Quantum-dot-modified electrode in combination with NADH-dependent dehydrogenase reactions for substrate analysis.

A quantum dot-electrode system was developed which allows the sensitive detection of NADH (nicotinamide adenine dinucleotide). The colloidal semiconductive CdSe/ZnS nanocrystals (quantum dots) are attached to gold by chemisorption via a dithiol compound. The current signal can be triggered by illumination of the quantum-dot-modified electrode surface. Because of photoexcitation, electron-hole pairs are generated in the quantum dots, which can be detected as anodic or cathodic photocurrent. The immobilization of the nanocrystals is verified by amperometric photocurrent and quartz crystal microbalance (QCM) measurements. This study shows that CdSe/ZnS quantum dot-modified electrodes allow concentration dependent NADH detection in the range of 20 microM to 2 mM already at rather low potentials (around 0 V vs. Ag/AgCl, 1 M KCl). Therefore such electrodes can be used in combination with NADH-producing enzyme reactions for the light-triggered analysis of the respective substrates of the biocatalyst. It can be shown that glucose detection is feasible with such an electrode system and photocurrent measurements.

[1]  Tim Liedl,et al.  On the development of colloidal nanoparticles towards multifunctional structures and their possible use for biological applications. , 2004, Small.

[2]  Lo Gorton,et al.  Chemically modified electrodes for the electrocatalytic oxidation of nicotinamide coenzymes , 1986 .

[3]  I. Willner,et al.  Semiconductor quantum dots for bioanalysis. , 2008, Angewandte Chemie.

[4]  Xiaogang Peng,et al.  Experimental Determination of the Extinction Coefficient of CdTe, CdSe, and CdS Nanocrystals , 2003 .

[5]  Ioanis Katakis,et al.  Catalytic electrooxidation of NADH for dehydrogenase amperometric biosensors , 1997 .

[6]  Itamar Willner,et al.  Electroanalytical and Bioelectroanalytical Systems Based on Metal and Semiconductor Nanoparticles , 2004 .

[7]  Peter Reiss,et al.  Highly Luminescent CdSe/ZnSe Core/Shell Nanocrystals of Low Size Dispersion , 2002 .

[8]  L. Gorton,et al.  A comparative study of some 3,7-diaminophenoxazine derivatives and related compounds for electrocatalytic oxidation of NADH , 1990 .

[9]  Vincent M. Rotello,et al.  Applications of Nanoparticles in Biology , 2008 .

[10]  Itamar Willner,et al.  Photoelectrochemistry with Controlled DNA-Cross-Linked CdS Nanoparticle Arrays This research is supported by The U.S.-Israel Binational Science Foundation. The Max Planck Research Award for International Cooperation (I.W.) is gratefully acknowledged. , 2001, Angewandte Chemie.

[11]  T. Kuwana,et al.  Electrochemical stability of catechols with a pyrene side chain strongly adsorbed on graphite electrodes for catalytic oxidation of dihydronicotinamide adenine dinucleotide , 1983 .

[12]  B. Jena,et al.  Electrochemical biosensor based on integrated assembly of dehydrogenase enzymes and gold nanoparticles. , 2006, Analytical chemistry.

[13]  Lo Gorton,et al.  Carbon paste electrodes modified with enzymes, tissues, and cells , 1995 .

[14]  Itamar Willner,et al.  Nanoparticle–enzyme hybrid systems for nanobiotechnology , 2007, The FEBS journal.

[15]  L. Brus,et al.  Quantum crystallites and nonlinear optics , 1991 .

[16]  W. Blaedel,et al.  Study of the electrochemical oxidation of reduced nicotinamide adenine dinucleotide. , 1975, Analytical chemistry.

[17]  Liang Li,et al.  Core/Shell semiconductor nanocrystals. , 2009, Small.

[18]  Itamar Willner,et al.  Electrochemical control of the photocurrent direction in intercalated DNA/CdS nanoparticle systems. , 2005, Angewandte Chemie.

[19]  M Zanella,et al.  Photoelectrochemical signal chain based on quantum dots on gold--sensitive to superoxide radicals in solution. , 2008, Biosensors & bioelectronics.

[20]  A. Alivisatos,et al.  Molecular Weight, Osmotic Second Virial Coefficient, and Extinction Coefficient of Colloidal CdSe Nanocrystals , 2002 .

[21]  Joseph Wang Nanomaterial-based amplified transduction of biomolecular interactions. , 2005, Small.

[22]  L. Manna,et al.  Ligand exchange of CdSe nanocrystals probed by optical spectroscopy in the visible and mid-IR , 2008 .

[23]  Itamar Willner,et al.  Acetylcholine esterase-labeled CdS nanoparticles on electrodes: photoelectrochemical sensing of the enzyme inhibitors. , 2003, Journal of the American Chemical Society.

