WO2009136623A1 - Sample reactor and sample reaction method - Google Patents

Sample reactor and sample reaction method Download PDF

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Publication number
WO2009136623A1
WO2009136623A1 PCT/JP2009/058640 JP2009058640W WO2009136623A1 WO 2009136623 A1 WO2009136623 A1 WO 2009136623A1 JP 2009058640 W JP2009058640 W JP 2009058640W WO 2009136623 A1 WO2009136623 A1 WO 2009136623A1
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WIPO (PCT)
Prior art keywords
sample
reaction
transfer film
adsorption
tank
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PCT/JP2009/058640
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French (fr)
Japanese (ja)
Inventor
淳典 平塚
英樹 木下
菜央 坂口
憲二 横山
祐二 丸尾
宇一 緑川
豊 鵜沼
Original Assignee
独立行政法人産業技術総合研究所
シャープ株式会社
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Publication of WO2009136623A1 publication Critical patent/WO2009136623A1/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44756—Apparatus specially adapted therefor
    • G01N27/44773—Multi-stage electrophoresis, e.g. two-dimensional electrophoresis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/405—Concentrating samples by adsorption or absorption
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/00527—Sheets
    • B01J2219/00538—Sheets in the shape of cylinders
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00596—Solid-phase processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/0068—Means for controlling the apparatus of the process
    • B01J2219/00702—Processes involving means for analysing and characterising the products
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00722—Nucleotides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00725—Peptides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/0074—Biological products

Definitions

  • the present invention relates to an apparatus and method used for reacting a sample with a reaction solution.
  • proteome is intended to mean a whole protein produced by translation in a specific cell, organ or organ, and its research includes protein profiling.
  • proteins have unique properties of charge and molecular weight, they combine a proteome, which is a mixture of many proteins, rather than separating individual proteins into their components based on charge alone or molecular weight alone. As a result, more proteins can be separated with high resolution.
  • Proteins are separated by charge and / or molecular weight by electrophoresis, but it is difficult to specify biological properties from the separation position of proteins separated only by such physical properties.
  • function of a protein is controlled by being subjected to chemical modification (post-translational modification) such as phosphorylation after being synthesized. It is difficult to obtain information on such post-translational modifications by electrophoresis alone.
  • Western blotting is a method of transferring proteins in a slab gel separated by electrophoresis to a membrane. If a specific antibody is overlaid on the membrane onto which the protein has been transferred by Western blotting, the protein can be specified to some extent based on the antigen-antibody reaction.
  • phosphorylation which is one of post-translational modifications, it is possible to detect the presence or absence of phosphorylation and the difference in phosphorylation sites by overlaying an anti-phosphorylated protein antibody on the membrane onto which the protein has been transferred. .
  • Overlaying a specific antibody on the membrane onto which the protein has been transferred usually involves reacting the transfer membrane with each of blocking buffer, primary antibody, washing solution, and secondary antibody in a plastic container or plastic bag, etc. Is done by letting These operations are usually performed manually. At this time, in order to promote the reaction, the plastic container or the plastic bag is reciprocated or rotated. A detection method as described in Patent Document 1 has also been developed.
  • JP 2004-163146 A publication date: June 10, 2004
  • the present invention has been made in view of the above problems, and a main object of the present invention is to provide a sample reaction apparatus that uniformly reacts a sample transferred to a transfer film and a reaction solution.
  • a sample reaction apparatus stores an adsorption structure having a sample adsorption region for adsorbing a sample, a reaction solution, and the sample adsorption region is in contact with the reaction solution.
  • a reaction tank that houses the adsorption structure; and a drive unit that rotates the adsorption structure in the reaction tank.
  • the sample adsorption region is provided on a surface that covers the rotation axis of the rotation. It is characterized by being.
  • the sample adsorption region rotates in a continuous manner like a water wheel in the reaction solution. Therefore, compared to the case where the sample is reciprocated as in the prior art, the sample and the reaction solution can be reacted uniformly, and compared with the case where the entire reaction vessel is rotated as in the conventional technique, Since unevenness of the reaction solution accompanying the movement of the rotating tank can be avoided, there is an effect that the sample and the reaction solution can be reacted uniformly.
  • the surface covering the rotation axis is preferably a surface parallel to the rotation axis.
  • the sample adsorption region is provided on a plane parallel to the rotation axis. Therefore, the sample adsorption region is composed of a line orthogonal to the rotation about the rotation axis. Therefore, the samples on each line have the same moving speed with the rotation, and the sample and the reaction liquid can be uniformly contacted.
  • the surface covering the rotation axis is preferably a cylindrical surface or an elliptical cylindrical surface.
  • the adsorption structure may further include a transfer film provided with the sample adsorption region and a transfer film support that supports the transfer film.
  • a gap is provided between the transfer film support and the transfer film. Further, it is preferable that the transfer film support is provided with a groove along a direction obliquely intersecting with a straight line parallel to a rotation axis of rotation by the driving unit.
  • the reaction solution can pass through the back side of the transfer film. Therefore, the flow of the reaction liquid becomes easy, and there is an effect that the sample and the reaction liquid can be reacted more uniformly.
  • the adsorption structure may further include a cover that covers the sample adsorption region.
  • the cover allows the reaction solution to pass through.
  • transmits the said reaction liquid
  • suction structure is accommodated in the said reaction tank with this cover attached, and by rotating, the said sample, the said reaction liquid, There is an effect that can be reacted.
  • the inner surface of the reaction vessel may be a semi-cylindrical surface shape or a wall surface shape when a polygonal column equal to or higher than a hexagonal column is cut by a plane passing through a central axis parallel to the length direction. preferable.
  • the said reaction tank has a shape close
  • a groove or a protrusion along a direction obliquely intersecting with a straight line parallel to the rotation axis may be provided on the inner surface of the reaction vessel.
  • the flow of the reaction liquid generated by the rotation of the adsorption structure along the direction of the rotation is slanted by the grooves or protrusions along the direction that obliquely intersects the straight line parallel to the rotation axis. Since the reaction solution is flowed not only in the direction of rotation but also in an oblique direction and the reaction solution is further stirred, the sample and the reaction solution are reacted more uniformly. There is an effect that can be.
  • the reaction tank may further include an adding means for adding the reaction liquid and a waste liquid tank for receiving the reaction liquid flowing out from the reaction tank.
  • the sample may be a protein
  • the reaction solution may contain an antibody of the protein
  • the sample reaction apparatus may include a plurality of the reaction vessels, and the driving unit may move the adsorption structure to any reaction vessel.
  • the sample is protein
  • the plurality of reaction tanks are a first reaction tank that stores a blocking buffer, a second reaction tank that stores a reaction liquid containing a primary antibody that binds to the protein, and the primary reaction tank.
  • the sample reaction apparatus also includes a transfer film support that supports a transfer film that adsorbs a sample and a reaction liquid, and the transfer film held by the transfer film support is in contact with the reaction liquid.
  • the transfer film support covers the rotation axis of the rotation. You may support on a surface.
  • the sample adsorption region provided on the transfer film rotates in the reaction solution when the transfer film is supported by the transfer film support. Therefore, there is an effect that the sample and the reaction solution can be reacted uniformly.
  • the sample reaction method according to the present invention includes a step of rotating an adsorption structure including a sample adsorption region for adsorbing a sample in a reaction tank in which a reaction solution is stored, and the sample adsorption region includes: It is characterized by being provided on a surface covering the rotation axis of the rotation.
  • the sample storage chip according to the present invention includes a sample adsorption region for adsorbing a sample and a cover for covering the sample adsorption region, which are provided so as to cover one shaft.
  • a second sample reaction apparatus includes an adsorption structure having a sample adsorption region for adsorbing a sample, a reaction solution, and the adsorption structure so that the sample adsorption region is in contact with the reaction solution. It may be provided with a plurality of reaction tanks to be stored and a driving means for moving the adsorption structure to an arbitrary reaction tank.
  • a series of reaction (For example, blocking reaction in an antibody immune reaction, primary reaction) Antibody reaction, secondary antibody reaction, etc.) can be performed continuously.
  • a second sample reaction method includes a first reaction step of moving an adsorption structure having a sample adsorption region for adsorbing a sample into a first reaction tank in which a first reaction liquid is stored; It is characterized by including the 2nd reaction process of moving the said adsorption structure to the 2nd reaction tank in which the 2nd reaction liquid was stored after the 1st reaction process.
  • the reaction vessel may include a temperature adjusting means for adjusting the temperature of the reaction solution.
  • the temperature of the reaction solution can be adjusted to an appropriate temperature.
  • FIG. 3 shows an adsorption structure comprising a rod-shaped transfer film support according to an embodiment of the present invention
  • FIG. 3B shows an adsorption structure including a pipe-shaped transfer film support according to an embodiment of the present invention
  • FIG. 3C shows a screw according to an embodiment of the present invention
  • FIG. 3D shows an adsorption structure provided with a mesh-like transfer film support according to an embodiment of the present invention.
  • FIG. 4A is a cross-sectional view showing a variation of the coupling mode between the transfer film support and the transfer film according to the present invention
  • FIG. 4A is an adsorption structure in which the coupling portion is a slit according to an embodiment of the present invention.
  • FIG. 4 (b) shows an adsorbing structure in which the connecting portion is an adhesive according to an embodiment of the present invention
  • FIG. 4 (c) shows an embodiment of the present invention.
  • bond part is an insertion division part is shown. It is a schematic diagram explaining the surface in which the sample adsorption
  • FIG. 5 is a perspective view which shows the column-shaped adsorption
  • FIG. 5B shows a cross-sectional view for explaining the operation of the sample reaction apparatus provided with the adsorption structure. It is a schematic diagram explaining the surface in which the sample adsorption
  • FIG. 7 is a perspective view which shows the elliptical columnar adsorption structure which concerns on one Embodiment of this invention.
  • FIG. 7B shows a cross-sectional view for explaining the operation of the sample reaction apparatus provided with the adsorption structure.
  • FIG. 7B shows a cross-sectional view for explaining the operation of the sample reaction apparatus provided with the adsorption structure.
  • It is a perspective view showing a schematic structure of an adsorption structure provided with a cover concerning one embodiment of the present invention.
  • FIG. 10 shows schematic structure of the semi-columnar reaction tank which concerns on one Embodiment of this invention
  • FIG. 10 shows a schematic configuration of a cut-out polygonal wall-like reaction tank according to an embodiment of the present invention
  • FIG. 10C shows a reaction with oblique grooves according to an embodiment of the present invention.
  • the schematic structure of a tank is shown.
  • FIG. 12 is a top view illustrating a schematic configuration of a reaction layer provided with grooves or protrusions according to some embodiments of the present invention
  • FIG. 11A illustrates a groove obliquely on one side according to an embodiment of the present invention.
  • FIG. 12 is a top view illustrating a schematic configuration of a reaction layer provided with grooves or protrusions according to some embodiments of the present invention
  • FIG. 11A illustrates a groove obliquely on one side according to an embodiment of the present invention.
  • FIG. 11B shows a schematic configuration of a reaction tank in which grooves are obliquely provided on both sides according to an embodiment of the present invention. It is a schematic diagram which shows schematic structure of the sample reaction apparatus which concerns on one Embodiment of this invention, (a) of FIG. 12 is the outline of the sample reaction apparatus provided with the several reaction tank based on one Embodiment of this invention.
  • FIG. 12B is a top view showing a schematic configuration of the sample reaction apparatus. It is a photograph which shows the sample reaction apparatus which concerns on one Embodiment of this invention.
  • FIG. 15 is a photograph showing a result of contacting a sample with a reaction solution using a sample reaction apparatus according to the present invention or a conventional technique, and FIGS. 14 (a) to 14 (c) are drawings using the sample reaction apparatus according to the present invention. The result of contacting the sample with the reaction solution is shown.
  • FIG. 14D shows the result of reacting the sample with the reaction solution using the conventional technique.
  • the present invention provides a sample reaction apparatus.
  • the sample reaction apparatus is intended to be an apparatus for bringing a sample into contact with a reaction solution.
  • sample is used interchangeably with specimens, preparations in the art, and as used herein, “biological sample” or equivalent thereof is intended.
  • a “biological sample” is intended to be any preparation obtained from biological material as a source (eg, an individual, body fluid, cell line, tissue culture or tissue section).
  • tissue samples include body fluids (eg, blood, saliva, plaque, serum, plasma, urine, synovial fluid, and fluids) and tissue sources.
  • a preferred biological sample is a subject sample.
  • Preferred subject samples are skin lesions, sputum, pharyngeal mucus, nasal mucus, pus, or secretions obtained from the subject.
  • tissue sample intends a biological sample obtained from a tissue source. Methods for obtaining tissue biopsies and body fluids from mammals are well known in the art.
  • sample includes, in addition to the biological sample and the tissue sample, a protein sample, genomic DNA sample and / or extracted from the biological sample and the tissue sample.
  • a total RNA sample is also included.
  • reaction solution is intended to be a liquid reagent for reacting with a sample.
  • a reaction solution for use in the sample reaction apparatus according to the present invention for example, an antibody solution, an enzyme solution, a staining solution, a washing solution, a buffer solution, and the like can be used, but it is not particularly limited, depending on the sample to be used and the purpose. Conventional reagents well known in the art can be used.
  • the sample reaction apparatus when used for detecting a protein transferred to a transfer membrane by a Western blotting method using an antibody, the reaction solution is specifically a blocking buffer, and the above protein.
  • the reaction solution is specifically a blocking buffer, and the above protein.
  • Any of a primary antibody that binds, a secondary antibody that specifically binds to the primary antibody, a washing solution, and the like can be suitably used.
  • a solution containing the probe DNA can be used as a reaction solution.
  • the sample reaction apparatus can be used as a fully automatic protein staining apparatus.
  • a staining solution one that stains a protein itself, one that stains a sugar residue, a phosphate group, a nitro group, or the like of a protein can be used.
  • staining solutions include, but are not limited to, commercially available SYPRO Ruby (Invitrogen Corporation), MemCode Reversible Protein Stain Kit (TAKARA BIO INC.), GelCode Glycoprotein Staining Kit (Technochemical Co., Ltd.) )), Deep, Purple, Total, Protein, and Stain (GE Healthcare).
  • FIG. 1 is a cross-sectional view showing a schematic configuration of a sample reaction apparatus 100 according to an embodiment of the present invention.
  • FIG. 2 is a perspective view showing a schematic configuration of the sample reaction apparatus 100.
  • the sample reaction apparatus 100 according to this embodiment includes an adsorption structure 110, a reaction tank 120, and a rotation drive unit (drive means) 130.
  • the adsorption structure 110 includes a transfer film 115 and a transfer film support 114.
  • a sample adsorption region 112 that adsorbs the sample 111 is provided on the transfer film 115.
  • reaction liquid 121 is stored in the reaction tank 120.
  • the rotation drive unit 130 includes a motor 132 and a gear set 133 provided on the reaction tank 120 side, and a gear 134 coupled to the adsorption structure 110.
  • the sample reaction apparatus 100 includes the sample adsorption region 112 that adsorbs the sample 111, and the sample 111 adsorbed on the sample adsorption region 112 and the reaction solution 121 are brought into contact with each other.
  • the sample and the reaction liquid can be contacted uniformly over the entire area of the sample adsorption region. Therefore, according to the present invention, it is possible to uniformly react a sample with a reaction solution, which could not be achieved conventionally.
  • the sample adsorption region 112 may be any as long as it adsorbs the sample 111, and chemical, electrical, physical characteristics, or the like can be used for adsorption of the sample 111.
  • chemical, electrical, physical characteristics, or the like can be used for adsorption of the sample 111.
  • the transfer film 115 is determined depending on its polarity, hydrophobicity, or pore size.
  • a sample such as protein or DNA is adsorbed.
  • the transfer film 115 and the sample 111 may be firmly bonded by performing a process such as baking or UV cross-linking.
  • the adsorption structure 110 includes a transfer film 115 and a transfer film support 114 that supports the transfer film 115, and a sample adsorption region 112 is provided on the transfer film 115.
  • the adsorption structure 110 may be composed of a transfer film support 114 and a transfer film 115 as in this embodiment.
  • the adsorption structure 110 has a rod shape or a pipe shape.
  • a configuration in which the sample adsorption region 112 is provided on the surface may be used.
  • the method for adsorbing the sample onto the transfer film 115 is not particularly limited, and a well-known and commonly used method (for example, western blotting, dropping of cell lysate, etc.) can be used. Further, when the adsorption structure 110 has a rod shape or a pipe shape and the sample adsorption region 112 is provided on the surface, the adsorption structure 110 is made of a gel or the like on which the sample 111 is adsorbed. The sample 111 may be adsorbed on the adsorption structure 110 by rolling up.
  • the reaction tank 120 stores the reaction solution 121 and stores the adsorption structure 110 so that the sample adsorption region 112 is in contact with the reaction solution 121.
  • a temperature adjustment unit (temperature adjustment means) 123 such as a Peltier element is disposed on the bottom surface of the reaction tank 120, and the temperature adjustment unit 123 controls the temperature of the reaction liquid 121. You may do it.
  • the rotation drive unit (drive means) 130 may be anything that rotates the adsorption structure 110 around the rotation axis 131 in the reaction tank 120.
  • the rotation of the motor 132 is transmitted to the gear set 133, and the gear set 133 and the gear 134 are engaged with each other, whereby the adsorption structure 110 can be rotated.
  • the rotating shaft 131 is set in the reaction tank 120.
  • the gear set 133 is provided so that the gear set 133 and the gear 134 are engaged at a position where the center of the gear 134 coincides with the rotation shaft 131.
  • the motor 132 may be provided inside the adsorption structure 110.
  • the gear 134 is engaged with a gear set 133 fixed to the processing tank 120, thereby rotating the adsorption structure 110. .
  • the sample adsorption region 112 is provided on a surface covering the rotating shaft 131. Therefore, when the adsorption structure 110 rotates, the sample adsorption region 112 and the reaction liquid 121 can be brought into good contact with each other.
  • the surface covering the rotation shaft 131 is an outer surface of a surface provided so as to surround the rotation shaft 131.
  • FIG. 3 is a perspective view showing a variation of the structure of the transfer film support 114.
  • the transfer film support 114 may have a rod shape as shown in FIG. 3 (a) or a pipe-like structure as shown in FIG. 3 (b). A screw-like structure as shown in FIG. 3 or a mesh-like structure as shown in FIG.
  • the transfer film support 114 is formed into a screw-like structure as shown in FIG. 3C or a mesh-like structure as shown in FIG.
  • the transfer film 115 can be supported so as to have a gap between the two.
  • the reaction liquid 121 also flows on the back side of the transfer film 115 during the rotation. Therefore, the reaction liquid 121 does not stay, and the sample 111 and the reaction liquid 121 can be brought into contact more suitably.
  • FIG. 4 is a cross-sectional view showing variations in the coupling mode between the transfer film support 114 and the transfer film 115.
  • the transfer film 115 is coupled to a coupling portion 116 provided on the transfer film support 114.
  • the structure of the coupling portion 116 may be, for example, a slit that sandwiches the transfer film 115 as shown in FIG. 4A, or an adhesive that adheres the transfer film 115 as shown in FIG. 4B. (A magnet, a double-sided tape, or the like) or a divided portion that sandwiches the transfer film 115 with the entire transfer film support 114 as shown in FIG. 4C, and supports the transfer film 115.
  • the structure is not particularly limited as long as it can be structured.
  • FIG. 5 to 7 are schematic diagrams for explaining the surface 113 on which the sample adsorption region 112 is provided.
  • the surface 113 may be provided so as to cover the rotation shaft 131, but is preferably a surface parallel to the rotation shaft 131. If the surface 113 is a surface parallel to the rotation axis 131, the sample adsorption region 112 is provided on the surface 113 configured by a line orthogonal to the direction of movement by the rotation. Therefore, the moving speed accompanying the rotation of the sample 111 is equal for each line. Therefore, the sample 111 and the reaction liquid 121 can be contacted uniformly. Note that a plane parallel to a specific straight line refers to a plane that does not intersect the straight line.
  • the surface 113 is a cylindrical surface having a rotation axis 131 as a central axis 119, as shown in FIG.
  • each point on the surface 113 moves smoothly in the reaction solution 121 under the same conditions. Therefore, the sample 111 and the reaction liquid 121 can be contacted more uniformly.
  • the surface 113 is a cylindrical surface having a central axis 119 as an axis parallel to the rotation axis 131 as shown in FIG.
  • each point on the surface 113 moves smoothly in the reaction solution 121.
  • the reaction liquid 121 is agitated by the rotation of the surface 113, and the sample 111 and the reaction liquid 121 can be brought into contact more suitably.
  • the distance between the rotation shaft 131 and the central axis 119 of the surface 113 is preferably 50% or less of the radius of the surface 113.
  • the reaction liquid 121 can be stirred.
  • the distance is 50% or less of the radius, the reaction solution 121 and the sample 111 can be more reliably brought into contact with each other.
  • the surface 113 is an elliptical cylinder surface having a rotation axis 131 as a central axis 119, as shown in FIG. In this case, as shown in FIG. 7B, each point on the surface 113 moves smoothly in the reaction solution 121. Furthermore, the reaction liquid 121 is agitated by the rotation of the surface 113, and the sample 111 and the reaction liquid 121 can be brought into contact more suitably.
  • the flatness of the surface 113 is preferably 0.5 or less. When the flatness exceeds 0, the reaction liquid 121 can be stirred. When the flatness is 0.5 or less, the reaction solution 121 and the sample 111 can be contacted more uniformly.
  • the adsorption structure 110 may have a structure for protecting the sample adsorption region 112.
  • FIG. 8 is a perspective view illustrating a schematic configuration of the adsorption structure 110 including the cover 117.
  • FIG. 9 is a cross-sectional view illustrating a schematic configuration of the adsorption structure 110 including the cover 117.
  • the cover 117 is provided outside the transfer film 115 and covers the sample adsorption region 112. Therefore, the sample 111 adsorbed on the sample adsorption region 112 can be prevented from coming into contact with other articles. Therefore, the adsorption structure 110 can be stored while the sample 111 is adsorbed.
  • Such an adsorption structure 110 provided with the cover 117 can be reused for diagnosing a disease by storing it in a refrigerated state as a 2DE chip in advance and setting it in a device when it becomes ill. . That is, the present invention also provides a sample storage chip having the above-described configuration.
  • the cover 117 also allows the reaction solution 121 to pass therethrough.
  • a cover 117 is not limited to this, but may have an opening 118 so that the reaction liquid 121 can enter and exit from the opening 118.
  • the opening 118 may be cut as shown in FIG. 8 (that is, a part of the cover 117 is cut out), and is mesh-shaped (that is, a part of the cover 117 is mesh-shaped). May be.
  • the reaction solution 121 is passed through the opening 118. Reaches the sample adsorption region 112, so that the sample 111 and the reaction liquid 121 can be reacted. If the cover 117 does not pass through the reaction solution 121, the cover 117 may be removed before the adsorption structure 110 is stored in the reaction tank 120.
  • FIG. 10 is a perspective view showing a variation of the reaction tank 120.
  • the inner surface of the reaction tank 120 is a semi-cylindrical surface, a semi-cylindrical surface shape, or a central axis parallel to the length direction of a hexagonal column or more polygonal column. It is preferable that the shape is a wall surface when cut by a plane passing through (excluding the cut surface, ie, a shape approximating a semi-cylindrical surface).
  • the adsorption structure 110 can be suitably accommodated by setting the central axis of the semi-cylindrical surface and the rotation axis of the adsorption structure 110 to the same position.
  • the adsorption structure 110 can be accommodated in the reaction tank 120 with a slight gap.
  • the reaction liquid 121 can be brought into contact with a wide range of the adsorption structure 110 with a small liquid volume, and the required amount of the reaction liquid 121 can be reduced. it can.
  • the inner surface of the reaction tank 120 is not a semi-cylindrical surface but the above wall surface shape, an effect close to the semi-cylindrical surface can be obtained.
  • FIG. 11 is a top view of the reaction vessel 120 provided with such grooves or protrusions.
  • FIG. 11A shows a configuration in which grooves or protrusions are provided on one side of the reaction tank 120
  • FIG. 11B shows a configuration in which grooves or protrusions are provided on both sides of the reaction tank 120.
  • the reaction liquid 121 flows along the direction of the rotation due to the rotation of the adsorption structure 110, but the direction of the flow can be changed obliquely along the grooves or protrusions (in FIG. 11). Arrow direction). Therefore, since the reaction liquid 121 is stirred without the flow direction of the reaction liquid 121 being unidirectional, the sample 111 and the reaction liquid 121 can be more uniformly brought into contact with each other.
