JP2005172711A - Manufacturing method of microarray chip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a manufacturing method of a microarray chip capable of correcting a defective part or removing the same when the defective part is formed and having a specific biopolymer arranged and fixed thereto in a fine patternwise state without causing the non-specific adsorption of an object to be inspected or the like. <P>SOLUTION: The manufacturing method of the microarray chip has a specific biopolymer fixing process for fixing a specific biopolymer on a base material and an organic group removing process for removing the organic group present in the space part between specific biopolymer fixing parts. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a method for producing a microarray chip in which a large number of specific biopolymers that specifically bind to specific DNA or protein are arranged.

  In recent years, DNA chips, DNA arrays, protein chips, capillary arrays, μTAS, MEMS, etc. that can perform high-throughput biological detection of many types or functions (biological interact), such as nucleic acid and amino acid sequences on the substrate A microarray chip having a specific biopolymer immobilized thereon has been proposed and put into practical use. In the production of these microarray chips, wafer processing technology, microcontact printing technology, ink jet technology, etc., which are also used in semiconductor production, have been incorporated, and various specific biopolymers can be accurately arranged in a minute area. By fixing, it is possible to detect from a very small amount of sample in a short time.

  For example, a DNA chip in which an oligonucleotide or cDNA (DNA complementary to mRNA (a nucleic acid encoding protein sequence information)) that is part of deoxyribonucleic acid (DNA) is arrayed and immobilized as a specific biopolymer on a substrate And a microarray chip called a DNA array. Examples of nucleic acids arranged on such a DNA chip or DNA array include short-chain (20 to 30 bases) and long-chain (50 bases or more) oligonucleotides and cDNA. As a method for arranging and immobilizing these nucleic acids, a photopolymer synthesis method is used when the nucleic acid is a short oligonucleotide, and when cDNA is used as the nucleic acid, microcontact printing or inkjet technology is used. A method using the used array device is generally used. When long-chain oligonucleotides are used as nucleic acids, both are used for a while.

  Here, in the method by mechanical processing using microcontact printing or ink jet technology used when arranging and immobilizing cDNA, etc., preparing multiple types of specific biopolymers prepared in advance, The specific biopolymer is spotted at a high density on the substrate and solidified, whereby the specific biopolymer can be immobilized and a microarray chip can be obtained. As such a method, for example, P. of Stanford University. Examples include the method developed by Brown et al.

  In graphic prints and the like using the above-described array device, it has been established that printing is performed at 600 dpi or more. However, as described above, when spotting a specific biopolymer, adjacent dots (specific biomaterials) are used. ) It is required that the materials are not mixed with each other, and the bleeding of the substrate cannot be suppressed, so that it is only about 120 dpi. Therefore, when a specific biopolymer is spotted in a fine pattern in order to obtain high throughput, the spotted specific biopolymer diffuses and the adjacent specific biopolymer comes into contact. In some cases, the specific biopolymer adheres to the space between adjacent specific biopolymers.

  However, when different specific biological substances are mixed in this way, or when a specific biopolymer adheres to the space part, it is difficult to remove or modify the part. The test object was non-specifically adsorbed, which caused the contrast of each specific biological material to decrease and the accuracy to decrease.

  Therefore, when a defective portion occurs, the specific biopolymer can be corrected and removed, and the specific biopolymer can be formed in a high-definition pattern without causing non-specific adsorption of the test object. There is a need to provide a method for manufacturing an arrayed and fixed microarray chip.

The present invention includes a specific biopolymer immobilization step for immobilizing a specific biopolymer on a substrate,
And an organic group removing step of removing an organic group present in a space between the specific biopolymer immobilization parts. A method for producing a microarray chip is provided.

  According to the present invention, the organic group in the space portion between the specific biopolymer immobilization portions to which the specific biopolymer is finally immobilized is removed by the organic group removal step. Can be a microarray chip to which no organic group such as a specific biopolymer is attached. As a result, the object to be inspected cannot be non-specifically adsorbed on the space portion of the microarray chip manufactured according to the present invention, and the microarray chip has high contrast and high accuracy. It can be done.

The present invention also includes a specific biopolymer immobilization step for immobilizing a specific biopolymer on a substrate,
There is provided a method for producing a microarray chip, comprising: a defective portion removing step of removing a defective portion of the specific biopolymer when a defect occurs in the specific biopolymer immobilization step.

  According to the present invention, in the specific biopolymer immobilization step, when the different specific biomaterial is formed, or when different specific biopolymers are mixed, the defective portion These defective portions can be removed by the removing step. Therefore, it is possible to obtain a high quality microarray chip in which the object to be inspected is not non-specifically adsorbed on the defective portion.

  At this time, a second specific biopolymer immobilization step of immobilizing the specific biopolymer may be included in the defective portion removal portion from which the defective portion has been removed by the defective portion removal step. This is because the target specific biopolymer can be immobilized again on the defective portion removing portion from which the defective portion has been removed, and a high-quality microarray chip can be obtained.

  In the above invention, the base material has a characteristic change layer whose characteristics change due to the action of the photocatalyst accompanying energy irradiation, and the photocatalyst-containing layer side substrate having the photocatalyst-containing layer and the substrate containing the photocatalyst is contained. It is preferable to have a characteristic changing step of changing the characteristic of the specific biopolymer immobilization part on the characteristic changing layer by irradiating energy after arranging the layer with a gap from the characteristic changing layer. . This makes it possible to adhere the specific biopolymer-containing coating liquid by utilizing the difference in characteristics such as wettability of the characteristic change layer, and easily attach the specific biopolymer to a high-definition pattern. This is because it can be fixed in a shape.

  In this case, the characteristic change layer can be a wettability change layer in which the wettability is changed so that the contact angle with water is lowered by the action of the photocatalyst accompanying energy irradiation. In this case, the specific biopolymer immobilization part can be a lyophilic area, and the area other than the specific biopolymer immobilization part can be a liquid-repellent area. It can be easily attached only on the polymer fixing part. Thereby, it can prevent that the adjacent specific biopolymer is mixed or contacted.

  At this time, the characteristic change layer can be a decomposition / removal layer that is decomposed and removed by the action of the photocatalyst accompanying energy irradiation. In this case, it is possible to easily apply the specific biopolymer-containing coating liquid by using unevenness due to the presence or absence of the decomposition removal layer, and to prevent mixing of adjacent specific biopolymers. is there.

  In the above invention, the organic group removal step or the defective portion removal step may include the step of removing the photocatalyst-containing layer of the photocatalyst-containing layer side substrate having the photocatalyst-containing layer and the substrate. It is preferable to carry out by irradiating energy after arranging it. According to this method, since the organic group removal step and the defective portion removal step can be performed by the action of a photocatalyst accompanying energy irradiation, for example, in the case of using a photolithography method, a specific biopolymer or the like is used. There is no need to use chemicals that have adverse effects. In addition, by irradiating the space portion and the defective portion with energy using the photocatalyst-containing layer side substrate, the organic group or the like can be easily decomposed and removed, so that the above process can be performed efficiently. It is because it has.

  The present invention also provides a microarray chip comprising a substrate and a specific biopolymer fixed in a pattern on the substrate.

  According to the present invention, since the specific biopolymer is formed in a pattern, the test object does not adsorb nonspecifically between adjacent specific biopolymers. A microarray chip capable of performing accurate detection can be obtained.

  In the said invention, it is preferable that the said specific biopolymer is formed on the characteristic change layer in which a characteristic changes by the effect | action of the photocatalyst accompanying energy irradiation formed on the base material. In this case, the specific biopolymer can be easily formed by utilizing the difference in characteristics of the characteristic change layer.

  Moreover, in the said invention, the layer which consists of polysiloxane which has a hydroxyl group at the terminal may be formed. This is because even when such a layer is formed, a high-quality microarray chip can be obtained without non-specifically adsorbing the inspection object in the space portion.

  According to the present invention, the organic group in the space portion between each specific biopolymer immobilization portion is removed by the organic group removal step, so that an organic group such as a specific biopolymer adheres to the space portion. It can be set as the microarray chip which is not. As a result, the object to be inspected cannot be non-specifically adsorbed on the space portion of the microarray chip manufactured according to the present invention, and the microarray chip has high contrast and high accuracy. There is an effect that it is possible.

The present invention can correct even when a defective part is produced during production, and the specific biopolymer is arranged and fixed in a fine pattern without non-specific adsorption of the test object. The present invention also relates to a method for manufacturing a microarray chip and a microchip.
Each will be described below.

A. First, a method for manufacturing a microarray chip of the present invention will be described. There are two embodiments of the microarray chip of the present invention. Hereinafter, each embodiment will be described separately.

1. First Embodiment First, a first embodiment in the method for manufacturing a microarray chip of the present invention will be described. The method for producing a microarray chip of this embodiment includes a specific biopolymer immobilization step of immobilizing a specific biopolymer on a substrate,
And an organic group removing step of removing an organic group present in the space between the specific biopolymer immobilization parts.

  For example, as shown in FIG. 1, the microarray chip manufacturing method of the present embodiment allows a specific biopolymer-containing coating solution to adhere to a specific biopolymer fixing portion a and solidify on a substrate 1. Specific biopolymer immobilization step (FIG. 1 (a)) for making specific biopolymer 2 and specific biopolymer 2 etc. existing in the space b between the specific biopolymer immobilization parts a And an organic group removing step (FIG. 1B) for removing the organic group. Here, the specific biopolymer immobilization part is a region where the specific biopolymer is finally immobilized in the microarray chip. The space portion refers to a region between adjacent specific biopolymer immobilization portions adjacent to each other, and includes a region between the specific biopolymer and the specific biopolymer of the manufactured microarray chip. To do.

  In general, when the specific biopolymer-containing coating solution is adhered onto the specific biopolymer immobilization part by mechanical means such as an inkjet method, the specific biopolymer-containing coating solution is wetted. It spreads and the specific biopolymer may adhere to the space part. In such a case, when the inspection object is detected using the microarray chip, the non-inspection object is non-specifically adsorbed on the space portion, and there is a problem that the detection cannot be performed with high accuracy. .

  Therefore, in this embodiment, an organic group removal step for removing the organic group on the space portion is performed, and nonspecific adsorption does not occur between the organic group present in the space portion and the test object. It is a microarray chip.

  Hereafter, each process in the manufacturing method of the microarray chip | tip of this embodiment as mentioned above is demonstrated.

