UiB is a specialist screening site, integrated together with the Biophysical and Structural Biology facilities in the BiSS Platform (http://www.uib.no/en/rg/biss). UiB offers a molecular/biophysical-based screen, monitored by differential scanning fluorimetry (DSF) searching for stabilizer compounds that can be developed into inhibitors or pharmacological chaperones for the correction of misfolding genetic disorders.
The screening site SC-NO University of Bergen (UiB) is a node of the Norwegian screening initiative NOR-OPENSCREEN (http://www.openscreen.no/). The research group Biorecognition uses a combination of computational and experimental techniques, within the fields of biophysics, biochemistry and cellular biology. UiB has a particular focus on drug discovery for conformational genetic diseases and neurotransmitter disorders and new antibiotics. Collectively, the group has extensive experience with high-throughput screening (HTS) based on differential scanning fluorimetry (DSF) and virtual screening.
DSF constitutes a sensitive assay of ligand binding and an efficient biophysical readout for target-based screening. Customarily, DSF is performed in a real-time PCR instrument with 384- well microplates with the purified target protein sample at concentrations in the range 0.05–0.15 mg/mL in an optimal buffer, and the dye SYPRO Orange that emits fluorescence when interacting with hydrophobic areas of denatured proteins.
The compounds are added to the assay solution to a final concentration of 80 μg/mL and 4% DMSO. Negative DMSO controls are routinely included on each plate. The unfolding curves are then registered from 20 to 95 °C at a 2 °C/min scan rate, and the midpoint melting temperature (Tm) and the corresponding shift relative to the DMSO reference (ΔTm) are calculated for each compound using in-house software. Compounds with significant increases in ΔTm are selected and those showing adequate follow-up concentration-dependent DSF curves are provided as primary stabilizing hits.
100,000 (full commercial set) of the EU-OPENSCREEN ERIC compound library
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).
The prerequisite starting point to qualify for access is the availability of a purified protein target (approximately 100 mg for screening of the 100.000 compounds). UiB is not locked to specific target classes or disease areas. The applicant should have characterized the stability of the target, performed buffer screens for selection of optimal buffer, characterization of stability to freezing and thawing and best storage conditions for the protein of interest.
The applicant should have a functional assay that allows validation of hits and discrimination of the stabilizing ligands in possible inhibitors, activators or just stabilizers without effect on the activity of the target.
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
drug discovery, small molecules, high-throughput screening, biochemical/biophysical assays
June-01-2019 (20:00 CET)
September-30-2019 (20:00 CET)
Scientific contact: Aurora.firstname.lastname@example.org
Technical / machine / methods contact: Emil.email@example.com