JPH0328752A - Enzyme immobilized electrode and its production - Google Patents
Enzyme immobilized electrode and its productionInfo
- Publication number
- JPH0328752A JPH0328752A JP1164797A JP16479789A JPH0328752A JP H0328752 A JPH0328752 A JP H0328752A JP 1164797 A JP1164797 A JP 1164797A JP 16479789 A JP16479789 A JP 16479789A JP H0328752 A JPH0328752 A JP H0328752A
- Authority
- JP
- Japan
- Prior art keywords
- film
- electrode
- enzyme
- immobilized
- membrane
- 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.)
- Granted
Links
- 238000004519 manufacturing process Methods 0.000 title claims description 12
- 108090000790 Enzymes Proteins 0.000 title abstract description 25
- 102000004190 Enzymes Human genes 0.000 title abstract description 25
- 239000000126 substance Substances 0.000 claims abstract description 20
- 230000002452 interceptive effect Effects 0.000 claims abstract description 17
- 239000004020 conductor Substances 0.000 claims abstract description 7
- 239000003906 humectant Substances 0.000 claims description 15
- 230000035945 sensitivity Effects 0.000 abstract description 15
- 238000005259 measurement Methods 0.000 abstract description 10
- 238000004132 cross linking Methods 0.000 abstract description 9
- 238000006243 chemical reaction Methods 0.000 abstract description 7
- 239000007788 liquid Substances 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 230000008859 change Effects 0.000 abstract description 3
- 238000001514 detection method Methods 0.000 abstract description 3
- 239000000203 mixture Substances 0.000 abstract description 2
- 239000012488 sample solution Substances 0.000 abstract description 2
- 230000007547 defect Effects 0.000 abstract 1
- 239000012528 membrane Substances 0.000 description 58
- 239000000243 solution Substances 0.000 description 22
- 229940088598 enzyme Drugs 0.000 description 19
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 12
- 102000008186 Collagen Human genes 0.000 description 11
- 108010035532 Collagen Proteins 0.000 description 11
- 229920001436 collagen Polymers 0.000 description 11
- 239000012298 atmosphere Substances 0.000 description 10
- 239000003431 cross linking reagent Substances 0.000 description 10
- 230000000052 comparative effect Effects 0.000 description 9
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 8
- 239000002585 base Substances 0.000 description 8
- 108010010803 Gelatin Proteins 0.000 description 7
- 229920000159 gelatin Polymers 0.000 description 7
- 239000008273 gelatin Substances 0.000 description 7
- 235000019322 gelatine Nutrition 0.000 description 7
- 235000011852 gelatine desserts Nutrition 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 239000011159 matrix material Substances 0.000 description 6
- 229910052697 platinum Inorganic materials 0.000 description 6
- 108010015776 Glucose oxidase Proteins 0.000 description 5
- 239000004366 Glucose oxidase Substances 0.000 description 5
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 5
- 229940116332 glucose oxidase Drugs 0.000 description 5
- 235000019420 glucose oxidase Nutrition 0.000 description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 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 4
- 239000008103 glucose Substances 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 102000009027 Albumins Human genes 0.000 description 3
- 108010088751 Albumins Proteins 0.000 description 3
- ZNZYKNKBJPZETN-WELNAUFTSA-N Dialdehyde 11678 Chemical compound N1C2=CC=CC=C2C2=C1[C@H](C[C@H](/C(=C/O)C(=O)OC)[C@@H](C=C)C=O)NCC2 ZNZYKNKBJPZETN-WELNAUFTSA-N 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 229920000083 poly(allylamine) Polymers 0.000 description 3
- 239000004909 Moisturizer Substances 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 229920001222 biopolymer Polymers 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 230000001333 moisturizer Effects 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 229920003169 water-soluble polymer Polymers 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 1
- 108010089254 Cholesterol oxidase Proteins 0.000 description 1
- 102000012422 Collagen Type I Human genes 0.000 description 1
- 108010022452 Collagen Type I Proteins 0.000 description 1
- 206010067482 No adverse event Diseases 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 229910010293 ceramic material Inorganic materials 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 239000007772 electrode material Substances 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 230000003533 narcotic effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 229940116269 uric acid Drugs 0.000 description 1
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
この発明は、グルコース等の基質検出用の電極などとし
て用いられる酵素固定化電極（「酵素電極」とも言う）
およびその製造方法に関する。[Detailed Description of the Invention] [Industrial Application Field] This invention relates to an enzyme-immobilized electrode (also referred to as an "enzyme electrode") used as an electrode for detecting a substrate such as glucose.
and its manufacturing method.
酔素を白金等の電極本体（または電極素材。以下同様）
表面上に固定化した酵素固定化電極として、ゼラチンお
よび架橋剤としてのグルタルアルデヒドを含む希薄な酵
素溶液を白金電極に塗布し、電極本体表面上で製膜を行
うとともに、酵素を共有結合的に固定化して電極本体表
面上に酵素固定化膜を形成したものがある。ここで、ゼ
ラチンは、酵素固定化膜のマトリソクス威分となるもの
であって、希薄な．酵素溶液と架橋剤としてのグルタル
アルデヒドだけでは架橋反応により得られる酵素固定化
膜の強度が弱いので、膜強度を高めるために使用される
。たとえば、グルコースオキシダーゼの固定化膜では、
酵素に対して５〜１０倍程度のゼラチンを加えてグルタ
ルアルデヒドで架橋させて製膜するようにしている。Place the intoxicant on the electrode body (or electrode material; the same applies hereinafter) made of platinum, etc.
As an electrode with an enzyme immobilized on the surface, a dilute enzyme solution containing gelatin and glutaraldehyde as a crosslinking agent is applied to a platinum electrode, a film is formed on the surface of the electrode body, and the enzyme is covalently bonded to the platinum electrode. Some enzymes are immobilized to form an enzyme-immobilized film on the surface of the electrode body. Here, gelatin serves as the matrix for the enzyme-immobilized membrane, and is dilute. Since the strength of the enzyme-immobilized membrane obtained by the crosslinking reaction is weak when only the enzyme solution and glutaraldehyde as a crosslinking agent are used, the enzyme solution is used to increase the strength of the membrane. For example, in a glucose oxidase immobilized membrane,
The membrane is formed by adding gelatin in an amount of 5 to 10 times the amount of enzyme and crosslinking with glutaraldehyde.
このような酵素固定化電極を用いるに際しては、試料溶
液中における被測定物質以外の多種多様の威分の妨害か
ら酵素固定化電極を保護する必要がある。そのため、電
極本体表面と酵素固定化膜の間などに妨害物質を除去す
る妨害物質除去膜（以下「妨害除去膜」と言う）を形威
した妨害不感型酵素固定化電極が提案されている。When using such an enzyme-immobilized electrode, it is necessary to protect the enzyme-immobilized electrode from interference from a wide variety of substances other than the substance to be measured in the sample solution. Therefore, an interference-insensitive enzyme-immobilized electrode has been proposed that has an interference-removing membrane (hereinafter referred to as "interference-removal membrane") for removing interfering substances between the surface of the electrode body and the enzyme-immobilized membrane.
たとえば、特開昭６２−８８９５３号公報では、電極本
体表面にグルコースオキシダーゼの固定化膜を設けたも
のを、ビロールおよびＮａ Ｃｆｆを含む電解重合液中
に浸して電解重合処理を行い、前記固定化膜の上に電解
重合膜を備えた酵素固定化電極が記載されている。同電
解重合膜が妨害除去膜である。また、特開昭６３−３０
４１５０翼一公報では、電極本体表面に内側膜、中間膜
および外側膜をこの順に設けた酵素固定化電極が記載さ
れている。中間膜は、グルコースオキシダーゼの固定膜
である。外側膜は、ポリカーボネー１・フィルムであり
、未希釈の血液または血清中のグルコース分子に比べて
高分子物質の除去を行う。内側膜は、酢酸セルロースか
らなる膜であり、ボリヵーボ不−１−フィルムで除去さ
れなかった妨書物質（たとえば、アスコルビン酸および
尿酸）を除去する。For example, in Japanese Patent Application Laid-Open No. 62-88953, an electrode body with a glucose oxidase immobilized membrane provided on the surface thereof is immersed in an electrolytic polymerization solution containing virol and Na Cff to undergo electrolytic polymerization treatment, and the immobilized membrane is An enzyme-immobilized electrode with an electropolymerized membrane on top of the membrane is described. This electrolytically polymerized membrane is an interference removal membrane. Also, JP-A-63-30
Publication No. 4150 Tsubasa describes an enzyme-immobilized electrode in which an inner membrane, an intermediate membrane, and an outer membrane are provided in this order on the surface of the electrode body. The intermediate membrane is a fixed membrane for glucose oxidase. The outer membrane is a polycarbonate 1 film and provides removal of macromolecular substances relative to glucose molecules in undiluted blood or serum. The inner membrane is a membrane made of cellulose acetate and removes interfering substances (eg, ascorbic acid and uric acid) not removed by the polycarbo-1-film.
電極本体表面上で製膜された妨害除去膜を備えた妨害不
感型酵素固定化電極では、妨害除去膜が乾燥に弱いため
、湿度の変化により膜の収縮が生し、水溶液中での測定
および乾燥を繰り返セば、電極本体表面から妨害除去膜
および酵素固定化膜が剥離し、検出感度が低下するなど
して使用できなくなるという欠点がある。In interference-insensitive enzyme-immobilized electrodes that have an interference removal membrane formed on the surface of the electrode body, the interference removal membrane is susceptible to drying, so the membrane shrinks due to changes in humidity, making measurements in aqueous solutions and If drying is repeated, the interference removal film and the enzyme immobilization film will peel off from the surface of the electrode body, resulting in a decrease in detection sensitivity and the like, making it unusable.
また、第３図（ａ）にみるように、酵素を固定化してい
ない妨害除去膜５は、湿度が低ずぎると、電極本体ｌよ
り剥離するため、酵素の固定化作業をたとえば湿度８０
％以上の調湿器内で行わなければならない。Furthermore, as shown in FIG. 3(a), if the humidity is too low, the interference removal film 5 on which the enzyme is not immobilized will peel off from the electrode body l.
% or more must be carried out in a humidifier.
この発明は、このような事情に鑑みてなされたものであ
って、電極本体からの妨害除去膜の剥離が生しにくく、
出力感度の初期変動の小さい酵素固定化電極を提供する
ことを第１の課題とする。This invention was made in view of the above circumstances, and has a structure that prevents the interference removal film from peeling off from the electrode body.
The first objective is to provide an enzyme-immobilized electrode with small initial fluctuations in output sensitivity.
さらに、この発明は、妨害除去膜の上に酵素固定化膜を
形或する場合に、雰囲気の調湿を行わなくてもよい酵素
固定化電極の製造方法を提供することを第２の課題とす
る。Furthermore, a second object of the present invention is to provide a method for manufacturing an enzyme-immobilized electrode that does not require the humidity of the atmosphere when an enzyme-immobilized membrane is formed on an interference removal membrane. do.
発明者らは、妨害除去膜の電極本体からの剥離を防いだ
り、雰囲気の調湿を行わずに妨害除去膜の上に酵素固定
化膜を形戒したりするための手段を検討した。その結果
、妨害除去膜自体に保湿性を持たせればよいことを見出
し、この発明を完成させた。The inventors investigated means for preventing the interference removal membrane from peeling off from the electrode body and for forming an enzyme-immobilized membrane on the interference removal membrane without controlling the humidity of the atmosphere. As a result, they discovered that the interference removal film itself should have moisture retention properties, and completed this invention.
したがって、上記第１の課題を解決するために、この発
明にかかる酵素固定化電極は、導電性材料からなる電極
本体に酵素固定化膜および妨害除去膜が設けられている
ものにおいて、前記妨害除去股が保湿剤を含んでいるこ
どを特徴とする。Therefore, in order to solve the first problem described above, an enzyme-immobilized electrode according to the present invention is provided with an enzyme-immobilized membrane and an interference removal membrane on an electrode body made of a conductive material. Features a child whose crotch contains moisturizer.
上記第２の課題を解決するために、この発明にかかる酵
素固定化電極の製造方法は、導電性材料からなる電極本
体表面上に形成された妨害除去膜の上に酵素固定化膜を
形成するにあたり、前記妨害除去膜に保湿剤を含めてお
くことを特徴とずるこの発明の酵素固定化電極は、電極
本体表面上に少なくとも妨害除去膜および酵素固定化膜
を備えている。好ましくは、電極本体表面上に下地膜、
妨害除去膜および酵素固定化膜の３層を少なくとも備え
ている。In order to solve the above second problem, the method for manufacturing an enzyme-immobilized electrode according to the present invention includes forming an enzyme-immobilized film on an interference removal film formed on the surface of an electrode body made of a conductive material. The enzyme-immobilized electrode of the present invention is characterized in that the interference-removing film contains a humectant.The enzyme-immobilized electrode of the present invention is provided with at least an interference-removing film and an enzyme-immobilizing film on the surface of the electrode body. Preferably, a base film is provided on the surface of the electrode body,
It has at least three layers: an interference removal membrane and an enzyme immobilization membrane.
ここで、下地膜は、妨害除去膜の電極本体への密着性を
良くするために必要に応して用いられる。下地膜は、た
とえば、ゼラチン水溶液などの生体高分子水溶液に架橋
剤を添加したものを電極本体表面に塗布し、架橋反応を
行って形成された膜である。下地膜のマトリソクス戒分
はゼラチンに限定されない。架橋剤としては、たとえば
グルタルアルデヒド等のジアルデヒドが用いられる。Here, the base film is used as necessary to improve the adhesion of the interference removal film to the electrode body. The base film is a film formed by, for example, applying a crosslinking agent to an aqueous biopolymer solution such as an aqueous gelatin solution to the surface of the electrode body and performing a crosslinking reaction. The matrix content of the base film is not limited to gelatin. As the crosslinking agent, for example, a dialdehyde such as glutaraldehyde is used.
妨害除去膜は、マｌ・リソクス成分の少なくとも一部が
保湿剤であり、たとえば、アルブミン等のタンパク、ボ
リアリルアミン等の水溶性高分子、保湿剤および架橋剤
を含む溶液を電極本体または前記下地膜または酵素固定
化膜上に所定量を塗布し、架橋反応を行わせて、電極表
面上で製膜を行う。保湿剤としては、妨害除去膜の水分
を保ったりあるいは高めたりするものであれば特に限定
はないが、コラーゲン、ポリエチレングリコールアルキ
ル化合物などの界面活性剤、ポリビニルピロリドンやポ
リビニルアルコールなどの水溶性高分子があるが、妨害
除去膜の保湿性を保ち、かつ、妨害除去効果の低下が少
ないという点からは、コラーゲンを保湿剤として用いる
のが好ましい。コラーゲンとしては、酸処理コラーゲン
、アルカリ処理コラーゲン、酵素可熔性コラーゲンなど
のいずれも使用可能であり、コラーゲンのタイプも特に
限定はない。妨害除去膜を作製するための熔液（以下「
妨害除去膜作製液」と言う）では、コラーゲンの濃度は
、特に限定されないが、ｏ．ｏｉ〜０．１重量％の範囲
が好ましい。０．１重量％を越えると、膜の透過性が悪
くなり、応答速度が悪くなる。この発明にかかる酵素固
定化電極の応答性は、たとえば２０〜３０秒である。ま
た、妨害除去膜の架橋の程度によって除去されうる物質
の種類を適宜変えることも可能である。妨害除去膜のマ
トリックス戒分としては、保湿剤を含むのであれば、上
記のものに限定されない。架橋剤としては、たとえばグ
ルタルアルデヒド等のジアルデヒドが用いられる。At least a part of the mal-resox component is a humectant, and the interference removal film is a solution containing a protein such as albumin, a water-soluble polymer such as polyallylamine, a humectant, and a crosslinking agent, on the electrode body or under the electrode body. A predetermined amount of the solution is applied onto a geomembrane or an enzyme-immobilized membrane, a crosslinking reaction is performed, and a membrane is formed on the electrode surface. The moisturizer is not particularly limited as long as it retains or increases the moisture content of the interference removal membrane, but it may include collagen, surfactants such as polyethylene glycol alkyl compounds, and water-soluble polymers such as polyvinylpyrrolidone and polyvinyl alcohol. However, it is preferable to use collagen as a humectant in order to maintain the moisturizing properties of the interference removal film and to reduce the decrease in interference removal effect. As the collagen, any of acid-treated collagen, alkali-treated collagen, enzyme-fusible collagen, etc. can be used, and the type of collagen is not particularly limited. Melt solution (hereinafter referred to as “
In the "interference removal membrane preparation solution"), the concentration of collagen is not particularly limited, but the concentration of collagen is o. A range of oi to 0.1% by weight is preferred. If it exceeds 0.1% by weight, the permeability of the membrane will deteriorate and the response speed will deteriorate. The responsiveness of the enzyme-immobilized electrode according to the present invention is, for example, 20 to 30 seconds. Further, it is also possible to appropriately change the type of substance that can be removed depending on the degree of crosslinking of the interference removal membrane. The matrix components of the interference removal film are not limited to those mentioned above, as long as they include a humectant. As the crosslinking agent, for example, a dialdehyde such as glutaraldehyde is used.
酵素固定化膜は、グルコースオキシダーゼおよびコレス
テロールオキシダーゼ等の酵素、ゼラチン等の生体高分
子、および、架橋剤を含む溶液（たとえば、水溶液）を
電極本体上または妨害除去膜上に所定量塗布し、架橋反
応を行わせて製膜を行うとともに固定化して作られる。The enzyme-immobilized membrane is made by applying a predetermined amount of a solution (e.g., an aqueous solution) containing enzymes such as glucose oxidase and cholesterol oxidase, biopolymers such as gelatin, and a crosslinking agent onto the electrode body or onto the interference removal membrane, and then crosslinking the membrane. It is produced by conducting a reaction to form a film and immobilizing it.
酵素は、１種類のみを用いるようにしてもよいし、失活
などの悪影響が生しないのであれば、２種類以上を用い
るようにしてもよい。酵素固定化膜のマトリソクス威分
としては、酵素の働きを損なわないのであれば、上記の
ものに限定されない。架橋剤としては、たとえばグルタ
ルアルデヒド等のジアルデヒドが用いられる。Only one type of enzyme may be used, or two or more types may be used as long as no adverse effects such as deactivation occur. The matrix material for the enzyme-immobilized membrane is not limited to those mentioned above, as long as it does not impair the function of the enzyme. As the crosslinking agent, for example, a dialdehyde such as glutaraldehyde is used.
このようにして得られるこの発明にかかる酵素固定化電
極は、保湿剤によって妨害除去膜の水分が保たれるため
、保湿性を保つことができ、乾燥による妨害除去膜の電
極本体からの剥離を防くことができる。また、測定溶液
中での膨潤率、乾燥させたときの収縮率が小さくなり、
酵素固定化電極として使用したときの感度のハラッキが
なく、安定した測定を行うことができる。The enzyme-immobilized electrode according to the present invention obtained in this manner can maintain moisture retention because the moisture content of the interference removal film is retained by the humectant, and the separation of the interference removal film from the electrode body due to drying can be maintained. It can be prevented. In addition, the swelling rate in the measurement solution and the shrinkage rate when dried are reduced.
There is no fluctuation in sensitivity when used as an enzyme-immobilized electrode, and stable measurements can be performed.
また、妨害除去膜作製用の水溶液の塗布性も良くなり、
電極表面上に膜厚が均一になるように製膜することがで
きる。In addition, the applicability of the aqueous solution for preparing the interference removal membrane is improved,
The film can be formed to have a uniform thickness on the electrode surface.
保湿剤を含まない従来の妨害除去膜では、電極本体に妨
害除去膜作Ｍ肢を塗布後、調湿器の外、たとえば、湿度
４０〜６０％程度の雰囲気下に５〜１０分間放置すると
、第３図（ａｌにみるように、妨害除去膜５が電極本体
１から剥離してしまう。With conventional interference removal films that do not contain humectants, after applying the interference removal film to the electrode body, if you leave it outside a humidifier for 5 to 10 minutes in an atmosphere with a humidity of about 40 to 60%, As shown in FIG. 3 (al), the interference removal film 5 peels off from the electrode body 1.
このため、調湿器内で湿度８０％の雰囲気下に保って第
３図（ｂ）にみるように妨害除去欣４を作製ずる必要が
あった。これに対し、この発明にかかる酔素固定化電極
の製造方法では、妨害除去膜の」二に酵素固定化膜を形
成するにあたり、あらかしめ前記妨害除去膜に保湿剤を
含ませておくので、調湿器の外、たとえば、湿度４０〜
６０％程度の雰囲気下に５〜ＩＯ分間放置しても、第３
図ｔｂ＋にみるように妨害除去膜４が形成され、剥離が
生しにくい。第ｌ〜３図中、３は、セラミソク、合威樹
脂等からなる絶縁性基板である。電極本体１は、たとえ
ば、この基板３の上に形成されているが、このようなも
のに限定されない。For this reason, it was necessary to maintain the atmosphere at a humidity of 80% in a humidity conditioner and to manufacture the interference removal filter 4 as shown in FIG. 3(b). On the other hand, in the method for manufacturing an narcotic acid-immobilized electrode according to the present invention, before forming the enzyme-immobilized film on the second part of the interference-removing membrane, the interference-removing membrane is pre-impregnated with a humectant. Outside the humidifier, for example, humidity 40~
Even if left in an atmosphere of about 60% for 5 to IO minutes, the third
As shown in Figure tb+, the interference removal film 4 is formed and peeling is less likely to occur. In Figures 1 to 3, 3 is an insulating substrate made of ceramic material, Hei resin, or the like. The electrode body 1 is formed, for example, on this substrate 3, but is not limited thereto.
なお、この発明にかかる酵素固定化電極は、１０００回
以上の連続測定も可能である。Note that the enzyme-immobilized electrode according to the present invention is capable of continuous measurement of 1000 times or more.
坊害除去股が保湿剤を含んでいることにより、乾燥・湿
潤による収縮・膨潤の変化が小さくなり、剥離しにくく
なる。Since the hair removal crotch contains a humectant, changes in shrinkage and swelling due to dryness and moisture are reduced, making it difficult to peel off.
また、妨害除去膜が保湿剤を含んでいることにより、同
妨害除去膜の上に酵素固定化膜を形威するときの雰囲気
の調湿を行わずにすみ、妨害除去膜が電極本体から剥離
するのが防がれるという利点も得られる。In addition, since the interference removal film contains a humectant, there is no need to adjust the humidity of the atmosphere when forming the enzyme immobilization membrane on the interference removal film, and the interference removal film peels off from the electrode body. You also have the advantage of being prevented from doing so.
以下に、この発明の具体的な実施例および比較例を示す
が、この発明は下記実施例に限定されない。Specific examples and comparative examples of the present invention are shown below, but the present invention is not limited to the following examples.
実施例ｌ〜５および比較例
第１表にみるような組威の下地膜作製溶液、妨害除去膜
作製肢および酵素固定化ｆ６波を用いた。Examples 1 to 5 and Comparative Examples A base membrane preparation solution, an interference removing membrane preparation solution, and an enzyme-immobilized F6 wave of Kumihi as shown in Table 1 were used.
９
１０
妨害除去膜のマトリックス成分としてポリアリルア主ン
およびアルブミンを、架橋剤としてグルタルアルデヒド
を、保湿剤としてコラーゲンをそれぞれ用いた。酵素固
定化膜のマトリソクス威分は、ゼラチン、ポリアリルア
ミン、アルブミンであり、架橋剤はグルタルアルデヒド
である。下地＋１Ｑ作製液を第１図にみるように、リー
ド部２を有し、基板３上に設けられた電極本体（白金電
極）１表面上に所定量塗布したのら架椙反Ｌｌ＞を行わ
−Ｗて下地膜をｆｉ１膜した。この下地膜の表面に第１
表に示す組成の妨害除去膜作製液６を塗布して（第２図
参照）架橋反応を行わせ、妨害除去膜４を形威した（第
３図（ｂｌ参照）。この妨害除去膜４の表面に第１表に
示ず組威の酵素固定化溶液を塗布して架橋反応を行わせ
、酵素固定化膜を形成した。9 10 Polyallyl and albumin were used as matrix components of the interference removal membrane, glutaraldehyde was used as a crosslinking agent, and collagen was used as a humectant. The matrix components of the enzyme-immobilized membrane are gelatin, polyallylamine, and albumin, and the crosslinking agent is glutaraldehyde. As shown in Fig. 1, a predetermined amount of the base+1Q preparation solution is applied onto the surface of the electrode body (platinum electrode) 1, which has a lead part 2 and is provided on the substrate 3, and then a -W was used to form a fi1 base film. The first layer is applied to the surface of this base film.
The interference removal film preparation solution 6 having the composition shown in the table was applied (see Fig. 2) to cause a crosslinking reaction to form the interference removal film 4 (see Fig. 3 (bl). An enzyme immobilization solution prepared by Kumiui (not shown in Table 1) was applied to the surface to carry out a crosslinking reaction to form an enzyme immobilization membrane.
なお、比較例では、妨害除去映作製液を第２図にみるよ
うに塗布してから湿度４０〜６０％の雰囲気下に５〜１
０分間放置したところ、第３図（＝１）にみるように、
妨害除去膜５が電極本体１から剥離してしまった。この
ため、妨害除去映作製液を塗布して調湿器内に入れ、湿
度８０％以上の雰囲気下で妨害除去膜の作製（第３図（
ｂｌ参照）および酵素固定化膜の作製を行った。In addition, in the comparative example, after applying the interference removal film production liquid as shown in Figure 2, it was applied for 5 to 1 hour in an atmosphere with a humidity of 40 to 60%.
When left for 0 minutes, as shown in Figure 3 (=1),
The interference removal film 5 has peeled off from the electrode body 1. For this purpose, the interference removal film was prepared by applying the interference removal film production liquid and placing it in a humidity controller in an atmosphere with a humidity of 80% or more (see Figure 3).
bl) and an enzyme-immobilized membrane was prepared.
上記実施例および比較例で用いたボリアリルアミンは平
均分子量約６００００であり、実施例で用いたコラーゲ
ンは■高研製の牛真皮・可熔性タイフ゛Ｉコラーゲン冫
容液（ｐ Ｈ ３． ０、濃度０．３％）のものであっ
た。The polyallylamine used in the above Examples and Comparative Examples has an average molecular weight of about 60,000, and the collagen used in the Examples is Bovine Dermal/Flusible Type I Collagen Solution (pH 3.0, concentration 0.3%).
実施例１〜５および比較例の各酵素固定化電極について
検出感度（測定感度）、保湿性および感度変動を調べて
結果を第１表に示した。感度は、酵素固定化電極をポー
ラ口アナライザ等の測定装置に接続し、電圧０．６Ｖ（
対Ａｇ電極）、サンプル量を２０μＣとして測定した。The detection sensitivity (measurement sensitivity), moisture retention property, and sensitivity fluctuation were investigated for each enzyme-immobilized electrode of Examples 1 to 5 and Comparative Example, and the results are shown in Table 1. Sensitivity is determined by connecting the enzyme-immobilized electrode to a measuring device such as a polar analyzer, and applying a voltage of 0.6 V (
(Ag electrode), the sample amount was 20 μC.
サンプルは、１５０ｎｗ／ｄ１のグルコース溶液、１０
０■／ｄ１のコレステロール溶液、妨害物質として、ア
スコルビン酸（濃度２０■／ｄ１）、血岐の凝固防止な
どに使用されるＮａＦ（濃度５０■／ｄ！〉を用いた。The sample was a 150 nw/d1 glucose solution, 10
A cholesterol solution of 0 .mu./d1, ascorbic acid (concentration of 20 .mu./d1) and NaF (concentration of 50 .mu./d!), which is used to prevent coagulation of blood vessels, were used as interfering substances.
保湿性は、妨害除去膜作製液を塗布後、湿度４０〜６０
％の雰囲気下で５〜１０分間程度のうちにｌ１
１２
妨害除去膜が剥離しない場合を○、剥亜した場合を×で
示した。感度変動は、得られた酵素固定化電極を用いて
標準液で２０回測定したときの１回目の感度の測定値と
２０回目の感度の測定イ直との変化を下式により求めて
示した。Moisture retention is determined at a humidity of 40 to 60 after applying the interference removal film preparation solution.
A case in which the l1 12 interference removal film did not peel off within about 5 to 10 minutes in an atmosphere of 10% was indicated by ◯, and a case in which it did peel off was indicated by ×. Sensitivity fluctuation was determined by calculating the change between the first sensitivity measurement value and the 20th sensitivity measurement using the following formula when measuring 20 times with a standard solution using the obtained enzyme-immobilized electrode. .
１３
第１表からわかるように、実施例１〜３の酵素固定化電
極は、いずれもグルコースを感度良く検出することがで
き、実施例４および５の酵素固定化電極は、いずれもコ
レステロールを感度良く検出することができた。また、
実施例■〜５では、妨害物質はほとんどまたは全く検出
されなかった。これに対し、比較例の酵素固定化電極で
は、感度変動が実施例のもの（±２％以内）よりも大き
かった（±４〜５％）。また、比較例のものは、湿度４
０〜６０％ＲＨの雰囲気下では、第３図（ａｌにみるよ
うに、妨害除去膜５が電極本体１から剥離したが、実施
例のものでは、同し条件下で剥離しなかった。13 As can be seen from Table 1, the enzyme-immobilized electrodes of Examples 1 to 3 can all detect glucose with high sensitivity, and the enzyme-immobilized electrodes of Examples 4 and 5 can both detect cholesterol with high sensitivity. We were able to detect it well. Also,
In Examples 1-5, little or no interfering substances were detected. On the other hand, in the enzyme-immobilized electrode of the comparative example, the sensitivity fluctuation was larger (±4 to 5%) than that of the example (within ±2%). In addition, the comparative example has a humidity of 4
In an atmosphere of 0 to 60% RH, the interference removal film 5 peeled off from the electrode body 1 as shown in FIG. 3 (al), but in the example, it did not peel off under the same conditions.
なお、上記実施例では、固定化する酵素として、グルコ
ースオキシダーゼおよびコレステｒ：Ｉ−ルオキシダー
ゼを用いたが、この発明において使用できる酵素はこれ
らの２者に限定されるものではない。また、測定用の戒
も血渣に限るものではない。In the above example, glucose oxidase and cholesteryl oxidase were used as the enzymes to be immobilized, but the enzymes that can be used in the present invention are not limited to these two. Furthermore, the precepts for measurement are not limited to blood smear.
〔発リ１の効果〕
この発明にかかる酵素固定化電極は、導電性材料からな
る電極本体に酵素固定化膜および妨害除去股が設けられ
ているものにおいて、前記妨害除去膜が保湿剤を含んで
いることを特徴とするので、妨害除去膜の剥離がなく、
感度の良い、安定した測定を行うことができる。[Effect of First Effect] The enzyme-immobilized electrode according to the present invention has an enzyme-immobilized film and an interference removal crotch provided on an electrode body made of a conductive material, and the interference removal film does not contain a humectant. It is characterized by the fact that there is no peeling of the interference removal film,
Able to perform sensitive and stable measurements.
この発明にかかる酵素固定化電極の製造方法は、導電性
材料からなる電極本体表面上に形成された妨害除去膜の
上に酵素固定化膜を形成するにあたり、前記妨害除去映
に保湿剤を含めておくことを特徴とするので、妨害除去
膜の上に酵素固定化膜を形成するときに、調湿する必要
がない。In the method for producing an enzyme-immobilized electrode according to the present invention, when forming an enzyme-immobilized film on an interference-removing film formed on the surface of an electrode body made of a conductive material, a humectant is included in the interference-removing film. Therefore, there is no need to adjust the humidity when forming the enzyme-immobilized membrane on the interference removal membrane.
第１図は、実施例および比較例に用いた白金電極のｍ雌
を表す斜視図、第２図は、同白金電極に妨害除去膜作製
液を塗布した状態を表す概略断面図、第３図＋８＋は、
比較例の酵素固定化電極における妨害除去膜形威後で酵
素固定化膜形成前の状態を表す概略断面図、第３図ｆｂ
ｌは、実施例の酵素固定化電極における妨害除去膜形成
後で酵素固定化１５
１６
膜形威前の状態を表す概略断面図である。
１・・・電極本体 ４・・・妨害除去欣第１図FIG. 1 is a perspective view showing a female platinum electrode used in Examples and Comparative Examples, FIG. 2 is a schematic cross-sectional view showing the same platinum electrode coated with an interference removal film preparation solution, and FIG. +8+ is
Schematic cross-sectional view showing the state before formation of the enzyme-immobilized film after interference removal film formation in the enzyme-immobilized electrode of the comparative example, FIG. 3 fb
FIG. 1 is a schematic cross-sectional view showing the state in which the enzyme-immobilized 15 16 membrane is formed after the interference removal membrane is formed in the enzyme-immobilized electrode of the example. 1... Electrode body 4... Interference removal diagram Figure 1
Claims (1)
妨害物質除去膜が設けられている酵素固定化電極におい
て、前記妨害物質除去膜が保湿剤を含んでいることを特
徴とする酵素固定化電極。 ２ 導電性材料からなる電極本体表面上に形成された妨
害物質除去膜の上に酵素固定化膜を形成する酵素固定化
電極の製造方法において、前記妨害物質除去膜に保湿剤
を含めておくことを特徴とする酵素固定化電極の製造方
法。[Scope of Claims] 1. An enzyme-immobilized electrode in which an enzyme-immobilized film and an interfering substance removing film are provided on an electrode body made of a conductive material, characterized in that the interfering substance removing film contains a humectant. Enzyme-immobilized electrode. 2. In the method for producing an enzyme-immobilized electrode, in which an enzyme-immobilized film is formed on an interfering substance-removing film formed on the surface of an electrode body made of a conductive material, the interfering substance-removing film contains a humectant. A method for producing an enzyme-immobilized electrode characterized by:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1164797A JPH0750058B2 (en) | 1989-06-27 | 1989-06-27 | Enzyme-immobilized electrode and method for producing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1164797A JPH0750058B2 (en) | 1989-06-27 | 1989-06-27 | Enzyme-immobilized electrode and method for producing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0328752A true JPH0328752A (en) | 1991-02-06 |
| JPH0750058B2 JPH0750058B2 (en) | 1995-05-31 |
Family
ID=15800119
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1164797A Expired - Lifetime JPH0750058B2 (en) | 1989-06-27 | 1989-06-27 | Enzyme-immobilized electrode and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0750058B2 (en) |
Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06281614A (en) * | 1992-12-15 | 1994-10-07 | Avl Medical Instr Ag | Enzyme electrode for measuring current |
| US5741319A (en) * | 1995-01-27 | 1998-04-21 | Medtronic, Inc. | Biocompatible medical lead |
| US9610034B2 (en) | 2001-01-02 | 2017-04-04 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US9625413B2 (en) | 2006-03-31 | 2017-04-18 | Abbott Diabetes Care Inc. | Analyte monitoring devices and methods therefor |
| US9662057B2 (en) | 2000-06-27 | 2017-05-30 | Abbott Diabetes Care Inc. | Integrated sample acquisition and analyte measurement method |
| US9669162B2 (en) | 2005-11-04 | 2017-06-06 | Abbott Diabetes Care Inc. | Method and system for providing basal profile modification in analyte monitoring and management systems |
| US9668684B2 (en) | 2009-02-26 | 2017-06-06 | Abbott Diabetes Care Inc. | Self-powered analyte sensor |
| US9743863B2 (en) | 2006-03-31 | 2017-08-29 | Abbott Diabetes Care Inc. | Method and system for powering an electronic device |
| US9801545B2 (en) | 2007-03-01 | 2017-10-31 | Abbott Diabetes Care Inc. | Method and apparatus for providing rolling data in communication systems |
| US9891185B2 (en) | 1998-10-08 | 2018-02-13 | Abbott Diabetes Care Inc. | Small volume in vitro analyte sensor |
| US9962091B2 (en) | 2002-12-31 | 2018-05-08 | Abbott Diabetes Care Inc. | Continuous glucose monitoring system and methods of use |
| US9980670B2 (en) | 2002-11-05 | 2018-05-29 | Abbott Diabetes Care Inc. | Sensor inserter assembly |
| US10039881B2 (en) | 2002-12-31 | 2018-08-07 | Abbott Diabetes Care Inc. | Method and system for providing data communication in continuous glucose monitoring and management system |
| US10201301B2 (en) | 2005-11-01 | 2019-02-12 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US10231654B2 (en) | 2005-11-01 | 2019-03-19 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US10478108B2 (en) | 1998-04-30 | 2019-11-19 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| USD902408S1 (en) | 2003-11-05 | 2020-11-17 | Abbott Diabetes Care Inc. | Analyte sensor control unit |
| US11045147B2 (en) | 2009-08-31 | 2021-06-29 | Abbott Diabetes Care Inc. | Analyte signal processing device and methods |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63101743A (en) * | 1986-10-18 | 1988-05-06 | Matsushita Electric Works Ltd | Functional electrode |
-
1989
- 1989-06-27 JP JP1164797A patent/JPH0750058B2/en not_active Expired - Lifetime
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63101743A (en) * | 1986-10-18 | 1988-05-06 | Matsushita Electric Works Ltd | Functional electrode |
Cited By (31)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06281614A (en) * | 1992-12-15 | 1994-10-07 | Avl Medical Instr Ag | Enzyme electrode for measuring current |
| US5741319A (en) * | 1995-01-27 | 1998-04-21 | Medtronic, Inc. | Biocompatible medical lead |
| US10478108B2 (en) | 1998-04-30 | 2019-11-19 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US9891185B2 (en) | 1998-10-08 | 2018-02-13 | Abbott Diabetes Care Inc. | Small volume in vitro analyte sensor |
| US9662057B2 (en) | 2000-06-27 | 2017-05-30 | Abbott Diabetes Care Inc. | Integrated sample acquisition and analyte measurement method |
| US9610034B2 (en) | 2001-01-02 | 2017-04-04 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US11141084B2 (en) | 2002-11-05 | 2021-10-12 | Abbott Diabetes Care Inc. | Sensor inserter assembly |
| US10973443B2 (en) | 2002-11-05 | 2021-04-13 | Abbott Diabetes Care Inc. | Sensor inserter assembly |
| US9980670B2 (en) | 2002-11-05 | 2018-05-29 | Abbott Diabetes Care Inc. | Sensor inserter assembly |
| US11116430B2 (en) | 2002-11-05 | 2021-09-14 | Abbott Diabetes Care Inc. | Sensor inserter assembly |
| US9962091B2 (en) | 2002-12-31 | 2018-05-08 | Abbott Diabetes Care Inc. | Continuous glucose monitoring system and methods of use |
| US10039881B2 (en) | 2002-12-31 | 2018-08-07 | Abbott Diabetes Care Inc. | Method and system for providing data communication in continuous glucose monitoring and management system |
| US10750952B2 (en) | 2002-12-31 | 2020-08-25 | Abbott Diabetes Care 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 |
| USD902408S1 (en) | 2003-11-05 | 2020-11-17 | Abbott Diabetes Care Inc. | Analyte sensor control unit |
| US10231654B2 (en) | 2005-11-01 | 2019-03-19 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US11272867B2 (en) | 2005-11-01 | 2022-03-15 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US10201301B2 (en) | 2005-11-01 | 2019-02-12 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US11911151B1 (en) | 2005-11-01 | 2024-02-27 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US10952652B2 (en) | 2005-11-01 | 2021-03-23 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US11399748B2 (en) | 2005-11-01 | 2022-08-02 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US11363975B2 (en) | 2005-11-01 | 2022-06-21 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US11103165B2 (en) | 2005-11-01 | 2021-08-31 | Abbott Diabetes Care Inc. | Analyte monitoring device and methods of use |
| US11538580B2 (en) | 2005-11-04 | 2022-12-27 | Abbott Diabetes Care Inc. | Method and system for providing basal profile modification in analyte monitoring and management systems |
| US9669162B2 (en) | 2005-11-04 | 2017-06-06 | Abbott Diabetes Care Inc. | Method and system for providing basal profile modification in analyte monitoring and management systems |
| US9625413B2 (en) | 2006-03-31 | 2017-04-18 | Abbott Diabetes Care Inc. | Analyte monitoring devices and methods therefor |
| US9743863B2 (en) | 2006-03-31 | 2017-08-29 | Abbott Diabetes Care Inc. | Method and system for powering an electronic device |
| US9801545B2 (en) | 2007-03-01 | 2017-10-31 | Abbott Diabetes Care Inc. | Method and apparatus for providing rolling data in communication systems |
| US10631768B2 (en) | 2009-02-26 | 2020-04-28 | Abbott Diabetes Inc. | Self-powered analyte sensor |
| US9668684B2 (en) | 2009-02-26 | 2017-06-06 | Abbott Diabetes Care Inc. | Self-powered analyte sensor |
| US11045147B2 (en) | 2009-08-31 | 2021-06-29 | Abbott Diabetes Care Inc. | Analyte signal processing device and methods |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0750058B2 (en) | 1995-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JPH0328752A (en) | Enzyme immobilized electrode and its production | |
| US5082550A (en) | Enzyme electrochemical sensor electrode and method of making it | |
| US5284140A (en) | Acrylic copolymer membranes for biosensors | |
| CA2394948C (en) | Screen-printable paste for producing a porous polymer membrane for a biosensor | |
| EP0079502B1 (en) | Multilayer enzyme electrode membrane, method of making same and polarographic cell structure | |
| JP3950174B2 (en) | Glucose sensor | |
| US4240889A (en) | Enzyme electrode provided with immobilized enzyme membrane | |
| JP4055912B2 (en) | Glucose sensor | |
| JPS5886450A (en) | Enzyme electrode film and its manufacture | |
| JPH02211167A (en) | Supplying method for capsula containing hydragel of polyvinyl base and biochemically active material,on base material | |
| JPH0235933B2 (en) | ||
| GB2209836A (en) | Multilayer enzyme electrode membrane and method of making same | |
| JP3447374B2 (en) | Enzyme sensor and method for producing the same | |
| JPS6264941A (en) | Enzyme immobilized membrane for biosensor | |
| JP2814027B2 (en) | Biosensor | |
| JP3064599B2 (en) | Glucose biosensor | |
| JPS5896246A (en) | Enzyme electrode membrane, enzyme thereof is protected and wrapped by capsule, and its manufacture | |
| JPH0222552A (en) | Stylus electrode | |
| JP3064635B2 (en) | Glucose biosensor | |
| JPH0315692B2 (en) | ||
| JPS59228160A (en) | Amylase analyzing method | |
| JPH0599882A (en) | Glucose bio-sensor | |
| JPH03277956A (en) | Enzyme immobilized electrode device | |
| Gregory et al. | Fabrication methods for integrated biosensors | |
| JPH0723882B2 (en) | Biosensor and sensor plate |