JP2009109213A - Microdevice utilizing functional particle, and processing method using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microdevice suitable for processing such as separation, immobilization, analysis, extraction, purification or reaction of a target material. <P>SOLUTION: The microdevice having particles bondable with the target material and a member provided with a plurality of hollows has characteristics wherein a material or a functional group bondable with the target material is immobilized on the surface of a particle body, and the density of the particle is 3.5-23 g/cm<SP>3</SP>, and the plurality of hollows can store the particles. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a microdevice suitable for processing such as separation, immobilization, analysis, extraction, purification or reaction of a target substance. The present invention also relates to a method for treating a target substance using such particles.

Analyzes are performed by binding substances such as nucleic acids, proteins, sugar chains or cells to particles, and binding, adsorbing or reacting the substances on these particles with target substances such as bases, nucleic acids, proteins, antigens or sugar chains. Various systems for performing extraction, purification or reaction have been proposed. For example, there has been proposed a technique for performing hybridization or sequencing of nucleic acids by arranging microbeads bound with nucleic acids or the like or putting them in a microcell. However, many of the microbeads that are usually used are made of a material with a low specific gravity such as a resin base material or silica, and it is difficult for the microbeads to enter the microcell, or when the sample containing the target substance flows, There was a problem that it was easy to move or jump out of the microcell. Therefore, when the above microbeads are used, an additional step of centrifuging is added to forcibly enter the microcell, or the flow of the sample containing the target substance is suppressed (ie, the flow rate of the sample to be supplied). Cell shape (for example, a deeper cell or a cell side wall surface with a steeper inclination) that suppresses the movement / jumping of the microbeads at the expense of the flow must be employed.
Nature Biotechnology, Vol.18, P.630-634 (2000) Special Table 2007-523627

  The present invention has been made in view of the above circumstances. That is, an object of the present invention is to provide a device in which the above-described problem when flowing a sample or the like is at least reduced. More specifically, the problem of the present invention is that the movement of particles is suppressed, and the target substance is separated, immobilized, analyzed, extracted, and purified on each particle by fixing the particle at a substantially constant position. Alternatively, it is to provide a device that enables reaction and the like.

In order to solve the above problems, the present invention provides:
A microdevice comprising particles capable of binding a target substance and a member provided with a plurality of depressions,
A substance or functional group capable of binding a target substance is immobilized on the surface of the particle body of the particle, the density of the particle is 3.5 g / cm ^{3 to} 23 g / cm ³ , and the plurality of depressions are particles A microdevice is provided that is configured to be capable of accommodating a battery.

As used herein, “micro” of “micro device” means that the size of a structure such as a depression is on the order of micrometers (about 1 to 10 ³ μm), or “a member provided with a plurality of depressions”. It means substantially in the order of millimeters (about 1 to 10 ² mm).

  The particle used in the microdevice of the present invention has a “substance or functional group capable of binding a target substance” immobilized on its surface. In other words, the “substance or functional group that binds to the target substance” is immobilized. Therefore, when the target substance and the particle coexist, the target substance can bind to the particle, so that the target substance can be used not only for various applications such as separation, purification or extraction of the target substance, but also tailor-made. Particles can also be used for medical technology applications. Here, the “target substance” substantially means a substance that can be subjected not only to separation but also to various treatments such as extraction, quantification, purification or analysis, and is directly or indirectly applied to particles. Any kind of substance can be used as long as it can bind. Specific target substances include, for example, nucleic acids, proteins (including avidin and biotinylated HRP, etc.), sugars, lipids, peptides, cells, fungi, bacteria, yeasts, viruses, glycolipids, glycoproteins, complexes, inorganic substances, A vector, a low molecular compound, a high molecular compound, an antibody, an antigen, etc. can be mentioned. The particles used in the microdevice of the present invention can be said to exhibit various functions in that they can be used for separation, purification, extraction or analysis of various target substances. Therefore, the particles used in the microdevice of the present invention can be called “functional particles”.

Particles are utilized in the micro device of the present invention, density of ^{3.5g / cm 3 ~23g / cm 3} , that the density (or specific gravity) is greater than the commonly particles used in the separation of a target substance It has characteristics. Moreover, the particle | grains utilized for the microdevice of this invention also have the characteristics that the through-hole is not formed in the particle | grain main body and it is not a porous form. Therefore, in general, the specific surface area of the particles is relatively small as 0.0005m ² / g~10m ² / g. In the present specification, “the particle body does not have a through-hole” means that the particle body is substantially solid and the particle does not have an internal through-network structure. That is, “the particle body does not have a through-hole” in the present specification means “the particle body or the particle body core is solid”, “even if the particle surface is uneven, the recess is not formed. It is synonymous with “it does not exist even inside the particle” and “the bulk density is larger when compared with general porous particles”.

The present invention also provides a processing method for analyzing, extracting, purifying, or reacting a target substance using the above-described microdevice. Such a processing method of the present invention includes:
(I) supplying a liquid containing the above-mentioned particles or a liquid in which the above-mentioned particles are dispersed to a “member provided with a plurality of depressions”, and storing one particle in each of the plurality of depressions; And (ii) supplying a sample containing the target substance to the surface of the “member provided with a plurality of depressions”, and bringing the sample and particles into contact with each other. This method is characterized in that in step (i), one particle is accommodated in each of the depressions by allowing the particles contained in the liquid to naturally settle.

  In the present specification, “sample comprising target substance” means nucleic acid, protein, sugar, lipid, peptide, cell, fungus, bacteria, yeast, virus, glycolipid, glycoprotein, complex, inorganic substance, vector, Samples containing target substances such as low molecular weight compounds, high molecular weight compounds, antibodies or antigens (for example, body fluids such as human or animal urine, blood, serum, plasma, semen, saliva, sweat, tears, ascites, amniotic fluid; human Or suspensions, extracts, lysates or lysates of animal organs, hair, skin, mucous membranes, nails, bones, muscles or nerve tissues; stool suspensions, extracts, lysates or lysates; cultured cells Or suspension, extract, lysate or disruption of cultured tissue; virus suspension, extract, lysate or disruption; cell suspension, extract, lysis or disruption; soil suspension Suspension, extract, lysate or crushed liquid; plant suspension, extract, lysate or crushed liquid ; Are substantially means that the drainage and the like); food and processed food suspension, extract, lysate or homogenate. Further, in this specification, “natural sedimentation” substantially means that particles are settled in a liquid such as a sample under the action of gravity.

Particles are utilized in the micro device of the present invention, since the density is as large as ^{3.5g / cm 3 ~23g / cm 3} , it recess have been subjected member surface (e.g., "well-well plate member") is If there is a liquid such as water in the surroundings, the density difference between the surroundings (that is, the density difference between the liquid and the particles) is large, and the particles are caused by gravity (especially natural sedimentation). Will enter the depression, and once entering the depression, there is an effect that the particles are difficult to jump out of the depression.

  As described above, the particles used in the microdevice of the present invention are likely to enter the depression due to natural sedimentation, and once entering the depression, the particles are less likely to jump out of the depression. Unlike low-density particles consisting of, etc., the degree of freedom of flow of the sample such as centrifuge is increased (for example, “the flow rate of the sample when contacting the sample and the particle can be increased”), and Reducing the inclination of the side wall surface inside the depression (specifically, “decreasing the inclination of the side wall surface inside the depression relative to the opening surface of the depression”) or reducing the depth of the depression, etc. It is possible to adopt a recess shape that can easily or sufficiently flow into the interior of the housing.

Here, particles are utilized in micro device of the present invention, not only the density is large, the specific surface area of 0.0005m ² / g~10m ² / g and a relatively small, regarded as substantially non-porous Therefore, non-specific binding of substances other than the target substance to the particles can be suppressed. In other words, due to the non-porous nature of the particles, there are few or substantially no particle pores or particle surfaces that can absorb and adsorb substances other than the target substance, and substances other than the target substance bind to the particles. Can be suppressed. In addition, it is possible to reduce the apparent non-specific binding that the target substance entering the hole is difficult to go out.

  As described above, since the particles used in the microdevice of the present invention can suppress non-specific binding, the target substance can be efficiently purified and separated by simple operations. .

Moreover, particles used in the present invention, as described above, not only the density is large, be regarded as relatively small substantially non-porous and the specific surface area of the particles 0.0005m ² / g~10m ² / g It is a particle that can As a result, when the particles are supplied to the sample containing the target substance, a state where a gas such as air is taken into the particles is suppressed (that is, the buoyancy action of the gas can be eliminated). Therefore, when the liquid in which the particles are dispersed is supplied to the “member provided with a plurality of depressions”, the effect is that the particles are likely to enter the depressions and are difficult to jump out once entering the depressions. Furthermore, as described above, the particles used in the treatment method of the present invention are particles that can suppress “non-specific binding of a substance other than the target substance to the particle”. Therefore, in the treatment method of the present invention, even when a substance other than the target substance is contained in the sample, the target substance can be preferentially bound to the particles, and the target substance can be efficiently separated.

  Incidentally, a polymer may be deposited on the surface of the particle body of the particles used in the microdevice of the present invention. In this case, the “substance or functional group capable of binding the target substance” can be immobilized on the surface of the attached polymer (hereinafter also referred to as “attached polymer”). Thus, even if it is difficult to covalently bond the “substance or functional group capable of binding the target substance” to the particle body, the “substance or functional group capable of binding the target substance” Can be immobilized on the particle surface. In applications where the “substance or functional group capable of binding the target substance” immobilized on the surface of the particle body is likely to be separated from the surface of the particle body in applications under various conditions of use. , Separation from the particle body can be prevented by immobilizing the “substance or functional group capable of binding the target substance” to the adherend polymer. In addition, if a polymer to which various molecules or metal ions are difficult to permeate is selected as the polymer to be deposited, the elution of metal ions from the surface of the particle body or the inside of the particles (that is, metal ions that are constituent components of the particles) is suppressed. It is possible to suppress unnecessary reactions caused by metal ions and the like in various uses of particles.

  Below, the particle | grains used by this invention are demonstrated in detail first, Then, the microdevice and processing method of this invention are demonstrated.

The particles used in the microdevice of the present invention have a density suitable for processing such as separation of a target substance. That is, body fluids such as human or animal urine, blood, serum, plasma, semen, saliva, sweat, tears, ascites, amniotic fluid; human or animal organs, hair, skin, mucous membranes, nails, bones, muscles or nerve tissues, etc. Suspension, extract, lysate or disrupted solution of stool; Suspension suspension, extract, lysate or disrupted solution; Suspension, extract, lysate or disrupted solution of cultured cell or tissue; Suspended virus Suspended liquid, extract, lysate or crushed liquid; cell suspension, extract, lysate or crushed liquid; soil suspension, extract, lysate or crushed liquid; plant suspension, extract , Dissolved liquid or crushed liquid; food / processed food suspension, extract, dissolved liquid or crushed liquid; particles having such a density that the sedimentation rate of the particles becomes relatively large when dispersed in a sample such as waste water. Has. When the density of the particles is smaller than 3.5 g / cm ³ , the moving speed of the particles by only natural sedimentation is not practically preferable. On the other hand, when the density of the particles is larger than 23 g / cm ³ , “the target substance is bound. It is not preferable for the agitation performed when the “substantially possible substance or functional group” is immobilized on the particle body surface. Thus, the density of the particles are utilized in the micro device of the present invention ^{is 3.5g / cm 3 ~23g / cm 3} , more preferably from ^{3.5g / cm 3 ~9.0g / cm 3} , more ^{preferably 5.0g / cm 3 ~7.0g / cm 3} . Here, “density” in the present specification means a true density in which only the volume occupied by the substance itself is a volume for density calculation, and a true density measuring device Ultrapycnometer 1000 (manufactured by Yuasa Ionics Co., Ltd.). ) Can be used.

Preferred specific surface area of the particles that are used in micro-devices ^are 0.0005m ² / g~10m 2 / g, more preferably from 0.0005m ² / g~6m ² / g, more preferably 0. 002m a ² / ^{g~3m 2} / g, still more preferably 0.005m ² /g~1.0m ² / g, for ^example, a 0.01m ² /g~0.06m 2 / g. Therefore, the particles used in the microdevice can be regarded as substantially non-porous, and the possibility that a substance other than the target substance binds to the particles (ie, “non-specific binding possibility”) is suppressed. It has been. This means that the separation accuracy of the target substance is improved. In addition, the “specific surface area” referred to in the present specification is a specific surface area obtained by using a specific surface area pore distribution measuring device Bellsorb-mini (manufactured by Nippon Bell Co., Ltd.). The particles used in the microdevice of the present invention are preferably substantially non-porous and do not have a through-hole (that is, an internal through-network structure). This is because, when particles are supplied to a sample containing a target substance, gas such as air is prevented from being taken into the particles (that is, the buoyancy effect of gas can be eliminated). This is because, when the material is supplied to the “member provided with the dent”, the particles are likely to enter the dent, and once entering the dent, it is difficult to jump out.

The material of the particle body is not particularly limited as long as the particles have the above-described density and specific surface area. Preferably, the particle body is formed from a metal or metal oxide, for example selected from the group consisting of zirconia (zirconium oxide, yttrium-doped zirconium oxide), iron oxide, alumina, nickel, cobalt, iron, copper and aluminum. Formed of at least one material. In particular, in the case of particles having a density higher than 9.0 g / cm ³ , for example, the particle body includes Ag (silver), Au (gold), Pt (platinum), Pd (palladium), W (tungsten), At least one selected from the group consisting of Rh (rhodium), Os (osmium), Re (rhenium), Ir (iridium), Ru (ruthenium), Mo (molybdenum), Hf (hafnium) and Ta (tantalum) Or may be formed of at least one typical metal element selected from the group consisting of Pb (lead), Bi (bismuth) and Tl (thallium). Good.

  It is also effective that the particles used in the microdevice of the present invention are magnetic. (Hereinafter, magnetic particles are also called “magnetic particles”). Because, when a liquid in which particles are dispersed (for example, water) is supplied to a “member provided with a plurality of depressions”, it is possible to further suppress movement and ejection of particles once entering the depressions by a magnetic field, In addition, it is possible to induce the particles into the depressions using the magnetism, and to take out and collect the particles from the depressions. Furthermore, it is also possible to move the particles in the recesses to perform stirring, rotation, etc., to move the particles from one recess to another, or to move in the groove-shaped recess.

