【発明の詳細な説明】 〔産業上の利用分野〕 本発明は、グルコースセンサに関する。更に詳しく
は、アンペロメトリ型のグルコースセンサに関する。Description: TECHNICAL FIELD The present invention relates to a glucose sensor. More particularly, it relates to an amperometric glucose sensor.
従来のアンペロメトリ型グルコースセンサにおいて
は、２個の電極を必要としており、その内の１個は対極
であり、他の１個は作用極であって、作用極にはグルコ
ースオキシダーゼ酵素（GOD）が固定化されている。こ
のセンサの原理は、グルコースとGODとの反応に伴ない
作用極上で発生する電極検知物質を定量することによ
り、グルコース濃度を決定し得る点にある。A conventional amperometric glucose sensor requires two electrodes, one of which is a counter electrode and the other one is a working electrode, and a glucose oxidase enzyme (GOD) is present at the working electrode. It is fixed. The principle of this sensor is that the glucose concentration can be determined by quantifying the electrode detection substance generated on the working electrode accompanying the reaction between glucose and GOD.
ところで、グルコースセンサの一つの形態として、針
状構造のものが挙げられるが、従来のアンペロメトリ型
グルコースセンサでは、これら２個の電極が例えばガラ
ス板などの平板状のものを平行に並べて構成させたもの
が代表的であり、生体内に挿入し易く血糖値などの測定
に好適に使用し得る針状電極としたものは未だ実用化さ
れていない。これは、形状そのものに基因する製作の困
難さや酵素固定化の困難さに由来しているものと考えら
れる。By the way, as one form of the glucose sensor, one having a needle-like structure can be mentioned. In the conventional amperometry type glucose sensor, these two electrodes are formed by arranging flat plates such as glass plates in parallel. A typical example is a needle-shaped electrode that is easily inserted into a living body and can be suitably used for measuring a blood glucose level and the like, and has not yet been put into practical use. It is considered that this is due to the difficulty of manufacturing and the immobilization of the enzyme due to the shape itself.
本発明の目的は、このような問題点を克服し、酵素固
定化や製作の容易性が得られるアンペロメトリ型グリコ
ースセンサを提供することにある。An object of the present invention is to provide an amperometric type glucose sensor that overcomes such problems and can be easily immobilized on an enzyme or manufactured.
かかる目的を達成せしめる本発明のグルコースセンサ
は、長さ方向に半截された中空針状対極の切断面に絶縁
層を介してグルコースオキシダーゼ酵素固定化平板状作
用極を酵素固定化面側を内側に向けて接着してなる。Glucose sensor of the present invention to achieve such an object, the glucose oxidase enzyme-immobilized plate-shaped working electrode through the insulating layer on the cut surface of the hollow needle-shaped counter electrode half-divided in the longitudinal direction, the enzyme-immobilized surface side inward. It is glued towards.
図面の第１図には、本発明に係るグルコースセンサの
一態様が接着前の状態で斜視図として示されている。即
ち、針状対極１は、一般に外径が約1mm以下の白金、
金、チタンなどの中空針の長さ方向の半截物２であっ
て、その先端部３が挿入し易いように注射針状に斜切さ
れている。そして、半截された切断面には、一般に接着
剤層を兼ねた絶縁層4,4′、例えばエポキシ樹脂接着
剤、シリコーン系接着剤あるいはガラスなどが塗布され
ている。In FIG. 1 of the drawings, one embodiment of the glucose sensor according to the present invention is shown as a perspective view before adhesion. That is, the needle-shaped counter electrode 1 is generally made of platinum having an outer diameter of about 1 mm or less,
A hollow needle 2 made of gold, titanium, or the like in the longitudinal direction, and a distal end portion 3 thereof is obliquely cut into an injection needle shape so as to be easily inserted. Insulating layers 4 and 4'which also double as an adhesive layer, such as an epoxy resin adhesive, a silicone adhesive, or glass, are applied to the cut surface.
一方、この絶縁層を介して中空針状対極に接着される
グルコースオキシダーゼ酵素固定化平板状作用極５は、
厚さ約0.05〜1mm程度の白金、チタンなどの平板６であ
って、一般にその長さが対極切断面の長さ以下で、幅が
切断面間隔に等しいものの片面側にGODを固定化７せし
めている。On the other hand, the glucose-oxidase enzyme-immobilized flat plate-shaped working electrode 5 adhered to the hollow needle-shaped counter electrode via this insulating layer is
A flat plate 6 made of platinum, titanium or the like having a thickness of about 0.05 to 1 mm, generally having a length equal to or less than the length of the opposite electrode cutting surface and having a width equal to the distance between the cutting surfaces. ing.
GODの作用極上への固定化に際しては、膜形成材料、
例えば酢酸セルロースなどのセルロース類、ポリビニル
ブチラール、ポリスルホンなどの合成高分子物質、ある
いはセルロース、キチン、アルブミン、アルギン酸ナト
リウム、アガロース、カラギーナンなどの天然高分子物
質が用いられ、それらを可溶性溶媒に溶解させたドープ
液として調製した後、浸漬法、ドロップ法、スピナ法な
どにより作用極面上に適用する。また、膜形成材料とし
て、分子中に光架橋性基としてスチルバゾリウム基、ジ
アソ基などの感光性基を有するポリビニルアルコールな
どの水溶性光架橋性重合体なども水溶液として用いるこ
とができる。When immobilizing GOD on the working electrode, a film-forming material,
For example, celluloses such as cellulose acetate, synthetic polymeric substances such as polyvinyl butyral and polysulfone, or natural polymeric substances such as cellulose, chitin, albumin, sodium alginate, agarose and carrageenan are used and dissolved in a soluble solvent. After prepared as a dope solution, it is applied on the working electrode surface by a dipping method, a drop method, a spinner method, or the like. As the film-forming material, a water-soluble photocrosslinkable polymer such as polyvinyl alcohol having a photosensitive group such as a stilbazolium group or a diazo group in the molecule as a photocrosslinkable group can also be used as an aqueous solution.
これらの膜形成材料によって形成される膜上へのGOD
の固定化は、グルタルアルデヒド、カルボジイミドなど
を用いる共有結合法、イオン結合法、吸着法、架橋法な
ど一般的に用いられている方法によって行われる。ある
いは、膜形成材料の溶液中などにGODを混合しておき、
膜形成時にこれらの固定化方法により固定化させること
もできる。この場合、光架橋性重合体が用いられたとき
には、それの光架橋はGODを失活させない波長である約3
50〜450nmの近紫外線による光照射によって行われる。GOD on the film formed by these film forming materials
The immobilization is carried out by a commonly used method such as a covalent bond method using glutaraldehyde, carbodiimide, etc., an ionic bond method, an adsorption method, a cross-linking method and the like. Alternatively, GOD is mixed in a solution of the film forming material,
It is also possible to immobilize by these immobilization methods at the time of film formation. In this case, when a photocrosslinkable polymer is used, its photocrosslinking is at a wavelength of about 3 which does not deactivate GOD.
It is performed by light irradiation with near ultraviolet rays of 50 to 450 nm.
このような固定化GODによるグルコースの定量は、なる反応により、生成したH2O2を作用極での酸化電流値
変化として検出する。Glucose quantification by such immobilized GOD, The H2 O2 generated by the reaction is detected as a change in the oxidation current value at the working electrode.
また、電子メディエータ（電子伝達体）であるフェロ
セン化合物、具体的にはフェロセン［ビス（シクロペン
タジエニル）鉄（II）］またはその誘導体、例えば1,
1′−ジメチルフェロセンなどを用い、それを作用極面
上に真空蒸着させた後、この蒸着面にGODを一般的に用
いられる固定化法によって固定化せしめ、あるいはGOD
を固定化させた後フェロセン化合物をそこに滴下法など
で重層し、この場合には、グルコース１分子が酸化されるのに、２
電子の作用極への移動が行われるので、これを電流値変
化として検出することができる。Further, a ferrocene compound which is an electron mediator (electron carrier), specifically, ferrocene [bis (cyclopentadienyl) iron (II)] or a derivative thereof, for example, 1,
Using 1'-dimethylferrocene, etc., vacuum-deposit it on the working electrode surface, and then fix the GOD on this deposition surface by a commonly used immobilization method.
After immobilizing, the ferrocene compound was overlaid there by a dropping method or the like, In this case, even if one glucose molecule is oxidized,
Since electrons are moved to the working electrode, this can be detected as a change in current value.
