JPH0375555A - Manufacture of biosensor - Google Patents
Manufacture of biosensorInfo
- Publication number
- JPH0375555A JPH0375555A JP1212041A JP21204189A JPH0375555A JP H0375555 A JPH0375555 A JP H0375555A JP 1212041 A JP1212041 A JP 1212041A JP 21204189 A JP21204189 A JP 21204189A JP H0375555 A JPH0375555 A JP H0375555A
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- biosensor
- sensor
- electrode
- polymerization
- 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
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 8
- 108090000790 Enzymes Proteins 0.000 claims abstract description 31
- 102000004190 Enzymes Human genes 0.000 claims abstract description 31
- 230000003100 immobilizing effect Effects 0.000 claims abstract description 5
- 239000000126 substance Substances 0.000 claims description 12
- 229920005597 polymer membrane Polymers 0.000 claims 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 12
- 239000001301 oxygen Substances 0.000 abstract description 12
- 229910052760 oxygen Inorganic materials 0.000 abstract description 12
- 238000006116 polymerization reaction Methods 0.000 abstract description 11
- 239000000758 substrate Substances 0.000 abstract description 9
- 238000005259 measurement Methods 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 5
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 229920002521 macromolecule Polymers 0.000 abstract 2
- 239000000463 material Substances 0.000 abstract 2
- 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 21
- 239000008103 glucose Substances 0.000 description 21
- 229940088598 enzyme Drugs 0.000 description 19
- 239000000243 solution Substances 0.000 description 10
- 125000004122 cyclic group Chemical group 0.000 description 7
- 230000027756 respiratory electron transport chain Effects 0.000 description 7
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 6
- 238000010586 diagram Methods 0.000 description 4
- OXHNLMTVIGZXSG-UHFFFAOYSA-N 1-Methylpyrrole Chemical compound CN1C=CC=C1 OXHNLMTVIGZXSG-UHFFFAOYSA-N 0.000 description 3
- 108090000854 Oxidoreductases Proteins 0.000 description 3
- 102000004316 Oxidoreductases Human genes 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 235000011164 potassium chloride Nutrition 0.000 description 3
- 239000001103 potassium chloride Substances 0.000 description 3
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 description 2
- 108010015776 Glucose oxidase Proteins 0.000 description 2
- 239000004366 Glucose oxidase Substances 0.000 description 2
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 2
- 229910021607 Silver chloride Inorganic materials 0.000 description 2
- 229940116332 glucose oxidase Drugs 0.000 description 2
- 235000019420 glucose oxidase Nutrition 0.000 description 2
- 229910052709 silver Inorganic materials 0.000 description 2
- 239000004332 silver Substances 0.000 description 2
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 1
- 102000004674 D-amino-acid oxidase Human genes 0.000 description 1
- 108010003989 D-amino-acid oxidase Proteins 0.000 description 1
- 108010042687 Pyruvate Oxidase Proteins 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 229920001940 conductive polymer Polymers 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 229910001873 dinitrogen Inorganic materials 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920000128 polypyrrole Polymers 0.000 description 1
- 229910052594 sapphire Inorganic materials 0.000 description 1
- 239000010980 sapphire Substances 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 238000010408 sweeping Methods 0.000 description 1
Landscapes
- Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、基質濃度を迅速・簡便に測定できるバイオセ
ンサの製造方法に関する。DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to a method for manufacturing a biosensor that can quickly and easily measure substrate concentration.
