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Nanion Introducing:

Nanion Introducing:. SyncroPatch 384PE - the PatchEngine . Superior Ion Channel Drug Screening. Nanion:. 11 years of patch clamping - and more to come. The Port-a-Patch. The world’s smallest patch clamp rig. Fast access to high quality patch clamp data

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Nanion Introducing:

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  1. Nanion Introducing: SyncroPatch 384PE - the PatchEngine. Superior Ion Channel Drug Screening.

  2. Nanion: 11 years of patch clamping - and more to come • The Port-a-Patch. • The world’s smallest patch clamp rig. • Fast access to high quality patch clamp data • Quick evaluation of cells and compounds • Novel experimental possibilities • The SURFE2R. • Catch the wave for transporters. • In-depth analysis of transporter protein activity and function • Compatible with diverse membrane sources • Multiple targets investigated with one sensor • The Patchliner. • Because quality does matter. • Unlimited experimental freedom • Best of all worlds: throughput, performance and versatility • Press one button and walk away: - 48 cells in one run Measure More Membrane • The CardioExcyte 96. • Bump up your safety screening. • Ultra-precise impedance measurements • Real time access to beating parameters • Quick experiments and long term observations • The Orbit 16. • Instant bilayer – just add protein. • 16 parallel bilayers • Low noise, high bandwidth recordings • Compatible with your existing amplifier • The SyncroPatch 96. • Get more throughput. • Cost-efficient ion channel screening • Ligand- and voltage-gated channels • High throughput and high data quality • The Vesicle Prep Pro. • Liposomes made easy. • Quick and easy formation of GUVs • Temperature control • Stable bilayers for ephys recordings Smart tools for ion channel and transporter research and screening

  3. The PatchEngine– a Revolutionary Instrument for APC-based Ion Channel Drug Screening • Executive summary • 384 or even 768 cells in parallel (modular approach) • Up to 20,000 data points per day • Reliable data from diverse ion channels targets • Integrates easily into an HTS environment • Cost per data point: € 0.2 • Available for shipping NOW!

  4. The PatchEngine– a Revolutionary Instrument for APC-based Ion Channel Drug Screening • Key features • 384 or 768 cells in parallel (384 or 768 amplifier) • 384 pipettor head • 85 % success rates routinely obtained • Excellent results with voltage-gated & ligand-gated ion channels • Modular approach – easy upgrade & integration • Single- and multi-hole recording substrates

  5. The PatchEngine– modular, compact and accessible 96 48 Higher throughput Patch Clamping 16 384 • Stand alone machines The PatchEngine– the core of the SyncroPatch 384PE • Integration in HTS? 8

  6. The PatchEngine– 100 % integration in HTS environment: • Fits into commercially available liquid handler • Up to two PE’s per robot • Open design allows integrations into fully robotic environments • Used successfully with • Beckman Coulter’s Biomek The PatchEngine– the core of the SyncroPatch 384PE

  7. SyncroPatch 384PE–Set-up overviewusing the Biomek FX robot • Components: • BiomekFX with a 384 pipetting head including gripper • 1-2 SyncroPatch 384PEModules • 1 - 2 x 384 channel amplifier • SyncroPatch 384 software • NPC-384 borosilicate chip • Temperature controlled cell hotel • Integrated Barcode scanner • Optional 8 channel pipettinghead (Screen 100% of all compounds) Open design allows for seamless integration into robotic HTS-highways

  8. Integration of the SyncroPatch 384PE into HTS environment PlateHotel FluorescenceReader Multidrop Cytomat Incubator PlateWasher PlateHandler SealRemover PlateSealer Shuttle ThermoShaker TipLoader SP 384PE Module 1 SP 384PE Module 2

  9. Integration of the SyncroPatch 384PE into HTS environment HTS Software Scheduling

  10. Software integration of the SyncroPatch 384PE into the Biomek Software

  11. Overview Experiment Window Close up selected traces Wellplate overview Experiment statistics One well highlighted CHO-hKv1.3 cellswere kindlyprovidedbyEvotec

  12. Data Control 384–Outstanding data handling and export • DataControl 384 – intuitive& easy-to-use featuring: • Intuitive, quick & easy-to-use setup of exps • Powerful data analysis functions • Compound view • Automated data export • PDF-reporting Raw data traces of 384 hKv1.3 current responses

  13. SyncroPatch 384 PE–Assay compatibility examples • The SyncroPatch384 PE has been validated with: • CHO, HEK, LTK and RBL-1 cells • A variety of voltage-gated ion channels • includinghNav1.7, hKv1.3, Kir2.1 • A variety of ligand-gated ion channels • includingincludinghGABAAα1,β2,γ2 , hGlycineα1

  14. SyncroPatch 384 PE–Consumables NPC-384 Available as single and multi-hole borosilicate plates: Four-hole substrate: Single-hole substrate:

  15. Current Voltage Relationship of hKv1.3 expressed in CHO cells CHO-hKv1.3 cellswere kindlyprovidedbyEvotec

  16. Current Voltage Relationship of hKv1.3 expressed in CHO cells CHO-hKv1.3 cellswere kindlyprovidedbyEvotec

  17. hKv1.3 block by Quinidine CHO-hKv1.3 cellswere kindlyprovidedbyEvotec

  18. hKv1.3 block by Quinidine (5 µM, 50 µM and 500 µM ) Exp length: 16 minutes Success rate: 85.2 % Quinidine IC50: 18.9µM CHO-hKv1.3 cellswere kindlyprovidedbyEvotec

  19. Stability of the seal resistance during compound application Four additions took ca 500 seconds Success rate: 85.2 % Fourapplications: Onecontrolwashandsubsequently threeconcentrations

  20. Current Voltage Relationship of hNav1.7 expressed in CHO cells CHO-hNav1.7 cellswere kindlyprovidedbyAnaxon

  21. Current Voltage Relationship of hNav1.7 expressed in CHO cells CHO-hNav1.7 cellswere kindlyprovidedbyAnaxon

  22. hNav1.7 block by Tetracaine (5 µM, 50 µM and 500 µM ) CHO-hNav1.7 cellswere kindlyprovidedbyAnaxon

  23. hNav1.7 block by Tetracaine (5 µM, 50 µM and 500 µM) Exp length: 15 minutes Success rate: 86.2 % Tetracaine IC50: 49.6µM CHO-hNav1.7 cellswere kindlyprovidedbyAnaxon

  24. Fluidic Properties tested on RBL cells Exp length: 15 minutes Success rate: 71,1 % Exchange rate: <50 ms PotassiumcurrentsthroughendogenousKir 2.1-channels in RBL cells

  25. hGABAAα1,β2,γ2expressed in HEK cells ThreeGABA-applications (5 µM) using multi-hole chip

  26. Dose Response CurveofGlycine on the human α1-Glycine Receptor LTK cells expressing human 1-glycine receptor Glycine (100 µM, 300 µM, 1 mM) was applied using a stacked application approach A multi-hole chip was used. Cells were kindly provided by AstraZeneca.

  27. SyncroPatch 384 PE–quick priming and quick experiments Experiment overview Chip load & Priming 1-2 control washes + 3 cmpd concentrations Exp end & Chip unload 5 minutes 8 – 12 minutes 2 minutes Total experiment length for 4 – 5 additions: 15 – 20 minutes Experiment length is exposure time dependent.

  28. SyncroPatch 384 PE–Assay compatibility examples • The SyncroPatch 384 PE assay validation: • Assay development straightforward, directly adaptable from other platforms • Whole-cell parameters are stable throughout the experiment • Success rates for completed experiments are routinely ~ 85% • Compatible with voltage- and ligand-gated ion channels Wrap up data section

  29. Nanion Technologies – 10 years ago introducing the Port-a-Patch Reducing a set-up that takes up an entire room to something so small that you can hold in your hand. • The Port-a-Patch • Makes patch clamp accessible to non-experts • Increases throughput 5 – 10 x • Allows quick tests of compounds and cells

  30. Nanion Technologies – still increasing the throughput & reducing size 384 768 • Each PatchEngine: • 384 cells in parallel • 2 units possible • Integrates easily in HTS

  31. SyncroPatch384PE – Superior Ion Channel Drug Screening • Giga-seal recordings • 384 or even 768 simultaneous whole-cell recordings • Multiple units per liquid handler • High success rates - 85 % • Up to 20,000data points per day • Voltage- and ligand-gated ion channels • Competitive price per data point ~ €0.20 • Customized hole sizes and multi-hole chips … Customized to your HTS environment