かかるGOD固定化平板状作用極の中空針状対極への接
着は、酵素固定化面側が内側に向けられるようにして行
われる。なお、対極および作用極からは、それぞれリー
ド線8,8′が引き出されており、測定時には、これらの
電極とは別に参照極も用いられる。The GOD-immobilized flat plate-shaped working electrode is adhered to the hollow needle-shaped counter electrode so that the enzyme-immobilized surface side faces inward. In addition, lead wires 8 and 8'are drawn out from the counter electrode and the working electrode, respectively, and a reference electrode is used in addition to these electrodes during measurement.
本発明に係る針状のアンペロメトリ型グルコースセン
サは、グルコースオキシダーゼ酵素の固定化が平板状の
作用極面上に対して行われるのでそこに困難性はなく、
また針状電極という形状に基因する製作上の困難性も格
別認められず、生体内に挿入し易く血糖値などの測定に
好適に使用し得るグルコースセンサとして提供される。The needle-shaped amperometry-type glucose sensor according to the present invention is not difficult because immobilization of the glucose oxidase enzyme is performed on the flat working electrode surface.
Further, no difficulty in manufacture due to the shape of the needle electrode is recognized, and the glucose sensor is provided as a glucose sensor that can be easily inserted into a living body and can be suitably used for measuring a blood glucose level and the like.
次に、実施例について本発明を説明する。 Next, the present invention will be described with reference to examples.
実施例 第１図に示された態様のグルコースセンサを製作し
た。中空針状対極は、内径0.8mm、外径1.0mm、長さ3cm
の白金中空針を半截し、先端部を斜切したものであり、
平板状作用極としては、1.0×2.5×0.2mmの寸法のもの
が用いられ、その片面側にGODを固定化させた。Example A glucose sensor of the embodiment shown in FIG. 1 was manufactured. The hollow needle-shaped counter electrode has an inner diameter of 0.8 mm, an outer diameter of 1.0 mm, and a length of 3 cm.
The platinum hollow needle of is cut in half and the tip is cut diagonally.
The flat working electrode having a size of 1.0 × 2.5 × 0.2 mm was used, and the GOD was immobilized on one side of the working electrode.
GODの固定化は、GOD30mgおよび牛血清アルブミン30mg
をpH7.0の5mMトリス−マレイン酸緩衝液0.4mlに溶解
し、その４μを平板状作用極面上に滴下し、４℃で４
時間放置した後、そこに1,1′−ジメチルフェロセンの
アセトン溶液４μを滴下し、再び４℃で４時間放置す
ることにより行われた。Immobilization of GOD is 30 mg GOD and 30 mg bovine serum albumin
Was dissolved in 0.4 ml of 5 mM Tris-maleic acid buffer having a pH of 7.0, and 4 μ of the solution was dropped on the flat working electrode surface, and the mixture was mixed at 4 ° C. for 4 hours.
After standing for 4 hours, 4 μm of an acetone solution of 1,1′-dimethylferrocene was added dropwise thereto, and the mixture was allowed to stand at 4 ° C. for 4 hours again.
このGOD固定化平板状作用極を、酵素固定化面側を内
側に向けて、中空針状対極にその切断面に塗布されたエ
ポキシ樹脂接着剤層を介して接着し、４℃で48時間放置
した。The GOD-immobilized flat plate-shaped working electrode was bonded to the hollow needle-shaped counter electrode with the epoxy resin adhesive layer applied to the cut surface, with the enzyme-immobilized surface side facing inward, and left at 4 ° C for 48 hours. did.
これらの各電極をポテンショスタットに接続し、対極
−作用極間電位50mV、作用極−参照極（Ag/AgCl電極）
間電位200mVとして、グルコースに対する応答を測定し
た。測定は、37℃でpH7.0の5mMトリス−マレイン酸緩衝
液を使用して行われた。その結果は、第２図のグラフに
示されるように、グルコース濃度50〜300mg/dlの範囲内
で、応答電流の定常値との間に直線的な相関関係が認め
られた。Each of these electrodes is connected to a potentiostat, the counter electrode-working electrode potential 50 mV, working electrode-reference electrode (Ag / AgCl electrode)
The response to glucose was measured with an inter-potential of 200 mV. The measurements were performed at 37 ° C. using 5 mM Tris-maleic acid buffer pH 7.0. As a result, as shown in the graph of FIG. 2, a linear correlation was observed with the steady-state value of the response current within the glucose concentration range of 50 to 300 mg / dl.
【図面の簡単な説明】 第１図は、本発明のグルコースセンサの一態様を接着前
の状態で示した斜視図である。また、第２図は、このグ
ルコースセンサを用いた場合のグルコース濃度と出力電
流との関係を示すグラフである。 （符号の説明） １……針状対極 ２……中空針の半截物 ４……絶縁層 ５……GOD固定化平板状作用極 ６……平板 ７……固定化GODBRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view showing one embodiment of the glucose sensor of the present invention before being bonded. Further, FIG. 2 is a graph showing the relationship between glucose concentration and output current when this glucose sensor is used. (Explanation of symbols) 1 ... Needle counter electrode 2 ... Hollow needle semi-assembly 4 ... Insulating layer 5 ... GOD-fixed flat plate working electrode 6 ... Plate 7 ... Fixed GOD
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63273130AJP2689531B2 (en) | 1988-10-31 | 1988-10-31 | Glucose sensor |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63273130AJP2689531B2 (en) | 1988-10-31 | 1988-10-31 | Glucose sensor |
| Publication Number | Publication Date |
|---|---|
| JPH02120655A JPH02120655A (en) | 1990-05-08 |
| JP2689531B2true JP2689531B2 (en) | 1997-12-10 |
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63273130AExpired - Fee RelatedJP2689531B2 (en) | 1988-10-31 | 1988-10-31 | Glucose sensor |
| Country | Link |
|---|---|
| JP (1) | JP2689531B2 (en) |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5593852A (en) | 1993-12-02 | 1997-01-14 | Heller; Adam | Subcutaneous glucose electrode |
| US5437999A (en)* | 1994-02-22 | 1995-08-01 | Boehringer Mannheim Corporation | Electrochemical sensor |
| EP1579814A3 (en) | 1996-05-17 | 2006-06-14 | Roche Diagnostics Operations, Inc. | Methods and apparatus for sampling and analyzing body fluid |
| US7828749B2 (en) | 1996-05-17 | 2010-11-09 | Roche Diagnostics Operations, Inc. | Blood and interstitial fluid sampling device |
| US7235056B2 (en) | 1996-05-17 | 2007-06-26 | Amira Medical | Body fluid sampling device and methods of use |
| US20020010406A1 (en) | 1996-05-17 | 2002-01-24 | Douglas Joel S. | Methods and apparatus for expressing body fluid from an incision |
| AU6157898A (en) | 1997-02-06 | 1998-08-26 | E. Heller & Company | Small volume (in vitro) analyte sensor |
| US6071391A (en) | 1997-09-12 | 2000-06-06 | Nok Corporation | Enzyme electrode structure |
| US6103033A (en) | 1998-03-04 | 2000-08-15 | Therasense, Inc. | Process for producing an electrochemical biosensor |
| US6134461A (en) | 1998-03-04 | 2000-10-17 | E. Heller & Company | Electrochemical analyte |
| US9066695B2 (en) | 1998-04-30 | 2015-06-30 | Abbott Diabetes Care 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 |
| US8974386B2 (en) | 1998-04-30 | 2015-03-10 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US8480580B2 (en) | 1998-04-30 | 2013-07-09 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| 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 |
| US8465425B2 (en) | 1998-04-30 | 2013-06-18 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| 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 |
| US6560471B1 (en) | 2001-01-02 | 2003-05-06 | Therasense, Inc. | Analyte monitoring device and methods of use |
| US7310543B2 (en) | 2001-03-26 | 2007-12-18 | Kumetrix, Inc. | Silicon microprobe with integrated biosensor |
| DE10134650B4 (en) | 2001-07-20 | 2009-12-03 | Roche Diagnostics Gmbh | System for taking small amounts of body fluid |
| US20030153026A1 (en) | 2002-01-04 | 2003-08-14 | Javier Alarcon | Entrapped binding protein as biosensors |
| US7381184B2 (en) | 2002-11-05 | 2008-06-03 | Abbott Diabetes Care Inc. | Sensor inserter assembly |
| US7815579B2 (en) | 2005-03-02 | 2010-10-19 | Roche Diagnostics Operations, Inc. | Dynamic integrated lancing test strip with sterility cover |
