WO2001040788A1 - Biocapteur - Google Patents
Biocapteur Download PDFInfo
- Publication number
- WO2001040788A1 WO2001040788A1 PCT/JP2000/008508 JP0008508W WO0140788A1 WO 2001040788 A1 WO2001040788 A1 WO 2001040788A1 JP 0008508 W JP0008508 W JP 0008508W WO 0140788 A1 WO0140788 A1 WO 0140788A1
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- WO
- WIPO (PCT)
- Prior art keywords
- biosensor
- cavity
- side wall
- facing
- surfactant
- Prior art date
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/001—Enzyme electrodes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
Definitions
- the present invention relates to a biosensor for analyzing a specific component in a liquid sample, and more particularly to a biosensor provided with a cavity for introducing a liquid sample by capillary action.
- a blood glucose level is measured by measuring a current value obtained by a reaction between glucose in blood and a reagent such as glucose oxidase carried in the sensor. There is something that asks.
- FIG. 4 is an exploded perspective view showing the above-described conventional biosensor for measuring a blood glucose level.
- a working electrode 1 and a counter electrode 2 serving as electrodes are formed by printing on an insulating substrate 5 such as polyethylene terephthalate, and a reagent layer containing glucose oxidase and an electron acceptor is formed on these electrodes.
- a surfactant layer 11 made of egg yolk lecithin or the like is formed on the reagent layer 10.
- a spacer 7 in which a portion on the layer 10 is elongated and cut away, and a cover 6 having an air escape hole 9 are bonded on the insulating substrate 5.
- blood is introduced from the suction port 8 into the cavity 12 by capillary action, and is guided to a position where the electrode and the reagent layer 10 are present.
- the current value generated by the reaction between the blood and the reagent on the electrode is read by connecting to an external measuring device (not shown) through leads 3 and 4, and the blood glucose level in the blood is determined based on the current value. is there.
- the surfactant layer 11 is provided so as to cover the reagent layer 10 and blood is easily introduced into the cavity 12, the blood is dissolved while the surfactant layer 11 is dissolved.
- the surfactant layer 11 is introduced into the reaction layer 12 and dissolves the reagent layer 10 and reacts on the electrode. Therefore, the surfactant layer 11 inhibits the dissolution of the reagent layer 10 in blood, and thereby the sensor This causes variations in the sensitivity and measured values of the sensor, which adversely affects the sensor performance.
- a solution containing a reagent and an electron acceptor is developed and dried on an electrode to form a reagent layer 10 and a surfactant layer 11 is further formed thereon.
- a step of applying and spreading a solution containing a surfactant so as to cover the reagent layer 10 and a step of drying the surfactant layer are required, the biosensor is manufactured. There was also a problem that the process was time-consuming and productivity was poor.
- the present invention has been made in order to solve the above-mentioned conventional problems, and assists the flow of blood into the cavity without forming a surfactant layer on the reagent layer, thereby quickly and sufficiently introducing the blood. It is an object of the present invention to provide a biosensor capable of performing the above. Disclosure of the invention
- the biosensor according to claim 1 of the present invention has a cavity in which a liquid sample is introduced by capillary action, and can analyze components in the liquid sample by a reaction between the introduced liquid sample and a reagent.
- a liquid sample is introduced by capillary action
- the side wall itself of the side wall of the sensor facing the cavity has hydrophilicity.
- the biosensor having such a configuration, at least a part of the side walls of the sensor facing the cavity into which the liquid sample is introduced by the capillary phenomenon has hydrophilicity. Aspiration of a liquid sample can be assisted without providing a surfactant layer on the reagent that reacts with the sample. In addition, the manufacturing process of the sensor can be simplified.
- a side wall of the sensor facing the cavity is formed of a resin material mixed with a surfactant.
- a biosensor according to claim 3 of the present invention is the biosensor according to claim 2, wherein the amount of the surfactant is 0.01% by weight or more.
- the side wall of the sensor facing the cavity is formed of a resin material containing a surfactant in an amount of 0.01% by weight or more. The effect can be obtained.
- the biosensor according to claim 4 of the present invention is the biosensor according to claim 1, wherein a side wall of the sensor facing the cavity has a surface coated with a surfactant. It is formed by.
- the side wall of the hydrophilic sensor is formed by the film whose surface is coated with the surfactant, so that the interface is formed on the reagent reacting with the liquid sample. It is possible to provide a biosensor capable of assisting aspiration of a liquid sample without providing an activator layer, and thereby simplifying the manufacturing process of the sensor.
- the biosensor according to claim 5 of the present invention is the biosensor according to claim 1, wherein the side wall of the sensor facing the cavity is a resin having a hydrophilic polar group and has a surface thereof. It is formed by a coated film.
- the side wall of the hydrophilic sensor is formed by the film whose surface is coated with the resin having the hydrophilic polar group, the reagent reacting with the liquid sample is formed. It is possible to provide a biosensor that can assist in aspiration of a liquid sample without providing a surfactant layer thereon, and can thereby simplify the manufacturing process of the sensor.
- the thickness of the resin having a polar group is set to several tens angstroms or more.
- the side wall of the sensor facing the cavity is formed by the film coated with the surfactant or the resin having the hydrophilic polar group.
- a good blood suction assisting effect can be obtained.
- the biosensor according to claim 7 of the present invention is the biosensor according to claim 1, wherein at least a part of the surface of the sidewall forming the cavity is chemically modified. Is what is being done.
- the biosensor having such a configuration, at least a part of the side wall forming the cavity is chemically modified to form the hydrophilic side wall of the sensor. Therefore, it is possible to provide a biosensor that can assist in aspiration of a liquid sample without providing a surfactant layer on a reagent that reacts with the liquid sample, and can thereby simplify the manufacturing process of the sensor. Can be.
- the biosensor according to claim 8 of the present invention is the biosensor according to claim 7, wherein any of plasma discharge treatment, coupling reaction treatment, ozone treatment, and ultraviolet treatment is performed.
- a hydrophilic functional group is formed on at least a part of the side wall facing the cavity.
- the biosensor having such a configuration, at least one of the side walls forming the cavity is subjected to plasma discharge treatment, coupling reaction treatment, ozone treatment, or ultraviolet treatment. Since a hydrophilic functional group is formed on the surface by performing a chemical surface treatment, at least a part of the side wall facing the cavity has a hydrophilic surface. Can be.
- the biosensor according to claim 9 of the present invention is the biosensor according to claim 1, wherein at least a part of the side wall of the side wall facing the cavity has a rough surface. Is what it is.
- the side wall of the cavity forming the cavity is formed. That is, since the surface of at least a part of the side wall is roughened to form the side wall of the hydrophilic sensor, the surface layer of the surfactant reacting with the liquid sample does not need to be provided. Accordingly, it is possible to provide a biosensor capable of assisting aspiration of a liquid sample and simplifying the manufacturing process of the sensor.
- the biosensor according to claim 10 of the present invention is the biosensor according to claim 9, wherein any one of sandblasting, electric discharge machining, non-glare treatment, matte treatment, and chemical plating is applied.
- a rough surface is formed on at least a part of the side wall facing the cavity.
- At least one of the side walls forming the cavity is subjected to sandblasting, electric discharge machining, non-glare treatment, matte treatment, or chemical plating. Since the rough surface is formed, at least a part of the side wall of the side wall facing the cavity can be made hydrophilic.
- the biosensor according to claim 11 of the present invention is the biosensor according to any one of claims 1 to 10, wherein a reagent that reacts with a liquid sample is formed.
- the surface of the substrate to be formed also has a hydrophilic property.
- the biosensor having such a configuration, not only the surface of at least a part of the side wall but also the surface of the substrate on which the reagent reacting with the liquid sample is formed have hydrophilicity. As a result, the area of the hydrophilic portion of the side wall facing the cavity is increased, and the liquid sample can be more efficiently introduced.
- the biosensor according to claim 12 of the present invention is the biosensor according to any one of claims 1 to 10, wherein a reaction between the liquid sample and the reagent is performed.
- the surface of the substrate on which the electrode to be detected is formed also has a hydrophilic property. According to the biosensor having such a configuration, not only the surface of at least a part of the side wall forming the cavity but also the substrate on which the electrode for detecting the reaction between the liquid sample and the reagent is formed. Since the surface is also made hydrophilic, the adhesion between the electrode and the substrate on which the electrode is formed is improved, the problem of electrode peeling is eliminated, and the reliability of the sensor is improved.
- the biosensor according to claim 13 of the present invention is the biosensor according to claim 12, wherein the surface of the substrate is formed with a rough surface, and the formed rough surface is formed. The surface level is set to 0.01 / im to 1 ⁇ m.
- the side wall of the side wall facing the sensor facing the cavity has a rough surface having irregularities of 0.001 m to 1 ⁇ m. Since it is formed, the adhesion can be improved.
- FIG. 1 is an exploded perspective view showing a biosensor for measuring a blood glucose level according to an embodiment of the present invention.
- FIG. 2 is a graph comparing the sensitivity of the sensor to blood in Example 1 of the present invention.
