A Facile, Label-Free, and Universal Biosensor Platform Based on Target-Induced Graphene Oxide Constrained DNA Dissociation Coupling with Improved Strand Displacement Amplification.

In this work, we report a low-cost and easy operation biosensor platform capable of detection of various analytes with high sensitivity and good selectivity. By ingeniously assigning the specific aptamer into a primer-template integrated DNA template, and using monolayer graphene oxide as a reversible and nonspecific inhibitor, the simple biosensor platform is set up. Without a target, the DNA template is constrained by the graphene oxide sheet and results in low signal. In the presence of a target, the constrained DNA template is released from the graphene oxide surface via a target-induced aptamer conformational change, and further amplified through the improved strand displacement amplification reaction. Therefore, the target detection is simply converted to DNA detection, and a correlation between target concentration and fluorescence signal can be set up. As a result, dozens-fold signal enhancement, high sensitivity, good selectivity, and potential practicability are achieved in target detection. More importantly, the proposed biosensor platform is versatile, meaning that it can greatly facilitate the detection of a variety of analytes. Due to the low cost and easy availability of sensing materials, and the elimination of tedious detection operations, we believe that this simple and universal biosensor platform can find wide applications in biological assay and environment monitoring.

[1]  B. Wang,et al.  Graphene‐Oxide‐Conjugated Polymer Hybrid Materials for Calmodulin Sensing by Using FRET Strategy , 2015 .

[2]  Qian Wang,et al.  Graphene fluorescence switch-based cooperative amplification: a sensitive and accurate method to detection microRNA. , 2014, Analytical chemistry.

[3]  Kun Wang,et al.  Multiple signal-amplification via Ag and TiO2 decorated 3D nitrogen doped graphene hydrogel for fabricating sensitive label-free photoelectrochemical thrombin aptasensor. , 2018, Biosensors & bioelectronics.

[4]  M. Wagner.  Single-cell ecophysiology of microbes as revealed by Raman microspectroscopy or secondary ion mass spectrometry imaging. , 2009, Annual review of microbiology.

[5]  Muling Shi,et al.  Aptasensor with Expanded Nucleotide Using DNA Nanotetrahedra for Electrochemical Detection of Cancerous Exosomes. , 2017, ACS nano.

[6]  Jing Zhang,et al.  Highly sensitive multiple microRNA detection based on fluorescence quenching of graphene oxide and isothermal strand-displacement polymerase reaction. , 2012, Analytical chemistry.

[7]  Modi Wang,et al.  Development of an Iridium(III) Complex as a G-Quadruplex Probe and Its Application for the G-Quadruplex-Based Luminescent Detection of Picomolar Insulin. , 2016, Analytical chemistry.

[8]  J. Justin Gooding,et al.  Proximity extension of circular DNA aptamers with real-time protein detection , 2005, Nucleic acids research.

[9]  Zhengping Li,et al.  Precise Quantitation of MicroRNA in a Single Cell with Droplet Digital PCR Based on Ligation Reaction. , 2016, Analytical chemistry.

[10]  Eun Jeong Cho,et al.  Real-time rolling circle amplification for protein detection. , 2007, Analytical chemistry.

[11]  B. Spengler,et al.  Single cell matrix-assisted laser desorption/ionization mass spectrometry imaging. , 2012, Analytical chemistry.

[12]  Jian-hui Jiang,et al.  Imaging Endogenous Metal Ions in Living Cells Using a DNAzyme-Catalytic Hairpin Assembly Probe. , 2017, Angewandte Chemie.

[13]  Yixiang Duan,et al.  Magnified fluorescence detection of silver(I) ion in aqueous solutions by using nano-graphite-DNA hybrid and DNase I. , 2014, Biosensors & bioelectronics.

[14]  C. Litterst,et al.  Quantitative telomerase enzyme activity determination using droplet digital PCR with single cell resolution , 2014, Nucleic acids research.

[15]  Xuezeng Zhao,et al.  Real-Time Monitoring of Insulin Using a Graphene Field-Effect Transistor Aptameric Nanosensor. , 2017, ACS applied materials & interfaces.

[16]  W. Lu,et al.  Improved synthesis of graphene oxide. , 2010, ACS nano.

