Screening Strategies in Drug Discovery – Multimedia

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.