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SpeedScreen: label-free liquid chromatography-mass spectrometry-based high-throughput screening for the discovery of orphan protein ligands.

Muckenschnabel, Ingo and Falchetto, Rocco Angelo and Mayr, Lorenz and Filipuzzi, Ireos (2004) SpeedScreen: label-free liquid chromatography-mass spectrometry-based high-throughput screening for the discovery of orphan protein ligands. Analytical Biochemistry, 324 (2). pp. 241-249. ISSN 0003-2697

Abstract

A high-throughput screening methodology tailored to the discovery of ligands for known and orphan proteins is presented. With this method, labeling of neither target protein nor screened compounds is required, as the ligands are affinity selected by incubation of the protein with mixtures of compounds in aqueous binding buffer. Unbound small-molecular-weight compounds are removed from the target protein:ligand complex by rapid size-exclusion chromatography in the 96-well format. The protein fraction is analyzed subsequently by liquid chromatography-mass spectrometry for detection and identification of the bound ligand. This screening method was validated with known protein:ligand model systems and optimized for selection of high-affinity binders in an industrial screening environment. All sample handling steps and the analytics are rapid, robust, and largely automated, adopting this technology to the needs of present high-throughput screening processes. This affinity-selection technology, termed SpeedScreen, is currently an integral part of our lead discovery process.

Item Type: Article
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Additional Information: author can archive post-print (ie final draft post-refereeing); Publisher's version/PDF cannot be used
Keywords: Drug discovery; Lead finding; High-throughput screening; Deconvolution of mixtures; Affinity selection; Orphan proteins
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Date Deposited: 14 Dec 2009 13:54
Last Modified: 31 Jan 2013 01:06
URI: https://oak.novartis.com/id/eprint/783

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