JPH10248599A - Reaction chip and its production - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a reaction chip for DNA diagnosis and therapy by drawing a stretchable base material in a linear direction or a planar direction at a specific draw ratio, allowing the base material to support the reactive substance on its micropore areas on the surface and allowing the base material to shrink to the dimension before drawing. SOLUTION: A stretchable base material as a rubber sheet is drawn in a linear or planar direction(s) bat a ratio of at least double of the length before drawing and a reactive substance as DNA fragments of the same or different kind(s), enzyme, antigen, antibody, epitope or protein are supported by extended micropore areas on the base surface. Then, the base material is allowed to shrink to the size before drawing whereby the objective reaction chip is obtained that can be used for DNA diagnosis and therapy for diseases caused by mutation of gene, for example, cancer and the like in no need of a special installation as a photolithography equipment.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a reaction chip used for DNA diagnosis and treatment, and a method for producing the same.

[0002]

2. Description of the Related Art It is known that the use of a DNA chip is effective for diagnosing diseases caused by gene mutations, for example, cancer. DNA chip is about 1cm
More than 10,000 DNA fragments (DNA
Probe). When a fluorescently labeled DNA sample to be examined is flown onto the DNA chip, the above D
DNA having sequence complementary to probe on NA chip
The fragment binds to the probe, and only that portion can be identified by fluorescence, and the sequence of the DNA fragment in the DNA sample can be elucidated. This method has already been shown to be able to detect mutations in oncogenes.

[0003]

The production of the DNA chip requires a photolithography technique used in the semiconductor industry. When the type of the DNA probe to be immobilized on the DNA chip is determined in advance, it is easy to prepare and provide such a DNA chip in a place where necessary facilities are provided. However, researchers have their own DNA
When it is desired to immobilize a probe on a DNA chip, it is necessary to prepare advanced equipment for that purpose, which is not easy. When the number of required DNA probes is small, the degree of integration of the DNA probes on the chip does not need to be so high. On the contrary, there is a case where a chip on which a desired DNA probe is immobilized more easily is desired. For example, for an organism
When the cDNA is transcribed on a chip and its generation is examined, a labeled DNA having a known specific function may be reacted as a sample.

Accordingly, an object of the present invention is to provide a DNA which can be easily prepared without requiring photolithography equipment.
An object of the present invention is to provide a reaction chip, such as a chip, having a reactive substance integrated on its surface.

[0005]

SUMMARY OF THE INVENTION The present invention relates to a reaction chip characterized in that a reactive substance is carried on at least one of a plurality of microsections on at least one surface of a stretchable substrate. . Further, the present invention provides that the stretchable substrate is stretched to enlarge the surface, the reactive substance is supported on at least one of the plurality of sections on the enlarged surface, and then the stretchable substrate is stretched before stretching. The present invention relates to a method for producing the above reaction chip, wherein the reaction chip is shrunk to a size.

[0006]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The chip of the present invention has a reactive substance carried on at least one of a plurality of microsections on at least one surface of a stretchable substrate. The stretchable substrate is not particularly limited as long as the substrate is made of a material that can be stretched at a desired magnification without being broken by stretching. Preferably, the substrate can be stretched at least twice in the stretching direction. Further, the stretchable substrate is suitably a substrate that can be stretched not only in one dimension but also in two dimensions. As such a substrate, for example, a substrate made of rubber can be mentioned. Examples of the rubber include, but are not limited to, natural rubber and silicone rubber, and are appropriately determined in consideration of various properties such as elasticity and affinity with a reactive substance on the surface of the base material. You can choose. The elasticity of the substrate used is appropriately determined according to the degree of integration of the reactive substance required for the reaction chip of the present invention. Further, the surface of the base material may be subjected to an appropriate surface treatment for the purpose of adjusting the affinity with the reactive substance and the like.

[0007] The shape of the stretchable substrate is not particularly limited. For example, it may be a flat plate such as a film or a sheet, and may be a spherical plate. Also, there is no particular limitation on the thickness and size of the substrate,
The thickness of the base material is appropriately determined in consideration of the elasticity and shape stability required for the base material, and the size of the base material is further determined in consideration of the number of micro-sections provided on the base material surface. It is determined as appropriate. In the present invention, the fine division on the substrate surface,
This is a virtual section, and each section does not exist as being physically divided, but is a section provided virtually.

[0008] The term "reactive" in the term "reactive substance" in the present invention means not only a case where a chemical reaction changes a chemical structure or the like due to an ionic bond or a covalent bond, but also a hydrogen bond, a coordinate bond, a Van der Waals force. , Chemical adsorption, physical adsorption, etc., that can create a state of being combined with other substances. Examples of such a reactive substance include, but are not limited to, DNA fragments, enzymes, antigens, antibodies, epitopes, proteins, and the like.

[0009] In the reaction chip of the present invention,
That is, there is no particular limitation on the degree of accumulation of reactive substance categories. The required and convenient degree of integration differs according to the use of the reaction chip, and therefore the degree of integration can be changed as appropriate according to the use. Exemplarily, the number of micro-sections per 1 cm ^{2 of} the reaction chip surface can be 100 or more. By adjusting the material of the base material and the reactive substance, about 10000 micro-sections per 1 cm ² of the surface can be obtained. It can be provided.