- FIG. 3 is a graph comparing the sensitivity of the sensor to blood in Example 2 of the present invention.
- FIG. 4 is an exploded perspective view showing a conventional blood glucose level measuring biosensor. BEST MODE FOR CARRYING OUT THE INVENTION
- Embodiment 1 of the present invention will be described with reference to FIG.
- FIG. 1 is an exploded perspective view of a biosensor according to Embodiment 1 of the present invention.
- the surfactant layer 11 formed on the reaction reagent layer 10 is eliminated.
- at least a portion of the side wall facing the cavity 12 into which blood is introduced, ie, the spacer 7 and the cover 6, facing the cavity 12, is itself hydrophilic. In order to support the introduction of blood.
- One of the methods is to form an insulating film material by kneading in advance a chemical substance having a surfactant activity such as a surfactant into a material such as polyethylene terephthalate ⁇ polycarbonate, and cover with the insulating film material. 6 and spacer 7.
- a chemical substance having a surfactant activity such as a surfactant into a material such as polyethylene terephthalate ⁇ polycarbonate
- the types of surfactants that can be expected to have the above-described effects after being kneaded into the insulating film material include carboxylates, sulfonates, carboxylates, and phosphate esters.
- Surfactants cationic amines such as primary amine salts, secondary amine salts, tertiary amine salts, and quaternary ammonium salts; amphoteric surfactants such as amino acid type and betaine type; and And non-ionic surfactants such as polyethylene glycol type and polyhydric alcohol type.
- the material of the cover 6 spacer 7 into which the surfactant can be mixed is, in addition to those described above, polybutylene terephthalate, polyamide, polychlorinated vinyl, polyvinylidene chloride, polyimide, and polyimide. And the like.
- the material for the cover 6 and the spacer 7 is kneaded with a chemical substance having a surfactant such as a surfactant, so that the cavity 12 into which blood is introduced is formed.
- the side wall facing that is, the portion facing the cavity 12 of the cover 6 and the spacer 7 is made hydrophilic, so that the wettability of the side wall of the cavity 12 is improved, and the blood collected from the suction port 8 is improved. Can be quickly and reliably introduced into the cavity 1 2.
- the surfactant layer 11 on the reagent layer 10 can be eliminated, and the manufacturing process of the biosensor can be simplified.
- Embodiment 2 will be described with reference to FIG.
- the surface active agent is kneaded into the material of the cover 6 spacer 7 so as to face the cavity 12 of the cover 6 spacer 7. 1 PTJ
- the portion is made hydrophilic
- the insulating film such as polyethylene terephthalate / polycarbonate which is the base material of the cover 6 and the spacer 7, the first embodiment is used.
- the insulating film is coated by applying the surfactant described in the above or laminating a resin having a hydrophilic hydrophilic group on the surface, and the portion of the cover 6 spacer 7 facing the cavity 12 is hydrophilic. It has a characteristic.
- examples of the resin having a hydrophilic polar group include acryl-based, polyester-based, and urethane-based resins.
- the material of the base material is not limited to the above-mentioned insulating film such as polyethylene terephthalate or polycarbonate.
- polybutylene terephthalate, polyamide, polyvinyl chloride, polyvinylidene chloride, polyimide, nylon and the like can also be used.
- a primer treatment with an organic titanium-based compound, a polyethyleneimine-based compound, an isocyanate-based compound, or the like is performed on an insulating film such as polyethylene terephthalate-polycarbonate or the like as a base material of the cover 6-spacer 7.
- an insulating film such as polyethylene terephthalate-polycarbonate or the like as a base material of the cover 6-spacer 7.
- a surfactant is applied on an insulating film serving as a base material of the cover 6 and the spacer 7, or a resin having a hydrophilic polar group on the surface is coated with a resin.
- the surfaces of the cover 6 and the spacer 7 are coated by lamination, and the side wall facing the cavity 12 into which blood is introduced, that is, the portion of the cover 6 and the spacer 7 facing the cavity 12 has hydrophilicity.
- the wettability of the side wall of the cavity 12 can be improved, and blood collected from the suction port 8 can be quickly and reliably introduced into the cavity 12.
- the surfactant layer 11 on the reagent layer 10 can be eliminated, and the manufacturing process of the biosensor can be simplified.
- the effect of assisting blood suction is determined by the thickness of the surfactant layer applied on the insulating film serving as the base material of the cover 6 and the spacer 7 or the resin layer having a hydrophilic hydrophilic polar group to be laminated. Is acceptable if the thickness of the slab is more than tens of angstroms To maintain the effect over a long period of time, it is desirable that the force be several hundred angstroms or more.
- Embodiment 3 will be described with reference to FIG.