（従来の技術）
溶液中の物質の濃度を測定するために、その物質を基質
とする酵素を電極表面に固定化したバイオセンサの作製
が試みられている。グルコースオキシダーゼ（ＣＯＤ）
等の酸化酵素は、酸素を消費して過酸化水素を生成する
。この酵素反応による酸素の減少量や過酸化水素の増加
量から基質濃度を算出する、最も単純な原理に基づくバ
イオセンサは数多く作製されている。近年、酸素の代わ
りに、酵素と電極との間の電子移動を媒介する物質、メ
ゾイエイタ−をバイオセンサに応用した例が報告されて
いる。例えば、ＧＯＤとともにメゾイエイタ−である１
、１′−ジメチルフェロセンを固定化したグルコースセ
ンサが報告されている（アナリテイカル・ケミストリー
５６．６６７−６７１ （１９８４））。(Prior Art) In order to measure the concentration of a substance in a solution, attempts have been made to create a biosensor in which an enzyme using the substance as a substrate is immobilized on the surface of an electrode. Glucose oxidase (COD)
Oxidases such as oxidases consume oxygen and produce hydrogen peroxide. Many biosensors have been created based on the simplest principle, which calculates the substrate concentration from the amount of decrease in oxygen or increase in hydrogen peroxide due to this enzymatic reaction. In recent years, examples have been reported in which mesoieters, which are substances that mediate electron transfer between enzymes and electrodes, are applied to biosensors instead of oxygen. For example, 1 which is a mesoiator along with GOD
, a glucose sensor in which 1'-dimethylferrocene is immobilized has been reported (Analytical Chemistry 56.667-671 (1984)).
一方、電解重合性物質によって酵素を固定化したバイオ
センサについては、ポリピロールを用いてグルコースオ
キシダーゼを固定化したグルコースセンサが作製されて
いる（アナリティカル・ケミストリー５８．２９７９〜
２９８３ （１９８６））。On the other hand, regarding a biosensor in which an enzyme is immobilized using an electrolytically polymerizable substance, a glucose sensor in which glucose oxidase is immobilized using polypyrrole has been produced (Analytical Chemistry 58.2979~
2983 (1986)).
（発明が解決しようとする課題）
酸素を必要とするバイオセンサは、試料中の溶存酸素濃
度が一定である必要があること、酸素不足によって高濃
度の基質の測定ができないこと等の短所がある。メゾイ
エイタ−用いたバイオセンサにほこのような欠点はない
が、メゾイエイタ−の固定化が難しい、メゾイエイタ−
のセンサからの脱離によって寿命が短いという問題が生
じる。(Problems to be solved by the invention) Biosensors that require oxygen have disadvantages such as the need for a constant dissolved oxygen concentration in the sample and the inability to measure highly concentrated substrates due to lack of oxygen. . Although biosensors using mesoiators do not have such drawbacks, it is difficult to immobilize mesoiators, and mesoiators are difficult to immobilize.
Detachment from the sensor causes the problem of short lifespan.
メゾイエイタ−により媒介なしに酵素と電極間の電子移
動が実現できれば、このような問題を解決できるが、従
来の固定化方法では、酵素と電極間に直接電子移動がお
こるようなバイオセンサを作製することはできない。If electron transfer between the enzyme and the electrode could be achieved without an intermediary using a mesoiator, this problem could be solved, but conventional immobilization methods make it difficult to create a biosensor in which electron transfer occurs directly between the enzyme and the electrode. It is not possible.
本発明の目的は、測定に酸素やメゾイエイタ−を必要と
しない、酸素と電極間の直接電子移動がおこるようなバ
イオセンサを提供することにある。An object of the present invention is to provide a biosensor that does not require oxygen or a mesoiator for measurement and allows direct electron transfer between oxygen and electrodes.
（課題を解決するための手段）
本発明は、電解重合性物質と酵素を含有する溶液中で電
解重合をおこなって酵素を固定化し、バイオセンサを作
製する際、電解重合時の溶液温度を４５°Ｃ以上にする
ことを特徴としている。(Means for Solving the Problems) The present invention immobilizes enzymes by performing electropolymerization in a solution containing an electropolymerizable substance and an enzyme, and when producing a biosensor, the solution temperature during electropolymerization is set to 45%. It is characterized by being heated to over °C.
（作用）
このような方法で酵素を固定化したバイオセンサは、酸
素やメゾイエイタ−がなくとも基質濃度に依存する電流
応答を与えた。このことは、電解重合時に酵素の高次構
造が可逆的に変化して、導電性高分子が酵素の活性中心
近傍に形成されて、酵素と電極との直接電子移動が可能
になったためと推測される。このバイオセンサは、酵素
を必要とするバイオセンサに比べて、酸素濃度の制約が
ないので、より高濃度の基質を測定できる。また、測定
のためにセンサに印加する電位が低いので、他物質によ
るセンサ反応への妨害が少ない。(Function) The biosensor in which the enzyme was immobilized using this method gave a current response that depended on the substrate concentration even in the absence of oxygen or mesoiator. We speculate that this is because the higher-order structure of the enzyme changes reversibly during electropolymerization, and a conductive polymer is formed near the active center of the enzyme, allowing direct electron transfer between the enzyme and the electrode. be done. Compared to biosensors that require enzymes, this biosensor is not limited by oxygen concentration, so it can measure substrates at higher concentrations. Furthermore, since the potential applied to the sensor for measurement is low, there is less interference with the sensor reaction by other substances.
さらに、メゾイエイタ−を使用していないので、メゾイ
エイタ−の脱離によるセンサの劣化がない。Furthermore, since no mesoiator is used, there is no deterioration of the sensor due to detachment of the mesoiator.
（比較例）
酵素としてＧＯＤ、電解重合性物質としてＮ−メチルピ
ロールを用いて本発明に基づくグルコースセンサを作製
した。センサのベースには、特願昭６３−２８２７２１
に示した方法で作成した、サファイア基板上に金を蒸着
した電極を用いた。この電極の作用極の面積は０．１８
５ｍｍであった。この電極を０．１Ｍ硫酸ナトリウムに
浸漬、電極電位−１，５〜１．５Ｖ ｖｓ、 Ａｇ／Ａ
ｇＣ１，掃引速度２００ｍＶ／秒で６分間電位掃引をお
こない、電極を清浄にした。１０ｍｇ／ｍｌ ＧＯＤ、
０．１ＭＮ−メチルビロールを含む０．１Ｍ塩化カリウ
ム水溶液中で、電極に０．８Ｖ ｖｓ、 Ａｇ／ＡｇＣ
１の電位を掛けて２．５ｍＣ通電し、定電位電解重合に
よって酵素固定化膜を形成した。電解重合の際、溶液を
２５°Ｃにして作製した。上記のようにして作製したセ
ンサのサイクリックポルタモグラムを測定した。測定装
置の構成を第１図に示す。図中１はグルコースセンサ及
び対極、２は銀・塩化銀参照電極、３はグルコースを含
む０．１Ｍ塩化カリウム溶液、４はポテンシオスタット
及びファンクションジェネレータ、５は記録計である。(Comparative Example) A glucose sensor based on the present invention was produced using GOD as an enzyme and N-methylpyrrole as an electrolytically polymerizable substance. The base of the sensor is based on the patent application No. 63-282721.
An electrode with gold vapor-deposited on a sapphire substrate was used, which was created using the method shown in . The area of the working electrode of this electrode is 0.18
It was 5 mm. This electrode was immersed in 0.1M sodium sulfate, electrode potential -1.5 to 1.5V vs. Ag/A
gC1, a potential sweep was performed for 6 minutes at a sweep rate of 200 mV/sec to clean the electrode. 10mg/ml GOD,
In a 0.1 M potassium chloride aqueous solution containing 0.1 M N-methylpyrrole, 0.8 V vs. Ag/AgC at the electrode.
A potential of 1 was applied and a current of 2.5 mC was applied to form an enzyme-immobilized membrane by potentiostatic electrolytic polymerization. During electrolytic polymerization, the solution was prepared at 25°C. A cyclic portammogram of the sensor produced as described above was measured. The configuration of the measuring device is shown in Figure 1. In the figure, 1 is a glucose sensor and a counter electrode, 2 is a silver/silver chloride reference electrode, 3 is a 0.1M potassium chloride solution containing glucose, 4 is a potentiostat and a function generator, and 5 is a recorder.
測定溶液は測定前に窒素ガスを吹き込んで溶存酸素を除
き、溶液温度は３５°Ｃとした。サイクリックポルタモ
グラムは０〜−ＩＶ ｖｓ、 Ａｇ／ＡｇＣ１の電位を
５０ｍＶ／秒で掃引して測定した。第２図は、測定した
サイクリックポルタモグラムである。図中破線はグルコ
ースがない場合、実線は４０ｍＭグルコースを添加した
場合のポルタモグラムである。Before measurement, nitrogen gas was blown into the measurement solution to remove dissolved oxygen, and the solution temperature was 35°C. The cyclic portamogram was measured by sweeping the potential of 0 to -IV vs. Ag/AgC1 at 50 mV/sec. FIG. 2 shows the measured cyclic portamogram. In the figure, the dashed line is the portamogram in the absence of glucose, and the solid line is the portamogram in the case where 40 mM glucose is added.
−〇、５Ｖ ｖｓ、 Ａｇ／ＡｇＣ１付近にＣＯＤに由
来するピーク電流が観測されるが、グルコースを添加し
ても、ボルタモダラムに変化はなかった。即ち、この条
件で作製したセンサは、活性を有したＧＯＤと電極との
間の電子移動が起こらず、グルコースセンサとして機能
しなかった。-〇, 5V vs. A peak current derived from COD was observed near Ag/AgC1, but there was no change in the voltamodalum even when glucose was added. That is, in the sensor produced under these conditions, electron transfer between the active GOD and the electrode did not occur, and the sensor did not function as a glucose sensor.
（実施例）
前記センサの作製条件において、電解重合の際の溶液を
５０°Ｃにした。第３図はこのセンサのグルコースがな
い場合（破線〉と４０ｍＭグルコースを添加した場合（
実線）のサイクルポルタモグラムである。(Example) Under the manufacturing conditions of the sensor, the temperature of the solution during electrolytic polymerization was 50°C. Figure 3 shows this sensor in the absence of glucose (dashed line) and in the presence of 40mM glucose (dotted line).
This is a cycle portammogram of solid line).
比較例の場合と異なり、４０ｍＭグルコースを加えるこ
とによって一〇、５Ｖ ｖｓ、 Ａｇ／ＡｇＣ１付近の
酸化ピーク電流が増加した。Unlike the case of the comparative example, the oxidation peak current around 10.5 V vs. Ag/AgC1 increased by adding 40 mM glucose.
このセンサのグルコースに対する電流応答を測定した。The current response of this sensor to glucose was measured.
装置はサイクリックポルタモグラム測定の場合と同様で
あり、−〇、３Ｖ ｖｓ、 Ａｇ／ＡｇＣ１の一定電位
をセンサに印加したときの、グルコース添加による電流
の変化を記録した。印加電位−〇、３Ｖ ｖｓ。The apparatus was the same as that for cyclic portammogram measurement, and when a constant potential of -0, 3 V vs. Ag/AgC1 was applied to the sensor, the change in current due to glucose addition was recorded. Applied potential -〇, 3V vs.
Ａｇ／ＡｇＣ１は、従来のグルコースセンサで、過酸化
水素を検出する電位０．５Ｖ ｖｓ、 Ａｇ／ＡｇＣ１
に比べて負電位側にあり、高い電位で酸化される物質に
よる電流応答の妨害が小さい。電流応答のグルコース濃
度依存性を第４図に示した。従来のセンサに比べて高濃
度の１５０ｍＭまでのグルコースが検出できた。Ag/AgC1 is a conventional glucose sensor, and the potential for detecting hydrogen peroxide is 0.5V vs. Ag/AgC1
It is on the negative potential side compared to , and there is less interference with the current response by substances that are oxidized at high potentials. The glucose concentration dependence of the current response is shown in FIG. It was able to detect higher concentrations of glucose up to 150mM compared to conventional sensors.
