Obliquely deposited substrates for detection of Ras

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Key cell signaling molecules, such as Ras, play a central role in carcinogenesis and are the targets for a new generation of drugs that inhibit protein kinases. The ability to measure the activation status of molecules along an entire pathway is of increasing importance, yet the available technologies are inadequate to handle the demand for simple assays that permit simultaneous testing of several molecules and can determine their total concentration, activation status, and post- translational modifications, such as phosphorylation or farnesylation. The movement of protein kinase inhibitors and related compounds into clinical trials will increase the demand for such assays in academic, pharmaceutical and clinical laboratories. Platypus technology is a novel, broadly applicable platform nanotechnology based on the sensitivity of liquid crystals to nanoscale features in surfaces. The technology permits highly sensitive detection of molecular interactions and is adaptable to automated and multiplexed formats. In this proposal, we will optimize the technology for the detection and quantification of total Ras and activated Ras, utilizing gold films with anisotropic topography. PROPOSED COMMERCIAL APPLICATIONS: Platypus Technology is a revolutionary platform technology with broad application potential. A rapid method for the analysis of total and activated Ras will have a large market and form the basis of a device to measure multiple molecules along a signaling pathway, determine their amounts and the relative portion of activation.