EP0598095A1 - Mediateurs destines a des enzymes du type oxydoreductases - Google Patents

Mediateurs destines a des enzymes du type oxydoreductases

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Publication number
EP0598095A1
EP0598095A1 EP93913284A EP93913284A EP0598095A1 EP 0598095 A1 EP0598095 A1 EP 0598095A1 EP 93913284 A EP93913284 A EP 93913284A EP 93913284 A EP93913284 A EP 93913284A EP 0598095 A1 EP0598095 A1 EP 0598095A1
Authority
EP
European Patent Office
Prior art keywords
ligand
compound
transition metal
group
sandwich
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP93913284A
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German (de)
English (en)
Inventor
Nigel Forrow
Stephen Walters
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Medisense Inc
Original Assignee
Medisense Inc
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Filing date
Publication date
Application filed by Medisense Inc filed Critical Medisense Inc
Publication of EP0598095A1 publication Critical patent/EP0598095A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • C12Q1/004—Enzyme electrodes mediator-assisted

Definitions

  • the present invention relates to mediators to oxidoreductase enzymes.
  • the present invention relates to such mediators for use m amperometric enzyme electrodes.
  • Amperometric enzyme electrodes are described, for example, in BIOSENSORS Fundamentals and Applications, edited by Turner, Karube and Wilson, Oxford University Press (1987). The reader is referred to this book for background reading.
  • Electrode communication between an electrode and the active site of a redox enzyme is a key feature of amperometric enzyme electrodes.
  • this has been achieved using low molecular weight, rapidly diffusing mediators with fast redox couples.
  • Organic compounds such as phenazine methosulphate, 2,6-dichloroindophenol, quinones and phenylene diamines were among the first such species to be used in conjunction with glucose oxidase (GOD).
  • GOD glucose oxidase
  • these compounds suffer from a number of problems which include poor stability and pH-dependent redox potentials.
  • inorganic mediators suffer from the problem that it is not easy to modify their redox potentials and/or aqueou solubility.
  • the field of inorganic mediators has been ext_ ;>' d by ferrocenes, which are sandwich compounds with an iron atom sandwiched between two ⁇ -bonding cyclopentadienyl rings.
  • the access to a large number of derivatives means that the properties of ferrocenes can readily be modified by substitution on one or both of the cyclopentadienyl rings.
  • Ferrocene mediators have been shown to undergo rapid electron transfer with oxidoreductase enzymes such as glucose oxidase (Anal. Chem., 56
  • a principal object of the present invention is the provision of compounds of use as mediators for oxidoreductase enzymes or other electron transfer redox proteins.
  • Related objects include mediation processes employing such mediators, products based on the mediation processes, manufacturing methods for the mediated products, and compositions containing the mediators.
  • ⁇ -ligand which may be allyl, cyclopropenyl, cyclobutadiene, cyclopentadienyl, benzene, cycloheptatrienyl or trimethylenemethane, among others
  • oxidoreductase enzymes such as glucose oxidase
  • the mediating function is not restricted to enzymes, and may be used with other electron transfer redox proteins.
  • the present invention provides transition metal half-sandwich complexes, for use as mediators in electron transfer between enzymes and electrodes.
  • the present invention provides a process for mediating electron transfer between an oxidoreductase enzyme and an electrode, which is characterised by the use of a transition metal half- sandwich compound as mediator.
  • a transition metal half- sandwich compound as mediator.
  • the present invention provides mediated enzyme products and methods of manufacturing such products.
  • the present complexes can be used in the manufacture of amperometric enzyme sensors, such as those of the kind described in EP-78636-A, EP 125136- A, EP 125137-A, EP 125139-A, EP 125867-A, EP 127958-A or EP 230786-A, but using a transition metal half-sandwich compound as mediator.
  • the present invention provides amperometric enzyme sensors of use for testing for the presence of an analyte in a sample, especially an aqueous sample.
  • the sample can be a complex biological sample, for instance a biological fluid such as whole blood, plasma or serum, and the analyte can be a naturally occurring metabolite such as glucose or cholesterol, or an introduced substance such as a drug.
  • the present invention further provides an electrically conductive printing ink which includes a transition metal half-sandwich mediator compound.
  • a central feature of the invention is the use as a redox mediator for oxidoreductase enzymes of a half-sandwich complex where a single ⁇ -ligand is bonded to a transition metal ion complexed by other ligands.
  • the single ⁇ -ligand will comprise 3 or more atoms, typically 3 to 7 atoms, which will generally be carbon but can include any heteroatom capable of ⁇ bonding including nitrogen, oxygen, sulphur and phosphorus.
  • at least three interlinked atoms of the ⁇ -ligand define a plane with delocalised ⁇ bonding and which interact directly with the transition metal atom.
  • transition metal ion is not critical, except for the ability to undergo rapid electron transfer.
  • identity of the other ligands is not critical, except that they are not ⁇ -bonding.
  • the other ligands may be chosen from any of carbonyl, phosphine, isocyanide, nitrosyl, bipyridine, phenanthroline or dithiolate.
  • the half-sandwich compound for use as mediator is a complex of the general formula:
  • ( ⁇ -ligand)M(not- ⁇ -ligand) n wherein the complex may be charged or neutral, ⁇ -ligand represents the single ⁇ -bonded ligand, represents the transition metal atom, n is the number of ligands which are not ⁇ -ligands, and the n not- ⁇ -ligands are the same or different, are univalent or multivalent, and serve to satisfy the valency of the transitional metal M.
  • the ⁇ -ligand is selected from ⁇ -ligands with 3 to 7 carbon atoms, and is most preferably an allyl, cyclopropenyl, cyclobutadiene, cyclopentadienyl, benzene, cycloheptatrienyl or trimethylenemethane group.
  • the ⁇ -ligand can be unsubstituted, but is preferably a substituted ligand.
  • the nature of the substituent is not critical, since it is a feature of the present invention that variation in the identity of the substituent allows the preparation of compounds with different properties for assessment as candidate mediators. Purely by way of illustration, there can be 1 or more, usually 1 to 4, substituents.
  • Such substituents may be selected from alkyl groups, especially a methyl or ethyl group; alkoxy groups, especially a methoxy or ethoxy group; amino and substituted amino groups, especially an amino, dimethylamino, diethylamino or di( ⁇ -hy ⁇ _roxyethyl)amino group; aminoalkyl and substituted aminoalkyl groups, especially an aminomethyl or dimethylaminomethyl group; a carboxylic acid group; or a suiphonic acid group.
  • the transition metal M is preferably iron, manganese or chromium.
  • Each not- ⁇ -ligand is preferably selected from carbonyl, phosphine, isocyanide, nitrosyl, bipyridine, phenanthroline or dithiolate groups.
  • the transition metal half-sandwich compound is employed as an inclusion complex in ⁇ -cyclodextrin, which typically takes the form of a 1 : 1 adduct.
  • the transition metal half-sandwich compounds are generally known or can be made by modified literature methods.
  • transition metal half-sandwich complexes over the ferrocenes is that the greater degree of structural possibilities allows their properties to be tailored more closely to fit a particular enzyme application and/or biosensor configuration.
  • Other applications and advantages of the present invention will be apparent, such as the possibility of the use of transition metal half-sandwich complexes as mediators to oxidoreductase enzymes, such as the generality of the flavoprotein, metalloprotein and quinoprotein oxidoreductases; also to act as mediators to redox proteins such as haemoglobin or to respiratory or other electron transfer redox proteins; the possibility of their use as labels for antigens or antibodies in immunochemical procedures including such as described in EP 125139-A; the possibility of their use as artificial substrates to monitor enzyme reactions with redox potentials modified by reaction with the enzyme; the possibility of tailoring the redox potential of a mediator by careful choice of ligands such that it may interact with a particular oxidoreductase protein but not with
  • the preferred amperometric enzyme sensor adopting a transition metal half- sandwich compound as mediator utilizes a strip test element, especially a throw-away dry strip.
  • a disposable test element carries a working electrode, the test reagents comprising at least the oxidoreductase enzyme and transition metal half-sandwich mediator compound for generation of a current indicative of the level of analyte, and a reference electrode.
  • the test reagents are in one or more carbon-based layers associated with the working electrode.
  • the sensor electrodes preferably comprise electrode areas formed for instance by screen printing, spraying, or other suitable deposition technique.
  • the present invention provides a dry strip sensor which comprises an elongate, electrically-insulating carrier having a pair of longitudinal, substantially parallel electrically-conducting tracks thereupon, each track being provided at the same end with i.. « ⁇ ans for electrical connection to a read-out means and provided with an electrode, one of the electrodes being the reference electrode and the other being the working electrode, together with test reagents.
  • such a sensor is configured in the form of a supporting strip of electrically insulating carrier material such as a synthetic polymer (for instance pvc) carrying at a location between its ends the two electrodes supported on electrically conductive printed tracks.
  • the electrodes can take the form of two rectangular areas side by side on the carrier strip. Such areas can be configured as a target area to be covered by a single drop of sample, such as whole blood, for testing for an analyte.
  • non-rectangular electrode areas for instance diamond-shaped, semicircular, or triangular areas, can be employed to provide a target area for optimised contact by a liquid sample.
  • the carrier carries at least two electrodes, namely a reference electrode and a working electrode, but may carry further electrodes such as a counter electrode and/or a dummy electrode, which is of similar formulation to the working electrode with associated test reagents, but lacks one or more of its active components.
  • a dummy third electrode can lead to more reliable results, in that if charge passed at the third electrode is subtracted from charge passed at the working electrode, then the resulting charge is solely due to the reaction of interest.
  • a membrane may be provided at and above the target to perform a filtration function, for example to filter blood cells from the sample before it enters the test strip.
  • a filtration or cellular separation membrane might be cast in situ.
  • a single use disposable electrode strip for attachment to signal readout circuitry of a sensor system to detect a current representative of a compound in a liquid mixture
  • the strip comprising: a) an elongated support adapted for releasable attachment to the readout circuitry; b) a first conductor extending along the support and comprising a conductive element for connection to the readout circuitry; c) an active electrode on the strip in contact with the first conductor and positioned to contact the mixture, the active electrode comprising a printed layer on the support, the printed layer comprising an enzyme capable of catalyzing a reaction involving a substrate for the enzyme and a transition metal half-sandwich mediator capable of transferring electrons transferred between the enzyme-catalyzed reaction and the first conductor to create a current representative of the activity of the enzyme and representative of the compound, the printed layer having an area less than 25 square mm; d) a second conductor extending along the support, comprising a conductive element
  • test strips of this invention can detect analytes for example by known mediated enzyme electrochemical reactions, but using a transition metal half-sandwich compound as mediator.
  • the reader is referred to European Patent 78636-A, especially for its mention of glucose oxidase, as well as European Patent 127958-A and European Patent Specification 125137-A.
  • mediators for use with the present mediators include combinations of cholesterol oxidase, a peroxidase and optionally cholesterol esterase, for detection of free or total cholesterol, as described in European Patent 230786-A; and more generally systems including oxidoreductase or other enzymes, including flavoproteins, quinoproteins, and metalloproteins, and one or more mediator compounds, for example as described in European Patent Specification 125137-A.
  • enzyme systems suitable for use in devices according to the present invention include oxidoreductases using nicotinamide and adenine dinucleotide and nicotinamide adenine dinucleotide phosphate as cofactors, for example those described in European Patent 125867-A; or systems using more than one enzyme, as described in European Patent 125136- A.
  • Test strips according to this invention are intended for use with electronic apparatus and meter systems. These control the progress of the electrochemical reaction, for example by maintaining a particular potential at the electrodes, and monitor the reaction and calculate and present the result.
  • a particular feature which is desirable in a meter system for use with test strips of this type is the capability to detect the wetting of the reaction zone by sample fluid; this allows timely initiation of the measurement and reduces the scope for inaccuracies caused by user error. It is possible to achieve this goal by applying a potential to the electrodes of the test strip as soon as it is inserted into the meter; this potential is maintained until a certain threshold current is measured which will reflect the wetting of the reaction zone by sample fluid. The potential may then be removed for a short time to allow wetting to V completed before initiation of measurement in the usual way.
  • a further feature which is advantageous is the facility to ascertain automatically the identity of test strips for the measurement of different analytes. This might be achieved, for example, by the provision of additional contacts on the test strip in a pattern characterising the type of strip; the meter would have contact pads to determine the presence or absence of each contact.
  • a system is possible using one or more additional circuit loops printed on the test strip of determined resistance characteristic of the type of strip, as described in US 5126034 at column 4, lines 3 to 17.
  • notches or other shapes might be made in the proximal end of the test strip which is inserted into the meter; switches or optical detectors in the meter can detect the presence or absence of each notch.
  • Further possibilities for recognition of strip types include varying the colour of the strips and providing the meter with a photodetector capable of distinguishing the range of colours; also providing the strips with barcodes; magnetic stripes or other markings and providing the meter with a suitable reading arrangement.
  • the strip electrodes have a three-electrode configuration comprising a reference electrode, a working electrode, and a dummy electrode.
  • the carrier can be made from any material which has an electrically insulating surface including poly(vinyl chloride), polycarbonate, polyester, paper, cardboard, ceramic, metal coated with ceramic or other insulating substance.
  • poly(vinyl chloride) or a blend of polycarbonate/polyester seems to be particularly suitable.
  • a sheet of carrier material is cut to size using a punch tool which crops the outer edges at the same time as punching a number of registration holes. It is advantageous to use these holes during subsequent operations, but it is also feasible to edge-register throughout the process.
  • a conductive ink is applied to the carrier by a method of deposition such as screen printing.
  • the punched holes are used to register the carrier.
  • This layer produces the contact areas which allow a meter to interface with the test strip and provides an electrical circuit between the contacts and the active chemistry occurring on the strip.
  • the ink is an organic solvent-based carbon mixture which is air-dried.
  • Alternative formulations are possible including water-based inks and silver inks. Other methods of drying or curing the inks include infra-red, ultra-violet and radio- frequency radiation.
  • a layer forming the reference electrode is then printed with an organic solvent-based ink containing a silver/silver chloride mixture.
  • the print extends to partially cover the middle track of the carbon print where it extends into the reaction zone. It is useful if separate parts of this print are extended to cover parts of the other carbon tracks outside the reaction zone so that the total electrical resistance of each track is reduced.
  • a layer of dielectric ink may optionally be printed to cover the majority of the printed carbon and silver/silver chloride layers. In this case, two areas are left uncovered, namely the electrical contact areas and the sensing area which will underlie the reactive zone. This print serves to define the area of the reactive zone, and to protect exposed tracks from short circuit.
  • one or more inks are deposited to a precise thickness within a defined area on top of one of the conductive tracks within the reaction zone, to deposit the enzyme and the half-sandwich mediator compound. It is convenient to do this by means of screen printing. Other ways of laying down this ink include inkjet, volumetric dosing, gravure printing, flexographic printing, and letterpress printing.
  • a second partially active ink forming a dummy electrode can be deposited on a remaining conductive track, such that each active ink lies on a conducting track equidistant from a central track coated with the silver/silver chloride ink.
  • the phosphate buffer solution (PBS) of 0.15 M phosphate (with 0.2 M sodium chloride as supporting electrolyte), pH 7.0, containing 0.1 M glucose was prepared using Analar reagents from Merck BDH. The solution was stored overnight at 4°C prior to use to allow equilibration of the - and ⁇ - anomers.
  • Glucose oxidase (EC 1.1.3.4 from Aspergillus niger) was supplied by Sturge Enzymes. Horseradish peroxidase was obtained from Biozyme.
  • Figure 1 is a cyclic voltammogram of the chromium complex Compound 6.
  • Figure 2 comprises (a) cyclic voltammograms of electrodes containing the chromium complex Compound 6, and (b) the catalytic wave upon addition of glucose oxidase.
  • Figure 3 shows catalytic waves obtained on titrating glucose oxidase with the manganese complex Compound 8.
  • Figure 4 is a graphical plot of data obtained from the catalytic waves of Figure 3.
  • Figure 5 is a cyclic voltammogram of the manganese complex Compound 16.
  • Figure 6 comprises (a) a cyclic voltammogram of the ⁇ -cyclodextrin adduct of the manganese complex Compound 10, and (b) the catalytic wave upon addition of glucose oxidase.
  • Figure 7 comprises cyclic voltammograms of the manganese complex Compound 16 in the presence of different amounts of ⁇ -cyclodextrin.
  • Figure 8 comprises cyclic voltammograms of electrodes containing the manganese complexes (a) Compound 10, (b) Compound 1 1 , and (c) Compound 12.
  • Figure 9 comprises (a) cyclic voltammograms of electrodes containing the manganese complex Compound 10, and (b) the catalytic wave upon addition of glucose oxidase.
  • Figure 10 shows the response to glucose of poised glucose oxidase/Compound 10 electrodes.