酵素と電極が直接電子移動をおこなうバイオセンサを作
製するための、電解重合溶液の温度は、用いる酵素や電
解重合性物質の種類や濃度、重合電位によって異なるが
、酵素の高次構造に影響を与える温度である４５°Ｃ以
上が適している。The temperature of the electrolytic polymerization solution used to create a biosensor in which electrons are transferred directly between the enzyme and the electrode varies depending on the type and concentration of the enzyme and electrolytically polymerizable substance used, as well as the polymerization potential, but it does not affect the higher-order structure of the enzyme. A temperature of 45°C or higher is suitable.
なお、上記の二つの実施例では、酵素としてＣＯＤを用
いたが、その他にも、Ｄ−アミノ酸オキシダーゼ、ピル
ビン酸オキシダーゼなど、補酵素としてフラビンアデニ
ジヌクレエチドを有する酸化酵素であればよい。また、
電解重合性物質としては、Ｎ−メチルビロールの他に、
ピロール、アニリンやその誘導体も利用できる。In the above two examples, COD was used as the enzyme, but any other oxidase having flavin adenidine nucleotide as a coenzyme may be used, such as D-amino acid oxidase and pyruvate oxidase. Also,
In addition to N-methylpyrrole, electrolytically polymerizable substances include
Pyrrole, aniline and their derivatives can also be used.
（発明の効果）
本発明は、４５℃以上で酵素を電解重合高分子で固定化
することで、酵素と電極の間の直接電子移動に基づくバ
イオセンサの作製を可能にする。その結果、メゾイエイ
タ−固定化が不要で、他物質の妨害が小さく、高濃度側
まで定量範囲が拡大したバイオセンサが得られた。(Effects of the Invention) The present invention enables the production of a biosensor based on direct electron transfer between an enzyme and an electrode by immobilizing an enzyme with an electrolytically polymerized polymer at 45° C. or higher. As a result, a biosensor was obtained that did not require mesoiator immobilization, had little interference from other substances, and had a quantitative range extended to high concentrations.
第１図は、各実施例におけるセンサのサイクリックポル
タモグラム及びグルコース濃度に対する電流応答の測定
装置を示す図、第２図は比較例におけるセンサのサイク
リックポルタモグラムを表わす図、第３図は実施例にお
けるセンサのサイクリックポルタモグラムを表わす図、
第４図は実施例におけるセンサ電流応答のグルコース濃
度依存性を示す図である。
図中１はグルコースセンサ及び対極、２は銀・塩化銀参
照電極、３はグルコースを含む０．１Ｍ塩化カリウム溶
液、４はポテンシオスタット及びファンクションジェネ
レータ、５は記録計である。FIG. 1 is a diagram showing the cyclic portamogram of the sensor in each example and a device for measuring current response to glucose concentration, FIG. 2 is a diagram showing the cyclic portammogram of the sensor in the comparative example, and FIG. is a diagram showing a cyclic portamogram of the sensor in the example,
FIG. 4 is a diagram showing the glucose concentration dependence of the sensor current response in the example. In the figure, 1 is a glucose sensor and a counter electrode, 2 is a silver/silver chloride reference electrode, 3 is a 0.1M potassium chloride solution containing glucose, 4 is a potentiostat and a function generator, and 5 is a recorder.
