Fully Automated High Throughput Ion Channel Screening
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Process Analysis & Automation were commissioned by Biofocus Ltd. To design and build an automated multi-atomic absorption spectrometer system. Using four Thermo Electron SOLAAR AA spectrometers (with associated Gilson autosamplers), a Hamilton Microlab SWAP robot and a Kendro stacker, a fully automated and reliable workcell was produced. OVERLORD dynamic scheduler (Overlord2) was used control the workcell, with NetLORD used to control the four AA control PCs. We believe that this is the only fully commercially available automated AA system.
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Determination of protein concentration by Bradford method.pptx
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Fully Automated High Throughput Ion Channel Screening
- 1. Fully Automated High Throughput Ion Channel Screening July 2003 Adrian Kinkaid, PhD Head of Biology 1 BioFocus plc.
- 2. Drug Discovery with Vision Essential expertise for gene to pre-clinical drug discovery Assay Development and Screening Medicinal and Synthetic Chemistry Chemoinformatics and Bioinformatics
- 3. Drug Discovery with Vision collaborative, target-based drug discovery programmes Target Selection Assay Development Discovery Library Access HTS Hit-to-Lead Expansion Lead Optimisation Diverse Library Synthesis Targeted Library Synthesis Library design Molecular Modelling Data Analysis Bioinformatics flexible options for customised projects
- 4. BioFocus’ 3 UK Science Centres Cambridge Science Park Assay Development HTS Bioinformatics Chesterford Park Synthetic Chemistry Computational Chemistry Business Development UK Offices Sittingbourne Research Centre Synthetic Chemistry Computational Chemistry Headquarters and Registered Offices Total staff = 180
- 9. Summary of Ion Channel Platforms Redistribution of High Medium Low Compound voltage-dependent dyes Interference FRET-based technology High Medium/High High Radioligand binding High Low Low Non-functional/ Radioactive Automated two-electrode Low/Medium High High Low efficacy voltage clamp Automated whole-cell Low/Medium High High Cell dialysis patch clamp Planar patch clamp Medium/High High High Cell dialysis Radiometric ion flux High Medium Low Radioactive Non-radiometric ion flux High Medium Low Throughput Information quality Cost Comments
- 11. K + ATPase HERG Rb + Loading Inhibitor K + ATPase HERG K + ATPase HERG Pre-Incubation Inhibitor K + ATPase HERG K + ATPase HERG Stimulus DEPOLARISATION Rb + Flux Assay Theory Radiometric: Cerenkov counting of 86 Rb + flux Non-radiometric: atomic absorption spec. of 85 Rb + flux
- 13. Sample Processing Hollow cathode lamp source Spray chamber and nebulizer Flame Monochromator Processing electronics Data processing and instrument control Photomultiplier detector
- 15. Theory of Atomic Spectroscopy Energy n=1 n=2 n=3 n=4 Ground state Light Beer’s Law: Absorbance Atom Concentration Excitation
- 16. Theory of Atomic Emission Spectroscopy Energy n=1 n=2 n=3 n=4 Ground state Light Beer’s Law: Emission Atom Concentration Emission
- 21. IC 50 =90 nM IC 50 =102 nM Radiometric and non-radiometric flux assays are equivalent Comparison of radiometric and non-radiometric flux % Inhibition
- 22. hERG blocker dose-response curves E4031, Cisapride, Terfenadine, Risperidone, Astemizole, Haloperidol E4031 Risperidone Terfenadine Astemizole Haloperidol Cisapride
- 25. High Throughput Ion Channel Screening Platform: Reader platform initial design SOLAAR S AAS #1 SOLAAR S AAS #2 SOLAAR S AAS #3 SOLAAR S AAS #4 AutoSampler 2 Position #1 AutoSampler 2 Position #2 AutoSampler 2 Position #3 AutoSampler 2 Position #4 Linear Track Robotic arm 80 DWB On-line Storage Operating system e.g. Overlord Data Processing Activity Base All equipment must be “off the shelf”
- 26. High Throughput Ion Channel Screening Platform: Reader platform
- 27. High Throughput Ion Channel Screening Platform: Reader platform
- 28. High Throughput Ion Channel Screening Platform: Reader platform
- 30. AAS Results Correlate With Electrophysiology K + Channel
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