Assay Development & Screening - Selvita

[Liz Seyi]

Selvita offers custom assay development and screening/profiling services up to 1536-well plate format. We employ a fully automated HTS platform allowing testing of 10,000 compounds a day across a wide range of biochemical and cellular assay types. We operate HCS screening in support of client projects, inour new state-of-the-art HCS laboratory and are generating novel machine-learning based scripts to support enhanced image analysis in the cell types of particular interest to our clients. To support our activities we have created a robust compound management infrastructure supporting manipulation of samples in racks, tubes and plates.
Our team has an extensive expertise in the areas of molecular biology, biochemistry, immunology and cell biology. We have a successful track record in assay development for various classes of targets, including enzymes (kinases, phosphatases, proteinases, transferases, metabolic enzymes, immuno-modulatory enzymes, DNA/RNA-associated enzymes) and protein receptors (GPCRs, TLRs, RTKs). Selvita’s cell-based assay development service includes number of assays (viability, proliferation, protein biomarkers, intra- and extracellular metabolites, phosphorylation / dephosphorylation, gene and protein expression, signalling pathways, ligand receptors) employing variety of cellular model (cancer, non-cancer, primary, iPSC-derived cells). Together with state-of-the-art equipment allowing a wide variety of detection modes to be implemented, Selvita extensively supports your drug discovery program.
Supported biochemical and cell-based assays with relevant readouts
• Absorbance and Fluorescence
o viability, proliferation (MTS, BrdU)
o metabolites analysis
o ELISA for biomarkers including Luminex multiplexing assays
• Fluorescence Resonance Energy Transfer (FRET)
o proteinase assays
o DNA/RNA-associated enzymes
o protein-ligand interactions
• Time-Resolved FRET (TR FRET)
o kinase/phosphatase assays (HTRF, LanthaScreen)
o ubiquitination assay (HTRF)
o biomarkers analysis (HTRF)
• Fluorescence Polarization (FP):
o kinase/phosphatase assays (Transcreener)
• Luminescence
o viability (CellTiter-Glo)
o kinase/phosphatase assays (ADP-Glo)
o binding and residence time assay (NanoBRET)
o degradation studies, PROTACs (HiBiT)
• Scintillation counting (3H and 14C)
o DNA/RNA-associated enzymes
o analysis of metabolism (e.g. glucose update, lipogenesis)
• Alpha technology bead-based proximity assays:
o biomarkers analysis (AlphaLISA)
o protein-protein interaction (AlphaLISA)
o phosphorylation / dephosphorylation (AlphaScreen)
o multiplexing (AlphaPlex)
Supported biophysical assays
• Surface plasmon resonance (SPR) – determination of binding kinetics, KD, kon/off
• Thermal shift (differential scanning fluorimetry) – protein stability under drug treatment
• Isothermal titration calorimetry (ITC) – analysis of thermodynamic properties of binding
• Fluorescence polarisation (FP) – direct protein-ligand interaction
• Scintillation counting (3H and 14C) – radioligand binding assays
• protein-ligand NMR
Selvita can 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).
Our in-house capabilities for reagent generation include in-house production and validation of recombinant cell lines and recombinant proteins.
Our assay development & screening strategies are characterized by efficient and effective protocols. Usually, multi-factor design of experiments is employed during method development.  It accelerates optimization of the assay performance, thus helping to obtain a measurement system that meets its requirements. Statistical parameters are used to support a reliable and robust assay for routine use, e.g. IC50/EC50; mRSD (mean Relative Standard Deviation); MSR, S/N ratio; Z’ factor; plate uniformity (read more here).