Characterization of potassium channel modulators with QPatch automated patch-clamp technology: system characteristics and performance
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Standard
Characterization of potassium channel modulators with QPatch automated patch-clamp technology: system characteristics and performance. / Kutchinsky, Jonatan; Friis, Søren; Asmild, Margit; Taboryski, Rafael; Pedersen, Simon; Vestergaard, Ras K; Jacobsen, Rasmus B; Krzywkowski, Karen; Schrøder, Rikke L; Ljungstrøm, Trine; Hélix, Nathalie; Sørensen, Claus B; Bech, Morten; Willumsen, Niels.
I: ASSAY and Drug Development Technologies, Bind 1, Nr. 5, 2003, s. 685-93.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
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 -
ID: 13525343