JPH01112148A - Enzyme sensor - Google Patents
Enzyme sensorInfo
- Publication number
- JPH01112148A JPH01112148A JP62269188A JP26918887A JPH01112148A JP H01112148 A JPH01112148 A JP H01112148A JP 62269188 A JP62269188 A JP 62269188A JP 26918887 A JP26918887 A JP 26918887A JP H01112148 A JPH01112148 A JP H01112148A
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- type fet
- film
- sensor
- ion
- 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.)
- Pending
Links
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 38
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 38
- HTXDPTMKBJXEOW-UHFFFAOYSA-N dioxoiridium Chemical compound O=[Ir]=O HTXDPTMKBJXEOW-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910000457 iridium oxide Inorganic materials 0.000 claims abstract description 8
- 230000006698 induction Effects 0.000 claims description 2
- 229910003445 palladium oxide Inorganic materials 0.000 claims description 2
- JQPTYAILLJKUCY-UHFFFAOYSA-N palladium(ii) oxide Chemical group [O-2].[Pd+2] JQPTYAILLJKUCY-UHFFFAOYSA-N 0.000 claims description 2
- 229910001925 ruthenium oxide Inorganic materials 0.000 claims 1
- WOCIAKWEIIZHES-UHFFFAOYSA-N ruthenium(iv) oxide Chemical compound O=[Ru]=O WOCIAKWEIIZHES-UHFFFAOYSA-N 0.000 claims 1
- 239000000758 substrate Substances 0.000 abstract description 5
- 239000007788 liquid Substances 0.000 abstract description 4
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 abstract description 3
- 229910052710 silicon Inorganic materials 0.000 abstract description 3
- 239000010703 silicon Substances 0.000 abstract description 3
- 239000012212 insulator Substances 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 24
- 239000012528 membrane Substances 0.000 description 19
- 150000002500 ions Chemical class 0.000 description 17
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 8
- 239000004202 carbamide Substances 0.000 description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 7
- 239000008103 glucose Substances 0.000 description 7
- 108010046334 Urease Proteins 0.000 description 6
- 238000010586 diagram Methods 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000011810 insulating material Substances 0.000 description 4
- 108010015776 Glucose oxidase Proteins 0.000 description 3
- 239000004366 Glucose oxidase Substances 0.000 description 3
- 229940116332 glucose oxidase Drugs 0.000 description 3
- 235000019420 glucose oxidase Nutrition 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000003100 immobilizing effect Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000015439 Phospholipases Human genes 0.000 description 1
- 108010064785 Phospholipases Proteins 0.000 description 1
- 229910052581 Si3N4 Inorganic materials 0.000 description 1
- 229910021607 Silver chloride Inorganic materials 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- -1 hydrogen ions Chemical class 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 description 1
- BPUBBGLMJRNUCC-UHFFFAOYSA-N oxygen(2-);tantalum(5+) Chemical compound [O-2].[O-2].[O-2].[O-2].[O-2].[Ta+5].[Ta+5] BPUBBGLMJRNUCC-UHFFFAOYSA-N 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- HQVNEWCFYHHQES-UHFFFAOYSA-N silicon nitride Chemical compound N12[Si]34N5[Si]62N3[Si]51N64 HQVNEWCFYHHQES-UHFFFAOYSA-N 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 1
- 229910001415 sodium ion Inorganic materials 0.000 description 1
- 238000004544 sputter deposition Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229910001936 tantalum oxide Inorganic materials 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は酵素の働きにより生体基質の濃度を測定する酵
素サンサに関するものである。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to an enzyme sensor that measures the concentration of a biological substrate through the action of an enzyme.
［従来の技術］
近年、半導体技術及びこの技術を利用したＩＣ技術の進
歩と相まって、ＭＯＳＦＥＴが水素イオン、ナトリウム
イオン等のイオン濃度測定用センサ、あるいは酵素を固
定化し酵素固定化膜を用いた酵素センサ等に広く用いら
れてきている。[Conventional technology] In recent years, coupled with advances in semiconductor technology and IC technology that utilizes this technology, MOSFETs have become popular as sensors for measuring ion concentrations such as hydrogen ions and sodium ions, or as sensors for immobilizing enzymes and using enzyme-immobilized membranes. It has been widely used in sensors, etc.
ＭＯＳＦＥＴは、一般に基板、バリヤー膜及びイオン感
応膜から構成され、イオン感応膜はゲート部に接続され
ている。そして、ＭＯＳＦＥＴを被検液に浸漬すると、
イオン濃度に応じてイオン感応膜の表面電位が変化し、
この電位変化を例えばソースとドレイン間の電流変化と
して測定し、標準溶液での結果を参照することによって
イオン濃度を知ることがで籾る。A MOSFET is generally composed of a substrate, a barrier film, and an ion-sensitive film, and the ion-sensitive film is connected to a gate portion. Then, when the MOSFET is immersed in the test liquid,
The surface potential of the ion-sensitive membrane changes depending on the ion concentration,
The ion concentration can be determined by measuring this potential change, for example, as a current change between the source and drain, and by referring to the results with a standard solution.
また、イオン感応膜上に測定対象物を分解してプロトン
を発生させる酵素を固定化することにより酵素センサと
なる。しかし、ＭＯＳＦＥＴの高いノイズレベルのため
、低周波領域で雑音が高くなる欠点があった。Furthermore, an enzyme sensor can be obtained by immobilizing an enzyme that decomposes an object to be measured and generates protons on an ion-sensitive membrane. However, due to the high noise level of the MOSFET, there is a drawback that the noise becomes high in the low frequency region.
［発明が解決しようとする問題点コ
本発明の目的は、低雑音性を有し、しかも低濃度の被検
液に対して測定可能た酵素センサのを提供することにあ
る。[Problems to be Solved by the Invention] An object of the present invention is to provide an enzyme sensor that has low noise and is capable of measuring a low concentration test liquid.
［問題点を解決するための手段及び作用］上記目的は、
接合型ＦＥＴと、該接合型ＦＥＴのイオン感応部に生成
された導電性層と、該導電性層を被覆する酵素固定層か
らなる酵素センサにより達成される。[Means and actions for solving the problem] The above purpose is to
This is achieved by an enzyme sensor consisting of a junction FET, a conductive layer formed on the ion-sensitive part of the junction FET, and an enzyme immobilization layer covering the conductive layer.
［実施例］
本発明の一実施例の酵素サンサな、第１図（ａ）とＡＡ
’断面図の第１図（ｂ）とＢＢ′断面図の第１図（ｃ）
に示す例を用いて説明する。[Example] Figure 1 (a) and AA of the enzyme sample of one example of the present invention
Figure 1 (b) of 'sectional view' and Figure 1 (c) of BB' cross-sectional view
This will be explained using the example shown in .
本実施例の酵素センサは、ｐ型シリコン基板１上にｐｎ
接合型ＦＥＴが形成され、このｐｎ接合型ＦＥＴより少
し離れた位置に酵素固定化膜３を有するイオン感応膜２
であるイリジウムオキサイドが絶縁材４の上に設けられ
ている。The enzyme sensor of this embodiment has pn on a p-type silicon substrate 1.
An ion-sensitive membrane 2 in which a junction type FET is formed and has an enzyme-immobilized membrane 3 at a position slightly apart from the pn junction type FET.
Iridium oxide is provided on the insulating material 4.
ｐｎ接合型ＦＥＴは、ドレイン部６とソース部７とゲー
ト部５とを有していて、ゲート部５はイオン感応膜２と
接続されている。The pn junction FET has a drain section 6, a source section 7, and a gate section 5, and the gate section 5 is connected to the ion sensitive film 2.
イオン感応膜２としては、イリジウムオキサイド、パラ
ジウムオキサイド、窒化ケイ素、酸化アルミニウム、酸
化タンタルなどが使用できる。Iridium oxide, palladium oxide, silicon nitride, aluminum oxide, tantalum oxide, etc. can be used as the ion-sensitive membrane 2.
イオン感応膜２の上でゲート部５より離間した位置に、
厚さ３００〜１０００人の酵素固定化膜３が設けられて
いる。この酵素固定化膜３は、酵素を含有するとともに
、その酵素をイオン感応膜２上に支持する働きをする。On the ion-sensitive membrane 2, at a position spaced apart from the gate part 5,
An enzyme-immobilized membrane 3 having a thickness of 300 to 1000 membranes is provided. This enzyme-immobilized membrane 3 contains an enzyme and functions to support the enzyme on the ion-sensitive membrane 2.
本実施例に使用される酵素としては、測定対象物を分解
してプロトンを発生させるものならば使用でき、例えば
、ウレアーゼ（尿素検出用）。As the enzyme used in this example, any enzyme that decomposes the substance to be measured and generates protons can be used, such as urease (for urea detection).
グルコースオキシダーゼ（グルコース検出用）。Glucose oxidase (for glucose detection).
ベニシリナーゼ、トリプシン、リパーゼ、ホスホリパー
ゼ、ベプチターゼ等があげられる。Examples include benicillinase, trypsin, lipase, phospholipase, and veptidase.
第１図（ａ）〜（ｃ）の実施例では、絶縁材４の上に酵
素固定化膜３を被覆しないイリジウムオキサイドによる
イオン感応膜を作製し、これを基準電極部８として使用
し、このイオン感応膜とｐｎ接合型ＦＥＴのゲート部５
とを、抵抗１０を介して接続している。このように作製
された酵素センサは、単一で第２図に示す回路を形成す
ることが出来る。In the embodiment shown in FIGS. 1(a) to (c), an ion-sensitive membrane made of iridium oxide without covering the enzyme-immobilized membrane 3 was prepared on the insulating material 4, and this was used as the reference electrode part 8. Ion sensitive membrane and gate part 5 of pn junction type FET
are connected via a resistor 10. A single enzyme sensor produced in this manner can form the circuit shown in FIG. 2.
以下の実施例ではウレアーゼ、グルコースオキシターゼ
について説明する。尚、ここで説明する酵素センサは第
１図（ａ）〜（Ｃ）に示した単一のセンサではなく、接
合型ＦＥＴセンサと基準電極を分離したものである。し
かし、その特性は第１図（ａ）〜（ｃ）に示した最良の
形態と何ら相違しないことは明らかである。In the following examples, urease and glucose oxidase will be explained. Note that the enzyme sensor described here is not the single sensor shown in FIGS. 1(a) to (C), but is one in which a junction type FET sensor and a reference electrode are separated. However, it is clear that its characteristics are no different from the best mode shown in FIGS. 1(a) to (c).
〈実施例１〉
（１）イオン感応膜の作成
絶縁材４上に、スパッタリング法により、イリジウムオ
キサイド膜（厚さ８００人）を作成した。この膜厚は、
２００〜５０００人が好く、好ましくは５００〜１５０
０人であり、特に好ましくは６００〜１０００人である
。これは、薄いと膜抵抗が高いなりすぎ、厚すぎると膜
の剥離等は生じるからである。<Example 1> (1) Creation of ion-sensitive film An iridium oxide film (800 mm thick) was created on the insulating material 4 by sputtering. This film thickness is
200-5000 people, preferably 500-150 people
0 person, particularly preferably 600 to 1000 people. This is because if the film is too thin, the film resistance will be too high, and if it is too thick, the film will peel off.
（２）酵素固定化膜の作成
（１）で作成したイリジウムオキサイド膜上をウレアー
ゼを担持する酵素固定化膜３で被覆した。(2) Creation of enzyme-immobilized membrane The iridium oxide membrane prepared in (1) was coated with enzyme-immobilized membrane 3 that supports urease.