  • the sample reaction apparatus 100 according to the present embodiment can be used for the transfer film 115 to which the protein separated by two-dimensional electrophoresis is transferred, for example. Since such a transfer film has a large area, it has been difficult for the transfer film to uniformly react with the reaction liquid 121 by the conventional technique. For the analysis of two-dimensional electrophoresis, it is particularly important that the concentration of each part is uniform, so that the sample reaction apparatus 100 according to the present embodiment can be used particularly suitably.
  • FIG. 12 is a schematic diagram showing a schematic configuration of a sample reaction apparatus 200 according to another embodiment of the present invention, in which (a) of FIG. 12 shows a sectional view and (b) of FIG. 12 shows a top view. .
  • the sample reaction apparatus 200 includes an adsorption structure 210, reaction tanks 220, 222, 224, and 226, a waste liquid tank 228, and a drive unit (drive means) 230.
  • a drive unit drive means
  • Reaction liquids 221, 223, 225, and 227 are stored in the reaction tanks 220, 222, 224, and 226, respectively.
  • the drive unit 230 includes a motor 232, a gear set 233, and an arm 234.
  • the arm 234 moves the adsorption structure 210 to an arbitrary reaction tank.
  • the arm 234 may be configured using, for example, a well-known and common robot arm technique.
  • the sample reaction apparatus 200 can be used for a series of reactions of (i) blocking treatment, (ii) reaction with a primary antibody, (iii) washing, and (iv) reaction with a secondary antibody.
  • the reaction solution 221 is a blocking buffer solution
  • the reaction solution 223 is a blocking buffer solution containing a primary antibody
  • the reaction solution 225 is a blocking reaction solution containing a secondary antibody
  • the reaction solution 227 is a washing solution. It can be.
  • a cleaning liquid as the reaction liquid 227 may be sequentially added to the reaction tank 226, for example, to the part to be added 229.
  • a waste liquid tank 228 for collecting the overflowing reaction liquid 227 may be provided adjacent to the reaction tank 226 as shown in FIG.
  • the addition means 230 of the reaction liquid 227 for example, a tube with a valve well known and commonly used by those skilled in the art can be used.
  • sample reaction apparatus 200 As described above, if the sample reaction apparatus 200 according to this embodiment is used, a series of reactions can be performed sequentially, so that complicated processes such as detection of Western blotting results can be automated.
  • a photograph of an example of the sample reactor 200 is shown in FIG.
  • the sample reaction apparatus 200 includes a plurality of reaction vessels, and an object of the present invention is to automate a series of reactions by sequentially moving an adsorption structure that adsorbs a sample in each reaction vessel.
  • an object of the present invention is to automate a series of reactions by sequentially moving an adsorption structure that adsorbs a sample in each reaction vessel.
  • the transfer film was placed on a screener blotter (Samplatech), 85 ⁇ l of each sample was applied, and allowed to stand for 40 minutes to adsorb the sample onto the transfer film.
  • the transfer membrane is treated with a blocking buffer (TBS-T containing 1.5% skim milk (0.05% Tween 20, 50 mM Tris (24.2 g / 4 L), 150 mM NaCl (35 g / 4 L))) for 30 minutes. Reacted.
  • TBS-T a blocking buffer
  • skim milk 0.05% Tween 20, 50 mM Tris (24.2 g / 4 L), 150 mM NaCl (35 g / 4 L)
  • reaction process with primary antibody After the blocking treatment, the transfer membrane was reacted with a primary antibody diluted with a blocking buffer (anti-carbonic anhydrobrom from Bovine Erythrocytes rabbit antibody (NE023 / 7S, Nordic Immunological Lab.)).
  • a blocking buffer anti-carbonic anhydrobrom from Bovine Erythrocytes rabbit antibody (NE023 / 7S, Nordic Immunological Lab.)
  • reaction process with secondary antibody After washing, the transfer membrane was reacted with a secondary antibody (Alexa Fluor 647 goat anti-rabbit IgG (H + L) (A21244, Invitrogen Co.)) diluted with a blocking buffer.
  • a secondary antibody Alexa Fluor 647 goat anti-rabbit IgG (H + L) (A21244, Invitrogen Co.)
  • the band on the obtained transfer film was detected by a fluorescence scanner (Typhoon TRIO +, GE Healthcare).
  • Example 1 The blocking treatment, the reaction with the primary antibody, the washing, and the reaction with the secondary antibody described above are shown in FIG. 12 in the blocking treatment, the reaction with the primary antibody, the washing, and the reaction with the secondary antibody.
  • the reaction was carried out using a sample reaction apparatus according to the present invention equipped with a reaction tank for each reaction. A transfer film support having a diameter of 19 mm and having a screw shape as shown in FIG. 3C was used.
  • a reaction vessel for blocking treatment reaction with the primary antibody, and reaction with the secondary antibody, a semi-cylindrical shape having a diameter of 20 mm is used, and as a reaction vessel for washing, a semi-cylindrical shape having a diameter of 22 mm is used. The thing of was used. All reactions were performed under conditions of 20 ° C. and 70% humidity.
  • the support for supporting the transfer film is rotated at a speed of 18 revolutions / minute in the reaction tank for each. It went by.
  • the amount of blocking buffer in the blocking step was 5 ml.
  • the reaction solution in the reaction step with the primary antibody or the secondary antibody was prepared by diluting 1 ⁇ l of the above antibody so that the total amount became 5 ml, and the reaction time was 60 minutes.
  • Example 2 The reaction was carried out in the same manner as in Example 1 except that the reaction time in the reaction step with the primary antibody or the secondary antibody was 15 minutes. The result is shown in FIG.
  • Example 3 As in Example 1, except that the reaction solution in the reaction step with the primary antibody or the secondary antibody was prepared by diluting 1 ⁇ l of the above antibody to a total volume of 1.23 ml, and setting the reaction time to 15 minutes. Reaction was performed. The results are shown in FIG.
  • the amount of blocking buffer in the blocking step was 25 ml.
  • the reaction solution in the reaction step with the primary antibody or the secondary antibody was prepared by diluting 5 ⁇ l of the above antibody so that the total amount became 5 ml, and the reaction time was 60 minutes.
  • the amount of the cleaning liquid in the cleaning process was 25 ml. The result is shown in FIG.
  • the band obtained by the conventional method has a thin central portion (see (d) of FIG. 14).
  • the sample reaction apparatus according to the present invention is used, the whole color is uniformly developed (see FIGS. 14A to 14C).
  • the adsorption structure having the sample adsorption region for adsorbing the sample is rotated in the reaction tank for storing the reaction solution, so that the sample can be reacted uniformly with the reaction solution. it can.
  • the present invention can be used in the fields of various experimental devices, inspection devices, measuring devices, and the like.

Abstract

Disclosed is a sample reactor (100) comprising an adsorption structure (110), which is equipped with a sample adsorption zone (112) that adsorbs a sample (111), a reaction tank (120) that holds a reaction solution (121) and accommodates the adsorption structure (110) so that the sample adsorption zone (112) is in contact with the reaction solution (121), and a rotary drive unit (130) that rotates the adsorption structure (110) inside the reaction tank (120), wherein the sample adsorption zone (112) is arranged on a surface that covers the rotational axis of said rotation. In this way, the sample reactor which is provided causes a sample that has been transferred to a transfer film to uniformly react with the reaction solution.

Description

サンプル反応装置およびサンプル反応方法Sample reaction apparatus and sample reaction method
　本発明は、サンプルを反応液と反応させるために用いる装置および方法に関するものである。 The present invention relates to an apparatus and method used for reacting a sample with a reaction solution.
　ヒトゲノムプロジェクトが終了した後、プロテオーム研究が盛んに行われている。「プロテオーム」とは、特定の細胞、器官または臓器の中で翻訳生産されているタンパク質全体が意図され、その研究としてはタンパク質のプロファイリングなどが挙げられる。 After the Human Genome Project has been completed, proteome research has been actively conducted. The term “proteome” is intended to mean a whole protein produced by translation in a specific cell, organ or organ, and its research includes protein profiling.
　タンパク質をプロファイリングする手法の１つとして最も用いられているものが、タンパク質の電気泳動、特に２次元電気泳動である。タンパク質は、電荷および分子量の独特の性質を有しているので、多数のタンパク質の混合物であるプロテオームから電荷のみまたは分子量のみに依存して個々のタンパク質を各成分に分離するよりも、両者を組み合わせることにより、より多くのタンパク質を高分解能にて分離することができる。 One of the most widely used methods for profiling proteins is protein electrophoresis, particularly two-dimensional electrophoresis. Because proteins have unique properties of charge and molecular weight, they combine a proteome, which is a mixture of many proteins, rather than separating individual proteins into their components based on charge alone or molecular weight alone. As a result, more proteins can be separated with high resolution.
　電気泳動によってタンパク質は電荷および／または分子量によって分離されるが、このような物理的性質のみで分離したタンパク質についてその分離位置から生物学的性質を特定することは困難である。また、タンパク質は合成された後にリン酸化などの化学的修飾（翻訳後修飾）を受けることによりその機能が制御されることが知られている。電気泳動のみではこのような翻訳後修飾に関する情報を得ることは困難である。 Proteins are separated by charge and / or molecular weight by electrophoresis, but it is difficult to specify biological properties from the separation position of proteins separated only by such physical properties. In addition, it is known that the function of a protein is controlled by being subjected to chemical modification (post-translational modification) such as phosphorylation after being synthesized. It is difficult to obtain information on such post-translational modifications by electrophoresis alone.
　ウェスタンブロッティング法は電気泳動によって分離されたスラブゲル中のタンパク質を膜に転写する方法である。ウェスタンブロッティング法によりタンパク質が転写された膜上に特定の抗体をオーバーレイすれば、抗原抗体反応に基づいてタンパク質をある程度特定することが可能となる。また、翻訳後修飾の１つであるリン酸化について、タンパク質が転写された膜に抗リン酸化タンパク質抗体をオーバーレイすることにより、リン酸化の有無、リン酸化部位の相違を検出することが可能となる。 Western blotting is a method of transferring proteins in a slab gel separated by electrophoresis to a membrane. If a specific antibody is overlaid on the membrane onto which the protein has been transferred by Western blotting, the protein can be specified to some extent based on the antigen-antibody reaction. In addition, regarding phosphorylation, which is one of post-translational modifications, it is possible to detect the presence or absence of phosphorylation and the difference in phosphorylation sites by overlaying an anti-phosphorylated protein antibody on the membrane onto which the protein has been transferred. .
　タンパク質が転写された膜上に特定の抗体をオーバーレイすることは、通常、転写膜を、プラスチック容器やビニールバックなどの中で、ブロッキング緩衝液、一次抗体、洗浄液、および二次抗体のそれぞれと反応させることによって行われる。これらの操作は、通常、手作業で行われる。このとき、上記反応を促進させるために、上記プラスチック容器またはビニールバッグを、往復運動または回転運動させることも行われる。また、特許文献１に記載のような検出法も開発されている。 Overlaying a specific antibody on the membrane onto which the protein has been transferred usually involves reacting the transfer membrane with each of blocking buffer, primary antibody, washing solution, and secondary antibody in a plastic container or plastic bag, etc. Is done by letting These operations are usually performed manually. At this time, in order to promote the reaction, the plastic container or the plastic bag is reciprocated or rotated. A detection method as described in Patent Document 1 has also been developed.
日本国公開特許公報「特開２００４－１６３１４６号公報（公開日：平成１６年６月１０日）」Japanese Patent Publication “JP 2004-163146 A (publication date: June 10, 2004)”
　しかしながら、従来技術では、転写膜に転写されたサンプルと、反応液とを均一に反応させることは困難である。 However, with the conventional technique, it is difficult to uniformly react the sample transferred to the transfer film and the reaction solution.
　本発明は、上記課題に鑑みてなされたものであり、転写膜に転写されたサンプルと、反応液とを均一に反応させるサンプル反応装置を提供することを主たる目的とする。 The present invention has been made in view of the above problems, and a main object of the present invention is to provide a sample reaction apparatus that uniformly reacts a sample transferred to a transfer film and a reaction solution.
　本発明に係るサンプル反応装置は、上記課題を解決するために、サンプルを吸着するサンプル吸着領域を備えている吸着構造体と、反応液を貯め、該サンプル吸着領域が該反応液と接するように該吸着構造体を収納する反応槽と、該吸着構造体を該反応槽内で回転させる駆動手段とを備えており、該サンプル吸着領域は、該回転の回転軸を覆う面上に設けられていることを特徴としている。 In order to solve the above problems, a sample reaction apparatus according to the present invention stores an adsorption structure having a sample adsorption region for adsorbing a sample, a reaction solution, and the sample adsorption region is in contact with the reaction solution. A reaction tank that houses the adsorption structure; and a drive unit that rotates the adsorption structure in the reaction tank. The sample adsorption region is provided on a surface that covers the rotation axis of the rotation. It is characterized by being.
　上記の構成によれば、上記サンプル吸着領域は、上記反応液内で、水車のように途切れなく回転運動する。それゆえ、従来技術のように往復運動させた場合に比べ、上記サンプルと上記反応液とを均一に反応させることができ、従来技術のように、上記反応槽ごと回転させた場合と比べ、該回転槽の運動に伴う該反応液の偏りを避けることができるので、上記サンプルと上記反応液とを均一に反応させることができるという効果を奏する。 According to the above configuration, the sample adsorption region rotates in a continuous manner like a water wheel in the reaction solution. Therefore, compared to the case where the sample is reciprocated as in the prior art, the sample and the reaction solution can be reacted uniformly, and compared with the case where the entire reaction vessel is rotated as in the conventional technique, Since unevenness of the reaction solution accompanying the movement of the rotating tank can be avoided, there is an effect that the sample and the reaction solution can be reacted uniformly.
　上記サンプル反応装置では、上記回転軸を覆う面が、上記回転軸と平行な面であることが好ましい。 In the sample reaction apparatus, the surface covering the rotation axis is preferably a surface parallel to the rotation axis.
　上記の構成によれば、上記サンプル吸着領域が、回転軸と平行な面上に設けられている。そのため、上記サンプル吸着領域は、上記回転軸を中心とする回転に対して、直交する線から構成される。そのため、上記各線上のサンプルは、上記回転に伴う移動速度が等しく、均一にサンプルと反応液とを接触させることができる。 According to the above configuration, the sample adsorption region is provided on a plane parallel to the rotation axis. Therefore, the sample adsorption region is composed of a line orthogonal to the rotation about the rotation axis. Therefore, the samples on each line have the same moving speed with the rotation, and the sample and the reaction liquid can be uniformly contacted.
　上記サンプル反応装置では、上記回転軸を覆う面が、円柱面、または楕円柱面であることが好ましい。 In the sample reaction apparatus, the surface covering the rotation axis is preferably a cylindrical surface or an elliptical cylindrical surface.
　上記の構成によれば、上記反応液内における上記サンプル吸着領域の運動をより滑らかにすることができるため、上記サンプルと、該反応液との反応のむらを抑制することができる。 According to the above configuration, since the movement of the sample adsorption region in the reaction solution can be made smoother, uneven reaction between the sample and the reaction solution can be suppressed.
　上記サンプル反応装置では、上記吸着構造体が、上記サンプル吸着領域が設けられている転写膜と、該転写膜を支持する転写膜支持体とをさらに備えていてもよい。 In the sample reaction apparatus, the adsorption structure may further include a transfer film provided with the sample adsorption region and a transfer film support that supports the transfer film.
　上記の構成によれば、上記転写膜上に上記サンプル吸着領域が設けられているため、上記サンプルを容易に上記吸着構造体に吸着させることができるという効果を奏する。 According to the above configuration, since the sample adsorption region is provided on the transfer film, there is an effect that the sample can be easily adsorbed to the adsorption structure.
　上記サンプル反応装置では、上記転写膜支持体と上記転写膜との間に空隙が設けられていることが好ましい。また、上記転写膜支持体が、上記駆動手段による回転の回転軸に平行な直線に斜めに交わる方向に沿った溝が設けられていることが好ましい。 In the sample reaction apparatus, it is preferable that a gap is provided between the transfer film support and the transfer film. Further, it is preferable that the transfer film support is provided with a groove along a direction obliquely intersecting with a straight line parallel to a rotation axis of rotation by the driving unit.
　上記の構成によれば、上記反応液が、上記転写膜の裏側を通過することが可能となる。そのため、上記反応液の流動が容易となり、上記サンプルと該反応液とをより均一に反応させることができるという効果を奏する。 According to the above configuration, the reaction solution can pass through the back side of the transfer film. Therefore, the flow of the reaction liquid becomes easy, and there is an effect that the sample and the reaction liquid can be reacted more uniformly.
　上記サンプル反応装置では、上記吸着構造体が、上記サンプル吸着領域を覆うカバーをさらに備えていてもよい。 In the sample reaction device, the adsorption structure may further include a cover that covers the sample adsorption region.
　上記の構成によれば、上記サンプル吸着領域の外側にカバーが備えられているので、上記吸着構造体に上記サンプルを吸着させたまま保存することができるという効果を奏する。 According to the above configuration, since the cover is provided outside the sample adsorption region, there is an effect that the sample can be stored while being adsorbed on the adsorption structure.
　上記サンプル反応装置では、上記カバーが、上記反応液が透過させることが好ましい。 In the sample reaction apparatus, it is preferable that the cover allows the reaction solution to pass through.
　上記の構成によれば、上記カバーが、上記反応液を透過するため、該カバーを付けたまま、上記吸着構造体を上記反応槽に収納させ、回転させることにより、上記サンプルと上記反応液とを反応させることができるという効果を奏する。 According to said structure, since the said cover permeate | transmits the said reaction liquid, the said adsorption | suction structure is accommodated in the said reaction tank with this cover attached, and by rotating, the said sample, the said reaction liquid, There is an effect that can be reacted.
　上記サンプル反応装置では、上記反応槽の内面が、半円柱面状、または六角柱以上の多角柱を長さ方向に対して平行な中心軸を通る面で切断したときの壁面状であることが好ましい。 In the sample reaction apparatus, the inner surface of the reaction vessel may be a semi-cylindrical surface shape or a wall surface shape when a polygonal column equal to or higher than a hexagonal column is cut by a plane passing through a central axis parallel to the length direction. preferable.
　上記の構成によれば、上記反応槽が、半円柱面か、または半円柱面に近い形状を有しているため、回転する上記吸着構造体をスペースの無駄なく収納することができ、これにより、必要となる上記反応液の液量を抑えられるという効果を奏する。 According to said structure, since the said reaction tank has a shape close | similar to a semi-cylindrical surface or a semi-cylindrical surface, the said adsorption | suction structure which rotates can be accommodated without waste of space, thereby There is an effect that the required amount of the reaction solution can be suppressed.