[24]  A Paul Alivisatos,et al.  Room-temperature single-nucleotide polymorphism and multiallele DNA detection using fluorescent nanocrystals and microarrays. , 2003, Analytical chemistry.

[25]  Wolfgang J Parak,et al.  Quantum dots on gold: electrodes for photoswitchable cytochrome C electrochemistry. , 2006, Small.

[26]  V. Flexer,et al.  Wired-enzyme core-shell Au nanoparticle biosensor. , 2008, Journal of the American Chemical Society.

[27]  Xiliang Luo,et al.  Nanocomposite and nanoporous polyaniline conducting polymers exhibit enhanced catalysis of nitrite reduction. , 2007, Chemistry.

[28]  Itamar Willner,et al.  Controlling the direction of photocurrents by means of CdS nanoparticles and cytochrome c-mediated biocatalytic cascades. , 2006, Chemical communications.

[29]  Igor L. Medintz,et al.  Multiplexed toxin analysis using four colors of quantum dot fluororeagents. , 2004, Analytical chemistry.

[30]  Itamar Willner,et al.  Halbleiterquantenpunkte für die Bioanalyse , 2008 .

[31]  Haoqing Hou,et al.  Electrospun Palladium Nanoparticle‐Loaded Carbon Nanofibers and Their Electrocatalytic Activities towards Hydrogen Peroxide and NADH , 2008 .

[32]  Helmuth Möhwald,et al.  Self-assembly of electro-active protein architectures on electrodes for the construction of biomimetic signal chains. , 2009, Chemical communications.

[33]  S. Fukuzumi,et al.  Mechanism of hydride transfer from an NADH model compound to p-benzoquinone derivatives , 1984 .

[34]  K. Fujioka,et al.  Simultaneous Multicolor Detection System of the Single‐Molecular Microbial Antigen with Total Internal Reflection Fluorescence Microscopy , 2005, Microbiology and immunology.

[35]  A. Alivisatos Semiconductor Clusters, Nanocrystals, and Quantum Dots , 1996, Science.

[36]  M. Bawendi,et al.  (CdSe)ZnS Core-Shell Quantum Dots - Synthesis and Characterization of a Size Series of Highly Luminescent Nanocrystallites , 1997 .

[37]  Igor L. Medintz,et al.  Biosensing with Luminescent Semiconductor Quantum Dots , 2006, Sensors (Basel, Switzerland).

[38]  Warren C. W. Chan,et al.  Quantum Dots in Biological and Biomedical Research: Recent Progress and Present Challenges , 2006 .

[39]  Wolfgang Schuhmann,et al.  Electrocatalytic oxidation of reduced nicotinamide coenzymes at gold and platinum electrode surfaces modified with a monolayer of pyrroloquinoline quinone. Effect of Ca2+ cations , 1994 .

[40]  N. Mano,et al.  Mediator-modified electrodes for catalytic NADH oxidation: high rate constants at interesting overpotentials. , 2002, Bioelectrochemistry.

[41]  B. Jena,et al.  Efficient electrocatalytic oxidation of NADH at gold nanoparticles self-assembled on three-dimensional sol-gel network. , 2005, Chemical communications.

[42]  Rosario Pereiro,et al.  Bioanalytics and biolabeling with semiconductor nanoparticles (quantum dots) , 2007 .

[43]  Yinghong Xiao,et al.  Nanocomposites: From Fabrications to Electrochemical Bioapplications , 2008 .

[44]  Miltiades I. Karayannis,et al.  Enzyme Based Amperometric Biosensors for Food Analysis , 2002 .

[45]  Itamar Willner,et al.  Self-assembly of semiconductor quantum-dots on electrodes for photoelectrochemical biosensing , 2007, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[46]  S. Maenosono,et al.  Photoinduced fluorescence intensity oscillation in a reaction-diffusion cell containing a colloidal quantum dot dispersion. , 2006, The Journal of chemical physics.

[47]  S. Nie,et al.  Luminescent quantum dots for multiplexed biological detection and imaging. , 2002, Current opinion in biotechnology.

[48]  Da Chen,et al.  Interfacial Bioelectrochemistry: Fabrication, Properties and Applications of Functional Nanostructured Biointerfaces , 2007 .

[49]  Joseph Wang,et al.  Electrochemistry of nicotinamide adenine dinucleotide (reduced) at heated platinum electrodes , 2005 .

[50]  S. Pathak,et al.  Hydroxylated quantum dots as luminescent probes for in situ hybridization. , 2001, Journal of the American Chemical Society.

[51]  I. Willner,et al.  NAD(+)/NADH-sensitive quantum dots: applications to probe NAD(+)-dependent enzymes and to sense the RDX explosive. , 2009, Nano letters.