Highly Sensitive Detection of Organophosphate Insecticides Using Biosensors Based on Genetically Engineered Acetylcholinesterase and Poly(3, 4-Ethylenedioxythiophene)

A poly(3,4-ethylenedioxythiophene) (PEDOT) conducting ink is presented as a new electroactive material to be incorporated in acetylcholinesterase-(AChE-) based screen printed biosensors, acting not only as a conducting template but also as an electrochemical mediator for thiocholine oxidation. Two different strategies have been studied for the chemical synthesis of PEDOT: (a) a classical oxidative polymerisation and (b) a more innovative enzymatic polymerisation, giving a water-soluble PEDOT. The use of this water-soluble conducting polymer as mediator in screen-printed biosensors enables its deposition by printing like the rest of the layers. Highly sensitive acetylcholinesterase-(AChE-) based screen-printed biosensors have been constructed using both classical and enzymatic PEDOT, in combination with genetically modified AChE. These electrodes allow the measurement of thiocholine oxidation at potentials of 100 mV versus Ag/AgCl reference electrode through the mediation of PEDOT. Inhibition of thiocholine production in presence of CPO allow for detection of this pesticide in concentrations as low as 1·10−10 M.

[1]  J. Anzai,et al.  Self-assembled biotinylated disulfide derivative monolayer on gold electrode for immobilizing enzymes. , 1997, Talanta.

[2]  J. Pomposo,et al.  First enzymatic synthesis of water-soluble conducting poly(3,4-ethylenedioxythiophene). , 2007, Biomacromolecules.

[3]  J. Pomposo,et al.  Ionic Liquid Immobilized Enzyme for Biocatalytic Synthesis of Conducting Polyaniline , 2006 .

[4]  L. Samuelson,et al.  The Role of Template in the Enzymatic Synthesis of Conducting Polyaniline , 1999 .

[5]  Y. Yoo,et al.  Synthesis of ortho-directed polyaniline using horseradish peroxidase , 2000 .

[6]  Robert Altenloh.  From a Novel , 1953 .

[7]  Till T Bachmann,et al.  Design of acetylcholinesterases for biosensor applications. , 2003, Biosensors & bioelectronics.

[8]  Jean-Louis Marty,et al.  Phosphotriesterase: a complementary tool for the selective detection of two organophosphate insecticides: chlorpyrifos and chlorfenvinfos. , 2009, Talanta.

[9]  P R Fielden,et al.  A microband lactate biosensor fabricated using a water-based screen-printed carbon ink. , 2009, Talanta.

[10]  Petr Skládal,et al.  Detection of organophosphate and carbamate pesticides using disposable biosensors based on chemically modified electrodes and immobilized cholinesterase , 1992 .

[11]  Damià Barceló,et al.  Evaluation of a highly sensitive amperometric biosensor with low cholinesterase charge immobilized on a chemically modified carbon paste electrode for trace determination of carbamates in fruit, vegetable and water samples , 1999 .

[12]  J. Hart,et al.  A novel, disposable, screen-printed amperometric biosensor for glucose in serum fabricated using a water-based carbon ink. , 2005, Biosensors & bioelectronics.

[13]  D. Barceló,et al.  Determination of carbamate residues in crop samples by cholinesterase-based biosensors and chromatographic techniques , 1998 .

[14]  Salvador Alegret,et al.  Determination of organophosphorus and carbamate pesticides using a biosensor based on a polishable, 7,7,8,8-tetracyanoquino-dimethane-modified, graphite—epoxy biocomposite , 1997 .

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

[16]  Petr Skládal,et al.  Detection of carbamate pesticides in vegetable samples using cholinesterase-based biosensors , 1997 .

[17]  S. Cosnier,et al.  Functionalized polypyrroles : a sophisticated glue for the immobilization and electrical wiring of enzymes , 1999 .

[18]  W. Marsden.  I and J , 2012 .

[19]  L. Samuelson,et al.  Mechanistic Study of the Peroxidase-Catalyzed Polymerization of Sulfonated Phenol† , 2001 .

[20]  J. Hart,et al.  Amperometric, screen-printed, glucose biosensor for analysis of human plasma samples using a biocomposite water-based carbon ink incorporating glucose oxidase. , 2005, Analytical biochemistry.

[21]  R. Pemberton,et al.  Fabrication of microband glucose biosensors using a screen-printing water-based carbon ink and their application in serum analysis. , 2009, Biosensors & bioelectronics.

[22]  J. Pomposo,et al.  Enzymatic Synthesis of Water-Soluble Conducting Poly(3,4- ethylenedioxythiophene): A Simple Enzyme Immobilization Strategy for Recycling and Reusing , 2009 .

[23]  L. Samuelson,et al.  Enzymatically Synthesized Conducting Polyaniline Nanocomposites: A Solid‐State NMR Study , 2003 .

[24]  L. Samuelson,et al.  Helical conformational specificity of enzymatically synthesized water-soluble conducting polyaniline nanocomposites. , 2003, Journal of the American Chemical Society.

[25]  Jean-Louis Marty,et al.  Screen-printed poly(3,4-ethylenedioxythiophene) (PEDOT): A new electrochemical mediator for acetylcholinesterase-based biosensors. , 2010, Talanta.

[26]  L. Samuelson,et al.  Manipulating DNA Conformation Using Intertwined Conducting Polymer Chains , 2001 .

[27]  Jean Louis Marty,et al.  Acetylcholine enzyme sensor for determining methamidophos insecticide: Evaluation of some genetically modified acetylcholinesterases from Drosophila melanogaster , 2001 .

[28]  H. Schreuder-Gibson,et al.  Conductive fibers from enzymatically synthesized polyaniline , 1999 .

[29]  J. Marty,et al.  Improved multianalyte detection of organophosphates and carbamates with disposable multielectrode biosensors using recombinant mutants of Drosophila acetylcholinesterase and artificial neural networks. , 2000, Biosensors & bioelectronics.

[30]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[31]  L. Samuelson,et al.  An Enzymatically Synthesized Conducting Molecular Complex of Polyaniline and Poly(vinylphosphonic acid) , 2000 .

[32]  Petr Skládal,et al.  Determination of organophosphate and carbamate pesticides using a cobalt phthalocyanine-modified carbon paste electrode and a cholinesterase enzyme membrane , 1991 .

[33]  D. Kaplan,et al.  Enzymatic catalysis in the synthesis of polyanilines and derivatives of polyanilines , 2006 .

[34]  A. Ciucu,et al.  Detection of pesticides using an amperometric biosensor based on ferophthalocyanine chemically modified carbon paste electrode and immobilized bienzymatic system. , 2003, Biosensors & bioelectronics.

[35]  Jean-Louis Marty,et al.  Adsorption: an easy and efficient immobilisation of acetylcholinesterase on screen-printed electrodes , 2003 .

[36]  M. Mascini,et al.  Sensitive detection of pesticides using amperometric sensors based on cobalt phthalocyanine-modified composite electrodes and immobilized cholinesterases. , 1992, Biosensors & bioelectronics.

[37]  J. Romero‐García,et al.  Template-free enzymatic synthesis of electrically conducting polyaniline using soybean peroxidase , 2005 .

[38]  Jean-Louis Marty,et al.  Immobilization of acetylcholinesterase on screen-printed electrodes: comparative study between three immobilization methods and applications to the detection of organophosphorus insecticides , 2002 .