(Specific biopolymer immobilization process)
First, the specific biopolymer immobilization step in this embodiment will be described. In the specific biopolymer immobilization step in this embodiment, the specific biopolymer-containing coating solution containing the specific biopolymer is attached to the specific biopolymer immobilization part on the base material and solidified. This is a step of immobilizing the specific biopolymer.

  Here, in this step, the method for immobilizing the specific biopolymer is not particularly limited, and a method for immobilizing the specific biopolymer can be used in a general microarray chip. . Specifically, a specific biopolymer-containing coating solution containing a specific biopolymer can be immobilized by adhering it to a target region and solidifying it. The specific biopolymer-containing coating solution used in such a method is a coating solution containing the above specific biopolymer, and usually the specific biopolymer was dissolved in water or an aqueous solvent. Is. Here, the specific biopolymer refers to a biopolymer that can specifically bind to a predetermined biopolymer, and the specific biopolymer used in this step has such properties. It is not particularly limited as long as it has a biopolymer.

  Examples of such a combination of a specific biopolymer and a biopolymer include, for example, a combination of a nucleic acid (eg, oligonucleotide or polynucleotide) and a complementary nucleic acid, an antigen and an antibody (or antibody fragment) Combinations, combinations of receptors and their ligands (eg, hormones, cytokines, neurotransmitters, or lectins), combinations of enzymes and their ligands (eg, enzyme substrate analogs, coenzymes, modulators, or inhibitors) And a combination of an enzyme analog and a substrate of the enzyme that is the base of the enzyme analog, or a combination of a lectin and a sugar. The “enzyme analog” means a substance having a high specific affinity with a substrate for the original enzyme but showing no catalytic activity. In addition, each of the compounds in each of the above combinations is a “specific biopolymer” and the other is a substance to be detected. For example, in the “combination of antigen and antibody”, when the antigen is “a substance to be detected”, the antibody can be “specific biopolymer”, and conversely, the antibody is “detected” In the case of “substance”, the antigen becomes “specific biopolymer”.

  For example, when the microarray chip produced according to this embodiment is applied to a nucleic acid hybridization assay, the specific biopolymer is complementary to a nucleic acid (for example, an oligonucleotide or a polynucleotide) that is a substance to be detected. Oligonucleotides or polynucleotides that can be bound to each other can be used. As used herein, “oligonucleotide” or “polynucleotide” includes 2′-deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and peptide nucleic acid (PNA). PNA is an artificial nucleic acid obtained by converting a DNA phosphodiester bond into a peptide bond. The chain length of the oligonucleotide or polynucleotide bound to the insoluble particle can be appropriately selected depending on the purpose of analysis, and is based on, for example, the chain length of the complementary sequence of DNA, RNA, or PNA to be captured. Can be determined. In addition, by arranging oligomers of confirmed base sequences in different combinations on the array and determining the complementary binding position with a part of the specific biopolymer that is sufficiently longer than that, the sequence can be determined by a computer. It is also possible to perform a wide range of sequencing by mining common points.

Moreover, when the microarray chip of this embodiment is applied to an immunological assay, an antigen (including a hapten) or an antibody that specifically binds to a substance to be detected may be used as the specific biopolymer. it can. In this case, the substance to be detected is not particularly limited as long as it is a component generally contained in a test sample and can be detected immunologically. For example, various proteins, polysaccharides, lipids, fungal bodies, various environmental substances, and the like can be given. More specifically, an immunoglobulin (eg, IgG, IgM, or IgA), an infection-related marker (eg, HBs antigen, HBs antibody, HIV-1 antibody, HIV-2 antibody, HTLV-1 antibody, or treponema antibody) Tumor associated antigens (eg, AFP, CRP, or CEA), coagulation fibrinolytic markers (eg, plasminogen, antithrombin-III, D-dimer, TAT, or PPI), antiepileptic drugs (eg, hormones), Examples include various drugs (for example, digoxin), bacterial cells (for example, O-157 or Salmonella) or their endotoxins or exotoxins, microorganisms, enzymes, residual agricultural chemicals, environmental hormones, and the like. As the antibody used as the specific biopolymer, either a polyclonal antibody or a monoclonal antibody obtained by a well-known method can be used. Further, the antibody may be a protein [for example, an enzyme (for example, pepsin or papain)] [for example, an antibody fragment (for example, Fab, Fab ′, F (ab ′) ₂ , or Fv)]. it can.

  In this step, the specific biopolymer-containing coating solution containing such a specific biopolymer is adhered to the specific biopolymer fixing part on the base material and solidified, thereby allowing the specificity. A biopolymer can be immobilized. In this step, the specific biopolymer is immobilized in this step in a wider area than the specific biopolymer immobilization part that finally immobilizes the specific biopolymer, and organic group removal described later is performed. In the step, the specific biopolymer in a region other than the specific biopolymer immobilization part may be removed.

  Here, as described above, the specific biopolymer immobilization part is an area where the specific biopolymer immobilization is performed, and a specific biopolymer different from each specific biopolymer immobilization part is usually immobilized. It will be The shape of the specific biopolymer immobilization part is appropriately selected depending on the use of the microarray chip, and may be, for example, a circle or the like, but is generally a rectangular pattern.

  Also, depending on the use of the microarray chip, etc., when the specific biopolymer is immobilized using the specific biopolymer-containing coating solution as in the present embodiment, the above-mentioned specific is usually on one microarray chip. There are about 100 to 200,000 sex biopolymer immobilization parts, especially about 400 to 50,000. At this time, the space portion, which is a region between the adjacent specific biopolymer immobilization portions, is usually 1 μm to 2,000 μm, particularly 10 μm to 400 μm, particularly 20 μm to 200 μm.

  Here, the method for applying the specific biopolymer-containing coating solution is not particularly limited as long as it is a method capable of applying the specific biopolymer-containing coating solution. A method used for manufacturing a microarray chip can be used. Specific examples include nozzle discharge means including a microcontact printing method, an ink jet method, and a method using a dispenser, electrospray (electrostatic spraying method), and the like.

  In addition, the shape of the base material used in this step is not particularly limited as long as the specific biopolymer-containing coating liquid can be attached, and a base material on a flat plate is used. However, for example, a concave portion may be formed in the specific biopolymer adhesion portion. This is because the specific biopolymer-containing coating liquid does not wet around and spreads the specific biopolymer with high definition.

  As the substrate used in this step, a substrate usually used for a microarray chip can be used. For example, silicon wafer, metal, quartz, glass, diamond, diamond-like carbon (DLC), alumina, polymer material Etc. can be used, and these are appropriately selected and used according to the application of the microarray chip to be manufactured.

  Further, in this embodiment, the substrate may have an immobilization layer or the like on the surface in order to more firmly fix the specific biopolymer, and the above specific biopolymer-containing coating solution In order to attach the high-definition on the specific biopolymer immobilization part, it may have a characteristic change layer whose characteristic changes due to the action of the photocatalyst accompanying energy irradiation.

Here, in this embodiment, in particular, the substrate has a property changing layer whose properties change due to the action of the photocatalyst accompanying energy irradiation, and the method for manufacturing the microarray chip of this embodiment will be described later. It is preferable to have a characteristic changing step of changing the characteristic of the characteristic changing layer in the pattern of the specific biopolymer immobilization part. Thereby, the specific biopolymer-containing coating liquid can be applied along the pattern in which the characteristics of the characteristic change layer have changed, and the specific biopolymer wets and spreads, and the adjacent specific biopolymer This is because it can be prevented from coming into contact with or mixing. In addition, when such a characteristic change layer is used, the organic group of the characteristic change layer can be easily removed in the organic group removal step described later, and the surface can be made lyophilic or decomposed and removed. Therefore, it is possible to obtain a high-quality microarray chip in which the object to be inspected does not adsorb unspecifically in the space portion.
Since the immobilization layer, the characteristic change layer, and the characteristic change process will be described in detail later, a detailed description thereof is omitted here.

(Organic group removal process)
Next, the organic group removal step in this embodiment will be described. The organic group removal step in this embodiment is a step of removing organic groups present in the space portion between the specific biopolymer immobilization portions. This organic group removal step is usually performed after the specific biopolymer immobilization step.

  Here, as the organic group to be removed by this step, the specific biopolymer wetted and spread on the space part, or when the substrate has the immobilization layer or the property change layer, these Examples include the organic group of the layer. When a specific biopolymer is attached to the space part, or when an immobilization layer, a characteristic change layer, or the like is formed, when the manufactured microarray chip is used, it is covered on the space part. The test object may adsorb non-specifically, leading to a decrease in accuracy. Therefore, by removing these organic groups in this step, the test object does not adsorb unspecifically in the space portion of the microarray chip, and a highly accurate microarray chip can be obtained.

  Here, the method for removing the organic group in this step is not particularly limited as long as it is a method capable of removing the organic group present in the space portion. For example, it may be performed by a commonly performed photolithography method or the like, or may be a method of decomposing an organic group by irradiating, for example, short wavelength ultraviolet light or the like in a pattern. .

  In the present embodiment, for example, as shown in FIG. 2, a photocatalyst containing layer side substrate 13 having a photocatalyst containing layer 11 and a base 12 is prepared, and the above-mentioned specific living body height formed on the base material 1 is prepared. After disposing the molecule 2 and the photocatalyst-containing layer 11 with a gap between them, the space 15 between adjacent specific biopolymer adhering portions a is irradiated with energy 15 using, for example, a photomask 14 or the like. Preferably, it is done.

According to this method, the organic group can be removed by the action of a photocatalyst accompanying energy irradiation, so that it adversely affects a specific biopolymer such as an alkali developer used in a photolithography method. There is no need to use special chemicals. In addition, by irradiating the space portion with energy using the above-mentioned photocatalyst-containing layer side substrate, it is possible to easily decompose and remove organic groups, etc., so that organic groups can be efficiently removed without using high energy light. This is because it also has the advantage that it can be performed.
The photocatalyst containing layer side substrate, energy, etc. used in the method for removing organic groups using such a photocatalyst containing layer side substrate will be described below.

a. Photocatalyst containing layer side substrate First, the photocatalyst containing layer side substrate used in this step will be described. The photocatalyst containing layer side substrate used in this step has a photocatalyst containing layer containing a photocatalyst and a substrate. Thus, the photocatalyst-containing layer side substrate used in this step has at least a photocatalyst-containing layer and a substrate, and usually a thin-film photocatalyst-containing layer formed by a predetermined method is formed on the substrate. It will be. Moreover, the thing in which the photocatalyst containing layer side light shielding part formed in pattern shape was formed can also be used for this photocatalyst containing layer side board | substrate. Hereinafter, each structure of the photocatalyst containing layer side substrate will be described.

