Transnational Call

SMALL MOLECULE SCREENING

 

2D imaging of cellular systems (spheroids, organoids from iPS or primary cells) to identify compounds interfering with cancer development or other signaling pathways.

Abstract

The Screening Unit offers image-based screening with automated microscopes in 384-well format and automated identification and quantification of up to 6 differently fluorescently labeled cellular structures. FVB-FMP can use primary cells from mice or patients and have also experience in usage of iPS cells.

Description of technology offered under this call

The Screening Unit of the FVB-FMP offers a broad portfolio of screening technologies to identify drugs or proteins (RNAi-interference, CRISPR/Cas9) involved in development of healthy or disease determined biological systems. More then 300 projects from Europe and abroad have been supported in the past. This knowledge is passed to the supported projects without any loss of IP to your project and generated results. We work with cells from patients, animal derived primary cells and organisms like C. elegans or Zebrafish embryos. For support in 384-well or other format microtiter plates we need an assay already established in your lab and help to downscale it towards lowest volumes to reduce your costs for reagents towards a minimum. We already use three ArrayScan microscopes (one embedded in a FreedomEvo Workstation and providing a climate chamber for live cell assays) and one FLIPR-Tetra system for kinetic imaging. We already organized security settings to be able to analyze human cells from hospitals. Our detection systems support all kinds of read outs with the only exception being the usage of radioactively-labeled probes. We support image-based screens providing fluorescent or luminescent readouts, but also bright-field imaging and readouts based on changes in morphology of cells or cell-groups in colonies.

Number of compounds screened

100,000 (full commercial set) of the EU-OPENSCREEN ERIC compound library

Time frame for screen up to validated hits

6 month

Machine/Methods offered under this call

  • FLIPR-Tetra System (Molecular Devices) for fast kinetic imaging by fluorescence or luminescence.
  • 3 ArrayScans (Cellomics Inc./ThermoFisher) one with climate chamber, for sequential detection of up to 6 different fluorescence signal.
  • Impedance/label free analysis (Acea, Excelligence in 96well and in 384well formats) for cellular reporter systems, which can be induced to change ion distribution in cell.
  • FLIPR-Tetra system for kinetic imaging of luminescence or fluorescence readout.

Services provided under this call

  • FLIPR-Tetra System (Molecular Devices) for screening of ion-channel function
  • 3 ArrayScans (Cellomics Inc./ThermoFisher) for all kinds of image-based screening e.g. with luciferase or GFP-tagged proteins and any kind of fluorescence/luminescence.
  • ExCelligence (Acea, impedance/label free measurements)
  • Validation of the assay provided by the user to ensure robustness (all equipment listed above possible) by using 384well format and determination of z-prime factor with positive and negative controls (each 192, half 384well plate)
  • Miniaturization of the assay into 384well plates (all equipment listed above possible)
  • Pilot screen using 5,000 compounds from the EU-OS compound library (all equipment listed above possible)
  • Analysis of pilot screen data (all equipment listed above possible)
  • Full screen with the remaining 95,000 compounds of the EU-OS compound library (all equipment listed above possible)
  • Data analysis and subsequent selection of primary hits (KNIME pipelines already present)
  • Hit picking of primary hits for hit validation (automated Liconic KIWI-store combined with Tecan FreedomEvo workstation)
  • IC50 determination in 9 serial dilutions and duplicate or triplicate measurements of 352 to 704 primary hits selected for hit validation.

We are using KNIME and the software provided by the vendors of microscopes for automated object identificalion and quantification of fluorescent or luminescent detection/quantification of cellular response

Note

Please note that this project includes the possibility of re-screening during potential chemical optimization under the medicinal chemistry call starting in 2021 (tentative date).

Prerequisites for applicants

The applicants needs an assay demonstrating a good signal to noise ratio in your labscale assay, which we will downscale to 20-40 µL volumes in 384well microtiter plates for HTS. Your test needs to pass an assay acceptance test (i.e., half plate negative and the other part positive cellular responses) to validate robustness and not a random instability independent from the target for interference. If your assay demonstrates an appropriate z-prime factor you will be able to start HTS.

As specifics of the assay transfer procedure may vary between partner sites, the applicant and the individual sites will agree on the appropriate steps and logistics together.

 

Cell-based assays using complex cellular systems

Call Overview / Basic Data

Topics / Keywords:

drug discovery, small molecules, high-throughput screening, cell-based assays

Open Time:

Start date:
June-01-2019 (20:00 CET)

Closing date:
September-30-2019 (20:00 CET)

Contacts:

Scientific contact: Jens Peter von Kries, kries@fmp-berlin.de
Technical contact: Martin Neuenschwander, neuenschwander@fmp-berlin.de
Machine/Methods contact: Andreas Oder, oder@fmp-berlin.de; Carola Seyffarth, seyffarth@fmp-berlin.de

Partner site


FVB-FMP

Leibniz Research Institute for Molecular Pharmacology in the Forschungsverbund Berlin e.V.

Screening Unit

Robert-Roessle-Strasse 10

13125 Berlin-Buch

Germany

https://www.leibniz-fmp.de/core-facilities/screening-unit/screening-unit/intro.html