JP2009046624A - Solid-phase material for fixing microscopic object, solid-phase body with microscopic object fixed and method of manufacturing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a solid-phase material having high ability of selective fixation by photoirradiation, by suppressing the nonspecific adsorption of a microscopic object. <P>SOLUTION: The solid-phase material for photoimmobilization, for fixing the microscopic object by photoirradiation, includes a light responsive component causing the change of molecular structure or the change of molecular arrangement on a matrix of the solid-phase material by light, and an alkylene glycol residue group-containing component expressed by formula (1). <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a solid phase material for immobilizing a minute object, a solid phase body on which the minute object is immobilized, and a method for producing the same.

  As a technique for immobilizing a minute object, an optical immobilization method using light irradiation is known (Patent Document 1). The light immobilization method is a method of immobilizing a minute object by supplying a minute object to a solid phase surface containing a photoresponsive component and causing light deformation in the solid phase by light irradiation. By immobilizing minute objects such as proteins and DNA in a specific region on the solid phase by the light immobilization method, analysis and functional analysis of the immobilized minute objects and substances that interact with the minute objects are performed on the solid phase. Utilization as a protein chip or a DNA chip is under study. In order to increase the detection accuracy and accuracy in the interaction and reaction on the surface of such a solid phase material, in addition to the adsorption of minute objects in the non-irradiated region, solid phase material can be generated during the production of the solid phase material. It is important to suppress adsorption of substances that react with immobilized micro objects and substances that are generated by the reaction (Patent Document 2).

  On the other hand, as one of the techniques for immobilizing a minute object on the surface of a solid phase material such as a substrate, a technique for immobilizing a minute object such as a protein on a solid phase by a covalent bond is known. As a material for suppressing adsorption of a minute object in a non-immobilized region in a covalent immobilization method or adsorption of a substance that reacts with the immobilized minute object, for example, an alkylene glycol residue and a minute object A polymer material having a functional group for immobilizing a substance is disclosed, and a minute object is immobilized on the surface of the material by a covalent bond between the functional group for immobilizing the minute object and the minute object (for example, patents) Reference 3).

  Moreover, a peptide array in which a substrate such as glass and a protein are bound in a form using a polyalkylene glycol as a linker is also disclosed (for example, Patent Document 4).

JP 2003-329682 A JP 2007-132866 A JP 2006-299045 A JP 2007-24729 A

  However, according to the present inventors, in detecting a highly accurate interaction with a minute object that is easily adsorbed, the adsorption suppressing ability of the solid phase material for light immobilization described in Patent Document 1 is not necessarily sufficient. However, it was found that a good S / N ratio could not be obtained.

  In addition, in order to use a solid-phase body on which a minute object is immobilized as a microreactor for performing an enzyme reaction or the like, a minute object is supplied non-selectively to a solid-phase material, and patterning during light irradiation is used. It is advantageous to immobilize minute objects. In order to improve the accuracy and miniaturization of minute object patterning, the difference between the amount of light immobilized on the light-irradiated surface and the amount of adsorption on the non-irradiated surface is reduced by sufficiently reducing the adsorption of minute objects on the non-irradiated surface. Need to be increased. That is, it is necessary to increase the contrast of the minute object between the light irradiation site and the light non-irradiation site.

  According to the present inventors, the solid phase material for light immobilization disclosed in the above-mentioned Patent Document 1 is insufficient for fine patterning, although the adsorption of minute objects on the light non-irradiated surface is relatively small. all right. Furthermore, in the method disclosed in Patent Document 2, the surface of the solid phase material other than the immobilization region is modified in order to immobilize the minute object in the minute region in an array, and the minute object is irradiated with light. It is not intended to be fixed by fine patterning. Furthermore, in order to use as a microreactor or the like, it is necessary to prevent adsorption of a substrate which is a reaction partner of the immobilized protein and a reaction product after the reaction over the entire reaction system on the surface of the solid phase material.

  On the other hand, in the covalent bond immobilization material disclosed in Patent Documents 3 and 4, the solid phase is hydrophilized by introducing polyethylene glycol or the like into the solid phase, whereby a micro object or the like in the non-immobilized region. It is trying to reduce the adsorption. However, the immobilization state by the light immobilization method is not a strong bond introduced from the outside like a covalent bond, but is due to an increase in the contact area between the micro object utilizing the optical deformation of the solid phase material and the solid phase material surface. As long as it is considered to be a thing, hydrophilizing the surface of the solid phase material that comes into contact with the minute object may cause an extreme decrease in immobilization ability. In addition, since polyethylene glycol is a molecule with high molecular mobility, there is also a possibility that the light deformation caused by light irradiation is restored and the immobilization ability is lowered.

  Furthermore, as the solid phase material, it is necessary to reduce the residual stress and increase the durability at the time of producing the solid phase material. However, the azo compound used for the light-fixing material is usually a rigid component, such as during molding. As a result, the film quality is likely to be non-uniform and there is a problem in durability. Such structural and mechanical properties may reduce the stability of the minute object.

  From the above, regarding the solid phase material used for the photo-immobilization method at present, the entire surface of the solid phase is fine while suppressing the adsorption of minute objects during non-light irradiation and the use of the solid phase material. No one has been able to achieve the light fixation of the object at the same time. In addition, no method for improving the mechanical properties of the light fixing material has been found.

  Therefore, the present invention provides a solid phase material that has improved the ability of selective immobilization by light irradiation by suppressing the adsorption of minute objects during non-irradiation and the use of the solid phase material, and a method for producing the same. Is one of the purposes. At the same time, another object is to provide a solid phase material excellent in structural and mechanical properties such as durability and homogeneity of the membrane and a method for producing the same. It is another object of the present invention to provide a solid phase material that can maintain the stability of a micro object once held on a solid phase and a method for producing the same.

According to the present invention, there is provided a solid phase material for light immobilization that immobilizes a minute object by light irradiation, and a photoresponsive component that causes a change in molecular structure or a molecular arrangement by light in a matrix of the solid phase material. And an alkylene glycol residue-containing component represented by the following formula (1).

However, R < ¹ > represents the hydrogen atom, a substituent, or the coupling group connected with respect to the structural component of the said matrix, and R < ² > represents a hydrogen atom or a C1-C10 alkyl group. Z represents an alkylene group having 1 to 10 carbon atoms, and n represents an integer of 1 to 100.

  Further, according to the present solid phase material, there is provided a solid phase material that immobilizes the minute object on the solid phase material by deforming the solid phase material for light immobilization by light irradiation.

