A chronocoulometric aptasensor based on gold nanoparticles as a signal amplification strategy for detection of thrombin.

A sensitive chronocoulometric aptasensor for the detection of thrombin has been developed based on gold nanoparticle amplification. The functional gold nanoparticles, loaded with link DNA (LDNA) and report DNA (RDNA), were immobilized on an electrode by thrombin aptamers performing as a recognition element and capture probe. LDNA was complementary to the thrombin aptamers and RDNA was noncomplementary, but could combine with [Ru(NH₃)₆]³⁺ (RuHex) cations. Electrochemical signals obtained by RuHex that bound quantitatively to the negatively charged phosphate backbone of DNA via electrostatic interactions were measured by chronocoulometry. In the presence of thrombin, the combination of thrombin and thrombin aptamers and the release of the functional gold nanoparticles could induce a significant decrease in chronocoulometric signal. The incorporation of gold nanoparticles in the chronocoulometric aptasensor significantly enhanced the sensitivity. The performance of the aptasensor was further increased by the optimization of the surface density of aptamers. Under optimum conditions, the chronocoulometric aptasensor exhibited a wide linear response range of 0.1-18.5 nM with a detection limit of 30 pM. The results demonstrated that this nanoparticle-based amplification strategy offers a simple and effective approach to detect thrombin.

[1]  Y. Chai,et al.  Hemin/G-quadruplex simultaneously acts as NADH oxidase and HRP-mimicking DNAzyme for simple, sensitive pseudobienzyme electrochemical detection of thrombin. , 2012, Chemical communications.

[2]  Jing Wang,et al.  Gold nanoparticle enhanced electrochemiluminescence of CdS thin films for ultrasensitive thrombin detection. , 2011, Analytical chemistry.

[3]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[4]  Kyuwon Kim,et al.  Enzyme-amplified electrochemical detection of DNA using electrocatalysis of ferrocenyl-tethered dendrimer. , 2003, Analytical chemistry.

[5]  C. Fan,et al.  Electrochemical interrogation of DNA monolayers on gold surfaces. , 2005, Analytical chemistry.

[6]  Cristina Polonschii,et al.  A novel low-cost and easy to develop functionalization platform. Case study: aptamer-based detection of thrombin by surface plasmon resonance. , 2010, Talanta.

[7]  Joseph Wang,et al.  Electrochemical Aptasensors – Recent Achievements and Perspectives , 2009 .

[8]  Shuyan Niu,et al.  Fluorescence detection of thrombin using autocatalytic strand displacement cycle reaction and a dual-aptamer DNA sandwich assay. , 2012, Analytical biochemistry.

[9]  S. Yao,et al.  Ultrasensitive electrochemical aptasensor for thrombin based on the amplification of aptamer-AuNPs-HRP conjugates. , 2011, Biosensors & bioelectronics.

[10]  E. Wang,et al.  Solid-state label-free integrated aptasensor based on graphene-mesoporous silica-gold nanoparticle hybrids and silver microspheres. , 2011, Analytical Chemistry.

[11]  Feng Liu,et al.  Amplified QCM-D biosensor for protein based on aptamer-functionalized gold nanoparticles. , 2010, Biosensors & bioelectronics.

[12]  Itamar Willner,et al.  Nucleic acid-functionalized Pt nanoparticles: Catalytic labels for the amplified electrochemical detection of biomolecules. , 2006, Analytical chemistry.

[13]  Guo-Li Shen,et al.  Electrochemical aptameric recognition system for a sensitive protein assay based on specific target binding-induced rolling circle amplification. , 2010, Analytical chemistry.

[14]  Chad A Mirkin,et al.  Nanostructures in biodiagnostics. , 2005, Chemical reviews.

[15]  Qiang Zhao,et al.  Fluorescence anisotropy reduction of allosteric aptamer for sensitive and specific protein signaling. , 2012, Analytical chemistry.

[16]  Dayong Tian,et al.  Electrochemiluminescence biosensor for the assay of small molecule and protein based on bifunctional aptamer and chemiluminescent functionalized gold nanoparticles. , 2012, Analytica chimica acta.

[17]  Shouzhuo Yao,et al.  Sensitive bifunctional aptamer-based electrochemical biosensor for small molecules and protein. , 2009, Analytical chemistry.

[18]  Juewen Liu,et al.  Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes , 2006, Nature Protocols.

[19]  Feng Yan,et al.  Biomedical and clinical applications of immunoassays and immunosensors for tumor markers , 2007 .

[20]  D. Leech,et al.  Redox polymer and probe DNA tethered to gold electrodes for enzyme-amplified amperometric detection of DNA hybridization. , 2006, Analytical chemistry.

[21]  Wei-Wei Zhao,et al.  Double-probe signal enhancing strategy for toxin aptasensing based on rolling circle amplification. , 2012, Biosensors & bioelectronics.

[22]  Yan Wang,et al.  Aptamer-based highly sensitive electrochemical detection of thrombin via the amplification of graphene. , 2012, The Analyst.

[23]  Nicole Jaffrezic-Renault,et al.  A sensitive and selective thrombin impedimetric aptasensor based on tailored aptamers obtained by solid-phase synthesis , 2012 .

[24]  Yun Xiang,et al.  Quantum-dot/aptamer-based ultrasensitive multi-analyte electrochemical biosensor. , 2006, Journal of the American Chemical Society.

[25]  Y. Wan,et al.  An enzyme-based E-DNA sensor for sequence-specific detection of femtomolar DNA targets. , 2008, Journal of the American Chemical Society.

[26]  G. Shen,et al.  Aptamer-based rolling circle amplification: a platform for electrochemical detection of protein. , 2007, Analytical chemistry.

[27]  Chunhai Fan,et al.  Sequence-specific detection of femtomolar DNA via a chronocoulometric DNA sensor (CDS): effects of nanoparticle-mediated amplification and nanoscale control of DNA assembly at electrodes. , 2006, Journal of the American Chemical Society.

[28]  E. Wang,et al.  PVP-coated graphene oxide for selective determination of ochratoxin A via quenching fluorescence of free aptamer. , 2011, Biosensors & bioelectronics.

[29]  A. Steel,et al.  Electrochemical quantitation of DNA immobilized on gold. , 1998, Analytical chemistry.

[30]  Yue Zhao,et al.  Label-free electrochemiluminescent aptasensor with attomolar mass detection limits based on a Ru(phen)(3)(2+)-double-strand DNA composite film electrode. , 2009, Analytical chemistry.

[31]  Arben Merkoçi,et al.  Double-codified gold nanolabels for enhanced immunoanalysis. , 2007, Analytical chemistry.

[32]  Mi-Sook Won,et al.  Gold nanoparticles doped conducting polymer nanorod electrodes: ferrocene catalyzed aptamer-based thrombin immunosensor. , 2009, Analytical chemistry.

[33]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[34]  Defa Li,et al.  Ultrasensitive electrochemical detection of proteins by amplification of aptamer-nanoparticle bio bar codes. , 2007, Analytical chemistry.

[35]  Lianghai Hu,et al.  Aptamer in bioanalytical applications. , 2011, Analytical chemistry.