Label-free impedimetric immunosensor for sensitive detection of ochratoxin A.

A novel label-free electrochemical impedimetric immunosensor for sensitive detection of ochratoxin A (OTA) was reported. A two-step reaction protocol was elaborated to modify the gold electrode. The electrode was first derivatized by electrochemical reduction of in situ generated 4-carboxyphenyl diazonium salt (4-CPDS) in acidic aqueous solution yielded stable 4-carboxyphenyl (4-CP) monolayer. The ochratoxin A antibody was then immobilized making use of the carbodiimide chemistry. The steps of the immunosensor elaboration and the immunochemical reaction between ochratoxin A and the surface-bound antibody were interrogated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The impedance change, due to the specific immuno-interaction at the immunosensor surface was utilized to detect ochratoxin A. The increase in electron-transfer resistance (DeltaR(et)) values was linearly proportional to the concentration of OTA in the range of 1-20ngmL(-1), with a detection limit of 0.5ngmL(-1).

[1]  J. Pinson,et al.  Attachment of organic layers to conductive or semiconductive surfaces by reduction of diazonium salts. , 2005, Chemical Society reviews.

[2]  Serge Cosnier,et al.  Affinity Biosensors Based on Electropolymerized Films , 2005 .

[3]  R. Harvey,et al.  Immunotoxicity of ochratoxin A to growing gilts. , 1992, American journal of veterinary research.

[4]  A. Merkoçi,et al.  Electrochemical genosensor design: immobilisation of oligonucleotides onto transducer surfaces and detection methods. , 2000, Biosensors & bioelectronics.

[5]  G. Whitesides,et al.  Patterning ligands on reactive SAMs by microcontact printing , 1999 .

[6]  J. Pinson,et al.  Immobilization of glucose oxidase on a carbon surface derivatized by electrochemical reduction of diazonium salts , 1992 .

[7]  Z. Li,et al.  Surface modification and functionalization through the self-assembled monolayer and graft polymerization. , 2005, Advances in colloid and interface science.

[8]  R. Leblanc,et al.  Electron-transfer properties of cytochrome c Langmuir-Blodgett films and interactions of cytochrome c with lipids , 1998 .

[9]  Nicole Jaffrezic-Renault,et al.  A disposable immunomagnetic electrochemical sensor based on functionalised magnetic beads on gold surface for the detection of atrazine , 2006 .

[10]  N. Pourmand,et al.  Label-Free Impedance Biosensors: Opportunities and Challenges. , 2007, Electroanalysis.

[11]  Daniel G. Pinacho,et al.  Impedimetric immunosensor for the specific label free detection of ciprofloxacin antibiotic. , 2007, Biosensors & bioelectronics.

[12]  R. Nezlin.  The Immunoglobulins: Structure and Function , 1998 .

[13]  Elena Lizarraga,et al.  Comparison between capillary electrophoresis and HPLC-FL for ochratoxin A quantification in wine , 2006 .

[14]  M. Feng,et al.  Direct electrochemistry and Raman spectroscopy of sol-gel-encapsulated myoglobin. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[15]  M. Váradi,et al.  Development of immunosensor based on OWLS technique for determining Aflatoxin B1 and Ochratoxin A. , 2007, Biosensors & bioelectronics.

[16]  E. Barsoukov,et al.  Impedance spectroscopy : theory, experiment, and applications , 2005 .

[17]  T. M. Nahir,et al.  Impedance Spectroscopy: Theory, Experiment, and Applications, 2nd ed Edited by Evgenij Barsoukov (Texas Instruments Inc.) and J. Ross Macdonald (University of North Carolina, Chapel Hill). John Wiley & Sons, Inc.: Hoboken, NJ. 2005. xvii + 596 pp. $125.00. ISBN 0471-64749-7. , 2005 .

[18]  Shawn M. Dirk,et al.  Electroaddressable Selective Functionalization of Electrode Arrays: Catalytic NADH Detection Using Aryl Diazonium Modified Gold Electrodes , 2007 .

[19]  H Valenta,et al.  Chromatographic methods for the determination of ochratoxin A in animal and human tissues and fluids. , 1998, Journal of chromatography. A.

[20]  I. Willner,et al.  Probing Biomolecular Interactions at Conductive and Semiconductive Surfaces by Impedance Spectroscopy: Routes to Impedimetric Immunosensors, DNA‐Sensors, and Enzyme Biosensors , 2003 .

[21]  T. Lea,et al.  Effects of ochratoxin A upon early and late events in human T-cell proliferation. , 1995, Toxicology.

[22]  Yanli Zhou,et al.  Development of an amperometric biosensor based on covalent immobilization of tyrosinase on a boron-doped diamond electrode , 2006 .

[23]  D. Bélanger,et al.  Characterization of the deposition of organic molecules at the surface of gold by the electrochemical reduction of aryldiazonium cations. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[24]  G. Dirheimer,et al.  In vitro DNA and dGMP adducts formation caused by ochratoxin A. , 2000, Chemico-biological interactions.

[25]  R. D. Whitley,et al.  Ion-exchange equilibria of lysozyme, myoglobin and bovine serum albumin. Effective valence and exchanger capacity. , 1989, Journal of chromatography.

[26]  J. Pinson,et al.  Organic Layers Bonded to Industrial, Coinage, and Noble Metals through Electrochemical Reduction of Aryldiazonium Salts , 2003 .

[27]  Ian Ivar Suni,et al.  Electrochemical Impedance Biosensor for Glucose Detection Utilizing a Periplasmic E. coli Receptor Protein , 2005 .

[28]  M. Prodromidis,et al.  Development of a faradic impedimetric immunosensor for the detection of Salmonella typhimurium in milk. , 2008, Analytical chemistry.

[29]  C. Schlatter,et al.  Carcinogenicity and kinetic aspects of ochratoxin A. , 1996, Food additives and contaminants.

[30]  Yukio Saito,et al.  Protein-binding of ochratoxin A and its extractability from proteinous food , 1985 .

[31]  A. Visconti,et al.  Determination of ochratoxin A in domestic and imported beers in italy by immunoaffinity clean-up and liquid chromatography. , 2000, Journal of chromatography. A.

[32]  James F. Rusling,et al.  Direct Electrochemistry of Myoglobin and Cytochrome P450cam in Alternate Layer-by-Layer Films with DNA and Other Polyions , 1998 .

[33]  G. Palleschi,et al.  Electrochemical oxidation of ochratoxin A at a glassy carbon electrode and in situ evaluation of the interaction with deoxyribonucleic acid using an electrochemical deoxyribonucleic acid-biosensor. , 2007, Analytica chimica acta.

[34]  F. Minisci.  Diazo Chemistry I. Aromatic and Heteroaromatic Compounds , 1995 .

[35]  Frank Davis,et al.  Label-less immunosensor assay for myelin basic protein based upon an ac impedance protocol. , 2008, Analytical chemistry.

[36]  D. J. Schuller,et al.  Crystal structure of horseradish peroxidase C at 2.15 Å resolution , 1997, Nature Structural Biology.

[37]  Miriam M. Ngundi,et al.  Rapid detection of foodborne contaminants using an Array Biosensor , 2006 .

[38]  J. Gooding,et al.  The modification of glassy carbon and gold electrodes with aryl diazonium salt: The impact of the electrode materials on the rate of heterogeneous electron transfer , 2005 .

[39]  J. Pitt.  Toxigenic fungi and mycotoxins. , 2000, British medical bulletin.

[40]  F. Lisdat,et al.  Screen-printed electrodes as impedimetric immunosensors for the detection of anti-transglutaminase antibodies in human sera. , 2007, Analytica chimica acta.

[41]  P. Skládal,et al.  The immunosensors for measurement of 2,4-dichlorophenoxyacetic acid based on electrochemical impedance spectroscopy. , 2004, Bioelectrochemistry.

[42]  H. Köhler,et al.  Studies of the influence of ochratoxin A on immune and defence reactions in weaners , 1999, Mycoses.

[43]  Muhammad J A Shiddiky,et al.  An impedimetric immunosensor for the label-free detection of bisphenol A. , 2007, Biosensors & bioelectronics.

[44]  Giuseppe Palleschi,et al.  Monoclonal antibody based electrochemical immunosensor for the determination of ochratoxin A in wheat. , 2006, Talanta.

[45]  Heidi R. C. Dietrich,et al.  Biosensors and multiple mycotoxin analysis , 2003 .

[46]  Raju Khan,et al.  Nanocrystalline bioactive TiO2–chitosan impedimetric immunosensor for ochratoxin-A , 2008 .

[47]  Jean-Louis Marty,et al.  Novel highly-performing immunosensor-based strategy for ochratoxin A detection in wine samples. , 2008, Biosensors & bioelectronics.

[48]  Itamar Willner,et al.  Enzyme-Linked Amplified Electrochemical Sensing of Oligonucleotide−DNA Interactions by Means of the Precipitation of an Insoluble Product and Using Impedance Spectroscopy , 1999 .

[49]  Richard O'Kennedy,et al.  The development of a 'labeless' immunosensor for the detection of Listeria monocytogenes cell surface protein, Internalin B. , 2008, Biosensors & bioelectronics.

[50]  D. Bélanger,et al.  Electrochemical derivatization of carbon surface by reduction of in situ generated diazonium cations. , 2005, The journal of physical chemistry. B.

[51]  B. Limoges,et al.  BIOTINYLATION OF SCREEN-PRINTED CARBON ELECTRODES THROUGH THE ELECTROCHEMICAL REDUCTION OF THE DIAZONIUM SALT OF P-AMINOBENZOYL BIOCYTIN , 1999 .

[52]  J. Gooding,et al.  Diazonium salts: Stable monolayers on gold electrodes for sensing applications , 2007 .

[53]  Jian Wang,et al.  Covalent immobilization of glucose oxidase on conducting ultrananocrystalline diamond thin films , 2006 .

[54]  J. Pinson,et al.  Covalent Modification of Carbon Surfaces by Aryl Radicals Generated from the Electrochemical Reduction of Diazonium Salts , 1997 .

[55]  K. Eichmann,et al.  INDUCTION OF RABBIT ANTIBODY WITH MOLECULAR UNIFORMITY AFTER IMMUNIZATION WITH GROUP C STREPTOCOCCI , 1970, The Journal of experimental medicine.

[56]  A. Downard,et al.  Grafting Aryl Diazonium Cations to Polycrystalline Gold: Insights into Film Structure Using Gold Oxide Reduction, Redox Probe Electrochemistry, and Contact Angle Behavior , 2007 .

[57]  S. B. Hall,et al.  Electrochemical Reduction of a Conjugated Cinnamic Acid Diazonium Salt as an Immobilization Matrix for Glucose Biosensor , 2003 .