Bioprofiling Assays

Description

All compounds in the EU-OPENSCREEN compound collection will be characterised for a basic set of properties which are relevant for chemical biology or drug discovery projects. Compounds entering the EU-OPENSCREEN compound collection will be carefully characterised and annotated for basic physicochemical (e.g. solubility, compund aggregation) and biological properties (e.g. cell viability, anti-microbial activity) by testing them in a standard panel termed ‘bioprofiling’ assays´. Bioprofiling of the EU-OPENSCREEN compound collection in a standardised manner will contribute to the wealth of quantitative data on compound bioactivities and will comprehensively characterise each of the compounds before they are utilised by the scientific community. Bioprofiling services will be provided at one or more partner sites in the EU OPENSCREEN ERIC network. Lot 1: Aggregation Physicochemical Assay 1: The formation of soluble compound aggregates is a major cause of false positives and distorted values in HTS assays. Lot 2: Solubility Physicochemical Assay 2: Poor solubility can mask compound activity in bioassays inkluding underestimated activity, reduced hit rates in HTS and variable data outputs. Lot 3: Determination of pKa Physicochemical Assay 3: The ionisation value of a compound reflects the diffusion rate across membranes and influences lipophilicity, solubility and protein binding, and therefore is an important parameter for compound optimization. Lot 4: Determination of logP (partition coefficient) and logD (distribution coefficient) Physicochemical Assay 4: These parameters describe the lipophilicity of neutral (logP) and ionizable compounds (logD) and are therefore important values for predicting their behaviour (solubility, permeability) at physiological environments and different pH. Lot 5: Interference with common Bioluminescence reporters Biochemical Assay 5: Important readout for many cellular reporter format assays. Will identify potential screening artefacts/false positives based on modulation of reporter enzymes. Lot 6: ROS (Reactive Oxygen Species) Biochemical Assay 6: Redox active compounds are commonly found in many screens as false positives as they can oxidize disulfide bridges in proteins or react with important co-factors. Lot 7: Cell viability Cell-based Assay 7: Detection of the number of living cells by various means, e.g. measurement of ATP. Lot 8: Antibacterial and antifungal assays Biological Assay 8: Deliver information about potential antibacterial and antifungal properies by testing compounds in a panel of growth assays. Lot 9: Cell-painting Cell-based Assay 9: Multiplexes many readouts based on the use of several fluorescent dyes to reveal broadly relevant changes of cellular pomponents or organelles.