An impedimetric immunosensor for the detection of autoantibodies directed against gliadins.

An immunosensor has been developed for the detection of autoantibodies directed against wheat gliadin, a protein fraction of cereal gluten which is involved in celiac disease. The immunosensor is based on the immobilization of gliadins onto gold electrodes covered with a polyelectrolyte layer of poly(4-styrenesulfonic acid sodium salt). The immobilization was monitored by quartz crystal microbalance (QCM) analysis. The antigen-antibody interaction signal was amplified by an incubation step with peroxidase-labeled immunoglobulins and subsequent peroxidase-catalyzed oxidation of 3-amino-9-ethylcarbazole (AEC). Changes in the insulating properties of the electrode layer were measured by electrochemical impedance spectroscopy (EIS) in the presence of ferri/ferro-cyanide. Impedance spectra could be fitted to a Randles equivalent circuit with high accuracy. Exposing the sensor electrodes to various antigliadin antibody concentrations resulted in proportional changes in the charge transfer resistance. A calibration graph for the detection of antigliadin antibodies was established for antibody concentrations between 10(-8) and 10(-6) M. Finally, the sensor was used for the determination of antigliadin autoantibodies of the IgG and IgA type in several human sera.

[1]  Yanjun Ding,et al.  Enzyme-catalyzed amplified immunoassay for the detection of Toxoplasma gondii-specific IgG using Faradaic impedance spectroscopy, CV and QCM , 2005, Analytical and bioanalytical chemistry.

[2]  W. Heineman,et al.  Small-volume voltammetric detection of 4-aminophenol with interdigitated array electrodes and its application to electrochemical enzyme immunoassay. , 1993, Analytical chemistry.

[3]  A. Görg,et al.  Electrophoretic characterization of wheat grain allergens from different cultivars involved in bakers' asthma , 1993, Electrophoresis.

[4]  F. Scheller,et al.  A superoxide sensor based on a multilayer cytochrome c electrode. , 2004, Analytical chemistry.

[5]  G. Sauerbrey.  Verwendung von Schwingquarzen zur Wägung dünner Schichten und zur Mikrowägung , 1959 .

[6]  R. Dietrich,et al.  Immunochemical detection of antibiotics and sulfonamides. , 1994, The Analyst.

[7]  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 .

[8]  F. Spener,et al.  An immunosensor based on disposable electrodes for rapid estimation of fatty acid-binding protein, an early marker of myocardial infarction. , 1997, Biosensors & bioelectronics.

[9]  Gang Wang,et al.  Application of impedance spectroscopy for monitoring colloid Au-enhanced antibody immobilization and antibody-antigen reactions. , 2004, Biosensors & bioelectronics.

[10]  Y. Shim,et al.  Development of an immunosensor for the detection of vitellogenin using impedance spectroscopy. , 2004, Biosensors & bioelectronics.

[11]  Y. Chai,et al.  A new label-free amperometric immunosenor for rubella vaccine , 2005, Analytical and bioanalytical chemistry.

[12]  S. Campuzano,et al.  An integrated electrochemical fructose biosensor based on tetrathiafulvalene-modified self-assembled monolayers on gold electrodes , 2003, Analytical and bioanalytical chemistry.

[13]  F. Caruso,et al.  Characterization of Polyelectrolyte-Protein Multilayer Films by Atomic Force Microscopy, Scanning Electron Microscopy, and Fourier Transform Infrared Reflection-Absorption Spectroscopy. , 1998 .

[14]  C. Fernández-Sánchez,et al.  AC voltammetric carbon paste-based enzyme immunosensors. , 2000, Biosensors & bioelectronics.

[15]  Yan Liu,et al.  New amperometric and potentiometric immunosensors based on gold nanoparticles/tris(2,2'-bipyridyl)cobalt(III) multilayer films for hepatitis B surface antigen determinations. , 2005, Biosensors & bioelectronics.

[16]  A. Offenhäusser,et al.  Functional tethered lipid bilayers. , 2000, Journal of biotechnology.

[17]  N. Jaffrezic‐Renault,et al.  Impedimetric immunosensor using avidin-biotin for antibody immobilization. , 2002, Bioelectrochemistry.

[18]  Petr Skládal,et al.  Advances in electrochemical immunosensors , 1997 .

[19]  O A Sadik,et al.  Applications of electrochemical immunosensors to environmental monitoring. , 1996, Biosensors & bioelectronics.

[20]  P. Schaaf,et al.  Protein Adsorption onto Auto-Assembled Polyelectrolyte Films , 2001 .

[21]  R. Hintsche,et al.  Rapid detection of viruses using electrical biochips and anti‐virion sera , 2005, Letters in applied microbiology.

[22]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

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

[24]  Angel Maquieira,et al.  On-line immunoanalysis for environmental pollutants: from batch assays to automated sensors , 1999 .

[25]  C. Egles,et al.  Multilayer Polyelectrolyte Films Functionalized by Insertion of Defensin: a New Approach to Protection of Implants from Bacterial Colonization , 2004, Antimicrobial Agents and Chemotherapy.

[26]  S. Kurosawa,et al.  Quartz crystal microbalance for the detection of microgram quantities of human serum albumin: relationship between the frequency change and the mass of protein adsorbed. , 1993, Analytical chemistry.

[27]  M. Thompson,et al.  Protein adsorption to organosiloxane surfaces studied by acoustic wave sensor. , 1998, The Analyst.

[28]  M. Thompson,et al.  Acoustic coupling of transverse waves as a mechanism for the label-free detection of protein-small molecule interactions. , 2002, The Analyst.

[29]  K R Rogers,et al.  Sol-gel-derived thick-film amperometric immunosensors. , 1998, Analytical chemistry.

[30]  O. Wolfbeis,et al.  Capacitive monitoring of protein immobilization and antigen-antibody reactions on monomolecular alkylthiol films on gold electrodes. , 1997, Biosensors & bioelectronics.

[31]  M vandeVen,et al.  Impedimetric immunosensors based on the conjugated polymer PPV. , 2005, Biosensors & bioelectronics.

[32]  G. Sauerbrey,et al.  Use of quartz vibration for weighing thin films on a microbalance , 1959 .

[33]  Vladimir Tvarozek,et al.  Optimization of capacitive affinity sensors: drift suppression and signal amplification , 1999 .

[34]  P. Schaaf,et al.  Human serum albumin self-assembly on weak polyelectrolyte multilayer films structurally modified by pH changes. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[35]  T. Kawai,et al.  SNPs Feasibility of Nonlabeled Oligonucletides on Using Electrochemical Sensing , 2004 .