Turn-On Aptameric System for Simple and Selective Detection of Protein via Base Stacking-Dependent DNA Hybridization Event

Abstract

Base stacking is employed in an entirely new type of sensing platform for the simple and robust detection of protein. Only in the presence of protein, the aptamer DNA can hybridize stably with the capture DNA to form a stem-loop structure due to the enhancement of base stacking. This leads to a strong chemiluminescence emission for simple protein detection. With the use of a platelet-derived growth factor as a model, a fM detection limit was obtained with a dynamic range that spanned 4 orders of magnitude. Upon modification, the approach presented herein was also extended to detect other types of targets including Hg(2+) ion and adenosine and also other types of labels such as fluorescence nanogold. We believe such advancements will represent a significant step toward improved diagnostics and more personalized medical treatment and environmental monitoring.

Ultrasensitive Fluorescence Polarization Aptasensors Based on Exonuclease Signal Amplification and Polystyrene Nanoparticle Amplification

“…Scheme depicts the third configuration of an amplified FP aptasensor for the detection of platelet‐derived growth factor BB (PDGF‐BB). This sensing platform involves the use of a target‐triggered, base stacking‐dependent DNA hybridization method 20. An anti‐PDGF‐BB aptamer probe was designed with four base pairings and sixteen bases at its extension 5′‐end, and a DNA‐2 probe modified at its 5′‐end with FAM and at its 3′‐end with PS NPs was designed to be partly complementary to the extension sequence of the aptamer.…”

“…These two probes coexist stably in the absence of PDGF‐BB, and the FAM dye has a very high P value. In the presence of PDGF‐BB, the anti‐PDGF‐BB aptamer probe binds to PDGF‐BB and forms a stem‐loop structure, thereby leading to an enhancement of base stacking 20. This promotes the hybridization of the PDGF‐BB–aptamer complex with the FAM‐labeled DNA‐2 probe, which triggers the partial enzymatic cleavage of the FAM‐labeled DNA‐2 probe by T7 Exo, thus resulting in the removal of FAM dye from PS NPs, the release of the PDGF‐BB–aptamer complex, and a decrease in the P value.…”

Microplate Chemiluminescent Assay for DNA Detection Using Apoperoxidase-Oligonucleotide as Capture Conjugate and HRP-Streptavidin Signaling System

“…The methods of DNA detection are commonly based on a hybridization reaction [ 6 , 7 , 8 , 9 ]. These methods can be divided on homogeneous and heterogeneous ones.…”

“…The optical methods of DNA detection are usually developed with the use of gold or carbon nanoparticles (NPs) [ 16 , 17 , 18 , 19 ] and magnetic beads [ 20 , 21 ]. An alternative solid carrier for DNA detection are microtiter plates [ 9 , 22 , 23 ], which are widely applied in the production of ELISA kits.…”

“…Covalent binding of DNA oligonucleotides on the microplate surface is also used. Presently the microtiter plates with different activated groups are commercially available and can be used for immobilization of amino-containing oligonucleotides [ 9 , 31 ]. The main drawback of such plates is their high cost.…”