(1) Photocatalyst-containing layer The photocatalyst-containing layer used in this step is not particularly limited as long as the photocatalyst in the photocatalyst-containing layer can decompose and remove the target organic group. The photocatalyst and the binder may be used, or the photocatalyst may be formed into a single film. Further, the wettability of the surface may be particularly lyophilic or lyophobic.

  The photocatalyst-containing layer used in this step may be formed on the entire surface of the substrate 12 as shown in FIG. 2, for example, but for example, as shown in FIG. 11 may be formed in a pattern.

  By forming the photocatalyst-containing layer in a pattern in this way, it is not necessary to irradiate energy in a pattern using a photomask or the like at the time of energy irradiation, which will be described later. This is because part of the organic group can be removed.

  In addition, since only the organic groups that actually face the photocatalyst-containing layer are decomposed and removed, the energy irradiation direction is such that the energy is directed to the part where the photocatalyst-containing layer and the specific biopolymer etc. face each other. As long as it irradiates, you may irradiate from what direction, and also has the advantage that the irradiated energy is not limited to parallel things, such as parallel light.

  In such a photocatalyst-containing layer, the mechanism of action of a photocatalyst represented by titanium dioxide as described later is not necessarily clear, but a carrier generated by light irradiation reacts directly with a nearby compound, or It is considered that the active oxygen species generated in the presence of oxygen and water change the chemical structure of organic matter. In the present invention, it is considered that this carrier acts on the organic group in the space portion arranged in the vicinity of the photocatalyst containing layer.

Examples of the photocatalyst used in the present invention include titanium dioxide (TiO ₂ ), zinc oxide (ZnO), tin oxide (SnO ₂ ), strontium titanate (SrTiO ₃ ), tungsten oxide (WO ₃ ), which are known as photo semiconductors. Bismuth oxide (Bi ₂ O ₃ ) and iron oxide (Fe ₂ O ₃ ) can be mentioned, and one or a mixture of two or more selected from these can be used.

  In this embodiment, titanium dioxide is particularly preferably used because it has a high band gap energy, is chemically stable, has no toxicity, and is easily available. Titanium dioxide includes an anatase type and a rutile type, and both can be used in the present invention, but anatase type titanium dioxide is preferred. Anatase type titanium dioxide has an excitation wavelength of 380 nm or less.

  Examples of such anatase type titanium dioxide include hydrochloric acid peptizer type anatase type titania sol (STS-02 manufactured by Ishihara Sangyo Co., Ltd. (average particle size 7 nm), ST-K01 manufactured by Ishihara Sangyo Co., Ltd.), nitric acid solution An anatase type titania sol (TA-15 manufactured by Nissan Chemical Co., Ltd. (average particle size 12 nm)) and the like can be mentioned.

  Further, a visible light responsive type may be used as the titanium oxide. Visible light responsive titanium oxide is also excited by the energy of visible light. Examples of such a visible light responsive method include a method of nitriding titanium oxide.

When titanium oxide (TiO ₂ ) is subjected to nitriding treatment, a new energy level is formed inside the band gap of titanium oxide (TiO ₂ ), and the band gap is narrowed. As a result, the excitation wavelength of titanium oxide (TiO ₂ ) is usually 380 nm, but it can be excited even by visible light having a longer wavelength than the excitation wavelength. As a result, the wavelength in the visible light region of energy irradiation from various light sources can also contribute to the excitation of titanium oxide (TiO ₂ ), so that it is possible to further increase the sensitivity of titanium oxide. .

Here, the nitriding treatment of titanium oxide as used in the present embodiment is a treatment for replacing part of the oxygen sites of the titanium oxide (TiO ₂ ) crystal with a nitrogen atom, or an interstitial of the titanium oxide (TiO ₂ ) crystal. Treatment of doping nitrogen atoms, or treatment of arranging nitrogen atoms at the grain boundaries of a polycrystalline aggregate of titanium oxide (TiO ₂ ) crystals.

The method of nitriding titanium oxide (TiO ₂ ) is not particularly limited. For example, crystalline titanium oxide fine particles are doped with nitrogen by heat treatment at 700 ° C. in an ammonia atmosphere, and the nitrogen is doped. Examples thereof include a method of forming a dispersion using fine particles and an inorganic binder, a solvent, or the like.

  Here, the smaller the particle size of the photocatalyst, the more effective the photocatalytic reaction occurs. The average particle size is preferably 50 nm or less, and it is particularly preferable to use a photocatalyst having a particle size of 20 nm or less.

  The photocatalyst-containing layer used in this step may be formed by the photocatalyst alone as described above, or may be formed by mixing with a binder.

  In the case of a photocatalyst-containing layer consisting only of a photocatalyst, the efficiency for decomposing and removing organic groups is improved, which is advantageous in terms of cost such as shortening of processing time. On the other hand, in the case of a photocatalyst-containing layer comprising a photocatalyst and a binder, there is an advantage that the formation of the photocatalyst-containing layer is easy.

  Examples of a method for forming a photocatalyst-containing layer composed only of a photocatalyst include a method using a vacuum film forming method such as a sputtering method, a CVD method, or a vacuum deposition method. By forming the photocatalyst-containing layer by vacuum film-forming method, it is possible to obtain a photocatalyst-containing layer that is a uniform film and contains only the photocatalyst, which enables uniform decomposition and removal of organic groups, etc. It is.

  Examples of a method for forming a photocatalyst-containing layer composed only of a photocatalyst include a method in which amorphous titania is formed on a substrate when the photocatalyst is titanium dioxide, and then the phase is changed to crystalline titania by firing. As the amorphous titania used here, for example, hydrolysis, dehydration condensation, tetraethoxytitanium, tetraisopropoxytitanium, tetra-n-propoxytitanium, tetrabutoxytitanium, titanium inorganic salts such as titanium tetrachloride and titanium sulfate, An organic titanium compound such as tetramethoxytitanium can be obtained by hydrolysis and dehydration condensation in the presence of an acid. Next, it can be modified to anatase titania by baking at 400 ° C. to 500 ° C. and modified to rutile type titania by baking at 600 ° C. to 700 ° C.

  Moreover, when using a binder, what has the high bond energy that the main frame | skeleton of a binder is not decomposed | disassembled by photoexcitation of said photocatalyst is preferable, for example, organopolysiloxane etc. can be mentioned.

  When organopolysiloxane is used as a binder in this way, the photocatalyst-containing layer is prepared by dispersing the photocatalyst and the binder organopolysiloxane in a solvent together with other additives as necessary. The coating solution can be formed by coating on a substrate. As the solvent to be used, alcohol-based organic solvents such as ethanol and isopropanol are preferable. The application can be performed by a known application method such as spin coating, spray coating, dip coating, roll coating, or bead coating. When an ultraviolet curable component is contained as the binder, the photocatalyst-containing layer can be formed by irradiating ultraviolet rays and performing a curing treatment.

An amorphous silica precursor can be used as the binder. This amorphous silica precursor is represented by the general formula SiX _4, X is a halogen, a methoxy group, an ethoxy group or a silicon compound an acetyl group or the like, and silanol or average molecular weight of 3,000 or less, their hydrolysates Polysiloxane is preferred.

  Specific examples include tetraethoxysilane, tetraisopropoxysilane, tetra-n-propoxysilane, tetrabutoxysilane, and tetramethoxysilane. In this case, the amorphous silica precursor and the photocatalyst particles are uniformly dispersed in a non-aqueous solvent, hydrolyzed with moisture in the air on the substrate to form silanol, and then at room temperature. A photocatalyst-containing layer can be formed by dehydration condensation polymerization. If dehydration condensation polymerization of silanol is carried out at 100 ° C. or higher, the degree of polymerization of silanol increases and the strength of the film surface can be improved. Moreover, these binders can be used individually or in mixture of 2 or more types.

  When the binder is used, the content of the photocatalyst in the photocatalyst containing layer can be set in the range of 5 to 60% by weight, preferably 20 to 40% by weight. The thickness of the photocatalyst containing layer is preferably in the range of 0.05 to 10 μm.

  In addition to the photocatalyst and the binder, the photocatalyst containing layer can contain a surfactant. Specifically, hydrocarbons such as NIKKOL BL, BC, BO, BB series manufactured by Nikko Chemicals Co., Ltd., ZONYL FSN, FSO manufactured by DuPont, Surflon S-141, 145 manufactured by Asahi Glass Co., Ltd., Dainippon Megafac F-141, 144 manufactured by Ink Chemical Industry Co., Ltd., Footgent F-200, F251 manufactured by Neos Co., Ltd., Unidyne DS-401, 402 manufactured by Daikin Industries, Ltd., Fluorard FC-170 manufactured by 3M Co., Ltd. Fluorine-based or silicone-based nonionic surfactants such as 176, and cationic surfactants, anionic surfactants, and amphoteric surfactants can also be used.

  In addition to the above surfactants, the photocatalyst-containing layer includes polyvinyl alcohol, unsaturated polyester, acrylic resin, polyethylene, diallyl phthalate, ethylene propylene diene monomer, epoxy resin, phenol resin, polyurethane, melamine resin, polycarbonate, Polyvinyl chloride, polyamide, polyimide, styrene butadiene rubber, chloroprene rubber, polypropylene, polybutylene, polystyrene, polyvinyl acetate, polyester, polybutadiene, polybenzimidazole, polyacrylonitrile, epichlorohydrin, polysulfide, polyisoprene, oligomers, polymers, etc. It can be included.

(2) Base The photocatalyst-containing layer side substrate used in this step has at least a base 12 and a photocatalyst-containing layer 11 formed on the base 12 as shown in FIG.
At this time, the material constituting the substrate to be used is appropriately selected depending on the energy irradiation direction, which will be described later, and whether the microarray chip has transparency.

  That is, for example, when the microarray chip is opaque, the energy irradiation direction is necessarily from the photocatalyst containing layer side substrate side. For example, as shown in FIG. It is necessary to irradiate with energy. As will be described later, the photocatalyst-containing layer side light-shielding portion is formed in advance in a predetermined pattern on the photocatalyst-containing layer side substrate, and the photocatalyst-containing layer side light-shielding portion is used to form a pattern. It is necessary to irradiate energy from the side substrate side. In such a case, the substrate needs to be transparent.

  On the other hand, if the microarray chip is transparent, a photomask can be arranged on the microarray chip side to irradiate energy. In such a case, the transparency of the substrate is not particularly required.

  Such a substrate may be flexible, such as a resin film, or may not be flexible, such as a glass substrate. This is appropriately selected depending on the energy irradiation method.

As described above, the material used for the substrate used for the photocatalyst-containing layer side substrate is not particularly limited, but in this step, the photocatalyst-containing layer side substrate is used repeatedly, so that the predetermined substrate is used. A material having strength and having a surface with good adhesion to the photocatalyst-containing layer is preferably used.
Specific examples include glass, ceramic, metal, and plastic.

  In order to improve the adhesion between the substrate surface and the photocatalyst containing layer, an anchor layer may be formed on the substrate. Examples of such an anchor layer include silane-based and titanium-based coupling agents.

(3) Photocatalyst containing layer side light-shielding part As the photocatalyst containing layer side light shielding part used for this process, you may use what the photocatalyst containing layer side light shielding part formed in pattern shape was formed. By using the photocatalyst containing layer side substrate having the photocatalyst containing layer side light shielding portion in this way, it is not necessary to use a photomask or perform drawing irradiation with a laser beam at the time of exposure. Therefore, since alignment between the photocatalyst-containing layer side substrate and the photomask is not necessary, it is possible to use a simple process, and an expensive apparatus necessary for drawing irradiation is also unnecessary, so that the cost is reduced. Has the advantage of being advantageous.

  The photocatalyst-containing layer side substrate having such a photocatalyst-containing layer side light-shielding part can have the following two modes depending on the formation position of the photocatalyst-containing layer side light-shielding part.

  For example, as shown in FIG. 4, a photocatalyst containing layer side light shielding portion 16 is formed on a substrate 12, and a photocatalyst containing layer 11 is formed on the photocatalyst containing layer side light shielding portion 16 to form a photocatalyst containing layer side substrate. It is an aspect to make. For example, as shown in FIG. 5, a photocatalyst containing layer 11 is formed on a substrate 12, and a photocatalyst containing layer side light shielding portion 16 is formed thereon to form a photocatalyst containing layer side substrate.

  In any embodiment, the photocatalyst-containing layer-side light-shielding portion is disposed in the vicinity of the portion where the photocatalyst-containing layer and the specific biopolymer etc. are located with a gap as compared with the case where a photomask is used. Therefore, since the influence of energy scattering in the substrate or the like can be reduced, the energy pattern irradiation can be performed very accurately.

  Furthermore, in the embodiment in which the photocatalyst containing layer side light shielding portion is formed on the photocatalyst containing layer, when the photocatalyst containing layer and the specific biopolymer are arranged with a predetermined gap, the photocatalyst containing layer side By making the film thickness of the light shielding portion coincide with the width of the gap, there is an advantage that the photocatalyst containing layer side light shielding portion can be used as a spacer for making the gap constant.

  That is, when the photocatalyst-containing layer and the specific biopolymer etc. are arranged in contact with each other with a predetermined gap, the photocatalyst-containing layer side light shielding part and the specific biopolymer etc. are brought into close contact with each other. By arranging in a state, it becomes possible to make the predetermined gap accurate, and by irradiating energy from the photocatalyst-containing layer side substrate in this state, it is possible to decompose and remove the organic group with high accuracy. It becomes.

  The method for forming the photocatalyst-containing layer side light-shielding part used in this step is not particularly limited, and is appropriately determined according to the characteristics of the formation surface of the photocatalyst-containing layer side light-shielding part, the shielding property against the required energy, and the like. Selected and used.

  For example, it may be formed by forming a metal thin film of chromium or the like having a thickness of about 1000 to 2000 mm by a sputtering method, a vacuum deposition method, or the like, and patterning the thin film. As this patterning method, a normal patterning method such as sputtering can be used.

  Alternatively, a method may be used in which a layer containing light-shielding particles such as carbon fine particles, metal oxides, inorganic pigments, and organic pigments in a resin binder is formed in a pattern. As the resin binder to be used, polyimide resin, acrylic resin, epoxy resin, polyacrylamide, polyvinyl alcohol, gelatin, casein, cellulose, or a mixture of one or more kinds, photosensitive resin, or O / A W emulsion type resin composition, for example, an emulsion of a reactive silicone can be used. The thickness of the resin photocatalyst-containing layer side light-shielding part can be set within a range of 0.5 to 10 μm. As a method for patterning the light shielding part on the resin photocatalyst-containing layer side, a commonly used method such as a photolithography method or a printing method can be used.

  In the above description, the two positions of the photocatalyst containing layer side light shielding portion between the substrate and the photocatalyst containing layer and the surface of the photocatalyst containing layer have been described as the formation position of the photocatalyst containing layer side. It is also possible to adopt a mode in which the photocatalyst-containing layer side light shielding portion is formed on the surface that is not provided. In this aspect, for example, a case where the photomask is brought into close contact with the surface so as to be detachable can be considered, and it can be suitably used when the energy irradiation region is changed in a small lot.

(4) Primer layer Next, the primer layer used for a photocatalyst containing layer side substrate is explained. In this step, when the photocatalyst-containing layer side light-shielding portion is formed in a pattern on the substrate as described above, and the photocatalyst-containing layer is formed thereon to form a photocatalyst-containing layer side substrate, the photocatalyst-containing layer A primer layer may be formed between the side light shielding part and the photocatalyst containing layer.

  The action and function of this primer layer are not always clear, but by forming a primer layer between the photocatalyst containing layer side light shielding part and the photocatalyst containing layer, the primer layer decomposes and removes organic groups by the action of the photocatalyst. Impurities from the openings present between the photocatalyst containing layer side light shielding part and the photocatalyst containing layer side light shielding part, which are factors that hinder, etc. It is considered that it has a function of preventing diffusion of impurities such as ions. Therefore, by forming the primer layer, processing such as decomposition and removal of the organic group proceeds with high sensitivity, and as a result, the organic group in the space portion can be removed with a high resolution pattern.

  In addition, since the said primer layer prevents that the impurity which exists in the opening formed between not only the photocatalyst containing layer side light shielding part but between the photocatalyst containing layer side light shielding parts influences the effect | action of a photocatalyst, primer The layer is preferably formed over the entire surface of the photocatalyst containing layer side light shielding part including the opening.

  The primer layer is not particularly limited as long as the primer layer is formed so that the photocatalyst containing layer side light-shielding portion of the photocatalyst containing layer side substrate is not in contact with the photocatalyst containing layer.

The material constituting the primer layer is not particularly limited, but an inorganic material that is not easily decomposed by the action of the photocatalyst is preferable. Specific examples include amorphous silica. When such amorphous silica is used, the precursor of the amorphous silica is represented by the general formula SiX ₄ and X is a silicon compound such as halogen, methoxy group, ethoxy group, or acetyl group, Silanol which is a hydrolyzate thereof or polysiloxane having an average molecular weight of 3000 or less is preferable.

  The thickness of the primer layer is preferably in the range of 0.001 μm to 1 μm, particularly preferably in the range of 0.001 μm to 0.1 μm.

b. Energy Irradiation Next, an energy irradiation method will be described. In this step, the photocatalyst-containing layer and the specific biopolymer or the immobilization layer are arranged with a gap between them, and the organic groups present in the space part are formed by irradiating energy from a predetermined direction. Can be removed.

  The above arrangement in this step means a state in which the action of the photocatalyst substantially extends to the organic group, and the photocatalyst-containing layer and the specific biopolymer are in close contact with each other. In addition, the photocatalyst-containing layer and the specific biopolymer are arranged at a predetermined interval. This gap is preferably 200 μm or less.

  In the present invention, the gap is preferably in the range of 0.2 μm to 10 μm, preferably in the range of 1 μm to 5 μm, considering that the photocatalyst is highly sensitive and the organic group is efficiently decomposed. preferable. Such a gap range is particularly effective for a small-area microarray chip that can control the gap with high accuracy.

  On the other hand, when processing a microarray chip having a large area of, for example, 300 mm × 300 mm or more, it is extremely difficult to form a fine gap as described above between the photocatalyst-containing layer side substrate and the specific biopolymer. Have difficulty. Therefore, when the microarray chip has a relatively large area, the gap is preferably in the range of 10 to 100 μm, particularly in the range of 50 to 75 μm. By setting the gap within such a range, there is no problem such as a decrease in accuracy of the pattern for removing the organic group or a problem that the sensitivity of the photocatalyst is deteriorated and the efficiency for decomposing and removing the organic group is deteriorated. Moreover, it is because there is an effect that unevenness does not occur in the removal of the organic group.

  Thus, when irradiating energy to a relatively large area microarray chip, the setting of the gap in the positioning device between the photocatalyst containing layer side substrate and the specific biopolymer in the energy irradiation device is within the range of 10 μm to 200 μm. In particular, it is preferable to set within the range of 25 μm to 75 μm. This is because by setting the set value within such a range, it is possible to arrange the photocatalyst without significantly deteriorating the sensitivity of the photocatalyst.

  Thus, by disposing the photocatalyst-containing layer and the specific biopolymer etc. at a predetermined interval, oxygen, water, and active oxygen species generated by the photocatalytic action are easily desorbed. That is, when the interval between the photocatalyst containing layer and the specific biopolymer is made narrower than the above range, it becomes difficult to desorb the active oxygen species, resulting in a slower rate of decomposition and removal of the organic group. If it is placed at a distance from the above range, it is difficult for the generated active oxygen species to reach the specific biopolymer, etc., and in this case also the rate of decomposition and removal of organic groups, etc. This is not preferable because there is a possibility of slowing down.

  As a method for uniformly forming such a very narrow gap and arranging the photocatalyst-containing layer and the specific biopolymer etc., for example, a method using a spacer can be mentioned. This is because a uniform gap can be formed by using the spacer in this way. In addition, by using such a spacer, the active oxygen species generated by the action of the photocatalyst reaches the surface of the specific biopolymer etc. at a high concentration without diffusing, so that the organic group is efficiently decomposed and removed. It can be performed.

  In this step, such a spacer may be formed as one member. However, for simplification of the process, as described in the column of the photocatalyst containing layer side substrate, the photocatalyst of the photocatalyst containing layer side substrate is used. It is preferable to form on the surface of the containing layer. In the above description of the photocatalyst-containing layer side substrate, the photocatalyst-containing layer side light-shielding portion has been described. However, in the present invention, such a spacer does not act on the surface of a specific biopolymer or the like. Since it only needs to have a function of protecting the surface, it may be made of a material that does not have a function of shielding the energy to be irradiated.

  In the case of using a photocatalyst-containing layer side substrate formed on a flexible substrate such as a resin film in which the photocatalyst-containing layer is flexible, it is difficult to provide the gap as described above. From the viewpoint of efficiency and the like, it is preferable that the photocatalyst-containing layer and the specific biopolymer are arranged so as to contact each other.

  In the present invention, such an arrangement state with a gap need only be maintained at least during energy irradiation.

  Next, energy irradiation is performed in a state where the arrangement as described above is maintained. The energy irradiation (exposure) as used in the present invention is a concept including irradiation of any energy beam capable of decomposing and removing an organic group by the photocatalyst containing layer, and is limited to visible light irradiation. is not.

  Usually, the wavelength of light used for such exposure is set in the range of 400 nm or less, preferably in the range of 380 nm or less. This is because, as described above, the preferred photocatalyst used in the photocatalyst-containing layer is titanium dioxide, and light having the above-described wavelength is preferable as the energy for activating the photocatalytic action by the titanium dioxide.

  Examples of light sources that can be used for such exposure include mercury lamps, metal halide lamps, xenon lamps, excimer lamps, and various other light sources.

  In addition to the method of performing pattern irradiation through a photomask using the light source as described above, it is also possible to use a method of drawing and irradiating in a pattern using a laser such as excimer or YAG. In this case, there is an advantage that a photomask is not necessary.

In addition, the amount of energy applied upon exposure is set to an amount necessary to decompose and remove the organic groups present in the space portion by the action of the photocatalyst in the photocatalyst-containing layer.
At this time, it is preferable in that the photocatalyst-containing layer is exposed while being heated, so that the sensitivity can be increased and the characteristic can be changed efficiently. Specifically, it is preferable to heat within a range of 30 ° C to 80 ° C.

  The energy irradiation direction in this step is determined by whether the photocatalyst-containing layer side substrate or the manufactured microarray chip is transparent as described above.

  That is, when the photocatalyst containing layer side light-shielding portion is formed, it is necessary to perform exposure from the photocatalyst containing layer side substrate side, and in this case, the photocatalyst containing layer side substrate is transparent to the energy irradiated. There must be.

  In addition, the exposure direction when the photocatalyst-containing layer is formed in a pattern is irradiated from any direction as long as energy is applied to the portion where the photocatalyst-containing layer and the property change layer are in contact as described above. May be.

  Similarly, in the case of using the above-described spacer, irradiation may be performed from any direction as long as energy is applied to the contact portion.

  In the case of using a photomask, energy is irradiated from the side where the photomask is arranged. In this case, the substrate on the side where the photomask is arranged, that is, either the photocatalyst-containing layer side substrate or the microarray chip needs to be transparent.

  After the energy irradiation as described above is completed, the organic group in the space part can be removed by removing the photocatalyst-containing layer side substrate from the arrangement position.

(Other)
In addition, in the manufacturing method of the microarray chip | tip of this embodiment, you may have a required process suitably besides the process mentioned above. In this embodiment, as described above, the base material has a property change layer whose properties change due to the action of the photocatalyst accompanying energy irradiation, and the photocatalyst-containing layer of the photocatalyst-containing layer side substrate described above, It is preferable to have a characteristic changing step of changing the characteristic of the specific biopolymer immobilization part on the characteristic changing layer by irradiating energy after arranging the gap with the characteristic changing layer. Thereby, in the specific biopolymer immobilization step, the specific biopolymer-containing coating liquid can be attached along the pattern in which the characteristic of the characteristic change layer is changed, and the adjacent specific biopolymer is fixed. This is because the specific biopolymer can be easily fixed without contacting or mixing the contained coating liquid.

  In this embodiment, in addition to the property change layer, the base material may have an immobilization layer for immobilizing the specific biopolymer. This is because the specific biopolymer can be more firmly immobilized by the substrate having this immobilization layer, and a high-quality microarray chip can be obtained.

  Here, the property change layer and the immobilization layer may each be used alone. For example, the property change layer is changed to the pattern of the specific biopolymer immobilization portion by the property change process, An immobilization layer may be formed on the characteristic change layer whose characteristics have changed. This is because, by utilizing the advantages of the characteristic change layer and the immobilization layer, the specific biopolymer can be firmly immobilized in a high-definition pattern, and a higher quality microarray chip can be obtained. .

In addition, when the base material has such a characteristic change layer or an immobilization layer, the organic group of the characteristic change layer or the immobilization layer existing on the space portion may be removed in the organic group removal step. preferable. This is because, when such a characteristic change layer or an immobilization layer remains, these organic groups and the object to be inspected may adsorb unspecifically.
Here, the characteristic change layer and the immobilization layer are usually formed on a support, and as the support, those used as the above-described base material can be used.
Hereinafter, the characteristic changing step as described above and the immobilization layer used in this embodiment will be described.

a. Characteristic Change Process First, the characteristic change process in this embodiment will be described. In the characteristic changing step in this embodiment, the base material has a characteristic changing layer whose characteristics change due to the action of the photocatalyst accompanying energy irradiation, and the photocatalyst containing layer on the photocatalyst containing layer side substrate described above is used as the characteristic changing layer. And a gap, and then, by irradiating energy, the characteristic of the specific biopolymer immobilization part on the characteristic change layer is changed.

  The base material used in this step is one in which the base material 1 has a support 5 and a characteristic change layer 6 as shown in FIG. In this step, for example, the photocatalyst-containing layer 11 of the photocatalyst-containing layer side substrate 13 having the photocatalyst-containing layer 11 and the substrate 12 is disposed so as to face the characteristic change layer 6, and a specific biological body is used using, for example, a photomask 14 or the like. The pattern 15 of the polymer immobilization part a is irradiated with energy 15 (FIG. 6A) to form a pattern 6 ′ in which the characteristic of the specific biopolymer immobilization part a on the characteristic change layer 6 is changed (FIG. 6). (B)) step. In this embodiment, the specific biopolymer-containing coating solution is attached by using the difference in the characteristics of the property change layer by performing the specific biopolymer immobilization step after the property change step. It can be made.

  Here, the photocatalyst containing layer side substrate used in this step, the irradiated energy, the arrangement of the photocatalyst containing layer side substrate, and the like can be the same as those described in the above “organic group removing step”. Therefore, detailed description here is omitted, and the characteristic change layer used in this step will be described below.

The property changing layer used in this step is a layer that changes due to the action of a photocatalyst accompanying energy irradiation, and a layer on which the specific biopolymer-containing coating solution can be easily attached using this property. However, in this step, the property change layer is a wettability change layer in which a pattern in which the wettability is changed by the action of the photocatalyst is formed, and the property change layer is decomposed by the action of the photocatalyst. The two cases of the decomposition removal layer from which the pattern due to the unevenness is formed are preferable because they bring out the effectiveness of the present invention.
Hereinafter, the wettability changing layer and the decomposition removal layer will be described.

(1) Wettability change layer Although the wettability change layer used for the base material of this embodiment is not particularly limited as long as the surface wettability is changed by the action of the photocatalyst accompanying energy irradiation, In particular, the layer is preferably a layer whose wettability changes so that the contact angle with water on the wettability changing layer surface is lowered by the action of the photocatalyst accompanying the irradiation of energy.

  In this way, when the wettability changing layer whose wettability changes so that the contact angle with water is reduced by energy irradiation, the above-mentioned photocatalyst-containing layer side substrate faces the wettability changing layer, and the energy pattern By performing irradiation, the wettability can be easily changed into a pattern. As a result, a pattern composed of a lyophilic region having a small contact angle with water and a liquid repellent region having a large contact angle with water is formed on the wettability changing layer. Here, the lyophilic region is a region having a small contact angle with water, and refers to a region having good wettability with respect to the specific biopolymer-containing coating solution and the like. The liquid repellent region is a region having a large contact angle with water, and refers to a region having poor wettability with respect to a specific biopolymer-containing coating solution. In the present embodiment, the region is referred to as a lyophilic region if the contact angle with water is 1 ° or more smaller than the adjacent region, and conversely, the region is in contact with water than the adjacent region. If the angle is larger than 1 °, it is defined as a liquid repellent region.

  The wettability changing layer used in this step is a wettability changing layer in which the contact angle with water in the part not irradiated with energy is a contact angle that is 1 ° or more larger than the contact angle with water in the part irradiated with energy. It is preferable that the angle be 5 ° or more, particularly 10 ° or more.

  If the difference between the contact angle with water in the part not irradiated with energy and the contact angle with water in the part irradiated with energy is less than the specified range, use the difference in wettability to determine the specificity. This is because it is difficult to attach the biopolymer-containing coating solution to the specific biopolymer fixing portion.

  As a specific contact angle with water in such a wettability changing layer, a contact angle with water in a portion not irradiated with energy is 30 ° or more, particularly 40 ° or more, particularly 50 ° or more. preferable. This is because the part that has not been irradiated with energy is a part that requires liquid repellency in this step, so when the contact angle with water is small, the liquid repellency is not sufficient, and the specific biopolymer This is because the contained coating liquid may spread to unnecessary portions and the adjacent specific biopolymers may come into contact with or mix with each other.

  On the other hand, in the case of energy irradiation, the wettability with water is preferably good, and specifically, the contact angle with water is 20 ° or less, particularly 10 ° or less. preferable. If the contact angle with water in the part irradiated with energy is high, the spread of the specific biopolymer-containing coating liquid in this part may be inferior, and problems such as lack of the specific biopolymer may occur. Because there is sex.

  In addition, the contact angle with water here is measured using a contact angle measuring instrument (Kyowa Interface Science Co., Ltd. CA-Z type) with water or a test liquid having the same surface tension. 30 seconds after dropping water droplets from the microsyringe), the result was obtained.

  Further, when the wettability changing layer as described above is used in the present embodiment, fluorine is contained in the wettability changing layer, and the fluorine content on the surface of the wettability changing layer is more than the wettability changing layer. The wettability changing layer may be formed so that when energy is irradiated, the wettability changing layer is lowered by the action of the photocatalyst as compared with before energy irradiation.

  In the microarray chip having such a feature, a pattern composed of a portion having a small fluorine content can be easily formed by pattern irradiation with energy. Here, fluorine has an extremely low surface energy. Therefore, the surface of a substance containing a large amount of fluorine has a smaller critical surface tension. Therefore, the critical surface tension of the portion having a small fluorine content is larger than the critical surface tension of the surface of the portion having a large fluorine content. This means that the portion with a low fluorine content is a lyophilic region compared to the portion with a high fluorine content. Therefore, forming a pattern composed of a portion having a lower fluorine content than the surrounding surface forms a pattern of a lyophilic region in the liquid repellent region.

  When such a wettability changing layer is used, the photocatalyst-containing layer and the wettability changing layer are arranged so as to face each other, and the pattern of the lyophilic region is easily formed in the liquid repellent region by irradiating the pattern with energy. This is because the specific biopolymer-containing coating solution can be attached only to the lyophilic region, and the specific biopolymer can be more easily immobilized.

  The fluorine content in the lyophilic region having a low fluorine content formed by irradiation with energy is 10 or less, preferably 5 or less, particularly preferably when the fluorine content of the portion not irradiated with energy is 100. 1 or less.

  By setting it within such a range, it is possible to make a large difference in lyophilicity between the energy-irradiated portion and the unirradiated portion. Therefore, by attaching a specific biopolymer-containing coating solution to such a wettability changing layer, it is possible to accurately fix the specific biopolymer only in the lyophilic region having a reduced fluorine content. This is because a microarray chip can be obtained with high accuracy. This rate of decrease is based on weight.

  The fluorine content in such a wettability changing layer can be measured by various commonly used methods such as X-ray Photoelectron Spectroscopy, ESCA (Electron Spectroscopy for Chemical Analysis)), and any method capable of quantitatively measuring the amount of fluorine on the surface, such as X-ray fluorescence analysis and mass spectrometry.

  The material used for such a wettability changing layer is a material whose wettability changes due to the action of the photocatalyst in the photocatalyst-containing layer upon irradiation with energy, that is, the characteristics of the wettability changing layer described above. The main chain is not particularly limited as long as it has a main chain that is not easily deteriorated or decomposed by the action. For example, (1) chloro- or alkoxysilane is hydrolyzed and polycondensed by sol-gel reaction or the like to increase the strength. Examples thereof include organopolysiloxanes to be exhibited, and (2) organopolysiloxanes such as organopolysiloxanes crosslinked with reactive silicones having excellent liquid repellency and oil repellency.

In the case of (1) above, the general formula:
Y _n SiX _(4-n)
(Where Y is an alkyl group, fluoroalkyl group, vinyl group, amino group, phenyl group, chloroalkyl group, isocyanate group, or epoxy group, or an organic group containing these, and X is an alkoxyl group, acetyl group, or Represents halogen, n is an integer from 0 to 3)
It is preferable that it is the organopolysiloxane which is a 1 type, or 2 or more types of hydrolysis condensate or cohydrolysis condensate of the silicon compound shown by these. Here, the alkoxy group represented by X is preferably a methoxy group, an ethoxy group, a propoxy group, or a butoxy group. Moreover, it is preferable that the carbon number of the whole organic group shown by Y exists in the range of 1-20, especially in the range of 5-10.

  In particular, an organopolysiloxane containing a fluoroalkyl group can be preferably used. Specific examples include one or two or more hydrolyzed condensates and cohydrolyzed condensates of fluoroalkylsilanes. For example, those described in JP-A No. 2003-195029, which are known as fluorine-based silane coupling agents, can be used.

  Examples of the reactive silicone (2) include compounds having a skeleton represented by the following general formula.

However, n is an integer of 2 or more, R ^1, R ² are each a substituted or unsubstituted alkyl having 1 to 10 carbon atoms, alkenyl, aryl or cyanoalkyl group, the total molar ratio of 40% or less Vinyl, phenyl and phenyl halide. Further, those in which R ¹ and R ² are methyl groups are preferable because the surface energy becomes the smallest, and the methyl groups are preferably 60% or more by molar ratio. In addition, the chain end or side chain has at least one reactive group such as a hydroxyl group in the molecular chain.

  Moreover, you may mix the stable organosilicone compound which does not carry out a crosslinking reaction like dimethylpolysiloxane with said organopolysiloxane.

  The wettability changing layer used in this step can further contain a surfactant. Specifically, hydrocarbons such as NIKKOL BL, BC, BO, BB series manufactured by Nikko Chemicals Co., Ltd., ZONYL FSN, FSO manufactured by DuPont, Surflon S-141, 145 manufactured by Asahi Glass Co., Ltd., Dainippon Megafac F-141, 144 manufactured by Ink Chemical Industry Co., Ltd., Footgent F-200, F251 manufactured by Neos Co., Ltd., Unidyne DS-401, 402 manufactured by Daikin Industries, Ltd., Fluorard FC-170 manufactured by 3M Co., Ltd. Fluorine-based or silicone-based nonionic surfactants such as 176, and cationic surfactants, anionic surfactants, and amphoteric surfactants can also be used.

  In addition to the above surfactants, the wettability changing layer includes polyvinyl alcohol, unsaturated polyester, acrylic resin, polyethylene, diallyl phthalate, ethylene propylene diene monomer, epoxy resin, phenol resin, polyurethane, melamine resin, polycarbonate. Polyvinyl chloride, polyamide, polyimide, styrene butadiene rubber, chloroprene rubber, polypropylene, polybutylene, polystyrene, polyvinyl acetate, polyester, polybutadiene, polybenzimidazole, polyacrylonitrile, epichlorohydrin, polysulfide, polyisoprene, oligomers, polymers, etc. Can be contained.

  Such a wettability changing layer can be formed by preparing a coating liquid by dispersing the above-mentioned components in a solvent together with other additives as necessary, and coating the coating liquid on a support. it can. As the solvent to be used, alcohol-based organic solvents such as ethanol and isopropanol are preferable. The coating can be performed by a known coating method such as spin coating, spray coating, dip coating, roll coating or bead coating. In addition, in the case where an ultraviolet curable component is contained, the wettability changing layer can be formed by irradiating with ultraviolet rays and performing a curing treatment.

  The thickness of the wettability changing layer is preferably from 0.001 μm to 1 μm, particularly preferably in the range of 0.01 to 0.1 μm, from the relationship of the wettability change rate by the photocatalyst.

  The wettability changing layer used in this step is not particularly limited as long as the wettability changing layer is changed by the action of the photocatalyst as described above. preferable. If the photocatalyst is not contained in the wettability changing layer in this way, the semi-permanent action of the photocatalyst does not reach the microarray chip when used as a microarray chip, and can be used without problems for a long period of time. That's why.

The wettability changing layer is usually formed on a support, but when this wettability changing layer is a material having self-supporting property, the wettability changing layer itself is used as a base material. May be.
Examples of the material of the wettability changing layer having such a self-supporting property include polyethylene, polycarbonate, polypropylene, polystyrene, polyester, polyvinyl fluoride, acetal resin, nylon, ABS, PTFE, methacrylic resin, phenol resin, and polyfluoride. And vinylidene chloride, polyoxymethylene, polyvinyl alcohol, polyvinyl chloride, polyethylene terephthalate, and silicone.

  Here, when the wettability changing layer as described above is used in this embodiment, in the organic group removing step, the wettability changing layer in the space part may be completely removed, and the liquid repellency of the surface Only the organic group having silane may be removed, and the silane compound or the like may remain. In any case, the organic group on the surface is removed, and the object to be inspected can be prevented from non-specific adsorption or the like.

(2) Decomposition removal layer Next, the decomposition removal layer used for this process is demonstrated. The decomposition / removal layer used in this step is not particularly limited as long as the decomposition / removal layer in the portion irradiated with energy is decomposed and removed by the action of the photocatalyst accompanying energy irradiation.

  Such a decomposed / removed layer has a pattern consisting of a portion having a decomposed / removed layer and a portion having no decomposed / removed layer without performing a development process or a cleaning process, that is, unevenness, because the portion irradiated with energy is decomposed and removed by the action of a photocatalyst. A pattern can be formed. Thereby, the decomposition removal layer of the specific biopolymer immobilization part is decomposed and removed, and the specific biopolymer-containing coating solution or the like can be easily attached using the unevenness.

  This decomposition / removal layer is oxidatively decomposed and vaporized by the action of the photocatalyst by energy irradiation, and is therefore removed without any special post-treatment such as development / washing process. Depending on the material, a cleaning process or the like may be performed.

  Here, in this embodiment, it is preferable that the wettability of the decomposition / removal layer and the support or the like on which the decomposition / removal layer is formed are different. This is because the specific biopolymer-containing coating solution can be attached by utilizing the difference in wettability between the decomposition removal layer and the support as well as the unevenness.

  In this case, the contact angle of the decomposition / removal layer with respect to water is larger than the contact angle of the support with respect to water, and the contact angle of the decomposition / removal layer with respect to the liquid having the same surface tension as the water has It is preferably 1 ° or more, particularly 5 ° or more, and more preferably 10 ° or more.

  As a specific contact angle with water in such a decomposition / removal layer, the contact angle with water in a portion not irradiated with energy is preferably 30 ° or more, more preferably 40 ° or more, and particularly preferably 50 ° or more. . This is the portion that is not irradiated with energy and is required to have liquid repellency in this embodiment. Therefore, when the contact angle with water is small, the liquid repellency is not sufficient, and the specific biopolymer-containing coating solution may spread to unnecessary portions, so that the adjacent specific biomolecule This is because a polymer may be mixed.

  On the other hand, in the region where the support is exposed by energy irradiation, it is preferable that the wettability with water is good. Specifically, the contact angle with respect to water is 20 ° or less, particularly 10 ° or less. It is preferable that If the contact angle with water in the area exposed by energy irradiation is high, the spread of the coating solution containing the specific biopolymer in this part may be inferior, and there are problems such as lack of the specific biopolymer. This is because it may occur.

  In this case, the support may be surface-treated so that the surface is lyophilic. Examples of the surface treatment so that the surface of the material becomes lyophilic include lyophilic surface treatment by plasma treatment using argon or water, and the lyophilic layer formed on the support is Examples thereof include a silica film obtained by a sol-gel method of tetraethoxysilane. In addition, a contact angle with the specific biopolymer containing coating liquid here can be obtained by the method similar to the method mentioned above.

  Further, the thickness of the decomposition / removal layer in this embodiment is not particularly limited as long as it can be decomposed and removed by the action of the photocatalyst associated with the irradiated energy. Specifically, the thickness is preferably 0.001 μm to 1 μm, particularly preferably in the range of 0.01 to 0.1 μm.

  Specific examples of the film that can be used for the above-described decomposition removal layer include a film made of a fluorine-based or hydrocarbon-based resin having liquid repellency. These fluorine-based and hydrocarbon-based resins are not particularly limited as long as they have liquid repellency, and these resins are dissolved in a solvent, for example, a general composition such as a spin coating method. It can be formed by a film method.

  In this embodiment, a functional thin film, that is, a self-assembled monomolecular film, a Langmuir-Brocket film, an alternating adsorption film, and the like can be used to form a film without defects. It can be said that it is more preferable to use such a film forming method.

  Here, since the decomposition removal layer used for this process can use what was described in Unexamined-Japanese-Patent No. 2003-195029 etc., detailed description here is abbreviate | omitted.

  When the decomposition removal layer as described above is used in this embodiment, the decomposition removal layer remaining in the space portion is usually completely removed by performing the organic group removal step. It will be.

b. Next, the immobilization layer used in this embodiment will be described. The immobilization layer used in this embodiment is a layer for immobilizing a specific biopolymer. In this embodiment, when the base material has such an immobilization layer, the specific biopolymer can be firmly fixed on the specific biopolymer adhesion portion.

  As the immobilizing agent used in such an immobilizing layer, various agents can be used according to the immobilizing means for the specific biopolymer. As means for immobilizing a specific biopolymer, a well-known binding method can be used by those skilled in the art. For example, an electrostatic binding method, a covalent bond fixing method, or the like can be used. .

  In the case of the above-mentioned immobilization means by electrostatic binding, a solution in which a substance having a charge opposite to that of the specific biopolymer is dissolved as a fixing agent is prepared as an immobilization layer-forming coating solution. The immobilization layer is formed by coating on a support.

  For example, when the specific biopolymer is an oligonucleotide, since the oligonucleotide has an anionic property, a coating solution using a substance having a cationic property as a fixing agent is used. Then, this is coated and dried to form an immobilization layer.

In this case, examples of the cationic immobilizing agent to be used include poly L-lysine, poly (allylamine hydrochloride), poly (ethyleneimine), poly (diallyldimethylammonium chloride) and the like. L-lysine is preferred.

  On the other hand, as an immobilization method using a covalent bond (chemical bond), a compound having a functional group (for example, a carboxyl group, an amino group, or a hydroxyl group) is used as an immobilizing agent for an immobilizing agent forming coating. And a method of binding a specific biopolymer to this.

  Examples of the compound having such a functional group and having adhesiveness to the base material include compounds that adhere to the base material, such as a carbon-carbon bond, preferably by a siloxane bond. Examples of the compound having a siloxane bond with such a substrate include a compound having a trichlorosilyl group or a trialkoxysilyl group. The functional group is preferably a compound having amine, hydroxyl, thiol, carboxylic acid, ester, amide, epoxide, isocyanate, or isothiocyanate. Specific examples of such compounds include aminoalkyltrialkoxysilanes, aminoalkyltrichlorosilanes, hydroxyalkyltrialkoxy-silanes, hydroxyalkyltrichlorosilanes, carboxyalkyltrialkoxysilanes, polyethylene glycols, triethoxysilanes, epoxyalkyltrialkylsilanes. Examples include alkoxysilanes and combinations thereof.

  As the method for forming the immobilization layer, a solution containing the silane coupling agent is used, and applied to the entire surface by a dip coating method, a transfer method, a spin coating method, or the like, and a silane coupling agent is attached, an inkjet, a dispenser Etc., and a method of applying only the hydrophilic region can be mentioned. A microarray chip can be formed by adhering a specific biopolymer on the immobilization layer thus formed by the specific biopolymer immobilization step.

  In this embodiment, among the above, a silane coupling agent containing an amino group is preferably used. In this case, the amino group of the silane coupling agent, which is the immobilizing agent, and a biopolymer whose end is modified with an amino group, specifically a terminal amino group such as an oligonucleotide, are covalently bonded to form a firm membrane. Can be immobilized.

  In this case, as the silane coupling agent having an amino group to be used, N-β (aminoethyl) γ-aminopropyltrimethoxysilane, N-β (aminoethyl) γ-aminopropylmethyldimethoxysilane, γ-aminopropyl is used. Examples include triethoxysilane, N-phenyl-γ-aminopropyltrimethoxysilane, and the like.

  Further, as another immobilization method by covalent bond between the immobilization layer and the specific biopolymer, for example, when the biopolymer is an oligonucleotide, a method of covalently bonding the 5 ′ end to the functional group It has been known. Specifically, first, the 5 ′ end of the oligonucleotide is reacted with a maleimide compound to introduce a maleimide group at the 5 ′ end of the oligonucleotide. Suitable maleimide compounds include sulfosuccinimidyl-4- (N-maleimidomethyl) cyclohexane-1-carboxylate and the like. Separately from the above reaction, the immobilization agent having an amino group as a functional group and succinimidyl-S-acetylthioacetate are reacted, and then deacetylation is performed using hydroxyramine, whereby the immobilization agent is converted into SH. Giving a group. By reacting the SH group on the immobilizing agent thus provided with the maleimide group introduced at the 5 ′ end of the oligonucleotide prepared by introducing a maleimide group at the 5 ′ end, the oligonucleotide and the immobilizing agent are reacted. The specific biopolymer can be immobilized on the immobilization layer.

  In addition to the above-described methods, many immobilization methods such as an immobilization layer using a biotin-avidin system can also be used.

  As will be described later, it is preferable that the immobilization layer used in this embodiment does not spread from the immobilization layer when a specific biopolymer-containing coating solution is adhered thereon. Therefore, in the present embodiment, in particular, the base material has a characteristic change layer and an immobilization layer, and the immobilization layer is formed along a pattern in which the characteristic of the characteristic change layer is changed. It is preferred that a specific biopolymer is deposited on the layer.

  Here, when the above-described immobilization layer is used in this embodiment, the immobilization layer in the space portion may be completely removed when the organic group removal step is performed, and the immobilization layer may be immobilized. Part of the layer may remain. For example, when the immobilization layer has a portion made of an inorganic material such as a silane compound bonded to the base material, the portion made of the inorganic material may be removed simultaneously with the organic group, and the portion made of the inorganic material It may remain on the substrate surface.

2. Second Embodiment Next, a second embodiment of the microarray chip of the present invention will be described. The method for producing a microarray chip of this embodiment includes a specific biopolymer immobilization step of immobilizing a specific biopolymer on a substrate,
And a defective portion removal step of removing the specific biopolymer in the defective portion when a defect occurs in the specific biopolymer immobilization step.

  In the microarray chip manufacturing method of this embodiment, for example, as shown in FIG. 7, the specific biopolymer-containing coating solution is attached to the specific biopolymer fixing portion a on the substrate 1 and solidified. The specific biopolymer 2 (2 ′, 2 ″, 2 ″ ″, etc.) is immobilized by a specific biopolymer immobilization step (FIG. 7A). In this embodiment, in the specific biopolymer immobilization step, for example, when the specific biopolymer 2 in the adjacent region is mixed and a defective portion 3 ′ is generated, or the target specific biopolymer is formed. When a defective portion 3 ″ formed with something different from the above occurs, a defective portion removing step (FIG. 7B) for removing the specific biopolymer 2 of the defective portion 3 is included.

  According to this embodiment, since the defective portion generated during the specific biopolymer immobilization step can be removed, in the manufactured microarray chip, for example, the defective portion and the test object are nonspecific. Can be prevented. In addition, when something different from the target specific biopolymer has been formed, only the defective portion is removed, and an immobilization layer is formed again on that portion to immobilize the specific biopolymer. It becomes possible. Thereby, a microarray chip without defects can be manufactured.

  Here, the defective portion removal step may be performed after the specific biopolymer immobilization step, or may be performed during the specific biopolymer immobilization step. Here, in this embodiment, for example, when a defective part occurs in a part of the target specific biopolymer immobilization part, if the defective part is removed, there is no problem when a microarray chip is obtained. In this case, it is only necessary to perform a defective portion removing process for removing only the defective portion. However, for example, the defective portion occupies most of the specific biopolymer fixing portion, and the specific biopolymer of the portion is lacking. Therefore, in the case where trouble is caused when the microarray chip is used, after performing the defective portion removing step for removing the defective portion, the defective portion removing portion from which the defective portion is removed, for example, as shown in FIG. It is preferable to perform the 2nd specific biopolymer immobilization process which fixes the target specific biopolymer 4 to c.

  In addition, it is preferable that the microarray chip manufactured in this embodiment also has the organic group removal step of the first embodiment described above. As a result, there is no organic group on the space part of the manufactured microarray chip, and the test object can be made a high-quality microarray chip without nonspecific adsorption or the like on the space part. is there.

  Also in this embodiment, the base material preferably has a characteristic change layer or an immobilization layer, and particularly preferably has a characteristic change layer forming step. Thereby, in the specific biopolymer immobilization step, the specific biopolymer-containing coating liquid can be attached along the pattern in which the characteristic of the characteristic change layer is changed, and the adjacent specific biopolymer is fixed. This is because the specific biopolymer can be easily fixed without contacting or mixing the contained coating liquid.

  Here, since the specific biopolymer immobilization step in this embodiment is the same as that in the first embodiment described above, description thereof will be omitted, and hereinafter, the defective portion removal step in this step, and the first step will be described. The bispecific biopolymer immobilization step will be described.

(Bad part removal process)
First, the defective part removing step in this step will be described. The defective portion removing step in this step is a step of removing the defective portion generated in the above-described specific biopolymer immobilization step. As described above, this step may be performed during the specific biopolymer immobilization step, or may be performed after the specific biopolymer immobilization step.

  On the other hand, when it is necessary to perform the second specific biopolymer immobilization step described later, for example, when the defective portion occupies most of the specific biopolymer immobilization portion, the defect has occurred. When it becomes clear, it can be said that it is preferable to perform a defective portion removing step with little loss.

  Here, what is removed by the defective portion removing step of the present embodiment is the specific biopolymer of the defective portion. For example, when the substrate has a property change layer or an immobilized layer, these The layer may be removed at the same time or may remain without being removed.

  The method for removing the defective portion in this step is not particularly limited as long as it is a method capable of removing the defective portion, and is performed by a commonly performed photolithography method or the like. For example, it may be one that irradiates ultraviolet light with a short wavelength in a pattern to remove the specific biopolymer and the like.

  In this embodiment, in particular, for example, as shown in FIG. 3, the photocatalyst containing layer 11 of the photocatalyst containing layer side substrate 13 having the photocatalyst containing layer 11 and the base 12 containing the photocatalyst is used as the specific biopolymer 3. It is preferable to perform the irradiation by irradiating energy 15 using, for example, a photomask 14 or the like after being arranged with a gap.

  This eliminates the need to use chemicals that adversely affect other parts of specific biopolymers, such as alkaline developers used in photolithography, for example. Moreover, by irradiating the defective portion with energy using the photocatalyst-containing layer side substrate, it is possible to easily decompose and remove organic groups, etc. This is because the molecule and the immobilization layer can be removed.

  Here, the photocatalyst-containing layer side substrate used in the present embodiment and the energy to be irradiated are the same as those used in the organic group removal step of the first embodiment described above, and thus the description thereof is omitted here. To do.

(Second specific biopolymer immobilization step)
Next, the second specific biopolymer immobilization step in this embodiment will be described. In the second specific biopolymer immobilization step in the present embodiment, the specific biopolymer immobilization step is immobilized in the specific biopolymer immobilization step on the defective portion removal portion from which the defective portion has been removed by the defective portion removal step. This is a step of immobilizing a specific biopolymer similar to a molecule.

  Examples of the method for immobilizing the specific biopolymer in this step include a method of adhering and solidifying the specific biopolymer-containing coating solution, which is the same as the specific biopolymer immobilization step. Detailed explanation here is omitted.

B. Next, the microarray chip of the present invention will be described. The microarray chip of the present invention has a substrate and a specific biopolymer fixed on the substrate in a pattern.

  For example, as shown in FIG. 9, the microarray chip of the present invention has a base material 1 and a specific biopolymer 2 formed in a pattern on the base material 1. Thereby, it can be set as the high quality microarray chip | tip which does not carry out nonspecific adsorption | suction etc. of a to-be-tested object etc. to the space part b which is an area | region between the adjacent specific biopolymers 2. FIG.

  Here, as shown in FIG. 10, for example, the microarray chip of the present invention includes a base material 1, a characteristic change layer 6 whose characteristics are changed by the action of a photocatalyst accompanying energy irradiation on the base material 1, and its characteristic changes. It may have the specific biopolymer 2 formed on the layer 6.

  As used herein, the property change layer may be a layer before the property changes as long as the property changes due to the action of the photocatalyst associated with energy irradiation, or a layer after the property change. There may be. For example, when a layer whose wettability changes due to the action of a photocatalyst associated with energy irradiation is used as the characteristic change layer, the specific biopolymer is usually formed on the wettability change layer after this wettability has changed. It will be. In this case, a wettability changing layer having changed wettability may be formed in the space between the fixed specific biopolymers. For example, when a layer containing an organopolysiloxane is used as the wettability changing layer, a layer made of polysiloxane having a hydroxyl group at the terminal is formed in the space portion by replacing the organic group of this organopolysiloxane with a hydroxyl group. May be. In this case, the organic group is removed from this portion, and therefore, a high-quality microarray chip can be obtained without non-specifically adsorbing the object to be inspected in this space portion. The layer made of polysiloxane having a hydroxyl group at the terminal is the removal of the organic group having liquid repellency of the organopolysiloxane used as the wettability changing layer described in the above-mentioned “A. Manufacturing method of microarray chip”. It shall be said to be a layer that has been made.

  Here, the substrate, the specific biopolymer, the property change layer, the formation method, and the like used in the microarray chip of the present invention are the same as those described in the above-mentioned “A. Microarray chip manufacturing method”. Therefore, explanation here is omitted.

  The present invention is not limited to the above embodiment. The above-described embodiment is an exemplification, and the present invention has substantially the same configuration as the technical idea described in the claims of the present invention, and any device that exhibits the same function and effect is the present invention. It is included in the technical scope of the invention.

  The following examples illustrate the present invention more specifically.

[Example 1]
(Preparation of photocatalyst-containing layer side substrate)
5 g of trimethoxymethylsilane (GE Toshiba Silicone Co., Ltd., TSL8113) and 2.5 g of 0.5 N hydrochloric acid were mixed and stirred for 8 hours. This was diluted 10 times with isopropyl alcohol to obtain an anchor composition.
The above anchor layer composition was applied onto a photomask substrate having a 250 μm pitch, 150 μm square opening by a spin coater, and dried at 150 ° C. for 10 minutes to obtain a transparent anchor layer (thickness 0.2 μm). ) Was formed.
Next, 30 g of isopropyl alcohol, 3 g of trimethoxymethylsilane (manufactured by GE Toshiba Silicone Co., Ltd., TSL8113) and 20 g of ST-K03 (manufactured by Ishihara Sangyo Co., Ltd.), which is a photocatalytic inorganic coating agent, are mixed at 100 ° C. Stir for 20 minutes. This was diluted 3 times with isopropyl alcohol to obtain a composition for a photocatalyst-containing layer.
A transparent photocatalyst-containing layer (thickness: 0.15 μm) is formed by applying the photocatalyst-containing layer composition on a photomask substrate on which an anchor layer is formed by a spin coater and performing a drying treatment at 150 ° C. for 10 minutes. Formed.

(Specific biopolymer formation)
Next, a fluoroalkylsilane dissolved in isopropyl alcohol was coated on the cleaned glass surface at 1,500 rpm for 3 minutes, and dried at 60 ° C. for 10 minutes. Then, after facing the photocatalyst containing layer on the photocatalyst containing layer side substrate with a gap of 50 μm, UV light with a wavelength of 255-350 nm was irradiated for 30 seconds with an illuminance of 20,000 μW / cm ² / nm and washed with water. -Dried.
Subsequently, when this substrate was dipped in a vat filled with pure water and pulled up with the pattern surface facing upward, the water remained only in the hydrophilic portion irradiated with energy.
A 3-aminopropyltrimethoxysilane solution dissolved in 95% ethanol in this glass substrate so as to be 1% is coated with a spinner at 1,500 rpm for 3 minutes, and is attached to the hydrophilic portion at 60 ° C. for 10 minutes. Dried. Thereafter, it was rinsed in a 2% glutaraldehyde aqueous solution for 5 hours, washed with water, and then dried at 110 ° C. for 30 minutes to form an immobilization layer.
Next, the above substrate was rinsed and coupled at 50 ° C. and 90% humidity for 5 hours using 1 μg of mouse nerve growth factor (NGF) gene dissolved in 10 mL of 50 mM Tris HCl at pH 7.0, and washed with water. , Dried. Furthermore, Cy fluorescent staining of this substrate was performed by a standard method, and DNA was fluorescently stained, washed with water and dried. At this time, fluorescence was observed in the hydrophilic portion, but thin fluorescence remained in the hydrophobic portion.
Using this photocatalyst containing layer side substrate similar to the above except that the light-shielding portions are 250 μm pitch and 100 μm □, this region is irradiated with energy in the same manner as described above, and cleaned. As a result of the drying, only the hydrophilic portion of 100 μm □ showed fluorescence, and the other space portions did not emit fluorescence.
The general reagent used in this example is manufactured by Wako Pure Chemical Industries, and the silane coupling agent is manufactured by Shin-Etsu Chemical. The DNA is manufactured by Nippon Gene Co., Ltd., and the fluorescent label is manufactured by Amersham.

[Example 2]
The above mouse nerve growth factor (NGF) gene application was performed in the same manner as in Example 1 except that dispensing with a pipette was performed. In this case, before the space portion was irradiated with energy, there was bleed around the hydrophilic portion, but due to the energy irradiation using the photocatalyst-containing layer side substrate, this bleed was not observed and only the hydrophilic portion of 100 μm □ showed fluorescence. Fluorescence disappeared in the space part.

It is process drawing which shows an example of the manufacturing method of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows an example of the characteristic change process in the manufacturing method of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows an example of the organic group removal process in the manufacturing method of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows an example of the photocatalyst containing layer side board | substrate used for the organic group removal process in the manufacturing method of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows the other example of the photocatalyst containing layer side board | substrate used for the organic group removal process in the manufacturing method of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows the other example of the photocatalyst containing layer side board | substrate used for the organic group removal process in the manufacturing method of the microarray chip | tip of this invention. It is process drawing which shows the other example of the manufacturing method of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows an example of the 2nd specific biopolymer immobilization process in the manufacturing method of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows an example of the microarray chip | tip of this invention. It is a schematic sectional drawing which shows the other example of the microarray chip | tip of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Base material 2 ... Specific biopolymer 3 ... Defect part 4 ... Specific biopolymer 11 ... Photocatalyst containing layer 12 ... Base | substrate 13 ... Photocatalyst containing layer side board | substrate 15 ... Energy

Claims (10)

A specific biopolymer immobilization step of immobilizing the specific biopolymer on the substrate;
And an organic group removing step of removing an organic group present in a space portion between the specific biopolymer immobilization portions. A specific biopolymer immobilization step of immobilizing the specific biopolymer on the substrate;
A method for producing a microarray chip, comprising: a defective portion removing step of removing a defective portion of the specific biopolymer when a defect occurs in the specific biopolymer immobilization step.   3. The second specific biopolymer immobilization step of immobilizing the specific biopolymer in a defective portion removal unit from which a defective portion has been removed by the defective portion removal step. Manufacturing method of microarray chip.   The base material has a characteristic change layer whose characteristics are changed by the action of a photocatalyst accompanying energy irradiation, the photocatalyst containing layer containing a photocatalyst and the photocatalyst containing layer of the photocatalyst containing layer side substrate having the substrate 2. A characteristic changing step of changing the characteristic of the specific biopolymer immobilization part on the characteristic changing layer by irradiating energy after being arranged with a gap from the changing layer. A method for manufacturing a microarray chip according to any one of claims 1 to 3.   5. The microarray chip according to claim 4, wherein the characteristic change layer is a wettability change layer in which wettability is changed so that a contact angle with water is lowered by an action of a photocatalyst accompanying energy irradiation. Method.   5. The method of manufacturing a microarray chip according to claim 4, wherein the characteristic change layer is a decomposition removal layer that is decomposed and removed by the action of a photocatalyst accompanying energy irradiation.   After the organic group removal step or the defective portion removal step, the photocatalyst-containing layer on the photocatalyst-containing layer side substrate having a photocatalyst-containing layer and a substrate is disposed with a gap from the specific biopolymer. The method of manufacturing a microarray chip according to claim 1, wherein the method is performed by irradiating energy.   A microarray chip comprising: a base material; and a specific biopolymer fixed in a pattern on the base material.   9. The microarray chip according to claim 8, wherein the specific biopolymer is formed on a property change layer whose properties are changed by the action of a photocatalyst accompanying energy irradiation formed on a substrate. 10. The microarray chip according to claim 9, wherein a layer made of polysiloxane having a hydroxyl group at a terminal is formed on a base material in a space portion which is a region between the specific biopolymers.

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