TY - JOUR A1 - Kapp, A. A1 - Beissenhirtz, Moritz Karl A1 - Geyer, F. A1 - Scheller, F. A1 - Viezzoli, Maria Silvia A1 - Lisdat, Fred T1 - Electrochemical and sensorial behavior of SOD mutants immobilized on gold electrodes in aqueous/organic solvent mixtures JF - Electroanalysis : an international journal devoted to fundamental and practical aspects of electroanalysis N2 - A cysteine mutant of a monomeric human Cu, Zn-SOD (Glycine 61, Serine 142) has been immobilized directly on gold electrodes using the thiol groups introduced. The electrochemical behavior of the surface confined protein was studied in mixtures of aqueous buffer and DMSO up to an organic solvent content of 60%. The formal potential was found to be rather independent of the DMSO content. However, half peak width increased and the redoxactive amount clearly decreased with raising DMSO content. In addition, the kinetics of the heterogeneous electron transfer became slower; but still a quasireversible electrochemical conversion of the mutant SOD was feasible. Thus, the electrodes were applied for sensorial superoxide detection. At a potential of +220 mV vs. Ag/AgCl advantage was taken of the partial oxidation reaction of the enzyme. A defined superoxide signal was obtained in solutions up to 40% DMSO. The sensitivity of the mutant electrodes decreased linearly with the organic solvent content in solution but was still higher compared to conventional cyt.c based sensors. At DMSO concentrations higher than 40% no sensor response was detected. KW - SOD KW - mutants KW - gold electrodes KW - DMSO KW - electrochemistry Y1 - 2006 U6 - https://doi.org/10.1002/elan.200603620 SN - 1040-0397 VL - 18 SP - 1909 EP - 1915 PB - Wiley CY - Weinheim ER - TY - GEN A1 - Spricigo, Roberto A1 - Dronov, Roman A1 - Lisdat, Fred A1 - Leimkühler, Silke A1 - Scheller, Frieder W. A1 - Wollenberger, Ursula T1 - Electrocatalytic sulfite biosensor with human sulfite oxidase co-immobilized with cytochrome c in a polyelectrolyte-containing multilayer T2 - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - An efficient electrocatalytic biosensor for sulfite detection was developed by co-immobilizing sulfite oxidase and cytochrome c with polyaniline sulfonic acid in a layer-by-layer assembly. QCM, UV-Vis spectroscopy and cyclic voltammetry revealed increasing loading of electrochemically active protein with the formation of multilayers. The sensor operates reagentless at low working potential. A catalytic oxidation current was detected in the presence of sulfite at the modified gold electrode, polarized at +0.1 V ( vs. Ag/AgCl 1 M KCl). The stability of the biosensor performance was characterized and optimized. A 17-bilayer electrode has a linear range between 1 and 60 mu M sulfite with a sensitivity of 2.19 mA M-1 sulfite and a response time of 2 min. The electrode retained a stable response for 3 days with a serial reproducibility of 3.8% and lost 20% of sensitivity after 5 days of operation. It is possible to store the sensor in a dry state for more than 2 months. The multilayer electrode was used for determination of sulfite in unspiked and spiked samples of red and white wine. The recovery and the specificity of the signals were evaluated for each sample. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 945 KW - bioelectrocatalysis KW - sulfite KW - sulfite oxidase KW - cytochrome c KW - multilayer Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-431176 SN - 1866-8372 IS - 945 SP - 225 EP - 233 ER - TY - JOUR A1 - Kapp, Andreas A1 - Beissenhirtz, Moritz Karl A1 - Geyer, F. A1 - Scheller, Frieder W. A1 - Viezzoli, Maria Silvia A1 - Lisdat, Fred T1 - Electrochemical and sensorial behaviour of SOD mutants immobilized on gold electrodes in aqueous / organic solvent mixtures Y1 - 2006 UR - http://www3.interscience.wiley.com/cgi-bin/jhome/26571/ U6 - https://doi.org/10.1002/elan.200603620 SN - 1040-0397 ER - TY - JOUR A1 - Beissenhirtz, Moritz Karl A1 - Scheller, Frieder W. A1 - Viezzoli, Maria Silvia A1 - Lisdat, Fred T1 - Engineered superoxide dismutase monomers for superoxide biosensor applications N2 - Because of its high reaction rate and specificity, the enzyme superoxide dismutase (SOD) offers great potential for the sensitive quantification of superoxide radicals in electrochemical biosensors. In this work, monomeric mutants of human Cu,Zn-SOD were engineered to contain one or two additional cysteine residues, which could be used to bind the protein to gold surfaces, thus making the use of promotor molecules unnecessary. Six mutants were successfully designed, expressed, and purified. All mutants bound directly to unmodified gold surfaces via the sulfur of the cysteine residues and showed a quasireversible, direct electron transfer to the