Claims (1)
合を行うことによって、酵素を前記重合膜に固定化する
工程を備えたバイオセンサの製造方法において、電解重
合時の溶液温度を４５℃以上にして作製することを特徴
とするバイオセンサの製造方法。(1) In a method for producing a biosensor comprising a step of immobilizing an enzyme on the polymer membrane by performing electropolymerization in a solution containing an electropolymerizable substance and an enzyme, the solution temperature during electropolymerization is set to 45%. A method for producing a biosensor, characterized in that the biosensor is produced at a temperature of at least ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1212041A JPH0375555A (en) | 1989-08-16 | 1989-08-16 | Manufacture of biosensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1212041A JPH0375555A (en) | 1989-08-16 | 1989-08-16 | Manufacture of biosensor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0375555A true JPH0375555A (en) | 1991-03-29 |
Family
ID=16615891
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1212041A Pending JPH0375555A (en) | 1989-08-16 | 1989-08-16 | Manufacture of biosensor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0375555A (en) |
-
1989
- 1989-08-16 JP JP1212041A patent/JPH0375555A/en active Pending
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| Publication | Publication Date | Title |
|---|---|---|
| Geise et al. | Electropolymerized 1, 3-diaminobenzene for the construction of a 1, 1′-dimethylferrocene mediated glucose biosensor | |
| Li et al. | A cholesterol biosensor based on entrapment of cholesterol oxidase in a silicic sol‐gel matrix at a prussian blue modified electrode | |
| Rahman et al. | The biosensor based on the pyruvate oxidase modified conducting polymer for phosphate ions determinations | |
| Foulds et al. | Enzyme entrapment in electrically conducting polymers. Immobilisation of glucose oxidase in polypyrrole and its application in amperometric glucose sensors | |
| EP1194585B1 (en) | Amperometric sensor | |
| HabermuÈller et al. | An oxygen‐insensitive reagentless glucose biosensor based on osmium‐complex modified polypyrrole | |
| Palmisano et al. | A disposable, reagentless, third-generation glucose biosensor based on overoxidized poly (pyrrole)/tetrathiafulvalene− tetracyanoquinodimethane composite | |
| D O’Neill et al. | Comparisons of platinum, gold, palladium and glassy carbon as electrode materials in the design of biosensors for glutamate | |
| Vostiar et al. | Amperometric urea biosensor based on urease and electropolymerized toluidine blue dye as a pH-sensitive redox probe | |
| Ghica et al. | A glucose biosensor using methyl viologen redox mediator on carbon film electrodes | |
| Koopal et al. | Glucose sensor utilizing polypyrrole incorporated in tract-etch membranes as the mediator | |
| Soylemez et al. | Electrochemical and optical properties of a conducting polymer and its use in a novel biosensor for the detection of cholesterol | |
| Zhou et al. | Development of an amperometric biosensor based on covalent immobilization of tyrosinase on a boron-doped diamond electrode | |
| Gobi et al. | Layer-by-layer construction of an active multilayer enzyme electrode applicable for direct amperometric determination of cholesterol | |
| Roy et al. | Vertically aligned carbon nanotube probes for monitoring blood cholesterol | |
| US20130001102A1 (en) | Process for the preparation of modified electrodes, electrodes prepared with said process, and enzymatic biosensors comprising said electrodes | |
| Barsan et al. | New CNT/poly (brilliant green) and CNT/poly (3, 4-ethylenedioxythiophene) based electrochemical enzyme biosensors | |
| Serafín et al. | Electrochemical biosensor for creatinine based on the immobilization of creatininase, creatinase and sarcosine oxidase onto a ferrocene/horseradish peroxidase/gold nanoparticles/multi-walled carbon nanotubes/Teflon composite electrode | |
| Chen et al. | Glucose biosensor based on three dimensional ordered macroporous self-doped polyaniline/Prussian blue bicomponent film | |
| Bu et al. | NAD (P) H sensors based on enzyme entrapment in ferrocene-containing polyacrylamide-based redox gels | |
| Gros et al. | Use of polypyrrole film containing Fe (CN) 63− as pseudo-reference electrode: application for amperometric biosensors | |
| Huang et al. | A highly-sensitive L-lactate biosensor based on sol-gel film combined with multi-walled carbon nanotubes (MWCNTs) modified electrode | |
| Voitechovič et al. | Development of label-free impedimetric platform based on new conductive polyaniline polymer and three-dimensional interdigitated electrode array for biosensor applications | |
| Xu et al. | Glucose biosensors prepared by electropolymerization of p-chlorophenylamine with and without Nafion | |
| Sanz et al. | Disposable superoxide dismutase biosensors based on gold covered polycaprolactone fibers for the detection of superoxide in cell culture media |