensitivity.