Beckman Coulter Concept Application (system support)
High-Throughput Screening for Inhibitors of SHP-1 Tyrosine Phosphatase
Paul W. Diaz, Ph.D., Shun Luo, Ph.D., Troy Cook, Graham Threadgill, Ph.D.
and Joe Zhao, Ph.D.
Automated Solutions Development Center, Beckman Coulter, Inc.
Department of Hematology and Oncology, Vanderbilt University
The assay is designed to measure the activity of src-homology2 tyrosine phosphatase (or any tyrosine phosphatase) via the hydrolysis of 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP) to the non-phospho form. In this case the assay is homogeneous, that is to say, there are only additions of reagents to the test well with no washing, withdrawals, filtering etc, prior to reading. The substrate (DiFMUP) is non-fluorescent until hydrolyzed and the assay can be performed in as little as 10 ul in the appropriate plates, such as the Greiner 384 Black Shallow plate etc. The assay itself was optimized using Beckman Coulter Automated Assay Optimization (AAO) version 1.5 and coupled with SAS JMP on the BiomekŪ 2000. The design experiments were then transferred to the Biomek FX and the assay was performed using the Biomek FX.
Features
- Miniaturized fluorescence-based, high-throughput, and homogeneous assay for SHP-1 tyrosine phosphatase
- Execute the screen utilizing a well-defined Coelacanth compound library on the Beckman Coulter SAGIAN CORE system
- Implement an active data management and acquisition system, using FirePower* and SpotfirePro* software, capable of cross platform data integration
- Determine hits and utilize automated methods of compound retrieval and hit confirmation (cherry-picking and dose responses)
* All trademarks are the property of their respective owners. Where applicable, the PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffman-LaRoche, Ltd.