  32. Nanion Customer Reference - a global customer base distributed between industrial and academic laboratories • Companies: • Abbott • Almirall Prodesfarma • AstraZeneca • Amgen • Aurigene Discovery Technologies, Ltd. • Axiogenesis • Bayer • Bionomics • Boehringer Ingelheim • Bristol-Myers Squibb • Center for New Drug Safety Evaluation and Research (NCDSER), • Chromocell • conoGenetix biosciences GmbH • Evotec • Genionics • GlaxoSmithKline • Grünenthal • Hofman La Roche • Hydra Biosciences • Japanese Tobacco • Lupin Limited • Merck • Merch Serano • Novartis • Novo Nordisk • Nycomed • Ono Pharmaceutical • Orion Pharma • Pfizer • Pierre Fabre • Regeneron Pharmaceuticals • Rytmion • Sanofi-Aventis • Shinogi • Siena Biotech • SK Holding, Co Ltd • Sundia MediTech Company, Ltd. • Suven Life Sciences Ltd, Hyderabad, India • Syngenta • UCB Pharma • Vertex Pharmaceuticals • Vimta Labs Life Sciences, Hyderabad, India • Wyeth • Xention • Research institues and universites: • Cambridge University, Cambridge, UK • Charité - Universitätsmedizin Berlin, Germany • Consiglio Nazionale delle Ricerche, Povo (Trento), Italy • Copenhagen University, Denmark • Cornell University, Geneva, NY, USA • Eberhard Karls Universität Tübingen, Germany • FMP, Berlin, Germany • Forschungszentrum Caesar, Bonn, Germany • Forschungszentrum Jülich GmbH, Jülich, Germany • Friedrich-Schiller-Universität Jena, Germany • Georg-August-Universität Göttingen, Germany • Griffith University, Nathan, Australia • GSF - Forschungszentrum für Umwelt und Gesundheit, GmbH, Neuherberg, Germany • Hasselt University, Diepenbeek ,Belgium • Hertie Institute for Clinical Brain Research, Germany • Howard Florey Institute, USA • Howard Hughes Medical Institute, Ashburn, VA, USA • Institute of Genomics and Integrative Biology, Delhi, India • IPBS-CNRS, Tolouse, France • Istituto Italiano di Tecnologia, Genova, Italy • Jacobs University, Bremen, Germany • JNCASR, Bangalore, India • Julius von Sachs Institut, Würzburg, Germany • Karolinska Institute, Stockholm, Sweden • Kiserleti Orvostudomanyi Kutatointezet, Budapest, Hungary • Korea Institute of Science and Technology (KIST), Seoul, South Korea • Laboratorio Especial de Toxinologia Aplicada CAT-CEPID, Sao Paolo, Brazil • Ludwig Maximillian Universität, München, Germany • Lund University, Lund, Sweden • Max Planck Institut, Göttingen, Germany • Medizinische Hochschule Hannover, Germany • Research institues and universites: (cont.) • Nanjing University, Nanjing, China • Nanyang Technological University, Singapore • National University of Singapore, Singapore • National Center for New Drug Safety Evaluation and Research (NCDSER), Shanghai, China • National Heart Center, Singapore • National Jewish Medical and Research Center, Denver, CO, USA • NIH/NINDS, Bethesda, MD, USA • NIST, Gaithersburg, MD, USA • New York University, NY, USA • RMIT University, Bundoora, Australia • Ruhr-Universität Bochum, Germany • Shanghai Institute of Materia Medica, Shanghai, China • Simon Fraser University, Burnaby, Canada • Southampton General Hospital, Southampton, UK • Stiftung Alfred-Wegener-Institut für Polar- und Meeresforschung, Bremerhaven, Germany • Syracuse Univerity, NY, USA • Technische Universität Ilmenau, Germany • Technische Universität Darmstadt, Germany • Technische Universiät München, Germany • Uppsala University, Uppsala, Sweden • UNESCO, Bahía Blanca, Argentina • Universidade Federal de Sao Paulo, Brazil • Università degli Studi di Genova, Italy • Universitatea Lucian Blaga Din Sibiu, Romania • University of California, Los Angeles, CA, USA • University of California, San Diego, CA, USA • University of Cambridge, UK • University of Cincinnati, OH, USA • University of Groningen, the Netherlands • University of Leeds, UK • Universidade de Lisboa, Portugal • Research institues and universites: (cont.) • University of Michigan, Ann Arbor, MI, USA • Università degli Studi di Napoli - Federico II, Napoli, Italy • University of North Carolina, Wilmington, NC, USA • University of Queensland, Brisbane, Australia • University of Rochester, NY, USA • University of Tennessee Health Science Center, USA • University of Valladolid and Spanish National Research Council (CSIC), Valladolid, Spain • University of Washington, USA • University of Wisconsin, Madison, WI, USA • Universität Zürich, Switzerland • Universitatea Lucian Blaga din Sibiu, Romania • Universität Augsburg, Germany • Universität Freiburg, Germany • Universität für Bodenkultur, Wien, Austria • Universität Heidelberg, Germany • Universität Hohenheim, Germany • Universität Osnabrück, Germany • Vanderbilt Medical School, Nashwille, TN, USA • Virginia State University, Blacksburg, VA, USA • X-Lab, Göttingen, Germany • Yale University, New Haven, CT, USA • Inje University, Busan, South Korea • Companies: • Abbott • Almirall Prodesfarma • AstraZeneca • Amgen • Aurigene Discovery Technologies, Ltd. • Axiogenesis • Bayer • Bionomics • Boehringer Ingelheim • Bristol-Myers Squibb • Center for New Drug Safety Evaluation and Research (NCDSER), • Chromocell • conoGenetix biosciences GmbH • Evotec • Genionics • GlaxoSmithKline • Grünenthal • Hofman La Roche • Hydra Biosciences • Japanese Tobacco • Lupin Limited • Merck • Merch Serano • Novartis • Novo Nordisk • Nycomed • Ono Pharmaceutical • Orion Pharma • Pfizer • Pierre Fabre • Regeneron Pharmaceuticals • Rytmion • Sanofi-Aventis • Shinogi • Siena Biotech • SK Holding, Co Ltd • Sundia MediTech Company, Ltd. • Suven Life Sciences Ltd, Hyderabad, India • Syngenta • UCB Pharma • Vertex Pharmaceuticals • Vimta Labs Life Sciences, Hyderabad, India • Wyeth • Xention

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