Assay development services

In vitro pharmacodynamic (PD) models and assays are used to obtain useful quantitative information on the effect of either single drugs or drug combinations against the disease of interest.

Cell and Molecular Biology Laboratory at Selvita offers development, optimization and validation of a broad range of cell-based functional assays, as well as bioanalytical and biochemical tests in the area of oncology, CNS, autoimmune disorders and other disease areas.

Those techniques constitute an integral part of lead identification and optimization as an important part of the drug discovery and development process.

In vitro oncology platform

  • Selection and validation of in vitro cellular disease models

  • Validation of target proteins in functional cellular assays

  • Established viability/proliferation assays

  • Cell cycle analysis

  • Established apoptosis/cell death assays

  • LDH (Lactate dehydrogenase activity) – cell death assay (screening format, dose-response curves)

  • FACS early apoptosis detection assay (Annexin V/PI staining)

  • PARP cleavage for detection of apoptosis

  • PARP activity colorimetric assays

  • Caspase activity – apoptosis assay (labeled substrates)

  • Clonogenic survival assay

  • Senescence induction assay (in cells β-Galactosidase activity assay) 

  • Combination index for evaluation of synergy

  • Cell signaling profiling

In vitro CNS platform

  • Monitoring of cells in response to the effects of test compounds on GPCRs

  • In-house generation and validation of cloned receptors cell lines

  • cAMP production/inhibition (screening format, dose-response curves)

  • Expertise in handling compounds with low solubility in water and/or high lipophilicity

In vitro inflammatory and autoimmune diseases platform

  • In vitro inflammasome activity assays

  • Caspase 1 activity assay

  • Production of interleukin 1β and 18

  • Cell-based assays specific for inflammation

  • Activation of signal transduction by ligands of Toll-like receptors and NOD-like receptors

  • Production of pro-inflammatory cytokines

  • Key proteins phosphorylation during signal transduction

  • Activation of inflammatory caspases

Other assays

  • Cell metabolism – lipogenesis, lipolysis, glucose uptake etc.

  • Cell death, proliferation and differentiation

  • Cell migration, invasion and chemotaxis assays

  • Cell activation and downstream events characterization

  • Ligand binding, receptor activation, receptor binding

  • Assays specific for monoclonal antibodies – neutralization assays, antibody-dependent cell-mediated cytotoxicity etc.

  • Enzymatic assays, protein-protein and drug-protein interaction assays

During assay development we offer

  • Case-by-case selection of suitable cellular model/s to investigate the biological mechanism of interest (e.g. cancer cell panels of a specific type and/or with common mutation patterns)

  • Routine examination of cell line for mycoplasma contamination prior to the assay development

  • In-house production of siRNA knocked down cell lines as models lacking protein of interest (if needed)

  • Experiments in suitable cell culture conditions (culture medium supplementation, incubation parameters – humidity, temperature, O₂/CO₂ concentration)

  • HTS analysis of large compound libraries with the use of Hamilton STARlet with iSWAP Plate Handler combined with the multimode plate reader

  • Various detection modes – absorbance, fluorescence intensity, fluorescence polarization, time-resolved fluorescence (TRF) – LANCE and DELFIA technologies, luminescence measurements, AlphaLISA and AlphaScreen technologies

  • Assay validation with the use of reference and control compounds/proteins

  • Full validation of cell-based assay and bioassay in terms of linearity, precision, accuracy as well as inter- and intra-laboratory variability

  • Bioassay validation according to EMA and FDA guidelines

  • GLP/GMP-compliant assay development and validation