| EP1578262A4 (en) | 2002-12-31 | 2007-12-05 | Therasense Inc | Continuous glucose monitoring system and methods of use |
| USD914881S1 (en) | 2003-11-05 | 2021-03-30 | Abbott Diabetes Care Inc. | Analyte sensor electronic mount |
| EP1718198A4 (en) | 2004-02-17 | 2008-06-04 | Therasense Inc | Method and system for providing data communication in continuous glucose monitoring and management system |
| 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 |
| US8226891B2 (en) | 2006-03-31 | 2012-07-24 | Abbott Diabetes Care Inc. | Analyte monitoring devices and methods therefor |
| US7620438B2 (en) | 2006-03-31 | 2009-11-17 | Abbott Diabetes Care Inc. | Method and system for powering an electronic device |
| JP4953139B2 (en) | 2006-04-17 | 2012-06-13 | 住友電気工業株式会社 | Biosensor chip |
| JP4924923B2 (en)* | 2006-11-01 | 2012-04-25 | 住友電気工業株式会社 | Sensor cartridge |
| EP2090227A4 (en) | 2006-11-10 | 2010-01-27 | Nat Inst Of Advanced Ind Scien | BIODELECTOR CARTRIDGE, BIOSENSOR DEVICE, SPECIMEN SAMPLING METHOD, METHOD FOR MANUFACTURING BIOSENSOR CARTRIDGE, AND INTEGRATED NEEDLE SENSOR |
| WO2008075651A1 (en) | 2006-12-19 | 2008-06-26 | National Institute Of Advanced Industrial Science And Technology | Biosensor cartridge, method of using biosensor cartridge, biosensor device and needle-integrated sensor |
| US8123686B2 (en) | 2007-03-01 | 2012-02-28 | Abbott Diabetes Care Inc. | Method and apparatus for providing rolling data in communication systems |
| US20100213057A1 (en) | 2009-02-26 | 2010-08-26 | Benjamin Feldman | Self-Powered Analyte Sensor |
| JP6701748B2 (en)* | 2016-01-19 | 2020-05-27 | ヤマハ株式会社 | Strain sensor element |
| US12239463B2 (en) | 2020-08-31 | 2025-03-04 | Abbott Diabetes Care Inc. | Systems, devices, and methods for analyte sensor insertion |
| Publication number | Publication date |
|---|---|
| JPH02120655A (en) | 1990-05-08 |
| Publication | Publication Date | Title |
|---|---|---|
| JP2689531B2 (en) | Glucose sensor | |
| Sternberg et al. | Study and development of multilayer needle-type enzyme-based glucose microsensors | |
| Chen et al. | A biocompatible needle-type glucose sensor based on platinum-electroplated carbon electrode | |
| US4545382A (en) | Sensor for components of a liquid mixture | |
| US20190094169A1 (en) | Enzyme matrices for use with ethylene oxide sterilization | |
| JPS6212847A (en) | Membrane through which liquid and solute can be permeated, manufacture thereof and utilization thereof | |
| JPH05506172A (en) | implantable glucose sensor | |
| US20150122645A1 (en) | Enzyme matrices for biosensors | |
| WO1997015827A9 (en) | Coated wire sensor | |
| WO1997015827A1 (en) | Coated wire sensor | |
| JPH0750058B2 (en) | Enzyme-immobilized electrode and method for producing the same | |
| CN107064261A (en) | A kind of biology sensor and detection method based on glucose dehydrogenase | |
| JP3393361B2 (en) | Biosensor | |
| JPH0231741A (en) | Electrode for immobilizing physiologically active substance | |
| Vadgama | Biosensors: adaptation for practical use | |
| Atta et al. | Nanobiosensor for health care | |
| JPS59164953A (en) | Immobilized enzyme film and manufacture thereof | |
| JP2748516B2 (en) | Glucose sensor | |
| JP2705199B2 (en) | Glucose sensor | |
| Budnikov | Biomedical aspects of electrochemical methods of analysis | |
| JPH0222552A (en) | Stylus electrode | |
| JP2730085B2 (en) | Glucose sensor | |
| JPS59210356A (en) | Triglyceride sensor | |
| JPH11271258A (en) | Measuring system of physiological phenomenon by sensor fusion | |
| JPH0541685Y2 (en) |
| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |