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AlphaScreen™ Kinase HTS Platforms

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Kinases represents one of the most important family of targets in high throughput drug screening. Tyrosine kinases and serine / threonine kinases are known to play key roles in signal transduction as well as in cell growth and differentiation. Intense screening campaigns are underway in all major pharmaceuticals and large biotech companies to find kinase inhibitors for the treatment of inflammatory diseases, immunological disorders and cancer. The present contribution describes models that were developed to produce kinase assays amenable to HTS using AlphaScreen. Because of the flexibility allowed by AlphaScreen, kinase assays can be developed using direct or indirect approaches. Tyrosine kinase assays are usually performed with a direct format involving generic anti-phosphotyrosine antibodies while serine / threonine kinase assays are performed with an indirect format where specific antibodies are captured using protein A conjugated Acceptor beads. Streptavidin-coated Donor beads are used to capture either generic (ex. poly GT) or specific biotinylated substrates. Herein, are presented different methods to perform screening for inhibitors acting on the soluble β- insulin receptor tyrosine kinase (IRKD), and on p38, a member of the MAP kinase family.

Keywords: AlphaScreen; Tyrosine kinases; protein

Document Type: Review Article


Affiliations: PerkinElmer Life and Analytical Sciences, PerkinElmer-BioSignal Inc., Montreal, Canada.

Publication date: 2004-03-01

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