OCT 23, 2018 7:00 AM PDT

FLIPR 1536-well application in high throughput screening

Sponsored by: Molecular Devices
C.E. Credits: P.A.C.E. CE Florida CE
Speakers
  • Chief Operations Officer, PPSC
    Biography
      Helma van den Hurk is currently employed as Chief Operations Officer at PPSC. She has a solid background in Medical Biology in fields ranging from neurology, developmental biology and diabetes, to innate immunity and cancer. While working towards her doctorate (1996-2001) on Alzheimer's disease and a subsequent post-doctorate on renal cell carcinomas (2000-2006), she gained ample experience in cell biology, molecular biology and biochemistry. From 2006 to 2013, she worked as Head of Research & Development at Hycult Biotech, a Dutch Biotech company focused on assays, antibodies and proteins in the field of innate immunity. In 2013 she joined PPSC and contributed to the success of the European Lead Factory as WP-lead Assay Development. Since the start of PPSC end 2012 over 200 assays have been developed and implemented contributing to over 100 screening campaigns, of which >90 % was performed in 1536-well format.
    • Assay Development Scientist, PPSC
      Biography
        Tsang Wai Lam is currently employed as Assay Development Scientist at PPSC. After receiving his BSc. in biochemistry (2001) he started working in different research departments at the Dutch pharmaceutical company Organon. There he was a member of different H2L teams where he was responsible for developing, optimizing and automating different types of biochemical/cellular assays and managing a small group of technicians for routine assays. For a brief period he worked at the Quality Control department where he was responsible for developing and optimizing cellular assays for product release. Early 2013 he joined a strong team of scientist at PPSC were he was responsible for the assay development/optimization and screening of different projects in the European Lead Factory.

      Abstract

      DATE: October 23, 2018
      TIME: 7:00AM PDT
      10:00AM EDT, 3:00PM BST, 4:00PM CET

      Over the past five years, over 70 high-throughput screening campaigns have been performed at Pivot Park Screening Centre (PPSC) for the European Lead Factory (ELF).  These HTS campaigns have been executed on a compound library that has grown from 320,000 to 450,000 compounds.

      The screens cover a wide range of target classes, including more demanding cellular targets like ion channels and protein-protein interactions. As an example of one of the ELF screening campaigns, we will present the results of a high-throughput screening to identify selective agonists for an ion channel using the Molecular Devices FLIPR Tetra® High-Throughput Cellular Screening System.

      In this project, the assay was based on stable cell lines expressing the ion channel. The assay was successfully miniaturized to 1536 well format using a standard Molecular Devices membrane potential dye. The screening cascade will be presented.

      Another highly important aspect for effective drug development is the assessment of cardiac safety and efficacy of drug candidates. Hence, there is a pressing need for in vitro screening methods to detect cardioactive effects of compounds early in the drug discovery process. To provide a relevant in vitro model to investigate effects of drug candidates on cardiac physiology, we have evaluated hiPSC-derived ventricular cardiomyocytes (Pluricyte® Cardiomyocytes), which exhibit a relatively high level of maturity. We used these cardiomyocytes in combination with a fluorescent calcium dye (FLIPR Calcium 6 Assay Kit, Molecular Devices) to develop a high-throughput drug screening assay on the FLIPR Tetra® (Molecular Devices) screening platform.

      After optimization of the assay for application with 384-well plates, we investigated the impact of different cardioactive compounds (e.g. hERG-channel blockers, calcium channel agonists/antagonists and β-adrenergic agonists) on the calcium transients in Pluricyte® Cardiomyocytes. 

      Learning objectives:

      • Using full automation to perform complex GPCR testing
      • Tips and tricks for 1536-well FLIPR screening
      • Calcium kinetics in cardiac safety testing

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