JPS61234349A - Manufacture of semiconductor multi-biosensor - Google Patents

Abstract

PURPOSE:To form a biosensor having several kinds of enzyme immobilized films, by a method wherein an enzyme immobilized film and a photoresist layer are formed on a substrate provided with an ion sensor and a process of removing all but a portion required therefor. CONSTITUTION:An ion induction section 2 of an FE type ion sensor is provided on the top surface of a substrate 1 and an enzyme immobilized film 3 and a photoresist layer 4a are formed over the entire surface thereof. Then, the photoresist layer 4a is removed except for a specified surface of the ion induction section 2a by exposure and development and the enzyme immobilized film 3 is decomposed. Thereafter, a second enzyme immobilized film 5 and a photoresist layer 4b are formed on the top surface of the ion induction section and the same removing process is repeated to form a biosensor having different enzyme immobilized films 3 and 5 on the ion induction section 2. Thus, a massproducible fine multi-biosensor can be obtained by applying an IC production technology.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a semiconductor multi-biosensor that is formed by integrating semiconductor ion sensors each having an enzyme-immobilized membrane provided on its surface.

[Conventional technology]

Ion sensitive field effect transistor (ZonS@na
lt1vs F1@ld Eff@ct Transi
The /4 iosensor, which has an enzyme-immobilized membrane formed on the ion-sensitive membrane surface of the stor l5FET, is a silicon ICtll.
Since it is manufactured using the same manufacturing technology, it is possible to integrate multiple sensors.

The present inventors have developed a method for forming different kinds of enzyme-immobilized membranes on the ion-sensitive membrane surfaces of multiple 15FETs by coating the parts other than the ion-sensitive parts with a hydrophobic resin in advance, and then dropping an enzyme-containing solution dropwise. He proposed a method for forming an enzyme membrane (Japanese Patent Application No. 208626/1983).

[Problems that the invention is supposed to solve]

According to this method, various types of enzyme membranes can be formed in a short time, but fine mechanical operations are required. Furthermore, the present inventors
As a means to overcome this drawback, we proposed a method of patterning a plurality of enzyme membranes by repeating photolithography (Japanese Patent Application No. 59-209166). Although this method made it possible to perform arm turning using a normal IC process, there remained a problem in its applicability when the thickness of the film exceeds a certain level.

It is an object of the present invention to provide a method for manufacturing a semiconductor multi-biosensor suitable for mass production, which eliminates such conventional problems and allows different enzyme membranes to be easily formed in the ion-sensing parts of different 15FETs.

[Means for solving problems]

The present invention relates to a method for manufacturing a semiconductor multi-biosensor that integrates semiconductor ion sensors having a plurality of different enzyme-immobilized films on the surface. a step of forming a photoresist layer on the enzyme-immobilized film in the area that is exposed to the ion-sensitive area of a predetermined ion sensor; This method of manufacturing a semiconductor multi-ion sensor is characterized by forming a plurality of different enzyme-immobilized films by repeating the step of decomposing the enzyme film, and then removing the photoresist layer.

〔Example〕

An embodiment of the present invention will be described in detail below with reference to the drawings.

FIGS. 1(&) to 1(1) are diagrams for explaining one embodiment of the method for manufacturing a semiconductor biosensor according to the present invention, and are cross-sectional views of the ion-sensitive portion of senna in each step. This figure shows the case where different enzyme films are formed on two 15FETs on a sapphire substrate.

'In Figures 1 (a) to (1), 1 is a sapphire substrate, 2 is an ion sensitive part of 18FET, 3 is a first enzyme immobilization membrane, 4 is a photoresist layer, and 5 is a second enzyme immobilization layer. It is a membrane. Next, the KJIIi manufacturing process will be explained step by step.

A first
A protein solution containing an enzyme and a crosslinking agent (first enzyme-immobilized membrane 3)
) is applied by spin coating (Fig. 1(a)).

For example, when detecting urea, add i! ia x00trvA
d-urease (Peringer Mannheim H) approx. so
A solution obtained by adding 250 μt of η~ solution and adding 500 μt of a 0.75% glutaraldehyde aqueous solution and mixing is used. Next, an acetone-soluble photoresist, such as AZ 1450 J manufactured by Silapray, is spin-coated on the surface of the enzyme immobilization film 30 (photoresist layer 4&) (see Fig. 1(b).
)).

The photoresist film 4t is removed from the portion other than the surface of the ion-sensitive portion 2a of 15FKT provided on the first enzyme immobilization film 3 by exposure using a photomask.
Figure (C)). This wafer is immersed in Ichigintsuku's trypsin aqueous solution for 1 minute to decompose the enzyme-immobilized membrane 3 (see Figure 1).
d)). Next, a protein solution containing a second enzyme and a crosslinking agent is spin coated (second oxygen fixing film 5). For example, when detecting glucose, add 501n9Al glucose oxidase (
Manufactured by Peringer Mannheim, approx. 250″M7fq)
) solution, 500 μt of 0.75% glutaraldehyde aqueous solution was added thereto, and a mixed solution was used. On top of this, apply an acetone-soluble photoresist (
Photoresist layer 4b) (FIG. 1C@)). The photoresist layer 4b in the portion other than the surface of the ion-sensitive part 2b of the 15FET provided on the second enzyme immobilization film 5 is removed by exposure and development using a photomask (see Fig. 1(f)).
)). Soak the quackfish in one beak of trypsin aqueous solution,
The second oxygen fixation membrane 5 is decomposed. After this, the photoresist layer 4 was immersed in a trypsin inhibitor aqueous solution similar to 1 Gin to stop the protein decomposition reaction, and then immersed in acetone.
(Fig. 1 (h)). Cut the wafer and take out the individual sensors (Fig. 1 (1)).

The thickness of the enzyme-immobilized film obtained by this method can be easily controlled by controlling the protein concentration and rotation speed during spin coating. In the example, 3,000”pms 10
It was possible to obtain a uniform enzyme-immobilized film with a thickness of 50,000 mm or less by spin coating for seconds.

Further, in this case, although the adhesion of the enzyme membrane to the ion-sensitive membrane was good, it is also possible to perform primer treatment in advance to further improve the adhesion.

〔Effect of the invention〕

According to the present invention, mass production is possible using IC manufacturing technology, and a microscopic multi-biosensor can be obtained.

It is clear that by repeating the steps of the present invention, 15FETs each having three or more types of enzyme-immobilized membranes formed on their surfaces can be provided on the same chip.

It is also clear that enzyme turning can be performed by this method not only on 15FETK but also on biosensors using minute amberometric electrodes.

[Brief explanation of the drawing]

FIGS. 1(a) to 1(1) are diagrams for explaining one embodiment of the method for manufacturing a semiconductor Δ-iosensor according to the present invention, and are cross-sectional views of the ion-sensing portion of the ion sensor in each step. 1... Sapphire substrate, 2 (2m, 2b)...
- Ion sensitive part of ion sensor, 3... First enzyme immobilization membrane, 4a r 4b... Photoresist layer, 5.
...Second enzyme immobilization membrane. Figure 1Figure 1

Claims (1)

[Claims] (1) In a method for manufacturing a semiconductor multi-biosensor that integrates semiconductor ion sensors with multiple different enzyme-immobilized membranes on the surface, the enzyme-immobilized membrane is coated on a semiconductor wafer by applying a protein solution containing an enzyme and a crosslinking agent. a step of forming;
Repeating the process of forming a photoresist layer on the enzyme-immobilized membrane in the area corresponding to the ion-sensitive area of a predetermined ion sensor, and the process of decomposing the enzyme membrane in the area excluding the ion-sensitive area using a protease. 1. A method for manufacturing a semiconductor multi-ion sensor, comprising forming a plurality of different enzyme-immobilized films and then removing a photoresist layer.