WO1995020051A1 - Detecteurs proptochimiques dont le fonctionnement est fonde sur l'inhibition enzymatique - Google Patents

Detecteurs proptochimiques dont le fonctionnement est fonde sur l'inhibition enzymatique Download PDF

Info

Publication number
WO1995020051A1
WO1995020051A1 PCT/US1995/000846 US9500846W WO9520051A1 WO 1995020051 A1 WO1995020051 A1 WO 1995020051A1 US 9500846 W US9500846 W US 9500846W WO 9520051 A1 WO9520051 A1 WO 9520051A1
Authority
WO
WIPO (PCT)
Prior art keywords
species
sensor
substrate
enzyme
optical
Prior art date
Application number
PCT/US1995/000846
Other languages
English (en)
Inventor
Otto S. Wolfbeis
Stanley M. Klainer
Original Assignee
Fci-Fiberchem, Inc.
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Fci-Fiberchem, Inc. filed Critical Fci-Fiberchem, Inc.
Publication of WO1995020051A1 publication Critical patent/WO1995020051A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/84—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving inorganic compounds or pH
    • 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
    • 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
    • C12Q1/58—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving urea or urease

Definitions

  • the invention relates generally to optical chemical sensors and more particularly to enzyme based optical chemical sensors.
  • optical chemical sensors are based on optical fibers and other waveguides.
  • Many optical chemical sensors (OCS) are solid state; others are reservoir type. These sensors generally contain a chemistry which reacts with the target species and produces a measurable optical effect. However, many target species cannot be detected because a suitable reaction does not exist.
  • One approach to solving this problem of not being able to directly measure a target species is to convert the target species to a product which can be measured.
  • a biological transducer e.g., an enzyme, which converts the analyte into a species for which an optrode exists.
  • the complex is adsorbed onto silica gel and incorporated into a silicone matrix with high oxygen permeability placed on the tip of a fiber.
  • the enzyme glucose oxidase is immobilized on the surface of the oxygen optrode.
  • the sensor relates oxygen consumption as a result of enzymatic oxidation of glucose to glucose concentration.
  • an oxygen optrode with an oxygen sensitive indicator dye (decacyclene) and a C0 2 optrode with a pH sensitive indicator dye (HPTS) having the enzymes gluta ate oxidase and glutamate decarboxylase, respectively, immobilized thereon are used to detect L-glutamate, Dremel, et al., "Comparison of two fibre-optic L-glutamate biosensors based on the detection of oxygen or carbon dioxide, ...n, Analytica Chimica Acta, 248(1991)351-359.
  • Microbial sensors e.g. as described in U.S. Patent Applications Ser. No. 08/163,040 filed Dec. 6, 1993 and Ser. No. 08/101,977 filed Aug 4, 1993, contain micro ⁇ organisms, e.g., yeasts or bacteria, immobilized on an optical chemical sensor. The microorganisms act on the target species to produce a measurable change in a reaction product.
  • a target species cannot be directly detected by a suitable reaction and cannot be converted to a detectable product.
  • heavy metals HMs
  • an additional sensor mechanism is needed which does not involve direct measurement of the target species or one of its reaction products.
  • HMs Heavy Metals
  • HMs Optical detection and quantification of HMs is usually performed using spectrophotometric or fluorometric cuvette tests, or with commercially available test strips. While such tests are widely accepted as the state-of the art, they do not provide good sensitivity and selectivity and can, therefore, be used only as an indicator of the existence of a problem.
  • the heavy metals are very similar in behavior and thus there is no single reagent or combination of reagents that allows them to be spectrally separated. Consequently, after selecting the best reagents, algorithms (Chemometrics) must be written to improve separation by the use of software.
  • one objective of the invention is to use the inhibition effect of a target species, e.g., enzyme inhibition by heavy metals, in an optical chemical sensor.
  • Another objective of the invention is to provide the means for a test that can be performed in the field by unskilled personnel and to overcome the problem of varying buffer capacity.
  • This invention preferably addresses the need for: (1) General sensors which can be used for indicating the presence of general groups of analytes, (2) Individual sensors which can detect and quantify a particular species and (3) Group sensitive sensors which can identify and measure particular chemical sets, i.e. heavy metals.
  • the general sensors are not intended to be species specific nor are they intended to be quantitative. They must, however, have part-per-billion (ppb) sensitivity to be useful. They are intended to be early warning devices to provide inputs as to when samples are to be collected and subsequently analyzed in the laboratory. Thus they provide information that a target or group of targets are present and furnish the mechanism for eliminating the analyses associated with negative samples.
  • These sensors can also be used to assure that a target or the sum of several targets remain below a threshold level.
  • the analyte specific devices are intended to be true monitoring devices. Their purpose is to look for a particular species; identify its presence and its concentration insitu and in real-time; assure that the amount present is within predetermined limits; and, if necessary, initiate the proper correction procedures or institute the appropriate warnings.
  • One aspect of the invention is an optical chemical sensor method and apparatus for detecting a target species by its effect in inhibiting the action of an enzyme on another species which produces a detectable product.
  • An enzyme is selected which may act on a chemical substrate to produce a product which is detectable with an available optical chemical sensor, and whose action on the substrate is inhibited by the target species.
  • a wide variety of chemical sensors can preferably be used including waveguide sensors and reservoir sensors. Both general sensors and species-specific sensors can be produced.
  • a heavy metal sensor is based on inhibition of urease which hydrolyzes urea, producing ammonia or ammonium.
  • an optical sensor for detecting a first species comprising: optical sensing means which produces an optical signal which varies as a function of concentration of a second species; a chemical substrate; an enzyme which acts on the chemical substrate to produce the second species and which is inhibited by the first species from acting on the substrate to produce the second species; and a sensor comprising a buffer.
  • an optical method for detecting a first species comprising: exposing a chemical substrate to an agent which acts on the substrate to produce a second species and which is inhibited by the first species from acting on the substrate to produce the second species; optically detecting changes in concentration of the second species; determining changes in the first species from detected changes in the second species.
  • Fig. 1 is a perspective view of a rectangular cuvette according to the invention.
  • Fig. 2a is a perspective view of a triangular cuvette according to the invention.
  • Fig. 2b is a sectional view of a second cuvette containing substrate.
  • Figs. 3a,b illustrate an integrated waveguide capillary cuvette.
  • Fig. 4 is a sectional view of a waveguide sensor.
  • Figs. 5a,b are sectional views of a reservoir sensor.
  • Figs. 6a-c show a multiple waveguide sensor.
  • Fig. 7 is a response curve for an ammonia sensor at various concentrations.
  • Fig. 8 is a response curve for an ammonia sensor as a function of pH.
  • Fig. 9 shows the Michaelis-Menten diagram and the Lineweaver-Burk plot of urease.
  • Fig. 10 shows the inhibition of urease by Ag'.
  • Figs. 11a-j show the inhibition of urease by various heavy metals, when immobilized with and without covalent bonds.
  • the invention is method and apparatus for measurement with an optical waveguide chemical sensor (OWCS) or other optical chemical sensor (OCS) of a target chemical 0051
  • OWCS optical waveguide chemical sensor
  • OCS optical chemical sensor
  • the OWCS or OCS includes a chemical substrate on which the enzyme acts to produce a detectable product.
  • the target species inhibits the activity of the enzyme so that changes in the detectable product are related to the target species.
  • the basic reaction is:
  • a two enzyme system can be used where enzyme B converts Product A to Product B which can be detected. Enzyme A is still inhibited by the target species so a decrease in Product A leads to a decrease in Product B which is thus an indicator of the target species.
  • This invention involves the approach whereby an enzyme reacts with a particular analyte (substrate) to generate a species which can be measured by an optical chemical sensor (OCS) .
  • OCS optical chemical sensor
  • This is the baseline reaction and represents no inhibitor present.
  • the enzyme selected for the baseline reaction is picKed to be inhibited by the target molecule or ion of interest. When this inhibition takes place, the amount of species to be measured by the OCS is reduced and the amount of signal decrease can be related to the concentration of the molecule or ion of interest.
  • the induced fit is a dynamic recognition process, i.e., the molecular conformation change is responsible for specificity and t2)
  • the enzyme introduces an electronic strain in its substrate to expedite enhanced reaction rates. For these reasons, enzymes are often more suitable for initiating specific chemical reactions than reagents and dyes.
  • the use of enzyme inhibition represents a unique method of detecting and quantifying a variety of species, including the heavy metals and several anions, organics and gases.
  • the approach has the general advantages of: (1) using a simple OWCS or OCS for making difficult chemical analyses, (2) doing complex analyses without sample preparation, (3) making real-time in-situ measurements, (4) selecting between reversible and non-reversible reactions, (5) choosing between analyzing for a single analyte or a group of analytes, (6) having good selectivity and sensitivity and (7) being amenable to solid-state sensor configurations.
  • a particular benefit of this approach is that since enzymes can now be tailored to meet any reaction mechanism requirement, the potential for very broad analytical applications for this approach is enhanced.
  • Table 1 gives a limited list of analyses that can be done using the invention.
  • the first column gives the species to be detected and measured
  • the second column a selection of enzymes which are inhibited by the species of interest
  • the third column the OCS which will be used for the measurement.
  • the number of targets that can be measured is only restricted by the available enzymes and OCS. Of particular importance is the limited number of OCS needed to measure a large number of target analytes.
  • detection can be accomplished using an ammonia OWCS.
  • the lyase forms oxaloacetate which in a second enzyme reaction is decarboxylated to form carbon dioxide.
  • the first reaction yields glutamate which, in a second enzyme-catalyzed reaction, is oxidized by glutamate oxidase/oxygen to form amnonia and hydrogen peroxide. 6) Arginase produces urea which is converted, in a second step, to carbon dioxide and amnonia by urease.
  • a competing enzyme system (creatinase and urease) is added to the enzyme (creatine kinase) which is inhibited, (a) If not inhibited, the kinase catalyzes the usual reaction (the formation of phosphocreatine). (b) j_ inhibited, the competing reaction takes place, which is the conversion of creatine into urea, followed by hydrolysis of urea to give ammonia and carbon dioxide.
  • This scheme requires the kinase to have a higher activity than the creatinase.
  • the appropriate substrates for the enzymes there is an obvious substrate for each enzyme, e.g. listed in T.E.
  • enzymes are more general in their action and accept several substrates. Others are very specific; e.g. urease is specific for urea.
  • the most appropriate substrate is usually the one given by the enzyme's name, e.g., glucose for glucose oxidase, acetylcholine for acetylcholine esterase, methane for -methane monooxygenase, creatine for creatine kinase, analine for analine carboxypeptidase and peptides for amino peptidase.
  • optical waveguide covers optical waveguides per se; channeled optical waveguides; single and multimode fiber optics, with either side- or tipcoatings and optical "chips".
  • OWCS optical waveguides
  • the invention can also be carried out with other types of optical chemical sensors, e.g., reservoir type sensors, including simple cuvette systems.
  • the reservoir sensors can include a fiber optic.
  • Another part of this invention is the need to maintain the enzymes in an active state. Enzymes are known to degrade as a function of time, temperature and other parameters which often cannot be controlled during a sensor's use or storage. To overcome these effects, with the exception of extreme temperatures, two steps have been taken: (1) The enzyme is immobilized onto a support, e.g., cell wall. (Preference would have been to covalently bond the enzyme, but this results in lack of participation in the reaction as will be shown later.) Thus, the enzyme, and also substrate and buffer, are deposited or immobilized on the walls of a cell before aqueous sample is added, but in a manner that they readily dissolve when the sample is added.
  • a support e.g., cell wall.
  • Figure 1 is a rectangular cuvette 10 with a sensor chemistry (paint) 12 deposited on one of the inner walls. On another wall substrate 13 and buffer 14 are deposited while on a third wall enzyme 16 is deposited.
  • the sample will dissolve the enzyme, substrate and buffer 12 (but not the sensor chemistry) and the reaction will begin. This results in a liberation of a chemical species such as ammonia.
  • a second cell like cuvette 10 in Figure 1, can be used as a reference cell.
  • the reference cell is filled with plain water or buffer instead of sample and a difference measurement is performed with two beams. One beam measures non-inhibited activity in the reference cell while the second beam measures the inhibited activity in the sample cell.
  • Figure 2a is a triangular cuvette 22 configuration with the sensor chemistry (paint) 12 deposited on one wall.
  • the substrate 13 and buffer 14 are deposited on the bottom of cuvette 22.
  • the enzyme 16 is deposited on a second wall.
  • the sample will dissolve all components (enzyme, substrate, buffer) and the reaction will begin. This results in a liberation of a chemical species such as oxygen or carbon dioxide.
  • the excitation light 1 ⁇ causes fluorescence 1 ⁇ to occur.
  • the fluorescence intensity 1 ⁇ is a measurement of the amount of reaction taking place and thus the analyte concentration.
  • a 2- ⁇ ell configuration can be produced by the combination of Figures 2a and 2b.
  • a triangular cuvette 22 has sensor chemistry (paint) 12 deposited on one wall.
  • a second wall is coated with enzyme 16 and buffer 14.
  • the sample containing the inhibitor would be added to a second container, cuvette 28 of Fig. 2b, of known volume containing the solid substrate 30.
  • the contents of cuvette 28 would then be poured into cuvette 22 and the reaction measured.
  • the cell 22 is filled with an aqueous sample, the sample will dissolve all components and the reaction will begin. This results in a liberation of a chemical species. If ammonia or carbon dioxide is to be assessed, the measurement will be made in absorption or fluorescence; while if oxygen is to be assessed, fluorescence is the detection method.
  • FIGS 3a and 3b show two views, assembled and partly disassembled, of, an "integrated waveguide capillary cuvette” (IWCC) 32.
  • IWCC 32 is formed of body 20 with cavity 24 formed therein and capillary inlet 26.
  • the sensor chemistry (paint) 12 is on the inside surface of one side 21 of the IWCC and the immobilized enzyme 16 is on the inside surface of the other side 23.
  • the sample containing the inhibitor would be added to a second container, cuvette 28 of Fig 2b, of known volume containing the solid substrate. The contents of cuvette 28 would then enter IWCC 32 by capillary action through inlet 26.
  • UN is input into waveguide 33 and changes in the light signal propagating through the waveguide 33 are detected.
  • Figure 5a shows a fiber optic reservoir cell 50 (Klainer, et al, U.S. Patent No. 5,059,790, U.S. Patent No. 5,107,133 and U.S. Patent No. 5,116,759) adapted to the present invention.
  • a fiber optic consisting of core 42 and clad 44 is placed in a reservoir cell of known volume 46.
  • Sensor chemistry 12 is placed on the tip of the fiber.
  • One wall of the cell is coated with substrate 13 and buffer 14.
  • a second wall is coated with enzyme 16.
  • the sample to be measured enters through inlet 52 15 and exits through outlet 54. This approach is suitable for use with kinetic and static samples.
  • Figure 5b is a variation of Figure 5a wherein part of clad 44 is removed and replaced with sensor chemistry (paint) 12, i.e., the sensor chemistry is on the side (side-coated) rather than the tip.
  • sensor chemistry paint
  • Figures 6a-c show a sensor 58 with multiple waveguides on a chip 56.
  • three different waveguides 60, 62 and 64 are used. This permits three different enzymes to be used at once and also provides for a reference channel 66.
  • the system uses a single light source 68 and multiple detectors 70, 72 and 74 each of which is filtered to give a specific detection wavelength.
  • a fourth detector 76 can be used to look at the reference channel 66.
  • Light from source 68 in chip 56 is incident on sloped reflective end face 61 of waveguide 59 which reflects the light down the waveguide 59 to beam splitter 71. Beam splitter 71 reflects a portion of the light to reference detector 76.
  • Beam splitters 73,75 divide the transmitted light into the three waveguides 60,62,64 which are coated with a sensing chemistry 12a,b,c, respectively.
  • This arrangement can be used in two approaches: (1) Three different analytes can be analyzed simultaneously or (2) If there is a question of specificity, then coincidence or redundant analyses can be accomplished using two or more enzymes which respond to the target analyte.
  • the number of waveguides that can be used is only restricted by source strength and geometric considerations.
  • HMs heavy metals
  • Phosphatase, glucose oxidase, pyruvate oxidase, alcohol dehydrogenase and lactate dehydrogenase are other enzymes inhibited by HMs. This propensity can be used to create many more versatile sensors.
  • Urease is an enzyme which hydrolyzes urea according to the following reactions:
  • Reaction (1) is predominant at pHs above 8, reaction (2) at pHs between 6 and 7, and reaction (3) at pHs below 7. In order to monitor the rate of the reaction, the consumption of hydrogen ion has been measured.
  • the membranes when in contact with ammonia, assume the blue color of the BTB anion and this can be monitored in absorption photometrically at 605 nm.
  • the time for the reaction to go to 95% completion (t 95 ) is » 100 minutes. Fortunately, it is not necessary to wait for completion and good results can be obtained by calibrating the system at a fixed time after the initiation of the reaction or by doing kinetic slope measurements.
  • bromothymol blue is replaced by a fluorescent dye of a pKa similar to that of BTB (7.2), a fluorescent 18 sensor is obtained.
  • a fluorescent dye of similar pKa is l-hydroxypyrene-3,6,8-trisulfonate with a pKa of 7.3. It can be excited at 460 nm and fluoresces above 500 nm with a maximum at 512 nm.
  • the cuvette with « 2 ⁇ m pvc coating on one of the inner walls was exposed to air for 15 minutes for further drying.
  • the cuvettes were activated in 1 mM hydrochloric acid for 10 minutes and another 10 minutes in a 0.1 M solution of ammonium chloride.
  • a fluorescent sensor for ammonium ion is described in U.S. Patent Application Serial No. 08/009,171 which is herein incorporated by reference.
  • Figure 7 is the response curve of the ammonia sensor at various NH 3 concentration as a function of time and absorbance when measured at 605 nm.
  • Urease is strongly inhibited by Hg 2+ , Ag + and Cu 2+ and also inactivated by other heavy metal ions.
  • the inhibition of urease by Ag + is shown in Figure 10.
  • the relative responses of the urease inhibition to several key heavy metals are shown in Figures 11a through llj with the solid black dots. This data is obtained when the reactants are immobilized without covalent bonds.
  • the open dots represent the behavious of the system with covalent immobilization.
  • the covalently bonded material behaves very poorly when compared to simple immobilization. Since the behavior of the covalently bonded materials is predictable, as a function of analyte concentration, this can be used as an internal reference.
  • Organometals e.g. mercury-organic compounds inhibit enzymes.

Abstract

Un détecteur optique destiné à détecter une première espèce comprend un détecteur optique qui produit un signal optique variant en fonction de la concentration d'une seconde espèce, un substrat chimique, et une enzyme qui agit sur le substrat chimique pour produire la seconde espèce et qui est inhibée par la première espèce de façon qu'elle ne puisse pas agir sur le substrat pour produire la seconde espèce. En outre, un procédé permettant de détecter une première espèce consiste à exposer un substrat chimique à un agent qui agit sur le substrat pour produire une seconde espèce et qui est inhibé par la première espèce de sorte qu'il n'agisse par sur le substrat pour produire la seconde espèce, à détecter optiquement les variations de la concentration de la seconde espèce, et à déterminer les variations de la première espèce à partir des variations détectées de la seconde espèce.
PCT/US1995/000846 1994-01-21 1995-01-20 Detecteurs proptochimiques dont le fonctionnement est fonde sur l'inhibition enzymatique WO1995020051A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US18487494A 1994-01-21 1994-01-21
US184,874 1994-01-21

Publications (1)

Publication Number Publication Date
WO1995020051A1 true WO1995020051A1 (fr) 1995-07-27

Family

ID=22678714

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1995/000846 WO1995020051A1 (fr) 1994-01-21 1995-01-20 Detecteurs proptochimiques dont le fonctionnement est fonde sur l'inhibition enzymatique

Country Status (1)

Country Link
WO (1) WO1995020051A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10164429A1 (de) * 2001-12-29 2003-07-17 It Dr Gambert Gmbh Vorrichtung zur Messung enzymatischer Bestandteile im Körperinneren von Mensch oder Tier
US6694067B1 (en) 2001-01-05 2004-02-17 Los Gatos Research Cavity enhanced fiber optic and waveguide chemical sensor
CN112525893A (zh) * 2019-09-19 2021-03-19 首都师范大学 一种基于纳米金催化能力的比色法传感器检测肝脏中铜离子的方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990004016A1 (fr) * 1988-10-14 1990-04-19 Univ Paris Vii Optrodes a enzymes immobilises
EP0538053A1 (fr) * 1991-10-18 1993-04-21 Gec-Marconi Limited Séparation et analyse
WO1993011259A1 (fr) * 1991-12-02 1993-06-10 Oriental Yeast Co., Ltd. Procede et reactif pour determiner le fer serique ou la capacite de fixation de fer sans saturation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990004016A1 (fr) * 1988-10-14 1990-04-19 Univ Paris Vii Optrodes a enzymes immobilises
EP0538053A1 (fr) * 1991-10-18 1993-04-21 Gec-Marconi Limited Séparation et analyse
WO1993011259A1 (fr) * 1991-12-02 1993-06-10 Oriental Yeast Co., Ltd. Procede et reactif pour determiner le fer serique ou la capacite de fixation de fer sans saturation

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
M. A. ARNOLD: "Fiber optic biosensing probes for biomedically important compounds", PROCEEDINGS OF SPIE - THE INTERNATIONAL SOCIETY FOR OPTICAL ENGINEERING, OPTICAL FIBERS IN MEDICINE III, vol. 906, LOS ANGELES, US, pages 128 - 133 *
O. S. WOLFBEIS & H. LI: "Fluorescence optical urea biosensor with an ammonium optrode as transducer", BIOSENSORS BIOELECTRONICS, vol. 8, no. 3-4, pages 161 - 166 *
O. S. WOLFBEIS: "optrodes for measuring enzyme activity and inhibition", NATO ASI SERIALS, SER. E, USES OF IMMOBILIZED BIOLOGICAL COMPOUNDS, vol. 252, pages 335 - 344 *
W. TRETTNAK ET AL: "Fiber-optic remote detection of pesticides and related inhibitors of the enzyme acetylcholine esterase", SENSORS AND ACTUATORS B, vol. b11, no. 1-3, pages 87 - 93 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6694067B1 (en) 2001-01-05 2004-02-17 Los Gatos Research Cavity enhanced fiber optic and waveguide chemical sensor
DE10164429A1 (de) * 2001-12-29 2003-07-17 It Dr Gambert Gmbh Vorrichtung zur Messung enzymatischer Bestandteile im Körperinneren von Mensch oder Tier
DE10164429B4 (de) * 2001-12-29 2006-09-07 It Dr. Gambert Gmbh Vorrichtung zur Messung enzymatischer Bestandteile im Körperinneren von Mensch oder Tier
CN112525893A (zh) * 2019-09-19 2021-03-19 首都师范大学 一种基于纳米金催化能力的比色法传感器检测肝脏中铜离子的方法

Similar Documents

Publication Publication Date Title
Choi Progress in enzyme-based biosensors using optical transducers
Monk et al. Optical fiber-based biosensors
D'Orazio Biosensors in clinical chemistry
Vadgama et al. Biosensors: recent trends. A review
Trettnak et al. Fibre-optic glucose sensor with a pH optrode as the transducer
Silvestre et al. Enzyme based assays in a sequential injection format: A review
US20060121547A1 (en) Diffusion layer for an enzyme-based sensor application
Walters et al. Fiber-optic biosensor for ethanol, based on an internal enzyme concept
Marazuela et al. Free cholesterol fiber-optic biosensor for serum samples with simplex optimization
Wolfbeis et al. Fluorescence optical urea biosensor with an ammonium optrode as transducer
Guilbault [41] Enzyme electrodes and solid surface fluorescence methods
AU670099B2 (en) Sensor for optical assay
US10024797B2 (en) Biosensing systems for measurement of lactose
Schaffar et al. Chemically mediated fiberoptic biosensors
Arnold Fluorophore-and chromophore-based fiberoptic biosensors
Kivirand et al. Calibrating biosensors in flow-through set-ups: studies with glucose optrodes
Abdel-Latif et al. Fiber optic sensors: recent developments
WO1995020051A1 (fr) Detecteurs proptochimiques dont le fonctionnement est fonde sur l'inhibition enzymatique
Taniai et al. Fluorometric determination of ethanol in liquor samples by flow-injection analysis using an immobilized enzyme-reactor column with packing prepared by coupling alcohol oxidase and peroxidase onto chitosan beads
US6649361B1 (en) Surface plasmon resonance enzyme sensor
Nagl et al. Classification of chemical sensors and biosensors based on fluorescence and phosphorescence
Schubert et al. Fibre optic fluorometric enzyme sensors for hydrogen peroxide and lactate, based on horseradish peroxidase and lactate oxidase
EP1027602B1 (fr) Procede pour determiner la concentration d'un analyte a l'aide d'un bioelement et d'un transducteur, et un dispositif a volume petit utilise dans le procede
Newman et al. Historical perspective of biosensor and biochip development
Abdel-Latif et al. Fiber optic-based biosensors utilizing immobilized enzyme systems

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA JP KR

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase