The use of proteomics for surrogate markers of prion infection

We propose new methods for discovery of TSE biomarkers based on combing the MultiPhoton Detection (MPD) technique with methods of proteomics, especially by applying MPD enhanced differential display of proteins (dd-PROT/MPD). This MPD enabled techniques, developed by the group of Dr. A. K. Drukier, enables more sensitive and less expensive tests for correlating the levels of low abundance protein ("molecular switch") with TSE infections. These differentially displayed proteins will then be validated as TSE biomarkers. The primary objective of Phase I of this proposal is to discover differentially displayed proteins associated with TSE. We will further develop methodology for the Multi Photon Detection (MPD)-enhanced differential display of proteins (dd-PROT/MPD) and apply it to determining proteomic differences in the CSF, brain tissue and blood of healthy and prion infected hamsters. Since the quantitative characterization by 2D-PAGE with the MPD Imager permits the display of proteins with about a 100-fold better sensitivity than current methods (spots with 50 zeptomole of protein can be detected), rare proteins that are expressed differentially in the disease state can be reliably identified. Hopefully, some of them will also be present in blood to allow ante mortem TSE diagnosis.