Roche's XCELLigence System Offers Dynamic And Label-free Assessing Of Kinase Activity In Living Cells

�The central role of receptor tyrosine kinases (RTKs) in cellular processes, especially in cancer, has made them an important prey for several antibody- and small molecule-based inhibitors specific for diverse RTKs for the treatment of dissimilar tumour entities. The electrical impedance detection method with Roche�s real time cell microelectronic sensor-based political program xCELLigence system is a facile, high gear content, and cell based kinase assay allowing for monitoring forgetful term RTK activation and the long term biologic effect of the activation in substantial time in a undivided well. The novel technology has improved utility all over existing in vitro and cell based assay in the identification and characterisation of selective and potent kinase inhibitors, as shown in a recent report.


Several approaches have been developed to identify RTK inhibitors such as antibody-dependent and independent technologies. However, most of these stream approaches ar end point, in vitro based assays that require substantial reagent optimization, and are inadequate in providing information on their effective intracellular activeness. Roche Applied Science's xCELLigence System - originally invented by the US-based ACEA Biosciences and co-developed by Roche and ACEA - addresses several of these limitations. Unlike other RTK assays, this technology is cell-based, label-free, capable of monitoring cellular changes in real-time, and non-invasive. The method utilizes an electronic readout of impedance to quantify cellular status, including cell number, viability, morphology, and cytoskeletal dynamics. Cells are seeded in E-Plate microtiter plates, which are integrated with microelectronic detector arrays. The interaction of cells with the microelectrode surface leads to the generation of a cell-electrode impedance reply, which indicates the position of the cells in terms of morphology, lineament of adhesion and number.


According to experimental information, the xCELLigence System provides a facile, easy platform for identification and further characterization of RTK inhibitors. In a recent study1, the energising of epidermic growth factor (EGF)-mediated changes of COS7 cells pretreated with epidermal growth factor receptor (EGFR) inhibitor and insulin were monitored exploitation the xCELLigence System.


As the authors concluded, the electrical electric resistance detection method was able to blind, identify, and characterize a potent and selective EGF receptor inhibitor from a compound library. The assay quantified morphologic changes in response to growth constituent treatment and therefore mimics proximal events in kinase activation. Additionally, the xCELLigence System provided valuable information about the state of the cell and the signalling pathways being activated. Furthermore, the technology does not take intensive optimisation or special reagents such as peptides, antibodies, or probes, nor suffers from assay constituent interference. Since the readout is noninvasive, multiple treatments can be performed in the same well. The assay pot also be used in conjunction with other existing cell-based assays for RTK. More important, because the assay is cell based, the studies are done in a physiologically relevant environment, allowing for simultaneous assessment of a compound's solubility, stability, membrane permeability, cytotoxicity, and off-target interaction effects. Finally, the system requires selfsame little user training, devising this check amenable for use in both primary and secondary screens.


For more information on the technology, please visit wWW.xcelligence.roche.com


1 xCELLigence System, Application Note No. 4, 2008, Roche Applied Science.

About Roche


Headquartered in Basel, Switzerland, Roche is one of the world's leading research-focused healthcare groups in the fields of pharmaceuticals and diagnostics. As the world's biggest biotech company and an groundbreaker of products and services for the early detection, prevention, diagnosing and handling of diseases, the Group contributes on a broad range of fronts to improving people's health and quality of life. Roche is the world drawing card in ex vivo diagnostics and drugs for cancer and transplantation, and is a market leader in virology. It is also active in former major sanative areas such as autoimmune diseases, inflammatory and metabolic disorders and diseases of the central nervous system. In 2007 sales by the Pharmaceuticals Division totalled 36.8 billion Swiss francs, and the Diagnostics Division posted sales of 9.3 billion francs. Roche has R&D agreements and strategical alliances with numerous partners, including bulk ownership interests in Genentech and Chugai, and invested over 8 billion Swiss francs in R&D in 2007. Worldwide, the Group employs about 80,000 people. Additional information is available on the Internet at hypertext transfer protocol://www.roche.com.


XCELLIGENCE is a trademark of Roche.


E-PLATE, RT-CES, and ACEA BIOSCIENCES are registered trademarks of ACEA Biosciences, Inc. in the US.


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