Frequency-dependent electrical detection of protein binding events.

Frequency-dependent electrochemical impedance spectroscopy has been used to characterize the changes in electrical response that accompany specific binding of a protein to its substrate, using the biotin-avidin system as a model. Our results show that avidin, at concentrations in the nanomolar range, can be detected electrically in a completely label-free manner under conditions of zero average current flow and without the use of any auxiliary redox agents. Impedance measurements performed on biotin-modified surfaces of gold, glassy carbon, and silicon were obtained over a wide frequency range, from 5 mHz to 1 MHz. On each biotin-modified surface, binding of avidin is most easily detected at low frequencies, <1 Hz. Electrical circuit modeling of the interface was used to relate the frequency-dependent electrical response to the physical structure of the interface before and after avidin binding. Electrical measurements were correlated with measurements of protein binding using fluorescently labeled avidin.

[1]  R. Bruce Lennox,et al.  Stability of ω-Functionalized Self-Assembled Monolayers as a Function of Applied Potential , 2000 .

[2]  A. Blanchard,et al.  High-density oligonucleotide arrays , 1996 .

[3]  J. Xu,et al.  Anthraquinonedisulfonate electrochemistry:  a comparison of glassy carbon, hydrogenated glassy carbon, highly oriented pyrolytic graphite, and diamond electrodes. , 1998, Analytical chemistry.

[4]  H. Takenouti,et al.  Long-time and short-time investigation of the electrode interface through electrochemical impedance measurements. Application to adsorption of human serum albumin onto glassy carbon rotating disc electrode , 1997 .

[5]  A. Bard,et al.  Immobilization and Hybridization of DNA on an Aluminum(III) Alkanebisphosphonate Thin Film with Electrogenerated Chemiluminescent Detection , 1995 .

[6]  Itamar Willner,et al.  The Use of Impedance Spectroscopy for the Characterization of Protein-Modified ISFET Devices: Application of the Method for the Analysis of Biorecognition Processes , 2001 .

[7]  J. N. Russell,et al.  Photochemical Functionalization of Diamond Films , 2002 .

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

[9]  E. Southern,et al.  A novel method for the analysis of multiple sequence variants by hybridisation to oligonucleotides. , 1993, Nucleic acids research.

[10]  Elizabeth M. Boon,et al.  Mutation detection by electrocatalysis at DNA-modified electrodes , 2000, Nature Biotechnology.

[11]  M. Hill,et al.  Long-Range Electron Transfer through DNA Films. , 1999, Angewandte Chemie.

[12]  M F Lawrence,et al.  Immobilization of homooligonucleotide probe layers onto Si/SiO(2) substrates: characterization by electrochemical impedance measurements and radiolabelling. , 2002, Biosensors & bioelectronics.

[13]  Nam Quoc Ngo,et al.  The Efficiency of Light-Directed Synthesis of DNA Arrays on Glass Substrates , 1997 .

[14]  E. Souteyrand,et al.  DIRECT DETECTION OF THE HYBRIDIZATION OF SYNTHETIC HOMO-OLIGOMER DNA SEQUENCES BY FIELD EFFECT , 1997 .

[15]  Lloyd M. Smith,et al.  Synthesis and Characterization of DNA-Modified Silicon (111) Surfaces , 2000 .

[16]  W. Göpel,et al.  Impedance spectroscopy and scanning tunneling microscopy of polished and electrochemically pretreated glassy carbon , 1994 .

[17]  T. D. Harris,et al.  Measuring the structure of etched silicon surfaces with Raman spectroscopy , 1994 .

[18]  Anthony G. Frutos,et al.  Surface plasmon resonance imaging measurements of DNA hybridization adsorption and streptavidin/DNA multilayer formation at chemically modified gold surfaces , 1997 .

[19]  A. Steel,et al.  Electrostatic interactions of redox cations with surface-immobilized and solution DNA. , 1999, Bioconjugate chemistry.

[20]  Juan Bisquert,et al.  Doubling Exponent Models for the Analysis of Porous Film Electrodes by Impedance. Relaxation of TiO2 Nanoporous in Aqueous Solution , 2000 .

[21]  T. M. Herne,et al.  Observation of Hybridization and Dehybridization of Thiol-Tethered DNA Using Two-Color Surface Plasmon Resonance Spectroscopy , 1997 .

[22]  S. Dong,et al.  A facile approach to immobilize protein for biosensor: self-assembled supported bilayer lipid membranes on glassy carbon electrode. , 2001, Biosensors & bioelectronics.

[23]  M. Schöning,et al.  Recent advances in biologically sensitive field-effect transistors (BioFETs). , 2002, The Analyst.

[24]  Z. Cheng,et al.  Ion channel behavior of supported bilayer lipid membranes on a glassy carbon electrode. , 2000, Analytical chemistry.

[25]  C. Padeste,et al.  Ferrocene-avidin conjugates for bioelectrochemical applications. , 2000, Biosensors & bioelectronics.

[26]  P Bergveld,et al.  Development of an ion-sensitive solid-state device for neurophysiological measurements. , 1970, IEEE transactions on bio-medical engineering.

[27]  Jacqueline K. Barton,et al.  Oxidative DNA damage through long-range electron transfer , 1996, Nature.

[28]  B. Boukamp,et al.  Effect of Hexacyanoferrate(II/III) on Self-Assembled Monolayers of Thioctic Acid and 11-Mercaptoundecanoic Acid on Gold , 2002 .

[29]  S. P. Fodor,et al.  Multiplexed biochemical assays with biological chips , 1993, Nature.

[30]  Min Guo,et al.  Characterization of defects in the formation process of self-assembled thiol monolayers by electrochemical impedance spectroscopy , 2001 .

[31]  W. Everett,et al.  Factors that influence the stability of self-assembled organothiols on gold under electrochemical conditions , 1995 .