Electrodeposited biotinylated polypyrrole as an immobilization method for impedimetric immunosensors

The potentialities of an electrodeposited biotinylated polypyrrole film as an immobilization matrix for the fabrication of impedimetric immunosensors are described. Biotinylated antibody (anti-human IgG), used as a model system, was attached to free biotin groups on the electrogenerated polypyrrole film using avidin as a coupling reagent. The resulting recognition interface consisted of a highly oriented monolayer immobilized onto the polymer surface. Cyclic voltammetry was used to characterize the polymer film. Additionally, scanning electron microscopy and atomic force microscopy were used to investigate the morphology of the immobilized material. This immobilization method allows the obtention of a highly reproducible and stable device. The resulting immunosensor has a linear dynamic range of 10-80 ng.ml/sup -1/ of antigen and a detection limit of 10 pg.ml/sup -1/. Furthermore, this immunosensor exhibited minor loss in response after two regeneration steps.

[1]  R. Redfield,et al.  Real-time biospecific interaction analysis of antibody reactivity to peptides from the envelope glycoprotein, gp160, of HIV-1. , 1992, Journal of immunological methods.

[2]  Dustin J Maly,et al.  Probing Low Affinity and Multivalent Interactions with Surface Plasmon Resonance: Ligands for Concanavalin A , 1998 .

[3]  W. Schuhmann,et al.  Synthesis of functionalized poly(dithienylpyrrole) derivatives and their application in amperometric biosensors , 1994 .

[4]  G G Guilbault,et al.  Recent developments in piezoelectric immunosensors. A review. , 1994, The Analyst.

[5]  S. Cosnier.  Biomolecule immobilization on electrode surfaces by entrapment or attachment to electrochemically polymerized films. A review. , 1999, Biosensors & bioelectronics.

[6]  Y. Hasebe,et al.  L‐Ascorbate sensor with polypyrrole‐coated carbon felt membrane electropolymerized in a cucumber juice solution , 1997 .

[7]  J. Therasse,et al.  Toxin detection using capacitance measurements on immunospecies grafted onto a semiconductor substrate , 1991 .

[8]  B. Scrosati,et al.  Properties of electrochemically synthesized polymer electrodes—IX. The effects of surfactants on polypyrrole films , 1992 .

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

[10]  L. Bachas,et al.  Biotin‐Modified Surfaces by Electrochemical Polymerizationof Biotinyl‐Tyramide , 1998 .

[11]  M. Aizawa.  Immunosensors. , 2019, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[12]  S. Cosnier,et al.  A biotinylated conducting polypyrrole for the spatially controlled construction of an amperometric biosensor. , 1999, Analytical chemistry.

[13]  A. Cass,et al.  Protein patterning with a photoactivatable derivative of biotin. , 1996, Bioconjugate chemistry.

[14]  R. Porter.  Investigation of electroplated conducting polymers as antibody receptors in immuno-sensors , 2000, Journal of immunoassay.

[15]  W. Schuhmann,et al.  A Low‐Volume Electrochemical Cell for the Deposition of Conducting Polymers and Entrapment of Enzymes , 1998 .

[16]  Anita Sargent,et al.  Monitoring antibody–antigen reactions at conducting polymer-based immunosensors using impedance spectroscopy , 1999 .

[17]  F. M. Starr.  Equivalent circuits , 1932, Electrical Engineering.

[18]  C. Maule,et al.  Detection and quantification of biomolecular interactions with optical biosensors , 1995 .

[19]  S. Cosnier,et al.  Poly(pyrrole–biotin): a new polymer for biomolecule grafting on electrode surfaces , 1999 .

[20]  F. Caruso,et al.  Surface chemical activation of quartz crystal microbalance gold electrodes — analysis by frequency changes, contact angle measurements and grazing angle FTIR , 1995 .

[21]  T F Christiansen,et al.  The slow penetration of enzymebased biosensors into clinical chemistry analysis , 1995, Acta anaesthesiologica Scandinavica. Supplementum.

[22]  M. Wilchek,et al.  Avidin-biotin technology ten years on: has it lived up to its expectations? , 1989, Trends in biochemical sciences.

[23]  Functionalized polypyrroles. New molecular materials for electrocatalysis and related applications , 1989 .

[24]  J. Diard,et al.  Impedance measurement errors due to non-linearities—I. Low frequency impedance measurements , 1994 .

[25]  Bo Johnsson,et al.  A novel hydrogel matrix on gold surfaces in surface plasmon resonance sensors for fast and efficient covalent immobilization of ligands , 1990 .

[26]  J. A. Logan,et al.  Conducting poly-N-alkylpyrrole polymer films , 1982 .

[27]  Ingemar Lundström,et al.  Real-time biospecific interaction analysis , 1994 .

[28]  R. Pei,et al.  Real-time immunoassay of antibody activity in serum by surface plasmon resonance biosensor. , 2000, Talanta.

[29]  Kazimierz Darowicki,et al.  The amplitude analysis of impedance spectra , 1995 .

[30]  H. Le,et al.  Human interleukin 4 receptor complex: neutralization effect of two monoclonal antibodies. , 1996, Biochemistry.

[31]  P. Ferloni,et al.  Impedance analysis of electronically conducting polymers , 1996 .

[32]  Boris Filanovsky,et al.  Photoswitchable Antigen−Antibody Interactions Studied by Impedance Spectroscopy , 1998 .

[33]  Nicole Jaffrezic-Renault,et al.  Gold electrode functionalized by electropolymerization of a cyano N-substituted pyrrole: application to an impedimetric immunosensor , 2001 .

[34]  B. Bauer.  Equivalent Circuit of , 1955 .