No abstract

We have developed a bound/free separation system using a vascular endothelial growth factor (VEGF) aptamer and a peptide nucleic acid (PNA) to detect VEGF. In this system, we designed capture PNA (CaPNA), which hybridizes with the aptamer in the absence of the target protein, but does not hybridize with the aptamer in the presence of the target protein due to steric hindrance and/or stabilization of the aptamer’s structure. By removing the aptamers not bound to the target protein using CaPNA immobilized beads, we can detect the target protein by measuring signals labeled with the aptamer in the supernatant. In this study, we detected VEGF using CaPNA-immobilized beads without the time-consuming washing step. This simple and rapid system can detect 25 nM of VEGF in 15 min.

We propose a novel enzyme-labeling method for DNA aptamers using enzyme-fused zinc finger proteins. We achieved thrombin detection and vascular endothelial growth factor detection using zinc finger-fused firefly luciferase.