Cardiac Ion Channel Panel™ Assays using Manual Patch Clamp (GLP and non-GLP)
Profiles the most important cardiac ion channels and compares concentration-response parameters
Conventional manual patch clamp assays provide the most versatile and accurate assessment of drug interactions with cardiac ion channels. For instance, continuous superfusion of cells in manual patch clamp improves the accuracy of testing difficult (e.g., “sticky”) compounds versus automated patch clamp. Moreover, recordings can be performed at physiologic temperature and in compliance with GLP guidelines. The IC50 concentration-response assays (either GLP or non-GLP) provide important benchmarks for structure-activity evaluation and cardiac risk assessment, whereas the non-GLP Screen (block comparator assay) tests compounds within structural or therapeutic groups against one or more ion channels.
Assay Features:
- Gold-standard, functional cell-based ion channel assay
- Quantitative profiling
- Provides mechanisms
- One week (non-GLP) or three weeks (GLP) turnaround times
- Physiologic temperature available for most assays
- Range-finding pre-test available for concentration-response assays
- HPLC concentration-verification available
| Channel | GLP |
non-GLP |
Positive Contol |
|---|---|---|---|
| Cav1.2 (L-type) | X |
X |
Nifedipine |
| Cav3.2 (T-type ) | X |
Ni2+ | |
| HCN2 | X |
Zatebradine | |
| HCN4 | X |
Zatebradine | |
| hERG | X |
X |
Cisapride or Terfenadine |
| Kir2.1 | X |
Ba2+ | |
| Kir3.1/3.4 | X |
Ba2+ | |
| Kir6.2/SUR2A | X |
Glybenclamide | |
| Kv1.5 | X |
X |
4-Aminopyridine |
| Kv4.3 | X |
X |
Flecainide |
| KvLQT1/minK | X |
X |
Chromanol-293B |
| Nav1.5 | X |
X |
Lidocaine |
| Assay Parameters: | IC50 (GLP) | IC50 (non-GLP) |
Screen (non-GLP) |
|---|---|---|---|
| Platform | Manual patch clamp |
Manual patch clamp |
Manual patch clamp |
| Number of concentrations | 4 |
4 |
1 - 2 |
| Number of replicates per concentration | 3 |
3 |
2 - 3 |
| Positive control (# of measurements) | 2 |
2 |
2 |
| Vehicle control (1 concentration N=3) | + |
optional |
optional |
| Assay temp: room (RT) or physiological (PT) | RT or PT |
RT or PT |
RT |
| Concentration range-finding pre-test | + |
+ |
- |
| Protocol & report | GLP |
GLP-type |
+ |
| Cumulative curves | + |
+ |
- |
| Collection of analytical samples | + |
optional |
- |
| Electronic (pdf) report provided | + |
optional |
+ |
HERG concentration-response assay
A: Superimposed patch clamp recordings (upper section) obtained before (control) and during application of 10 and 100 nM cisapride. At the end of the recording period 500 nM E-4031, a selective hERG channel blocker, was applied. The lower section of the figure shows the stimulus voltage pattern.
hERG concentration-response assay.
B: Peak current amplitude versus time during repetitive stimulation at 0.1 pulses/second.
hERG concentration-response assay.
C: Concentration-response relationship. Mean ± SEM, n = 4 cells/concentration. IC50 = 0.0234 ?M.
Optional assays:
- Dose Solution Analysis – GLP & non-GLP; verify applied concentrations with analytical sample testing of doses
- Use-dependence analysis
- Temperature-dependence analysis
Additional options:
- Additional copies of final report
- Copies of solution worksheets & data-collection forms
- Integrated Risk Assessment Summary – dose-response, repolarization, in vivo data compiled in one report
For a quote or additional information, please email inquiries@chantest.com or call 216-332-1665.
Additional Cardiac Safety Assays:
