CN101382515B - Ethanol oxidizing enzyme film using polycarbonate film as substrate and method for making same - Google Patents
Ethanol oxidizing enzyme film using polycarbonate film as substrate and method for making same Download PDFInfo
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- CN101382515B CN101382515B CN200810225243XA CN200810225243A CN101382515B CN 101382515 B CN101382515 B CN 101382515B CN 200810225243X A CN200810225243X A CN 200810225243XA CN 200810225243 A CN200810225243 A CN 200810225243A CN 101382515 B CN101382515 B CN 101382515B
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 57
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 57
- 229920006289 polycarbonate film Polymers 0.000 title claims abstract description 16
- 239000000758 substrate Substances 0.000 title claims abstract description 6
- 238000000034 method Methods 0.000 title claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title abstract description 85
- 230000001590 oxidative effect Effects 0.000 title description 25
- 239000012528 membrane Substances 0.000 claims abstract description 75
- 239000004417 polycarbonate Substances 0.000 claims abstract description 50
- 229920000515 polycarbonate Polymers 0.000 claims abstract description 50
- 108010025188 Alcohol oxidase Proteins 0.000 claims abstract description 39
- 238000002360 preparation method Methods 0.000 claims abstract description 15
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000008055 phosphate buffer solution Substances 0.000 claims abstract description 11
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 6
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 22
- 239000000243 solution Substances 0.000 claims description 16
- 229910052697 platinum Inorganic materials 0.000 claims description 11
- 239000011148 porous material Substances 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 6
- 108010093096 Immobilized Enzymes Proteins 0.000 claims description 4
- 229920000642 polymer Polymers 0.000 claims description 3
- ZNZYKNKBJPZETN-WELNAUFTSA-N Dialdehyde 11678 Chemical compound N1C2=CC=CC=C2C2=C1[C@H](C[C@H](/C(=C/O)C(=O)OC)[C@@H](C=C)C=O)NCC2 ZNZYKNKBJPZETN-WELNAUFTSA-N 0.000 claims 1
- 239000007864 aqueous solution Substances 0.000 claims 1
- 230000004044 response Effects 0.000 abstract description 45
- 230000035945 sensitivity Effects 0.000 abstract description 8
- 238000003860 storage Methods 0.000 abstract description 4
- 108090000854 Oxidoreductases Proteins 0.000 abstract 1
- 102000004316 Oxidoreductases Human genes 0.000 abstract 1
- 238000001035 drying Methods 0.000 abstract 1
- 230000003100 immobilizing effect Effects 0.000 abstract 1
- 239000011159 matrix material Substances 0.000 description 25
- 238000012360 testing method Methods 0.000 description 21
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 6
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 229910021607 Silver chloride Inorganic materials 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000002848 electrochemical method Methods 0.000 description 4
- 239000003292 glue Substances 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- BHTJEPVNHUUIPV-UHFFFAOYSA-N pentanedial;hydrate Chemical compound O.O=CCCCC=O BHTJEPVNHUUIPV-UHFFFAOYSA-N 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 4
- 238000001514 detection method Methods 0.000 description 3
- 229940012466 egg shell membrane Drugs 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 2
- 102000004533 Endonucleases Human genes 0.000 description 2
- 108010042407 Endonucleases Proteins 0.000 description 2
- 235000013405 beer Nutrition 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 238000003822 preparative gas chromatography Methods 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
一种以聚碳酸酯膜为基体的乙醇氧化酶膜及其制备方法，属于电化学生物传感器及其制备技术领域。该乙醇氧化酶膜由聚碳酸酯基体膜、乙醇氧化酶及固定酶的戊二醛组成。该乙醇氧化酶膜的制备方法是：将聚碳酸酯膜粘在O型橡胶圈上，晾干后将该聚碳酸酯膜浸入磷酸缓冲溶液中，取出用高纯氮气吹干后滴加乙醇氧化酶，再通过戊二醛蒸气交联固定乙醇氧化酶制成酶膜。本发明乙醇氧化酶膜具有较短的响应时间、较高的灵敏度、较宽的线性响应范围以及良好的操作稳定性与储存稳定性；此外本发明的乙醇氧化酶膜制备方法简单、成本较低。
The invention discloses an alcohol oxidase film with a polycarbonate film as a substrate and a preparation method thereof, belonging to the technical field of electrochemical biosensors and its preparation. The alcohol oxidase film is composed of a polycarbonate base film, alcohol oxidase and glutaraldehyde immobilizing the enzyme. The preparation method of the ethanol oxidase film is as follows: stick the polycarbonate film on the O-shaped rubber ring, immerse the polycarbonate film in the phosphate buffer solution after drying, take it out and dry it with high-purity nitrogen, and add ethanol dropwise to oxidize it. Enzyme, and then cross-linked and immobilized alcohol oxidase by glutaraldehyde vapor to make an enzyme film. The alcohol oxidase membrane of the present invention has shorter response time, higher sensitivity, wider linear response range, and good operation stability and storage stability; in addition, the preparation method of the alcohol oxidase membrane of the present invention is simple and the cost is low .
Description
Technical field
The invention belongs to electrochemica biological sensor and preparing technical field thereof, particularly relating to a kind of is the ethanol oxidizing enzyme film and preparation method thereof of matrix with the polycarbonate membrane.
Background technology
The content that detects ethanol accurately and rapidly has significant application value at aspects such as food inspection, fermentation control and clinical diagnosises.The analyzing detecting method of ethanol has multiple, and wherein the most frequently used have vapor-phase chromatography, high performance liquid chromatography and spectrophotometric method etc., but these methods need expensive instrument and equipment more, and it is also complicated to detect step simultaneously.People begin to detect ethanol with electrochemica biological sensor in recent years, and this method is simple, quick and selectivity is higher.
It with the alcohol oxidase detection that the electrochemica biological sensor of bioactivator preparation can be used for ethanol, its principle of work is: under the effect of alcohol oxidase, dissolved oxygen DO in the solution can oxidation ethanol generates acetaldehyde and hydrogen peroxide, and the reduction by detecting dissolved oxygen DO in the solution or the growing amount of hydrogen peroxide can be determined the content of ethanol in the solution to be measured.
Because alcohol oxidase in use activity is held time shortlyer, realizes on the base film itself and the separating of electrode so usually alcohol oxidase is fixed on, and can change enzyme membrane behind enzyme deactivation more conveniently, thereby reduction sensor cost.Therefore screening fixing base material that suits and the process for fixation that improves enzyme is the important content that ethanol oxidizing enzyme film is studied with the recycling rate of waterused that keeps the active of enzyme and raising enzyme, is used to the fixing film matrix material of alcohol oxidase at present and comprises animals and plants film, cellulose acetate and derivatives membrane, polycarbonate membrane etc.
At document (1) Biosensors and Bioelectronics, 2007, among the 23:121, people such as Guangming Wen are that crosslinking chemical is fixed on alcohol oxidase on the egg shell membrane with the shitosan, be fixed in again and made the electrochemica biological sensor that detects ethanol on the oxygen electrode, based on the reduction detection concentration of ethanol of oxygen concentration in the solution.Studies show that the oxidation of ethanol endonuclease capable is fixed on the egg shell membrane preferably, the range of linearity that prepared biology sensor detects concentration of alcohol is 60 μ M～0.08mM, detects and is limited to 30 μ M (signal to noise ratio (S/N ratio) S/N=3), and the response time is 1 minute.The anti-interference of this electrochemica biological sensor is stronger, and has good operation and storage stability, and is consistent with the resulting testing result of usefulness vapor-phase chromatography to test result of samples.But adopt egg shell membrane as base film, in large-scale production, have bigger difficulty.
At document (2) Analytica Chimica Acta, 2002, among the 469:217, Dilek
Deng the people at first the using plasma polymerization technique be that the polycarbonate membrane of 0.03 μ m is handled and made its surface have amino with ethylenediamine to the aperture, then alcohol oxidase is fixed on the polycarbonate membrane of handling.Studies show that the oxidation of ethanol endonuclease capable is fixed on the polycarbonate membrane matrix preferably, is 5.6nA/mM based on the sensitivity of the electrochemica biological sensor of this enzyme membrane preparation, can the ethanolic solution of the following concentration of 2mM be detected, and the response time is 50 seconds.The parameter basically identical that the testing result and the beer manufacturer of beer sample given.But the using plasma polymerization technique carries out pre-service to polycarbonate membrane makes the preparation of enzyme membrane too complicated, adopts the polycarbonate membrane of smaller aperture due to increase resistance to mass tranfer in addition, is unfavorable for the diffusion of reactant.
Summary of the invention
The object of the present invention is to provide a kind of is the ethanol oxidizing enzyme film and preparation method thereof of matrix with the polycarbonate membrane.Polycarbonate membrane has excellent mechanical intensity and chemical resistance, and its thickness and aperture can in very large range regulate, and the production technology maturation has the commercially available prod of multiple different size available.It is matrix material that the present invention selects the commercial polycarbonate film in suitable depth and aperture, adopts glutaraldehyde steam cross-linking method that alcohol oxidase is fixed in the polycarbonate membrane surface, makes ethanol oxidizing enzyme film, has kept the activity of alcohol oxidase effectively.Electrochemica biological sensor based on this enzyme membrane preparation can be used for the detection of ethanol, and has short response time, higher sensitivity, the range of linearity and the good operational stability and the storage stability etc. of broad.
Ethanol oxidizing enzyme film of the present invention is made up of the polymer substance glutaraldehyde of polycarbonate membrane matrix, alcohol oxidase and immobilized enzyme, wherein the massfraction of polycarbonate membrane is 49.96～96.3%, the massfraction of alcohol oxidase be 3.35～41.7% and the unit area enzyme membrane in the content of alcohol oxidase be 12.7～31.2U/cm2, the massfraction of the polymer substance glutaraldehyde of immobilized enzyme is 0.35～8.34%; Described polycarbonate membrane average pore size scope is 0.10～0.80 μ m, and thickness range is 5～30 μ m.
The present invention is that the preparation method of the ethanol oxidizing enzyme film of matrix is with the polycarbonate membrane:
(1) being averaged pore diameter range is 0.10～0.80 μ m, thickness is that the commercial polycarbonate film of 5～30 μ m is cut into the disk than the big 2～3mm of employed platinum electrode cross-sectional diameter, subsequently the polycarbonate membrane disk that cuts is bonded on the O type rubber ring, dries stand-by;
(2) to immerse pH value be to handle 6～12 hours in 6.0～9.0 the phosphate buffer solution to the polycarbonate membrane that will glue O type rubber ring, and taking-up dries up with high pure nitrogen then;
(3) be 12.7～31.2U/cm by alcohol oxidase
2Consumption be that the alcohol oxidase drips of solution of 0.8～1.1U/ μ L is coated on the polycarbonate membrane of handling with concentration, in 0～4 ℃ refrigerator dry 0.5～2 hour;
(4) enzyme membrane is suspended on fills in the closed container that massfraction is 40～60% glutaraldehyde water solution and under 0～4 ℃ condition crosslinked 6～18 hours;
(5) enzyme membrane being immersed the pH value is that in conjunction with unstable alcohol oxidase, promptly forming with the polycarbonate membrane is the ethanol oxidizing enzyme film of matrix with flush away in 6.0～9.0 the phosphate buffer, is kept in 0～4 ℃ the refrigerator standby.
Effect of the present invention can be found out as the electrochemica biological sensor of the alcohol oxidase film production of matrix with polycarbonate membrane from using the present invention.Ethanol oxidizing enzyme film of the present invention is enclosed within the platinum electrode top as working electrode with O type rubber ring, and the platinum electrode conduct is to electrode, and the Ag/AgCl electrode is formed the ampere-type electrochemica biological sensor as contrast electrode.It is 6.0～9.0 phosphate buffer solution that the three-electrode system of sensor is placed the pH value, with the CHI660B electrochemical workstation this biology sensor is carried out electrochemical Characterization.Adopt the response current (as shown in Figure 1) of i-t method testing sensor to ethanol, ethanol oxidizing enzyme film of the present invention is higher to the sensitivity of ethanol response, and the response time is short; As seen from Figure 2, ethanol oxidizing enzyme film of the present invention is 0.01～2.5mmol/L to the linear response range of ethanol, greater than the linear response range of ethanol in document (1) and the document (2); Ethanol oxidizing enzyme film of the present invention is carried out continuous 200 times test (as shown in Figure 3), its response signal is not fallen as follows,, the big minor swing of response signal is in 5% in 200 times the test, and this explanation the present invention is that the ethanol oxidizing enzyme film of matrix has good operational stability with the polycarbonate membrane.
The invention has the advantages that: aspect the matrix selection, the present invention optimizes polycarbonate film thickness, pore size, selecting the average pore size scope for use is 0.10～0.80 μ m, thickness range is the polycarbonate membrane of 5～30 μ m, guaranteeing to help the rapid diffusion of reactant under the prerequisite effectively fixing, reduce resistance to mass tranfer to alcohol oxidase, thereby reduce the response time of enzyme membrane, realize the quick response of sensor substrate; Aspect the preparation method, alcohol oxidase membrane preparation method of the present invention is simple and easy to do, operation is less, cost is lower; At aspect of performance, ethanol oxidizing enzyme film of the present invention has short response time, higher sensitivity, linear response range and the good operational stability and the storage stability of broad.
Description of drawings
Fig. 1. the present invention is the i-t response curve of the ethanol oxidizing enzyme film of matrix to the ethanolic solution of 1mmol/L with the polycarbonate membrane.Wherein, horizontal ordinate-time t (unit: second, s); Ordinate-response current i (unit: microampere, μ A).
Fig. 2. concentration of alcohol and the present invention are the relation curve of response current of the ethanol oxidizing enzyme film of matrix with the polycarbonate membrane.Wherein, the concentration of horizontal ordinate-ethanolic solution (unit: mM/liter, mmol/L); Ordinate-response current i (unit: microampere, μ A).
Fig. 3. the present invention is the ethanol oxidizing enzyme film operational stability test pattern of matrix with the polycarbonate membrane.Wherein, horizontal ordinate-testing time (unit: inferior) ordinate-relative response (unit: %).
Fig. 4. the present invention is 30 secondary response situations before the ethanol oxidizing enzyme film stability test of matrix with the polycarbonate membrane.Wherein, horizontal ordinate-testing time (unit: inferior), ordinate-relative response (unit: %).
Embodiment:
(1) be 0.45 μ m with average pore size, thickness is the disk that the polycarbonate membrane of 10 μ m is cut into diameter 10mm, subsequently the polycarbonate membrane that cuts is bonded on the O type rubber ring, dries stand-by;
(2) to immerse pH value be to handle 6 hours in 7.5 the phosphate buffer solution to the polycarbonate membrane that will glue O type rubber ring, and taking-up dries up with high pure nitrogen then;
(3) be that the alcohol oxidase drips of solution of 1.02U/ μ L is coated on the polycarbonate membrane of handling with 5 μ L concentration, forming diameter was the enzyme layer circle spot of 5mm, with this enzyme membrane in 4 ℃ refrigerator dry 1 hour;
(4) enzyme membrane is suspended on fills in the closed container that massfraction is 50% glutaraldehyde water solution, and in 4 ℃ refrigerator crosslinked 12 hours;
(5) enzyme membrane being immersed the pH value is that in conjunction with unstable alcohol oxidase, promptly forming with the polycarbonate membrane is the ethanol oxidizing enzyme film of matrix with flush away in 7.5 the phosphate buffer, is kept in 4 ℃ the refrigerator standby.
The ethanol oxidizing enzyme film that will be matrix with the polycarbonate membrane is enclosed within the platinum electrode top as working electrode, the platinum electrode conduct is to electrode, the Ag/AgCl electrode is as contrast electrode, test system is the phosphate buffer solution of pH value 7.5, adopts the CHI660B electrochemical workstation to being that the electrochemica biological sensor of matrix carries out electrochemical Characterization with the polycarbonate membrane.Adopt the response current (as shown in Figure 1) of i-t method testing sensor to ethanol water, drawing its response sensitivity is 4.68 μ AL/mmol, 5 seconds response times; Sensor is 0.01～2.5mmol/L (as shown in Figure 2) to the range of linearity of ethanol water response; The signal response that follow-on test is 200 times as shown in Figure 3, Fig. 4 be preceding 30 times the test the signal response situation, as can be seen from the figure, follow-on test 200 secondary response signals are not seen obvious decline, the big minor swing of response signal is in 5% in 200 times test; This enzyme membrane follow-on test after 3 months its response signal still keep more than 80% of its initial response signal.
(1) be 0.10 μ m with average pore size, thickness is the disk that the polycarbonate membrane of 30 μ m is cut into diameter 10mm, subsequently the polycarbonate membrane that cuts is bonded on the O type rubber ring, dries stand-by;
(2) to immerse pH value be to handle 12 hours in 6.0 the phosphate buffer solution to the polycarbonate membrane that will glue O type rubber ring, and taking-up dries up with high pure nitrogen then;
(3) be that the alcohol oxidase drips of solution of 1.1U/ μ L is coated on the polycarbonate membrane of handling with 8 μ L concentration, forming diameter was the enzyme layer circle spot of 6mm, with this enzyme membrane in 0 ℃ refrigerator dry 2 hours;
(4) enzyme membrane is suspended on fills in the closed container that massfraction is 60% glutaraldehyde water solution, and in 0 ℃ refrigerator crosslinked 18 hours;
(5) enzyme membrane being immersed the pH value is that in conjunction with unstable alcohol oxidase, promptly forming with the polycarbonate membrane is the ethanol oxidizing enzyme film of matrix with flush away in 6.0 the phosphate buffer, is kept in 0 ℃ the refrigerator standby.
The ethanol oxidizing enzyme film that will be matrix with the polycarbonate membrane is enclosed within the platinum electrode top as working electrode, the platinum electrode conduct is to electrode, the Ag/AgCl electrode is as contrast electrode, test system is the phosphate buffer solution of pH value 6.0, adopts the CHI660B electrochemical workstation to being that the electrochemica biological sensor of matrix carries out electrochemical Characterization with the polycarbonate membrane.Adopt the response current of i-t method testing sensor to ethanol water, drawing its response sensitivity is 2.84 μ AL/mmol, 8 seconds response times; Sensor is 0.01～2.5mmol/L to the range of linearity of ethanol water response; This enzyme membrane follow-on test after 3 months its response signal still keep more than 70% of its initial response signal.
(1) be 0.80 μ m with average pore size, thickness is the disk that the polycarbonate membrane of 20 μ m is cut into diameter 10mm, subsequently the polycarbonate membrane that cuts is bonded on the O type rubber ring, dries stand-by;
(2) to immerse pH value be to handle 10 hours in 9.0 the phosphate buffer solution to the polycarbonate membrane that will glue O type rubber ring, and taking-up dries up with high pure nitrogen then;
(3) be that the alcohol oxidase drips of solution of 0.8U/ μ L is coated on the polycarbonate membrane of handling with 2 μ L concentration, forming diameter was the enzyme layer circle spot of 4mm, with this enzyme membrane in 0 ℃ refrigerator dry 0.5 hour;
(4) enzyme membrane is suspended on fills in the closed container that massfraction is 40% glutaraldehyde water solution, and in 2 ℃ refrigerator crosslinked 6 hours;
(5) enzyme membrane being immersed the pH value is that in conjunction with unstable alcohol oxidase, promptly forming with the polycarbonate membrane is the ethanol oxidizing enzyme film of matrix with flush away in 9.0 the phosphate buffer, is kept in 2 ℃ the refrigerator standby.
The ethanol oxidizing enzyme film that will be matrix with the polycarbonate membrane is enclosed within the platinum electrode top as working electrode, the platinum electrode conduct is to electrode, the Ag/AgCl electrode is as contrast electrode, test system is the phosphate buffer solution of pH value 9.0, adopts the CHI660B electrochemical workstation to being that the electrochemica biological sensor of matrix carries out electrochemical Characterization with the polycarbonate membrane.Adopt the response current of i-t method testing sensor to ethanol water, drawing its response sensitivity is 1.17 μ AL/mmol, and the response time is 10 seconds; Sensor is 0.01～2.5mmol/L to the range of linearity of ethanol water response; This enzyme membrane follow-on test after 3 months its response signal still keep more than 56% of its initial response signal.
Claims (3)
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| CN108226260A (en) * | 2017-12-27 | 2018-06-29 | 潘彦伶 | A kind of heavy metal quick-check sensor of cosmetics |
| US20210371845A1 (en) * | 2020-05-28 | 2021-12-02 | Nestor Maceda-Johnson | Synthesis Of Enzyme-Polymer Conjugates, Having Enhanced Activity & Stability |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59164953A (en) | 1983-03-10 | 1984-09-18 | Fuji Electric Corp Res & Dev Ltd | Immobilized enzyme membrane and its manufacturing method |
| JPH0359453A (en) | 1989-07-27 | 1991-03-14 | Kanzaki Paper Mfg Co Ltd | enzyme electrode |
| US5231028A (en) * | 1987-10-19 | 1993-07-27 | Cambridge Life Sciences Plc | Immobilized enzyme electrodes |
| WO2002002796A2 (en) * | 2000-07-05 | 2002-01-10 | Inventus Biotec Gesellschaft Für Innovative Bioanalytik, Biosensoren Und Diagnostika Mbh & Co. Kg | Disposable electrochemical bio-sensor for the quantitative determination of analyte concentrations in fluids |
| CA2402653A1 (en) * | 2002-09-26 | 2004-03-26 | Jane Dormon | Antiseptic coating for the prevention of disease transmission via biofilms |
| CN1563969A (en) * | 2004-03-22 | 2005-01-12 | 南开大学 | Biological enzyme electrode for biosensor, and its prepn. method |
| CN2723998Y (en) * | 2004-08-23 | 2005-09-07 | 南开大学 | Enzyme electrode for residual pesticide test |
-
2008
- 2008-10-29 CN CN200810225243XA patent/CN101382515B/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59164953A (en) | 1983-03-10 | 1984-09-18 | Fuji Electric Corp Res & Dev Ltd | Immobilized enzyme membrane and its manufacturing method |
| US5231028A (en) * | 1987-10-19 | 1993-07-27 | Cambridge Life Sciences Plc | Immobilized enzyme electrodes |
| JPH0359453A (en) | 1989-07-27 | 1991-03-14 | Kanzaki Paper Mfg Co Ltd | enzyme electrode |
| WO2002002796A2 (en) * | 2000-07-05 | 2002-01-10 | Inventus Biotec Gesellschaft Für Innovative Bioanalytik, Biosensoren Und Diagnostika Mbh & Co. Kg | Disposable electrochemical bio-sensor for the quantitative determination of analyte concentrations in fluids |
| CA2402653A1 (en) * | 2002-09-26 | 2004-03-26 | Jane Dormon | Antiseptic coating for the prevention of disease transmission via biofilms |
| CN1563969A (en) * | 2004-03-22 | 2005-01-12 | 南开大学 | Biological enzyme electrode for biosensor, and its prepn. method |
| CN2723998Y (en) * | 2004-08-23 | 2005-09-07 | 南开大学 | Enzyme electrode for residual pesticide test |
Non-Patent Citations (3)
| Title |
|---|
| Dilek Çökeliler等.Performance of amperometric alcohol electrodes.《Analytica Chimica Acta》.2002,第469卷(第2期),217-223. * |
| 周慧等.乙醇氧化酶电极的研究.《分析化学》.1993,第21卷(第12期),1471. * |
| 范培昌等.检测唾液中乙醇的玻态酶试条的研究.《分析科学学报》.2006,第22卷(第4期),431-434. * |
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