  In the case where an adherence polymer is provided on the surface of the particle body, it is usually difficult to impart magnetism to the adherence polymer. Therefore, it is preferable to use a particle body having magnetism.

The material of the main body of the magnetic particles is not particularly limited as long as the particles are magnetized. For example, the main body of the magnetic particles is formed of at least one iron oxide selected from the group consisting of oxides of garnet structure comprising transition metal and iron, ferrite, magnetite and γ-iron oxide. Is preferred. Alternatively, the main body of the magnetic particles may include at least one metal material selected from the group consisting of nickel, cobalt, iron, and an alloy including these metals. Here, “an oxide having a garnet structure including a transition metal and iron” is generally called YIG. For example, a compound represented by a composition formula of Y ₃ Fe ₅ O ₁₂ or this compound Bi _x Y _3-x Fe ₅ O ₁₂ (0 <X <3) in which a part of Y in the bismuth is substituted.

  Alternatively, the magnetic particles may be formed by coating non-magnetic particles with a magnetic material, or by simply depositing a magnetic material on non-magnetic particles. Also good. For coating and depositing the magnetic substance, an electroless plating method, an electroplating method, a sputtering method, a vacuum deposition method, an ion plating method, a chemical vapor deposition method, or the like can be used. Here, the “particles that are not magnetic” include, for example, high-density particles made of zirconia (zirconium oxide, yttrium-added zirconium oxide), alumina, or the like. Further, when the ratio of the high-density magnetic substance is increased, particles made of aluminum, silica, resin or the like having a lower density can be used. The “magnetic substance” used for coating or deposition is similar to the above-described magnetic particle material, such as ferrite, magnetite, γ-iron oxide, or an oxide having a garnet structure containing transition metal and iron. In addition to iron oxides, mention may also be made of nickel, cobalt, iron or alloys comprising these metals.

  When obtaining magnetic particles by coating or depositing a magnetic substance, if the amount of the magnetic substance coating formed on the particle surface is too small, the value of magnetization of the particles becomes small, which is not preferable for magnetic separation. Accordingly, the volume of the magnetic material coating is preferably 5% or more with respect to the volume of the particles (particles including the magnetic material coating). The thickness of the magnetic substance film is preferably 1.7% or more with respect to the diameter of the particles (particles including the magnetic substance film). Incidentally, not only a mode in which the magnetic material film is provided for “non-magnetic particles” but also a mode in which a magnetic material is included in the “non-magnetic particles” is conceivable.

Examples of magnetic properties of magnetic particles include “saturation magnetization” and “coercivity”. In general, the larger the saturation magnetization value, the better the responsiveness of particles to a magnetic field. Here, in order to provide magnetism to particles having a relatively large density as used in the microdevice of the present invention, it is necessary to provide a magnetic substance on the surface or inside of the particles that are not magnetized. . Here, since the density of the magnetic substance is smaller than that of the non-magnetic particles, the necessary density must be maintained by limiting the amount of the magnetic substance to be provided. In addition, when a non-magnetic polymer is deposited on the particle body, it is practically difficult to obtain a larger saturation magnetization than in the case where particles are made of only a magnetic substance. From the above, it can be said that it is actually difficult to obtain a saturation magnetization larger than 85 A · m ² / kg. On the other hand, if the saturation magnetization is less than 0.5 A · m ² / kg, the responsiveness of the particles to the magnetic field is undesirably lowered. Therefore, the saturation magnetization amount of the particles is preferably 0.5 A · m ² / kg to 85 A · m ² / kg (0.5 emu / g to 85 emu / g), more preferably 3 A · m ² / kg to a ^{10A · m 2 / kg (3emu} / g~10emu / g), for example ^{4A · m 2 / kg~7A · m} 2 / kg (4emu / g~7emu / g). In general, as the coercive force value increases, the particles tend to aggregate. However, if the coercive force is too large, the aggregating action becomes too strong and the particles are not dispersed, which is not preferable in terms of binding the target substance. Therefore, the coercive force is preferably 0 kA / m to 23 KA / m (0 to 300 oersted), more preferably 0 kA / m to 15.95 kA / m (0 to 200 oersted), and further preferably 0 kA. / M to 7.97 kA / m (0 to 100 oersted).

  The values of “saturation magnetization” and “coercive force” in this specification are values measured using a vibrating sample magnetometer (model VSM-5, manufactured by Toei Kogyo Co., Ltd.). Specifically, the value of “saturation magnetization” is a value obtained from the amount of magnetization when a magnetic field of 797 kA / m (10 kilo-Oersted) is applied. The value of “coercive force” is a value of an applied magnetic field in which the magnetization amount becomes zero when a magnetic field of 797 kA / m is applied, the magnetic field is returned to zero, and the magnetic field is gradually increased in the opposite direction. is there.

  The shape of the particles used in the microdevice of the present invention is not particularly limited, and may be, for example, a spherical shape, an ellipsoidal shape, a grain shape, a plate shape, a needle shape, a polyhedral shape (for example, a cubic shape), or the like. However, in order to reduce the variation between particles upon binding to the target substance, the particle shape is preferably a regular shape, and a spherical shape is particularly preferable. Further, when a magnetic substance film is provided on a non-magnetic particle main body, it is preferable that the “non-magnetic particle main body” has a spherical shape or an ellipsoidal shape.

  The average particle size (ie, “average particle size”) is preferably 1 μm to 1 mm. If the average particle size is smaller than 1 μm, it will be difficult to sufficiently increase the moving speed due to the natural sedimentation of the particles when the target substance is separated. On the other hand, if the average particle size is larger than 1 mm, it can of course be used. Even particles with a small specific gravity have high sedimentation properties and less advantage. This is because the particles may settle before the binding with the target substance occurs, and the target substance may not be sufficiently separated. An average particle size of 5 μm to 500 μm is more preferable, and an average particle size of 10 μm to 100 μm is still more preferable. “Particle size” as used herein refers to the length of the particle in any direction (when the polymer is applied to the particle body, the length of the particle including the thickness of the applied polymer) The maximum length is substantially meant, and “average particle size” (ie, “average particle size”) refers to, for example, 300 particles based on electron or optical micrographs of the particles. The size is measured and the particle size calculated as the number average is substantially meant. It should be noted that when the size of the particles made of pure metal is small, rapid oxidation is likely to occur, and in some cases, there is a risk that the particles may ignite, but a relatively large particle size as used in the microdevice of the present invention. Then, rapid oxidation is unlikely to occur, and the risk of ignition of particles is reduced.

  The “substance capable of binding the target substance” (hereinafter also referred to as “substance to which the target substance can bind”) immobilized on the surface of the particle body is composed of biotin, avidin, streptavidin, and neutravidin. It is preferably at least one substance selected from the group. The “functional group capable of binding the target substance” (hereinafter also referred to as “functional group to which the target substance can bind”) immobilized on the surface of the particle main body is a carboxyl group, a hydroxyl group, or an epoxy group. , Tosyl groups, succinimide groups, maleimide groups, thiol groups, thioether groups, disulfide groups and other sulfide functional groups, aldehyde groups, azide groups, hydrazide groups, primary amino groups, secondary amino groups, tertiary amino groups, imide esters It is preferably at least one functional group selected from the group consisting of a group, a carbodiimide group, an isocyanate group, an iodoacetyl group, a halogen-substituted product of a carboxyl group, and a double bond. The “functional group to which the target substance can bind” may be a derivative of the above-described functional group.

  In the present specification, the term “immobilization” generally refers to an embodiment in which a “substance that can bind to a target substance” or a “functional group that can bind to a target substance” exists in the vicinity of the surface of the particle body. It does not necessarily mean only the mode in which the “substance capable of binding the target substance” or the “functional group to which the target substance can bind” is directly attached to the surface of the particle body. In addition, “immobilization” substantially means an embodiment in which “substance or functional group capable of binding to a target substance” is immobilized on at least a part of the particle surface. The “substance or functional group” does not necessarily have to be immobilized over the entire particle surface. However, in a preferred embodiment, the “substance or functional group to which the target substance can bind” exists over the entire particle surface so that the particle body is included in the “substance or functional group to which the target substance can bind”. In the present specification, the term “target substance binds” not only includes an aspect in which the target substance is “adsorbed” or “absorbed” with respect to the particle, but also between the target substance and the particle. It also encompasses embodiments in which the target substance is bound to the particle due to the various “affinities” that work.

  Since the “substance that can bind to the target substance” or “functional group that can bind to the target substance” is immobilized on the particle body of the particle, the target substance can be bound to the particle via the substance or functional group. Can do.

  The method for immobilizing the “substance that can bind to the target substance” to the particle body is not particularly limited, and any technique that can bind or attach the “substance that can bind to the target substance” to the particle body. Any method may be used. It is not limited to directly bonding or attaching the “substance to which the target substance can bind” to the particle body, but if necessary, a silicon-containing substance (for example, siloxane, silane coupling agent and sodium silicate), or Another substance such as a resin having a functional group to which the target substance can bind or adhere is previously attached or introduced to the particle body, or the surface of the particle body is subjected to noble metal deposition treatment and then the target substance is bound or By attaching or introducing another substance such as a sulfur-containing compound having a functional group capable of attaching to the particle body in advance, the “substance to which the target substance can bind” may be easily immobilized on the particle. When a silicon-containing substance is used, a “substance that can bind to the target substance” is immobilized on the surface of the particle body, and the silicon-containing substance is present.

An example of a technique for immobilizing a “substance that can bind to a target substance” on a particle body is described by, for example, reacting a silane coupling agent having an epoxy group or an amino group on the surface of the particle body with a “target substance”. Can be immobilized on the particle.

  Similarly, the method for immobilizing the “functional group to which the target substance can bind” to the particle body is not particularly limited, and the “functional group to which the target substance can bind” may be bonded or attached to the particle. Any method may be used as long as it is possible. For example, if necessary, the “functional group to which the target substance can bind” may be chemically treated and converted into another functional group to change the reactivity, adsorptivity, and the like. Similar to the “substance to which the target substance can bind”, not only the “functional group to which the target substance can bind” is directly bonded or attached to the particle body, but also a silicon-containing substance (for example, siloxane, Silane coupling agent and sodium silicate, etc.), other substances such as resins having functional groups to which the target substance can bind or adhere are previously attached to or introduced into the particle body, or noble metal is deposited on the surface of the particle body By attaching or introducing another substance such as a sulfur-containing compound having a functional group to which the target substance can be bonded or attached after the treatment to the particle body, the “functional group to which the target substance can bind” May be easily fixed to the particle body. For example, when a silicon-containing substance is used, the “functional group to which the target substance can bind” is immobilized on the surface of the particle body, and the silicon-containing substance is present.

  Hereinafter, a technique using siloxane will be described as an example of a technique for immobilizing a “functional group to which a target substance can bind” to particles.

<< Immobilization of functional groups using siloxane >>
This method is a method in which the surface of the particle body is previously coated with 1,3,5,7-tetramethylcyclotetrasiloxane (hereinafter abbreviated as “TMCTS”). Such a method has a feature that the reaction is terminated when only one layer of the TMCTS film is formed on the surface of the particle body.

(Pretreatment process)
First, TMCTS is added to a dispersion obtained by dispersing precursor particles in an organic solvent. At this time, a sufficient amount of TMCTS is added to form one layer of TMCTS on the particle surface. Next, the dispersion is evaporated to remove the solvent from the dispersion, and the particles are heated and dried in a vacuum desiccator, and then heated in a thermostatic bath at 150 ° C. The organic solvent to be used may be any kind of organic solvent as long as it is easily evaporated by an evaporator and has a low boiling point, and examples thereof include toluene, hexane, and benzene. If the inside of the vacuum desiccator is heated and dried, the same effect as the chemical vapor deposition method (CVD method) is brought about. Therefore, the heating temperature needs to be in a range where TMCTS evaporates and decomposition does not occur. . Specifically, a heating temperature of about 30 to 80 ° C. is preferable. The heating process in the thermostat performed at the end is a process of advancing the reaction between TMCTS attached to the particle surfaces. If the temperature becomes too high, TMCTS is likely to be decomposed, and if it is carried out for a long time, there will be no reactive sites required in the subsequent functional group immobilization step, so a heating temperature of 100 to 200 ° C. And reaction times within 2 hours are preferred. Since the particles are hydrophobic, it can be confirmed that a TMCTS film is formed on the particle surface.

(Functional group immobilization process)
Subsequent to the above pretreatment step, a functional group immobilization step is performed. A compound containing a functional group to be immobilized needs to have a double bond at its end, but there is no particular limitation other than that. For example, the structure used may have any structure between the functional group and the double bond site. In addition, the functional group is not limited to one and may be plural. A plurality of different types of functional groups may be immobilized.

  For example, in the reaction process of immobilizing a functional group such as an epoxy group or a carboxyl group, the Si-H portion contained in TMCTS and the double bond portion of the compound containing a functional group such as an epoxy group or a carboxyl group are mutually connected. The functional group is introduced into the particle surface by reaction. Specifically, the particles obtained in the pretreatment step are dispersed in a solvent, and in a heated state, the reaction catalyst and the compound having a functional group to be immobilized are added and reacted for several hours. The solvent to be used may be any type of solvent as long as it can dissolve the compound having the functional group to be immobilized and can obtain a stable reaction rate even when heated to 60 ° C. or higher. And ethylene glycol. Similarly, any kind of catalyst may be used as long as the reaction catalyst promotes the above-described reaction. For example, chloroplatinic acid or the like can be used.

  Next, an embodiment of “particles in which the polymer is adhered to the particle body” will be described below.

  In a preferred embodiment, the polymer is applied to a part of the surface of the particle body, and the “substance or functional group capable of binding the target substance” is immobilized on the surface of the particle body and / or the polymer. ing. In another embodiment, the polymer covers the entire surface of the particle body, and the “substance or functional group capable of binding the target substance” is immobilized on the surface of the polymer. When the polymer covers the entire surface of the particle body, the particles may be referred to as “encapsulated particles” or “core-shell structured particles” based on the morphology of the particles.

  The polymer deposited on the surface of the particle main body is preferably one that contributes to immobilization of the “substance capable of binding the target substance” or the “functional group to which the target substance can bind”. ”Or“ functional group to which the target substance can bind ”, the use conditions of the particles, other necessary characteristics, and the like. Examples of typical deposited polymers are polystyrene or derivatives thereof, poly (meth) acrylic acid, poly (meth) acrylic acid ester, polyvinyl ether, polyurethane, polyamide, polyvinyl acetate, polyvinyl alcohol, polyallylamine and polyethyleneimine. And at least one synthetic polymer compound selected from the group consisting of: In addition, it is not limited to such a synthetic polymer compound, These modified products or copolymers may be used. Furthermore, it may be a polymer such as a semi-synthetic polymer compound such as hydroxyalkyl cellulose, carboxyalkyl cellulose or sodium alginate, or a natural polymer compound such as chitosan, chitin, starch, gelatin or gum arabic. A polymer in which a functional group to which a “substance capable of binding a target substance” or a “functional group to which a target substance can bind” can be bonded or attached may be introduced.

  When the main purpose is to suppress the elution of metal ions (that is, metal ions constituting the particle body) from the particle surface or inside the particle, various molecules or metal ions constituting the particle body are transmitted. For example, when particles are used in an aqueous system, a polymer such as polystyrene, polyalkyl methacrylate, polyvinyl ether, or polyvinyl acetate, which is difficult to permeate water, may be selected.

  As used herein, “adhesion” substantially means an embodiment in which a polymer is attached or present on at least a part of the particle surface, and the polymer is not necessarily attached or present on the entire surface of the particle body. It does not have to be. However, in a preferred embodiment, the entire surface of the particle body is coated with the polymer so that the particle body is included in the polymer film. When the entire surface of the particle body is coated with a polymer, it is not only preferable in that the “substance or functional group capable of binding a target substance” to be immobilized on the polymer surface can be increased, but also the particle. This is preferable in that elution of metal ions (that is, metal ions constituting the particle main body) caused by the constituent material of the main body can be further suppressed.

The technique for attaching the polymer to the particle body is not particularly limited, and any technique may be used as long as the polymer can be attached to the particle body surface. For example, the following methods can be mentioned.
(1) Method of starting polymerization from the surface of the precursor particles (2) Method of polymerizing in the presence of the precursor particles and precipitating the polymer on the particle surface (3) Encapsulating the precursor particles in the monomer emulsion (4) A method in which precursor particles are mixed in a polymer solution obtained by polymerization in advance and a polymer is deposited on the particle surface.

  The above method will be described more specifically. In the method (1), an initiator or a chain transfer agent is bound or adsorbed on the surface of the precursor particle, and the polymer is extended from the particle surface. A polymer is applied to the surface. In the method (2), the polymer is deposited on the surface of the precursor particles by performing polymerization in the presence of the precursor particles using a monomer that precipitates as the polymerization reaction proceeds. The deposition can be performed more efficiently by selecting each charge so that the polymer and the particle attract each other, or fixing a polymerizable double bond on the particle surface. In the method (3), a combination of a monomer and a solvent capable of forming a monomer emulsion is selected, and the polymer particles are deposited on the particle surface by polymerizing the precursor particles in the monomer emulsion obtained thereby. Let In this case, it is preferable to use a surface treatment, a surfactant, or the like that makes the monomer particles familiar so that the precursor particles are preferentially present in the monomer emulsion. In the method (4), the precursor particles are mixed in the polymer solution, the poor solvent is added, the pH is changed, or a large amount of salt is added to lower the solubility of the polymer and precipitate. By doing so, the polymer is deposited on the surface of the precursor particles. In this case also, methods such as charge selection and fixation of polymerizable double bonds are effective. Alternatively, the precursor particles may be alternately immersed in polymer solutions having different charges to form a laminate on the particle surface.

  In the above-described method, various methods such as a microencapsulation method and emulsion polymerization are conventionally known and can be carried out using them.

  Prior to the polymer deposition treatment, the surface of the precursor particles may be subjected to a specific treatment. For example, in addition to magnetizing treatment, coating treatment with metal or inorganic substance, surfactant adsorption treatment, treatment using reactive substances such as silane coupling agent or titanium coupling agent, siloxane coating treatment, and Si- in siloxane Functional group introduction treatment (hydrosilylation reaction), acid treatment or alkali treatment, solvent washing treatment, polishing treatment or the like may be performed on H. These treatments remove the dirt on the surface of the precursor particles, control the charge on the surface of the precursor particles, and introduce reactive functional groups onto the particle surface, improving the efficiency of polymer deposition, The adhesion between the deposited polymer and the particle surface is improved. In the case where a silicon-containing substance such as siloxane or silane coupling agent is used, in addition to the “substance or functional group capable of binding to the target substance” and the deposited polymer, the silicon-containing substance is present on the surface of the particle body. It will be understood that it will be present (eg, a silicon compound may be interposed between the particle body surface and the deposited polymer surface). When polymerization is performed by previously bonding or adsorbing an initiator and / or a polymerizable double bond to the surface of the precursor particle, the surface of the deposited polymer is likely to be precipitated, which may be advantageous for polymer deposition treatment. In addition, treatments for imparting other characteristics such as reduction of non-specific binding, suppression of elution of metal ions and the like, adjustment of density, provision of color, fluorescence, and the like may be performed.

The polymer to be deposited may be subjected to a crosslinking treatment. When the deposited polymer is cross-linked, properties such as durability, solvent resistance or low swellability of the deposited polymer can be improved. The method of crosslinking is not particularly limited, but the typical methods are classified as follows.
(1) a. Crosslinking during polymer deposition treatment on precursor particles, b. Crosslinking after deposition treatment (2) a. Adding a crosslinking agent (including crosslinking that proceeds at room temperature or low temperature), b. Introducing a crosslinkable functional group into the polymer (3) a. Thermal crosslinking, b. Radiation cross-linking

  It should be noted that the above methods (1), (2) and (3) can be combined in various ways. For example, as an example of combining three of (1) a and (2) a and (3) a, polymerization is started from the surface of the precursor particle, or a polymer is deposited on the surface of the precursor particle. A method of performing a heat treatment including a bifunctional monomer in the process of depositing the polymer, or a process of performing a heat treatment including a bifunctional monomer in the process of performing polymerization by encapsulating the precursor particles in the monomer emulsion. The technique to perform can be mentioned. In addition, as an example of combining three of (1) b and (2) a and (3) a, in a system in which precursor particles are deposited by precipitation of a polymer having a carboxyl group or polymerization of a monomer having a carboxyl group A method of adding a polyfunctional epoxy cross-linking agent after application and cross-linking by applying heat can be mentioned. In the same system, a method of crosslinking using a hydroxyl group instead of a carboxyl group and an isocyanate crosslinking agent instead of an epoxy crosslinking agent is also conceivable. (2) As an example belonging to b, there is a method of introducing an epoxy group, an isocyanate group, a double bond, or the like into the deposited polymer. Here, (3) a can be used for introduction of an epoxy group or isocyanate group, and (3) b can be used for introduction of a double bond.

  When an adherent polymer is used, it is understood that "substance that can bind to the target substance" or "functional group that can bind to the target substance" is immobilized on the surface of the main body of the particle and / or the surface of the attached polymer. Like.

  The method of immobilizing the “functional group to which the target substance can bind” is not particularly limited in the embodiment in which the adhesion polymer is provided on the surface of the particle body, and the “functional group to which the target substance can bind” is not limited. Any method may be used as long as it can be bonded or attached to the substrate. Furthermore, the “functional group to which the target substance can bind” may be immobilized before, during or after the polymer deposition process.

  In the embodiment in which the adherend polymer is provided on the surface of the particle body, a method of immobilizing the “functional group to which the target substance can bind” is, for example, a “functionality to which the target substance can bind” in the polymerization reaction of the polymer to be deposited There is a method of polymerizing or copolymerizing a monomer having a “group”. In this case, examples of the monomer having a “functional group to which the target substance can bind” include (meth) acrylic acid, glycidyl (meth) acrylate, hydroxyalkyl (meth) acrylate, and dimethylaminoalkyl (meth) acrylate. And (meth) acrylic acid isocyanate alkyl, p-styrenesulfonic acid (salt), dimethylolpropanoic acid, N-alkyldiethanolamine, (aminoethylamino) ethanol or lysine.

  In the embodiment in which the adherent polymer is provided on the surface of the particle body, when it is desired to immobilize the “functional group having higher binding property to the target substance”, it is reactive to the functional group a introduced into the adherent polymer by the above method. A compound having two functional groups, that is, another functional group b having the above and “functional group c having higher binding property to the target substance” may be additionally introduced into the particles. In this case, the functional group “a” and the functional group “b” are bonded to each other, thereby obtaining particles in which “the functional group“ c ”having higher binding property to the target substance” is immobilized. In addition, when it is desired to separate the surface of the adhered polymer from the “functional group to which the target substance can bind” or to separate between the particle body surface and the “functional group to which the target substance can bind” (that is, “ Similarly, a compound having two functional groups, ie, another functional group b reactive to the introduced functional group a and a “functional group to which the target substance can be bound” May be additionally introduced into the particles having the functional group a introduced therein (in this case as well, the “functional group to which the target substance can bind” is similarly formed through the binding of the functional group a and the functional group b). Immobilized on the particles). Such a compound may be introduced twice or more times to make the linker longer. When the surface of the adherend polymer and the “functional group to which the target substance can bind” are further apart, or when the distance between the surface of the particle body and the “functional group to which the target substance can bind” is further separated, The degree of freedom of the “functional group capable of bonding to” increases not only the reactivity but also the degree of freedom of the target substance, and the advantageous effect such that the function of the target substance is not suppressed can be expected. When the number of atoms from the main chain of the adhered polymer to the functional group is defined as the length of the linker, the above effect can be expected particularly when the length of the linker is 5 to 50 atoms. Incidentally, as the linker main chain, it is particularly preferable to use a non-specific adsorptive one such as an ecological substance (for example, polyethylene glycol chain).

  The method of immobilizing the “substance that can bind to the target substance” in the embodiment in which the deposition polymer is provided on the surface of the particle body is not particularly limited, and the “substance that can bind the target substance” is bound to the particle or Any method may be used as long as it can be adhered. The “substance to which the target substance can bind” may be immobilized either before, during or after the polymer deposition process.

  For example, the “substance capable of binding to the target substance” can be immobilized on the particles by the same technique as that for introducing the “functional group to which the target substance can bind”. For example, a functional group having a binding property with a “substance that can be bound by a target substance” is previously introduced on the surface of the particle body or the surface of the adhered polymer, and the “substance that can be bound by the target substance” is transmitted via the functional group. Can be immobilized on the particles. In addition, when a hydrophobic polymer is used as the deposition polymer and a hydrophobic substance is used as the “substance that can bind to the target substance”, a so-called “hydrophobic interaction” occurs in which the hydrophobic substances are adsorbed in water. Therefore, a hydrophobic “substance capable of binding to the target substance” can be immobilized on the surface of the deposited polymer.

  Next, in the following, the binding mode between the particles and the target substance will be described. When the particle and the target substance coexist, the target substance is bound to the particle by the adsorption force or affinity acting between the “substance or functional group capable of binding the target substance” and the target substance. For the purposes of classification and explanation, “adsorption” is synonymous with “chemical adsorption”.

  As an example of the mode in which the target substance binds to the particles due to the “adsorptive power”, the “target substance” is avidin, the particle body is formed of zirconia, and “the target substance can be bound. This is the case when the “substance or functional group” is an epoxy group.

Regarding the “affinity”, the “substance or functional group capable of binding the target substance” immobilized on the surface of the particle main body is broadly classified into the following five, based on the type of affinity acting with the target substance. (Note that the substances or functional groups listed in each classification are merely examples, and other substances or functional groups are also conceivable). In addition, when it originates in such an affinity, "the substance or functional group which can bind a target substance" is called "the substance or functional group which has affinity" below.

(1) Example of “substance or functional group having affinity” in which the affinity acting with the target substance is derived from electrostatic interaction, π-π interaction, π-cation interaction or dipole interaction < Silica, activated carbon, sulfonic acid group, carboxyl group, diethylaminoethyl group, triethylaminoethyl group, phenyl group, arginine, cellulose, lysine, polylysine, polyamide, poly (N-isopropylacrylamide), crown ether or π electron (2) “Affinity substance or functional group” whose affinity acting with the target substance is due to hydrophobic interaction, such as a cyclic compound, or a functional group derivative thereof, an oxygen conjugate or a fluorescent probe conjugate examples <br/> alkyl group, an octadecyl group, an octyl group, cyanopropyl group or butyl group or a phenyl group, Others, their functional derivatives, such as oxygen conjugate or fluorescent probe conjugate (3) Example of "substance or functional group having affinity" of affinity is due to hydrogen bond acting between the target substance DNA, RNA , Oligo (dT), chitin, chitosan, amylose, cellulose, dextrin, dextran, pullulan, polysaccharide, lysine, polylysine, polyamide, poly (N-isopropylacrylamide) or β-glucan, or functional group derivatives thereof, oxygen bond (4) Examples of “substances or functional groups having affinity” in which the affinity acting between the target substance and the target substance originates from coordinate binding. Iminodiacetic acid, nickel, nickel ion, nickel Complex, cobalt, cobalt ion, cobalt complex, copper, copper ion or copper complex, or Such as oxygen conjugate or fluorescent probe conjugate al (5) Examples of "substance or functional group having affinity" of affinity is due to biochemical interaction acting between the target substance (biochemical interactions: Includes interactions related to biomolecules, including antigen-antibody reactions, ligand-receptor bonds, hydrogen bonds, coordination bonds, hydrophobic interactions, electrostatic interactions, π-π interactions, π-cation interactions, (Dipole interaction and van der Waals force)
Antigen, antibody, receptor, ligand, biotin, avidin, streptavidin neutravidin, silica, activated carbon, magnesium silicate, hydroxyapatite, albumin, amylose, cellulose, lectin, protein A, protein G, S protein, dextrin, dextran, pullulan , Polysaccharide, calmodulin, nickel, nickel ion, nickel complex, cobalt, cobalt ion, cobalt complex, copper, copper ion, copper complex, gelatin, N-acetylglucosamine, iminodiacetic acid, aminophenylboric acid, ethylenediaminediacetic acid, aminobenz Amidine, arginine, lysine, polylysine, polyamide, diethylaminoethyl group, triethylaminoethyl group, ECTEOLA-cellulose, fibronectin, vitrone Tin, arginine-glycine-asparagine (RGD) acid sequence peptide, laminin, poly (N-isopropylacrylamide), collagen, concanavalin A, adenosine 5 ′ phosphate (ATP), ADP, ATP, nicotinamide adenine dinucleotide, Acridine dye, aprotinin, ovomucoid, inhibitors such as trypsin inhibitor and protease inhibitor, phosphorylethanolamine, phenylalanine, protamine, cibacron blue, procion red, heparin, glutathione, DIG, DIG antibody, DNA, RNA, Oligo (dT), Chitin, chitosan, β-glucan, calcium phosphate, calcium hydrogen phosphate, hyaluronic acid, elastin, sericin or fibroin, or their functions Derivatives, oxygen conjugate or fluorescent probe conjugates such as

  As can be seen from the above classification, “having affinity” as used herein refers to electrostatic interaction, π-π interaction between a target substance and a substance or functional group immobilized on a particle. , Π-cation interaction, dipole interaction, hydrophobic interaction, biochemical interaction, hydrogen bond or coordination bond, and the like are substantially meant. Note that depending on the type of substance or functional group immobilized on the particle body, two or more of the above-mentioned affinity may be combined, and there may be a substance or functional group overlapping in the above classification. . Moreover, it is not necessarily limited to the above-mentioned classification, and any substance or functional group can be attached to the particle as long as it has a function of acting on the target substance and causing the target substance to exist on or near the particle surface. They may be immobilized (for example, those having affinity due to a complementary shape with the target substance are considered).

  The mode or method for immobilizing the “substance or functional group having affinity for the target substance” on the surface of the particle serving as the precursor is not limited at all. For example, by interposing a polymer, an inorganic compound, a low molecular linker or a coupling agent, etc., a “substance or functional group having affinity for the target substance” can be bound, adsorbed or adsorbed on the surface of the unfixed particle. It can be immobilized by absorption. Further, the “substance or functional group having affinity for the target substance” can also be immobilized on the surface of the unfixed particle by using a general particle coating method.

  An example of a technique for immobilizing “a substance having affinity for a target substance” on the particle body will be described. For example, when an antibody is used as a “substance having affinity for a target substance”, a double bond and an epoxy group are bonded to the Si—H portion of the siloxane by the above-described functional group immobilization method using siloxane. A compound having glycidyl methacrylate is reacted to fix an epoxy group on the particle surface. Further, the antibody can be immobilized on the particle by stirring the obtained particle with the antibody in water.

  In addition, a method for immobilizing “functional group having affinity for a target substance” on the particle surface can be performed by a method using siloxane as described above, for example.

  Next, the microdevice of the present invention will be described in detail below. The microdevice of the present invention includes the above-described particles and “a member provided with a plurality of depressions”, and the plurality of depressions are configured to accommodate the particles.

  The “member provided with a plurality of depressions” is preferably a member in which a plurality of depressions are provided in alignment on at least one surface of the member, such as a well plate member or a bio or chemical chip. is there. The well plate member includes a microwell plate, a microplate, a deep well plate, a shallow well plate, a multiwell plate, a microtiter plate, and the like. In addition, for bio or chemical chips, bio chips, chemical chips, reactor chips, DNA chips, protein chips, cell chips, sensing chips, pretreatment chips that are generally used for measurement or inspection in the bio field or medical field, etc. , A μTAS chip or a biochemical fluid system.

  When the “member provided with a plurality of depressions” is a well plate member, the depression substantially refers to a “well”. A necessary number of such depressions may be provided according to the application (or use). For example, about 10 to 10 million depressions are provided in alignment with the member. There is no particular limitation on the shape of the opening surface of the dent (that is, “the shape when the dent is viewed from above the dent” or “the cross-sectional shape when the dent is cut off in a plane parallel to the surface of the member”). For example, as shown in FIG. 1, a circle (FIG. 1A), an ellipse (FIG. 1B), a diamond (FIG. 1C), a rectangle (FIG. 1D), a rectangle (FIG. 1 (e)) or a polygon such as a regular hexagon (FIG. 1 (f)). Further, the cross-sectional shape in the depth direction of the depression is not particularly limited, and as shown in FIG. 2, a quadrangle (FIG. 2 (a)), a rectangle (FIG. 2 (b)), a triangle (FIG. 2 (c)), It may be trapezoid (FIG. 2 (d)), circular (FIG. 2 (e)), oval (FIG. 2 (f)), or the like. Furthermore, the edge part of an opening part or a bottom face part may be formed in the curve (refer FIG.2 (g)).

  When supplying the liquid in which the particles are dispersed to the “member provided with a plurality of depressions” so that the individual positions of the particles are determined, a plurality of particles may be put into one depression. In many cases, it is desirable that one particle enters one depression, for example, when different substances are bonded to each bead or different treatments are performed (see FIG. 3). Therefore, when it is assumed that one particle is accommodated in one depression with the opening surface of the depression horizontal, the opening surface of the depression is higher than the center of the particle or the same height (or equivalent). In the case of (height), it is preferable that the diameter D1 of the circle inscribed with respect to the cross-sectional shape of the recess cut by the horizontal plane passing through the center of the particle is 100% to 190% of the average particle size d (hereinafter, “ Referred to as “Case A”). In other words, in case A, it is preferable that D1 = 1.0d to 1.9d. Alternatively, in the same assumption, when the opening surface of the depression is lower than the center of the particle, the diameter D2 of the circle inscribed in the opening shape of the depression is preferably 10 to 190% of the average particle size d. (Hereafter referred to as “Case B”). In other words, in case B, it is preferable that D2 = 0.1d to 1.9d.

  If D2 is smaller than 10% of the average particle size d, it will be difficult to stably hold the particles in the recess, whereas if D1 and D2 are larger than 190% of the average particle size d, there will be 2 particles in one recess. Since it becomes easy to line up individual pieces, it becomes difficult to specify the positions of individual particles. That is, when D1 is 100% to 190% of the average size d and D2 is 10% to 190% of the average size d of the particles, the liquid in which the particles are dispersed is “a member provided with a plurality of depressions”. When the particles are supplied, one particle is accommodated in each of the depressions. D1 is more preferably 100 to 175% of the average particle size d, more preferably 100 to 150% of the average particle size d, and D2 is 35 to 35% of the average particle size d. It is more preferably 175%, and further preferably 50 to 150% of the average particle size d.

Here, the “case A” and “case B” will be described in more detail as follows. First, with respect to case A, “when the opening surface of the dent is horizontal and one particle is accommodated in one dent” is an embodiment as shown in FIG. “PL1” in the cross-sectional view of FIG. 4A corresponds to the opening surface of the recess provided horizontally. Next, regarding the case A, “the cross-sectional shape of the dent cut out on the horizontal plane passing through the center of the particle” means the cross-sectional shape of the dent cut out on the surface of “PL2” shown in FIG. 4B (S represented by the dotted line). _A ) means. Then, for the case A a "circle inscribed against depression sectional shape", as shown in FIG. 4 (c), which means that the circle C _A represented by solid lines, according circle C _A The diameter corresponds to D1. That is, “case A” has D1 of 100 to 190% of the average particle size d. Next, “Case B” will be described. Regarding the case B, “when the opening surface of the dent is lower than the center of the particle” means that when the particle is accommodated in the dent, the opening surface PL1 of the dent is lower than the center CEp of the particle as shown in FIG. Substantially means that it exists below. And about the case B, "the shape of the opening part of a hollow" means "S _B " represented by the dotted line in FIG.5 (b). Hence, for case B the "circle inscribed against recess of the opening shape" means that the circle C _B represented by the solid line shown in FIG. 5 (c), the diameter of such circle Corresponds to D2. That is, “Case B” has D2 of 10 to 190% of the average particle size d. Incidentally, when D2 is 100% to 190% of the average size d of the particles, the particles and the depressions may have an embodiment as shown in FIG.

  When the inclination of the side wall surface inside the dent is less than 90 ° with respect to the opening surface of the dent, the force acts in the direction in which the particle exits the dent when the particle collides with the inner wall or is pressed. It can be said that the particles once accommodated tend to come out of the depression. In the case of particles having a high density used in the microdevice of the present invention, since gravity acts strongly, the particles are prevented from jumping out of the dent compared to particles having a low density. In other words, it can be said that the effect of the present invention is more exhibited when the inclination of the side wall surface inside the depression is smaller than 90 ° with respect to the opening surface of the depression. If the angle is small, not only the particles are likely to enter the recess, but also the “sample containing the target substance” and the like easily flows into the recess, which is advantageous in various applications.

  Here, when the inclination of the side wall surface inside the depression is 5 ° to 88 ° with respect to the opening surface of the depression, the effect of the present invention is particularly high and preferable. If the inclination of the side wall surface inside the dent is smaller than 5 °, it is difficult to stably hold the particles in the dent, and if the inclination of the side wall surface inside the dent is larger than 88 °, the particles are stably held in the dent even if the particle has a low density. This is because the advantage is relatively small although the advantage is possible. The inclination of the side wall surface inside the recess is more preferably 10 ° to 85 °, and further preferably 25 ° to 80 °. As used herein, “inclination of the side wall surface of the recess” or “inclination of the side wall surface of the recess” means that the cross section of the side wall surface inside the recess is linear as shown in FIG. In this case, the angle α1 formed by the opening surface PL1 of the dent and the side wall surface inside the dent is substantially meant, and when the cross section of the side wall surface inside the dent is curved as shown in FIG. As shown in FIG. 6 (c), the angle α2 that is the maximum among the angles formed by the tangent line TL of the curve U and the opening surface PL1 of the depression substantially means.

  Similarly to the above, if the depth of the dent is shallow, a force acts in a direction in which the particle exits the dent when the particle collides with the inner wall or is pressed, so that it can be said that the particles once accommodated easily exit the dent. In the case of particles having a high density used in the microdevice of the present invention, since gravity acts strongly, the particles are prevented from jumping out of the dent compared to particles having a low density. In other words, in the microdevice of the present invention, the depth of the recess may be relatively shallow, such as 5% to 200% of the average particle size d, for example, 10% to 150% of the average particle size d. . When the depth of the dent is shallow as described above, the “sample containing the target substance” or the like easily flows to the inside of the dent, so that advantageous effects can be obtained in various applications. As shown in FIG. 2, the “depth of the depression” here substantially refers to the distance Ld from the opening surface of the depression to the deepest bottom of the depression.

  The pitch Pt (or interval Pt) of the openings of “plurality of depressions” as shown in FIG. 7 is preferably 0 to 3 times the particle size d, more preferably 0 to 2 times the particle size d. It is. Accordingly, when the liquid in which the particles are dispersed is supplied to the “member provided with a plurality of depressions”, it is promoted that one particle is accommodated in each of the depressions. In particular, the microdevice of the present invention may have an aspect in which the pitch of the plurality of recesses is substantially zero as shown in FIGS. 8A and 8B (that is, the plurality of recesses). The opening surface of each other may be continuous without being separated from each other). This is because the particles used in the microdevice of the present invention are particles having a relatively high density, and when the liquid in which the particles are dispersed is supplied to the “member provided with a plurality of depressions”, it settles faster due to natural sedimentation. This is because it may be preferable that the interval between the plurality of depressions is as narrow as possible.

  The material of the micro device may be any material as long as there is no problem in the application. As a typical example of the material of the microdevice, at least one material selected from the group consisting of silicon, polydimethylsiloxane, polycarbonate, polyolefin, polycyclic olefin, polyacrylate ester and polystyrene can be given.

  Any method may be used as a method of providing a plurality of depressions in the microdevice. Typical examples include dry etching, wet etching, photolithography, (nano) imprint, injection molding, cutting, and the like. In the (nano) imprint system, hot emboss molding (2T method), UV curable resin molding (2P method), or the like can be suitably used.

  In a preferred embodiment, the device of the present invention is pre-stored with a plurality of depressions. In this specification, “particles are contained in a plurality of depressions” substantially means an aspect in which particles are contained in at least a part of the plurality of depressions, and particles are contained in all the depressions. It does not have to be. Preferably, one particle is previously stored in each of the plurality of depressions. In order to accommodate the particles in the plurality of depressions in advance, for example, it is preferable to supply a liquid (for example, water) in which the particles are dispersed to the “member provided with the plurality of depressions”. More specifically, the particles are preferably accommodated in the plurality of depressions by the method described in step (i) of the treatment method of the present invention described later. Incidentally, the liquid solvent component may be evaporated by allowing the particles to be accommodated in the dent and then subjecting to standing or drying.

  In another preferred embodiment, the “member provided with a plurality of recesses” may be a member having at least two surfaces provided with the recesses. In this case, it is preferable that “at least two or more surfaces provided with depressions” are provided in a multistage state and are in communication with each other via a flow path.

  Next, the “processing method for analyzing, extracting, purifying or reacting a target substance using a microdevice” will be described below.

  The processing method of the present invention includes (i) a step of preparing a “member provided with a plurality of depressions”, and storing one particle in each of the plurality of depressions of the member, and (ii) The method substantially includes a step of supplying a sample containing a target substance to the particles contained in the depression and bringing the particles into contact with the sample.

  In step (i), as shown in FIG. 9, by supplying “a liquid in which particles 12 are dispersed” to “a member provided with a plurality of depressions”, one particle is provided for each of the plurality of depressions. The particles are accommodated one by one (note that it is sufficient that the particles are accommodated in at least one part of the plurality of depressions, and the particles may not necessarily be accommodated in all the depressions). As described above, the “member provided with a plurality of depressions” is, for example, a well plate member or a bio or chemical chip. The “liquid in which the particles are dispersed” is preferably particles on at least one medium selected from the group consisting of water, alcohols, organic solvents such as DMF and DMSO, and buffers mainly composed of these. It may be prepared by dispersing. The content of particles in the liquid is preferably 0.1 to 50% by volume.

  When the “liquid in which particles are dispersed” is supplied to the “member provided with a plurality of depressions”, it is preferably performed in a manner as shown in FIG. That is, the lid member 20 is provided for the “member 10 provided with a plurality of depressions”, and the “liquid 30 in which particles are dispersed” is reciprocated in a certain direction or reciprocated so as to pass between them. It is preferable to repeat the process, or to apply a tilt, vibration or motion after the supply, or to apply a magnetic field if the magnet has magnetism. The lid member 20 may be provided with projections at positions corresponding to the plurality of depressions.

  In the treatment method of the present invention, when the “liquid in which particles are dispersed” is supplied to the “member provided with a plurality of depressions” due to the relatively high density of the particles, the particles contained in the liquid Will be housed in the depression by natural sedimentation. Also, due to the relatively high density of the particles, once entering the dent, the particles are less likely to jump out of the dent. Accordingly, a degree of freedom is provided in the flow of the liquid 30 when the “liquid 30 in which particles are dispersed” is provided to the “member 10 provided with a plurality of depressions” (for example, the liquid 30 is supplied at a relatively high speed). Even if it flows, the particles will be contained in the depressions). As described above, since the size of the depression is preferably 10 to 190% of the average particle size d of D1 or D2, one particle is accommodated for each of the depressions. It will be.

  In the step (ii), the sample and the particles are brought into contact with each other by supplying the “sample including the target substance” to the “member provided with a plurality of depressions”. Specifically, the sample 40 passes between the member 10 and the lid member 20 so that contact between the “particles 12 provided in the plurality of depressions” and the “sample 40 including the target substance” is brought about. In this way, the liquid is flowed in a certain direction (see FIG. 10B). Note that the target substance contained in the sample is bound to the particles when the sample and the particles come into contact with each other.

  In the treatment method of the present invention, due to the relatively high density of the particles, even if the sample is flowed in step (ii), the particles are unlikely to jump out of the depression and tend to be held at a certain position. is doing. That is, it can be said that the degree of freedom of the flow of the sample 40 when the “sample 40 including the target substance” is provided to the “member 10 provided with a plurality of depressions” is large (for example, the sample 40 is moved at a relatively high speed). Even if flowed, the particles remain in the recesses). Further, in the processing method of the present invention, the inclination of the side wall surface inside the recess can be reduced due to the relatively high density of particles (specifically, the angle shown in FIG. 6 (a)). α1 or the angle α2 shown in FIG. 6 (c) can be reduced), or the depth of the depression can be made shallower, so that in step (ii), the sample can be easily moved into the depression or As a result, the binding between the particles and the target substance in the sample is further promoted.

  After the step (ii) is performed, particles on which the target substance is immobilized can be obtained. By using or applying the particles, various kinds of cells, proteins, nucleic acids, chemical substances, and the like can be obtained. The target substance can be analyzed, extracted, purified, reacted, and the like. More specifically, for example, in the “method for analyzing a target substance”, the amount of the target substance is absorbed and chemiluminescent using, as a marker, an enzyme, a fluorescent dye, a magnetic substance, or the like that binds to the target substance. Quantitative analysis or qualitative analysis of a target substance by detecting it by fluorescence, magnetism, etc., or by causing a luminescence or color reaction using a reaction or by-product at the time of binding to the target substance. it can. In addition, in the “method for extracting a target substance” or “method for purifying a target substance”, after collecting the particles, by using “a substance that releases and releases the target substance from the particles” or necessary heating, cooling, etc. The target substance can be extracted or purified by performing the above-described process. Furthermore, in the “method of reacting a target substance”, the reaction of the target substance can be performed by performing mixing, heating, stirring, ultraviolet irradiation, etc. in the depression of the “member provided with a plurality of depressions”.

  Although the embodiments of the present invention have been described above, the present invention is not limited thereto, and those skilled in the art will readily understand that various modifications can be made.

  In order to confirm “natural particle sedimentation rate”, “particle binding characteristics”, “stable retention of particles against dents” and the like, the following examples and comparative examples were carried out.

<Preparation of particles>
In Examples 1-20 and Comparative Examples 1-8, particles were prepared as follows.

Example 1
The particles prepared in Example 1 are yttrium-added zirconia particles P ₁ having a hydroxyl group immobilized thereon. First, it was prepared a yttrium-doped zirconia particles p ₁ of Nikkato Corporation. The particles p ₁ had a particle size of 50 μm, a specific surface area of 0.02 m ² / g, and a density of 6 g / cm ³ . 1g of particles _{p 1} were dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particle p ₁ was heated in a thermostatic bath at 150 ° C. for 1.5 hours. By such processing, change as particle p ₁ has a hydrophobic, it was confirmed that particles p ₁ is coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Next, the particles were subjected to washing, filtration, and drying to obtain yttrium-added zirconia particles P ₁ having hydroxyl groups immobilized on the surface. The particles P ₁ was hydrophilic. The specific surface area of the particle P ₁ was 0.02 m ² / g, the density was 6 g / cm ³ , and the particle size was about 50 μm (the difference between the particle size of the particle P _{1 and} the particle size of p ₁ was measured). Within the error range).

(Example 2)
Particles prepared in Example 2 is a yttrium-doped zirconia particles P ₂ where the hydroxyl group is immobilized. The particle P ₂ is different from the particle P _{1 of} Example 1 in that the particle P ₂ has magnetism.

First, it was prepared a yttrium-doped zirconia particles p ₂ of Nikkato Corporation. The particles p ₂ had a particle size of 50 μm, a specific surface area of 0.02 m ² / g, and a density of 6 g / cm ³ . 1g of particles p ₂ were dispersed in water, the silane coupling agent to the resultant dispersion (available from Shin-Etsu Chemical Co., KBM-903) by the addition of a silane coupling agent to the surface of the particle p ₂ to be I wore it. Then the plating nucleus is generated in Shipley Far ist made Pd catalyst Catalyst-6F added the surface of the particle _{p 2.} After washing the obtained particles with 1.2N hydrochloric acid, a magnetic nickel plating layer is formed on the particle surface using nickel plating solution Top Nicolon LPH manufactured by Okuno Pharmaceutical Co., Ltd., and the particles are washed, filtered and dried. It was attached. Thereafter it may implement the same process as in Example 1 to obtain particles P ₂ where the hydroxyl group is immobilized. That is, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried, hydroxyl groups on the surface to obtain an immobilized yttrium-doped zirconia particles P _2. The particles P ₂ was hydrophilic. The specific surface area of the particle P ₂ was 0.02 m ² / g, the density was 6 g / cm ³ , and the particle size was about 50 μm (the difference between the particle size of the particle P _{2 and} the particle size of the particle p ₂ is Within measurement error range). The saturation magnetization of the particle P ₂ was measured and found to be 4.5 A · m ² / kg.

(Example 3)
Particles prepared in Example 3 is a yttrium-doped zirconia particles P ₃ which hydroxyl group is immobilized. The method for preparing the particles is different from that in Example 1.

First, it was prepared a yttrium-doped zirconia particles p ₃ manufactured by NIKKATO CORPORATION. The particles p ₃ had a particle size of 50 μm, a specific surface area of 0.02 m ² / g, and a density of 6 g / cm ³ . 10 g of particles p3 were dispersed in 25 g of pure water, and ₃ g of 3-glycidoxypropyltrimethoxysilane having an epoxy group at the end was added to the resulting dispersion and stirred for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried, hydroxyl group to obtain a yttrium-doped zirconia particles P ₃ which is immobilized on the surface. The particles P ₃ were hydrophilic. The specific surface area of the particle P ₃ was 0.02 m ² / g, the density was 6 g / cm ³ , and the particle size was about 50 μm (the difference between the particle size of the particle P _{3 and} the particle size of the particle p ₃ is Within measurement error range).

Example 4
Particles prepared in Example 4 is a yttrium-doped zirconia particles P ₄ which hydroxyl group is immobilized. The method for preparing the particles is different from that in Examples 1 and 3.

First, it was prepared a yttrium-doped zirconia particles p ₄ manufactured by NIKKATO CORPORATION. The particle p ₄ had a particle size of 50 μm, a specific surface area of 0.02 m ² / g, and a density of 6 g / cm ³ . 10g of the particle p ₄ were dispersed in pure water 25 g, and stirred for 4 hours with the addition of tetraethoxysilane 5g and aqueous ammonia 5g respect resulting dispersion. Thereafter, 3 g of 3-glycidoxypropyltrimethoxysilane having an epoxy group at the terminal was added to the dispersion and stirred for 3 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Thereafter, the particles were subjected to washing, filtration, and drying to obtain yttrium-added zirconia particles P ₄ having a hydroxyl group immobilized on the surface. The particles P ₄ were hydrophilic. The specific surface area of the particle P ₄ was 0.02 m ² / g, the density was 6 g / cm ³ , and the particle size was about 50 μm (the difference between the particle size of the particle P _{4 and} the particle size of the particle p ₄ is Within measurement error range).

(Example 5)
Particles prepared in Example 5 is a yttrium-doped zirconia particles P ₅ which avidin is immobilized.

First, it was prepared a yttrium-doped zirconia particles p ₅ manufactured by NIKKATO CORPORATION. Such particles _{p 5} had a particle size of 50 [mu] m, a specific surface area of 0.02 m ² / g, a density of 6 g / cm ^3. The particles p ₅ of 10g were dispersed in pure water 25 g, while stirring the resulting dispersion was stirred for an additional 4 hours with the addition of 3-glycidoxypropyltrimethoxysilane 3g having an epoxy group at the terminal to the dispersion . Next, the particles were washed with acetone, and then subjected to vacuum drying to obtain yttrium-doped zirconia particles having an epoxy group.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was applied to 200 mg of the obtained particles, and stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, avidin was obtained yttrium-doped zirconia particles _{P 5} immobilized. The obtained particle P _{5 had} a specific surface area of 0.02 m ² / g, a density of 6 g / cm ³ , and a particle size of about 50 μm (the particle size of the particle P _{5 and} the particle size of the particle p ₅ were The difference is within the measurement error range).

(Example 6)
Particles prepared in Example 6 are alumina particles P ₆ which avidin is immobilized. The raw material particles and the preparation method are different from those in Example 5.

First of all, we were prepared alumina particles p ₆ made of Taimei Chemicals. Such particles _{p 6} had a particle size of 200 [mu] m, a specific surface area of 0.008 m ² / g, a density of 3.6 g / cm ^3. 1g of particles _{p 6} were dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particle _{p 6} was heated for 1.5 hours in a thermostatic bath at 0.99 ° C.. By such processing, changes as particles p ₆ has a hydrophobic, it was confirmed that particles p ₆ was coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

  Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, the dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Subsequently, the particles were subjected to washing, filtration, and drying to obtain alumina particles having hydroxyl groups immobilized on the surface. Such particles were hydrophilic.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, to obtain alumina particles _{P 6} which avidin is immobilized. The specific surface area of the obtained particle P ₆ was 0.008 m ² / g, the density was 3.6 g / cm ³ , and the particle size was about 200 μm (the particle size of the particle P _{6 and} the particle size of the particle p ₆ The difference from the diameter is within the measurement error range).

(Example 7)
Particles prepared in Example 7, copper particles P ₇ which avidin is immobilized. The raw material particles are different from Example 6.

First of all, we were prepared copper particles p ₇ made of Hitachi Metals. Such particles _{p 7} had a particle size of 50 [mu] m, a specific surface area of 0.013 m ² / g, a density of 8.9 g / cm ^3. The 1g of particles _{p 7} are dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particle _{p 7} was heated for 1.5 hours in a thermostatic bath at 0.99 ° C.. By such processing, changes as particles p ₇ has a hydrophobicity, it was confirmed that particles p ₇ was coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

  Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, the dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Subsequently, the particles were subjected to washing, filtration, and drying to obtain copper particles having hydroxyl groups immobilized on the surface. Such particles were hydrophilic.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying to obtain copper particles _{P 7} which avidin is immobilized. The obtained particle P _{7 had} a specific surface area of 0.013 m ² / g, a density of 8.9 g / cm ³ , and a particle size of about 50 μm (the particle size of the particle P _{7 and} the particle size of the particle p ₇ The difference from the diameter is within the measurement error range).

(Example 8)
Particles prepared in Example 8 is a yttrium-doped zirconia particles P ₈ which avidin has been immobilized. The raw material particles are different from those of Example 6 and Example 7. Example 8 is the same as Example 5 in that the raw material particles are made of yttrium-added zirconia, but Example 8 uses yttrium-added zirconia having physical properties different from those of Example 5.

First, it was prepared a yttrium-doped zirconia particles p ₈ manufactured by NIKKATO CORPORATION. Such particles _{p 8} had a particle size of 30 [mu] m, a specific surface area of 0.03 m ² / g, a density of 6 g / cm ^3. The 1g of particles _{p 8} were dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particles _{p 8} was heated for 1.5 hours in a thermostatic bath at 0.99 ° C.. By such processing, the particles p ₈ is changed to hydrophobic, it was confirmed that particles p ₈ is covered with 1,3,5,7-tetramethylcyclotetrasiloxane.

  Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, the dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Next, the particles were subjected to washing, filtration, and drying to obtain yttrium-added zirconia particles having hydroxyl groups immobilized on the surface. Such particles were hydrophilic.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, to obtain a yttrium-doped zirconia particles _{P 8} which avidin has been immobilized. The obtained particle P _{8 had} a specific surface area of 0.03 m ² / g, a density of 6 g / cm ³ , and a particle size of about 30 μm (the particle P ₈ particle size and the particle p ₈ particle size The difference is within the measurement error range).

Example 9
Particles prepared in Example 9 is a yttrium-doped zirconia particles P ₉ with avidin immobilized. Example 9 is the same as Example 5 and 8 in that the raw material particles are made of yttrium-doped zirconia, but Example 9 uses yttrium-doped zirconia having physical properties different from those of Examples 5 and 8.