  • Example 2 The half wave potential of benzene chromium tricarbonyl has been recorded as +710mV vs SCE in dichloromethane (J. Chem Soc. Dalton Trans., (1973) 1768). This complex has negligible solubility in buffer. Derivatives, however, are simple to prepare in good yield using the thermolysis of chromium hexacarbonyl with substituted benzenes in tetrahydrofuran/di-n- butyl ether (Inorg. Synth., 19 (1979) 154). Arenes containing amines directly attached to the ring were selected in the hope of enhancing the aqueous solubility of the resulting complexes as well as reducing their mid ⁇ point potentials. A series of seven complexes were investigated and were found to demonstrate aqueous electrochemistry in the region +280 to +40mV, as shown in the following table.
  • Figure 1 is a cyclic voltammogram of th , . romium complex Compound 6, at 50 mV/s in PBS.
  • Compound 6 was - . only compound to display a reversible wave, as shown Figure 1, though it was poorly soluble in aqueous buffer. Further studies were confined to Compound 6.
  • FIG. 2 comprises (a) cyclic voltammograms of electrodes containing the chromium complex Compound 6 at 50 mV/s and 0.2 uA/cm in PBS, and (b) the catalytic wave upon addition of 100 ⁇ l glucose oxidase recorded at 20 mV/s and 0.5 ⁇ A/cm. It will be seen from Figure 2(a) that a reversible wave was recorded when PBS was applied to the electrodes, but at a lower mid-point potential of - 15 mV. From Figure 2(b), it is seen that GOD mediation occurs on applying mixtures of enzyme and glucose to the electrodes, with small catalytic currents of about 1 ⁇ A.
  • Manganese half-sandwich complexes can be obtained from (methylcyclopentadienyl)manganese tricarbonyl Compound 8, an inexpensive and readily available starting material due to its large scale use as an anti-knocking additive in unleaded petrol.
  • access to a wide and varied carbonyl substitution chemistry is provided via the nitrosyl cation Compound 9 produced by treatment of the tricarbonyl compound Compound 8 with nitrous acid (Inorg. Chem., 6 (1967) 469) or nitrosonium ion (J. Organomet. Chem, 56 (1973) 345).
  • Cyclopentadienyl manganese tricarbonyl itself is more difficult to oxidise than ferrocene; its oxidation potential being some 1.2 V more positive than ferrocene in acetonitrile (J. Organomet. Chem., 101 (1975) C43).
  • the nitrosyl cation Compound 9 readily decomposes in aqueous buffer to give a species which is irreversibly oxidized at +230 mV.
  • treatment of Compound 8 with cyanide ion (J. Organomet. Chem., 170 (1979) 217) affords the anionic dicyano complex Compound 13 which is highly water soluble and displays a reversible wave at +440 mV.
  • Figure 3 shows catalytic waves obtained at 5 mV/s and 5 ⁇ A/cm on titrating glucose oxidase with the manganese complex Compound 8, using 0 to 25 ⁇ l GOD titrated into a solution of Compound 8 in 0.1 M glucose/PBS.
  • a series of catalytic waves of increasing amplitude were obtained.
  • a second order rate coefficient for the oxidation of reduced GOD of 2.1 x 10-5 M ⁇ ls ⁇ l was derived from the graph shown in Figure 4 of i ca t versus [GOD] 1 / 2 plotted according to literature methods (Prog. React. Kinet., 16 (1991) 55, and J. Electroanal. Chem., 293 (1990) 55).
  • Compound 16 was prepared from cis-l,2-dicyanoethylene-l,2-dithiolate (Inorg. Chem., 8 (1969) 1340).
  • Figure 5 is a cyclic voltammogram of the manganese complex Compound 16 0.6 mM in PBS at 50 mV/s and 2 ⁇ A/cm. A much lower E ox of -25 mV was observed for the anionic dithiolene complex, but the oxidized form (a neutral species) proved to be insoluble in buffer. The resultant coating of the electrode meant that an enzyme titration was not possible.
  • Figure 6 comprises (a) a cyclic voltammogram of the ⁇ -cyclodextrin adduct of the manganese complex Compound 10 in 0.1 M glucose/PBS, and (b) the catalytic wave upon addition of 50 ⁇ l glucose oxidase.
  • ⁇ -cd is capable of hosting a neutral manganese half sandwich complex
  • ⁇ -cd might act as a solubilising agent for the insoluble oxidized form of the anionic complex Compound 16 mentioned earlier.
  • the cyclic voltammogram of a 0.5 mM solution of Compound 16 in buffer has features which indicate that a species is being precipitated onto the electrode.
  • addition of ⁇ -cd (0.5- 5.0mM) resulted in a reversible wave.
  • Figure 7 comprises cyclic voltammograms of the manganese complex Compound 16 in the presence of different amounts of (a) PBS only, and PBS containing (b) 0.5 mM ⁇ -cd, (c) 1.0 mM ⁇ -cd, and (d) 5.0 mM ⁇ -cd.
  • mediator/enzyme electrodes responded linearly as shown in Figure 10 to glucose in the range 0 to 12 mM when poised at +450 mV. Inclusion of ethylene glycol in the formulation of the carbon ink containing enzyme gave currents of up to 190 ⁇ A with 0.1 M glucose.
  • HRP horseradish peroxide
  • the catalytic ring current was then measured while poising the ring electrode at -100 mV and scanning the disk electrode from -100 to - 600m V to generate hydrogen peroxide. Successive experiments with the ferrocene and the ⁇ -cd adduct of Compound 10 resulted in identical catalytic currents being recorded.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
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  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Parts Printed On Printed Circuit Boards (AREA)
  • Inks, Pencil-Leads, Or Crayons (AREA)
  • Conductive Materials (AREA)

Abstract

Cette invention concerne des complexes semi-sandwichs de métaux de transition comprenant un pi-ligand unique comportant une liaison pi délocalisée qu'on utilise comme médiateurs de transfert d'électrons entre une électrode et une protéine redox, par exemple une enzyme du type oxydoréductase telle que celle existant dans un capteur enzymatique ampérométrique. Les composés appropriés comprennent des complexes de formule générale: pi-ligand)M(non-pi-ligand)n. Dans cette formule, le complexe est chargé ou neutre, pi-ligand représente le ligand unique à liaison pi, M représente l'atome de métal de transition, n représente le nombre de ligands qui ne sont pas des pi-ligands, et les n non-pi-ligands sont identiques ou différents, univalents ou multivalents, et servent à satisfaire la valence du métal de transition M.
EP93913284A 1992-06-05 1993-06-03 Mediateurs destines a des enzymes du type oxydoreductases Withdrawn EP0598095A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GB9212010 1992-06-05
GB929212010A GB9212010D0 (en) 1992-06-05 1992-06-05 Mediators to oxidoreductase enzymes
PCT/GB1993/001179 WO1993025898A1 (fr) 1992-06-05 1993-06-03 Mediateurs destines a des enzymes du type oxydoreductases

Publications (1)

Publication Number Publication Date
EP0598095A1 true EP0598095A1 (fr) 1994-05-25

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EP93913284A Withdrawn EP0598095A1 (fr) 1992-06-05 1993-06-03 Mediateurs destines a des enzymes du type oxydoreductases

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EP (1) EP0598095A1 (fr)
JP (1) JP3288044B2 (fr)
CA (1) CA2115052A1 (fr)
GB (1) GB9212010D0 (fr)
WO (1) WO1993025898A1 (fr)

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CA2115052A1 (fr) 1993-12-23

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