JP2689531B2 - Glucose sensor - Google Patents

Glucose sensor

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Publication number
JP2689531B2
JP2689531B2 JP27313088A JP27313088A JP2689531B2 JP 2689531 B2 JP2689531 B2 JP 2689531B2 JP 27313088 A JP27313088 A JP 27313088A JP 27313088 A JP27313088 A JP 27313088A JP 2689531 B2 JP2689531 B2 JP 2689531B2
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Expired - Fee Related
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JP27313088A
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Japanese (ja)
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JPH02120655A (en )
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正男 後藤
一郎 高津
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エヌオーケー株式会社
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【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。 BACKGROUND OF THE INVENTION [FIELD OF THE INVENTION The present invention relates to a glucose sensor. 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。 More particularly, to glucose sensors amperometric type.

〔従来の技術〕 [Prior art]

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。 In conventional amperometric glucose sensor, it has required two electrodes, one of which is the counter electrode, the one other a working electrode, the working electrode glucose oxidase enzyme (GOD) is It is immobilized. 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。 The principle of this sensor, by quantifying the electrode detection substance occurring in companion no working electrode for reaction with glucose and GOD, in that it can determine the glucose concentration.

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。 Incidentally, as one form of the glucose sensor include but are needle-like structure, in the conventional amperometric glucose sensor was constructed by arranging in parallel the ones flat, such as these two electrodes, for example, a glass plate what is typical, has not yet been put to practical use that the needle electrode which can be used suitably for the measurement, such as insertion to facilitate blood glucose in vivo. 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。 This is believed to have been derived from the difficulty of difficulty and enzyme immobilization of fabrication attributed to the shape itself.

〔発明が解決しようとする課題〕 [Problems that the Invention is to Solve]

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 that such overcome the problems, to provide amperometric type glycose sensor ease of enzyme immobilization and production can be obtained.

〔課題を解決するための手段〕 [Means for Solving the Problems]

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 allowed to achieve the above object, a glucose oxidase enzyme immobilization flat working electrode via an insulating layer on the cut surface of the hollow needle-like counter electrode which is half-cut in the longitudinal direction of the enzyme immobilization face inwardly formed by bonding toward.

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。 The first figures of the drawing, one embodiment of the glucose sensor according to the present invention is shown in perspective view in a state before bonding. 即ち、針状対極1は、一般に外径が約1mm以下の白金、 Namely, needle-like counter electrode 1 is generally an outer diameter of about 1mm or less platinum,
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。 Gold, a half-cut was 2 in the length direction of the hollow needle, such as titanium, the tip portion 3 is beveled to an injection needle so as to be easily inserted. 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。 Then, the cut surface which is half-cut, generally also serves as an adhesive layer insulating layers 4 and 4 ', for example, an epoxy resin adhesive, such as silicone adhesive or glass is applied.

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、 On the other hand, the glucose oxidase enzyme immobilization flat working electrode 5 which is adhered to the hollow needle counter electrode through the insulating layer,
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。 Thickness of about 0.05~1mm about platinum, a flat plate 6, such as titanium, typically that is equal to or less than the length of the counter electrode cutting surface length, the GOD allowed immobilized 7 on one side of a width not equal to the cut surface interval ing.

GODの作用極上への固定化に際しては、膜形成材料、 In the immobilization of GOD to the working electrode, the film-forming material,
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。 For example, cellulose such as cellulose acetate, polyvinyl butyral, synthetic polymeric substances such as polysulfone, or cellulose, chitin, albumin, sodium alginate, agarose, natural polymeric substance such as carrageenan are used, and let them dissolve soluble solvent after preparing a dope solution, a dipping method, dropping method, it is applied onto the working electrode surface by the like spinner method. 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。 Furthermore, can be used as the film forming material, stilbazolium groups as photocrosslinkable groups in the molecule, as aqueous solutions such as water-soluble photo-crosslinkable polymer such as polyvinyl alcohol having a photosensitive group such as Jiaso group.

これらの膜形成材料によって形成される膜上へのGOD GOD onto films formed by these film-forming material
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。 Immobilization, covalent bonding method using glutaraldehyde, carbodiimide and the like, ionic bonding method, performed by the method used adsorption method, etc. In general crosslinking method. あるいは、膜形成材料の溶液中などにGODを混合しておき、 Alternatively, in advance by mixing GOD etc. solution of film-forming material,
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。 It can also be immobilized by these fixing methods during film formation. 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 In this case, when the photo-crosslinkable polymer is used, approximately that of photocrosslinking a wavelength which does not deactivate the GOD 3
50〜450nmの近紫外線による光照射によって行われる。 Performed by the light irradiation by near UV 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、 Determination of glucose by such immobilized GOD is なる反応により、生成したH 2 O 2を作用極での酸化電流値変化として検出する。 By comprising the reaction, detecting the generated H 2 O 2 as an oxidation current value change 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, Further, ferrocene compound is an electron mediator (electron mediator) include ferrocene [bis (cyclopentadienyl) iron (II)] or a derivative thereof, for example 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 1'like with dimethyl ferrocene, after it is vacuum deposited on the working electrode surface, allowed to immobilized by conventional immobilization method used the GOD on the deposition surface, or 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、 Overlaid with like therein dropping method ferrocene compound after immobilizing, この場合には、グルコース1分子が酸化されるのに、2 In this case, although one molecule of glucose is oxidized, 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。 Since the movement of electrons to the working electrode takes place, which can be detected as a current value change.

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。 Such adhesion of the hollow needle-like counter electrode GOD immobilized tabular working electrode is carried out as enzyme immobilization surface is directed inward. 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。 Incidentally, from the counter electrode and the working electrode, it is respectively drawn out lead wire 8, 8 ', at the time of measurement is also used apart from the reference electrode and the electrodes.

〔発明の効果〕 〔Effect of the invention〕

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、 Acicular amperometry glucose sensor according to the present invention is not difficulties there because immobilization of glucose oxidase enzyme is performed on the plate-shaped working electrode surface above,
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 difficulties in manufacturing that attributed to the shape of the needle electrode be not be particularly observed, is provided as a glucose sensor which can be used suitably for the measurement, such as easy blood glucose level was inserted into a living body.

〔実施例〕 〔Example〕

次に、実施例について本発明を説明する。 The following describes the present invention for Example.

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。 It was manufactured glucose sensor embodiment shown in Figure 1 embodiment. 中空針状対極は、内径0.8mm、外径1.0mm、長さ3cm Hollow needle-like counter electrode has an inner diameter 0.8 mm, outer diameter 1.0 mm, length 3cm
の白金中空針を半截し、先端部を斜切したものであり、 And half-cut platinum hollow needle, is obtained by bevel the tip,
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。 The flat working electrode, is used as the dimensions of 1.0 × 2.5 × 0.2 mm, it was immobilized GOD on one surface side.

GODの固定化は、GOD30mgおよび牛血清アルブミン30mg GOD of immobilization, GOD30mg and bovine serum albumin 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 The pH7.0 in 5mM Tris - was dissolved in a maleate buffer 0.4 ml, was added dropwise the 4μ on a flat plate-like working electrode surface, 4 4 ° C.
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。 After standing time, dropwise an acetone solution 4μ there to 1,1'-dimethyl-ferrocene, was conducted by allowing to stand for 4 hours again 4 ° C..

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 working electrode, toward the enzyme immobilization face inward, bonded via a hollow needle-like counter electrode in an epoxy resin adhesive layer applied to the cut surface, 48 hours standing at 4 ° C. 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) Connect these respective electrodes to a potentiostat, a counter electrode - working electrode potential between 50 mV, the working electrode - reference electrode (Ag / AgCl electrode)
間電位200mVとして、グルコースに対する応答を測定した。 As between the potential 200 mV, it was measured the response to glucose. 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。 Measurements, 5mM Tris pH7.0 at 37 ° C. - was carried out using a maleate buffer. 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。 As a result, as shown in the graph of FIG. 2, within the range of glucose concentration 50 to 300 mg / dl, it is a linear correlation between the steady-state value of the response current was observed.

【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。 BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view showing an embodiment of the glucose sensor of the present invention in a state before bonding. 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。 Also, FIG. 2 is a graph showing the relationship between the glucose concentration and the output current in the case of using the glucose sensor. (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GOD (Description of symbols) 1 ...... needle counter electrode 2 ...... hollow needle half-cut was 4 ...... insulating layer 5 ...... GOD immobilized tabular working electrode 6 ...... flat 7 ...... immobilized GOD

Claims (1)

    (57)【特許請求の範囲】 (57) [the claims]
  1. 【請求項1】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両側を内側に向けて接着してなるグルコースセンサ。 1. A glucose sensor comprising adhering toward the glucose oxidase enzyme immobilization flat working electrode via an insulating layer on the cut surface of the hollow needle-like counter electrode which is half-cut in the longitudinal direction on both sides enzyme immobilization inside .
JP27313088A 1988-10-31 1988-10-31 Glucose sensor Expired - Fee Related JP2689531B2 (en)

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Families Citing this family (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
JP3394262B2 (en) 1997-02-06 2003-04-07 イー.ヘラー アンド カンパニー Small volume in vitro analyte sensor
US6071391A (en) 1997-09-12 2000-06-06 Nok Corporation Enzyme electrode structure
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
US9066695B2 (en) 1998-04-30 2015-06-30 Abbott Diabetes Care Inc. Analyte monitoring device and methods of use
US6338790B1 (en) 1998-10-08 2002-01-15 Therasense, Inc. Small volume in vitro analyte sensor with diffusible or non-leachable redox mediator
US6591125B1 (en) 2000-06-27 2003-07-08 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
DE10134650B4 (en) 2001-07-20 2009-12-03 Roche Diagnostics Gmbh System for withdrawing small amounts of body fluid
US20030153026A1 (en) 2002-01-04 2003-08-14 Javier Alarcon Entrapped binding protein as biosensors
US7811231B2 (en) 2002-12-31 2010-10-12 Abbott Diabetes Care Inc. Continuous glucose monitoring system and methods of use
US7815579B2 (en) 2005-03-02 2010-10-19 Roche Diagnostics Operations, Inc. Dynamic integrated lancing test strip with sterility cover
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
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
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 Biosensor cartridge, biosensor device, specimen sampling method, manufacturing method for biosensor cartridge, and needle-integrated sensor
EP2098167A1 (en) 2006-12-19 2009-09-09 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

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