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A FCCS based platform technology accelerates drug discovery

Frank Becker, Intana Bioscience GmbH, Germany

Conducting assays and screens for compound identification in a physiological environment is considered increasingly important. By eliminating hits that score positive only under highly artificial experimental conditions the labor intensive drug discovery process is rationalized and streamlined at an ealy timepoint. Cellular lysates comprise the complete equipment of biomolecules as living cells but uncouple the testing of activity on cellular targets from ADME parameters as cellular uptake and membrane permeability. Thus, cellular lysates bridge the gap between cellular and biochemical enzyme assays concomitant with greatly facilitated assay development and avoids the need of protein purification.
Based on the single molecule sensitive Fluorescence Cross Correlation Spectroscopy (FCCS) we have established a generic, fast and reliable platform technology that enables to assign affinities and rate constants of compound target interactions in cellular lysates. The technology has been successfully employed to test compound-target and protein-protein interactions as well as the binding of nucleic acids to proteins. A library of > 500 human protein kinase domains as GFP fusions has been generated to allow profiling of candidate compounds for yet unknown targets. Moreover, high throughput screenings of compound libraries demonstrated stable assay conditions, reliable readout, great reproducibility and time effective assay development.

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