CA1244085A - Enzyme electrode type sensor for analyte determination - Google Patents
Enzyme electrode type sensor for analyte determinationInfo
- Publication number
- CA1244085A CA1244085A CA000518277A CA518277A CA1244085A CA 1244085 A CA1244085 A CA 1244085A CA 000518277 A CA000518277 A CA 000518277A CA 518277 A CA518277 A CA 518277A CA 1244085 A CA1244085 A CA 1244085A
- Authority
- CA
- Canada
- Prior art keywords
- layer
- enzyme
- membrane
- sensor
- analyte
- 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.)
- Expired
Links
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
- C12Q1/001—Enzyme electrodes
- C12Q1/002—Electrode membranes
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/817—Enzyme or microbe electrode
Abstract
Abstract Sensor A sensor of the enzyme-electrode type containing a layer of porous material of restricted permeability between the enzyme and a sample to be analysed. The porous material has a percentage porosity not greater than 5% and preferably in the range 0.005% to 0.5%.
Description
l B 33625 Sensor This invention relates to a sensor of the enzyme electrode type comprising an improved membrane and to an analytical method using the sensor.
Enzyme electrodes are increasingly used in medical and other laboratories particularly for the determination of materials such as glucose and urea in specimens of blood and other physiological fluids. Such electrodes are described in many publications notably an article by Clark and Lyons (Annals 10 of the New York Academy of Science, 102, 29-45, 1962) and US
Patents 3539455 and 3979274 to Clark and Newman respectively.
Enzyme electrodes are generally used to determine materials which themselves are not electrochemically active but which in the presence of suitable enzymes take part in reactions which produce species which can be readily detected by the electrodes. In enzyme electrodes the enzymes are rrequently located within polymeric materials in close proximity to the underlying electrode.
A considerable amount of research has been carried out in order to improve ~he properties of membranes for use in enzyme electrodes and many membranes for this purpose have been disclosed. An example of a type of membrane whlch is often used is the laminated membrane disclosed by Newman in US Patent 3979274. This membrane comprises a first or inner layer of an essentially homogeneous material, for example cellulose acetate, which can prevent the passage of materials of low molecular weight likely to interfere with the enzymic signal, a close adherent layer of the enzyme itself (with or without such other materials that may be blended with it), and a second layer (in this instance an outer layer) of a porous support film which can prevent the passage of cellular and colloidal elements.
The determination of glucose can be taken as an example of the determination of a material by an enzyme electrode. In the presence of the en~yme glucose oxidase the ~Z~ 35
Enzyme electrodes are increasingly used in medical and other laboratories particularly for the determination of materials such as glucose and urea in specimens of blood and other physiological fluids. Such electrodes are described in many publications notably an article by Clark and Lyons (Annals 10 of the New York Academy of Science, 102, 29-45, 1962) and US
Patents 3539455 and 3979274 to Clark and Newman respectively.
Enzyme electrodes are generally used to determine materials which themselves are not electrochemically active but which in the presence of suitable enzymes take part in reactions which produce species which can be readily detected by the electrodes. In enzyme electrodes the enzymes are rrequently located within polymeric materials in close proximity to the underlying electrode.
A considerable amount of research has been carried out in order to improve ~he properties of membranes for use in enzyme electrodes and many membranes for this purpose have been disclosed. An example of a type of membrane whlch is often used is the laminated membrane disclosed by Newman in US Patent 3979274. This membrane comprises a first or inner layer of an essentially homogeneous material, for example cellulose acetate, which can prevent the passage of materials of low molecular weight likely to interfere with the enzymic signal, a close adherent layer of the enzyme itself (with or without such other materials that may be blended with it), and a second layer (in this instance an outer layer) of a porous support film which can prevent the passage of cellular and colloidal elements.
The determination of glucose can be taken as an example of the determination of a material by an enzyme electrode. In the presence of the en~yme glucose oxidase the ~Z~ 35
2 B 33625 following reactlon occurs:-Glucos~ ~o glucose Gluconic a~ld +H 0oxidase The hydrogen peroxide produced in this reaction passes through the firs~ layer of a membrane such as that of ~S Patent 3979274 and can be determined using the electrode. Since the hydrogen peroxide produced is dependent upon the glucose present in a specimen, the glucose concentration can be determined using a suitably calibrated sensor.
To date a number of difficulties have limited the utility of enzyme electrodes and restricted the scale of their use in routine analysis of, e.g. blood samples.
Significant among these difficulties is the limited linearity of the response of electrodes to analytes such as glucose or lactate which are substrates for the enzyme catalysed reactions. The response is linear only over a limited range of low concentrations o the anal.ytes and hence the concentratlons of the materials to be determined must be low and generally diluted samples must be used in specimens for analysis using enzyme electrodes. It is not always prac~icable to make diluted samples for routine analysis outside the laboratory and it would be impossible for invasive monitoring.
According to the present invention we provide a sensor of the enzyme - electrode type for the determination of an analyte, said analyte being convertable in the presence of an enzyme into a species which can be detected by the sensor, which comprises an electrode and a membrane permeable to liquids and solutes positioned between the electrode and a specimen containing the analyte, said membrane comprising a layer containing one or more enzymes and a layer of material positioned between the enzyme - containing layer and the specimen characterised in that said layer of material contains an area through which analyte can pass formed from a porous material of restricted permeability having a porosity which is
To date a number of difficulties have limited the utility of enzyme electrodes and restricted the scale of their use in routine analysis of, e.g. blood samples.
Significant among these difficulties is the limited linearity of the response of electrodes to analytes such as glucose or lactate which are substrates for the enzyme catalysed reactions. The response is linear only over a limited range of low concentrations o the anal.ytes and hence the concentratlons of the materials to be determined must be low and generally diluted samples must be used in specimens for analysis using enzyme electrodes. It is not always prac~icable to make diluted samples for routine analysis outside the laboratory and it would be impossible for invasive monitoring.
According to the present invention we provide a sensor of the enzyme - electrode type for the determination of an analyte, said analyte being convertable in the presence of an enzyme into a species which can be detected by the sensor, which comprises an electrode and a membrane permeable to liquids and solutes positioned between the electrode and a specimen containing the analyte, said membrane comprising a layer containing one or more enzymes and a layer of material positioned between the enzyme - containing layer and the specimen characterised in that said layer of material contains an area through which analyte can pass formed from a porous material of restricted permeability having a porosity which is
3 B 33625 not greater than 5%.
Further according to the invention we provide a method for determining an analyte in a specimen when comprises contacting the specimen with the outer layer of a membrane, permeable to liquids and solutes and comprising one or more enzymes 9 in ~he presence of which the analyte is convertable into a species detectable by a sensor which incorporates the membrane, and one or more layers of material, and measuring the response of the sensor to the species, characterised in that a layer in the membrane between the enzyme and the specimen contains an area through which analyte can pass formed from a porous material of restricted permeability having a porosity which is not greater than 5%.
The area formed from a porous material having a porosity which is not greater than 5% causes the layer containing it to have restricted permeability. Preferably all or a ma~or proportion of the effective area of this layer is formed from msterial having a poroslty which is not greater than 5%.
In its most simple form the membrane in the sensor of the invention consists of the enzyme - containing layer and the layer oE resCricted permesbility. The layer of restricted permeability is the outer layer in this simple form of membrane and is contacted directly by the specimen in the method of the invention for dëtermining an analyte.
However it is preEerred that the membrane is a laminated membra~e of the type of which that disclosed in US
Patent 3979274 is an example. Such a membrane comprises a first or inner layer of material positioned between the enzyme-containing layer and the electrode, the enzyme-containing layer and a second layer of material on the other sid~ of the enzyme-containing layer which second layer is the layer having restricted permeability.
Hereafter in this specification the sensor of the invention which is described will contain a laminated membrane 8~
Further according to the invention we provide a method for determining an analyte in a specimen when comprises contacting the specimen with the outer layer of a membrane, permeable to liquids and solutes and comprising one or more enzymes 9 in ~he presence of which the analyte is convertable into a species detectable by a sensor which incorporates the membrane, and one or more layers of material, and measuring the response of the sensor to the species, characterised in that a layer in the membrane between the enzyme and the specimen contains an area through which analyte can pass formed from a porous material of restricted permeability having a porosity which is not greater than 5%.
The area formed from a porous material having a porosity which is not greater than 5% causes the layer containing it to have restricted permeability. Preferably all or a ma~or proportion of the effective area of this layer is formed from msterial having a poroslty which is not greater than 5%.
In its most simple form the membrane in the sensor of the invention consists of the enzyme - containing layer and the layer oE resCricted permesbility. The layer of restricted permeability is the outer layer in this simple form of membrane and is contacted directly by the specimen in the method of the invention for dëtermining an analyte.
However it is preEerred that the membrane is a laminated membra~e of the type of which that disclosed in US
Patent 3979274 is an example. Such a membrane comprises a first or inner layer of material positioned between the enzyme-containing layer and the electrode, the enzyme-containing layer and a second layer of material on the other sid~ of the enzyme-containing layer which second layer is the layer having restricted permeability.
Hereafter in this specification the sensor of the invention which is described will contain a laminated membrane 8~
4 B 33625 of the type of which the membrane described in US Patent 3979274 is an example having first and second layers the layer comprising the porous material having restricted permeability being the second layer.
It should be understood that the membranes in the sensor of the invention can contain more than two layers of material in addition to the enzyme-containing layer. For instance the second layer, i.e. of restricted per~eability, is not necessarily the outermost layer of the membrane. There may be a further layer or layers of material, i.e. third, fourth etc layers, between the second layer or layer of restricted permeability and the specimen. Often however the second layer will be the outer layer and its outer face will be contacted by the specimen.
Generally the porous material of restricted permeability~uysed in the second layer will be a polymeric material but other suitable materials may be used. Thus the second layer may be formed from glass or a me~al having pores cut by lasers.
Suitably the second layer of material is formed from material having a porosity not greater than 2~. Very low porosities are preferred for instance in the range 0.001% or 0-005~ to 0.5~, Percentage porosity is the product of pore area X
pore density X100. Most suitable porous materials of low percentage porosity will have pores of mean diameter less than 0.03 microns, preferably 0.01 to 0.03 microns. However mater~als having pores of mean diameter greater than 0.03 microns can be used successfully if the pore density is reduced. Thus materials having mean pore diameters ~p to 0.2 microns can be used. Examples of rated pore size and rated pore densities for materials of suitable low porosities are as follows:-:~2~
Rated Pore ¦ Rated Pore ¦ Calculated ¦ Size (microns) ¦ Density (pores/cm2) I Porosity 1 O.l 1 3 x 108 1 2.3 0-05 1 6 x 108 1 1.1 0-03 1 6 2 lO~ I 0.42 0.01 1 6 x 1O8 1 0-047 1 0.01 1 1 x 1~8 1 ~.008 Other material parameters which may be manipulated to produce material of suitably restricted permeability for the second layer of porous material include pore tortuosity. The thickness of the second layer also influences permeability.
In the sensor of the invention the second layer of the membrane acts as a diffusion barrier and prevents or restricts the passage of compounds of high molecular weight and gives strength ~o the membrane sufficient to enable it to retain its shape and to maintain suitable contact with the electrode.
Suitable porous materiAls ~o~r the second layer include porous polycarbonates, polyurethanes, and modified cellulose particularly cellulose nitrate, cellulose acetate and regenerated cellulose.
Suitable materials also lnclude materials having molecular weight cut offs of 20,000 or less. To ensure rapid electrode responses the thickness of the second layer is preferably less than 20 microns, especially in the range 1 to 10 microns. Especially suitable materials for the second layer ha~e pores of mean diameter within the range 0.015 microns to 0.025 microns.
The sensor of the invention may have a detachable membrane or it may be a disposable sensor with an adherent membrane. Naterials used in the formation of suitable electrodes for the sensors include inert metals and/or carbon.
When the sensor incorporates a laminated membrane of the type disclosed in US Patent 3979274 the first layer which is to be located between the enzyme layer and the electrode is suitably formed from polymethyl-methacrylate, polyurethane, cellulose acetate or another porous material which will restrict or prevent passage of electroactive interfering compounds such as ascorbic acid and tyrosine. Suitably the first layer has a thickness in the range 0.2 microns to 1.0 microns.
The enzyme present in the sensor of the invention may be located in the membrane in any suitable manner.
