TY - JOUR

T1 - Characterization of potassium channel modulators with QPatch automated patch-clamp technology: system characteristics and performance

AU - Kutchinsky, Jonatan

AU - Friis, Søren

AU - Asmild, Margit

AU - Taboryski, Rafael

AU - Pedersen, Simon

AU - Vestergaard, Ras K

AU - Jacobsen, Rasmus B

AU - Krzywkowski, Karen

AU - Schrøder, Rikke L

AU - Ljungstrøm, Trine

AU - Hélix, Nathalie

AU - Sørensen, Claus B

AU - Bech, Morten

AU - Willumsen, Niels

N1 - Keywords: Animals; Biotechnology; CHO Cells; Cell Culture Techniques; Cells, Cultured; Cricetinae; Cricetulus; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Electrophysiology; Equipment Design; Equipment Failure Analysis; Feasibility Studies; Humans; Ion Channel Gating; Kidney; Membrane Potentials; Microelectrodes; Patch-Clamp Techniques; Potassium Channel Blockers; Potassium Channels; Reproducibility of Results; Robotics; Sensitivity and Specificity

PY - 2003

Y1 - 2003

N2 - Planar silicon chips with 1-2-microm etched holes (average resistance: 2.04 +/- 0.02 MOmega in physiological buffer, n = 274) have been developed for patch-clamp recordings of whole-cell currents from cells in suspension. An automated 16-channel parallel screening system, QPatch 16, has been developed using this technology. A single-channel prototype of the QPatch system was used for validation of the patch-clamp chip technology. We present here data on the quality of patch-clamp recordings and from actual drug screening studies of human potassium channels expressed in cultured cell lines. Using Chinese hamster ovary (CHO) and human embryonic kidney cells (HEK), gigaseals of 4.1 +/- 0.4 GOmega (n = 146) and high-quality whole-cell current recordings were obtained from hERG and KCNQ4 potassium channels. Success rates for gigaseal recordings varied from 40 to 95%, and 67% of the whole-cell configurations lasted for >20 min. Cells were maintained in suspension up to 4 h in a cell storage facility that is integrated in the QPatch 16. No decline in patchability was observed during this time course. A series of screens was conducted with known inhibitors of the hERG and KCNQ4 potassium channels. Dose-response relationship characterizations of verapamil and rBeKm-1 blockage of hERG currents provided IC(50) values similar to values reported in the literature.

AB - Planar silicon chips with 1-2-microm etched holes (average resistance: 2.04 +/- 0.02 MOmega in physiological buffer, n = 274) have been developed for patch-clamp recordings of whole-cell currents from cells in suspension. An automated 16-channel parallel screening system, QPatch 16, has been developed using this technology. A single-channel prototype of the QPatch system was used for validation of the patch-clamp chip technology. We present here data on the quality of patch-clamp recordings and from actual drug screening studies of human potassium channels expressed in cultured cell lines. Using Chinese hamster ovary (CHO) and human embryonic kidney cells (HEK), gigaseals of 4.1 +/- 0.4 GOmega (n = 146) and high-quality whole-cell current recordings were obtained from hERG and KCNQ4 potassium channels. Success rates for gigaseal recordings varied from 40 to 95%, and 67% of the whole-cell configurations lasted for >20 min. Cells were maintained in suspension up to 4 h in a cell storage facility that is integrated in the QPatch 16. No decline in patchability was observed during this time course. A series of screens was conducted with known inhibitors of the hERG and KCNQ4 potassium channels. Dose-response relationship characterizations of verapamil and rBeKm-1 blockage of hERG currents provided IC(50) values similar to values reported in the literature.

U2 - 10.1089/154065803770381048

DO - 10.1089/154065803770381048

M3 - Journal article

C2 - 15090241

VL - 1

SP - 685

EP - 693

JO - Assay and Drug Development Technologies

JF - Assay and Drug Development Technologies

SN - 1540-658X

IS - 5

ER -