electrode. Thermodynamic and kinetic parameters of the electron transfer were characterized and showed only slight variations between the individual mutants. For one of the mutants, the interaction with the superoxide radical was studied in more detail. For both partial reactions of the dismutation, an interaction between protein and radical could be shown. In an amperometric biosensorial approach, the SOD-mutant electrode was successfully applied for the detection of superoxide radicals. In the oxidation region, the electrode surpassed the sensitivity of the commonly used cytochrome c electrodes by similar to 1 order of magnitude while not being limited by interferences, but the electrode did not fully reach the sensitivity of dimeric Cu,Zn-SOD immobilized on MPA-modified gold Y1 - 2006 UR - http://pubs.acs.org/journal/ancham U6 - https://doi.org/10.1021/Ac051465g SN - 0003-2700 ER - TY - JOUR A1 - Beissenhirtz, Moritz Karl A1 - Scheller, Frieder W. A1 - Stöcklein, Walter F. M. A1 - Kurth, D. A1 - Möhwald, Helmuth A1 - Lisdat, Fred T1 - Electroactive cytochrome c multilayers within a polyelectrolyte assembly Y1 - 2004 ER - TY - JOUR A1 - Kröning, Steffen A1 - Scheller, Frieder W. A1 - Wollenberger, Ursula A1 - Lisdat, Fred T1 - Myoglobin-Clay Electrode for Nitric Oxide (NO) Detection in Solution Y1 - 2004 ER - TY - JOUR A1 - Scheller, Frieder W. A1 - Lisdat, Fred A1 - Wollenberger, Ursula T1 - Application of electrically contacted enzymes for biosensors Y1 - 2005 SN - 3-527- 30690-0 ER - TY - JOUR A1 - Krylov, Andrey V. A1 - Beissenhirtz, Moritz Karl A1 - Adamzig, Holger A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - Thick-film electrodes for measurement of superoxide and hydrogen peroxide based on direct protein-electrode contacts N2 - Cytochrome c was immobilized on screen-printed thick-film gold electrodes by a self-assembly approach using mixed monolayers of mercaptoundecanoic acid and mercaptoundecanol. Cyclic voltammetry revealed quasi-reversible electrochemical behavior of the covalently fixed protein with a formal potential of +10 mV vs. Ag/AgCl. Polarized at +150 mV vs. Ag/AgCl the electrode was found to be sensitive to superoxide radicals in the range 300-1200 nmol L-1. Compared with metal needle electrodes sensitivity and reproducibility could be improved and combined with the easiness of preparation. This allows the fabrication of disposable sensors for nanomolar superoxide concentrations. By changing the electrode potential the sensor can be switched from response to superoxide radicals to hydrogen peroxide-another reactive oxygen species. H2O2 sensitivity can be provided in the range 10-1000 mumol L-1 which makes the electrode suitable for oxidative stress studies Y1 - 2004 ER - TY - JOUR A1 - Beissenhirtz, Moritz Karl A1 - Kwan, R. C. H. A1 - Ko, K. M. A1 - Renneberg, Reinhard A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - Comparing in vitro electrochemical measurement of superoxide scavenging activity with an in vivo assessment of antioxidant potential in Chinese tonifying herbs N2 - The in vitro superoxide scavenging activity (as determined by electrochemical measurement) and the in vivo antioxidant potential (as determined by a mouse model of carbon tetrachloride (CCl4) hepatotoxicity) of methanolic extracts prepared from 10 Chinese tonifying herbs were compared. Electrochemical measurement using a cytochrome c (Cyt. c) sensor showed that all of the tested herbal extracts exhibited a medium superoxide scavenging activity of different potency, as indicated by their IC50 values. The in vivo measurement demonstrated that 80% of the herbal extracts displayed in vivo antioxidant potential, as assessed by the percentage of protection of the activity of plasma alanine aminotransferases and the hepatic glutathione regeneration capacity under CCl4-intoxicated condition. Although the in vitro antioxidant activity did not correlate quantitatively with the in vivo antioxidant potential, for 8 out of 10 samples a similar tendency was found. The rapid amperometric assessment of antioxidant potential by Cyt. c sensor may offer a convenient and direct method for screening as well as the quality control of herbal products. Copyright (C) 2004 John Wiley Sons, Ltd Y1 - 2004 ER - TY - JOUR A1 - Beissenhirtz, Moritz Karl A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - A superoxide sensor based on a multilayer cytochrome c electrode N2 - A novel multilayer cytochrome c electrode for the quantification of superoxide radical concentrations is introduced. The electrode consists of alternating layers of cytochrome c and poly(aniline(sulfonic acid)) on a gold wire electrode. The formation of multilayer structures was proven by SPR experiments. Assemblies with 2-15 protein layers showed