JPS62225942A - Method of forming semiconductor biosensor enzyme immobilized membrane - Google Patents
Method of forming semiconductor biosensor enzyme immobilized membraneInfo
- Publication number
- JPS62225942A JPS62225942A JP61070150A JP7015086A JPS62225942A JP S62225942 A JPS62225942 A JP S62225942A JP 61070150 A JP61070150 A JP 61070150A JP 7015086 A JP7015086 A JP 7015086A JP S62225942 A JPS62225942 A JP S62225942A
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- membrane
- immobilized
- photoresist
- film
- 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
- 239000012528 membrane Substances 0.000 title claims abstract description 30
- 239000004065 semiconductor Substances 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims description 23
- 108090000790 Enzymes Proteins 0.000 title abstract description 15
- 102000004190 Enzymes Human genes 0.000 title abstract description 15
- 229920002120 photoresistant polymer Polymers 0.000 claims abstract description 33
- 230000005669 field effect Effects 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 3
- 229910052594 sapphire Inorganic materials 0.000 abstract description 7
- 239000010980 sapphire Substances 0.000 abstract description 7
- 239000000758 substrate Substances 0.000 abstract description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 abstract description 6
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 abstract description 6
- 229910052710 silicon Inorganic materials 0.000 abstract description 6
- 239000010703 silicon Substances 0.000 abstract description 6
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 abstract description 5
- 239000010931 gold Substances 0.000 abstract description 5
- 229910052737 gold Inorganic materials 0.000 abstract description 5
- 239000003431 cross linking reagent Substances 0.000 abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical group CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 abstract description 2
- 239000012460 protein solution Substances 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 10
- 150000002500 ions Chemical class 0.000 description 9
- 239000000243 solution Substances 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 5
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 238000000206 photolithography Methods 0.000 description 3
- 210000004885 white matter Anatomy 0.000 description 3
- 229910052581 Si3N4 Inorganic materials 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 238000010586 diagram 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
- HQVNEWCFYHHQES-UHFFFAOYSA-N silicon nitride Chemical compound N12[Si]34N5[Si]62N3[Si]51N64 HQVNEWCFYHHQES-UHFFFAOYSA-N 0.000 description 2
- 229910052814 silicon oxide Inorganic materials 0.000 description 2
- IHPYMWDTONKSCO-UHFFFAOYSA-N 2,2'-piperazine-1,4-diylbisethanesulfonic acid Chemical compound OS(=O)(=O)CCN1CCN(CCS(O)(=O)=O)CC1 IHPYMWDTONKSCO-UHFFFAOYSA-N 0.000 description 1
- WIGIZIANZCJQQY-UHFFFAOYSA-N 4-ethyl-3-methyl-N-[2-[4-[[[(4-methylcyclohexyl)amino]-oxomethyl]sulfamoyl]phenyl]ethyl]-5-oxo-2H-pyrrole-1-carboxamide Chemical compound O=C1C(CC)=C(C)CN1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)NC2CCC(C)CC2)C=C1 WIGIZIANZCJQQY-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 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
- 235000016496 Panda oleosa Nutrition 0.000 description 1
- 240000000220 Panda oleosa Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 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
- 239000013040 bath agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000004202 carbamide Substances 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
- 238000001035 drying Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 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
- 238000010438 heat treatment Methods 0.000 description 1
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 description 1
- 238000001459 lithography Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000002500 microbody Anatomy 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- KKCBUQHMOMHUOY-UHFFFAOYSA-N sodium oxide Chemical compound [O-2].[Na+].[Na+] KKCBUQHMOMHUOY-UHFFFAOYSA-N 0.000 description 1
- 229910001948 sodium oxide Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000004528 spin coating Methods 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
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
イオセンサの一種に半導体′１界ｅＮ素固定化膜効果型
イオンセンナ（以下１８ＦＪＧＴ）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＃度の変化をｌ５ＦＥＴ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 アクテユエイターズ（，８ｅｎｓｏｒｓ ａｎｄ
Ａｃｔｕａｔｏｒｓ ）第７巻１頁〜１０頁（１９８
５））。また、サファイア基板上に設けられた島状シリ
コンを周込て酵素固定化膜が設けられたｌ８ＦＥＴ（！
＝失活した酵素固定化膜が設けられたｌ５ＦＥＴを同一
チップ上に形成し、裏面に参照１局として金電極を設け
ることにより、フンチッ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
ンンアレンスオン ソリッド ステート デバイシズ
アンドマテリアルズ、レイト ニーーズ アプストラク
ツ （１９８４１ｎｔｅｒｎａｔｉｏｎａｌ Ｃｏ
ｎｆｅｒｅｎｃｅｏｎ ５ｏｌｉｄ ５ｔａｔｅ Ｄｅ
ｖｉｃｅｓ ａｎｄ Ｍａｔｅｒｉ −ａｌｓ、 Ｌ
ａｔｅ 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ＩＳＦｇＴ上に酵素固定イ瞑を
形成する必要がある。酵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、ウェ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ェハ全面に形成
（−た後、フォトマスクを介して紫外線を照射、所定の
１８ＦＥＴ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、フォトレジストを塗布したウ
ェハ上に酵素固定化膜を形成した後に、所定のｌ５ＦＥ
Ｔ上以外の酵素固定化膜をリフトオフ法により除去する
と因り方法も提案されている（特願昭５９−２０９１６
５ ）。(Prior art) Conventionally, as a type of semiconductor biosensor that measures the concentration of specific organic matter in a solution, a membrane in which an enzyme is immobilized on the surface of a semiconductor '1-field eN-immobilized membrane effect ion sensor (hereinafter referred to as 18FJGT) has been used. It is known that this is provided. This biosensor measures the concentration of a specific organic substance by using an 15FET to detect the change in hydrogen ion concentration that occurs when a specific organic substance in a solution undergoes a chemical reaction due to the catalytic action of an enzyme in an enzyme-immobilized membrane. It is something to do. As examples of enzyme-immobilized membranes with this selectivity, urease-immobilized enzyme-immobilized membranes are known for detecting urea, and glucose oxidase membranes are known for detecting glucose (Sensors and Actuators).
Actuators) Volume 7, pages 1-10 (198
5)). In addition, an 18FET (!
= A chip-based biosensor has also been developed by forming an 15FET with a deactivated enzyme-immobilized membrane on the same chip and providing a gold electrode as a reference point on the back side (No. 16, 1984 International Conference on Solid State Devices
and Materials, Late Needs Apstructs (19841international Co.
nferenceon 5olid 5tate De
vices and Materi-als, L
ate News Abstracts) 66 pages ~
67 (1984)). In manufacturing such a biosensor, it is necessary to form an enzyme-immobilized matrix on a predetermined ISFgT. Several methods are known for forming enzyme-immobilized membranes, but one method that enables the formation of enzyme-immobilized membranes at the wafer stage and enables mass production of biosensors is, for example, the method described in the cited document above. After forming an enzyme-immobilized film using cellulose triacetate as a carrier over the entire wafer surface, UV rays are irradiated through a photomask to deactivate the enzyme in the enzyme-immobilized film except on the designated 18FET. In addition, after forming an enzyme-immobilized film on a wafer coated with photoresist, a predetermined l5FE
A method has also been proposed in which the enzyme-immobilized membrane other than on the T is removed by a lift-off method (Japanese Patent Application No. 59-20916).
5).
（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った。(Problems to be Solved by the Invention) The lift-off method described above has the advantage of being able to complete the process in a short time, but the peripheral portion of the formed enzyme-immobilized film may become extremely thick.