The reaction chip of the present invention is characterized in that a stretchable base material is stretched to enlarge a surface, a reactive substance is supported on at least one of a plurality of sections on the enlarged surface, and then the stretchable base is stretched. It can be produced by shrinking the material to the size before stretching. The stretching of the substrate can be performed one-dimensionally or two-dimensionally. The stretching method is not particularly limited. For example, in the case of a rectangular (eg, square, rectangular) sheet-like base material, when one-dimensionally or two-dimensionally stretched, each end of the sheet is stretched along opposite sides. Fix the part to an appropriate jig,
The jig can be stretched by moving the jig in the direction in which the sheet is stretched. When the substrate is spherical (hollow balloon type), the surface can be stretched to a desired size by injecting a gas or liquid such as air into the hollow of the spherical substrate and inflating the same. . There is no particular limitation on the degree of stretching,
It can be appropriately determined in consideration of the degree of accumulation of the reactive substance and the workability of supporting the reactive substance. For example, the length in each stretching direction can be stretched twice or more before stretching.

A reactive substance is carried on each of the enlarged micro sections on the surface of the substrate. The reactive substances can be the same or different substances, depending on the application of the reaction chip. Further, from the viewpoint of work efficiency, it is preferable to carry a plurality of reactive substances at once, and more preferably to carry all the reactive substances at once. The reactive substance to be supported can be supported on the surface of the substrate by the reactive substance alone, but if necessary, the reactive substance is dissolved or dispersed in an appropriate solvent, and the solution or dispersion is applied to the fine particles on the surface of the substrate. It is also possible to drip or adhere to the section and then remove the solvent if necessary. Solvents that dissolve or disperse the reactive substance
It can be appropriately selected in consideration of compatibility with the reactive substance, affinity with the substrate surface, and the like. The chip of the present invention can be obtained by shrinking the substrate supporting the reactive substance to the size before stretching.

[0012]

According to the present invention, it is possible to easily provide a reaction chip in which a reactive substance such as a DNA fragment is integrated on the surface thereof without requiring special equipment such as photolithography equipment. Further, by selecting a base material of a material having high elasticity and / or devising a method of supporting a reactive substance, a chip having a higher integration degree than a DNA chip using existing silicon is provided. It is also possible.

[0013]

The present invention will be further described below with reference to examples. Example 1 A rubber sheet having a side of 3 cm was two-dimensionally (XY) as shown in FIG.
Each film was stretched 5 times in both directions. For this stretching, a stretching device provided with a sheet holding jig shown in FIG. 2 was used. On the surface of a two-dimensionally stretched sheet (about 15 cm × 15 cm), the reactive substance supporting device shown in FIG. 3 [a plurality of (26 × 26) ultrafine capillaries (diameter of each capillary 0.4 mm, distance between centers of capillaries) 1 mm)] is filled with a solution containing a DNA fragment), and the solution containing the DNA fragment is supported on the sheet surface so that each solution does not come into contact with another solution. . After evaporating the solvent in the solution, the sheet was shrunk to the size before stretching to obtain a reaction chip of the present invention. About 2.5cm 676 in the corner
A reaction chip in which dots of each DNA fragment were independently carried was obtained.

Example 2 In a normal state, air or liquid is injected under pressure into a rubber balloon having a diameter of 5 cm to expand the balloon to a diameter of about 25 cm, and the expanded balloon is placed on two plastic plates (the center of one plastic plate). (There is an opening of 4 cm x 3 cm in the part).
Reactive substance supporting pins [plural (40 × 30
Book), each pin diameter 0.3mm, pin center distance 1mm)]
The solution containing the enzyme was placed on the pin tip of the device having the above. The tip of the pin was brought into contact with the portion (substantially flat) exposed to the opening of the balloon, and the solution containing the enzyme was carried on the exposed surface so that each solution did not come into contact with other solutions. . After evaporating the solvent in the solution, the balloon was contracted to the size before expansion (stretching), and the reaction chip of the present invention was obtained as it was or by cutting off the portion supporting the enzyme. A reaction chip having 1200 enzyme dots independently supported on about 4 cm × 3 cm was obtained.

[Brief description of the drawings]

FIG. 1 is an explanatory view of a method for producing a reaction chip of the present invention.

FIG. 2 is an explanatory view of a stretching device provided with a sheet holding jig.

FIG. 3 is an explanatory view of an apparatus for supporting a reactive substance.

FIG. 4 is an enlarged view of a reactive substance-supporting pin (one).

Claims (8)

[Claims] 1. A reaction chip wherein a reactive substance is carried on at least one of a plurality of micro sections on at least one surface of an elastic substrate. 2. The reaction chip according to claim 1, wherein the stretchable base material is a base material that can be stretched at least twice in the stretching direction without breaking by stretching. 3. The reaction chip according to claim 1, wherein the elastic substrate is a substrate that can be stretched two-dimensionally. 4. The reaction chip according to claim 1, wherein the elastic substrate is a rubber substrate. 5. The reactive substance is a DNA fragment, an enzyme, an antigen,
5. An antibody, epitope or protein.
The reaction chip according to any one of the above. 6. Minute sections per cm ² of at least 100
The reaction chip according to any one of claims 1 to 5, wherein the number is one. 7. A stretchable base material is stretched to enlarge the surface,
The reaction chip according to claim 1, wherein the reactive substance is supported on at least one of the plurality of sections on the enlarged surface, and then the elastic substrate is contracted to a size before stretching. Method. 8. The stretchable base material is stretched one-dimensionally or two-dimensionally, and the length in each stretching direction is at least twice as long as that before stretching, and the same or different kind of each of the enlarged minute sections on the surface of the base material. The method according to claim 7, wherein the reactive substance is supported at one time, and then the substrate is shrunk to a size before stretching.