Development and evaluation of high throughput functional assay methods for HERG potassium channel.
Three functional hERG channel assay methods have been developed and evaluated. The methods were tested against five known hERG channel inhibitors: dofetilide, terfenadine (Seldane), sertindole (Serdolect), astemizole (Hismanal), and cisapride (Propulsid). The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds. The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits. The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate.
Tang
W
W
Department of Profiling and High Throughput Screening, Aventis Pharmaceuticals, Bridgewater, NJ 08807-2854, USA. weimin.tang@aventis.com
Kang
J
J
Wu
X
X
Rampe
D
D
Wang
L
L
Shen
H
H
Li
Z
Z
Dunnington
D
D
Garyantes
T
T
D000818:Animals; D016466:CHO Cells; D027682:Cation Transport Proteins; D003116:Color; D006224:Cricetinae; D051638:Ether-A-Go-Go Potassium Channels; D008564:Membrane Potentials; D008722:Methods; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D012413:Rubidium / Q000378:metabolism; D012680:Sensitivity and Specificity
D027682:Cation Transport Proteins; D051638:Ether-A-Go-Go Potassium Channels; C092509:KCNH6 protein, human; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D012413:Rubidium