Light triggered detection of aminophenyl phosphate with a quantum dot based enzyme electrode

An electrochemical sensor for p-aminophenyl phosphate (p APP) is reported. It is based on the electrochemical conversion of 4-aminophenol (4AP) at a quantum dot (QD) modified electrode under illumination. Without illumination no electron transfer and thus no oxidation of 4AP can occur. p APP as substrate is converted by the enzyme alkaline phosphatase (ALP) to generate 4AP as a product. The QDs are coupled via 1,4-benzenedithiol (BDT) linkage to the surface of a gold electrode and thus allow potential-controlled photocurrent generation. The photocurrent is modified by the enzyme reaction providing access to the substrate detection. In order to develop a photobioelectrochemical sensor the enzyme is immobilized on top of the photo-switchable layer of the QDs. Immobilization of ALP is required for the potential possibility of spatially resolved measurements. Geometries with immobilized ALP are compared versus having the ALP in solution. Data indicate that functional immobilization with layer-by-layer assembly is possible. Enzymatic activity of ALP and thus the photocurrent can be described by Michaelis- Menten kinetics. p APP is detected as proof of principle investigation within the range of 25 μM - 1 mM.

[1]  Wolfgang J. Parak,et al.  Investigation of the spatial resolution of the light-addressable potentiometric sensor , 2000 .

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

[3]  B. Limoges,et al.  Subfemtomolar Determination of Alkaline Phosphatase at a Disposable Screen-Printed Electrode Modified with a Perfluorosulfonated Ionomer Film , 1997 .

[4]  G. Urban,et al.  Determination of quantum confinement in CdSe nanocrystals by cyclic voltammetry , 2003 .

[5]  A. Pfreundt,et al.  Evaluation of quantum dots applied as switchable layer in a light-controlled electrochemical sensor , 2010, Analytical and bioanalytical chemistry.

[6]  A. Riedinger,et al.  In vitro and intracellular sensing by using the photoluminescence of quantum dots , 2010, Analytical and bioanalytical chemistry.

[7]  A. L. Latner,et al.  Isoelectric focusing of human liver alkaline phosphatase. , 1970, The Biochemical journal.

[8]  J. W. Parce,et al.  The cytosensor microphysiometer: biological applications of silicon technology. , 1992, Science.

[9]  Wolfgang J Parak,et al.  Labelling of cells with quantum dots , 2005, Nanotechnology.

[10]  X. Su,et al.  In vitro and in vivo imaging with quantum dots , 2010, Analytical and bioanalytical chemistry.

[11]  Louis E. Brus,et al.  The Quantum Mechanics of Larger Semiconductor Clusters ("Quantum Dots") , 1990 .

[12]  T. Nann,et al.  Heterogeneous charge transfer of colloidal nanocrystals in ionic liquids. , 2006, Chemphyschem : a European journal of chemical physics and physical chemistry.

[13]  A. Bodansky.  Phosphatase studies. 2. Determination of serum phosphatase. Factors influencing the accuracy of the determination. , 1933 .

[14]  Sander F. Wuister,et al.  Influence of thiol capping on the exciton luminescence and decay kinetics of CdTe and CdSe quantum dots , 2004 .

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

[16]  A. Seligman,et al.  The colorimetric determination of phosphatases in human serum. , 1951, The Journal of biological chemistry.

[17]  J. Emnéus,et al.  An enzyme flow immunoassay using alkaline phosphatase as the label and a tyrosinase biosensor as the label detector , 1998 .

[18]  J. Fischl,et al.  Microdetermination of phosphatases employing phenolphthalein diphosphate as substrate. , 1967, Clinical chemistry.

[19]  Christof M Niemeyer,et al.  Functional hybrid devices of proteins and inorganic nanoparticles. , 2003, Angewandte Chemie.

[20]  G. Siebert,et al.  Optischer Test zur Bestimmung der alkalischen Phosphatase im Serum , 1961, Klinische Wochenschrift.

[21]  D. Alpers,et al.  The two isozymes of rat intestinal alkaline phosphatase are products of two distinct genes. , 2000, Physiological genomics.

[22]  E. King,et al.  Estimation of Plasma Phosphatase by Determination of Hydrolysed Phenol with Amino-antipyrine , 1954, Journal of clinical pathology.

[23]  N. Tietz,et al.  An automated procedure for the determination of phosphorus and alkaline phosphatase (Bodansky) in serum. , 1964, Clinica chimica acta; international journal of clinical chemistry.

[24]  P. Talalay,et al.  SODIUM PHENOLPHTHALEIN PHOSPHATE AS A SUBSTRATE FOR PHOSPHATASE TESTS , 1945 .

[25]  Qiao Xu,et al.  Hydrogen peroxide biosensor based on direct electron transfer of horseradish peroxidase with vapor deposited quantum dots , 2009 .

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

[27]  W. Heineman,et al.  Competitive heterogeneous enzyme immunoassay for digoxin with electrochemical detection. , 1986, Analytical chemistry.

[28]  H. Neumann,et al.  An improved alkaline phosphatase determination with p-nitrophenyl phosphate. , 1967, Clinica chimica acta; international journal of clinical chemistry.

[29]  J. Frew,et al.  Measurement of alkaline phosphatase activity by electrochemical detection of phosphate esters: application to amperometric enzyme immunoassay , 1989 .

[30]  Waqas Khalid,et al.  Quantum-dot-modified electrode in combination with NADH-dependent dehydrogenase reactions for substrate analysis. , 2010, Langmuir : the ACS journal of surfaces and colloids.

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

[32]  Gero Decher,et al.  Multilayer Thin Films: Sequential Assembly of Nanocomposite Materials , 2003 .

[33]  U. Wollenberger,et al.  Biosensor for sensitive phosphate detection , 1992 .

[34]  W. Heineman,et al.  Comparison of methods for following alkaline phosphatase catalysis: spectrophotometric versus amperometric detection. , 1991, Analytical biochemistry.

[35]  Xiaoling Yang,et al.  Self-assembled CNTs/CdS/dehydrogenase hybrid-based amperometric biosensor triggered by photovoltaic effect. , 2008, Biosensors & bioelectronics.

[36]  E. King,et al.  A CONVENIENT METHOD FOR DETERMINING SERUM AND BILE PHOSPHATASE ACTIVITY. , 1934, Canadian Medical Association journal.

[37]  Jean-Louis Marty,et al.  Enzymatic recycling for signal amplification: Improving microcystin detection with biosensors , 2008 .

[38]  L. M. Jones,et al.  The estimation of serum inorganic phosphate and " acid " and " alkaline " phosphatase activity. , 1942 .

[39]  Mark A. Ratner,et al.  Electron transport in semiconducting carbon nanotubes with hetero-metallic contacts , 2005, cond-mat/0501538.

[40]  Shu-I Tu,et al.  1-Naphthyl phosphate as an enzymatic substrate for enzyme-linked immunomagnetic electrochemistry , 1999 .

[41]  W. Heineman,et al.  Competitive heterogeneous enzyme immunoassay for theophylline by flow-injection analysis with electrochemical detection of p-aminophenol. , 1990, Clinical chemistry.

[42]  Calum J. McNeil,et al.  Amperometric determination of alkaline phosphatase activity: application to enzyme immunoassay , 1987 .

[43]  G. Guilbault,et al.  Alkaline phosphatase as a label for immunoassay using amperometric detection with a variety of substrates and an optimal buffer system , 1999 .

[44]  T. Nann,et al.  Quantum confinement of the thermodynamic functions for the formation of electrons and holes in CdSe nanocrystals , 2006 .

[45]  W. Schuhmann,et al.  Redox-amplified biosensors based on selective modification of nanopore electrode structures with enzymes entrapped within electrodeposition paints , 2008 .

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

[47]  C. Niemeyer.  Functional Hybrid Devices of Proteins and Inorganic Nanoparticles , 2004 .

[48]  Gero Decher,et al.  Multilayer Thin Films , 2002 .

[49]  O. H. Lowry,et al.  A method for the rapid determination of alkaline phosphates with five cubic millimeters of serum. , 1946, The Journal of biological chemistry.

[50]  M. Dahan,et al.  Probing cellular events, one quantum dot at a time , 2010, Nature Methods.

[51]  R. Nitschke,et al.  Quantum dots versus organic dyes as fluorescent labels , 2008, Nature Methods.

[52]  S. Licht,et al.  A Light Addressable Photoelectrochemical Cyanide Sensor , 1996 .

[53]  L. Manna,et al.  Selective growth of PbSe on one or both tips of colloidal semiconductor nanorods. , 2005, Nano letters.

[54]  H. N. Fernley,et al.  Kinetic behaviour of calf-intestinal alkaline phosphatase with 4-methylumbelliferyl phosphate. , 1965, The Biochemical journal.

[55]  Wolfgang J. Parak,et al.  Lateral resolution of light-addressable potentiometric sensors: an experimental and theoretical investigation , 1997 .

[56]  William R. Heineman,et al.  p-aminophenyl phosphate: an improved substrate for electrochemical enzyme immnoassay , 1988 .

[57]  Jun Li,et al.  Direct electrochemistry of glucose oxidase and electrochemical biosensing of glucose on quantum dots/carbon nanotubes electrodes. , 2007, Biosensors & bioelectronics.

[58]  The integrated Michaelis-Menten equation. , 1962, Archives of biochemistry and biophysics.

[59]  F F Bier,et al.  Zeptomole-detecting biosensor for alkaline phosphatase in an electrochemical immunoassay for 2,4-dichlorophenoxyacetic acid. , 1996, Analytical chemistry.

[60]  Gero Decher,et al.  Fuzzy Nanoassemblies: Toward Layered Polymeric Multicomposites , 1997 .