パイＡ −１−ンサの％性は酵素固定化膜の中央部の厚
さにＩノて決定されるため、はとんどの場合このｘうｙ
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Ａう問題点があった。Since the percentage of πA-1-sa is determined by the thickness of the central part of the enzyme-immobilized membrane, in most cases this
Although the increase in thickness is not a problem in the production of biosensors,
There is a problem in that part of the thickened portion may fall off when the biosensor is used for measurement, and in rare cases may 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つのチッ
プに１つまたは２つ以上の牛専体電界効果型イオンセン
サが集積化され、そのうちの少なぐとも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法において、
Ｈ） 半導体厩界効果型イオンセンナが形成された半
導体ウェハ上に有機溶剤に可溶なフォトレジストを塗布
した後、フォトリングラフィ法によ夕酵素固定化膜が設
けられるべき所定の半纏体岨界効果型イオンセンサの表
面のフォトレジストを除く工程と、
（嗜 前記半導体つェノ＼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、
Ｉ／ウ 前記工８を経た半導体表面に酵素と架橋剤を
含む蚤白質溶液をスピン塗布して前記半導体ウェハ表面
上に酵素固定化膜を形成する工程と、に）さらに前記工
程を経た半導体ウエノ＼を前記フォトレジス）Ｋ溶解す
る有機溶剤で処理して前記フォ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ｒ、 リフトオフにより除去する工
程とを備え前記フォ） ＩＪソグラフィ法により酵素固
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７オトレジストを除く工程で、フォトレ
ジストが除かれた半導体電界効果をイオンセンサの表面
を囲むフォ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, as a means for solving the above problems, integrates one or more field-effect ion sensors dedicated to cattle in one chip, and In a method for manufacturing a semiconductor biosensor, in which an enzyme-immobilized film is provided on the surface of one semiconductor field-effect ion sensor, After applying a soluble photoresist, removing the photoresist from the surface of a predetermined semi-integrated field-effect ion sensor on which an enzyme-immobilized film is to be provided by photolithography; ＼Spinning a hydrophilic primer solution on the surface and treating the surface of the predetermined semiconductor field effect ion sensor with a hydrophilic primer; (2) forming an enzyme-immobilized film on the surface of the semiconductor wafer by spin-coating a flea white matter solution containing the above; (4) The enzyme-immobilized film should be provided by the IJ lithography method. In the step of removing the photoresist on the surface of a predetermined semiconductor field effect ion sensor, the end face of the photoresist layer surrounding the surface of the ion sensor is transferred to the surface of the semiconductor field effect ion sensor by applying the semiconductor field effect from which the photoresist has been removed. The present invention provides a method for forming an enzyme-immobilized membrane for a semiconductor biosensor, which is characterized by forming a gentle slope with respect to the membrane.
（作用）
本発明の酵素固定化膜の形成方法によれば、半導体ウェ
ハ上に有機浴剤に可溶なフォトレジストを塗布した後、
フォトリソグラフィ法によシ酵素固足化膜が設けられる
べき所定のｌ５ＦＥＴ上の表面の７オトレジストを除き
、その上から親水性プライマをスピン塗布する。フォト
レジストが除かれたｌ５ＦＩＥＴ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ＩＳＦｇＴ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蚤白質溶液を塗布するわけで
あるが、フォトリングラフィ法によってフォトレジスト
が除かれたｌ５ＦＥＴ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ェハ表面と形成
された酵素固定化膜との間に７オトレジスト層が存在す
るわけである。(Function) According to the method for forming an enzyme-immobilized film of the present invention, after coating a photoresist soluble in an organic bath agent on a semiconductor wafer,
The photoresist on the surface of a predetermined 15FET on which the enzyme-immobilized film is to be provided is removed by photolithography, and a hydrophilic primer is spin-coated thereon. The surface of 15FIET from which the photoresist has been removed is treated with a hydrophilic primer in this step to prevent the enzyme-immobilized film formed later from peeling off from the surface of ISFgT. On top of this, a flea white matter solution containing an enzyme and a cross-linking agent is applied, and an enzyme-immobilized film is formed on the surface of the 15FET from which the photoresist has been removed by the photophosphorography method, and in contact with this. In the part shown in FIG. 3, seven photoresist layers exist between the wafer surface and the formed enzyme-immobilized film.