First, it was prepared a yttrium-doped zirconia particles p ₉ manufactured in thermal Corporation. Such particles _{p 9} had a particle size of 15 [mu] m, a specific surface area of 0.04 m ² / g, a density of 6 g / cm ^3. The 1g of particles _{p 9} dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particles _{p 9} was heated for 1.5 hours in a thermostatic bath at 0.99 ° C.. By such processing, the particles p ₉ is changed to hydrophobic, it was confirmed that particles p ₉ is covered with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried, hydroxyl group to obtain a yttrium-doped zirconia particles P ₉ immobilized on the surface. Such particles were hydrophilic.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, to obtain a yttrium-doped zirconia particles _{P 9} with avidin immobilized. The obtained particle P _{9 had} a specific surface area of 0.04 m ² / g, a density of 6 g / cm ³ , and a particle size of about 15 μm (the particle size of the particle P _{9 and} the particle size of the particle p ₉ The difference is within the measurement error range).

(Example 10)
Particles prepared in Example 10, an epoxy group is a polystyrene deposited zirconia particles P ₁₀ immobilized. Such particles are characterized in that a polymer is deposited on at least a part of the particle body.

First, it was prepared a yttrium-doped zirconia particles p ₁₀ manufactured by NIKKATO CORPORATION. Such zirconia particles _{p 10} had a particle size of 30 [mu] m, a specific surface area of 0.03 m ² / g, a density of 6 g / cm ^3. The zirconia particles p ₁₀ of 3g was dispersed in water / alcohol mixed solution, and stirred for about 30 minutes methacrylonitrile added trimethoxysilane 0.13 g 35 ° C.. Then, nitrogen bubbling was performed for 30 minutes, 0.1 g of sodium p-styrenesulfonate, 0.1 g of potassium persulfate, 9.4 g of styrene monomer, and 1.4 g of glycidyl methacrylate were added and reacted at 70 ° C. for 8 hours. After completion of the reaction, the slow particles unreacted material and precipitation by removal by washing with water, an epoxy group was obtained immobilized polystyrene deposited zirconia particles P _10. The obtained particle P _{10 had} a specific surface area of 0.05 m ² / g, a density of 5.5 g / cm ³ , and a particle size of about 30 μm (the particle size of the particle P ₁₀ and the particle p ₁₀ The difference from the diameter is within the measurement error range). The specific surface area of the particles P ₁₀ obtained was although more than the specific surface area of the raw material particles p _10, which is that portion of the polymer was deposited on the particulate considered as one of the causes.

(Example 11)
Particles prepared in Example 11, avidin is a polystyrene deposited zirconia particles P ₁₁ immobilized. Point avidin in place of the epoxy groups have been immobilized is different from the particles P ₁₀ Example 10.

200 mg of the “polystyrene-coated zirconia particles P ₁₀ with an epoxy group immobilized” obtained in Example 10 was applied to an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of a 10 mM PBS solution (pH 7.2), and stirred overnight. did. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, avidin obtain polystyrene deposition zirconia particles _{P 11} immobilized. The obtained particle P _{11 had} a specific surface area of 0.05 m ² / g, a density of 5.5 g / cm ³ , and a particle size of about 30 μm (the particle size of the particle P ₁₁ and the particle p ₁₀ The difference from the diameter is within the measurement error range). Although the specific surface area of the obtained particles P ₁₁ is increasing than the specific surface area of the raw material particles p _10, which, as in Example 10, that portion of the polymer was deposited on the particulate Cause 1 It can be considered as one.

Example 12
Particles prepared in Example 12, an epoxy group is crosslinked polystyrene deposited zirconia particles P ₁₂ immobilized. That the deposition polymer polystyrene is crosslinked is different from the particles P ₁₀ Example 10.

By performing the same treatment as in Example 10 except that 0.3 g of divinylbenzene was used as a crosslinking agent for the styrene monomer of Example 10, “crosslinked polystyrene-coated zirconia particles P _{12 with} an epoxy group immobilized” were obtained. Obtained. The obtained particle P _{12 had} a specific surface area of 0.05 m ² / g, a density of 5.5 g / cm ³ , and a particle size of about 30 μm (the particle size of the particle P _{12 and} the particle of the particle p ₁₀ The difference from the diameter is within the measurement error range). Although the specific surface area of the obtained particles P ₁₂ is increasing than the specific surface area of the raw material particles p _10, which, as in Example 10, that portion of the polymer was deposited on the particulate Cause 1 It can be considered as one.

(Example 13)
Particles prepared in Example 13 is a magnetic zirconia particles P ₁₃ polystyrene depositions epoxy group is immobilized. The particle P ₁₃ is different from the particle P _{10 of} Example 10 in that the particle P ₁₃ has magnetism.

First, it was prepared a yttrium-doped zirconia particles p ₁₃ manufactured by NIKKATO CORPORATION. Such particles _{p 13} had a particle size of 30 [mu] m, a specific surface area of 0.03 m ² / g, a density of 6 g / cm ^3. The 1g of particles p ₁₃ were dispersed in water, a silane coupling agent to the resultant dispersion (available from Shin-Etsu Chemical Co., KBM-903) by adding, to the surface of the particle p ₁₃ a silane coupling agent to be I wore it. Was then generated surface in the plating nuclei particles _{p 13} was added to Shipley Far ist made Pd catalyst Catalyst-6F. After the obtained particles were washed with 1.2N hydrochloric acid, a nickel plating layer was formed on the particle surface using a nickel plating solution top Nicolon LPH manufactured by Okuno Pharmaceutical, and the particles were subjected to washing, filtration and drying. . After this, by carrying out the same treatment as in Example 10, "epoxy group to obtain a magnetic zirconia particles P ₁₃ immobilized polystyrene deposited. The specific surface area of the particles P ₁₃ is 0.05m ² / g, the density was 6.5 g / cm ³ , the particle size was 32 μm, and the saturation magnetization of the particle P ₁₃ was measured to be 6.5 A · m ² / kg. Incidentally, the specific surface area of the obtained particle P ₁₃ is larger than the specific surface area of the raw material particle p ₁₃ , and this is because 1 part of the polymer was deposited in the same manner as in Example 10. It can be considered as one.

(Example 14)
Particles prepared in Example 14 is a yttrium-doped zirconia particles P ₁₄ that "anti-human CRP monoclonal antibody 6404" was immobilized. First, it was prepared a yttrium-doped zirconia particles p ₁₄ manufactured by NIKKATO CORPORATION. Such particles _{p 14} had a particle size of 50 [mu] m, a specific surface area of 0.02 m ² / g, a density of 6 g / cm ^3. The 1g of particles _{p 14} were dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particle _{p 14} was heated for 1.5 hours in a thermostatic bath at 0.99 ° C.. By such processing, changes as particles p ₁₄ has a hydrophobic, it was confirmed that particles p ₁₄ is covered with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, the particles were washed with water, and then the dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Subsequently, the particles were subjected to washing, filtration, and drying to obtain yttrium-added zirconia particles P14 _′ having a hydroxyl group immobilized on the surface. Such particles P14 _′ had hydrophilicity.

Tosyl chloride was added to the particles P _{14 ′} and stirred. The obtained particles were washed to obtain tosyl group activated zirconia particles. Anti-human CRP monoclonal antibody 6404 (manufactured by MedixBiochemica) was immobilized on the tosyl group-activated zirconia particles. In addition, it confirmed that the anti-human CRP monoclonal antibody 6404 was fix | immobilized on the particle | grain surface from the color development by a HRP-Rabbit-Anti-Mouse IgG2a secondary antibody (made by ZYMED). The specific surface area of the particles _{P 14} obtained by the above operations, a 0.02 m ² / g, density of 6 g / cm ^3, a particle size of 50 [mu] m.

(Example 15)
Particles prepared in Example 15 is a yttrium-doped zirconia particles P ₁₅ which hydroxyl group is immobilized. The method for preparing the particles is different from that in Example 1.

First, it was prepared a yttrium-doped zirconia particles p ₁₅ manufactured by NIKKATO CORPORATION. Such particles _{p 15} had a particle size of 50 [mu] m, a specific surface area of 0.02 m ² / g, a density of 6 g / cm ^3. The particles p ₁₅ of 1g dispersed in water, relative to the resulting dispersion, was added dropwise with mixed KBE-402 (manufactured by Shin-Etsu Chemical Co.) in ethanol. Aqueous ammonia was added thereto, and the mixture was stirred at room temperature for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried to obtain yttrium-doped zirconia particles P ₁₅ which hydroxyl group is immobilized on the surface. The particles _{P 15} was hydrophilic. The specific surface area of the particle P ₁₅ was 0.02 m ² / g, the density was 6 g / cm ³ , and the particle size was about 50 μm (the difference between the particle size of the particle P _{15 and} the particle size of the particle p ₁₅ is Within measurement error range).

(Example 16)
The particles prepared in Example 16 are yttrium-doped zirconia particles P ₁₆ in which avidin is immobilized on the particles of Example 1.

An aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was applied to 200 mg of the particles obtained in Example 1, and stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, to obtain a yttrium-doped zirconia particles _{P 16} with avidin immobilized. The specific surface area of the obtained particles _{P 16} is a 0.02 m ² / g, density of 6 g / cm ^3, a particle size of about 50 [mu] m.

(Example 17)
Particles prepared in Example 17 is tungsten particles P ₁₇ with avidin immobilized.

First, it was prepared Hitachi Metals made of tungsten particles p _17. Such particles _{p 17} had a particle size of 100 [mu] m, a specific surface area of 0.003 m ² / g, a density of 19.1 g / cm ^3. The 1g of particles _{p 17} were dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particles were heated in a constant temperature bath at 150 ° C. for 1.5 hours. By such processing, it changes as the particle p ₁₇ with hydrophobic, it was confirmed that particles coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried, hydroxyl groups on the surface to obtain a tungsten particles p ₁₇ immobilized. Such particles p ₁₇ had hydrophilicity.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, to obtain a tungsten particles _{P 17} with avidin immobilized. The specific surface area of the particles P ₁₇ was 0.003 m ² / g, the density was 19.1 g / cm ³ , and the particle size was about 100 μm.

(Example 18)
Particles prepared in Example 18 is a lead particles P ₁₈ with avidin immobilized. Only the raw material particles are different from Example 17.

First, it was prepared Ohashi steel balls made of lead particles p _18. Such particles _{p 18} had a particle size of 800 [mu] m, a specific surface area of 0.00066m ² / g, a density of 11.36 g / cm ^3. The 1g of particles _{p 18} were dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particles were heated in a constant temperature bath at 150 ° C. for 1.5 hours. By such processing, changes as particles p ₁₈ has a hydrophobic, it was confirmed that particles coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried to obtain a lead particles p ₁₈ a hydroxyl group on the surface is fixed. Such particles p ₁₈ had hydrophilicity.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, to obtain a lead particles _{P 18} with avidin immobilized. The specific surface area of the particles P ₁₈ was 0.00066 m ² / g, the density was 11.3 g / cm ³ , and the particle size was about 800 μm.

Example 19
Particles prepared in Example 19, a tungsten particles P ₁₉ with avidin immobilized. The particle adjustment method is different from Example 17.

First of all, we were prepared tungsten particles p ₁₉ manufactured by Hitachi Metals. The particle p ₁₉ had a particle size of 100 μm, a specific surface area of 0.003 m ² / g, and a density of 19.1 g / cm ³ . The 10g of the particle p ₁₉ were dispersed in pure water 25 g, while stirring the resulting dispersion was stirred for an additional 4 hours with the addition of 3-glycidoxypropyltrimethoxysilane 3g having an epoxy group at the terminal to the dispersion . Then, after washing the particles with acetone, by subjecting the particles to vacuum drying, to obtain a tungsten particles p ₁₉ having an epoxy group.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was applied to 200 mg of the obtained particles, and stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, to obtain a tungsten particles _{P 19} with avidin immobilized. The specific surface area of the particles P ₁₉ was 0.003 m ² / g, the density was 19.1 g / cm ³ , and the particle size was about 100 μm.

(Example 20)
Particles prepared in Example 20 is tungsten particles P ₂₀ with avidin immobilized. Particles _{P 20} is different from the particles _{P 17} Example 17 in that magnetic.

First of all, we were prepared tungsten particles p ₂₀ manufactured by Hitachi Metals. Such particles _{p 20} had a particle size of 100 [mu] m, a specific surface area of 0.003 m ² / g, a density of 19.1 g / cm ^3. The 1g of particles p ₂₀ were dispersed in water, a silane coupling agent to the resultant dispersion (available from Shin-Etsu Chemical Co., KBM-903) by adding, to the surface of the particle p ₂₀ a silane coupling agent to be I wore it. Next, a Pd catalyst Catalyst-6F manufactured by Shipleyfarist was added to generate plating nuclei on the surface of the particles. After the obtained particles are washed with 1.2N hydrochloric acid, a nickel plating layer is formed on the particle surface using the nickel plating solution Top Nicolon LPH manufactured by Okuno Pharmaceutical, and the particles are subjected to washing, filtration and drying. did. After this, by carrying out the same treatment as in Example 17 to obtain particles p ₂₀ which hydroxyl group is immobilized. That is, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried to obtain tungsten particles p ₂₀ which hydroxyl group is immobilized on the surface. Such particles _{p 20} were hydrophilic.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was applied to 200 mg of the obtained particles, and stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, avidin was obtained immobilized tungsten particles _{P 20.} The specific surface area of the particles _{P 20} is a 0.003 m ² / g, density 19.1 g / cm ^3, a particle size of about 100 [mu] m. Incidentally, was 13A ^· m 2 / kg was measured saturation magnetization of the particles _{P 20.}

  Incidentally, in the above-mentioned examples, an example using a silane coupling agent having an epoxy group is included. As the silane coupling agent, a compound having a mercapto group, a functional group having a double bond, etc. Note that may be used.

(Comparative Example 1)
Particles prepared in Comparative Example 1 is a cross-linked acrylic particles hydroxyl group is immobilized R _1.

First, it was prepared a cross-linked acrylic particles r ₁ manufactured by Soken Chemical & Engineering Co., Ltd.. The particles r ₁ had a particle size of 30 μm, a specific surface area of 0.033 m ² / g, and a density of 1.19 g / cm ³ . 1 g of particles r1 were dispersed in toluene, and ₁ g of 1,3,5,7-tetramethylcyclotetrasiloxane (manufactured by Shin-Etsu Chemical Co., Ltd., LS-8600) was added to the resulting dispersion. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. The particles were then heated for 1.5 hours in a constant temperature bath at 150 ° C. By this treatment, it was confirmed that the particles r ₁ changed to hydrophobic and the particles were coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 20 mg of chloroplatinic acid and 1 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, the particles were washed with water, and then the dispersion obtained by supplying 20 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried, hydroxyl groups on the surface to obtain an immobilized crosslinked acrylic particles R _1. Such particles R ₁ had hydrophilicity. The specific surface area of the particle R ₁ was 0.033 m ² / g, the density was 1.19 g / cm ³ , and the particle size was about 30 μm (the particle size of the particle R _{1 and} the particle size of the particle r ₁ The difference is within the measurement error range).

(Comparative Example 2)
The particles prepared in Comparative Example 2 are porous zeolite particles R ₂ on which avidin is immobilized.

First, zeolite particles (HSZ-700) r ₂ manufactured by Tosoh Corporation were prepared. The particles r ₂ had a particle size of 18 μm, a specific surface area of 170 m ² / g, and a density of 2.3 g / cm ³ . 1 g of particles r2 was dispersed in toluene, and ₂ g of 1,3,5,7-tetramethylcyclotetrasiloxane (manufactured by Shin-Etsu Chemical Co., Ltd., LS-8600) was added to the resulting dispersion. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. The particles were then heated for 1.5 hours in a constant temperature bath at 150 ° C. By such processing, change as the particle r ₂ have hydrophobic, it was confirmed that particles coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

The obtained particles were dispersed in water and heated to 80 ° C. 20 mg of chloroplatinic acid and 2 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. And after washing | cleaning particle | grains with water, the dispersion liquid obtained by providing 20 ml of 10 wt% ethanolamine with respect to particle | grains was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried, hydroxyl groups on the surface to obtain an immobilized porous zeolite particles r _2. Such particles r ₂ was hydrophilic.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. Thereafter, the particles were washed with a 10 mM PBS solution (pH 7.2) and water, and then subjected to vacuum drying to obtain porous zeolite particles R ₂ having avidin immobilized thereon. The obtained particle R _{2 had} a specific surface area of 170 m ² / g, a density of 2.3 g / cm ³ , and a particle size of about 18 μm (the particle size of the particle R _{2 and} the particle size of the particle r ₂ The difference is within the measurement error range).

(Comparative Example 3)
Particles prepared in Comparative Example 3 are silica particles R ₃ with avidin immobilized. The raw material particles are different from those of Comparative Example 2.

First of all, were prepared silica particles r ₃ made NIPPN techno cluster. The particles r ₃ had a particle size of 3.0 μm, a specific surface area of 1.2 m ² / g, and a density of 1.96 g / cm ³ . The particles _{r 3} of 1g dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particles were heated in a constant temperature bath at 150 ° C. for 1.5 hours. By such treatment, the silica particles r ₃ is changed to the hydrophobic silica particles r ₃ was confirmed to have been coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Next, the particles were subjected to washing, filtration, and drying to obtain silica particles silica particles r ₃ having hydroxyl groups immobilized on the surface. Such particles r ₃ had hydrophilicity.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying to obtain silica particles _{R 3} with avidin immobilized. The resulting specific surface area of the particles _{R 3} a 1.2 m ² / g, density was 1.96 g / cm ^3, a particle size of about 3.0 [mu] m (particle size and particle _{r 3} particles _{R 3} The difference from the particle size is within the measurement error range).

(Comparative Example 4)
Particles prepared in Comparative Example 4, avidin is zirconia particles R ₄ with immobilized porous structure. The raw material particles are different from those of Comparative Examples 2 and 3.

First, it was prepared ZirChrom Co. porous zirconia particles (ZirChrom-PHASE) r _4. The particle r ₄ had a particle size of 25 μm, a specific surface area of 30 m ² / g, and a density of 6 g / cm ³ . 1g of particles _{r 4} are dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particles were heated in a constant temperature bath at 150 ° C. for 1.5 hours. By such processing, the particles r ₄ is changed to hydrophobic, we were confirmed that particles r ₄ is covered with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried, hydroxyl group to obtain a porous zirconia particles r ₄ immobilized on the surface. Such particles r ₄ were hydrophilic.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, avidin obtain zirconia particles R ₄ with immobilized porous structure. The obtained particle R _{4 had} a specific surface area of 30 m ² / g, a density of 6 g / cm ³ , and a particle size of about 25 μm (difference between the particle size of the particle R _{4 and} the particle size of the particle r ₄ Is within the measurement error range).

(Comparative Example 5)
Particles prepared in Comparative Example 5 is a porous silica particles R ₅ in which avidin has been immobilized. The raw material particles are different from those of Comparative Examples 2 to 4.

First, Asahi Glass Si-Tech made of porous silica particles (Sunsphere L-121) were prepared r _5. The particle r ₅ had a particle size of 11.5 μm, a specific surface area of 336 m ² / g, and a density of 2.0 g / cm ³ . The particles _{r 5} of 1g dispersed in toluene, 1,3,5,7-tetramethylcyclotetrasiloxane against dispersion obtained (manufactured by Shin-Etsu Chemical Co., LS-8600) was added 0.5 g. After the dispersion was evaporated to evaporate toluene, the particles were allowed to stand at 50 ° C. for 4 hours in a vacuum desiccator. Thereafter, the particles were heated in a constant temperature bath at 150 ° C. for 1.5 hours. By such processing, the particle r ₅ varies so as to have hydrophobicity, it was confirmed that the particles r ₅ coated with 1,3,5,7-tetramethylcyclotetrasiloxane.

Subsequently, the obtained particles were dispersed in water and heated to 80 ° C. 10 mg of chloroplatinic acid and 0.5 g of Kyoeisha Chemical Light Ester were added to the resulting dispersion and stirred at 80 ° C. for 4 hours. Next, after the particles were washed with water, a dispersion obtained by supplying 10 ml of 10 wt% ethanolamine to the particles was stirred at room temperature for 12 hours. Then, by subjecting the particles to washed, filtered and dried to obtain a porous silica particles r ₅ which hydroxyl group is immobilized on the surface. Such particles r ₅ had hydrophilicity.

Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying, avidin obtain a porous silica particles _{R 5} immobilized. The obtained particle R _{5 had} a specific surface area of 336 m ² / g, a density of 2.0 g / cm ³ , and a particle size of about 11.5 μm (the particle size of the particle R _{5 and} the particle size of the particle r ₅ The difference from the diameter is within the measurement error range).

(Comparative Example 6)
Particles prepared in Comparative Example 6, the epoxy group is a crosslinked acrylic particles R ₆ immobilized. This is different from Comparative Example 1 in terms of the functional group to be immobilized.

First, it was prepared a cross-linked acrylic particles r ₆ manufactured by Soken Chemical & Engineering Co., Ltd.. The particle r ₆ had a particle size of 30 μm, a specific surface area of 0.03 m ² / g, and a density of 1.19 g / cm ³ . The particles r ₆ of 10g were dispersed in pure water 25 g, relative to the resulting dispersion was added 3-glycidoxypropyltrimethoxysilane 3g having an epoxy group at the end was stirred for 4 hours. Then, by subjecting the particles to washed, filtered and dried, epoxy group to obtain a crosslinked acrylic particles R ₆ immobilized on the surface. The obtained particle R _{6 had} a specific surface area of 0.03 m ² / g, a density of 1.19 / cm ³ , and a particle size of about 30 μm (the particle size of the particle R _{6 and} the particle size of the particle r ₆ The difference from the diameter is within the measurement error range). Note that such particles R ₆ was hydrophilic.

(Comparative Example 7)
The particles prepared in Comparative Example 7 are porous zeolite particles R ₇ on which avidin is immobilized. The manufacturing method is different from that of Comparative Example 2.

First, Tosoh Co., Ltd. of the zeolite particles (HSZ-700) were prepared _{r 7.} The particle r ₇ had a particle size of 18 μm, a specific surface area of 170 m ² / g, and a density of 2.3 g / cm ³ . The particles r ₇ of 10g was dispersed in pure water 25 g, relative to the resulting dispersion was added 3-glycidoxypropyltrimethoxysilane 3g having an epoxy group at the end was stirred for 4 hours. Then, by subjecting the particles to washed, filtered and dried, epoxy group to obtain a porous zeolite particles r ₇ immobilized on the surface. Such particles r ₇ had hydrophilicity. Subsequently, an aqueous solution in which 100 mg of avidin was dissolved in 20 ml of 10 mM PBS solution (pH 7.2) was added to 200 mg of the obtained particles, and the mixture was stirred overnight. After washing the particles with 10mMPBS solution (pH 7.2) and water, by subjecting the particles to vacuum drying to obtain zirconia particles _{R 7} in which avidin has been immobilized. The obtained particle R _{7 had} a specific surface area of 170 m ² / g, a density of 2.3 g / cm ³ and a particle size of about 18 μm (the particle size of the particle R _{7 and} the particle size of the particle r ₇ The difference is within the measurement error range).

(Comparative Example 8)
Particles _{R 8} prepared in Comparative Example 8 is Dynabeads M-280 Tosylactivated particles "anti-human CRP monoclonal antibody 6404" was immobilized.

Such particles _{R 8} are added and stirred Dynabeads M-280 Tosylactivated particles (Dynal Ltd.) anti-human CRP monoclonal antibody 6404 (manufactured by MedixBiochemica), was washed with magnetic separation, anti-human CRP monoclonal antibody 6404 (produced by MedixBiochemica) It was prepared by immobilizing on the particle surface. In addition, it was confirmed that the anti-human CRP monoclonal antibody 6404 was immobilized on the particle surface from color development by the HRP-Rabbit-Anti-Mouse IgG2a secondary antibody. The specific surface area of the obtained particle R ₈ was 6 m ² / g, the density was 1.3 g / cm ³ , and the particle size was 2.8 μm.

Various conditions of the above Examples 1-20 and Comparative Examples 1-8 are shown in Table 1.

<Confirmation test of particle natural sedimentation rate>
In order to confirm the natural sedimentation rate of the particles obtained in the examples and comparative examples, the following tests were conducted.

  First, 1 g of the particles obtained in Examples and Comparative Examples was dispersed in 5 ml of water in a test tube and allowed to stand. Then, the time (hereinafter also referred to as “natural sedimentation time”) until the transparent supernatant was obtained after standing was measured. The natural sedimentation rate can be indirectly determined from the natural sedimentation time. Moreover, the same operation was performed in the state which has arrange | positioned the magnet near the bottom part of the test tube. The results are shown in Table 2.

From the results in Table 2, the following matters can be grasped.

(A) The natural sedimentation rate of the particles of the high density example is generally much faster than the natural sedimentation rate of the particles of the comparative example. In other words, when a liquid in which particles having a relatively high density as dispersed in the microdevice of the present invention are dispersed is supplied to a “member provided with a plurality of depressions”, a relatively high density composed of resin, silica, or the like. Compared to the lower particles, they enter the pit more quickly due to spontaneous sedimentation.

(B) Comparing natural sedimentation rates between Comparative Examples 2, 3, 5 and 7 having the same density (1.96 to 2.3 g / cm ³ ), the naturalness of Comparative Example 5 having a large specific surface area Sedimentation speed is slowest. Although it is a porous particle of the raw material particle of Comparative Example 5, such a porous particle having a large specific surface area (that is, a particle having an internal penetrating network structure) has many voids and gas exists inside the particle. It is estimated that it became difficult to settle. That is, it can be said that the natural sedimentation rate is slow for porous particles having a large specific surface area. Although the specific surface area of the particles used in the microdevice of the present invention is relatively small, the particles enter the recess more rapidly from the viewpoint of such a small specific surface area.

(C) Further, when the comparison between Example 1 and Example 2, or the comparison between Example 10 and Example 13 or the comparison between Example 17 and Example 20, is performed, the particles having magnetism are obtained. The particles can be moved faster by supplementing the magnetic separation operation in addition to the natural sedimentation.

<< Production of "members with multiple indentations">>
A “member provided with a plurality of depressions” was produced as follows.

(Prototype pattern 1)
A circular mask was placed on a metal material and wet etching was performed to obtain a plate-like member having a large number of depressions having a shape of a part of a spherical surface. (See FIG. 11 (a)).

(Prototype pattern 2)
A polycarbonate plate was drilled with a 0.07 mm cutter to create a cylindrical recess. As a result, a plate-like member having a large number of columnar depressions was obtained (see FIG. 11B).

(Prototype pattern 3)
Grooves parallel to three directions at 120 degrees were formed in the metal material by cutting to form a number of regular hexagonal protrusions. This was formed into a shape, and transferred by an imprint method using a UV curable resin (2P method) to obtain a plate-like member having many regular hexagonal columnar depressions (see FIG. 11C).

<Confirmation test for stable retention of particles against dent>
In order to confirm the penetration into the dent and the stable retention of the particles obtained in the examples and comparative examples, the following tests were performed.

  First, as shown in FIG. 12, on the obtained plate-like member 10, a silicone rubber member 15 having a hole 15c opened at a position corresponding to a depression is placed as a packing, and a glass plate 20 is placed as a lid member, and a flow path is obtained. Formed. In addition, notches (15a, 15b) are provided in two opposing directions of the silicone rubber member 15, and the liquid can flow between the plate-like member 10 and the glass plate 20 in a certain direction. A flow path is formed. Water was allowed to flow through this flow path, and the particles obtained in the examples and comparative examples were poured together with water. At this time, the ease of entering and exiting the particles was observed by changing the flow of water, tilting in various directions, and applying vibration to the plate-like member 10. The particles used are the particles used in Examples 1, 6 and 7, and the particles used in Comparative Examples 1, 4 and 5.

The results are shown in Table 4 below. In Table 4, as an index of ease of entry of particles into the dent, “◎” indicates easy entry, “○” indicates that particles are generally reciprocated many times, and still some do not enter “△” indicates a thing and “×” indicates a thing that hardly enters (shown on the left side in the grid of Table 4). On the other hand, as an index of the ease of getting out of the dent of particles, “◎” is the one that hardly appears even when the flow velocity is increased or vibration is given, and the one that goes out little by little when the flow velocity is increased or vibration is given. “◯”, “Δ” means that it is relatively easy to go out by the same operation, and “X” means that it is almost gone by a slight flow or vibration (shown on the right side of the square in Table 4).

  From the results of Table 4, in the microdevice of the present invention, when the liquid in which the particles are dispersed is supplied to the “member provided with a plurality of depressions”, the particles easily enter the depressions, and the flow and vibration of the liquid of the sample and the like It turned out that it was hard to jump out. Accordingly, it will be understood that the degree of freedom in flowing the sample and the like becomes high and handling becomes easy.

<< Confirmation test of surface condition of raw material particles >>
The surface state of the raw material particles used in Example 1 and Comparative Example 5 was observed using a Hitachi scanning electron microscope (SEM, model S-4500). The results are shown in FIGS. 13 and 14 are electron micrographs of the yttrium-doped zirconia particles p ₁ used in Example 1, and FIGS. 15 and 16 are electron micrographs of the porous silica particles r ₅ used in Comparative Example 5. As apparent from FIGS. 13 to 16, the particles of Example 1 are non-porous particles, and the particle surface is smooth without irregularities, whereas the particles of Comparative Example 5 are porous particles, The surface irregularities are large and bumpy. From this, it can be understood that the specific surface area is significantly different, that is, the specific surface area of the particles used in the microdevice of the present invention is smaller.

  17 (a) and 17 (b) are sectional views of the vicinity of the surface of the yttrium-doped zirconia particles, which are the raw material particles used in the examples, and FIGS. 18 (a) and 18 (b) are used in Comparative Example 4. Sectional drawing of the surface vicinity of the raw material particle | grains (namely, porous zirconia particle) which showed was shown. As can be seen particularly from these drawings, the porous zirconia particles used in the comparative example have through-holes formed therein (the “black portions that undulate” in FIGS. 18A and 18B). The raw material particles of the examples are non-porous and have no through-holes (refer to FIGS. 17A and 17B). In other words, in the particles of the prior art finally obtained in the comparative example, the particle body has a through-hole, whereas in the “particles used in the microdevice of the present invention” finally obtained in the examples, the particles It will be particularly understood that the main body is non-porous and has no through-holes (in the “particles used in the microdevice of the present invention”, the through-holes are formed in the particle main body. This can also be understood by referring to FIGS. 13 and 14).

<Confirmation test for specific / non-specific binding properties of particles>
Particles P ₅ , Particles P ₁₁ , Particles P ₁₆ and Particles P ₁₇ obtained in Examples 5, ₁₁ , ₁₆ and ₁₇ and Particles R ₂ , Particles R ₄ obtained in Comparative Examples 2, 4, 5 and 7 using a particle _{R 5} and particles _{R 7,} confirming the specificity-nonspecific binding properties of the particles _{(P 5, P 11, P} 16, P 17, R 2, R 4, R 5, R 7). As target substances, two kinds of substances, HRP and biotinylated HRP, were used (both enzyme activities are substantially equivalent). Avidin immobilized on the particles specifically binds to biotinylated HRP, but does not specifically bind to HRP. That is, biotinylated HRP binds specifically (preferentially) to particles, while HRP can be adsorbed to the pore region of the particles and binds non-specifically to the particles.

Since the same operation was performed on each of the particles P ₅ and the particles P ₁₁ , P ₁₆ , P ₁₇ and the particles R ₂ , the particles R ₄ , the particles R ₅ , and the particles R ₇ , the particles of Example 5 are described below. It will be mainly described operation for the P _5. First, 1.5ml tube 2 was prepared and charged particles _{P 5} an appropriate amount of each (amount such coloring level is 0.01 to 1.5). 100 μl of biotinylated HRP with a concentration of 20 ng / ml was added to one tube, and 100 μl of HRP with a concentration of 20 ng / ml was added to the other tube, followed by stirring with a vortex mixer for 30 minutes. Then, of 10 mM PBS buffer (pH 7.2) at 400 [mu] l, it was subjected to wash the particles _{P 5} was charged to each tube to centrifugation. This washing and centrifugation were performed 4 times. After removal of the PBS buffer (pH 7.2), by standing by adding 200μl of TMB (tetramethylbenzidine) to each tube containing the particles _{P 5} 30 minutes, and developed the particles _{P 5.} Subsequently, 200 μl of 1N sulfuric acid was added to stop the reaction. Then, the absorbance (450 nm) was measured with a TECAN plate reader Infinite 200 to determine the color development amount of the particles P ₅ charged in each tube. The amount of color developed to particle P ₅ of coloring level and HRP was subjected to particle P ₅ biotinylated HRP is subjected, respectively, to the amount of biotinylated HRP and particles P ₅ that specifically binds to the particles P ₅ It is proportional to the amount of HRP that binds non-specifically. Therefore, when the ratio of the color development amount I of the biotinylated HRP that specifically binds to the nonspecifically bound HRP and the color development amount I nonspecific (I specific / I nonspecific) is large, the nonspecific binding characteristics of the particles Less and conversely, when such a ratio (I-specific / I non-specific) is small, the non-specific binding properties of the particles will be greater.

The same procedure, Example 11 of particles _{P 11,} particles _{P 16} Example 16, particles _{P 17} Example 17, Comparative Example grains _{R 2} 2, particles _{R 4} in Comparative Example 4, the particles of Comparative Example 5 R _5, performed even for particles R ₇ of Comparative example 7, the coloring level I nonspecific of HRP to coloring level I-specific and non-specific binding of the biotinylated HRP that binds specifically to each of the particles Asked.

The results are shown in Table 5. As can be seen from the results in Table 5, Example particles _{P 5} 5 Example particles _{P 11} of 11, the particles _{R 2} it is of Comparative Example 2 of the particles _{P 17} particle _{P 16} and Example 17 Example 16 It was found that the value of I-specific / I-nonspecific was larger than that of the particle R ₄ of Comparative Example ₄ , the particle R ₅ of Comparative Example ₅ and the particle R ₇ of Comparative Example 7, and the non-specific binding was suppressed. That is, it can be understood that non-specific binding with a substance other than the target substance can be suppressed with particles having a small specific surface area and substantially non-porous.

<Summary>
From the above “confirmation test of natural sedimentation rate of particles” and “confirmation test of non-specific binding properties of particles”, the particles used in the microdevice of the present invention have a large natural sedimentation rate, and a liquid in which particles are dispersed is expressed as “ Non-specific in which substances other than the target substance can bind to the particles when a sample containing the target substance is flowed, while being able to enter the depression more quickly when it is supplied to `` members with multiple depressions '' It was found that the binding can be suppressed. In addition, in the “confirmation test of the stability of particles against the depressions”, the microdevice of the present invention allows the particles to easily enter the depressions when the liquid in which the particles are dispersed is supplied to the “member provided with a plurality of depressions”. In addition, it was found that the particles once entering the dent are not easily ejected from the dent due to the flow of liquid or vibration of the sample or the like. Furthermore, from the “confirmation test of the surface state of the raw material particles”, it is also confirmed that the particles used in the microdevice of the present invention are non-porous particles and “the particle body does not have through holes”. I was able to.

  The microdevice of the present invention can be used for quantification, separation, purification and analysis of target substances such as cells, proteins, nucleic acids or chemical substances. For example, the microdevice of the present invention can bind nucleic acids such as DNA, and as a result can be used for DNA analysis, and thus contributes to tailor-made medical technology.

FIG. 1 is a diagram schematically showing the shape of the opening of the recess. FIG. 2 is a diagram showing a cross-sectional shape of a dent in which the shape of the dent in the depth direction can be understood. FIG. 3 is a perspective view schematically showing an aspect in which one particle is accommodated in one depression. FIG. 4 is a diagram schematically showing an aspect of case A. FIG. 5 is a diagram schematically showing an aspect of case B. FIG. 6 is a view schematically showing “inclination of the side wall surface of the depression”. FIG. 7 is a diagram schematically showing the pitch Pt of the openings of the “plurality of depressions”. FIG. 8 is a diagram schematically showing an aspect of the microdevice in which the pitch of the plurality of recesses is substantially zero. FIG. 9 is a diagram schematically showing an aspect in which “a liquid in which particles are dispersed” is supplied to “a member provided with a plurality of depressions”. FIG. 10 is a diagram schematically showing a preferred embodiment of the method of the present invention. FIG. 11 is a diagram schematically showing an aspect of the produced plate-like member. FIG. 12 is a diagram schematically showing a development view of the device used in the “confirmation test of the stability of particles against the depression”. FIG. 13 is a photograph of the non-porous yttrium-doped zirconia particles p ₁ that are the raw material particles of Example 1. FIG. 14 is an enlarged photograph of the surface of the non-porous yttrium-doped zirconia particles p ₁ that are the raw material particles of Example 1. FIG. 15 is a photograph of porous silica particles r ₅ that are the raw material particles of Comparative Example 5. FIG. 16 is an enlarged photograph of the surface of porous silica particles r ₅ that are the raw material particles of Comparative Example 5. FIG. 17A is a particle cross-sectional view in the vicinity of the surface of yttrium-doped zirconia particles, which are raw material particles used in the examples. FIG. 17B is an enlarged view of the particle surface in FIG. FIG. 18A is a particle cross-sectional view in the vicinity of the surface of porous zirconia particles that are raw material particles used in Comparative Example 4. FIG. FIG.18 (b) is the figure which expanded the particle | grain surface in Fig.18 (a) more.

Explanation of symbols

  10 ... a member provided with a plurality of depressions, 12 ... particles, 13 ... depressions, 15 ... a silicone rubber member, 15a, 15b ... notches provided in the silicone rubber member, 15c ... a silicone rubber member A provided hole, 20 ... a lid member (for example, a glass plate), 30 ... a liquid in which particles are dispersed, 40 ... a sample containing a target substance.

Claims (21)

A microdevice comprising particles capable of binding a target substance and a member provided with a plurality of depressions,
Substance or functional group capable of the surface of the particle body linking the target substance of the particles is immobilized, the density of the particles is ^{3.5g / cm 3 ~23g / cm 3} , also the plurality A microdevice characterized in that the depression can accommodate the particles.   The microdevice according to claim 1, wherein the particle body does not have a through hole. Wherein the specific surface area of the particles is 0.0005m ² / g~10m ² / g, a micro device according to claim 1 or 2. A polymer is deposited on a part of the surface of the particle body,
The microdevice according to claim 1, wherein a substance or a functional group capable of binding the target substance is immobilized on the surface of the particle body or the polymer. The polymer covers the entire surface of the particle body;
5. The microdevice according to claim 4, wherein a substance or a functional group capable of binding the target substance is immobilized on the surface of the polymer.   The polymer is at least one selected from the group consisting of polystyrene, poly (meth) acrylic acid, poly (meth) acrylic ester, polyvinyl ether, polyurethane, polyamide, polyvinyl acetate, polyvinyl alcohol, polyallylamine, and polyethyleneimine. The microdevice according to claim 4, wherein the microdevice is the above polymer.   The microdevice according to claim 4, wherein the polymer is crosslinked.   The micro device according to claim 1, wherein the particles are non-porous.   9. The particle body according to claim 1, wherein the particle body is formed of at least one material selected from the group consisting of zirconia, yttrium-added zirconia, iron oxide, and alumina. Micro device.   The micro device according to claim 1, wherein the particles have magnetism. The microdevice according to claim 10, wherein the saturation magnetization is 0.5 to 85 A · m ² / kg.   The micro device according to claim 1, wherein an average size of the particles is 1 μm to 1 mm.   The substance capable of binding the target substance is at least one substance selected from the group consisting of biotin, avidin, streptavidin, and neutravidin. A microdevice according to any of the above.   The functional group capable of binding the target substance is carboxyl group, hydroxyl group, epoxy group, tosyl group, succinimide group, maleimide group, thiol group, thioether group, disulfide group, aldehyde group, azide group, hydrazide group, primary At least one selected from the group consisting of an amino group, a secondary amino group, a tertiary amino group, an imide ester group, a carbodiimide group, an isocyanate group, an iodoacetyl group, a halogen substituent of a carboxyl group, and a double bond The micro device according to claim 1, wherein the micro device is a functional group of   The microdevice according to any one of claims 4 to 7, wherein a silicon-containing substance and / or polyethylene glycol is present on at least a part of the surface of the particle body and / or the surface of the polymer.   16. The target substance can be bound to the particle by an adsorption force or affinity acting between the target substance and a substance or functional group capable of binding the target substance. A microdevice according to any of the above.   The affinity acting between the target substance and a substance or functional group capable of binding the target substance is electrostatic interaction, π-π interaction, π-cation interaction, dipole interaction, hydrophobic The microdevice according to claim 16, characterized by being caused by an interaction, a hydrogen bond, a coordination bond or a biochemical interaction. When one particle is accommodated in one depression with the opening surface of the depression horizontal,
When the opening surface of the depression is higher than or equal to the center of the particle, the diameter D1 of the circle inscribed with respect to the sectional shape of the depression cut in a horizontal plane passing through the center of the particle is the average of the particle 100 to 190% of the size d, or
When the opening surface of the depression is lower than the center of the particle, the diameter D2 of the circle inscribed in the shape of the opening of the depression is 10 to 190% of the average size of the particle. The microdevice according to claim 1.   The micro device according to claim 1, wherein an inclination of a side wall surface of the depression is 5 ° to 88 ° with respect to an opening surface of the depression.   The micro device according to claim 1, wherein the particles are previously accommodated in the plurality of depressions. A method for performing analysis, extraction, purification or reaction of a target substance using the microdevice according to claim 1,
(I) supplying the liquid containing the particles to the member, and storing the particles one by one in each of the plurality of depressions; and (ii) on the surface of the member provided with the plurality of depressions. Supplying a sample containing a target substance to the sample and bringing the sample into contact with the particles,
In the step (i), the particles contained in the liquid are naturally settled and accommodated in the depression.