Preferably in a laminated membrane it is present between the first and second layers of porous material and forms the bond between them. In this situation, and also generally, the enzyme i9 preferably immobilised by mixing with a material which causes cross linking to occur. A very suitable material for thls purpose is glutaraldehyde; proteins such as albumin and other materials may also be included. In order to facilitate the obtaining of rapid stable readings from the sen~or it is preferred that the enzy~e-containing layer is thin, i.e. not greater than 5 microns thick.
The enzyme to be used in the sensor of the invention will depend upon the analyte whose concentration is to be determined. If the analyte is glucose then the enzyme will be for example glucose oxidase. Other enzymes which may be present include uricase and lactate oxidase for determination of uric acid and lactic acid respectively. Enzyme systems comprising two or more enzymes may also be present.
A laminated membrane for use in the sensor of the invention for the determination of glucose may be prepared by a method including the following steps:
1. 1 mg glucose oxidase i5 dissolved in 50 ~1 of (100 mg/ml) albumin:
2. 3 ~1 of 12.5% glutaraldehyde solution is mixed with 3 ~1 of the enzyme/albumin mixture on a glass microscope slide: 5 3. 1 ~1 of the mixture produced in the previous step is ~.2~ 5 7 B 33~25 applied to one face of a 1 cm2 polycarbonate film having a porosity ~ot greater than 5% and pores with a mean diameter belo,w 0.03 microns:
4. The other surface of the enzyme layer is covered i~mediately with a thin cellulose acetate film and the resulting S laminated membrane ~s clamped for 3 minutes between glass ~lides. ~fter remo~al from the glass slides the laminated membrane produced by the above sequence of steps may be applied to a platinum electrode to form the sensor of the invention, the cellulose acetate film being nearest to the electrode and forming the first layer.
Use of the method of the invention gives the advantage of an increase in the concentration range over whlch a graph of con-centration against sensor response is linear. With conventional methods linearity wa~ generally extended only up to approximately a concentration of 3 m mol per litre for glucose.
Using the method of the invention linearity is increased and the range extends to glucose concentrationY of 5~ m mol per litre and even higher. At the higher concentrations this is achieved through restriction oE s-lbstrate entry into the enzyme layer and therefore with some loss of sensitivity. Thus the range covers the concentrations of glucose which can be anticipated in blood samples thereby enabling blood glucose levels to be determined more readlly. Thls ls a considerable advantage in situatlons where large numbers of determination~ must be made reg~larly and with minimal sample preparation. Linearity is also extended by applying to the second layer of the membrane a medium comprising an organo-silane having reactive groups.
Such a treatment may be applied to the second layer of the me~brane in the sensor of the present invention ~o produce a combined effect and further improved linearity.
In the drawings:
Figure 1 shows the sensor of the invention in cross-section; and Figures 2-6 are graphs showing the relationship between current density and the glucose or lactate concentration in solutions.
In Figure 1, reference numeral 1 is ehe second layer of the membrane formed from a polycarbonate film having a porosity of 0.42%, 2 is a layer of glucose oxidase enzyme dis~olved in albumin and mixed with glutaraldehyde, 3 is the S first layer formed from cellulose acetate, 4 is the platinum working electrode and 5 is the silver reference electrode. 1, 2 and 3 together form a laminated membrane. Platinum working electrode 4 acts as an anode whilst silver reference electrode
It should be understood that the membranes in the sensor of the invention can contain more than two layers of material in addition to the enzyme-containing layer. For instance the second layer, i.e. of restricted per~eability, is not necessarily the outermost layer of the membrane. There may be a further layer or layers of material, i.e. third, fourth etc layers, between the second layer or layer of restricted permeability and the specimen. Often however the second layer will be the outer layer and its outer face will be contacted by the specimen.
Generally the porous material of restricted permeability~uysed in the second layer will be a polymeric material but other suitable materials may be used. Thus the second layer may be formed from glass or a me~al having pores cut by lasers.
Suitably the second layer of material is formed from material having a porosity not greater than 2~. Very low porosities are preferred for instance in the range 0.001% or 0-005~ to 0.5~, Percentage porosity is the product of pore area X
pore density X100. Most suitable porous materials of low percentage porosity will have pores of mean diameter less than 0.03 microns, preferably 0.01 to 0.03 microns. However mater~als having pores of mean diameter greater than 0.03 microns can be used successfully if the pore density is reduced. Thus materials having mean pore diameters ~p to 0.2 microns can be used. Examples of rated pore size and rated pore densities for materials of suitable low porosities are as follows:-:~2~
Rated Pore ¦ Rated Pore ¦ Calculated ¦ Size (microns) ¦ Density (pores/cm2) I Porosity 1 O.l 1 3 x 108 1 2.3 0-05 1 6 x 108 1 1.1 0-03 1 6 2 lO~ I 0.42 0.01 1 6 x 1O8 1 0-047 1 0.01 1 1 x 1~8 1 ~.008 Other material parameters which may be manipulated to produce material of suitably restricted permeability for the second layer of porous material include pore tortuosity. The thickness of the second layer also influences permeability.
In the sensor of the invention the second layer of the membrane acts as a diffusion barrier and prevents or restricts the passage of compounds of high molecular weight and gives strength ~o the membrane sufficient to enable it to retain its shape and to maintain suitable contact with the electrode.
Suitable porous materiAls ~o~r the second layer include porous polycarbonates, polyurethanes, and modified cellulose particularly cellulose nitrate, cellulose acetate and regenerated cellulose.
Suitable materials also lnclude materials having molecular weight cut offs of 20,000 or less. To ensure rapid electrode responses the thickness of the second layer is preferably less than 20 microns, especially in the range 1 to 10 microns. Especially suitable materials for the second layer ha~e pores of mean diameter within the range 0.015 microns to 0.025 microns.
The sensor of the invention may have a detachable membrane or it may be a disposable sensor with an adherent membrane. Naterials used in the formation of suitable electrodes for the sensors include inert metals and/or carbon.
When the sensor incorporates a laminated membrane of the type disclosed in US Patent 3979274 the first layer which is to be located between the enzyme layer and the electrode is suitably formed from polymethyl-methacrylate, polyurethane, cellulose acetate or another porous material which will restrict or prevent passage of electroactive interfering compounds such as ascorbic acid and tyrosine. Suitably the first layer has a thickness in the range 0.2 microns to 1.0 microns.
The enzyme present in the sensor of the invention may be located in the membrane in any suitable manner.
Preferably in a laminated membrane it is present between the first and second layers of porous material and forms the bond between them. In this situation, and also generally, the enzyme i9 preferably immobilised by mixing with a material which causes cross linking to occur. A very suitable material for thls purpose is glutaraldehyde; proteins such as albumin and other materials may also be included. In order to facilitate the obtaining of rapid stable readings from the sen~or it is preferred that the enzy~e-containing layer is thin, i.e. not greater than 5 microns thick.
The enzyme to be used in the sensor of the invention will depend upon the analyte whose concentration is to be determined. If the analyte is glucose then the enzyme will be for example glucose oxidase. Other enzymes which may be present include uricase and lactate oxidase for determination of uric acid and lactic acid respectively. Enzyme systems comprising two or more enzymes may also be present.
A laminated membrane for use in the sensor of the invention for the determination of glucose may be prepared by a method including the following steps:
1. 1 mg glucose oxidase i5 dissolved in 50 ~1 of (100 mg/ml) albumin:
2. 3 ~1 of 12.5% glutaraldehyde solution is mixed with 3 ~1 of the enzyme/albumin mixture on a glass microscope slide: 5 3. 1 ~1 of the mixture produced in the previous step is ~.2~ 5 7 B 33~25 applied to one face of a 1 cm2 polycarbonate film having a porosity ~ot greater than 5% and pores with a mean diameter belo,w 0.03 microns:
4. The other surface of the enzyme layer is covered i~mediately with a thin cellulose acetate film and the resulting S laminated membrane ~s clamped for 3 minutes between glass ~lides. ~fter remo~al from the glass slides the laminated membrane produced by the above sequence of steps may be applied to a platinum electrode to form the sensor of the invention, the cellulose acetate film being nearest to the electrode and forming the first layer.
Use of the method of the invention gives the advantage of an increase in the concentration range over whlch a graph of con-centration against sensor response is linear. With conventional methods linearity wa~ generally extended only up to approximately a concentration of 3 m mol per litre for glucose.
Using the method of the invention linearity is increased and the range extends to glucose concentrationY of 5~ m mol per litre and even higher. At the higher concentrations this is achieved through restriction oE s-lbstrate entry into the enzyme layer and therefore with some loss of sensitivity. Thus the range covers the concentrations of glucose which can be anticipated in blood samples thereby enabling blood glucose levels to be determined more readlly. Thls ls a considerable advantage in situatlons where large numbers of determination~ must be made reg~larly and with minimal sample preparation. Linearity is also extended by applying to the second layer of the membrane a medium comprising an organo-silane having reactive groups.
Such a treatment may be applied to the second layer of the me~brane in the sensor of the present invention ~o produce a combined effect and further improved linearity.
In the drawings:
Figure 1 shows the sensor of the invention in cross-section; and Figures 2-6 are graphs showing the relationship between current density and the glucose or lactate concentration in solutions.
In Figure 1, reference numeral 1 is ehe second layer of the membrane formed from a polycarbonate film having a porosity of 0.42%, 2 is a layer of glucose oxidase enzyme dis~olved in albumin and mixed with glutaraldehyde, 3 is the S first layer formed from cellulose acetate, 4 is the platinum working electrode and 5 is the silver reference electrode. 1, 2 and 3 together form a laminated membrane. Platinum working electrode 4 acts as an anode whilst silver reference electrode
5 acts as a cathode. The membrane is held in place on the electrode by a perspe~ ring pressing down on outer layer 1 towards its outer edges at 6.
The use of the sensor shown in Figure 1 is illustrated in the following examples:-Example 1 An aqueous solution containing 30 mg/ml glucose oxidase enzyme (E.C.1.1.3.4) and 200 mg/ml albumin was mixed with an equal volume of a solution containing 50 ~g/ml glutaraldehyde. A 1 cm2 plece of a "NUCLEPORE"*polycarbonate film t~ean pore diameter ~ 0.05 microns) was exposed to 5 ~l of the mixed solutions in order to impregnate the film with the en~yme. In this instance the film acts as a support for the enzyme and has no effect on the linearlty of the response obtained from the sensor.
The enzyme-impregnated film was placed over the working and reference electrodes of the sensor (which had previously been moistened with 0.067 M phosphate buffer containing 50 m mol per litre sodium chloride at p~ 7.4 to ensure electrolytic contact between the working and reference electrodes). The film to be studied as the second layer was placed over the enzyme impregnated film. The screw-fit top of the electrode body was then positioned and tightened putting sli~ht tension on the resulting laminated membrane. The membrane was then tested. Since the upper Eilm (the film to be be tested as the second layer) may be replacPd with other films, it was possible ~o use the same supported enzyme layer * Reg TM
~z~ s for several different second layers).
In this example the films tested as the second layer were as follows:-(1) A further 0.05 micron mean pore diameter "NUCLEPORE"
polycarbonate film:
(2) A 0.015 micron mean pore diamter "NUCLEPOR~"
polycarbonate membrane; and (3) A regenerated cellulose film made by Schleicher and Schull (RC52) having a mean pore diameter of 0.01 microns.
~ith film (1) the linearity of the sensor was 7 m mol per litre. The results with fi.lms (2) and (3~ are given in Figures 2 and 3 respecti~ely of the accompanying drawings which are graphs of current density in ~ A (ordinates) against glucose concentration in m mol per litre (abcisses). These show that, using films (2) and (3) as the second layer, the sensor shows much greater linearity - greater than 20 m mol per litre for (2) and at least 50 m mol per litre for (3).
Response time (98%) for (2) was 30-60 seconds and for (3) 20 minutes. Response time (90%) for (3) was 7 minutes.
Example 2 Example 1 was repeated with a lactate oxidase enzyme (E.C.1.1.3.2.). The films tested as the second layer were those described as (1) and (2) in Example 1, having mean pore dismeters of 0.05 microns and 0.015 microns respectively.
With film (1) the linearity of the sensor was 0.2 m mol per litre. The result with film (2) is shown in Figure 4 of the accompanying drawings which is a graph of current density in ~ A (ordinate) against glucose concentration in m mol per litre (abcissa). Again linearity using the smaller pore film as the second layer is much greater than when the larger pore film is used. In this instance a linearity of at least 4 m mol per litre is achieved with film (2), the response time being 1-2 minutes.
The method of Example 1 was used to test as the second layer a regenerated cellulose film made by SchleLcher and Schull (RC52) having a designated pore size in the range 0.005 to 0.010 ~um. The result is shown in Figure 5 ~ ich is a graph of current density in luA (ordinate) against glucose concentration in m mole per litre (abcissa). This shows that the response of the sensor is linear over a substantial concPntration range.
The method of Example 1 was used to test as the second layer a cellulose acetate film made by Schleicher and Schull (AC62) having a designated pore size in the range 0.005 to 0.01 ~m. The result is shown in Figure 6 which is a graph of curren~ density in ~ (ordinate) against glucose con-centration in m mol per litre (abcissa). This shows that the response is linear over a substantial concentration range.
The use of the sensor shown in Figure 1 is illustrated in the following examples:-Example 1 An aqueous solution containing 30 mg/ml glucose oxidase enzyme (E.C.1.1.3.4) and 200 mg/ml albumin was mixed with an equal volume of a solution containing 50 ~g/ml glutaraldehyde. A 1 cm2 plece of a "NUCLEPORE"*polycarbonate film t~ean pore diameter ~ 0.05 microns) was exposed to 5 ~l of the mixed solutions in order to impregnate the film with the en~yme. In this instance the film acts as a support for the enzyme and has no effect on the linearlty of the response obtained from the sensor.
The enzyme-impregnated film was placed over the working and reference electrodes of the sensor (which had previously been moistened with 0.067 M phosphate buffer containing 50 m mol per litre sodium chloride at p~ 7.4 to ensure electrolytic contact between the working and reference electrodes). The film to be studied as the second layer was placed over the enzyme impregnated film. The screw-fit top of the electrode body was then positioned and tightened putting sli~ht tension on the resulting laminated membrane. The membrane was then tested. Since the upper Eilm (the film to be be tested as the second layer) may be replacPd with other films, it was possible ~o use the same supported enzyme layer * Reg TM
~z~ s for several different second layers).
In this example the films tested as the second layer were as follows:-(1) A further 0.05 micron mean pore diameter "NUCLEPORE"
polycarbonate film:
(2) A 0.015 micron mean pore diamter "NUCLEPOR~"
polycarbonate membrane; and (3) A regenerated cellulose film made by Schleicher and Schull (RC52) having a mean pore diameter of 0.01 microns.
~ith film (1) the linearity of the sensor was 7 m mol per litre. The results with fi.lms (2) and (3~ are given in Figures 2 and 3 respecti~ely of the accompanying drawings which are graphs of current density in ~ A (ordinates) against glucose concentration in m mol per litre (abcisses). These show that, using films (2) and (3) as the second layer, the sensor shows much greater linearity - greater than 20 m mol per litre for (2) and at least 50 m mol per litre for (3).
Response time (98%) for (2) was 30-60 seconds and for (3) 20 minutes. Response time (90%) for (3) was 7 minutes.
Example 2 Example 1 was repeated with a lactate oxidase enzyme (E.C.1.1.3.2.). The films tested as the second layer were those described as (1) and (2) in Example 1, having mean pore dismeters of 0.05 microns and 0.015 microns respectively.
With film (1) the linearity of the sensor was 0.2 m mol per litre. The result with film (2) is shown in Figure 4 of the accompanying drawings which is a graph of current density in ~ A (ordinate) against glucose concentration in m mol per litre (abcissa). Again linearity using the smaller pore film as the second layer is much greater than when the larger pore film is used. In this instance a linearity of at least 4 m mol per litre is achieved with film (2), the response time being 1-2 minutes.
The method of Example 1 was used to test as the second layer a regenerated cellulose film made by SchleLcher and Schull (RC52) having a designated pore size in the range 0.005 to 0.010 ~um. The result is shown in Figure 5 ~ ich is a graph of current density in luA (ordinate) against glucose concentration in m mole per litre (abcissa). This shows that the response of the sensor is linear over a substantial concPntration range.
The method of Example 1 was used to test as the second layer a cellulose acetate film made by Schleicher and Schull (AC62) having a designated pore size in the range 0.005 to 0.01 ~m. The result is shown in Figure 6 which is a graph of curren~ density in ~ (ordinate) against glucose con-centration in m mol per litre (abcissa). This shows that the response is linear over a substantial concentration range.
Claims (10)
1. A sensor of the enzyme-electrode type for the determination of an analyte, said analyte being convertable in the presence of an enzyme into a species which can be detected by the sensor, which comprises an electrode and a membrane permeable to liquids and solutes positioned between the electrode and a specimen containing the analyte, said membrane comprising a layer containing one or more enzymes and a layer of material positioned between the enzyme-containing layer and the specimen wherein said layer of material contains an area through which analyte can pass formed from a porous material of restricted permeability having a porosity which is not greater than 5%.
2. A sensor according to claim 1 which incorporates a laminated membrane comprising an enzyme-containing layer situated between a first layer of material and a second layer of material, the first layer being between the enzyme-containing layer and an electrode in the sensor.
3. A sensor according to claim 2 wherein the first layer of the membrane is formed from a polymeric material selected from the group consisting of polymethyl methacrylate, cellulose acetate and polyurethane.
4. A sensor according to claim 1 wherein the porous material of restricted permeability is formed from a polymeric material selected from the group consisting of polycarbonates and a modified cellulose.
5. A sensor according to claim 1 wherein the porous material of restricted permeability forms the total effective area of the layer containing it.
6. A sensor according to claim 1 wherein the porous material of restricted permeability has pores of mean diameter less than 0.03 microns.
7. A sensor according to claim 6 wherein the pores have a mean diameter in the range 0.01 to 0.03 microns.
8. A sensor according to claim 1 wherein the porous material of restricted permeability has a porosity not greater than 2%.
9. A sensor according to claim 8 wherein the porosity is in the range 0.005% to 0.5%.
10. A method for determining an analyte in a specimen with the outer layer of a membrane, permeable to liquids and solutes and comprising one or more enzymes, in the presence of which the analyte is convertable into a species detectable by a sensor which incorporates the membrane, and one or more layers of material, and measuring the response of the sensor to the species, wherein a layer in the membrane between the enzyme and the specimen contains an area through which analyte can pass formed from a porous material of restricted permeability having a porosity which is not greater than 5%.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB858522834A GB8522834D0 (en) | 1985-09-16 | 1985-09-16 | Sensor |
GB8522834 | 1985-09-16 |
Publications (1)
Publication Number | Publication Date |
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CA1244085A true CA1244085A (en) | 1988-11-01 |
Family
ID=10585224
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA000518277A Expired CA1244085A (en) | 1985-09-16 | 1986-09-16 | Enzyme electrode type sensor for analyte determination |
Country Status (17)
Country | Link |
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US (1) | US5437973A (en) |
EP (1) | EP0216577B1 (en) |
JP (1) | JPS6267442A (en) |
AT (1) | ATE73842T1 (en) |
AU (1) | AU605111B2 (en) |
CA (1) | CA1244085A (en) |
DE (1) | DE3684394D1 (en) |
DK (1) | DK169882B1 (en) |
ES (1) | ES2002348A6 (en) |
FI (1) | FI91023C (en) |
GB (1) | GB8522834D0 (en) |
GR (1) | GR862368B (en) |
IE (1) | IE71918B1 (en) |
NO (1) | NO863686L (en) |
NZ (1) | NZ217598A (en) |
PT (1) | PT83378B (en) |
ZA (1) | ZA867038B (en) |
Families Citing this family (78)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA1247700A (en) * | 1985-09-20 | 1988-12-28 | The Regents Of The University Of California | Two-dimensional diffusion glucose substrate sensing electrode |
GB8606824D0 (en) * | 1986-03-19 | 1986-04-23 | Univ Strathclyde | Biochemical detector |
US5352348A (en) * | 1987-04-09 | 1994-10-04 | Nova Biomedical Corporation | Method of using enzyme electrode |
US4759828A (en) * | 1987-04-09 | 1988-07-26 | Nova Biomedical Corporation | Glucose electrode and method of determining glucose |
GB8718430D0 (en) * | 1987-08-04 | 1987-09-09 | Ici Plc | Sensor |
DE68922414T2 (en) * | 1988-02-05 | 1995-11-09 | Nova Biomedical Corp | Enzyme electrode. |
DK61488D0 (en) * | 1988-02-05 | 1988-02-05 | Novo Industri As | COURSE OF ACTION |
US6306594B1 (en) | 1988-11-14 | 2001-10-23 | I-Stat Corporation | Methods for microdispensing patterened layers |
US5200051A (en) * | 1988-11-14 | 1993-04-06 | I-Stat Corporation | Wholly microfabricated biosensors and process for the manufacture and use thereof |
JPH0339648A (en) * | 1989-07-06 | 1991-02-20 | Nok Corp | Glucose biosensor |
JP2655727B2 (en) * | 1989-08-09 | 1997-09-24 | 日機装株式会社 | Enzyme sensor |
DK170103B1 (en) * | 1990-08-31 | 1995-05-22 | Radiometer Medical As | Electrochemical biosensor and biosensor membrane |
DE4208186C2 (en) * | 1992-03-11 | 1996-02-22 | Bst Bio Sensor Tech Gmbh | Sandwich membrane for biosensors and their use |
GB9211402D0 (en) * | 1992-05-29 | 1992-07-15 | Univ Manchester | Sensor devices |
GB9215971D0 (en) * | 1992-07-28 | 1992-09-09 | Univ Manchester | Sensor devices |
GB9215973D0 (en) * | 1992-07-28 | 1992-09-09 | Univ Manchester | Sensor devices |
AT399511B (en) * | 1992-10-29 | 1995-05-26 | Jobst Gerhard Ing | SENSOR FOR DETECTING BIOLOGICALLY IMPLEMENTABLE SUBSTANCES |
US5766839A (en) * | 1994-06-17 | 1998-06-16 | Ysi Incorporated | Processes for preparing barrier layer films for use in enzyme electrodes and films made thereby |
US5520788A (en) * | 1995-01-17 | 1996-05-28 | The Yellow Springs Instrument Company, Inc. | Support layer for enzyme electrode laminated membranes |
US5755231A (en) * | 1995-05-17 | 1998-05-26 | Plus Bio, Inc. | Test strip including integral specimen flow retarding structure |
EP0857107B1 (en) * | 1995-08-01 | 2004-09-29 | Ysi Incorporated | Improved laminated membrane structure for polarographic measurement and methods of making said structures |
DE19621241C2 (en) * | 1996-05-25 | 2000-03-16 | Manfred Kessler | Membrane electrode for measuring the glucose concentration in liquids |
US6020052A (en) * | 1996-07-30 | 2000-02-01 | Ysi Incorporated | Laminated membrane structure for polarographic measurement and methods of making said structures |
US5804048A (en) * | 1996-08-15 | 1998-09-08 | Via Medical Corporation | Electrode assembly for assaying glucose |
ATE227844T1 (en) | 1997-02-06 | 2002-11-15 | Therasense Inc | SMALL VOLUME SENSOR FOR IN-VITRO DETERMINATION |
US6764581B1 (en) | 1997-09-05 | 2004-07-20 | Abbott Laboratories | Electrode with thin working layer |
US6030827A (en) * | 1998-01-23 | 2000-02-29 | I-Stat Corporation | Microfabricated aperture-based sensor |
US8688188B2 (en) | 1998-04-30 | 2014-04-01 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
US6175752B1 (en) | 1998-04-30 | 2001-01-16 | Therasense, Inc. | Analyte monitoring device and methods of use |
US8346337B2 (en) | 1998-04-30 | 2013-01-01 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
US8465425B2 (en) | 1998-04-30 | 2013-06-18 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
US8974386B2 (en) | 1998-04-30 | 2015-03-10 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
US6949816B2 (en) | 2003-04-21 | 2005-09-27 | Motorola, Inc. | Semiconductor component having first surface area for electrically coupling to a semiconductor chip and second surface area for electrically coupling to a substrate, and method of manufacturing same |
US8480580B2 (en) | 1998-04-30 | 2013-07-09 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
US9066695B2 (en) | 1998-04-30 | 2015-06-30 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
US6251260B1 (en) | 1998-08-24 | 2001-06-26 | Therasense, Inc. | Potentiometric sensors for analytic determination |
US6591125B1 (en) | 2000-06-27 | 2003-07-08 | Therasense, Inc. | Small volume in vitro analyte sensor with diffusible or non-leachable redox mediator |
US6338790B1 (en) | 1998-10-08 | 2002-01-15 | Therasense, Inc. | Small volume in vitro analyte sensor with diffusible or non-leachable redox mediator |
EP1192269A2 (en) | 1999-06-18 | 2002-04-03 | Therasense, Inc. | MASS TRANSPORT LIMITED i IN VIVO /i ANALYTE SENSOR |
US6616819B1 (en) | 1999-11-04 | 2003-09-09 | Therasense, Inc. | Small volume in vitro analyte sensor and methods |
US6560471B1 (en) | 2001-01-02 | 2003-05-06 | Therasense, Inc. | Analyte monitoring device and methods of use |
EP1397068A2 (en) | 2001-04-02 | 2004-03-17 | Therasense, Inc. | Blood glucose tracking apparatus and methods |
US6960466B2 (en) | 2001-05-31 | 2005-11-01 | Instrumentation Laboratory Company | Composite membrane containing a cross-linked enzyme matrix for a biosensor |
AU2003303597A1 (en) | 2002-12-31 | 2004-07-29 | Therasense, Inc. | Continuous glucose monitoring system and methods of use |
US7587287B2 (en) | 2003-04-04 | 2009-09-08 | Abbott Diabetes Care Inc. | Method and system for transferring analyte test data |
US8066639B2 (en) | 2003-06-10 | 2011-11-29 | Abbott Diabetes Care Inc. | Glucose measuring device for use in personal area network |
EP1718198A4 (en) | 2004-02-17 | 2008-06-04 | Therasense Inc | Method and system for providing data communication in continuous glucose monitoring and management system |
WO2006109311A2 (en) * | 2005-04-15 | 2006-10-19 | Ramot At Tel Aviv University Ltd. | Enzyme-channeling based electrochemical biosensors |
US8112240B2 (en) | 2005-04-29 | 2012-02-07 | Abbott Diabetes Care Inc. | Method and apparatus for providing leak detection in data monitoring and management systems |
US7766829B2 (en) | 2005-11-04 | 2010-08-03 | Abbott Diabetes Care Inc. | Method and system for providing basal profile modification in analyte monitoring and management systems |
US7885698B2 (en) | 2006-02-28 | 2011-02-08 | Abbott Diabetes Care Inc. | Method and system for providing continuous calibration of implantable analyte sensors |
US7620438B2 (en) | 2006-03-31 | 2009-11-17 | Abbott Diabetes Care Inc. | Method and system for powering an electronic device |
US8226891B2 (en) | 2006-03-31 | 2012-07-24 | Abbott Diabetes Care Inc. | Analyte monitoring devices and methods therefor |
US7920907B2 (en) | 2006-06-07 | 2011-04-05 | Abbott Diabetes Care Inc. | Analyte monitoring system and method |
US8930203B2 (en) | 2007-02-18 | 2015-01-06 | Abbott Diabetes Care Inc. | Multi-function analyte test device and methods therefor |
US8732188B2 (en) | 2007-02-18 | 2014-05-20 | Abbott Diabetes Care Inc. | Method and system for providing contextual based medication dosage determination |
US8123686B2 (en) | 2007-03-01 | 2012-02-28 | Abbott Diabetes Care Inc. | Method and apparatus for providing rolling data in communication systems |
US8665091B2 (en) | 2007-05-08 | 2014-03-04 | Abbott Diabetes Care Inc. | Method and device for determining elapsed sensor life |
US8456301B2 (en) | 2007-05-08 | 2013-06-04 | Abbott Diabetes Care Inc. | Analyte monitoring system and methods |
US8461985B2 (en) | 2007-05-08 | 2013-06-11 | Abbott Diabetes Care Inc. | Analyte monitoring system and methods |
US7928850B2 (en) | 2007-05-08 | 2011-04-19 | Abbott Diabetes Care Inc. | Analyte monitoring system and methods |
WO2009051901A2 (en) * | 2007-08-30 | 2009-04-23 | Pepex Biomedical, Llc | Electrochemical sensor and method for manufacturing |
US8702932B2 (en) | 2007-08-30 | 2014-04-22 | Pepex Biomedical, Inc. | Electrochemical sensor and method for manufacturing |
WO2010056876A2 (en) | 2008-11-14 | 2010-05-20 | Pepex Biomedical, Llc | Manufacturing electrochemical sensor module |
US8951377B2 (en) | 2008-11-14 | 2015-02-10 | Pepex Biomedical, Inc. | Manufacturing electrochemical sensor module |
US9445755B2 (en) | 2008-11-14 | 2016-09-20 | Pepex Biomedical, Llc | Electrochemical sensor module |
US8103456B2 (en) | 2009-01-29 | 2012-01-24 | Abbott Diabetes Care Inc. | Method and device for early signal attenuation detection using blood glucose measurements |
US20100213057A1 (en) | 2009-02-26 | 2010-08-26 | Benjamin Feldman | Self-Powered Analyte Sensor |
US9226701B2 (en) | 2009-04-28 | 2016-01-05 | Abbott Diabetes Care Inc. | Error detection in critical repeating data in a wireless sensor system |
WO2010138856A1 (en) | 2009-05-29 | 2010-12-02 | Abbott Diabetes Care Inc. | Medical device antenna systems having external antenna configurations |
WO2011026148A1 (en) | 2009-08-31 | 2011-03-03 | Abbott Diabetes Care Inc. | Analyte monitoring system and methods for managing power and noise |
WO2011026147A1 (en) | 2009-08-31 | 2011-03-03 | Abbott Diabetes Care Inc. | Analyte signal processing device and methods |
US9320461B2 (en) | 2009-09-29 | 2016-04-26 | Abbott Diabetes Care Inc. | Method and apparatus for providing notification function in analyte monitoring systems |
WO2012162151A2 (en) | 2011-05-20 | 2012-11-29 | Pepex Biomedical, Inc. | Manufacturing electrochemical sensor modules |
JP6443802B2 (en) | 2011-11-07 | 2018-12-26 | アボット ダイアベティス ケア インコーポレイテッドAbbott Diabetes Care Inc. | Analyte monitoring apparatus and method |
US9968306B2 (en) | 2012-09-17 | 2018-05-15 | Abbott Diabetes Care Inc. | Methods and apparatuses for providing adverse condition notification with enhanced wireless communication range in analyte monitoring systems |
US11224367B2 (en) | 2012-12-03 | 2022-01-18 | Pepex Biomedical, Inc. | Sensor module and method of using a sensor module |
CN107003264B (en) | 2014-06-04 | 2020-02-21 | 普佩克斯生物医药有限公司 | Electrochemical sensor and method of manufacturing an electrochemical sensor using advanced printing techniques |
Family Cites Families (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3539455A (en) * | 1965-10-08 | 1970-11-10 | Leland C Clark Jr | Membrane polarographic electrode system and method with electrochemical compensation |
US3723064A (en) * | 1971-07-26 | 1973-03-27 | L Liotta | Method and device for determining the concentration of a material in a liquid |
CA1054034A (en) * | 1975-06-20 | 1979-05-08 | Barbara J. Bruschi | Multilayer analytical element |
US3979274A (en) * | 1975-09-24 | 1976-09-07 | The Yellow Springs Instrument Company, Inc. | Membrane for enzyme electrodes |
US4176333A (en) * | 1978-06-12 | 1979-11-27 | Westinghouse Electric Corp. | Magnetic core for single phase electrical inductive apparatus |
US4240438A (en) * | 1978-10-02 | 1980-12-23 | Wisconsin Alumni Research Foundation | Method for monitoring blood glucose levels and elements |
US4401122A (en) * | 1979-08-02 | 1983-08-30 | Children's Hospital Medical Center | Cutaneous methods of measuring body substances |
JPS5627643A (en) * | 1979-08-14 | 1981-03-18 | Toshiba Corp | Electrochemical measuring device |
US4356074A (en) * | 1980-08-25 | 1982-10-26 | The Yellow Springs Instrument Company, Inc. | Substrate specific galactose oxidase enzyme electrodes |
JPS57116164A (en) * | 1981-01-12 | 1982-07-20 | Nissan Motor Co Ltd | Method of igniting internal combustion engine |
US4418148A (en) * | 1981-11-05 | 1983-11-29 | Miles Laboratories, Inc. | Multilayer enzyme electrode membrane |
JPS59164953A (en) * | 1983-03-10 | 1984-09-18 | Fuji Electric Corp Res & Dev Ltd | Immobilized enzyme film and manufacture thereof |
AU564494B2 (en) * | 1983-05-05 | 1987-08-13 | Medisense Inc. | Enzyme cascade energy coupling assay |
US4522786A (en) * | 1983-08-10 | 1985-06-11 | E. I. Du Pont De Nemours And Company | Multilayered test device for detecting analytes in liquid test samples |
JPS60185153A (en) * | 1984-03-02 | 1985-09-20 | Fuji Electric Corp Res & Dev Ltd | Immobilized enzyme membrane |
JPS60185155A (en) * | 1984-03-02 | 1985-09-20 | Fuji Electric Corp Res & Dev Ltd | Analysis of glucose |
JPS61145447A (en) * | 1984-12-19 | 1986-07-03 | Fuji Electric Co Ltd | Immobilized enzyme membrane |
CA1247700A (en) * | 1985-09-20 | 1988-12-28 | The Regents Of The University Of California | Two-dimensional diffusion glucose substrate sensing electrode |
GB8718430D0 (en) * | 1987-08-04 | 1987-09-09 | Ici Plc | Sensor |
-
1985
- 1985-09-16 GB GB858522834A patent/GB8522834D0/en active Pending
-
1986
- 1986-09-11 DE DE8686307011T patent/DE3684394D1/en not_active Expired - Fee Related
- 1986-09-11 AT AT86307011T patent/ATE73842T1/en not_active IP Right Cessation
- 1986-09-11 EP EP86307011A patent/EP0216577B1/en not_active Expired - Lifetime
- 1986-09-16 ZA ZA867038A patent/ZA867038B/en unknown
- 1986-09-16 NZ NZ217598A patent/NZ217598A/en unknown
- 1986-09-16 NO NO863686A patent/NO863686L/en unknown
- 1986-09-16 IE IE247186A patent/IE71918B1/en not_active IP Right Cessation
- 1986-09-16 CA CA000518277A patent/CA1244085A/en not_active Expired
- 1986-09-16 DK DK444086A patent/DK169882B1/en active
- 1986-09-16 AU AU62729/86A patent/AU605111B2/en not_active Ceased
- 1986-09-16 PT PT83378A patent/PT83378B/en not_active IP Right Cessation
- 1986-09-16 ES ES8601932A patent/ES2002348A6/en not_active Expired
- 1986-09-16 JP JP61217853A patent/JPS6267442A/en active Pending
- 1986-09-16 GR GR862368A patent/GR862368B/en unknown
- 1986-09-16 FI FI863738A patent/FI91023C/en not_active IP Right Cessation
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1993
- 1993-05-07 US US08/058,037 patent/US5437973A/en not_active Expired - Fee Related
Also Published As
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IE862471L (en) | 1987-03-16 |
PT83378B (en) | 1993-05-31 |
FI863738A0 (en) | 1986-09-16 |
NZ217598A (en) | 1989-10-27 |
DK444086A (en) | 1987-03-17 |
ES2002348A6 (en) | 1988-08-01 |
JPS6267442A (en) | 1987-03-27 |
NO863686L (en) | 1987-03-17 |
GR862368B (en) | 1986-12-23 |
ATE73842T1 (en) | 1992-04-15 |
FI91023C (en) | 1994-04-25 |
AU605111B2 (en) | 1991-01-10 |
DK169882B1 (en) | 1995-03-20 |
FI91023B (en) | 1994-01-14 |
ZA867038B (en) | 1987-05-27 |
EP0216577B1 (en) | 1992-03-18 |
EP0216577A3 (en) | 1988-07-20 |
AU6272986A (en) | 1987-03-19 |
DE3684394D1 (en) | 1992-04-23 |
GB8522834D0 (en) | 1985-10-23 |
NO863686D0 (en) | 1986-09-16 |
EP0216577A2 (en) | 1987-04-01 |
PT83378A (en) | 1987-05-06 |
FI863738A (en) | 1987-03-17 |
US5437973A (en) | 1995-08-01 |
IE71918B1 (en) | 1997-03-12 |
DK444086D0 (en) | 1986-09-16 |
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