JP2010260032A - Adsorptive carrier which has region of selective adsorption of target molecule, and production method of the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a target-molecule-adsorptive carrier which can realize good selectivity and good adsorptivity. <P>SOLUTION: A target-molecule-adsorptive carrier having one or two or more regions of selective adsorption of one or two or more kinds of molecules is produced by feeding the target molecules into the surface of a solid-phase material to produce an interaction with the target molecules on the surface to allow them to be adsorbed by the surface and removing the target molecules from the surface of the solid-phase material. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a solid phase material having a surface having selective adsorption ability and a method for producing the same.

  In recent years, molecular recognition has attracted attention as one of the functions of polymer materials. Molecules that are desired to be target molecules vary from low-molecular compounds to proteins, and their application to sensors and column fillers is expected by selectively recognizing target molecules. In order to impart molecular recognition ability, it is usually necessary to form in the solid phase material a template that records the shape of the molecule to be the target molecule and the position of the functional group. In order to obtain such a molecule recognition material, a general method is to prepare a solid phase material into which a target molecule has been introduced in advance, and subsequently remove the target molecule and go through a process.

  One method for synthesizing such molecular recognition materials is to mix and interact with a monomer having a functional group that interacts with the target molecule, then cross-link the polymerizable group of the monomer, and then remove the target molecule. Thus, a method (molecular imprinting method) for forming a void (template) having a predetermined shape of a target molecule and a position where it interacts with the target molecule is known. For example, when a low molecular compound is used as a target molecule, it is known to polymerize a polymerizable monomer or a crosslinkable monomer in the presence of the low molecular compound (Patent Document 1). In addition, when a protein or the like is used as a target molecule, an alkoxysilane compound having a functional group that interacts with the protein is formed on the surface of the carrier or in the vicinity after the protein is held on a suitable carrier surface by a covalent bond. Methods for polymerizing and subsequently removing proteins are known (Non-Patent Documents 1 and 2).

  As another synthesis method, a method of preparing a material that recognizes a target molecule by mixing a polymer dissolved in a solvent and a target molecule to prepare a dispersion and then depositing the polymer has been reported (Patent Document). 2). The target molecule in this technique is a low-molecular functional compound, and it is possible to prepare a molecular recognition material of any shape by surrounding the functional compound with a polymer and then precipitating the polymer.

Japanese National Patent Publication No. 8-506320 JP 2008-101052 A

ShuSheng Zhang et al., `` Molecular imprinted polymer grafted on polysaccharide microsphere surface by the sol-gel process for protein recognition '', Talanta, 2008, 74, 1247-1255. Kengo Sakaguchi et al., `` A method for the molecular imprinting of hemoglobin on silica surfaces using silanes '', Biomaterials, 2005, 26, 5654-5571.

  In the conventional molecular imprinting method, in the presence of a target molecule, the constituent monomers of the molecular recognition material are polymerized in situ to synthesize a molecular recognition material having a target molecule template. . In order to increase the recognition ability of the target molecule, it is necessary to use a monomer that interacts strongly with the target molecule, to increase the crosslinking density, and to transfer the shape and surface properties of the target molecule. However, increasing the crosslink density makes it difficult to remove the target molecules and to enter the target molecules into the material. On the other hand, by weakening the interaction and reducing the crosslink density, it is easy to remove the target molecule and enter the target molecule into the solid phase material, but the selectivity of the target molecule is reduced. As described above, it is impossible to achieve both high molecular recognition ability and target molecule removal by the molecular imprinting method, and it is difficult to substantially realize a good adsorption rate by molecular recognition.

  In general, the shape of biomolecules such as proteins changes due to external factors. Moreover, the environment in which the original biological activity is exhibited is limited to a certain range. Therefore, when such a biomolecule is used as a target molecule, the conditions for using the molecular recognition material, that is, the conditions for trying to adsorb the target molecules are significantly different from the conditions for forming the target molecule template. Then, there exists a possibility that an adsorption rate may fall. In addition, when the conditions at the time of template formation and the environmental conditions for exerting the biological activity of the target molecule are greatly different, there is a possibility that the active target molecule cannot be adsorbed. As a result, the intended target molecule adsorption rate cannot be realized.

  As described above, the target molecule recording process by polymerization or precipitation in the conventional method for synthesizing molecular recognition materials prioritizes the condition of forming a template, and tries to adsorb actual target molecules. Conditions are not considered at all.

  Furthermore, the molecular recognition material according to the conventional method is merely provided with a molecular recognition region for a single target molecule at random in the molecular recognition material, and a plurality of molecular recognition regions for the same or different target molecules are included in the solid-state material. It was not possible to form the surface in a controlled state.

  An object of the present invention is to provide a carrier for adsorbing target molecules capable of realizing good selectivity and adsorption rate. Another object of the present invention is to provide an adsorption carrier provided with a selective adsorption region for target molecules in a position-controlled state.

  As a result of intensive studies to achieve the above object, the inventors of the present invention have obtained a solid phase obtained by removing a target molecule after interacting and adsorbing a target molecule with a previously prepared solid support surface. The carrier surface has been found to have the property of selectively re-adsorbing target molecules, and the present invention has been completed. According to the present invention, the following means are provided.

  According to the present invention, the following step (a) supplying a target molecule to the surface of a solid phase material to cause interaction with the target molecule on the surface and adsorbing to the surface; (b) And a step of removing the target molecule from the surface of the solid phase material. A method for producing a carrier for adsorption of a target molecule comprising one or more selective adsorption regions of one or more kinds of target molecules is provided. Is done.

  The selective adsorption region may have at least a part of the deformed and / or changed properties of the solid phase material obtained by the adsorption of the target molecule, and is complementary to a part of the target molecule. It is good also as what has a space | gap.

  Further, the step (a) can be a step including immobilizing the target molecule on the surface of the solid phase material containing a photoresponsive material by light irradiation. The photoresponsive material may contain a polymer material, or may contain azobenzene or an azobenzene derivative as a photoresponsive component. Further, the azobenzene derivative may be push-pull azobenzene or aminoazobenzene.

  In the step (b), the removal medium containing any one selected from water and an aqueous solution containing one or more selected from surfactant, salt, acid and alkali is used as the solid phase material. It is good also as a process of supplying to the surface and removing the target molecule.

  Furthermore, the average size of the target molecule may be 1 nm or more and 100 μm or less, and the target molecule may be a biomaterial or a biomaterial.

  In the production method of the present invention, the step (a) causes the interaction with one or more kinds of the target molecules in one or more selected regions of the surface of the solid phase material. It can also be set as the process of adsorb | sucking a target molecule.

  According to the present invention, there is also provided a solid carrier for adsorption of target molecules obtained by any one of the production methods described above. In addition, according to the present invention, a solid phase material containing a photoresponsive material and one or more target molecules on the surface of the solid phase material are photofixed and then the target molecules are removed. There is provided a carrier for adsorption of a target molecule, comprising one or more selective adsorption regions. In the adsorption carrier of the present invention, two or more selective adsorption regions may be arranged on the surface of the solid phase material.

  In addition, according to the present invention, there are also provided a detection device, a cell growth device, a protein recovery device, and a microreactor including a target molecule adsorption carrier. Furthermore, according to this invention, the material for adsorption carriers of a target molecule provided with the solid-phase material containing a photoresponsive material is also provided.

It is a figure which shows an example of the manufacturing process of the support | carrier for adsorption | suction of this invention. It is a figure which shows the manufacture and evaluation flow of the support | carrier for adsorption | suction in an Example. It is a figure which shows the kind and size of protein which were used in the Example. The numbers in the table correspond to the spot numbers in the spot layout shown in FIG. In Example 1, BSA was used, and in Examples 2 and 3, HSA was used. It is a figure which shows the layout of the protein spot in an Example. Spot number 3 represents BSA in Example 1 and HSA in Examples 2 and 3. It is a figure which shows the result of having evaluated re-adsorption of IgG with fluorescence intensity. Spot 3 is BSA. It is a figure which shows the result of having evaluated re-adsorption of IgG with fluorescence intensity. Spot 3 is HSA. It is a figure which shows the result of having evaluated the re-adsorption of HSA by the fluorescence intensity. Spot 3 is HSA. It is a figure which shows the result of having evaluated re-adsorption of IgG with fluorescence intensity. Spot 3 is HSA. It is a figure which shows the result of having evaluated the re-adsorption of HSA by the fluorescence intensity. Spot 3 is HSA. It is a figure which shows the result of having evaluated re-adsorption of IgG with fluorescence intensity. Spot 3 is HSA. It is a figure which shows the result of having evaluated the re-adsorption of HSA by the fluorescence intensity. Spot 3 is HSA.

  In the present invention, after target molecules interact with and adsorb on the surface of the solid phase material, the target molecules are removed from the surface of the solid phase material, and the target molecules are selectively recognized and adsorbed on the surface. A solid phase material having a region is provided, and this is further provided as an adsorption carrier for target molecules. The present inventors have found that the target molecule can be adsorbed on the surface of a solid phase material prepared in advance, so that a characteristic that allows the target molecule to be re-adsorbed on the surface of the solid phase material can be recorded. . That is, the present inventors adsorb the target molecules on the surface of the solid phase material prepared in advance, and then remove the target molecules from the surface of the solid phase material, the target molecule adsorption region after the target molecule removal, It was found that the target molecule can be used as a selective adsorption region of the target molecule that can re-adsorb.

  In the present specification, the selective adsorption region is a region having at least one site that selectively recognizes and adsorbs one target molecule. The selective adsorption region is a region specific to one target molecule.

  According to the present invention, since the target molecule is held with respect to the solid phase material prepared in advance, the step of forming the solid phase material simultaneously with the formation of the target molecule template in the presence of the target molecule is not required. That is, there is no need for a monomer polymerization step in the presence of the target molecule or a step of solidifying the polymer in a state where the target molecule is surrounded by the polymer solution. For this reason, the improvement of the molecular recognition ability and the ease of removal of the target molecule, which have been problems in the past, can both be achieved. In addition, since the solid phase material is prepared in advance, the adsorption characteristics of the target molecule can be recorded on the solid phase material and the target molecule can be removed under more relaxed conditions. For this reason, in an environment in which the function of the target molecule can be exhibited, the target molecule can be removed by interacting with and adsorbing on the surface of the solid phase material without impairing the molecular recognition ability recorded on the solid phase material. Therefore, according to the present invention, it is possible to provide an adsorption carrier that can adsorb target molecules with higher selectivity and adsorption rate. Further, it is possible to provide an adsorption carrier that can recognize and adsorb target molecules under desired conditions.

  Further, according to the present invention, since a solid phase material prepared in advance is used, target molecules are supplied in a selected region of the surface, or an interaction with the surface of the solid phase material is caused only in the selected region. As a result, a selective adsorption region can be formed at an arbitrary position on the surface of the solid phase material. For this reason, a plurality of selective adsorption regions that adsorb the same or different target molecules can be easily formed in an arbitrary region on the surface of a single solid phase material. A plurality of selective adsorption regions can also be arranged.

  Furthermore, according to the present invention, the adsorption step of the present invention is an aspect in which target molecules are supplied to the surface of a solid phase material containing a photoresponsive material or in the vicinity thereof and irradiated with light to be adsorbed on the surface of the solid phase material. be able to. According to such an adsorption process, physical deformation and surface property changes due to adsorption of target molecules can be easily caused on the surface of the solid phase material, and such changes can be easily performed after removal of the target molecules. Can be maintained. Such recording performance of target molecule adsorption characteristics makes it possible to provide a solid support having high recognition ability. Furthermore, by light irradiation, a plurality of selective adsorption regions can be easily formed at an arbitrary position on the surface of the solid phase material.

  Hereinafter, various embodiments of the present invention will be described in detail with reference to the drawings as appropriate. FIG. 1 is a diagram illustrating an outline of an embodiment of the present invention, and is a diagram illustrating an example of an adsorption process and a removal process by light irradiation.

(Method for producing adsorption carrier)
The method for producing an adsorption carrier of the present invention can produce an adsorption carrier that selectively adsorbs target molecules. That is, an adsorption carrier including a solid phase material having a selective adsorption region for target molecules can be produced. The production method of the present invention includes a step of supplying target molecules to the surface of a solid phase material, causing interaction with the target molecules to occur on the surface, and adsorbing to the surface (hereinafter simply referred to as an adsorption step). A step of removing the target molecule from the surface of the solid phase material (hereinafter simply referred to as a removal step).

(Adsorption process)
The adsorption step can be a step of supplying the target molecule to the surface of the solid phase material and causing interaction with the target molecule on the surface to adsorb the target molecule on the surface. FIG. 1A shows a process of adsorbing (photofixing) a target molecule on the surface of a solid phase material by light irradiation.

(Solid phase material)
The solid phase material used in the adsorption step is prepared prior to the target molecules being adsorbed. A solid phase material is a material that has the ability to adsorb and cause a complementary interaction with a target molecule on its surface under certain conditions. The solid phase material is preferably a material having such a property that the target molecule can be adsorbed on the surface only in a portion in contact with the target molecule under the certain condition in which the complementary interaction occurs. That is, it is preferable that a specific complementary interaction is generated at the contact portion of the target molecule with the solid phase material.

  Here, the complementary interaction is preferably accompanied by the following change in the solid phase material, for example. One change is a deformation that forms a void complementary to a part of the target molecule on the surface of the solid phase material. According to the interaction accompanied by such a change, a selective adsorption region (void) corresponding to the shape of the target molecule can be formed on the surface of the solid phase material. Another change is a property change that improves the affinity to the target molecule on the surface of the solid phase material. According to the interaction accompanied by such a change, a selective adsorption region corresponding to the functional group, polarity, charge or configuration of these on the surface of the target molecule can be formed on the surface. For example, such property change may be one or two selected from hydrophilic interaction, hydrophobic interaction, electrostatic interaction, dipole interaction and van der Waals force between the solid phase material and the target molecule. Achieved by more than one action. For example, the hydrophilic interaction is derived from a hydroxyl group, amino group, amide group, carboxyl group, sulfonic acid group, phosphoric acid group, etc. in the solid phase material, and the hydrophobic interaction is an alkyl group in the solid phase material, It can be derived from a hydrocarbon group such as an aryl group.

  The complementary interaction preferably does not involve the formation of a covalent bond between the surface of the solid phase material and the target molecule, and the formation of a covalent bond within the solid phase material and within the target molecule. It is preferable not to involve. This is because if a covalent bond is generated in the target molecule, the solid phase material, or the like due to the interaction, the structure and function of the target molecule may change during the bond formation and the target molecule removal, which is not preferable. Preferably, at least some of these complementary interactions are those that remain after the target molecule is removed from the solid phase material.

(Photoresponsive material)
The solid phase material preferably includes a photoresponsive material, although it depends on the target molecule adsorption method. By including the photoresponsive material, the target molecule can be easily adsorbed on the surface of the solid phase material by using light irradiation in the adsorption process, as will be described later. It is considered that the light irradiation can cause a highly complementary interaction between the target molecule and the surface of the solid phase material.

  As the photoresponsive material, a material containing a photoresponsive component can be used. The photoresponsive material may have a photoresponsive component in a suitable matrix. The material of the matrix (matrix) is not particularly limited as long as the matrix can hold the photoresponsive component so that it can be photofixed. 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 holding ability of the photoresponsive component in the matrix, the matrix is preferably a resin or a resin composition.

  The photoresponsive component is a component that causes a change in molecular structure or molecular arrangement by light. The phenomenon in which the molecular structure is changed by 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 known for light fixation can be used. For example, there are photoisomerization compounds such as components that cause trans-cis photoisomerization, for example, organic compounds such as azo compounds, spiropyran compounds, spirooxazine compounds, diarylethene compounds, and inorganic materials collectively called chalcogenite glass Etc.

  Among these, as the photoresponsive component, it is preferable to use an azo compound having a dye structure having an azo group (—N═N—). This is because 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 photoresponsive layer 4 to facilitate deformation. Examples of the azo compound include one or more selected from azobenzene or azobenzene derivatives. Examples of the azobenzene derivative include aminoazobenzene and amino-type azobenzene compounds having a derivative structure. The amino-type azobenzene compound can typically be represented by the formula (1). In formula (1), X represents a hydrogen atom or various substituents such as an electron-withdrawing substituent, and preferably represents a hydrogen atom.

As the azobenzene derivative, for example, as shown in Formula (2) and Formula (3), an electron-withdrawing substituent (—CN, —NO ₂ ) is present on one benzene ring of the azobenzene skeleton, and electron donation is performed on the other. Compound (push-pull type azobenzene) to which an ionic substituent (—NR ¹ R ² ) is bonded. Such a compound is preferable because it is considered to have a large plasticizing action (deformation action) by repeating cis-trans isomerization during light irradiation.

  In formulas (1) to (3), R1 and R2 each independently represent a hydrogen atom or a substituent. In the formulas (1) to (3), the substituent is an alkyl group, hydroxyl group, hydroxyalkyl group, halogen atom, nitro group, amino group, carboxyl group, aryl group, allyl group, alkyl ester group, alkyl ether group, alkyl group. An amino group, an alkylamide group, an isocyanate group, an epoxy group, etc. can be illustrated. In particular, when one of the substituents in R1 and R2 is an acrylic compound such as acrylic acid or methacrylic acid having a polymerizable double bond or the like at the terminal and other double bond components, the formula (1) -(3) represents a photopolymer having a polymerizable group derived from the double bond component, as well as a double bond monomer. When one of the substituents in R1 and R2 is a polycondensation or polyaddition polymerizable component such as an isocyanate group, an amino group, a carboxyl group, or a hydroxyl group, the formulas (1) to (3) are In addition to representing a polymerizable monomer, it represents a photoresponsive monomer having a polymerizable group derived from the polymerizable component. The photoresponsive component which is an azo compound can be used alone or in combination of two or more.

  In addition, the photoresponsive component may be connected to a constituent material of the matrix such as a resin through a covalent bond or the like and may be a part thereof, or may be a separate component dispersed in the matrix. From the viewpoint of ensuring processability and plasticity of the photoresponsive layer, the solid phase material may be a polymer material containing a photoresponsive component in a monomer unit or a composition containing the polymer material. preferable. When the photoresponsive material including the photoresponsive component includes a polymer material, in the formula (1) and the like, the photoresponsive component is, as in the case where R1 or R2 is a linking group to a polymerization group. It is preferable that the polymer material is present in a part (main chain, side chain or modifying group) of the polymer material.

  As the matrix, various thermoplastic or thermosetting polymers can be used without any particular limitation. Such a polymer material is preferably used in consideration of introduction by copolymerization of a photoresponsive component or the like. Therefore, for example, (1) an acrylic polymer, a methacrylic monomer, a vinyl polymer, a styrene polymer, which is a polymer of a polymer containing a double bond such as an ethylenic monomer, and (2) the weight of a polymerizable functional group Examples thereof include addition-polymerizable polymers such as amide polymers, ester polymers, (3) urethane polymers, and urea polymers obtained by condensation. Among these, acrylic polymers, methacrylic polymers, and acrylic-methacrylic copolymers can be preferably used. Since these polymers have less specific adsorption at the time of immobilizing proteins such as antibodies, background signals can be suppressed in antibody chips and the like, so that highly sensitive measurement is possible. The azo compound-based photoresponsive component such as the formulas (1) to (3) is preferably used as a double bond monomer as a homopolymer or a copolymer with a double bond monomer such as MMA.

  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 target molecule 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 caused by the presence of a light-responsive component in the matrix material and, for example, induced by a change in the volume, density, free volume, etc. of the matrix material or the light-responsive component upon light irradiation. it is conceivable that.

  The three-dimensional form of the solid phase material 10 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. In the case where the solid phase material 10 has a planar surface such as a film shape, a sheet shape, or a flat plate shape, the selective adsorption region is arranged in an array shape by supplying and adsorbing target molecules to the surface, for example, in an array shape. And so on.

  In addition, as a photoresponsive component or a resin or a resin composition containing the photoresponsive component that can be used in the present invention, JP-A Nos. 2003-329682, 2004-93996, and 2004-251801 are known. The carrier and the light immobilization material described in the publication 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 51998 are incorporated by reference.

(Target molecule)
The target molecule is not particularly limited, and examples thereof include inorganic materials, organic material groups, biomaterials, biomaterials and biomaterials, and composite materials obtained by combining these two types. Examples of the inorganic material include at least metals, metal oxides, semiconductors, ceramics, and glass. One type of target molecule may be used, or two or more types may be used in combination.

  Organic materials include so-called plastics. Plastic can take various forms. In addition to solid phase particles, the plastic may be in the form of particles in solution or suspension. Furthermore, the plastic particles may be single-phase particles, may be in the form of a matrix, or may be in the form of a capsule. The organic material may include an organic compound having physiological activity such as antibacterial property, anti-inflammatory property, blocking action of various biological processes, and certain agonists or antagonists. These may be low molecular compounds or high molecular compounds having one or more monomer units.

  Examples of the biomaterial include proteins, nucleic acids, saccharides, lipids, and bone forming materials, and two or more aggregates selected from these. These biomaterials include a molecule constituting the living body, a modification thereof, and a multimer thereof. The biomaterial is not limited to a material collected from a living body, and may be a material collected from a living body and subjected to artificial modification, or may be artificially synthesized. These biomaterials may be bound with a label containing a dye such as a fluorescent dye. Protein, as used herein, refers to proteins, polypeptides, and peptides of any size, structure, or function. Typically, however, the protein is at least about 6 amino acids in length. Preferably, it is at least about 10 amino acid residues long. The protein may be naturally derived, chemically synthesized, genetically engineered, or a combination of these. May be. Furthermore, the protein may contain unnatural amino acids. The protein may also be a naturally occurring protein or a fragment thereof. Examples of the protein fragment include a polypeptide obtained by digesting a full-length protein isolated from cultured cells. A protein may be a single molecule or a complex of multiple molecules. The complex includes both active and inactive proteins, and also includes aggregates having a plurality of proteins assembled in a variety of binding modes such as covalent bonds and a plurality of units. Examples of proteins and aggregates thereof include various proteins, enzymes, antigens, antibodies, lectins, and cell membrane receptors. Furthermore, compounds other than amino acid residues such as sugar chains may be bound to the protein. In the present specification, the “antibody” means an immunoglobulin produced naturally or wholly or partially synthetically. Also included are all derivatives thereof that retain specific binding ability. The term also includes any protein having a binding domain that is homologous or highly homologous to an immunoglobulin binding domain (including chimeric and humanized antibodies). The antibody may be a monoclonal antibody or a polyclonal antibody. The antibody can be a member of any immunoglobulin class, including any class (IgG, IgM, IgA, IgD and IgE). IgG class derivatives are preferred in the present invention. In addition, nucleic acids include artificial nucleic acids in addition to DNA and RNA having a single strand, two or more strands, or a portion of a single strand. Examples of the nucleic acid include genomic DNA and its microsatellite portion or a fragment containing at least a part of any gene, cDNA and its fragment, mRNA and its fragment, and DNA containing a mutation site such as a single nucleotide polymorphism site (cDNA And a DNA fragment containing a restriction enzyme site. The nucleic acid may be an oligonucleotide or a polynucleotide. Examples of the saccharide include monosaccharides, oligosaccharides, and polysaccharides. Sugars include sugar chains that are bound to proteins and lipids in vivo and some of them. Examples of the lipid include a lipid chain in lipoprotein, lipopolysaccharide or the like or a part thereof. Examples of the bone forming material include inorganic compounds such as various phosphates of calcium.

  Biological materials include at least various types of biological cells and parts thereof, tissues and organisms. Any of these may be in a living state or not in a living state. Biological cells include any animal cell of mammals and non-mammals, any cell of a plant body, cells of various organisms such as fungi, yeast, bacteria, and viruses. Furthermore, a part of the biological cell includes various organelles existing in these biological cells, as well as a membrane of the biological cell and a part of the organelle. A tissue is an aggregate of biological cells and includes a part of an animal and various tissues (including skin) of an animal and a part of a plant and an organ. Furthermore, the organism means one unit that can be propagated or propagated as an organism. Examples of organisms include fertilized eggs, microbial cells, and viruses. These biological materials may be artificially modified such as genetic engineering or chemical.

  In the present invention, it is preferable to use a biomaterial or a biomaterial as the target molecule. This is because a selective adsorption region with high selectivity can be formed according to the present invention, and adsorption of such a biological material or biological material in an active state is possible. Examples of such target molecules include various proteins, antibodies, enzymes, antigen proteins, receptor proteins and the like. Moreover, sugar chains, polynucleotides, organelles, bacteria, viruses, and aggregates of two or more of these may be mentioned.

  The target molecule preferably has an average maximum dimension (typically measured as a particle diameter such as a diameter) of 1 mm or less, more preferably 100 μm or less, further preferably 50 μm or less, and particularly preferably 10 μm or less. Can be of size. The target molecule is preferably 1 nm or more in size.

  In the adsorption step, the target molecule is supplied to the surface of the solid phase material to cause an interaction with the target molecule on the surface to adsorb the target molecule on the surface. The adsorption step is performed so that a selective adsorption region can be formed in the region after the target molecules are removed on the surface of the solid phase material. The preferred interaction has already been explained. Examples of means capable of causing such an interaction to adsorb the target molecule on the surface of the solid phase material include light irradiation and heating.

  Prior to the development of the interaction, target molecules are supplied to or near the surface of the solid phase material. Although the supply form of the target molecule is not particularly limited, it is preferable to supply the target molecule via a liquid medium. More specifically, it is preferable to apply in a state where the target molecule is dissolved or suspended in a liquid medium. When a liquid medium is used, the target molecule can be easily developed on the surface of the solid phase material, and the structure of the target molecule (eg, its secondary structure when the target molecule is a polypeptide) is maintained. This is because it can be fixed. From this point of view, the liquid medium is preferably one that can maintain the structure of the target molecule.

  By bringing the target-containing liquid into contact with the surface of the solid phase material, the target molecules are supplied to the surface of the solid phase material. In the present invention, since a solid phase material prepared in advance is used, target molecules can be supplied to a selected region on the surface of the solid phase material. For example, when the solid phase material has a planar surface such as a sheet shape, a film shape, a flat plate shape, etc., a dot shape, a stream shape, any other aspect other than these, and an aspect in which these are combined Thus, one or more target molecules can be supplied. By doing so, the selective adsorption region can be formed only in the selected region of the solid phase material surface. In addition, a plurality of selective adsorption regions can be formed in an array or the like for one type of target molecule, and one or two or more selective adsorption regions can be arrayed for each of two or more types of target molecules. It can also be formed in an array. In order to supply the target molecules in such a manner, a known liquid supply method such as an ink jet method or a pin method can be used.

  In order to cause an interaction between the surface of the solid phase material and the target molecule, it is preferable to use a material containing a photoresponsive material as the solid phase material and perform light irradiation. By light irradiation, it is considered that a complementary interaction accompanied by deformation and / or property change of the solid phase material can be caused between the surface of the solid phase material and the target molecule, and selective adsorption with high molecular recognition ability. This is because a region can be formed. In addition, an interaction is generated between the target molecule and the surface of the solid phase material under extremely relaxed conditions, and the target molecule is adsorbed (light-fixed) on the solid phase material. Moreover, according to the light fixation, it is easy to remove the target molecule after the fixation. Furthermore, the target molecule can be adsorbed by causing an interaction only in the region irradiated with light.

  Adsorption by light irradiation is achieved in the presence of a liquid medium in addition to a dry state. As described above, in order to maintain and adsorb the three-dimensional structure of the target molecule, it is preferable to irradiate with light in the presence of a liquid medium. Examples of such a liquid medium include (1) water, (2) a mixture of water and an organic solvent, (3) one or two selected from salts (including buffer salts), acids, alkalis, and surfactants. A mixed solution of a seed or more and water and / or an organic solvent can be appropriately used. The liquid medium is preferably water or water as a main medium. The component added to the liquid medium is appropriately selected according to the type of the target molecule, but one that enhances the stability of the target molecule and the interaction between the target molecule and the solid phase material is selected. In the liquid medium, for example, pH, electrolyte concentration, polarity and the like are adjusted. As the organic solvent, an aqueous solvent and a non-aqueous solvent which are organic solvents compatible with water can be used alone or in combination. When the target molecule is a biological material or biological material, it is preferable that the target molecule is sufficiently relaxed to maintain its biological activity. That is, the pH can be about 4.5 to 7.5 or less, and the salt concentration can be about the physiological saline. Such a liquid medium can also be used for supplying target molecules to a solid phase material prior to light irradiation.

The target molecule can be immobilized on the surface of the solid phase material by irradiating the target molecule on or near the surface of the solid phase material with light. The light irradiation method for fixing the light is not particularly limited. What is necessary is just to irradiate so that arbitrary lights, such as various propagation light, near field light, or evanescent light, may arrive at the surface of the solid-phase material in which a target molecule exists, or its vicinity. The wavelength range used for light fixation may be a wavelength range that causes a change in molecular structure or molecular arrangement in the photoresponsive component. Information about such wavelength ranges can be easily obtained for various available photoresponsive components or can be confirmed upon use. Typically, the wavelength is about 400 nm to 700 nm, the light intensity is about 1 to 100 mW / cm ^{2 for} about 1 minute to 240 minutes, and the operation can be performed near room temperature of about 20 ° C. to 40 ° C. Further, the adsorption region can be selected by selectively performing light irradiation on a part of the solid phase material using a known method.

  The light irradiation can be performed on the entire surface of the solid phase material, or can be performed only on a selected region. When the entire solid phase material surface is irradiated with light, target molecules on or near the solid phase material surface are immobilized on the solid phase material surface. As already described, when the target molecule is supplied only to a selected region (for example, dot shape) on the surface of the solid phase material, the target molecule is immobilized only in the region. On the other hand, when only a selected region on the surface of the solid phase material is irradiated with light, the target molecule is immobilized only in the light irradiation region. Therefore, even if the target molecule is supplied to the entire surface of the solid phase material, the target molecule is immobilized only in the region irradiated with light. As described above, by selectively supplying the target molecule to the surface of the solid phase material prepared in advance and / or selectively irradiating with light, one or more selections are made on the selected region of the surface of the solid phase material. The target adsorption region can be easily formed.

  In addition, about the light irradiation for light fixation, employ | adopt the irradiation light and the light irradiation method already described in Unexamined-Japanese-Patent No. 2003-329682, Unexamined-Japanese-Patent No. 2004-93996, and Unexamined-Japanese-Patent No. 2004-251801. Can do. The light immobilization is disclosed by the present applicant in Japanese Patent Application Laid-Open Nos. 2003-329682, 2004-93996, and 2004-251801, and these methods are used for light fixation in the present invention. Can also be applied.

  By performing the adsorption step, the target molecules are adsorbed on the surface of the solid phase material. As the target molecules are adsorbed on the surface of the solid phase material, a region where deformation or property change based on the interaction is recorded is formed on the target molecule adsorption surface of the solid phase material. When target molecules are adsorbed on the surface of a solid phase material by light irradiation, molecular recognition is performed under a relaxed condition and a region where the deformation and / or property change of the surface of the solid phase material is recorded by the interaction based on light irradiation. It can be formed to have high performance. In particular, light irradiation can be accompanied by deformation of the surface of the solid phase material, so that a gap complementary to a part of the target molecule can be easily formed in the target molecule adsorption region.

  Further, the interaction occurring in the target molecule adsorption region is maintained in the subsequent target molecule removal step. When target molecules are adsorbed to the solid phase material by light irradiation, these interactions can only cause heating above the glass transition point for the solid phase material, or the interaction that occurs in the solid phase material can be relaxed or eliminated. This is maintained unless light irradiation is performed and other target molecules are supplied to the solid phase material to fix the light and cause a new interaction. Whether target molecules have been adsorbed is determined by the fact that target molecules remain on the surface of the solid-phase material when unadsorbed target molecules are removed by washing with an appropriate liquid such as the liquid medium used during adsorption. It can be confirmed with.

(Removal process)
In the present invention, following the step of causing the target molecule to interact with the surface of the solid phase material, the target molecule is removed. By removing the target molecule from the surface of the solid phase material, the region where the target molecule is removed after adsorption can be used as a region where the target molecule can be selectively adsorbed. FIG. 1B shows a state in which the target molecules are removed by washing after the target molecules are adsorbed (light-fixed) on the surface of the solid phase material by light irradiation.

  In the removal process, it is not necessary to maintain the physical properties of the target molecule that has been adsorbed or immobilized, but the target molecule is maintained within a range that maintains at least part of the change in the area where the interaction with the target molecule is recorded (adsorption area). Try to remove it. In the adsorption process of the present invention, the target molecule is adsorbed on the surface of the solid phase material prepared in advance, and the solid phase material is not molded so as to surround the target molecule. And easily removed. For this reason, the recorded content of the interaction in the solid phase material is easily maintained.

  The removal conditions of the target molecule are not particularly limited, and can be appropriately set according to the type of the solid phase material and the target molecule. For example, a treatment that weakens the interaction with the solid phase material on the target molecule side, such as modification and decomposition of the target molecule. Specifically, heating, ultrasonic irradiation, or the like may be involved as long as the change recorded in the target adsorption region on the surface of the solid phase material is not lost. Another example is a method of removing the target molecule from the solid phase material by changing the structure or the like causing the interaction, for example, by decomposing the target molecule with an enzyme or the like to reduce the molecular weight. For example, when the target molecule is a protein, there is a method of decomposing and removing the target molecule with a protease.

  Further, for example, as the removing step, a target can be washed and removed by supplying a removal medium effective for removing the target molecule to the surface of the solid phase material that has adsorbed the target molecule. Examples of the removal medium include a medium having high affinity for the target molecule such as dissolving the target molecule. Such a removal medium preferably contains, for example, any one selected from an aqueous solution containing one or more selected from surfactants, salts, acids and alkalis in addition to water. The removal medium may contain an organic solvent as necessary. When the target molecule is a protein or the like, a removal medium can be prepared so as to dissolve, decompose, and denature the protein. Typically, the removal medium includes a buffer solution, a buffer solution containing a surfactant, and the like. When the target molecule is photofixed, the target molecule is easily removed by appropriately contacting with such a removal medium, and the recorded content of the interaction is maintained. On the other hand, the change in the surface of the solid phase material including the photoresponsive material caused by the light fixation is based on the light irradiation and is not easily erased.

  When the target molecule is removed from the surface of the solid phase material by performing the removing step, the adsorption region of the target molecule is exposed. The adsorption region after removing the target molecule records at least a part of a change (deformation or property change) based on the interaction when the target molecule is adsorbed. That is, the exposed adsorption region has a molecular recognition site for the target molecule. Therefore, this region can be used as a selective adsorption region for adsorbing target molecules.

  According to the manufacturing method of the present invention, the target molecule is supplied to the solid phase material prepared in advance, causes an interaction to adsorb the target molecule on the surface, and then removes the target molecule. Can be obtained. For this reason, it is possible to achieve both target molecule recognition ability and target molecule removal ease, and to absorb and remove target molecules under more relaxed conditions, so that an adsorption carrier capable of efficiently adsorbing the intended target molecule is obtained. Can do.

  Furthermore, according to the production method of the present invention, one or more selective adsorption regions of one or more kinds of target molecules can be easily formed at an arbitrary position on the surface of the solid phase material. In particular, two or more selective adsorption regions can be formed in an arbitrary form such as an array. For this reason, it is possible to easily form an adsorption carrier capable of efficiently adsorbing a large amount of target molecules or adsorbing a plurality of target molecules to specific positions on the same solid phase material surface.

  Note that the adsorption step and the removal step can be repeated two or more sets. For example, when a material having a planar surface is used as the solid phase material, the first adsorption step and the removal step are performed on the first region of the solid phase material surface, and the second adsorption step and the removal step are performed. The second region different from the first region can also be implemented. Such adsorption process and removal process on different regions on the surface of the solid phase material can be easily performed by using light irradiation in the adsorption process.

  Further, in addition to the solid phase material after the adsorption step and the removal step by light irradiation, a solid phase material that already has a selective adsorption region by light irradiation, such as a used adsorption carrier, is also produced by the production method of the present invention. It can be used as a solid phase material. When the same or different target molecules are supplied to the surface of such a solid phase material and irradiated with light, a new interaction occurs between the target molecule and the surface of the solid phase material, and at the same time. Changes in the selective adsorption region disappear. Thereby, a selective adsorption region can be newly born on the surface of the solid phase material.

(Adsorption carrier)
The adsorption carrier of the present invention is obtained by the production method of the present invention described above. As already explained, since the adsorption carrier obtained by the production method of the present invention is obtained by adsorbing and removing target molecules on the surface of a solid phase material prepared in advance, it has high selectivity and adsorption rate. Can be provided. Moreover, the selective adsorption region can be provided in any arrangement form.

  In addition, the adsorption carrier of the present invention includes a solid phase material containing a photoresponsive material, and one or more target molecules are photofixed on the surface of the solid phase material, and then the target molecules are removed. Resulting in one or more selective adsorption regions. According to the adsorption carrier of the present invention, the target molecule can be re-adsorbed by utilizing the deformation and / or property change of the surface of the solid phase material when the target molecule is photofixed. Since the interaction between the target molecule and the solid phase material during photofixation is specific, the target molecule can be efficiently adsorbed with high selectivity. In addition, since light fixation does not impair the properties of the target molecule and sufficiently firmly adsorbs the target molecule to the surface of the solid phase material, the same adsorption is performed when the target molecule is adsorbed by the adsorption carrier of the present invention. The state can be obtained. When a selective adsorption region is formed by light irradiation, the selective adsorption region that has been used so far is erased by performing another target molecule adsorption step and removal step on this surface, A selective adsorption region for another target molecule can be provided for the most effective use.

  For the target molecule, photoresponsive material, solid phase material, light irradiation (light fixation), selective adsorption region, etc. in the adsorption carrier of the present invention, the various embodiments in the production method of the present invention described above are applied as they are. can do.

  The adsorption carrier of the present invention can be provided with two or more selective adsorption regions arranged on the surface of the solid phase material. According to such an adsorption carrier, it is possible to efficiently adsorb a large amount of target molecules on one solid phase material and to adsorb two or more types of target molecules independently. In particular, it is useful for various applications when arranged on a plurality of selective adsorption regions in an array or the like on the surface of a solid phase material having a planar surface.

  The method for adsorbing the target molecules on the selective adsorption region of the adsorption carrier of the present invention is not particularly limited. What is necessary is just to supply the solution of a target molecule to the surface of a solid-phase material so that a target molecule and a solid-phase material surface can contact. The adsorption carrier of the present invention can adsorb target molecules with high selectivity and efficiency under mild conditions such as an aqueous medium. For example, a buffer solution containing a target molecule such as protein may be brought into contact with the surface of the solid phase material for several tens of minutes to several hours. In addition, since adsorption to the selective adsorption region of the target molecule is considered as an equilibrium reaction, the target molecular weight to be captured in the selective adsorption region can be controlled by the pH, ionic strength, and the type and concentration of the target molecule. it can.

  The adsorption carrier of the present invention is useful as a detection device for one kind or two or more kinds of target molecules. Since the detection device provided with the adsorption carrier of the present invention has high selectivity and adsorption rate with respect to the target molecule, it can detect the target molecule with high sensitivity. The three-dimensional form of the solid phase material in the detection device is not particularly limited. This kind of adsorption carrier may have a selective adsorption region on the surface of the particulate solid phase material, but is a solid phase material having a planar surface and a specific target molecule at a specific position on the surface. These selective adsorption regions are preferably used in an array or the like. This is because such a form enables simultaneous detection of a plurality of target molecules. When the target molecule is associated with a disease, the target molecule detection device of the present invention is useful as a diagnostic device for the disease.

  Further, the adsorption carrier of the present invention is useful as a cell growth device. In the cell growth device including the adsorption carrier of the present invention, the target molecule can be, for example, a cell to be grown and / or a material useful for the growth of the cell. When the target molecule is a cell, according to the cell growth device of the present invention, the adhesive cell can be adhered and grown without using a special cell adhesive material or treatment. Moreover, since a cell can be propagated only in the area | region in which the selective adsorption area | region was formed, and a cell can be propagated in a specific position, arbitrary culture forms can be provided to a cultured cell layer. Furthermore, when the target molecule is two or more types of cells and each has a selective adsorption region, different cells can be placed at different positions on the same solid phase material to be proliferated. Furthermore, when the target molecule is a material (peptide, protein, saccharide, etc.) necessary for cell growth (adhesion), a cell culture surface having such material can be formed. Also in the cell growth device of this embodiment, the cell growth region can be limited to a selective adsorption region.

  The three-dimensional form of the solid phase material in this type of cell growth device is not particularly limited, but if it has a planar surface, it can be used as a general cell growth plate. Moreover, you may have a selective adsorption area | region in the surface (outer surface and / or inner surface) of the solid-phase material which has a desired three-dimensional form. Such a three-dimensional adsorption carrier can be used as a scaffold when forming a cultured cell tissue having a three-dimensional structure.

  The adsorption carrier of the present invention is useful as a protein recovery device. In the protein recovery device including the adsorption carrier of the present invention, for example, the target molecule can be a predetermined protein such as an antibody, an enzyme, or an antigen protein. According to such a recovery device, the protein that is the target molecule can be efficiently recovered with high selectivity to the target molecule. The three-dimensional form of the solid phase material in the protein recovery device is not particularly limited. However, in view of the recovery efficiency and the like, a form that has a large surface area such as a particle and is easy to apply to a column or the like is preferable.

  The adsorption carrier of the present invention is useful as a microreactor. A target molecule in a microreactor provided with the adsorption carrier of the present invention can be a component such as a reaction substrate, a product, or a catalyst in the microreactor. For example, the target molecule can be a substrate such as an enzyme or a product in addition to a protein such as an enzyme or an antibody. According to such a microreactor, one or more components of the microreactor can be selectively adsorbed at a desired position and a specific function can be easily imparted to a specific site, so that the reaction proceeds efficiently. Can be made. The three-dimensional form of the solid phase material in the microreactor is not particularly limited, but it is preferable to have a planar surface in consideration of the formation of the reaction channel. Further, when one component of such a microreactor is constituted by the adsorption carrier of the present invention, it may be in a three-dimensional form capable of expressing the function.

  From the above, according to the present invention, the target molecule adsorption method comprising the step of supplying the target molecule to the surface of the solid phase material of the adsorption carrier of the present invention and causing the selective adsorption region to adsorb the target molecule ( A method of using the carrier for adsorption). The adsorption method of the present invention is useful as various methods according to various aspects of the adsorption carrier of the present invention. That is, the adsorption method of the present invention includes a method for using a detection device (including a diagnostic method) (a method for detecting a target molecule), a method for using a cell proliferation device (a cell proliferation method, a method for producing a cell proliferation body), and a recovery device. It is also useful as a method of using (Protein recovery method), a method of using a microreactor (Reaction method, reaction product production method),

  In addition, according to the present invention, there is also provided a solid phase body on which target molecules are adsorbed, which is obtained by adsorbing target molecules on the selective adsorption region of the adsorption carrier of the present invention. While this solid phase adsorbs target molecules with high selectivity and efficiency, the target molecules can be easily removed by washing or the like. Furthermore, a selective adsorption region capable of selectively adsorbing a new target molecule on the surface of the solid phase material can be born.

(Target molecule adsorption carrier material)
The target molecule adsorption carrier material of the present invention can include a solid phase material including a photoresponsive material. According to the material for an adsorption carrier of the present invention, a selective adsorption region for selectively adsorbing the target molecule is obtained by supplying the target molecule to the surface of the solid phase material, adsorbing it by light irradiation, and then removing it. An adsorption carrier provided can be produced. Various embodiments in the production method of the present invention described above can be applied as they are to the photoresponsive material and the solid phase material in such an adsorbent carrier material.

  EXAMPLES Hereinafter, although this invention is demonstrated more concretely based on an Example, this invention is not limited to a following example.

(Preparation of solid phase material)
Using a polymethacrylate polymer compound (m: n = 15: 85) represented by the following formula, a thin film having a thickness of about 40 nm is produced on a glass substrate, and the film surface is used as a solid phase material surface. The solid phase material was prepared.

(Adsorption of protein to azopolymer slide (immobilization))
Proteins of different sizes ((1) ribonuclease A, (2) antibody (IgG), (3) bovine serum albumin (BSA) or human serum albumin (HSA), (4) lysozyme, (5) catalase, (6) ferritin , (7) thyroglobulin, see FIG. 3) was diluted with 0.01% Tween20-containing phosphate buffer (TPBS) to a concentration of 10 μg / mL on an azopolymer slide according to the predetermined layout shown in FIG. did. After water was distilled off under reduced pressure, light was irradiated with Lumifix (maximum wavelength: 470 nm, 20 mW / cm ² ) for 30 minutes to adsorb (immobilize) the protein onto the azopolymer slide. Then, the slide was immersed in TPBS and stirred and washed for 5 minutes three times to remove excess protein, and an azopolymer slide on which various proteins were adsorbed was obtained (see FIG. 2 (a)).

(Removal of protein)
The slide on which the protein was adsorbed was put into 2 wt% sodium dodecyl sulfate phosphate buffer (SDS / PBS), which was being stirred, and washed with stirring. After washing for a certain time, the slide was taken out, washed with water and dried (see FIG. 2B).

(Evaluation of protein re-adsorption ability)
As shown in FIG. 2 (c), the TPBS solution (10 μg / ml) of the antibody is placed on the half of the slide after removing the protein (the lower half of the slide shown in FIG. 4) using a gap cover glass. Let stand for hours. (See FIG. 2 (c)). After carefully removing the gap cover glass in the PBS solution, it was immersed in a TPBS solution and washed twice with stirring for 1 minute. Subsequently, the gelatin solution of the antibody labeled with Cy5 was placed on the entire surface of the azopolymer slide using a gap cover glass, and the antibody on the slide was immunoreacted to carry out fluorescence labeling. (See FIG. 2 (d)).

  After reacting for 30 minutes, the azopolymer slide was immersed in TPBS and stirred and washed for 1 minute twice. Thereafter, the fluorescence on the slide glass was measured with a commercially available array scanner. The results are shown in FIG.

  As shown in FIG. 5, IgG that could not be removed by washing was present in the region where IgG was removed after light fixation. However, it was revealed that more IgG was re-adsorbed than other protein parts by re-adsorption. In FIGS. 4 and 5, “8” is a portion spotted with a solution containing no protein, and the protein is not immobilized, but no re-adsorption to this portion has occurred. From this, it was clarified that the affinity with IgG does not occur only by contacting with the solution, and only the region where the IgG is removed after photofixation has affinity with IgG and is re-adsorbed.

(Evaluation of protein re-adsorption and selectivity)
In this example, proteins of various sizes were spotted in the same manner as in Example 1, light-irradiated by light irradiation, and removed. Two azopolymer slides were prepared in the same manner as these steps. The slides were designated as slide A and slide B, respectively. A TPBS solution (10 μg / ml) of IgG, which is one of the immobilized proteins, was placed on the lower half area of Slide A. In addition, a TPBS solution (10 μg / ml) of HSA, which is another protein, was placed on the lower half region of slide B. These slides A and B were each allowed to stand for 2 hours.

  After carefully removing the gap cover glass in the TPBS solution, it was immersed in the TPBS solution and washed twice with stirring for 1 minute. Subsequently, a gelatin solution of Cy5-labeled antiIgG antibody was placed on the entire surface of slide A, and fluorescently labeled by immunoreacting with IgG on the slide. In addition, a gelatin solution of Cy5-labeled antiHSA antibody was placed on the entire surface of slide B, and fluorescently labeled by immunoreacting with HSA on the slide. After reacting both slides A and B for 30 minutes, the azopolymer slide was immersed in TPBS and stirred and washed twice for 1 minute. Thereafter, the fluorescence on the slide glass was measured with a commercially available array scanner. The results are shown in FIGS.

  FIG. 6 shows the result of slide A. In the reaction with the anti-IgG antibody, effective re-adsorption to the region to which IgG was adsorbed (indicated by “2” in FIGS. 4 and 6) was shown. FIG. 7 shows the result of slide B. In the reaction with the antiHSA antibody, effective re-adsorption to the region where the HSA was adsorbed (indicated by “3” in FIGS. 4 and 7) was shown. Further, when FIGS. 6 and 7 are compared, no re-adsorption of HSA to the region where IgG was adsorbed, and similarly no re-adsorption of IgG to the region where HSA was adsorbed.

  From the above, it was found that each of the IgG and HSA was specifically re-adsorbed at the portion where the IgG and HSA were adsorbed and removed. From these, it can be seen that there is a region that can be adsorbed on the surface in the region where these proteins are adsorbed and removed. According to the present inventors, it is known that when a micro object is light-immobilized with a photoresponsive material and then removed, a void that follows a part of the shape of the microobject is formed on the surface of the photoresponsive material. . Therefore, it was considered that voids were formed following the shape of part of the protein in the region where the protein was photofixed and then removed. In addition, since the region where IgG is immobilized is only IgG and the region where HSA is immobilized is resorbed only HSA, the region formed by this method may have a property of selectively adsorbing proteins. It was revealed.

(Effects of template concentration and resorbed protein concentration)
In this example, slides with a protein concentration of 1 μg / mL in Example 1 were prepared, and the amount of re-adsorption was evaluated through the same steps as in Example 2. The resorbed protein concentration is 10 μg / mL. The results are shown in FIGS. The amount of re-adsorption was evaluated in the same manner as in Examples 1 and 2, except that the protein concentration for immobilization was 10 μg / ml and the re-adsorption protein concentration was 1 μg / ml. These results are shown in FIGS.

  FIG. 8 shows the result of slide A, and it was revealed that IgG was effectively re-adsorbed to the IgG adsorption region. FIG. 9 shows the result of slide B. It was revealed that HSA was effectively re-adsorbed to the HSA adsorption region. Thus, it was revealed that re-adsorption occurred as in the case of 10 μg / mL even when the protein concentration during adsorption was low (1 μg / ml).

  FIG. 10 shows the result of slide A. No significant re-adsorption of IgG was observed in the IgG adsorption region. FIG. 11 shows the result of slide B. No significant resorption of HSA was observed in the HSA adsorption region.

  From these results, it was suggested that the re-adsorption process is not irreversible but proceeds in an equilibrium process.

  In the above examples, the preferred embodiments relating to the method for providing a solid phase material having a region that selectively adsorbs the target molecule on the surface of the present invention and the recognition ability evaluation have been described. And the manufacturing method is not limited to the said Example.

Claims (19)

A method for producing an adsorption carrier having one or more selective adsorption regions of one or more kinds of target molecules,
The following steps:
(A) supplying a target molecule to the surface of a solid phase material to cause interaction with the target molecule on the surface and adsorbing to the surface;
(B) removing the target molecule from the surface of the solid phase material;
A manufacturing method comprising:   2. The manufacturing method according to claim 1, wherein the selective adsorption region has at least a part of deformation and / or changed properties of the solid-phase material obtained by adsorption of the target molecule.   The manufacturing method according to claim 1, wherein the selective adsorption region has a gap complementary to a part of the target molecule.   The said (a) process is a process in any one of Claims 1-3 which is a process including fixing the said target molecule to the surface of the said solid-phase material containing a photoresponsive material by light irradiation. Method.   The manufacturing method according to claim 4, wherein the photoresponsive material includes a polymer material.   The manufacturing method according to claim 4, wherein the photoresponsive material contains azobenzene or an azobenzene derivative as a photoresponsive component.   The production method according to claim 6, wherein the azobenzene derivative is push-pull azobenzene or aminoazobenzene.   In the step (b), a surface of the solid phase material is provided with a removal medium containing any one selected from water and an aqueous solution containing one or more selected from surfactants, salts, acids and alkalis. The manufacturing method according to claim 1, which is a step of supplying and removing the target molecule.   The manufacturing method according to claim 1, wherein an average size of the target molecules is 1 nm or more and 100 μm or less.   The manufacturing method according to claim 1, wherein the target molecule is a biological material or a biological material.   The step (a) is a step of causing the target molecule to be adsorbed by causing an interaction with one or more types of the target molecules in one or more selected regions of the surface of the solid phase material. The manufacturing method in any one of Claims 1-10.   A solid carrier for adsorption of target molecules, obtained by the production method according to claim 1. A solid phase material containing a photoresponsive material;
One or more selective adsorption regions obtained by photofixing one or more target molecules on the surface of the solid phase material and then removing the target molecules;
A carrier for adsorbing target molecules.   The adsorption carrier according to claim 13, comprising two or more selective adsorption regions arranged on the surface of the solid phase material.   A target molecule detection device comprising the adsorption carrier according to claim 12.   A cell growth device comprising the adsorption carrier according to any one of claims 12 to 14.   A protein recovery device comprising the adsorption carrier according to claim 12.   A microreactor comprising the adsorption carrier according to any one of claims 12 to 14.   A target molecule adsorption carrier material comprising a solid phase material containing a photoresponsive material.