架橋剤による架橋反応が終了するのをまりてから半導体
ウェハを有機溶剤で処理すると、フォトレジストは有機
溶剤に溶解し、その際フォ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、ｌ５ＦＪＩＧＴ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
いてフォトリングラフィ法によって所定のｌ５ＦＥＴ表
面上のフォトレジストを除いた際、１８ＦＥＴの表面を
囲むフォトレジス）４３の端面４１が第４図のように１
８Ｆ’ｇＴ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った。本発明の方法によれば、
ｌ５ＦＥＴの表面を囲むフォトレジストの端面カ第１図
１１のように１ｓＦｒｒの表面１２に対してなだらで・
、
かな傾ｇ＋をなすようにすることによ夕、す７トオ７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
のようにほぼ平坦にすることが可能となった。When the semiconductor wafer is treated with an organic solvent after the crosslinking reaction by the crosslinking agent is completed, the photoresist is dissolved in the organic solvent, and at this time, the enzyme-immobilized film formed on the photoresist is also peeled off. As a result, only the enzyme-immobilized film formed on the surface of 15FJIGT remains. In the above process, when the photoresist on the surface of a predetermined 15FET is removed by the photolithography method, the end face 41 of the photoresist 43 surrounding the surface of the 18FET becomes 1 as shown in FIG.
When perpendicular to the 8F'gT surface 42, the shape of the enzyme-immobilized membrane 51 formed as shown in FIG. 5 was significantly thicker at its periphery. According to the method of the invention,
The end face of the photoresist surrounding the surface of the 15FET is smoothed against the surface 12 of 1sFrr as shown in FIG.
The shape of the enzyme-immobilized membrane formed through the steps 7 to 7 is shown in Figure 2 21 by forming a kana slope g+.
It became possible to make it almost flat like this.
（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) Embodiments 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ナ酵素固定化膜
の形成方法の一実施例の工程説明図で、サファイア基板
上に形成された２つ一組の１８ＦＥＴの一方にのみ酵素
同定化Ｍを形成する場合−こついて示しである。第３図
（ａ）〜ｆｄ）におりて、３１はサファイア基板、３２
は高不純物＠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は窒化シリコン膜、
３６は有機溶剤可溶性の７オトレジスト層％３７は金電
極である。次に製造工程をＪ＠を追って説明する。サフ
ァイア基板表面の島状シリコン層を用いてｌ５ＦＥＴ４
形成し、サファイア基板裏面に金を蒸着したウェハの表
面に７オトレジスト・層１例工ばシラグレー社製マイク
ロボジッ＜８）
）１３００−３７をスピン塗布する（第３図（ａ））。FIG. 1 is a process explanatory diagram of an embodiment of the method for forming a semiconductor biosenna enzyme-immobilized membrane according to the present invention, in which an enzyme identification M is formed only on one of a pair of 18FETs formed on a sapphire substrate. If you do - this is a sign of trouble. In FIG. 3(a) to fd), 31 is a sapphire substrate, 32
3 is a highly impurity n-type silicon region, 33 is a p-type silicon region, 34 is a silicon oxide film, 35 is a silicon nitride film,
36 is a 7% organic solvent soluble photoresist layer, and 37 is a gold electrode. Next, the manufacturing process will be explained following J@. 15FET4 using the island-like silicon layer on the surface of the sapphire substrate
On the surface of the wafer with gold vapor deposited on the back side of the sapphire substrate, a layer of 7 photoresists (for example, Microbodies <8) 1300-37 manufactured by Silagray Co., Ltd. is spin-coated (FIG. 3(a)).
このン第１・レジストはアセトン可溶性である。次に、
フォトマスクを用いて露光を行う。このとき焼付は時の
グロキシミティーギャッン危を２０μｍＩこ設定する。This first resist is acetone soluble. next,
Exposure is performed using a photomask. At this time, the gliximity gap at the time of seizure is set to 20 μmI.
現像によ、！ｌ、ｌ酵累固疋化膜が設けられる１８ＦＥ
Ｔの表面のフォトレジスト層を除去する（第３図（ｂ）
）。以上の工程によりフォトレジストが除かれた１ｓＦ
ＥＴの表面に対しその周辺を囲むフォトレジスト層の端
面がなだかな傾斜をなすようにすることができる。その
後、親水性グライマ、例えばγ−アミノグロビルトリエ
トキシシランの１％水溶液をウェハｌこスピン塗布し、
１１０℃で５分間熱処理を行い、酵素固定化膜が設けら
れるｌ５ＦＪＩ（Ｔ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。このウェハを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、酵素と架橋剤を含む蚤白質溶液、例えば
尿素を検出する場合には３００ｍＥ／ｍｌ牛血清アルブ
ミンを含む０．１ Ｍピペラジン−Ｎ、Ｎ’−ビス（２
−エタンスルフォン酸）−水（Ｑ）
酸化ナトリウＡ （ｐＨ６，８）２体積部に５（Ｊｙｖ
ｔｉ／ｍ１のウレアーゼ（マイルスラボラトリーズ製５
３Ｕ／π４）水溶准１体績都を〃１１え、さらに２ｗｔ
優グルタルアルデヒド水浴液１体積部を加えた後よく混
合した溶液０．９７７ｚｌ ６スピン塗何する。３０分
常温で放置してグルタルアルデヒドによる架、僑反応を
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ェハ（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、フォトレジスト層はアセトン中ｌこ溶出し
、それとともにフォトレジスト層上に形成された酵素固
定化膜も剥離する。Develop it! 18FE with l,l fermentation membrane
Remove the photoresist layer on the surface of the T (Figure 3(b))
). 1sF with photoresist removed by the above steps
The end face of the photoresist layer surrounding the ET surface can be gently sloped. Thereafter, a 1% aqueous solution of a hydrophilic glimer, such as γ-aminoglobiltriethoxysilane, is spin-coated onto the wafer.
Heat treatment is performed at 110° C. for 5 minutes to bond γ-aminopropyltriethoxysilane to the surface of 15FJI (T) on which the enzyme-immobilized membrane is provided. After immersing this wafer in a 5% glutaraldehyde aqueous solution for 15 minutes, washing with water, After drying, a flea white matter solution containing enzymes and cross-linking agents, such as 0.1 M piperazine-N,N'-bis(2
- ethanesulfonic acid) - water (Q) Sodium oxide A (pH 6,8) 2 parts by volume to 5 (Jyv
ti/m1 urease (Miles Laboratories 5
3U/π4) Water-soluble quasi 1 performance capital 〃11e, further 2wt
Add 1 part by volume of the superior glutaraldehyde bath solution and apply 0.977 zl of the well-mixed solution with 6 spins. The enzyme-immobilized film 38 is formed by allowing the film to stand at room temperature for 30 minutes to complete the crosslinking reaction with glutaraldehyde. The wafer on which the enzyme-immobilized film was formed in this way (Fig. 3 (C)) was immersed in acetone and subjected to ultrasonic treatment for 2 minutes using an ultrasonic cleaner. This elution causes the enzyme immobilization film formed on the photoresist layer to also be peeled off.
その結果、あらかじめフォトレジスト膜が除去されてあ
ったｌ５ＦＢＴ上に形成された酵素固定化膜３８のみが
ウェハ上に残る（第１図り））。以上の工程により所定
のｌ５ＦＥＴの表面にだけｌと０．５μｍの均一な厚さ
の酵素固定化膜を形成することができた。As a result, only the enzyme-immobilized film 38 formed on the 15FBT from which the photoresist film has been removed remains on the wafer (first diagram). Through the above steps, an enzyme-immobilized film having a uniform thickness of 1 and 0.5 μm could be formed only on the surface of a predetermined 15FET.
（発明の効果）
辺部で肥ノｌ化することなく、均一な厚さをもって所定
のｌ５ＦＢＴ上にのみ形成される。(Effects of the Invention) The film is formed only on a predetermined 15FBT with a uniform thickness without sludge on the sides.
第１図は本発明に力）かるフォトレジスト層の断面図、
第２図は本発明の方法によっ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ォトレジスト層の
断面図、第５図は従来法によって形成された酵素固定化
膜の断面図。
図において、
１１はフォトレジストの端面、１２は１８ＦＥ’ｌ’の
表面、１３はフォトレジスト層、２１は酵素固定化膜、
２２はＩＳＰ″ＥＴの表面、３１はサファイア基板、３
２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、斜は酸化シリコン膜、３５は窒化シリコン膜、
３６はフォトレジスト層、３７は金′電極、３８は酵素
固定化膜、４１はフォトレジストの端面、４２はｌ５Ｆ
ＥＴ亭 １ 口
１３．７丁トレヴスＹ層
／２．ｌ５ＦＥＴｔ、段面
享 ２ 図
ｚｌ、謔紮圓定化譲
２２、ＩＳＦＥＴｎＲｍ
千 ３ 図
３θ＠秦固定化渠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, FIGS. 3(a) to 3(d) are process illustrations of an embodiment of the present invention, and FIG. 4 is a photo taken by the conventional method. FIG. 5 is a cross-sectional view of a resist layer, and FIG. 5 is a cross-sectional view of an enzyme immobilization film formed by a conventional method. In the figure, 11 is the end face of the photoresist, 12 is the surface of 18FE'l', 13 is the photoresist layer, 21 is the enzyme immobilization film,
22 is the surface of ISP″ET, 31 is the sapphire substrate, 3
2 is a high impurity concentration n-type silicon region, 33 is a p-type silicon region, diagonal is a silicon oxide film, 35 is a silicon nitride film,
36 is a photoresist layer, 37 is a gold electrode, 38 is an enzyme immobilization film, 41 is an end face of the photoresist, 42 is 15F
ET-tei 1 Exit 13.7 Treves Y layer/2. 15FETt, stage surface 2 Fig. zl, 謔紮圓设备22, ISFETnRm 1000 3 Fig. 3θ@Qin fixed conduit
Claims (1)
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、フォ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
を囲むフォトレジスト層の端面を前記半導体電界効果型
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。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, the enzyme-immobilized film is formed by lift-off using a photoresist. 1. A semiconductor biotechnology technology, characterized in that an end face of a photoresist layer surrounding a surface of a predetermined semiconductor field-effect ion sensor on which a chemical 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 sensor 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 true JPS62225942A (en) | 1987-10-03 |
| JPH0481740B2 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) |
Cited By (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
Cited By (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 |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0481740B2 (en) | 1992-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4894339A (en) | Immobilized enzyme membrane for a semiconductor sensor | |
| JP3105919B2 (en) | Fully microfabricated biosensor, its manufacturing method and its use | |
| US5445920A (en) | Fabrication process of biosensor | |
| JP2007139762A (en) | Biosensor, manufacturing method therefor, and method of detecting biomolecule using biosensor | |
| Hanazato et al. | Glucose sensor based on a field-effect transistor with a photolithographically patterned glucose oxidase membrane | |
| JPS61165656A (en) | Formation of immobilized enzyme membrane | |
| JPH0548418B2 (en) | ||
| JPS62225942A (en) | Method of forming semiconductor biosensor enzyme immobilized membrane | |
| JPH0349388B2 (en) | ||
| JPS62225941A (en) | Method of forming semiconductor biosensor enzyme immobilized membrane | |
| JP2946913B2 (en) | Method for producing immobilized enzyme membrane and protein immobilized membrane | |
| JPS62235556A (en) | Compound enzyme sensor | |
| JPS61283862A (en) | Manufacture of enzyme immobilized film | |
| JPS6188136A (en) | Production of semiconductor multibiosensor | |
| JPS61234349A (en) | Manufacture of semiconductor multi-biosensor | |
| JPS63241346A (en) | Manufacture of planar type biosensor | |
| JPH04173841A (en) | Method for partially forming polymer membrane having functional group | |
| JPS61274682A (en) | Production of membrane supporting immobilized enzyme | |
| JPS6250656A (en) | Biosensor and its production | |
| JPS62171684A (en) | Immobilized glucose oxidase enzyme membrane and production thereof | |
| JPS63186139A (en) | Method for selective inactivation of enzyme immobilized film | |
| JPS6186644A (en) | Manufacture of biosensor | |
| JPS63229358A (en) | Immobilized enzyme membrane and its preparation | |
| JP2687942B2 (en) | Method for forming immobilized enzyme membrane | |
| Iwama et al. | Fabrication of microchannels with patterned bio-active layers |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |