Label-free protein recognition using an aptamer-based impedance measurement assay

Abstract A novel aptamer-based biosensing assay for label-free protein detection and quantification by measuring the change in electrochemical impedance upon protein–aptamer complex formation was demonstrated. Our process starts with a self-assembly of the DNA aptamer on a microfabricated thin film gold electrode, followed by the recognition of the protein binding event via monitoring the interfacial electron transfer resistance with electrochemical impedance spectroscopy. The thrombin binding aptamer (TBA) has high binding affinity and specificity to its target protein and the impedance detection assay shows a reliable and sensitive quantification of thrombin with a detection range of four orders of magnitude and a detection limit of 0.1 nM.

[1]  I-Ming Hsing,et al.  Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection. , 2003, Lab on a chip.

[2]  M. Famulok,et al.  A Love-wave biosensor using nucleic acids as ligands , 2004 .

[3]  John Barford,et al.  Electrochemical detection of PCR amplicons using electroconductive polymer modified electrode and multiple nanoparticle labels , 2004 .

[4]  Friedrich C Simmel,et al.  A DNA-based machine that can cyclically bind and release thrombin. , 2004, Angewandte Chemie.

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

[6]  I-Ming Hsing,et al.  Enhanced electrochemical detection of DNA hybridization based on electrode-surface modification , 2003 .

[7]  Koji Sode,et al.  Novel electrochemical sensor system for protein using the aptamers in sandwich manner. , 2005, Biosensors & bioelectronics.

[8]  S. Swaminathan,et al.  A DNA aptamer which binds to and inhibits thrombin exhibits a new structural motif for DNA. , 1993, Biochemistry.

[9]  Makoto Shirai,et al.  Usage of a DNA Aptamer as a Ligand Targeting Microcystin , 2001 .

[10]  W. Tan,et al.  Molecular aptamer for real-time oncoprotein platelet-derived growth factor monitoring by fluorescence anisotropy. , 2001, Analytical chemistry.

[11]  J. Randles.  Kinetics of rapid electrode reactions , 1947 .

[12]  Yi Xiao,et al.  Aptamer-functionalized Au nanoparticles for the amplified optical detection of thrombin. , 2004, Journal of the American Chemical Society.

[13]  A. Ellington,et al.  Adapting selected nucleic acid ligands (aptamers) to biosensors. , 1998, Analytical chemistry.

[14]  L. Gold,et al.  The use of aptamers in large arrays for molecular diagnostics. , 1999, Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology.

[15]  Milan N Stojanovic,et al.  Fluorescent Sensors Based on Aptamer Self-Assembly. , 2000, Journal of the American Chemical Society.

[16]  S. Manalis,et al.  Micromechanical detection of proteins using aptamer-based receptor molecules. , 2004, Analytical chemistry.

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

[18]  S Tombelli,et al.  Development of biosensors with aptamers as bio-recognition element: the case of HIV-1 Tat protein. , 2004, Biosensors & bioelectronics.

[19]  Hans Wolf,et al.  An aptamer-based quartz crystal protein biosensor. , 2002, Analytical chemistry.

[20]  Chii Shang,et al.  Sequence-specific electrochemical recognition of multiple species using nanoparticle labels , 2004 .

[21]  A. Tulinsky,et al.  The structure of alpha-thrombin inhibited by a 15-mer single-stranded DNA aptamer. , 1994, The Journal of biological chemistry.

[22]  J. Feigon,et al.  Thrombin-binding DNA aptamer forms a unimolecular quadruplex structure in solution. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[23]  T. Yeates,et al.  Reconciliation of the X-ray and NMR structures of the thrombin-binding aptamer d(GGTTGGTGTGGTTGG). , 1996, Journal of molecular biology.

[24]  Nobuko Hamaguchi,et al.  Aptamer-based biosensor arrays for detection and quantification of biological macromolecules. , 2003, Analytical biochemistry.

[25]  E. Vermaas,et al.  Selection of single-stranded DNA molecules that bind and inhibit human thrombin , 1992, Nature.

[26]  Tibor Hianik,et al.  Detection of aptamer-protein interactions using QCM and electrochemical indicator methods. , 2005, Bioorganic & medicinal chemistry letters.

[27]  John T McDevitt,et al.  Aptamer-based sensor arrays for the detection and quantitation of proteins. , 2004, Analytical chemistry.

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

[29]  Weihong Tan,et al.  Molecular aptamer beacons for real-time protein recognition. , 2002, Biochemical and biophysical research communications.