Sensitive Discrimination and Detection of Prion Disease-Associated Isoform with a Dual-Aptamer Strategy by Developing a Sandwich Structure of Magnetic Microparticles and Quantum Dots

The major challenge of prion disease diagnosis at the presymptomatic stage is how to sensitively or selectively discriminate and detect the minute quantity of disease associated prion protein isoform (PrPRes) in complex biological systems such as serum and brain homogenate In this contribution, we developed a dual-aptamer strategy by taking the advantages of aptamers, the excellent separation ability of magnetic microparticles (MMPs), and the high fluorescence emission features of quantum dots (QDs) Two aptamers (Apt1 and Apt2), which can recognize their two corresponding distinct epitopes of prion proteins (PrP), were coupled to the surfaces of MMPs and QDs, respectively, to make MMPs Apt1 and QDs Apt2 ready at first, which then could be coassociated together through the specific recognitions of the two aptamers with their two corresponding distinct epitopes of PrP, forming a sandwich structure of MMPs-Aptl-PrP Apt2-QDs and displaying the strong fluorescence of QDs Owing to the different binding affinities of the two aptamers with PrPRes and cellular prion protein (PrPC), both of which have distinct denaturing detergent resistance, our dual-aptamer strategy could be applied to discriminate prp(Res) and PrPC successfully in serum Further identifications showed that the present dual aptamer assay could be successfully applied to the detection of PrP in 0 01% brain homogenate, about 1000 fold lower than that of commonly applied antibody-mediated assays, which can detect PrP just in 10% brain homogenate, indicating that the present designed dual-aptamer assay is highly sensitive and adequate for clinical diagnosis without isolation of target protein prior to assay