JPH0481740B2 - - Google Patents
Info
- Publication number
- JPH0481740B2 JPH0481740B2 JP61070150A JP7015086A JPH0481740B2 JP H0481740 B2 JPH0481740 B2 JP H0481740B2 JP 61070150 A JP61070150 A JP 61070150A JP 7015086 A JP7015086 A JP 7015086A JP H0481740 B2 JPH0481740 B2 JP H0481740B2
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- immobilized
- film
- photoresist
- isfet
- 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.)
- Expired - Lifetime
Links
- 229920002120 photoresistant polymer Polymers 0.000 claims description 34
- 239000012528 membrane Substances 0.000 claims description 28
- 239000004065 semiconductor Substances 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 22
- 230000005669 field effect Effects 0.000 claims description 12
- 150000002500 ions Chemical class 0.000 description 10
- 108090000790 Enzymes Proteins 0.000 description 9
- 102000004190 Enzymes Human genes 0.000 description 9
- 229940088598 enzyme Drugs 0.000 description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical group CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 6
- 229910052710 silicon Inorganic materials 0.000 description 6
- 239000010703 silicon Substances 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- 239000003960 organic solvent Substances 0.000 description 5
- 229910052594 sapphire Inorganic materials 0.000 description 5
- 239000010980 sapphire Substances 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 4
- 239000003431 cross linking reagent Substances 0.000 description 4
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 4
- 239000010931 gold Substances 0.000 description 4
- 229910052737 gold Inorganic materials 0.000 description 4
- 238000000206 photolithography Methods 0.000 description 4
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000012460 protein solution Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- UMVBXBACMIOFDO-UHFFFAOYSA-N [N].[Si] Chemical compound [N].[Si] UMVBXBACMIOFDO-UHFFFAOYSA-N 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000002093 peripheral effect Effects 0.000 description 2
- 229910052814 silicon oxide Inorganic materials 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000004528 spin coating Methods 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 229920002284 Cellulose triacetate Polymers 0.000 description 1
- 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 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- NNLVGZFZQQXQNW-ADJNRHBOSA-N [(2r,3r,4s,5r,6s)-4,5-diacetyloxy-3-[(2s,3r,4s,5r,6r)-3,4,5-triacetyloxy-6-(acetyloxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6s)-4,5,6-triacetyloxy-2-(acetyloxymethyl)oxan-3-yl]oxyoxan-2-yl]methyl acetate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](OC(C)=O)[C@H]1OC(C)=O)O[C@H]1[C@@H]([C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H](COC(C)=O)O1)OC(C)=O)COC(=O)C)[C@@H]1[C@@H](COC(C)=O)O[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@H]1OC(C)=O NNLVGZFZQQXQNW-ADJNRHBOSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-N ethanesulfonic acid Chemical compound CCS(O)(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-N 0.000 description 1
- MUJOIMFVNIBMKC-UHFFFAOYSA-N fludioxonil Chemical compound C=12OC(F)(F)OC2=CC=CC=1C1=CNC=C1C#N MUJOIMFVNIBMKC-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は半導体電界効果型イオンセンサの表面
に酵素固定化膜が設けられてなる集積化された半
導体バイオセンサにおける酵素固定化膜の形成方
法に関するものである。Detailed Description of the Invention (Field of Industrial Application) The present invention relates to a method for forming an enzyme-immobilized membrane in an integrated semiconductor biosensor in which an enzyme-immobilized membrane is provided on the surface of a semiconductor field-effect ion sensor. It is related to.
（従来の技術）
従来、溶液中の特定の有機物の濃度を測定する
半導体バイオセンサーの一種に半導体電界酵素固
定化膜効果型イオンセンサ（以下ISFET）の表
面に酵素を固定化した膜が設けられたものが知ら
れている。このバイオセンサは、溶液中の特定の
有機物が酵素固定化膜中で酵素の触媒作用により
化学反応をした時に生じる水素イオン濃度の変化
をISFETで検出することにより、特定の有機物
の濃度を測定するものである。この選択性をもつ
酵素固定化膜の例として、尿素検出用としてウレ
アーゼ固定化酵素固定化膜、グルコース検出用と
してグルコースオキシダーゼ膜等が知られている
（センサーズ アンド アクチユエイターズ
（Sensors and Actuators）第７巻１頁〜10頁
（1985））。また、サフアイア基板上に設けられた
島状シリコンを用いて酵素固定化膜が設けられた
ISFETと失活した酵素固定化膜が設けられた
ISFETを同一チツプ上に形成し、裏面に参照電
局として金電極を設けることにより、ワンチツプ
化されたバイオセンサも開発されている（第16
回、1984インタナシヨナル カンフアレンス オ
ン ソリツド ステート デバイシズ アンド
マテイアルズ、レイト ニユーズ アブストラク
ツ（1984 International Conference on Solid
State Devices and Materi−als，Late News
Abstracts）66頁〜67頁（1984））。このようなバ
イオセンサを製造するにあたり、所定のISFET
上に酵素固定化膜を形成する必要がある。酸素固
定化膜の形成方法にはいくつかの方法が知られて
いるが、ウエハの段階で酵素固定化膜を形成でき
バイオセンサの大量生産を可能ならしめるものと
しては、例えば前記引用文献のように三酢酸セル
ロースを担体とした酵素固定化膜をウエハ全面に
形成した後、フオトマスクを介して紫外線を照
射、所定のISFET上以外の酵素固定化膜中の酵
素を失活させるという方法がある。また、フオト
レジストを塗布したウエハ上に酵素固定化膜を形
成した後に、所定のISFET上以外の酵素固定化
膜をリフトオフ法により除去するという方法も提
案されている（特願昭59−209165）。(Prior technology) Conventionally, a type of semiconductor biosensor that measures the concentration of a specific organic substance in a solution is a semiconductor electrolytic enzyme-immobilized membrane effect ion sensor (hereinafter referred to as ISFET), which has a membrane on which an enzyme is immobilized on the surface. things are known. This biosensor measures the concentration of specific organic substances by using ISFET to detect changes in hydrogen ion concentration that occur when specific organic substances in a solution undergo a chemical reaction due to the catalytic action of an enzyme in an enzyme-immobilized membrane. It is something. Examples of enzyme-immobilized membranes with this selectivity include urease-immobilized enzyme-immobilized membranes for urea detection and glucose oxidase membranes for glucose detection (Sensors and Actuators). ) Vol. 7, pp. 1-10 (1985)). In addition, an enzyme immobilization membrane was provided using island-like silicon provided on a saphire substrate.
An ISFET and a deactivated enzyme-immobilized membrane were provided.
A one-chip biosensor has also been developed by forming ISFETs on the same chip and providing a gold electrode as a reference voltage on the back side (No. 16).
1984 International Conference on Solid State Devices and
Materials, Late News Abstracts (1984 International Conference on Solid
State Devices and Materials, Late News
Abstracts) pp. 66-67 (1984)). In manufacturing such a biosensor, a certain ISFET
It is necessary to form an enzyme-immobilized membrane on top. Several methods are known for forming an oxygen-immobilized film, but the method described in the cited document above is one that allows the formation of an enzyme-immobilized film at the wafer stage and enables mass production of biosensors. Another method is to form an enzyme-immobilized film using cellulose triacetate as a carrier over the entire surface of the wafer, and then irradiate it with ultraviolet rays through a photomask to deactivate the enzymes in the enzyme-immobilized film except on the designated ISFET. A method has also been proposed in which, after forming an enzyme-immobilized film on a wafer coated with a photoresist, the enzyme-immobilized film other than those on designated ISFETs is removed by a lift-off method (Japanese Patent Application No. 59-209165). .
（発明が解決しようとする問題点）
上記リフトオフ法は短時間のうちに工程を終了
出来るという長所を有するが、形成された酵素固
定化膜の周辺部が著しく厚くなることがあつた。
バイオセンサの特性は酵素固定化膜の中央部の厚
さによつて決定されるため、ほとんどの場合この
ような厚みの増大はバイオセンサの製造上問題と
ならないが、肥厚化した部分の一部がバイオセン
サを測定に供した際に脱落することがあり、被測
定試料を汚染することが希にあるという問題点が
あつた。(Problems to be Solved by the Invention) The above-mentioned lift-off method has the advantage of being able to complete the process in a short period of time, but the peripheral portion of the formed enzyme-immobilized membrane may become extremely thick.
Since the characteristics of a biosensor are determined by the thickness of the central part of the enzyme-immobilized membrane, in most cases this increase in thickness does not pose a problem in the production of biosensors. There was a problem in that when the biosensor was used for measurement, it sometimes fell off, and in rare cases it could contaminate the sample to be measured.
（問題点を解決するための手段）
本発明は前記問題点を解決する手段として、１
つのチツプに１つまたは２つ以上の半導体電界効
果型イオンセンサが集積化され、そのうちの少な
くとも１つの半導体電界効果型イオンセンサの表
面に酵素固定化膜が設けられてなる半導体バイオ
センサの製造方法において、
(イ) 半導体電界効果型イオンセンサが形成された
半導体ウエハ上に有機溶剤に可溶なフオトレジ
ストを塗布した後、フオトリンググラフイ法に
より酵素固定化膜が設けられるべき所定の半導
体電界効果型イオンセンサの表面のフオトレジ
ストを除く工程と、
(ロ) 前記半導体ウエハ表面に、親水性プライマ溶
液をスピン塗布し、前記所定の半導体電界効果
型イオンセンサの表面を親水性プライマ処理す
る工程と、
(ハ) 前記工程を経た半導体表面に酵素と架橋剤を
含む蛋白質溶液をスピン塗布して前記半導体ウ
エハ表面上に酵素固定化膜を形成する工程と、
(ニ) さらに前記工程を経た半導体ウエハを前記フ
オトレジストを溶解する有機溶剤で処理して前
記フオトレジストを溶解し、前記所定の半導体
電界効果型イオンセンサの表面以外に存在する
酵素固定化膜をリフトオフによる除去する工程
とを備え前記フオトリソグラフイ法により酵素
固定化膜が設けられるべき所定の半導体電界効
果型イオンセンサの表面のフオトレジストを除
く工程で、フオトレジストが除かれた半導体電
界効果型イオンセンサノ表面を囲むフオトレジ
スト層の端面が前記半導体電界効果型イオンセ
ンサの表面に対してなだらかな傾斜をなすよう
にすることを特徴とする半導体バイオセンサ酵
素固定化膜の形成方法を提供するものである。(Means for solving the problems) The present invention provides means for solving the problems as follows:
A method for producing a semiconductor biosensor, in which one or more semiconductor field-effect ion sensors are integrated on a single chip, and an enzyme-immobilized membrane is provided on the surface of at least one of the semiconductor field-effect ion sensors. (a) After coating a photoresist soluble in an organic solvent on a semiconductor wafer on which a semiconductor field-effect ion sensor is formed, a predetermined semiconductor electric field is applied using a photoringography method to provide an enzyme-immobilized film. (b) Spin-coating a hydrophilic primer solution on the surface of the semiconductor wafer to treat the surface of the predetermined semiconductor field-effect ion sensor with a hydrophilic primer. (c) forming an enzyme-immobilized film on the semiconductor wafer surface by spin-coating a protein solution containing an enzyme and a crosslinking agent on the semiconductor surface that has undergone the above process; and (d) the semiconductor that has also undergone the above process. treating the wafer with an organic solvent that dissolves the photoresist to dissolve the photoresist, and removing the enzyme-immobilized film present on a surface other than the surface of the predetermined semiconductor field effect ion sensor by lift-off. A photoresist layer surrounding the surface of a semiconductor field-effect ion sensor from which the photoresist has been removed in a step of removing the photoresist on the surface of a given semiconductor field-effect ion sensor on which an enzyme-immobilized membrane is to be provided by photolithography. The present invention provides a method for forming an enzyme-immobilized membrane for a semiconductor biosensor, characterized in that the end face of the membrane is gently sloped with respect to the surface of the semiconductor field-effect ion sensor.
（作用）
本発明の酵素固定化膜の形成方法によれば、半
導体ウエハ上に有機溶剤に可溶なフオトレジスト
を塗布した後、フオトリソグラフイ法により酵素
固定化膜が設けられるべき所定のISFET上の表
面のフオトジストを除き、その上から親水性プラ
イマをスピン塗布する。フオトレジストが除かれ
たISFETの表面はこの工程により親水性プライ
マ処理され、後に形成される酵素固定化膜が
ISFETの表面より剥離することを防止する。こ
の上に酵素と架橋剤を含む蛋白質溶液を塗布する
わけであるが、フオトリソグラフイ法によつてフ
オトレジストが除かれたISFETの表面にはこれ
と接して酵素固定化膜が形成され、それ以外の部
分ではウエハ表面と形成された酵素固定化膜との
間にフオトレジスト層が存在するわけである。架
橋剤による架橋反応が終了するのをまつてから半
導体ウエハを有機溶剤で処理すると、フオトレジ
ストは有機溶剤に溶解し、その際フオトレジスト
上に形成された酵素固定化膜も剥離する。その結
果、ISFET表面に形成された酵素固定化膜のみ
が残る。以上の工程においてフオトリソグラフイ
法によつて所定のISFET表面上のフオトレジス
トを除いた際、ISFETの表面を囲むフオトレジ
スト４３の端面４１が第４図のようにISFET表
面４２に対し直角をなしていると、第５図に示し
たように形成された酵素固定化膜５１の形状はそ
の周辺部で著しく厚くなつた。本発明の方法によ
れば、ISFETの表面を囲むフオトレジストの端
面が第１図１１のようにISFETの表面１２に対
してなだらかな傾斜をなすようにすることによ
り、リフトオフの工程を経て形成される酵素固定
化膜の形状を第２図２１にほぼ平坦にすることが
可能となつた。(Function) According to the method for forming an enzyme-immobilized film of the present invention, a photoresist soluble in an organic solvent is applied onto a semiconductor wafer, and then a predetermined ISFET on which an enzyme-immobilized film is to be provided is formed by photolithography. Remove the top surface photogist and spin coat a hydrophilic primer over it. The surface of the ISFET from which the photoresist has been removed is treated with a hydrophilic primer in this step, and the enzyme-immobilized film formed later is coated with a hydrophilic primer.
Prevents peeling from the surface of ISFET. A protein solution containing an enzyme and a cross-linking agent is applied onto this, and an enzyme-immobilized film is formed in contact with the surface of the ISFET from which the photoresist has been removed by photolithography. In other parts, a photoresist layer exists between the wafer surface and the formed enzyme immobilization film. If the semiconductor wafer is treated with an organic solvent after waiting for the completion of the crosslinking reaction by the crosslinking agent, the photoresist will be dissolved in the organic solvent, and at this time, the enzyme-immobilized film formed on the photoresist will also be peeled off. As a result, only the enzyme-immobilized film formed on the ISFET surface remains. When the photoresist on a predetermined ISFET surface is removed by photolithography in the above process, the end face 41 of the photoresist 43 surrounding the ISFET surface is perpendicular to the ISFET surface 42 as shown in FIG. As a result, the shape of the enzyme-immobilized membrane 51 formed as shown in FIG. 5 became significantly thicker at its periphery. According to the method of the present invention, the end face of the photoresist surrounding the surface of the ISFET forms a gentle slope with respect to the surface 12 of the ISFET as shown in FIG. It became possible to make the shape of the enzyme-immobilized membrane almost flat as shown in FIG. 21.
（実施例）
以下本発明の実施例について図面を参照して説
明する。(Example) Examples of the present invention will be described below with reference to the drawings.
第１図は本発明による半導体バイオセンサ酵素
固定化膜の形成方法の一実施例の工程説明図で、
サフアイア基板上に形成された２つ一組の
ISFETの一方にのみ酵素固定化膜を形成する場
合について示してある。第３図ａ〜ｄにおいて、
３１はサフアイア基板、３２は高不純物濃度ｎ形
シリコン領域、３３はｐ形シリコン領域、３４は
酸化シリコン膜、３５は窒素シリコン膜、３６は
有機溶可溶性のフオトレジスト層、３７は金電極
である。次に製造工程を順を追つて説明する。サ
フアイア基板表面の島状シリコン層を用いて
ISFETを形成し、サフアイア基板裏面金を蒸着
したウエハ表面にフオトレジスト層、例えばシツ
プレー社製マイクロポジツト1300−37をスピン塗
布する（第３図ａ）。このフオトレジストはアセ
トン可溶性である。次に、フオトマスクを用いで
露光を行う。このとき焼付け時のプロキシミテイ
ーギヤツプ量を20μｍに設定する。現像により酵
素固定化膜が設けられるISFETの表面のフオト
レジスト層を除去する（第３図ｂ）。以上の工程
によりフオトレジストが除かれたISFETの表面
に対しその周辺を囲むフオトレジスト層の端面が
なだらかな傾斜をなすようにすることができる。
その後、親水性プライマ、例えばγ−アミノプロ
ピルトリエトキシシランの１％水溶液をウエハ上
にスピン塗布し、110℃で５分間熱処理を行い、
酵素固定化膜が設けられるISFETの表面にγ−
アミノプロピルトリエトキシシランを結合させ
る。このウエハを５％グルタルアルデヒド水溶液
に15分間浸漬した後、水洗、乾燥してから、次に
酸素と架橋剤を含む蛋白質溶液、例えば尿素を検
出する場合には300mg／ml牛血清アルブミンを含
む0.1Mピペラジン−Ｎ，N′−ビス（２−エタン
スルフオン酸）−水酸化ナトリウム（PH6.8）２体
積部に50mg／mlのウレアーゼ（マイルスラボラト
リーズ製53u／mg）水溶液１体積部を加え、さら
に2wt％グルタルアルデヒド水溶液１体積部を加
えた後よく混合した溶液0.9mlをスピン塗付する。
30分常温で放置してグルタルアルデヒドによる架
橋反応を完了させ酵素固定化膜３８を形成する。
このようにして酵素固定化膜が形成されたウエハ
（第３図ｃ）をアセトン中に浸漬し超音波洗浄器
を用いて２分間超音波処理を行うと、フオトレジ
スト層はアセトン中に溶出し、それとともフオト
レジスト層上に形成された酵素固定化膜を剥離す
る。その結果、あらかじめフオトレジスト膜が除
去されてあつたISFET上に形成された酵素固定
化膜３８のみがウエハ上に残る（第１図ｄ）。以
上の工程により所定のISFETの表面にだけに0.5μ
ｍの均一な厚さの酵素固定化膜を形成することが
できた。 FIG. 1 is a process explanatory diagram of an embodiment of the method for forming a semiconductor biosensor enzyme-immobilized membrane according to the present invention.
A set of two formed on a sapphire substrate.
A case is shown in which an enzyme-immobilized membrane is formed only on one side of the ISFET. In Figures 3a-d,
31 is a sapphire substrate, 32 is a high impurity concentration n-type silicon region, 33 is a p-type silicon region, 34 is a silicon oxide film, 35 is a nitrogen silicon film, 36 is an organic soluble photoresist layer, and 37 is a gold electrode. . Next, the manufacturing process will be explained step by step. Using an island-like silicon layer on the surface of a sapphire substrate
A photoresist layer, such as Microposit 1300-37 manufactured by Shipley Co., Ltd., is spin-coated on the surface of the wafer on which ISFETs are formed and gold is deposited on the back side of the sapphire substrate (FIG. 3a). This photoresist is acetone soluble. Next, exposure is performed using a photomask. At this time, the proximity gap amount during baking is set to 20 μm. The photoresist layer on the surface of the ISFET on which the enzyme immobilization film is provided is removed by development (FIG. 3b). Through the above steps, the end face of the photoresist layer surrounding the surface of the ISFET from which the photoresist has been removed can be made to form a gentle slope.
Thereafter, a hydrophilic primer, such as a 1% aqueous solution of γ-aminopropyltriethoxysilane, is spin-coated onto the wafer, and heat treated at 110°C for 5 minutes.
γ-
Attach aminopropyltriethoxysilane. The wafer is immersed in a 5% glutaraldehyde aqueous solution for 15 minutes, washed with water, dried, and then processed with a protein solution containing oxygen and a cross-linking agent, such as 0.1 containing 300 mg/ml bovine serum albumin in the case of detecting urea. To 2 parts by volume of M-piperazine-N,N'-bis(2-ethanesulfonic acid)-sodium hydroxide (PH6.8) was added 1 part by volume of a 50 mg/ml urease (manufactured by Miles Laboratories, 53 u/mg) aqueous solution, and an additional 2 wt. After adding 1 part by volume of % glutaraldehyde aqueous solution, spin-apply 0.9 ml of the well-mixed solution.
The crosslinking reaction by glutaraldehyde is completed by leaving it at room temperature for 30 minutes, and the enzyme-immobilized membrane 38 is formed.
When the wafer (Fig. 3c) on which the enzyme-immobilized film is formed in this way is immersed in acetone and subjected to ultrasonic treatment for 2 minutes using an ultrasonic cleaner, the photoresist layer is eluted into the acetone. At the same time, the enzyme-immobilized film formed on the photoresist layer is peeled off. As a result, only the enzyme-immobilized film 38 formed on the ISFET from which the photoresist film has been removed remains on the wafer (FIG. 1d). Through the above process, 0.5μ is applied only to the surface of the specified ISFET.
It was possible to form an enzyme-immobilized membrane with a uniform thickness of m.
（発明の効果）
本発明の方法によれば、酵素固定化膜はその周
辺部で肥厚化することなく、均一な厚さをもつて
所定のISFET上にのみ形成される。(Effects of the Invention) According to the method of the present invention, the enzyme-immobilized membrane is formed only on a predetermined ISFET with a uniform thickness without thickening in the peripheral area.
第１図は本発明にかかるフオトレジスト層の断
面図、第２図は本発明の方法によつて形成された
酵素固定化膜の断面図、第３図ａ〜ｄは本発明の
一実施例の工程説明図、第４図は従来法によるフ
オトレジスト層の断面図、第５図は従来法によつ
て形成された酵素固定化膜の断面図。
図において、１１はフオトレジストの端面、１
２はISFETの表面、１３はフオトレジスト層、
２１は酵素固定化膜、２２はISFETの表面、３
１はサフアイア基板、３２は高不純物濃度ｎ形シ
リコン領域、３３はｐ形シリコン領域、３４は酸
化シリコン膜、３５は窒素シリコン膜、３６はフ
オトレジスト膜、３７は金電極、３８は酵素固定
化膜、４１はフオトレジストの端面、４２は
ISFETの表面、４３はフオトレジスト層、５１
は酵素固定化膜で示す。
FIG. 1 is a cross-sectional view of a photoresist layer according to the present invention, FIG. 2 is a cross-sectional view of an enzyme-immobilized membrane formed by the method of the present invention, and FIGS. 3 a to 3 d are examples of the present invention. FIG. 4 is a cross-sectional view of a photoresist layer formed by a conventional method, and FIG. 5 is a cross-sectional view of an enzyme-immobilized film formed by a conventional method. In the figure, 11 is the end face of the photoresist, 1
2 is the surface of ISFET, 13 is the photoresist layer,
21 is the enzyme-immobilized membrane, 22 is the surface of ISFET, 3
1 is a sapphire substrate, 32 is a high impurity concentration n-type silicon region, 33 is a p-type silicon region, 34 is a silicon oxide film, 35 is a nitrogen silicon film, 36 is a photoresist film, 37 is a gold electrode, and 38 is an enzyme immobilization 41 is the end face of the photoresist; 42 is the film;
ISFET surface, 43 is photoresist layer, 51
is shown as an enzyme-immobilized membrane.
Claims (1)
固定化膜が設けられてなる半導体バイオセンサの
酵素固定化膜の形成方法において、フオトレジス
トを用いてリフトオフにより酵素固定化膜を形成
する際に、酵素固定化膜が設けられるべき所定の
半導体電界効果型イオンセンサの表面を囲むフオ
トレジスト層の端面を前記半導体電解効果型イオ
ンセンサの表面に対して傾斜をなすように形成す
ることを特徴とする半導体バイオセンサ酵素固定
化膜の形成方法。1. In a method for forming an enzyme-immobilized film for a semiconductor biosensor in which an enzyme-immobilized film is provided on the surface of a semiconductor field-effect ion sensor, when forming the enzyme-immobilized film by lift-off using a photoresist, the enzyme-immobilized film is A semiconductor characterized in that an end face of a photoresist layer surrounding a surface of a predetermined semiconductor field-effect ion sensor on which an immobilization film is to be provided is formed so as to be inclined with respect to the surface of the semiconductor field-effect ion sensor. Method for forming biosensor enzyme-immobilized membrane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61070150A JPS62225942A (en) | 1986-03-27 | 1986-03-27 | Method of forming semiconductor biosensor enzyme immobilized membrane |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61070150A JPS62225942A (en) | 1986-03-27 | 1986-03-27 | Method of forming semiconductor biosensor enzyme immobilized membrane |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS62225942A JPS62225942A (en) | 1987-10-03 |
| JPH0481740B2 true JPH0481740B2 (en) | 1992-12-24 |
Family
ID=13423262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61070150A Granted JPS62225942A (en) | 1986-03-27 | 1986-03-27 | Method of forming semiconductor biosensor enzyme immobilized membrane |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62225942A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05281181A (en) * | 1992-03-30 | 1993-10-29 | Nippon Telegr & Teleph Corp <Ntt> | Enzyme modified electrochemical detector and its manufacture |
-
1986
- 1986-03-27 JP JP61070150A patent/JPS62225942A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS62225942A (en) | 1987-10-03 |
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