酵素固定化膜３は、
０．２Ｍ（７）ｌ−リス−塩酸緩衝液（ｐＨ８，５）−
１５％牛血清アルブミン溶液
（半井化学薬品株式会社製）に、
ウレアーゼ（東洋紡績株式会社製）
を溶解させた液５μｋを、マイクロシリンジで採取して
酸化イリジウム層の露出面の上に塗布・風乾させた後、
２５容積％のグルタルアルデヒド溶液を０．５μ℃滴下
し、架橋反応によって８００人のウレアーゼ相持の酵素
固定化膜３を成層した。又比較例１．２として、ＭＯＳ
ＦＥＴを用いて同様の尿素センサを作成した。The enzyme-immobilized membrane 3 is made of 0.2M (7) l-Lis-HCl buffer (pH 8.5).
Collect 5 μk of urease (manufactured by Toyobo Co., Ltd.) in a 15% bovine serum albumin solution (manufactured by Hanui Chemical Co., Ltd.) using a microsyringe, apply it on the exposed surface of the iridium oxide layer, and air dry. After letting
A 25 volume % glutaraldehyde solution was added dropwise at 0.5 .mu.C, and an enzyme-immobilized membrane 3 containing 800 ureases was formed by a crosslinking reaction. Also, as Comparative Example 1.2, MOS
A similar urea sensor was created using an FET.
このようにして得られた、尿素センサ２０を用いて、基
準電極２１として銀／塩化銀電極を用いて測定を行なっ
た。測定回路は第２図のように示される。結果を表−１
及び第３図に示す。Using the urea sensor 20 thus obtained, measurements were performed using a silver/silver chloride electrode as the reference electrode 21. The measurement circuit is shown as in FIG. Table 1 shows the results.
and shown in FIG.
表−１
〈実施例２〉
実施例１で示したウレアーゼのかわりにグルコースオキ
シターゼを用いた以外は、実施例１と同様にしてグルコ
ースセンサを作成した。又比較例３．４としてＭＯＳＦ
ＥＴを用いて同様のグルコースセンサを作成した。この
ようにして得られたグルコースセンサを用いて、測定を
行なった。測定回路は第２図に示す回路である。結果を
表−２及び第４図に示す。 表−２
実施例１で用いた尿素センサと比較例１の尿素センサの
雑音特性を測定した結果を第５図に示す。尚、この測定
は、リン酸緩衝液中でスペクトロアナライザーで測定し
た。Table 1 <Example 2> A glucose sensor was produced in the same manner as in Example 1 except that glucose oxidase was used instead of urease shown in Example 1. Also, as comparative example 3.4, MOSF
A similar glucose sensor was created using ET. Measurements were performed using the glucose sensor obtained in this way. The measuring circuit is the circuit shown in FIG. The results are shown in Table 2 and Figure 4. Table 2 The results of measuring the noise characteristics of the urea sensor used in Example 1 and the urea sensor of Comparative Example 1 are shown in FIG. Note that this measurement was performed using a spectroanalyzer in a phosphate buffer.
以上の結果から応答速度は従来のＭＯＳＦＥＴを使用し
たものと同様であるものの、
１）１〜１０１００（／ｄ１）という、低濃度の被検液
に対して測定が可能である。From the above results, although the response speed is the same as that using a conventional MOSFET, 1) It is possible to measure a test liquid with a low concentration of 1 to 10100 (/d1).
２）低周波数領域での雑音特性が優れていることが判明
した。2) It was found that the noise characteristics in the low frequency region are excellent.
更に、ショットキー型ＦＥＴあるいは静電誘導型ＦＥＴ
でも同様の結果が得られ、酵素センサに適用できること
が判明した。尚、本実施例では、尿素センサとグルコー
スセンサを代表して説明したが、他の酵素センサでも同
様の効果があることは明らかである。Furthermore, Schottky type FET or static induction type FET
However, similar results were obtained, indicating that the method can be applied to enzyme sensors. In this example, a urea sensor and a glucose sensor have been described as representatives, but it is clear that other enzyme sensors can have similar effects.
［発明の効果］
本発明により、低周波領域での雑音特性に優れ、低濃度
でも測定可能な酵素センサが得られる。[Effects of the Invention] According to the present invention, an enzyme sensor having excellent noise characteristics in a low frequency region and capable of measuring even low concentrations can be obtained.
