Abstract

A novel biosensing platform was developed by integrating an aptamer-based DNA biosensor with graphene oxide (GO) for rapid and facile detection of adenosine triphosphate (ATP, as a model target). The DNA biosensor, which is locked by GO, is designed to contain two sensing modules that include recognition site for ATP and self-replication track that yields the nicking domain for Nt.BbvCI. By taking advantage of the different binding affinity of single-stranded DNA, double-stranded DNA and aptamer-target complex toward GO, the DNA biosensor could be efficiently released from GO in the presence of target with the help of a complementary DNA strand (CPDNA) that partially hybridizes to the DNA biosensor. Then, the polymerization/nicking enzyme synergetic isothermal amplification could be triggered, leading to the synthesis of massive DNA amplicons, thus achieving an enhanced sensitivity with a wide linear dynamic response range of four orders of magnitude and good selectivity. This biosensing strategy expands the applications of GO-DNA nanobiointerfaces in biological sensing, showing great potential in fundamental research and biomedical diagnosis.

Highlights

  • Due to the obvious advantages, such as highly specific Watson–Crick base pairing interactions, and single-stranded flexibility, DNA is widely employed as a smart building block in the design of DNA-based biosensors [1,2,3,4,5]

  • The design of the DNA biosensor and the operation mechanism of the amplification system are illustrated in Figure 1, in which ATP is adopted as the model target

  • The measured data offer unmistakable evidence that the aptameric DNA biosensor coupled with polymerase/nicking enzyme synergetic isothermal amplification capability can be employed in graphene oxide (GO)-based sensing platform

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Summary

A Graphene-Based Biosensing Platform Based on Regulated

The Ministry-Province Jointly Constructed Base for State Key Lab-Shenzhen Key Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China.

Introduction
Materials and Reagents
Apparatus
Detection of ATP by Using the Biosensing Platform
Selectivity Investigation of the Proposed Biosensing Platform
Results and Discussion
Feasibility of the Biosensing Platform for ATP Detection
Absorption and Regulated Release of the DNA Biosensor
Quantitative Detection of ATP
Selective Measurement of ATP
Application of the Biosensing Platform in Real Sample
Conclusions
Full Text
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