Transnational Call

SMALL MOLECULE SCREENING

 

Screening for enzyme inhibition and protein binding

Abstract

IMG offers high-throughput compound profiling with cell-based, enzymatic activity and binding assays using a wide range of luminescent or fluorescent readouts such as fluorescence intensity (FI), Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) including HTRF, LanthaScreen, LANCE, and TransScreener; polarized fluorescence (FP), luminescence and BRET in 384/1536-well format.

IMG provides complex services to biochemists and biologists from standard assays to the development of new assays and analysis of the results.

The Institute of Molecular Genetics of the ASCR, v. v. i., is a node of the Czech screening initiative CZ-OPENSCREEN (https://www.openscreen.cz/en/node/106).

Description of technology offered under this call

CZ-OPENSCREEN provides complex services to biologists from standard assays to the development of new assays and analysis of the results. For chemists, CZ-OPENSCREEN offers compound profiling of small molecule libraries using a panel of cell lines and data analysis. CZ-OPENSCREEN develops dedicated software tools, which are available for free. The infrastructure has a fully-equipped biochemistry and molecular biology laboratory for the development of new assays and methods and three fully equipped cell culture laboratories.

CZ-OPENSCREEN combines approaches using cell-based, enzymatic activity and binding assays for high-throughput profiling of compound libraries. Signal detection relies on a wide range of luminescent or fluorescent readouts such as fluorescence intensity (FI), Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) including HTRF, LanthaScreen, LANCE, and TransScreener; polarized fluorescence (FP), luminescence and BRET in 384/1536-well format.

Number of compounds screened

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

Time frame for screen up to validated hits

4 months

Machine/Methods offered under this call

Integrated robotic HTS station for screening of chemical compounds (cell::explorer - PerkinElmer) consists of 6-axis robotic arm (Denso), liquid handling station Janus Mini (PerkinElmer) with pipetting head 384 and Pin Tool (V&P Scientific), multimode reader EnVision (PerkinElmer), HCS microscope Operetta (PerkinElmer), 2 integrated CO2 incubators STX110 with carousels for 110 plates (Liconic), centrifuge Vspin (Agilent) and plate washer ELX405 (Biotek), liquid dispenser MultiDrop Combi (Thermo), non-contact acoustic compound dispenser Echo 550 (Labcyte), plate shaker Variomag (Thermo), barcode reader, high capacity plate stacker and plate sealer PlateLoc (Agilent).

Integrated system for compound storage and sample preparation consists of several interconnected modules: two automated and integrated freezers ASM store (Hamilton) (-20°C) with inert environment (N2) for long-term storage of chemical compounds and logistics module ASMserver (Hamilton). These 3 modules are connected to the integrated robotic liquid handling workstation Vantage (Hamilton), which is used for dispensing, copying and reformatting of microplates in 96/384/1536 formats.

Services provided under this call

  • Consultation on the assay development with respect to its adaptation for High-Throughput-Screening (HTS) and building of the counter-screen assays to eliminate non-specifically acting compounds. Detection of activity is typically carried out spectrophotometrically by multimode plate reader Envision (PerkinElmer).
  • Validation of the assay provided by user, miniaturization for 384/1536-well plate format and adaptation for the screening campaign.
  • Pilot screen with 5,000 compounds, data analysis and further assay development if needed.
  • High-throughput screening (HTS) in 384/1536-well plate format.
  • Data processing, normalization, analysis and hit identification (up to 1% of total screened compounds) using in-house developed Laboratory Information Management System (LIMS).
  • Dose-response testing of primary hits to confirm their activity (6 concentration points)
  • Data processing, normalization, analysis and identification of confirmed hits.
  • Dose-response profiling of confirmed hits with primary, orthogonal (if available) and counter-screen assays to validate their activity (8 concentration points, triplicates).
  • Data processing, normalization, calculation of EC/IC50 and analysis.
  • Interpretation of results and reporting.
  • Cheminformatics support (structure-based clustering of active/non-active compounds, building of preliminary SAR model, data mining) with Chemaxon software, RDkit etc.

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

Potential users are expected to have a mature, robust assay, which can be further miniaturized and adapted for automation. Users must be able to provide purified proteins (e.g. enzymes) with suitable activity, purity and stability and at sufficient amounts (commercially available or user-made) for primary HTS screening campaign (100.000 compounds) and for confirmation/validation assays.

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.

 

Biochemical / Biophysical assays

Call Overview / Basic Data

Topics / Keywords:

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

Open Time:

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

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

Contacts:

Scientific contacts: Petr Bartůněk, bartunek@img.cas.cz, David Sedlák sedlak@img.cas.cz

Technical/Machine/Methods contact: David Sedlák, sedlak@img.cas.cz

Partner site


IMG

Institute of Molecular Genetics of the ASCR, v. v. i.,

CZ-OPENSCREEN: National Infrastructure for Chemical Biology,

Vídeňská 1083,

142 20 Prague 4, Czech Republic

https://www.img.cas.cz/research/petr-bartunek/

https://openscreen.cz/en