  Furthermore, according to the present solid phase material, a solid phase material in which the amount of light immobilized by light irradiation is 3 times or more than the amount of adsorption when no light is irradiated is provided. Also provided is a solid phase material that is more durable against external stress than the same solid phase material except that it does not contain the alkylene glycol residue-containing component.

  Furthermore, according to the present solid phase material, a solid phase material in which the micro object fixed to the solid phase material has higher stability than the same solid phase material except that the component does not contain the alkylene glycol residue-containing component. Is provided.

  In the solid phase material, Z may represent an alkylene group having 1 to 4 carbon atoms. The n may be an integer of 4 to 25.

Furthermore, according to the present solid phase material, the matrix is a polymer, and an alkylene glycol residue represented by —O— (Z—O) n—R ² in the alkylene glycol residue-containing component with respect to the mass of the matrix. The solid phase material having a group content of 50% by mass or less is also provided. Alternatively, the content of the alkylene glycol residue may be 30% by mass or less.

In the present solid phase material, the matrix may be obtained by polymerizing a monomer composition containing a photoresponsive component-containing monomer and an alkylene glycol residue-containing monomer. At this time, the polyalkylene glycol residue-containing monomer may be represented by the following formula (2).

  However, Y represents a hydrogen atom or a methyl group.

According to the solid phase material, Z represents an alkylene group having 1 to 4 carbon atoms, the matrix is a polymer, and —O— (Z— in the alkylene glycol residue-containing component with respect to the mass of the matrix. O) The said solid-phase material whose content of the alkylene glycol residue represented by n-R < ² > is 30 mass% or less is also provided.

  In the solid phase material, the micro object may be a biomolecule. The minute object may be a protein.

  The solid phase material may be used for immobilization by selective light irradiation.

  According to the present invention, there is provided a solid phase body in which a micro object is immobilized, wherein the micro object is held on the surface of the solid phase material described above.

  Further, according to the present invention, there is provided a method for producing a solid phase in which a micro object is fixed, the preparation step for preparing any one of the above solid phase materials, and the surface of the solid phase material or the vicinity thereof And a holding step of holding the minute object in the solid phase by irradiating the minute object existing in the light.

  In the present manufacturing method, the holding step is a step of non-selectively supplying the micro object to the surface of the solid phase material and selectively irradiating a predetermined region of the surface of the solid phase material with light. Also good.

  According to the present invention, there is provided a method of using a solid body in which a micro object is immobilized, wherein a test sample is supplied to the micro object immobilized on the solid body, and the micro object and the test sample are There is provided a method of use comprising the steps of causing an interaction with a component of and a step of detecting the interaction.

  According to the solid phase material of the present invention, the alkylene glycol residue-containing component to be contained in the matrix of the solid phase material is expressed by the above formula (1), so that adsorption in a light non-irradiated region of a micro object and use of the solid phase material are performed. Adsorption occurring at the time can be effectively suppressed while maintaining the immobilization ability of the minute object. Usually, when polyethylene glycol is included in the matrix, it was thought that the immobilization ability was greatly reduced due to the hydrophilicity and molecular mobility of polyethylene glycol. What I could do was surprising. In addition, since it is possible to suppress adsorption during light non-irradiation and adsorption that occurs when using solid phase material over the entire surface of the solid phase, even if there are minute objects on the entire surface of the solid phase, A minute object can be reliably fixed only to the irradiated part. As a result, it is possible to increase the contrast of the minute object between the light irradiation part and the light non-irradiation part. Therefore, the solid phase material is suitable for immobilizing a minute object using fine patterning at the time of light irradiation.

  In addition, by containing a component with high molecular mobility such as an alkylene glycol residue, structural and mechanical properties such as uniformity and durability of the film quality are improved when the solid phase material is thinned. be able to.

  The solid phase of the present invention is obtained by immobilizing a micro object on the solid phase material of the present invention. As described above, this solid phase material has high ability to selectively immobilize a minute object by light irradiation because adsorption in the light non-irradiation region is suppressed while maintaining the ability to immobilize the minute object. That is, the contrast of the minute object is high between the light irradiation part and the light non-irradiation part. Therefore, this solid phase body is excellent in detection accuracy with respect to the interaction and reaction with a minute object. In addition, it is considered that the deterioration of minute substances, particularly biological substances, can be prevented by the presence of water molecules contained by the alkylene glycol residue component contained in the solid phase material on the surface of the minute objects. Moreover, the solid phase material has excellent structural and mechanical properties. Therefore, the solid phase body can stably maintain a micro object once held on the solid phase. In addition, according to the method for producing a solid phase of the present invention, not only the detection sensitivity to the interaction or reaction with a micro object is high, but also the micro object once held on the solid phase can be stably maintained. A solid phase can be obtained.

  As described above, the present invention relates to a light-immobilized solid phase material for immobilizing a minute object, a solid phase body on which the minute object is immobilized, a method for producing the same, and a method for using the same. Hereinafter, embodiments of the present invention will be described in detail.

(Solid phase material for light immobilization)
The solid phase material for light immobilization of the present invention is a material for immobilizing a minute object on the solid phase surface by light irradiation. The solid phase material for light immobilization of the present invention contains a photoresponsive component and an alkylene glycol residue-containing component represented by formula (1) in a matrix (matrix). Hereinafter, the solid phase material will be sequentially described. In addition, although the form shown below is a preferable form of this invention, it is not limited to this.

(matrix)
The matrix of the solid phase material is not particularly limited as long as it can hold the photoresponsive component and the alkylene glycol residue-containing component. For example, various organic materials including low molecular materials or high molecular materials, inorganic materials such as glass, organic-inorganic composite materials, and the like can be used. Considering the dispersibility and retention ability of the photoresponsive component and the alkylene glycol residue-containing component in the matrix, a polymer material or a composite material containing a polymer material is preferable.

  The polymer material constituting the matrix is not particularly limited, and various thermoplastic or thermosetting polymers can be used. For example, (1) polymers of carbon multiple bond monomers such as olefin polymers, vinyl polymers, acrylic polymers, methacrylic polymers, styrene polymers, diene polymers, and (2) cyclic monomers such as cyclic ether polymers. (3) polymers of bifunctional monomers such as ester polymers, urethane polymers, urea polymers, and the like. Among these, in consideration of the convenience of copolymerization, a polymer of a monomer having a double bond (hereinafter, also referred to as a double bond monomer) such as an olefin polymer, an acrylic polymer, or a methacrylic polymer, or a urethane type Polymers are preferred. More preferably, it is a double bond monomer. In particular, acrylic polymers, methacrylic polymers, and acrylic-methacrylic copolymers have little non-specific adsorption when immobilizing proteins such as antibodies, and therefore can effectively suppress background signals in antibody chips and microreactors. It is possible and preferable.

