High-Throughput Screening – Multimedia

In this study, we have used an Acumen Explorer equipped with a violet 405nm laser in conjunction with a selection of shorter-wavelength amine-reactive fluorophores (Invitrogen, Molecular Probes) to demonstrate a greater utility for blue fluorescent probes within high content assays.

The DiscoveRx ADP Hunter™ assay measures the generation of ADP resulting from kinase phosphorylation of a substrate, in a format specifically designed for the high throughput screening laboratory. ADP is measured through a coupled enzyme system, which is linked to a positive, red-shifted fluorescent readout. The assay has a wide dynamic range (1.2 – 120 µM), and can be used with a wide range of ATP, from less than 5 µM up to 300 µM.

Here we present a web-based bioinformatics solution for the management and analysis of all areas of cell-based screen experimentation: the Laboratory Information Management System of the Technology Development Studio (TDS-LIMS). TDS-LIMS is a software package consisting of the Library Checker, Library Browser, Scheduling Kit, Screen Browser and Screen Publisher.

The aim of this project was to develop high throughput screening methods using an IMSTAR Pathfinder™ High-content imaging platform to support the analyses of polymeric materials synthesised using a parallel approach. This was achieved by developing arrays of polymeric materials.

High-density screening platforms are an integral part of any successful high-throughput screening facility. The reduced assay volumes associated with increasing well density lowers reagent costs, accelerates assay throughput, and can improve assay quality and reproducibility by minimizing assay duration.

To build a unique kinase platform providing reliable and quantitative data, we have designed a flexible system with well-defined yet different assay conditions for each kinase. This platform minimizes interferences, including those from test compounds, and displays a homogeneous and generic assay format.

Here, we model the integration of the Protedyne BioCube System and the Acumen Explorer for practical high-throughput, high content screening of protein kinases.

In this study, using Protein Kinase A (PKA) as are presentative assay system, peptide phosphorylation reactions carried out in 384 PH filter plates showed dependence on enzyme and substrate concentrations, and on time. Kinase inhibition studies demonstrated IC50 and Z’ values equivalent to those obtained in 96-well plates. The higher throughput 384 well format offers a miniaturized version of the 96 well assay with equivalent sensitivity and reproducibility.

Using the Muscarinic M1 G-protein Coupled Receptor as a model system, we achieved accurate and reproducible determinations of binding affinity (Kd) and IC50 results for known ligands on the 384-well plate device. As a result of being able to incubate the reaction mixture in the filter plate and configure the assay using half the reaction volume, significant reductions in reagent costs and radioactive waste were achieved.