- the surface of the cover 6 spacer 7 facing the cavity 12 is made hydrophilic by kneading the surfactant itself into the material of the cover 6 spacer 7.
- the surfaces of the cover 6 and the spacer 7 facing the cavity 12 are chemically surface-treated and processed to form the force bar 6 and the spacer 7.
- the portion facing the cavity 12 has hydrophilicity.
- corona discharge treatment or gross treatment which is a typical method of plasma discharge treatment
- a discharge treatment A hydrophilic functional group such as a carboxyl group, a hydroxyl group, or a carbonyl group is formed on the surface of the cover 6 facing the cavity 12 or the surface of the phosphor 7 to form the cover 6. It chemically modifies the material surface of sa7 to improve the surface wettability.
- polybutylene terephthalate polyamide, polyvinyl chloride, polyvinyl chloride, polyvinylidene chloride, polyimide , Nylon, etc. can be used.
- the surface of the cover 6 and the spacer 7 facing the cavity 12 into which blood is introduced is chemically modified by performing a chemical surface treatment and processing. Since the portions of the cover 6 and the spacer 7 facing the cavity 12 are made hydrophilic, the wettability of the side walls of the cavity 12 is improved, and the blood collected from the suction port 8 is improved. Can be quickly and reliably introduced into the cavity 12. Along with that, the surfactant layer 11 on the reagent layer 10 can be eliminated, and the manufacturing process of the biosensor can be simplified.
- Embodiment 4 will be described with reference to FIG.
- the surface of the cover 6 and the spacer 7 facing the cavity 12 is hydrophilic by kneading the surfactant itself into the material of the cover 6 and the spacer 7.
- the surface of the cover 6 spacer 7 facing the cavity 12 is roughened so that a fine and continuous rough surface (unevenness) is formed on the material surface.
- the cover 6 ⁇ spacer 7 has a hydrophilic portion at the portion facing the cavity 12.
- polyethylene terephthalate polycarbonate polybutylene terephthalate
- polyamide polyamide
- polyvinyl chloride polychloride, and the like
- Bilidene chloride, polyimide, nylon, etc. can be used.
- the cover 6 facing the cavity 12 is formed with a fine and continuous rough surface (irregularities) on the surface of the spacer 7, whereby the cover 6 is formed.
- the spacer facing the cavity 12 of the spacer 7 is made hydrophilic, so that the wettability of the side wall of the cavity 12 is improved, and the blood collected from the suction port 8 is quickly and reliably collected. Can be introduced inside. Along with this, the surfactant layer 11 on the reagent layer 10 can be eliminated, and the manufacturing process of the biosensor can be simplified.
- Embodiment 5 will be described with reference to FIG.
- the side walls of the cavities 12, that is, the covers 16 and the spacers 7 facing the cavities 12 are treated to have hydrophilicity.
- the cover 6 spacer 7 not only the cover 6 spacer 7 but also the surface of the insulating substrate 5 on which the working electrode 1, the counter electrode 2, and the reagent layer 10 are formed as described above have the hydrophilic property. The processing is performed.
- the insulating substrate 5 can be subjected to the hydrophilic treatment as described in the first to fourth embodiments to further assist in suction of the liquid sample.
- the adhesion between the insulating substrate 5 and the electrode can be dramatically improved. This has the effect of improving.
- cutout grooves for forming cavities 12 were formed on insulating substrate 5 on which a plurality of electrodes and reagent layers 10 were formed, at positions corresponding to the respective electrodes and reagents. If the sensor shown in Fig. 1 is obtained by bonding the spacer 7 and the cover 6 with the air escape hole 9 to it using a press or the like to obtain the sensor shown in Fig. 1, it will occur during the punching.
- the electrode had peeled off from the insulating substrate 5 or had a crack on the electrode due to the impact. This is because the electrodes were formed by printing a paste made of a conductive material on the insulating substrate 5 having a very small polarity.
- the insulating substrate 5 is also subjected to the hydrophilic treatment as described in the first to fourth embodiments, so that the surface of the material of the insulating substrate 5 originally having a very small polarity is given a polarity, and the electrode As material A paste made of a conductive material to be used is placed on the insulating substrate 5 to improve the adhesion, and it is possible to prevent the electrode from peeling off the insulating substrate 5 and to prevent the electrode from cracking.
- the insulating substrate 5 is subjected to the hydrophilic treatment.
- the suction of the blood collected from the suction port 8 can be further assisted as compared with the case where only the spacer 7 is subjected to the hydrophilic treatment.
- the insulating substrate 5 is subjected to a hydrophilic treatment to impart polarity to the insulating substrate 5, so that the adhesion of the electrodes to the insulating substrate 5 increases, and the This can prevent peeling from the insulating substrate 5 and cracks generated in the electrodes.
- the level of the rough surface (irregularity) at which the effect of adhesion can be expected is 0.001 ⁇ m or more. It is preferably in the range of 1 ⁇ m, particularly preferably from 0.11 / zm to 0.1 ⁇ .
- Electrode layer consisting of working electrode 1 and counter electrode 2 by screen printing on insulating substrate 5 made of polyethylene terephthalate that has been subjected to corona discharge treatment (electric power: 400 W, discharge treatment speed: 3 Om / min) Is formed thereon, and a reagent layer 10 containing an enzyme (glucose oxidase) and an electron carrier (potassium ferricyanide) is formed thereon. Then, a spacer 7 composed of polyethylene terephthalate and an anion surfactant are prepared in advance.
- Table 1 shows the blood aspiration capability of the sensor fabricated in this way.
- the suction port 8 used had a height of 0.15 mm and a width of 2.0 ram.
- the values in Table 1 indicate the time required for the blood to completely fill the groove, which is the capillary through which the blood is guided, in a severe environment (environmental temperature 5 ° (: hematocrit value 65%)). This indicates that the same blood suction assisting effect as that of the conventional sensor was obtained.
- Figure 2 compares the sensor sensitivities at blood glucose concentrations between 53 and 992 mg / dl.
- the sensitivity of the sensor of Example 1 was about 5% higher than that of the conventional sensor. This confirms that the dissolution of the surfactant layer 11 improved the solubility of the reagent layer 10 that reacts with blood.
- Table 2 compares the repetition accuracy (CV value) at the time of 10 measurements in FIG. From this result, it can be seen that the measurement variation (variation among the sensors) in the sensor of the first embodiment is greatly reduced compared to the measurement variation in the conventional sensor. , Table 2
- An electrode layer consisting of a working electrode 1 and a counter electrode 2 is provided by screen printing on an insulating substrate 5 made of polyethylene terephthalate, and an enzyme (glucose oxidase) and an electron carrier (ferricyanide) are provided thereon.
- an enzyme glucose oxidase
- an electron carrier ferrerricyanide
- a blood glucose level measuring sensor in which a groove was formed as a capillary tube through which blood was guided was produced by bonding with a cover 6 made of a film (surface wetting index: 54 dynZcm or more). A similar evaluation was performed.
- Table 3 compares the blood aspiration speeds of the sensors created as described above, and Fig. 3 compares the sensor sensitivities at blood glucose concentrations of 53 to 992 mg / dl. Yes, and Table 4 shows Table 3 compares the repeater sensor accuracy (CV value) after 10 measurements.
- the biosensor according to the present invention can be used as a biosensor for improving the sensitivity and variation of the sensor when analyzing a specific component in the liquid sample by introducing a liquid sample from the cavity of the sensor by capillary action. It is.
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
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US09/890,761 US6830669B2 (en) | 1999-12-03 | 2000-12-01 | Biosensor |
EP00979044A EP1156325B1 (en) | 1999-12-03 | 2000-12-01 | Biosensor |
DE60026352T DE60026352T2 (de) | 1999-12-03 | 2000-12-01 | Biosensor |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP11/344495 | 1999-12-03 | ||
JP34449599A JP2001159618A (ja) | 1999-12-03 | 1999-12-03 | バイオセンサ |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2001040788A1 true WO2001040788A1 (fr) | 2001-06-07 |
Family
ID=18369721
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2000/008508 WO2001040788A1 (fr) | 1999-12-03 | 2000-12-01 | Biocapteur |
Country Status (6)
Country | Link |
---|---|
US (1) | US6830669B2 (ja) |
EP (1) | EP1156325B1 (ja) |
JP (1) | JP2001159618A (ja) |
CN (1) | CN1243976C (ja) |
DE (1) | DE60026352T2 (ja) |
WO (1) | WO2001040788A1 (ja) |
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Also Published As
Publication number | Publication date |
---|---|
JP2001159618A (ja) | 2001-06-12 |
EP1156325A1 (en) | 2001-11-21 |
US20040016642A1 (en) | 2004-01-29 |
DE60026352D1 (en) | 2006-04-27 |
CN1339105A (zh) | 2002-03-06 |
US6830669B2 (en) | 2004-12-14 |
EP1156325A4 (en) | 2002-07-17 |
DE60026352T2 (de) | 2006-10-19 |
CN1243976C (zh) | 2006-03-01 |
EP1156325B1 (en) | 2006-03-01 |
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