  It is preferable that the matrix is configured to cause shape deformation (hereinafter also referred to as light deformation) due to a change in the molecular structure of the photoresponsive component. In this specification, optical deformation includes deformation due to entanglement between a micro object and a solid-phase material surface due to movement at a molecular level, in addition to a change in shape in a normal macro sense. Some of these deformations cannot be clearly observed by ordinary observation means due to problems of deformation amount and deformation form. Photodeformation is considered to be induced by the presence of the photoresponsive component in the matrix material, for example, by changing the volume, density, free volume, etc. of the matrix material or photoresponsive component during light irradiation. It is done.

  The three-dimensional form of the solid phase material composed of the matrix is not particularly limited. In addition to a film-like body, a sheet-like body, and a plate-like body, various shapes such as a spherical body, an indefinitely shaped body, a needle-like body, a rod-like body, and a flake-like body can be adopted. The matrix may be held on a part of an appropriate support such as a substrate or particles or on the surface thereof. The support may be an inorganic material such as glass, silicon, or gold, or may be an organic material such as a polymer material. The thinning of the support can be performed by a known method such as spin coating, dip coating, and ink jet.

  In addition, components other than the polyalkylene glycol residue-containing component and the photoresponsive component may be added or bonded to the material constituting the matrix. For example, when the matrix material is a polymer material, it may contain a compatibilizing component for enhancing the compatibilization of the matrix material and the alkylene glycol residue-containing component. In order to include the compatibilizing component in the matrix, for example, a form in which a compatibilizing agent is added may be used, or a form in which the compatibilizing component is introduced into the alkylene glycol residue-containing component may be employed. Moreover, it does not specifically limit as a kind of compatibilizing component, A reactive type may be sufficient and a non-reactive type may be sufficient.

(Alkylene glycol residue-containing component)
The matrix contains an alkylene glycol residue-containing component represented by the above formula (1). Here, R ¹ in Formula (1) represents a hydrogen atom, a substituent, or a linking group connected to the constituent components of the matrix, and R ² is a hydrogen atom or an alkyl group having 1 to 10 carbon atoms. Represents. Z represents an alkylene group having 1 to 10 carbon atoms, and n represents an integer of 1 to 100.

R ¹ in formula (1) is a hydrogen atom, a substituent, or a linking group linked to a constituent component of the matrix. The substituent and the linking group are not particularly limited as long as they do not inhibit the light immobilization ability derived from the photoresponsive component or the nonspecific adsorption ability derived from the alkylene glycol residue. For example, when the alkylene glycol residue-containing component is dispersed in the matrix like an additive, the substituent may be a hydrogen atom or an alkyl group having 1 to 20 carbon atoms as in R ² , or a material constituting the matrix It is good also as a component and functional group in consideration of compatibility. The linking group may be, for example, a functional group that constitutes a part of the matrix, or a functional group that can be chemically bonded to the constituent components of the matrix. The linking group typically has a structure obtained as a result of polymerization of a polymerizable functional group of a monomer constituting the polymer when the matrix is a polymer.

R < ² > in Formula (1) is a hydrogen atom or a C1-C10 alkyl group. This is because when the number of carbon atoms exceeds 10, the hydrophilicity of the alkylene glycol residue-containing component is lowered, so that a desired suppression effect on adsorption that occurs when light is not irradiated or when a solid phase material is used cannot be obtained. . Preferably, they are hydrogen or a C1-C6 alkyl group, More preferably, they are a hydrogen atom or a C1-C4 alkyl group, More preferably, they are a hydrogen atom or a C1-C2 alkyl group. is there. Moreover, if it is a hydrogen atom or a C1-C4 alkyl group, it is preferable also from the point of availability.

  Z in Formula (1) is an alkylene group having 1 to 10 carbon atoms. When carbon number exceeds 10, the hydrophilic property of an alkylene glycol residue containing component will fall too much. An alkylene group having 1 to 6 carbon atoms is preferable for imparting hydrophilicity suitable for adsorbing a minute object at the time of non-irradiation to an alkylene glycol residue-containing component or suppressing adsorption occurring when using a solid phase material. More preferred are alkylene groups having 2 to 4 carbon atoms, and more preferred are alkylene groups having 2 carbon atoms. Moreover, if it is a C2-C4 alkylene group, it is preferable also from the point of availability.

  N in Formula (1) represents the number of repetitions of alkylene glycol (Z—O), and is an integer of 1 or more and 100 or less. If it exceeds 100, the influence of the alkylene glycol residue on the properties of the material constituting the matrix, that is, the molecular mobility of the alkylene glycol becomes too large, which may affect the durability of the matrix. Preferably, it is an integer of 1 or more and 50 or less, more preferably an integer of 2 or more and 30 or less, and further preferably an integer of 4 or more and 25 or less. If it is 4 or more and 25 or less, it is possible to reliably impart to the matrix the ability to suppress adsorption during light non-irradiation or the use of a solid phase material. Moreover, if it is an integer of 1-50, it is preferable also from the point of availability.

  In addition, the repeating number n may show the average repeating number of the mixture of the alkylene glycol residue containing component comprised by a different repeating number. In addition, when the repeating number n is 2 or more, the carbon number of the alkylene group in the repeated alkylene glycol (Z—O) may be the same or different.

  In addition, you may use the alkylene glycol residue containing component introduce | transduced into a matrix in combination of said 1 type, or 2 or more types.

(Content)
The content of the alkylene glycol residue-containing component is not particularly limited, and can be contained in an amount according to the size of the adsorption inhibiting ability to be imparted, the structural / mechanical properties, and the like. When the matrix material is a polymer, the content of the alkylene glycol residue represented by —O— (Z—O) n—R ² in the formula (1) is 1% by mass or more based on the mass of the matrix. It is preferable that it is 50 mass% or less. If it is less than 1% by mass, not only the effect of suppressing adsorption during non-irradiation cannot be sufficiently obtained, but also stress during molding tends to remain, so that the film quality is likely to be non-uniform and durability is poor. On the other hand, if it exceeds 50% by mass, the durability and water resistance of the matrix material may be lowered due to the large influence of alkylene glycol on the properties of the matrix material. More preferably, they are 5 mass% or more and 30 mass% or less. Within this range, a sufficient blending effect of the alkylene glycol residue-containing component can be obtained while ensuring durability and water resistance. This is because if it is in the range of more than 30% by mass and not more than 50% by mass, the effect of suppressing adsorption in the non-irradiated region can be obtained, but the amount of light immobilization may be slightly affected.

(Form containing an alkylene glycol residue-containing component)
The form in which the alkylene glycol residue-containing component is contained in the matrix is not particularly limited. For example, the alkylene glycol residue-containing component may be dispersed in the matrix as a separate component from the matrix material. Alternatively, it may be connected to one or more matrix materials constituting the matrix via a chemical bond such as a covalent bond to be a part thereof. Preferably, the matrix material is a polymer material, and an alkylene glycol residue-containing component is dispersed or linked in the polymer material. More preferably, it is an embodiment in which the polymer material is linked to a constituent component. By doing so, it is possible to prevent the alkylene glycol residue-containing component from flowing out of the matrix when a liquid medium is used in a processing step such as light immobilization. In addition, since the dispersibility of the alkylene glycol residue-containing component in the matrix material is improved, it is possible to uniformly impart to the surface of the matrix the ability to suppress adsorption during light non-irradiation or use of a solid phase material. . As a result, it is possible to achieve suppression over adsorption over the entire surface of the matrix when adsorption is not performed or when solid phase materials are used.

  In order to introduce an alkylene glycol residue-containing component to the entire matrix composed of the polymer material through a chemical bond, for example, by a polymer material having a functional group capable of reacting with the alkylene glycol residue-containing component. After constituting the matrix, this functional group can be reacted with an alkylene glycol residue-containing component. Or it can also carry out by the copolymerization etc. of the unit which comprises the matrix material as a polymeric material, and the unit which has an alkylene glycol residue containing component. Preferably, it is an embodiment performed by copolymerization.

  Such copolymerization can be realized, for example, by using a monomer constituting the matrix material and an alkylene glycol residue-containing monomer obtained by introducing an alkylene glycol residue-containing component into this monomer. The site for introducing the alkylene glycol residue-containing component into the matrix material may be either the main chain or the side chain of the polymer material, but is preferably introduced into the side chain of the polymer material. Further, the form of copolymerization at this time is not particularly limited, and may be random copolymerization or block copolymerization. In view of the homogeneity of the surface properties of the matrix, random copolymerization is preferred.

  In addition, the alkylene glycol residue-containing component may be separately introduced by bonding via a linker to the surface of the matrix for the purpose of hydrophilic modification only on the surface.

(Alkylene glycol residue-containing monomer)
When the alkylene glycol residue-containing component is introduced into the matrix by copolymerization, various compounds can be used as the alkylene glycol residue-containing monomer depending on the type of matrix material used. For example, when the matrix material is a (meth) acrylic polymer material, the compound represented by the above formula (2) can be used as the type of the polymerizable functional group that the alkylene glycol residue-containing monomer has. Here, Y in Formula (2) represents a hydrogen atom or a methyl group. Preferably Y is a methyl group.

  This solid phase material contains an alkylene glycol residue-containing component represented by formula (1), thereby adsorbing minute objects when not irradiated with light, or adsorbing substances that react with immobilized minute objects. Can be suppressed. In particular, it is useful for suppressing adsorption of minute objects when light is not irradiated on the solid surface. The adsorption suppression ability of the solid phase material is the same as that for the light immobilization amount when the light object is irradiated on the solid phase or in the vicinity thereof, except that no light is irradiated in the measurement of the light immobilization amount. The amount of light immobilization is more than 3 times the amount of adsorption when no light is irradiated when comparing the amount of adsorption of minute objects on the solid phase surface when adsorbing It is preferable. If it is 3 times or more, the immobilization site and the non-immobilization site of the minute object can be clearly distinguished, and the selective immobilization ability by light irradiation is sufficient. As a result, it is possible to increase the contrast of the minute object between the immobilized part and the non-immobilized part. More preferably, the amount of light immobilization is 4 times or more, more preferably 5 times or more than the amount of adsorption when no light is irradiated.

  The ability to suppress adsorption when light is not irradiated on a minute object is due to the alkylene glycol residue-containing component in the matrix. In the present fixing material, since the alkylene glycol residue-containing component is contained in the entire matrix, the adsorption suppressing ability when not irradiated with light can be expressed over the entire surface of the matrix. As a result, when the light of the minute object is fixed, it is possible to supply the minute object non-selectively to a region other than the fixed region. Therefore, it is suitable for immobilizing a minute object using fine patterning at the time of light irradiation.

  The solid phase material is not only excellent in the ability to suppress adsorption when not irradiated with light, but also has a sufficient ability to immobilize minute objects. That is, even though it contains a polyethylene glycol residue that imparts hydrophilicity to the solid phase material, a conventional solid phase material for light immobilization (for example, this solid phase material contains an alkylene glycol residue-containing component) It has at least almost the same immobilization ability as the same material). Therefore, the solid phase material can simultaneously realize the suppression of the adsorption of the minute object in the light non-irradiation region and the sufficient light immobilization of the minute object.

  The solid phase material is also excellent in durability against external stress. This is considered to be due to molecular mobility derived from an alkylene glycol residue-containing component. That is, the present solid phase material has higher durability and lower film thickness uniformity than the same material except that the solid phase material does not contain an alkylene glycol residue-containing component. This is particularly effective when the film is thinned.

  Furthermore, the present solid phase material can maintain the stability of a minute object, particularly a biological substance, immobilized on the surface of the solid phase. This is considered to be due to the biomolecules being prevented from being deteriorated by water molecules on the surface of the solid phase material included by the alkylene glycol residue-containing component.

  The stability retention capability for maintaining the stability of the micro object is not particularly limited, but it should be higher than that of the micro object immobilized on the same material except that this solid phase material does not contain an alkylene glycol residue-containing component. Is preferred. More preferably, the solid phase material has a stable retention capacity 1.1 times or more that of the same material except that the solid phase material does not contain an alkylene glycol residue-containing component.

  The stability of the immobilized micro object can be evaluated by, for example, photofixing the same micro object to the same material under the same conditions except that the solid phase material and the solid phase material do not contain an alkylene glycol residue-containing component. And the amount of micro-objects on the solid phase immediately after photo-immobilization or the amount of activity thereof, and the amount of micro-objects on the solid phase or the activity after a predetermined period of time after photo-fixation (for example, several days to several tens of days later) This can be done by comparing the amount. In this case, the closer the value of the minute object amount or the like immediately after the light fixation is to the minute object amount or the like after the predetermined period, the smaller the deterioration of the minute object, that is, the higher the stability of the minute object.

  “High stability of minute object” means that the immobilized state of the minute object once immobilized on the solid phase material is continued, as well as the higher-order structure inherent to the immobilized minute object, This includes that the function, physical properties, etc. are maintained or that there is little change. For example, when the minute object is an enzyme, the stability can be evaluated by detecting the enzyme activity. Alternatively, when the micro object is an antibody, the stability can be evaluated by detecting the binding ability to the antigen as its antibody activity.

(Photo-responsive component)
The photoresponsive component is a component that causes a change in molecular structure or molecular arrangement by light. The phenomenon in which molecular structure changes due to light is generally referred to as photochromism. As the photoresponsive component used in the present invention, a compound generally referred to as a photochromic compound can be used. Among them, a compound that causes photoisomerization is preferably used. In addition, there are compounds that cause changes in molecular arrangement (especially anisotropic changes) such as photo-induced orientation and photoassociation with or without molecular structure changes such as photoisomerization. As long as light immobilization on the surface of the solid phase material is possible, it can be used as the photoresponsive component of the present invention.

  As the light-responsive component, various components that have been used for such light fixation can be used. Examples of the photoresponsive component include organic compounds such as azo compounds, spiropyran compounds, spirooxazine compounds and diarylethene compounds, and inorganic materials generically called chalcogenite glass.

The photoresponsive component is preferably a compound (azo compound) having a dye structure having an azo group (—N═N—). The azo compound undergoes cis-trans isomerization by light irradiation or the like, and the movement at the molecular level by this isomerization plasticizes the matrix material to facilitate deformation. Of these, amino-type azobenzene compounds having a structure of aminoazobenzene or a derivative thereof are preferable. The amino-type azobenzene compound can be typically represented by the following formula (3).

In formula (3), R ³ and R ⁴ each independently represent a hydrogen atom or a substituent, and X represents a hydrogen atom, an electron-withdrawing substituent or an electron-donating substituent.

Examples of the substituent in R ³ and R ⁴ include alkyl group, hydroxyalkyl group, halogen atom, nitro group, amino group, carboxyl group, aryl group, allyl group, alkyl ester group, alkyl ether group, alkylamino group, alkylamide Examples include a group, an isocyanate group, and an epoxy group. In particular, when one of the substituents in R ³ and R ⁴ is an acrylic compound such as acrylic acid or (meth) acrylic acid having a polymerizable double bond at the terminal, or other double bond component, Formula (3) represents a photoresponsive polymer provided with a polymer group derived from a double bond component, as well as a double bond monomer. When one of the substituents in R ³ and R ⁴ is a polycondensation or polyaddition polymerizable component such as an isocyanate group, an amino group, a carboxyl group, or a hydroxyl group, the formula (3) represents a polymerizable monomer In addition to the above, it represents a photoresponsive monomer having a polymerizable group derived from a polymerizable component.

  Examples of the electron-withdrawing substituent in X include a cyano group, a nitro group, a sulfo group, or a sulfonic acid derivative group. Examples of the electron-donating substituent include an amino group, a dimethylamino group, An alkyl group etc. are mentioned. Among these, those having an electron-withdrawing substituent are preferable because the matrix material is considered to have a large plasticizing action by repeating cis-trans isomerization during light irradiation. In addition, the photoresponsive component which is an azo compound can be used 1 type or in combination of 2 or more types.

  Such a photoresponsive component may be added to the matrix as a component separate from the matrix material, but is present in a part (main chain or side chain) of the matrix material through a chemical bond. It is preferable. The photoresponsive component is preferably present uniformly in the matrix. By doing so, it is possible to optically immobilize the minute object at a desired site on the surface of the solid phase material.

(Micro object)
The type of the minute object is not particularly limited as long as it is tangible. For example, (1) inorganic materials such as metals, metal oxides, semiconductors, ceramics and glass, (2) organic materials such as so-called plastics, (3) biomolecular materials such as proteins, nucleic acids, saccharides and lipids, (4) One or two or more materials selected from composite materials obtained by combining two or more materials selected from the various materials (1) to (3) described above can be used. A biomolecular material is preferable.

(Biomolecule)
The biomolecule which is the subject of the present invention does not mean only one molecule, but may be an assembly of homologous molecules composed of two or more molecules or a complex with different molecules. Further, it may be an organization such as a self-organization composed of a large number of same or different molecules.

  The biomolecule is not particularly limited, and is a naturally occurring molecule that is present in an organism such as an animal, plant, microorganism, or virus, is produced by the organism, or is metabolized by the organism, or a molecule obtained by artificially modifying these. Alternatively, it may be an artificially designed molecule that does not depend on natural molecules. In addition to molecules collected from living organisms, they may be molecules produced artificially in organisms other than the organism in which the molecules originally exist, or may be artificially synthesized outside the organism, chemically synthesized, enzymes, etc. It may be a molecule synthesized by

  Typically, biomolecules include proteins, nucleic acids, saccharides, sugar chains, lipids (including lipid structures such as liposomes), biomaterials such as osteogenic materials, various biological cells and parts thereof (for example, Biological materials such as mitochondria, thylakoids, sub-mitochondrial particles, etc.), tissues and living organisms themselves (for example, yeast or Escherichia coli for alcohol fermentation or lactic acid fermentation). In addition, the biomolecules may be combined with other organic materials and / or inorganic materials when being immobilized on the solid phase. Of these, the biomolecule immobilized on the solid phase material is preferably a protein.

  As used herein, protein includes proteins, polypeptides and oligopeptides of any size, structure or function. Examples of the protein include enzymes (for example, cellulase, lipase, ATP synthase, enzymes involved in alcoholic fermentation or lactic acid fermentation, etc.), antigens, antibodies, membrane proteins, proteoglycans, lectins, or cell membrane receptors. Antibody also refers to a naturally occurring or wholly or partially synthetically produced immunoglobulin. Also included are all derivatives thereof that retain specific binding ability. The nucleic acid may be single-stranded or double-stranded. It may contain DNA, RNA, DNA / RNA hybrids, DNA-RNA chimeras, bases and other modifications, whether artificial or natural. Further, the chain length is not particularly limited. Further, fine particles (for example, polymer fine particles, magnetic particles, silica particles, etc.) bound or adsorbed with biomolecules such as these proteins may also be included.

  Note that photoresponsive components and minute objects that can be used in the present invention are disclosed in JP 2003-329682 A, JP 2004-93996 A, JP 2004-251801 A, and JP 2007-51998 A. The described carriers and light immobilization materials can be used. In addition, this specification includes Japanese Patent Laid-Open Nos. 2003-329682, 2004-93996, 2004-251801, 2006-233004, 2006-321719, and 2007-. All matters described in the 51998 public information shall be incorporated by reference.

(Use)
The solid phase material of the present invention can be used for functional analysis of minute objects such as expression, interaction, post-translational modification of target minute objects, analysis and function analysis of other substances using minute objects, reaction, adsorption, etc. Can be used. That is, the solid phase material of the present invention can be used for diagnosis and treatment as a so-called protein chip, DNA chip, antibody chip and the like. In addition, various reactors such as bioreactors, microbioreactors, ATP synthesis bioreactors that perform enzyme reactions, etc., removal of predetermined substances such as reaction catalysts and deodorants, catalysts for reducing VOC (volatile organic compounds), observation It can also be used as a substrate for immobilizing an object (for example, a sample for AFM (atomic force microscope)). In particular, the solid phase material is suitable for use as a microreactor because it can immobilize a minute object using fine patterning during light irradiation.

(Solid phase)
The solid phase body of the present invention is obtained by holding the micro object on the above-described solid phase material for light immobilization by light immobilization. As the solid phase material in this solid phase body, the solid phase material of the present invention in the various forms described above can be applied as it is. Therefore, according to the present solid phase body, it is possible to obtain a solid phase body in which a micro object is immobilized on the surface of the solid phase material, thereby obtaining the various advantages described above in the solid phase material of the present invention. it can. That is, this solid phase body is excellent not only in detection accuracy for interaction with or reaction with a minute object, but also in structural and mechanical characteristics. In addition, the stability of the micro object once immobilized on the solid phase material can be maintained.

  Such a solid phase of the present invention is used as a so-called protein chip, DNA chip, microreactor, etc. for analysis and analysis utilizing the function of a minute object immobilized on the surface of a solid phase material, or for diagnosis or treatment. be able to.

(Method for producing solid phase)
The method for producing a solid phase body of the present invention includes a preparation step for preparing the solid phase material of the present invention described above, and irradiating the micro object existing on or near the surface of the solid phase material with light. Holding on the solid surface. In addition, the structure, component, shape, application, etc. of the already described solid phase material of the present invention can be applied as they are to the solid phase material in the present production method. Therefore, the preparation process in this manufacturing method can include various manufacturing processes for manufacturing the above-mentioned various forms of the solid phase material of the present invention.

(Holding process)
In the holding step of the manufacturing method, first, a micro object is supplied to the prepared solid phase material. A method for supplying the micro object to the surface of the solid phase material is not particularly limited, but it is preferable to supply the micro object through the liquid medium in a state of being dissolved or suspended in the liquid medium. Moreover, when supplying a micro object, you may supply selectively to the surface of a solid-phase material, and you may supply non-selectively. Here, the selective supply includes, for example, an aspect in which a large number of minute objects are arranged according to a specific distribution pattern. As an example of such patterning, there is an aspect in which spots of minute objects are supplied in an array. On the other hand, the non-selective supply includes supplying to an unspecified region on the surface of the solid phase, for example, a region including a non-immobilized portion of a micro object (for example, the entire surface of the solid phase).

  The minute object supplied to the solid surface can be fixed (light-fixed) to the solid surface by being irradiated with light. Here, light immobilization means immobilizing a micro object supplied to the surface of a solid phase material containing a photoresponsive component by causing photoisomerization or photodeformation of the photoresponsive component by light irradiation. Say.

  In light fixation, since fixation is possible even by irradiation with visible light having low energy, it is possible to realize fixation that suppresses or avoids adverse effects on a biomolecule as a minute object. Therefore, unlike the covalent bond immobilization method, the biomolecule can be immobilized while maintaining the three-dimensional structure and good activity.

  The light irradiation method for light fixation is not particularly limited. Irradiation may be performed so that arbitrary light such as various kinds of propagating light, near-field light, or evanescent light reaches or near the solid-phase surface where the minute object exists. Moreover, although light irradiation may be performed with respect to the whole surface of a solid-phase, it can also be selectively performed with respect to the predetermined area | region which should be fix | immobilized on a solid-phase surface. In particular, since this solid-phase material suppresses the adsorption of minute objects when light is not irradiated over the entire surface of the solid phase, the solid-state material reliably fixes the irradiation light in the irradiation region, while the light non-irradiation region. It is possible to sufficiently suppress the adsorption of minute objects on the surface.

  In order to selectively perform the light irradiation, for example, by giving a certain distribution to the irradiation region of the irradiation light with respect to the solid phase surface, the light can be immobilized according to a specific distribution pattern. For example, a mode using scanning with a photomask, interference light, or laser light can be mentioned. By such a method, the immobilization region of the minute object can be immobilized, for example, so as to be a flow path in the microreactor.

  After the micro object is optically fixed on the solid surface, a step for washing the solid surface may be performed. In this solid phase body, the micro object is securely fixed to the surface of the solid phase despite the fact that the solid phase material contains an alkylene glycol residue. There is little outflow.

  In addition, as light irradiation for light fixation, the irradiation light and light as described in Unexamined-Japanese-Patent No. 2003-329682, Unexamined-Japanese-Patent No. 2004-93996, Unexamined-Japanese-Patent No. 2004-251801, and Unexamined-Japanese-Patent No. 2007-51998 are mentioned. An irradiation method can be employed. Regarding light immobilization, JP-A No. 2003-329682, JP-A No. 2004-93996, JP-A No. 2004-251801, JP-A No. 2006-233004, JP-A No. 2006-321719 and JP-A No. 2007- No. 51998 has already been disclosed by the present applicant, and these methods can also be applied to light immobilization in the present invention.

(How to use solid phase)
The method of using the solid phase of the present invention comprises a step of causing an interaction between a micro object supplied with a test sample to a micro object fixed to the solid phase and a component in the test sample, and the interaction Detecting. For the solid phase material in this method of use, the configurations, components, shapes, uses, etc. of the solid phase material and solid phase body of the present invention already described can be applied as they are. Therefore, according to this method of use, since the solid phase body in which the adsorption of the minute object in the region other than the immobilization region of the minute object is used is used, the interaction with the minute object can be detected with high accuracy. .

  Hereinafter, the present invention will be described with specific examples, but the present invention is not limited to the specific examples illustrated below.

(1) Synthesis of solid phase material for light immobilization The compound shown in “Chemical Formula 4”, methyl methacrylate (manufactured by Wako Pure Chemical Industries), and polyethylene glycol methacrylate (manufactured by Aldrich) are radically copolymerized at various ratios, The polymer material shown in “Chemical Formula 5” was synthesized. Here, ones having different lengths of polyethylene glycol side chains (value of k in Chemical Formula 5, where k represents an average value) and copolymerization ratios (values of n, m, and l in Chemical Formula 5) were synthesized ( Test examples 1 to 3). The polyethylene glycol ester of methacrylic acid used is a mixture of different lengths.

Details of the copolymerization reaction are as follows. The monomer mixture (total 10 mmol) 2,2′-azobisisobutyronitrile (AIBN, 40 mg, 0.24 mmol) of each test example and comparative example 1 shown in Table 1 was dissolved in DMF (10 ml). Nitrogen bubbling was performed at room temperature for 30 minutes, and then the polymerization reaction was performed by stirring at 60 to 70 ° C. for 4 hours under a nitrogen stream. After the reaction solution was cooled to room temperature, the reaction solution was slowly added dropwise to methanol (500 ml). The precipitated polymer was collected by filtration and dissolved in acetone (10 ml). The acetone solution was slowly added dropwise to methanol (500 ml) to precipitate the polymer again. The polymer was recovered by filtration and vacuum dried at 40 ° C. overnight to obtain a solid phase material for photo-immobilization. The copolymerization ratio of the obtained solid phase material was determined by measuring ¹ H-NMR using a superconducting Fourier transform nuclear magnetic resonance apparatus (JNM-LA500) manufactured by JEOL in a deuterated chloroform solvent. Calculated. The molecular weight was measured with a chloroform solvent using GPC (Shodex GPC-101, column: K-800RL + K-805L × 2) manufactured by Showa Denko KK, and the molecular weight in terms of polystyrene was calculated.

Table 1 shows the copolymerization ratio, average molecular weight, and polyethylene glycol residue content (% by mass) of the solid phase material for photofixation. Note that the content of polyethylene glycol residues, means the content of the alkylene glycol residue represented by among _{-O- (CH 2 -CH 2 -O)} k -CH 3 of Compound 5. Comparative Example 1 is a solid phase material for light immobilization without introduction of polyethylene glycol.

  50 mg of each of the solid phase materials of Test Examples 1 to 3 and Comparative Example 1 was dissolved in 4 ml of pyridine, and about 1 ml of each of these solutions was dropped on a slide glass substrate. And the solvent was removed by rotating a slide glass substrate at 4000 rpm, and the polymer film was produced. All of these polymer films had a film thickness of about 40 nm.

(2) Evaluation of antibody immobilization and adsorption In this example, the degree of antibody adsorption was evaluated as follows. That is, Cy5-labeled antibodies (Cy5-IgG: Anti-IgG (Fc), Mouse, Goat-Poly, Cy5 [CHEMICON: AP127S]) were allowed to stand (adsorb) in the dark without being irradiated with light. The amount of antibody on the solid phase material was compared with the case.

The photofixation of the antibody was performed as follows. Various concentrations (0, 10, 20, 50, 100, 200, 300, 400 and 500 ng / ml) of Cy5-IgG in TPBS (0.01% Tween20 PBS, pH = 7.4) solution were placed on a slide glass substrate. 1 μl each was spotted on the polymer film prepared in (1). After drying the spot in vacuum, it was irradiated with light for 30 minutes using a blue LED (20 mW / cm ² ). Thereafter, the slide glass was washed 3 times for 5 minutes using TPBS.

  For antibody adsorption, Cy5-IgG in TPBS solution was spotted and dried in the same manner as in the above light fixation, and then left in the dark for 30 minutes without light irradiation. Thereafter, the slide glass was washed 3 times for 5 minutes using TPBS.

  Fluorescence intensity was measured for various glass slides after washing. The measurement was performed using a confocal laser microscope (Affymetrix: 428 Array Scanner). The fluorescence intensity at the Cy5 detection wavelength was measured at an excitation wavelength of 633 nm and a detection wavelength of 660 to 680 nm.

Table 2 shows the amount of adsorption in each test example when the amount of adsorption in Comparative Example 1 (the amount of antibody at the time of non-irradiation) is 100, and the ratio of the light fixation amount (the amount of antibody at the time of irradiation) to the amount of adsorption. About adsorption amount, as shown in Table 2, it was 1/2 or less of the comparative example 1 in any of Test Examples 1-3 which introduce | transduced polyethyleneglycol. In particular, in Test Examples 1 and 3, the adsorption amount was 1/5 or less that of the Comparative Example. From these facts, it was found that the amount of adsorption at the time of non-irradiation can be reduced by introducing polyethylene glycol. In addition, it is considered that the background signal when using a solid phase on which a minute object is immobilized can be reduced.

  Moreover, when the ratio of the light immobilization amount with respect to the adsorption amount at the time of non-light irradiation was seen, in each of Test Examples 1 to 3, they were larger than Comparative Example 1 and were 5, 3, and 4, respectively. Particularly, in Test Example 1, the value was 2.5 times that of Comparative Example 1, and in Test Example 3, the value was twice that of Comparative Example 1. From these facts, it was found that the ability of selective immobilization by light irradiation can be enhanced by introducing polyethylene glycol. As a result, it is possible to increase the contrast of the minute object between the light irradiation part and the light non-irradiation part. Therefore, the solid phase material into which polyethylene glycol is introduced is suitable for use in patterning a micro object using selective light irradiation.

(3) Evaluation of durability of thin film (durability against external stress) In order to evaluate the durability of the thin film by introduction of polyethylene glycol, the thin films of Test Example 1 and Comparative Example 1 were mixed with TPBS (0.01% Tween 20 PBS, pH = 7.4) After being immersed in the solution for 72 hours, a cover glass was placed on the surface of the thin film, and the cover glass was shifted several times in the front-rear direction. The state of the subsequent film was observed with a dark field optical microscope. The result is shown in FIG. The white spots in the photomicrographs are the parts that scatter the light, and are where alterations such as cracking of the film are occurring. Prior to treatment, almost no white spots were observed on either film. However, in Comparative Example 1 in which polyethylene glycol was not introduced, white spots were observed in the entire visual field of the processed thin film. On the other hand, in Test Example 1 in which polyethylene glycol was introduced, there were few white spots. From this, it was found that the durability against external stress of the film was improved by introducing polyethylene glycol.

(4) Evaluation of protein stabilization In order to evaluate the stability of the biomolecule immobilized on the thin film into which polyethylene glycol was introduced, the antibody was immobilized on the thin film of Test Example 1 and Comparative Example 1, and 15 ° C. at 4 ° C. Antibody activity after storage in the dark for days was evaluated.

  The stability was evaluated by immobilizing an anti-goat-rabbit antibody (anti-goat-IgG: Anti-IgG (H + L), Goat, Rabbit-Poly [Bethl Laboratories: A50-100A]), and then anti-goat-IgG. Anti-antibody reaction was performed using Cy5-labeled antibodies (Cy5-IgG: Anti-IgG (Fe), Mouse, Goat-Poly, Cy5 [CHEMICON: AP127S]), and Cy5 fluorescence intensity was measured. At this time, with respect to Test Example 1 and Comparative Example 1, the fluorescence intensity (A) when the reaction with Cy5-IgG was performed immediately after the anti-goat-IgG was immobilized, and the anti-goat-IgG was immobilized. The fluorescence intensity (B) in the case of reaction with Cy5-IgG after storage in the dark at 15 ° C. for 15 days was compared. In addition, it is shown that the closer the values of A and B are, the less the antibody is deteriorated by storage at 4 ° C., that is, the higher the stability of the antibody.

The photofixation of the antibody was performed as follows. Various concentrations (0, 10, 20, 50, 100, 200, 300, 400 and 500 ng / ml) of anti-goat-IgG in TPBS (0.01% Tween20 PBS, pH = 7.4) 1 μl each was spotted on the polymer film produced on the substrate. After drying the spot in vacuum, it was irradiated with light for 30 minutes using a blue LED (20 mW / cm ² ). Thereafter, the slide glass was washed 3 times for 5 minutes using TPBS.

  The antigen-antibody reaction was performed as follows. 40 μl of 1% gelatin TPBS containing 1 μg / ml of Cy5-IgG was dropped onto the center of the slide glass, and the reaction was carried out by covering the gap cover glass. The reaction temperature was 25 ° C. and the reaction time was 30 minutes. After the reaction, the slide glass was washed twice for 1 minute with TPBS.

  The fluorescence intensity was measured by using a confocal laser microscope (Affymetrix: 428 Array Scanner). The fluorescence intensity at the Cy5 detection wavelength was measured at an excitation wavelength of 633 nm and a detection wavelength of 660 to 680 nm.

In each of Test Example 1 and Comparative Example 1, the value of the fluorescence intensity B when the fluorescence intensity A was 100 was measured. The results are shown in Table 3. As shown in Table 3, in Comparative Example 1, the fluorescence intensity B was 70, whereas in Test Example 1, the fluorescence intensity B was 80. From this, it was found that by introducing polyethylene glycol, the ability of the antibody to bind to the antigen is kept higher, and the stability of the immobilized antibody is improved.

Explanatory drawing showing the evaluation result of durability of this solid-phase material. 1A shows Test Example 1 and FIG. 1B shows Comparative Example 1. FIG.

Claims (19)

A solid phase material for light immobilization that immobilizes a minute object by light irradiation,
A solid phase material comprising a matrix of the solid phase material containing a photoresponsive component that causes a change in molecular structure or a molecular arrangement by light and an alkylene glycol residue-containing component represented by the following formula (1).
(However, R ¹ represents a hydrogen atom, a substituent, or a linking group connected to the constituent components of the matrix, and R ² represents a hydrogen atom or an alkyl group having 1 to 10 carbon atoms. Represents an alkylene group having 1 to 10 carbon atoms, and n represents an integer of 1 to 100.)   The solid phase material according to claim 1, wherein the micro solid object is fixed to the solid phase material by deforming the solid phase material for light immobilization by light irradiation.   The solid phase material according to claim 1 or 2, wherein the amount of light immobilized by light irradiation is at least 3 times the amount of adsorption when light is not irradiated.   The solid phase material according to any one of claims 1 to 3, wherein durability against external stress is improved as compared to the same solid phase material except that the alkylene glycol residue-containing component is not contained.   The solid phase according to any one of claims 1 to 4, wherein the minute object fixed to the solid phase material has higher stability than the same solid phase material except that the alkylene glycol residue-containing component is not contained. material.   The solid phase material according to any one of claims 1 to 5, wherein Z represents an alkylene group having 1 to 4 carbon atoms.   The solid phase material according to claim 1, wherein n is an integer of 4 or more and 25 or less. The matrix is a polymer;
Content of the alkylene glycol residue represented by -O- (ZO) n-R < ² > in the said alkylene glycol residue containing component with respect to the mass of the said matrix is 50 mass% or less. The solid phase material according to any one of the above.   The solid phase material according to claim 8, wherein the content of the alkylene glycol residue is 30% by mass or less.   The solid phase material according to claim 1, wherein the matrix is obtained by polymerizing a monomer composition containing a photoresponsive component-containing monomer and an alkylene glycol residue-containing monomer. The said polyalkylene glycol residue containing monomer is a solid-phase material of Claim 10 represented by the following formula | equation (2).
(However, Y represents a hydrogen atom or a methyl group.) Z represents an alkylene group having 1 to 4 carbon atoms;
The matrix is a polymer;
^2. The content of an alkylene glycol residue represented by —O— (Z—O) n—R ² in the alkylene glycol residue-containing component with respect to the mass of the matrix is 30% by mass or less. Solid phase material.   The solid material according to claim 1, wherein the micro object is a biomolecule.   The solid phase material according to claim 1, wherein the micro object is a protein.   The solid phase material according to any one of claims 1 to 14, which is used for immobilization by selective light irradiation. A solid body on which a minute object is immobilized,
The said micro object is a solid-phase body currently hold | maintained on the surface of the solid-phase material in any one of Claims 1-15. A method for producing a solid body in which a minute object is immobilized,
A preparation step of preparing the solid phase material according to any one of claims 1 to 15,
Holding the micro object on the solid phase by irradiating the micro object existing on or near the surface of the solid phase material; and
A manufacturing method comprising:   The manufacturing method according to claim 17, wherein the holding step supplies the micro object non-selectively to the surface of the solid phase material and selectively irradiates a predetermined region of the surface of the solid phase material with light. . A method of using a solid phase in which a minute object is immobilized,
Supplying a test sample to the micro object fixed to the solid phase body according to claim 16 to cause an interaction between the micro object and a component in the test sample;
Detecting the interaction;
Use method comprising.