[17]  Meng Liu,et al.  A graphene-based biosensing platform based on the release of DNA probes and rolling circle amplification. , 2014, ACS nano.

[18]  Jiani Xie,et al.  A universal strategy for aptamer-based nanopore sensing through host-guest interactions inside α-hemolysin. , 2015, Angewandte Chemie.

[19]  Jiye Shi,et al.  Graphene Nanoprobes for Real-Time Monitoring of Isothermal Nucleic Acid Amplification. , 2017, ACS Applied Materials and Interfaces.

[20]  Huang-Hao Yang,et al.  A graphene platform for sensing biomolecules. , 2009, Angewandte Chemie.

[21]  Andrea Galimberti,et al.  DNA Barcoding Meets Nanotechnology: Development of a Universal Colorimetric Test for Food Authentication. , 2017, Angewandte Chemie.

[22]  Xi Chen,et al.  Stabilization of ssRNA on graphene oxide surface: an effective way to design highly robust RNA probes. , 2013, Analytical chemistry.

[23]  Yuehe Lin,et al.  In situ simultaneous monitoring of ATP and GTP using a graphene oxide nanosheet–based sensing platform in living cells , 2014, Nature Protocols.

[24]  Dal‐Hee Min,et al.  Emerging Approaches for Graphene Oxide Biosensor. , 2017, Analytical chemistry.

[25]  Shouwu Guo,et al.  Adsorption of double-stranded DNA to graphene oxide preventing enzymatic digestion. , 2011, Nanoscale.

[26]  Meng Liu,et al.  Target-Induced and Equipment-Free DNA Amplification with a Simple Paper Device. , 2016, Angewandte Chemie.

[27]  Huang-Hao Yang,et al.  Using graphene to protect DNA from cleavage during cellular delivery. , 2010, Chemical communications.

[28]  Meng Liu,et al.  A DNAzyme Feedback Amplification Strategy for Biosensing. , 2017, Angewandte Chemie.

[29]  Kemin Wang,et al.  A versatile activatable fluorescence probing platform for cancer cells in vitro and in vivo based on self-assembled aptamer/carbon nanotube ensembles. , 2014, Analytical chemistry.

[30]  Wei Wen,et al.  Ultrasensitive Electrochemical Biosensor for HIV Gene Detection Based on Graphene Stabilized Gold Nanoclusters with Exonuclease Amplification. , 2015, ACS applied materials & interfaces.

[31]  Y. Duan,et al.  Amplified fluorescent aptasensor through catalytic recycling for highly sensitive detection of ochratoxin A. , 2015, Biosensors & bioelectronics.

[32]  Longhua Tang,et al.  Duplex DNA/Graphene Oxide Biointerface: From Fundamental Understanding to Specific Enzymatic Effects , 2012 .

[33]  D. L. Bain,et al.  Quantitative DNase footprint titration: a tool for analyzing the energetics of protein–DNA interactions , 2008, Nature Protocols.

[34]  Weihong Tan,et al.  Programmable and Multiparameter DNA-Based Logic Platform For Cancer Recognition and Targeted Therapy , 2014, Journal of the American Chemical Society.

[35]  Dong-Eun Kim,et al.  Mechanism of DNA adsorption and desorption on graphene oxide. , 2014, Langmuir : the ACS journal of surfaces and colloids.

[36]  A. Svatoš.  Single-cell metabolomics comes of age: new developments in mass spectrometry profiling and imaging. , 2011, Analytical chemistry.

[37]  Chad A Mirkin,et al.  Polyvalent DNA nanoparticle conjugates stabilize nucleic acids. , 2020, Nano letters.

[38]  Petr Skládal,et al.  Graphene oxide , 2022 .

[39]  D. Shank,et al.  Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Guo-Li Shen,et al.  Highly sensitive and selective bifunctional oligonucleotide probe for homogeneous parallel fluorescence detection of protein and nucleotide sequence. , 2011, Analytical chemistry.

[41]  Yi Lu,et al.  Label‐Free Colorimetric Detection of Lead Ions with a Nanomolar Detection Limit and Tunable Dynamic Range by using Gold Nanoparticles and DNAzyme , 2008 .

[42]  Jun Liu,et al.  Constraint of DNA on functionalized graphene improves its biostability and specificity. , 2010, Small.