47 Rue Adrienne Bolland
To detect the target biomarker for your drug discovery projects in human iPSC-derived cell cultures, Ncardia develops and executes customized AlphaLISA assays. This technology allows for the sensitive detection of molecules of interest with a wide dynamic range and reproducible performance. Since the assay is homogeneous, biological interactions that would normally be disrupted by washing or separation steps are preserved. This allows for more physiologically relevant results and makes the assay compatible with high-throughput screening.
Hypertrophic cardiomyopathy (HCM) is an autosomal dominant disease of the cardiac sarcomere, resulting in elevated risk for clinical complications. The natriuretic peptide fragment NT-proBNP is an established clinical biomarker for the diagnosis of hypertrophy. Ncardia has developed a specialized NT-proBNP AlphaLISA assay to detect the secretion of this biomarker in cell culture supernatants from in vitro HCM models.
In this case study, a primary high throughput compound screen was performed, consisting of 3830 compounds in fifteen 384-well microplates. Following a primary screen, a hit confirmation analysis was performed for 341 compounds, using three distinct assays: NT-proBNP secretion AlphaLISA (technical duplicate), the AlphaScreen TruHits assay to deselect false positives, and high content imaging (HCI) to assess intracellular proBNP expression.
Primary hits were first confirmed by performing the NT-proBNP AlphaLISA at a 1 μM concentration for each compound. Compounds with percent inhibition (PIN) >40 % (red line) were confirmed as hits (A). TruHits assays were then performed to deselect false positives i.e. quenchers, scatterers and biotin mimetics (B). As a third step, the NT-proBNP AlphaLISA was multiplexed with HCI to confirm intracellular proBNP expression (C.). Red dots in the graph represent the MAX effect reduction in proBNP levels, and blue dots represent the MIN effect.
Click here to read the full case study: <Link to be added when Tecan whitepaper is approved>.
Would you like to know more about our biomarker detection service?
We're here to listen to your project plans, questions and challenges, and to co-develop a solution that fits your specifications. Please leave your details and message in the form below and we will get back to you as soon as possible.