硫磺素T微孔板检测法筛选β-折叠蛋白聚集体亲和配体  

Screening of β-sheet protein aggregate affinity ligand by thioflavin T microplate assay

作　　者：樊雪 王晓雪 程妍[1] FAN Xue;WANG Xiaoxue;CHENG Yan(West China School of Pharmacy,Sichuan University,Chengdu,Sichuan,610041 P.R.China)

机构地区：[1]四川大学华西药学院,四川成都610041

出　　处：《华西药学杂志》2023年第3期255-257,共3页West China Journal of Pharmaceutical Sciences

摘　　要：目的采用硫磺素T微孔板检测法筛选亲和β-折叠蛋白聚集体的配体。方法将硫磺素T、β-折叠蛋白聚集体、配体混合后,置室温下孵育。在酶标仪96孔板内测定荧光强度,激发和发射波长分别为440、480 nm。计算实验组与对照组荧光强度的比率(I),筛选亲和配体。结果完成60个配体的检测。其中,有38个配体对Tau聚集体的I值小于1,有12个配体对Aβ_(1-42)聚集体的值小于1。结论硫磺素T微孔板检测法简便快捷,为相关蛋白聚集体亲和配体的初步筛选提供了有效工具。OBJECTIVE To screen the β-sheet protein aggregates affinity ligand by thioflavin T microplate assay.METHODS The samples were incubated at room temperature after mixing thioflavin T,β-sheet protein aggregates and ligands.The fluorescence intensity was measured in a 96-well plate by microplate reader with excitation and emission wavelengths set at 440 nm and 480 nm,respectively.The ratio of fluorescence intensity(I)between experiment group and control group was calculated to screen ligand affinity.RESULTS 60 ligands were evaluated.Among them,38 ligands had I values below 1 for Tau aggregates and 12 ligands had I values below 1 for Aβ_(1-42) aggregates.CONCLUSION The facile thioflavin T microplate assay provides an effective tool to screen ligands for β-sheet protein aggregates.

关 键 词：药物筛选 β-折叠蛋白聚集体 硫磺素T 微孔板检测 配体 相互作用 荧光 比率 

分 类 号：R915[医药卫生—微生物与生化药学]