Euroscreen FAST is bringing 20 years of expertise and a successful track record for your GPCR needs such as:
- Residence time experiments for Kon/Koff determination
Direct kinetics of ligand-receptor interactions are not usually taken into account in the optimization of a drug’s pharmacokinetics properties. However, several studies have proposed that slow dissociation rates in vitro can be associated to long-acting effects of a compound in vivo.. This important parameter can be measured as the rate at which the complex dissociates (expressed as koff or dissociative half-life) to the free ligand and its target protein. The dissociative half-life has been connected to improved clinical efficacy in a large number of marketed drugs.
Below we exemplify the determination of the kon/Koff for the potent and selective D2L antagonist Risperidone by looking at the association of [3H]-Spiperone in the presence of increasing concentrations of the antagonist
- Cell migration assay (recombinant and primary cells)
Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammation. One such process, leukocyte extravasation, is crucial for appropriate and effective immune response. Leukocytes normally exist in a resting state as they circulate though the body. However, upon interaction with small molecules known as chemoattractants, they rapidly respond with endothelial adhesion followed by emigration from the vasculature and chemotaxis to the site of inflammation.
Below we exemplify the quantitative determination of the CCR5 chemokine MIP-1b as well as potent CCR5 antagonist TAK-779 using recombinant CCR5 CHO-K1 cell line.
-Cytokine Release from LPS stimulated human PBMC
The human LPS-stimulated PBMC model is a good preliminary profiling assay to evaluate a compound's effect on inflammatory cytokine production.
Below we exemplify the inhibitory effect of dexamethasone on pro-inflammatory TNFa cytokine release from LPS-stimulated human Peripheral Blood Mononuclear Cells.
- Rat Isolated b-islet Glucose Stimulated Insulin Secretion (GSIS) assay
Type 2 Diabetes (T2D) is a worldwide public health devastating problem. Drug therapies are available to address this disease, but the common appearance of adverse effects, or lack of efficacy after long-term use, causes concern. Moreover, the growing patient population suffering from T2D and associated metabolic diseases creates a demand for new entrants into this therapeutic market. GPCRs are among the target for the largest number of marketed drug in this field. Taking advantage of our expertise in the field and our extensive catalogue of naturally-coupled GPCR cell-based assays, we are now offering the possibility to support the pre-clinical development and build confidence in the selection of compounds that have desired effects in man.
Below we exemplify the stimulatory effect of GLP-1(7-36) peptide on glucose-induced insulin release from fresh isolated rat islets.
- Glucose Stimulated Insulin Secretion (GSIS)
Fresh isolated Wistar rat islets were incubated with increasing concentration of Glucose with or without GLP-1 for 90 min at 37°C in a 96-well plate. Cell supernatants were harvested and insulin content was measured with a Insulin HTRFTM assay.
- Off-the-shelf and custom-based enzymatic assays
Enzymes represent a fast-growing class of targets in the drug discovery industry, as principal targets for screening like kinases or enzymes implicated in epigenetic modulation, or as potential safety issues when assessing off-target effects. Consequently, we have recently put our scientific expertise to the development and validation of several enzymatic assays including kinases, monoamine oxidases, catechol-O-methyltransferase, phosphodiesterases, cyclooxygenase, or acetylcholine-esterase.
Below we show the Steroid 5-a-Reductase (SRD5a) assay that was developed for a specific application. CHO-K1 cells expressing recombinant SRD5a are used to induce testosterone conversion to androstanolone. This enzymatic activity is dose-dependently inhibited by Linolenic Acid and intact testosterone is quantified using an HTRFTM kit.