A high-throughput HERG potassium channel function assay: an old assay with a new look.
In this paper, we describe an assay using radioactive rubidium (86Rb) efflux to screen functional human ether-a go-go-related gene (HERG) K+ channels in a high-throughput screening (HTS) format. This assay offers an alternative way to examine junctional interactions between chemical compounds and HERG K+ channels. Follow-up experiments and discussions were carried out to address a variety of factors that affect potency evaluation within the Rb efflux assay. Factors that can affect the assay results, such as assay time, efflux rate, and compound blocking kinetics, are discussed in detail. Our results provide some explanations for the variances of the assay results and offer some guidelines for using the Rb efflux assay to evaluate compound interactions with HERG K+ channels in the pharmaceutical industry.
Cheng
Charles S
CS
Department of Molecular Biology, Neurogen Corporation, Branford, Connecticut 06405, USA.
Alderman
Dawn
D
Kwash
Jennifer
J
Dessaint
John
J
Patel
Rita
R
Lescoe
Mary Kay
MK
Kinrade
Michele Bennett
MB
Yu
Weifeng
W
D001681:Biological Assay / Q000379:methods; D027682:Cation Transport Proteins; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004353:Drug Evaluation, Preclinical; D004345:Drug Industry; D051638:Ether-A-Go-Go Potassium Channels; D017408:Guidelines as Topic; D007700:Kinetics; D015221:Potassium Channels / Q000187:drug effects* / Q000494:pharmacology; D024642:Potassium Channels, Voltage-Gated; D012414:Rubidium Radioisotopes
D027682:Cation Transport Proteins; D018076:DNA, Complementary; D051638:Ether-A-Go-Go Potassium Channels; C092509:KCNH6 protein, human; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D012414:Rubidium Radioisotopes