第１図（ａ）は本実施例の酵素センサの構成を示す模式
図、
第１図（ｂ）は第１図（ａ）の酵素センサのＡＡ′断面
図、
第１図（ｅ）は第１図（ａ）の酵素センサのＢＢ′断面
図、
第２図は本実施例の酵素センサを試験した測定回路を示
す図、
第３図は本実施例の尿素センサの尿素濃度に対する超電
力を示す図、
第４図は本実施例のグルコースセンサのグルコース濃度
に対する超電力を示す図、
第５図は本実施例及び比較例の雑音特性を示す図である
。
図中、１・・・シリコン基板、２・・・イオン感応膜、
３・・・酵素固定化膜、４・・・絶縁材、５・・・ゲー
ト部、６・・・ドレイン部、７・・・ソース部、８・・
・基準電極部、１０・・・抵抗である。
第２図
第３図FIG. 1(a) is a schematic diagram showing the configuration of the enzyme sensor of this example, FIG. 1(b) is a sectional view taken along line AA′ of the enzyme sensor of FIG. 1(a), and FIG. Figure 1(a) is a BB' cross-sectional view of the enzyme sensor, Figure 2 is a diagram showing the measurement circuit used to test the enzyme sensor of this example, and Figure 3 is a diagram showing the superpower for the urea concentration of the urea sensor of this example. FIG. 4 is a diagram showing superpower versus glucose concentration of the glucose sensor of this example, and FIG. 5 is a diagram showing noise characteristics of this example and a comparative example. In the figure, 1... silicon substrate, 2... ion sensitive film,
3... Enzyme immobilization film, 4... Insulating material, 5... Gate part, 6... Drain part, 7... Source part, 8...
-Reference electrode section, 10...resistance. Figure 2 Figure 3
Claims (3)
に生成された導電性層と、該導電性層を被覆する酵素固
定化層からなることを特徴とする酵素センサ。(1) An enzyme sensor comprising a junction FET, a conductive layer formed on an ion-sensitive portion of the junction FET, and an enzyme immobilization layer covering the conductive layer.
ー型ＦＥＴ、静電誘導型ＦＥＴから選ばれることを特徴
とする特許請求の範囲第１項記載の酵素センサ。(2) The enzyme sensor according to claim 1, wherein the junction type FET is selected from a pn junction type FET, a Schottky type FET, and an electrostatic induction type FET.
れることを特徴とする特許請求の範囲第１項記載の酵素
センサ。(3) The conductive layer is iridium oxide. The enzyme sensor according to claim 1, wherein the enzyme sensor is selected from palladium oxide and ruthenium oxide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62269188A JPH01112148A (en) | 1987-10-27 | 1987-10-27 | Enzyme sensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62269188A JPH01112148A (en) | 1987-10-27 | 1987-10-27 | Enzyme sensor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01112148A true JPH01112148A (en) | 1989-04-28 |
Family
ID=17468905
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62269188A Pending JPH01112148A (en) | 1987-10-27 | 1987-10-27 | Enzyme sensor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01112148A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1035411A1 (en) * | 1999-03-05 | 2000-09-13 | AVL Medical Instruments AG | Electrochemical sensor |
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|---|---|---|---|---|
| JPS61254845A (en) * | 1985-05-08 | 1986-11-12 | Nec Corp | Biosensor and its production |
| JPS62185160A (en) * | 1986-02-10 | 1987-08-13 | Terumo Corp | Biosensor |
-
1987
- 1987-10-27 JP JP62269188A patent/JPH01112148A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61254845A (en) * | 1985-05-08 | 1986-11-12 | Nec Corp | Biosensor and its production |
| JPS62185160A (en) * | 1986-02-10 | 1987-08-13 | Terumo Corp | Biosensor |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1035411A1 (en) * | 1999-03-05 | 2000-09-13 | AVL Medical Instruments AG | Electrochemical sensor |
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