electrochemical communication with the gold electrode. For every additional layer, a substantial increase in electrochemically active cytochrome c (cyt. c) was found. For electrodes of more than 10 layers, the increase was more than 1 order of magnitude as compared to monolayer electrode systems. Thermodynamic and kinetic parameters of the electrodes were characterized. The mechanism of electron transfer within the multilayer assembly was studied, with results suggesting a protein-protein electron-transfer model. Electrodes of 2-15 layers were applied to the in vitro quantification of enzymatically generated superoxide, showing superior sensitivity as compared to a monolayer-based sensor. An electrode with 6 cyt. c/PASA layers showed the highest sensitivity of the systems studied, giving an increase in sensitivity of half an order of magnitude versus the that of the monolayer electrode. The stability of the system was optimized using thermal treatment, resulting in no loss in sensor signal or protein loading after 10 successive measurements or 2 days of storage Y1 - 2004 SN - 0003-2700 ER - TY - JOUR A1 - Krylov, Andrey V. A1 - Pfeil, Wolfgang A1 - Lisdat, Fred T1 - Denaturation and renaturation of cytochrome c immobilized on gold electrodes in DMSO-containing buffers N2 - Cytochrome c (cyt c) was immobilized on surface-modified gold electrodes using a self-assembling approach. The resulting cyt c electrode was studied using cyclic voltammetry. Compared to pure phosphate buffer, cyt c electrodes exhibited in DMSO-containing solutions lower oxidation and reduction peak currents, which originated from a decrease in the addressable electro-active amount of the surface-immobilized protein. This is associated with the process of protein denaturation. The denaturation kinetics can be described by a sum of two processes with time constants differing by more than one order of magnitude. The subsequent change of the aqueous/organic medium back to a pure aqueous buffer resulted in a shift of the formal potential to its initial value and a partial recovery of the peak current. This can be attributed to the renaturation of the cyt c. The extent of renaturation depended on the organic solvent/water ratio of the mixture used. The kinetics of protein renaturation were similar to those of the denaturation process. (C) 2004 Elsevier B.V. All rights reserved Y1 - 2004 ER - TY - JOUR A1 - Ge, Bixia A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - Electrochemistry of immobilized CuZnSOD and FeSOD and their interaction with superoxide radicals N2 - Copper, zinc superoxide dismutase (CuZnSOD) from bovine erythrocytes and iron superoxide dismutase from Escherichia coli (FeSOD) were immobilized on 3-mercaptopropionic acid (MPA)-modified gold electrodes, respectively. The characterization of the SOD electrodes showed a quasi-reversible, electrochemical redox behavior with a formal potential of 47 ñ 4 mV and -154 ñ 5 mV (vs. Ag/AgCl, 1 M KCl) for surface adsorbed CuZnSOD and FeSOD, respectively. The heterogeneous electron transfer rate constants were determined to be about 65 and 35/s, respectively. Covalent fixation of both SODs was also feasible with only slight changes in the formal potential. The interaction of superoxide radicals (O2-) with the SOD electrode was investigated. No catalytic current could be observed. However, due to the fast cyclic reaction of SOD with superoxide, the communication of the protein with the electrode was strongly influenced. The amperometric detection of superoxide radicals is discussed. Y1 - 2003 ER - TY - JOUR A1 - Beissenhirtz, Moritz Karl A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - Immobilized cytochrome c sensor in organic / aqueous media for the characterization of hydrophilic and hydrophobic antioxidants Y1 - 2003 ER - TY - JOUR A1 - Büttemeyer, R. A1 - Philipp, A. W A1 - Schlenzka, L. A1 - Mall, J. W. A1 - Beissenhirtz, Moritz Karl A1 - Lisdat, Fred T1 - Epigallocatechin gallate can significantly decrease free oxygen radicals in the reperfusion insury in vivo Y1 - 2003 ER - TY - JOUR A1 - Ignatov, S. A1 - Shishniashvili, D. A1 - Ge, Bixia A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - Amperometric biosensor based on a functionalized gold electrode for the detection of antioxidants Y1 - 2002 ER - TY - JOUR A1 - Buttermeyer, R. A1 - Philipp, A. W. A1 - Mall, J. W. A1 - Ge, Bixia A1 - Scheller, Frieder W. A1 - Lisdat, Fred T1 - In vivo measurement of oxygen derived free radicals during reperfusion injury Y1 - 2002 ER - TY - JOUR A1 - Ge, Bixia A1 - Lisdat, Fred T1 - Superoxide sensor based on cytochrome c immobilized on mixed-thiol SAM with a new calibration method Y1 - 2002 ER - TY - JOUR A1 - Scheller, Frieder W. A1 - Wollenberger, Ursula A1 - Lei, Chenghong A1 - Jin, Wen A1 - Ge, Bixia A1 - Lehmann, Claudia A1 - Lisdat, Fred A1 - Fridman, Vadim T1 - Bioelectrocatalysis by redox enzymes at modified electrodes Y1 - 2002 UR - www.elsevier.nl/inca/publications/6/0/1/3/4/7/index.htt ER - TY - JOUR A1 - Fandrich, Artur A1 - Buller, Jens A1 - Memczak, Henry A1 - Stoecklein, W. A1 - Hinrichs, K. A1 - Wischerhoff, E. A1 - Schulz, B. A1 - Laschewsky, André A1 - Lisdat, Fred T1 - Responsive Polymer-Electrode Interface-Study of its Thermo- and pH-Sensitivity and the Influence of Peptide Coupling JF - Electrochimica acta : the journal of the International Society of Electrochemistry (ISE) N2 - This study introduces a thermally responsive, polymer-based electrode system. The key component is a surface-attached, temperature-responsive poly(oligoethylene glycol) methacrylate (poly(OEGMA)) type polymer bearing photoreactive benzophenone and carboxy groups containing side chains. The responsive behavior of the polymer in aqueous media has been investigated by turbidimetry measurements. Polymer films are formed on gold substrates by means of the photoreactive 2(dicyclohexylphosphino)benzophenone (DPBP) through photocrosslinking. The electrochemical behavior of the resulting polymer-substrate interface has been investigated in buffered [Fe(CN)6](3-)/[Fe (CN)6](4-)solutions at room temperature and under temperature variation by cyclic voltammetry (CV). The CV experiments show that with increasing temperature structural changes of the polymer layer occur, which alter the output of the electrochemical measurement. Repeated heating/cooling cycles analyzed by CV measurements and pH changes analyzed by quartz crystal microbalance with dissipation monitoring (QCM-D) reveal the reversible nature of the restructuring process. The immobilized films are further modified by covalent coupling of two small biomolecules - a hydrophobic peptide and a more hydrophilic one. These attached components influence the hydrophobicity of the layer in a different way the resulting change of the temperature-caused behavior has been studied by CV indicating a different state of the polymer after coupling of the hydrophobic peptide. KW - Stimuli-responsive materials KW - electroanalysis KW - modified electrode KW - bioreceptors KW - peptides KW - surface modification KW - cyclic voltammetry KW - IR ellipsometry KW - quartz crystal microbalance Y1 - 2017 U6 - https://doi.org/10.1016/j.electacta.2017.01.080 SN - 0013-4686 SN - 1873-3859 VL - 229 SP - 325 EP - 333 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Riedel, M. A1 - Sabir, N. A1 - Scheller, Frieder W. A1 - Parak, Wolfgang J. A1 - Lisdat, Fred T1 - Connecting quantum dots with enzymes BT - mediator-based approaches for the light-directed read-out of glucose and fructose oxidation JF - Nanoscale N2 - The combination of the biocatalytic features of enzymes with the unique physical properties of nanoparticles in a biohybrid system provides a promising approach for the development of advanced bioelectrocatalytic devices. This study describes the construction of photoelectrochemical signal chains based on CdSe/ZnS quantum dot (QD) modified gold electrodes as light switchable elements, and low molecular weight redox molecules for the combination with different biocatalysts. Photoelectrochemical and photoluminescence experiments verify that electron transfer can be achieved between the redox molecules hexacyanoferrate and ferrocene, and the QDs under illumination. Since for both redox mediators a concentration dependent photocurrent change has been found, light switchable enzymatic signal chains are built up with fructose dehydrogenase (FDH) and pyrroloquinoline quinone-dependent glucose dehydrogenase ((PQQ) GDH) for the detection of sugars. After immobilization of the enzymes at the QD electrode the biocatalytic oxidation of the substrates can be followed by conversion of the redox mediator in solution and subsequent detection at the QD electrode. Furthermore, (PQQ) GDH has been assembled together with ferrocenecarboxylic acid on top of the QD electrode for the construction of a funtional biohybrid architecture, showing that electron transfer can be realized from the enzyme over the redox mediator to the QDs and subsequently to the electrode in a completely immobilized fashion. The results obtained here do not only provide the basis for light-switchable biosensing and bioelectrocatalytic applications, but may also open the way for self-driven point-of-care systems by combination with solar cell approaches (power generation at the QD electrode by enzymatic substrate consumption). Y1 - 2017 U6 - https://doi.org/10.1039/c7nr00091j SN - 2040-3364 SN - 2040-3372 VL - 9 SP - 2814 EP - 2823 PB - Royal Society of Chemistry CY - Cambridge ER -