　上記サンプル反応装置では、上記反応槽の内面に、上記回転軸に平行な直線に斜めに交わる方向に沿った溝または突起が設けられていてもよい。 In the sample reaction apparatus, a groove or a protrusion along a direction obliquely intersecting with a straight line parallel to the rotation axis may be provided on the inner surface of the reaction vessel.
　上記の構成によれば、上記吸着構造体の回転により生じる上記反応液の該回転の方向に沿った流動が、該回転軸に平行な直線に斜めに交わる方向に沿った溝または突起によって、斜めに方向転換され、該反応液が、該回転の方向だけでなく、斜めの方向にも流動され、該反応液がより攪拌されるため、上記サンプルと該反応液とをより均一に反応させることができるという効果を奏する。 According to the above configuration, the flow of the reaction liquid generated by the rotation of the adsorption structure along the direction of the rotation is slanted by the grooves or protrusions along the direction that obliquely intersects the straight line parallel to the rotation axis. Since the reaction solution is flowed not only in the direction of rotation but also in an oblique direction and the reaction solution is further stirred, the sample and the reaction solution are reacted more uniformly. There is an effect that can be.
　上記サンプル反応装置では、上記反応槽に、上記反応液を添加する添加手段と、該反応槽から流出する該反応液を受ける廃液槽とをさらに備えていてもよい。 In the sample reaction apparatus, the reaction tank may further include an adding means for adding the reaction liquid and a waste liquid tank for receiving the reaction liquid flowing out from the reaction tank.
　上記の構成によれば、未反応の上記反応液を新たに添加し、反応済みの該反応液を回収することができるので、上記サンプルと未反応の該反応液とを反応させることができるという効果を奏する。 According to the above configuration, since the unreacted reaction solution can be newly added and the reacted reaction solution can be recovered, the sample and the unreacted reaction solution can be reacted. There is an effect.
　上記サンプル反応装置では、上記サンプルがタンパク質であり、上記反応液が、該タンパク質の抗体を含んでいるものであってもよい。 In the sample reaction apparatus, the sample may be a protein, and the reaction solution may contain an antibody of the protein.
　上記の構成によれば、二次元電気泳動によって分離されたタンパク質のように、均一に抗体と反応させることが重要なサンプルを均一に該抗体と反応させることができるという効果を奏する。 According to the above configuration, it is possible to uniformly react a sample that is important to be reacted with an antibody uniformly, such as a protein separated by two-dimensional electrophoresis.
　上記サンプル反応装置では、複数の上記反応槽を備えており、上記駆動手段が、上記吸着構造体を任意の該反応槽に移動させるものであってもよい。 The sample reaction apparatus may include a plurality of the reaction vessels, and the driving unit may move the adsorption structure to any reaction vessel.
　上記の構成によれば、上記反応槽が複数であるので、一連の反応（例えば、抗体免疫反応における、ブロッキング反応、一次抗体反応、および二次抗体反応等）を連続して行うことができるという効果を奏する。 According to the above configuration, since there are a plurality of reaction vessels, a series of reactions (for example, a blocking reaction, a primary antibody reaction, a secondary antibody reaction, etc. in an antibody immune reaction) can be performed continuously. There is an effect.
　上記サンプル反応装置では、上記サンプルがタンパク質であり、上記複数の反応槽が、ブロッキング緩衝液を貯める第１反応槽、該タンパク質に結合する一次抗体を含む反応液を貯める第２反応槽、該一次抗体に結合する二次抗体を含む反応液を貯める第３反応槽、洗浄液を貯める第４反応槽を含んでいてもよい。 In the sample reaction apparatus, the sample is protein, and the plurality of reaction tanks are a first reaction tank that stores a blocking buffer, a second reaction tank that stores a reaction liquid containing a primary antibody that binds to the protein, and the primary reaction tank. You may include the 3rd reaction tank which stores the reaction liquid containing the secondary antibody couple | bonded with an antibody, and the 4th reaction tank which stores a washing | cleaning liquid.
　上記の構成によれば、二次元電気泳動によって分離されたタンパク質のウェスタンブロッティング後の上記サンプルの検出を好適に行うことができる。 According to the above configuration, it is possible to suitably detect the sample after Western blotting of proteins separated by two-dimensional electrophoresis.
　本発明に係るサンプル反応装置はまた、サンプルを吸着する転写膜を支持する転写膜支持体と、反応液を貯め、該転写膜支持体が保持する転写膜が該反応液と接するように該転写膜支持体を収納する反応槽と、該転写膜支持体を該反応槽内で回転させる回転手段とを備えており、該転写膜支持体は、該転写膜を、該回転の回転軸を覆う面上に支持するものであってもよい。 The sample reaction apparatus according to the present invention also includes a transfer film support that supports a transfer film that adsorbs a sample and a reaction liquid, and the transfer film held by the transfer film support is in contact with the reaction liquid. A reaction tank containing the film support; and a rotating means for rotating the transfer film support in the reaction tank. The transfer film support covers the rotation axis of the rotation. You may support on a surface.
　上記の構成によれば、上記転写膜上に設けられたサンプル吸着領域は、該転写膜が上記転写膜支持体に支持されることにより、上記反応液内で回転運動する。それゆえ、上記サンプルと上記反応液とを均一に反応させることができるという効果を奏する。 According to the above configuration, the sample adsorption region provided on the transfer film rotates in the reaction solution when the transfer film is supported by the transfer film support. Therefore, there is an effect that the sample and the reaction solution can be reacted uniformly.
　本発明に係るサンプル反応方法は、サンプルを吸着するサンプル吸着領域を備えている吸着構造体を、反応液が貯められた反応槽内で回転させる工程を包含しており、該サンプル吸着領域は、該回転の回転軸を覆う面上に設けられていることを特徴としている。 The sample reaction method according to the present invention includes a step of rotating an adsorption structure including a sample adsorption region for adsorbing a sample in a reaction tank in which a reaction solution is stored, and the sample adsorption region includes: It is characterized by being provided on a surface covering the rotation axis of the rotation.
　上記の構成によれば、本発明に係るサンプル反応装置と同等の効果を奏することができる。 According to the above configuration, it is possible to achieve the same effect as the sample reaction apparatus according to the present invention.
　本発明に係るサンプル保存用チップは、一つの軸を覆うように設けられている、サンプルを吸着するサンプル吸着領域と、該サンプル吸着領域を覆うカバーとを備えていることを特徴としている。 The sample storage chip according to the present invention includes a sample adsorption region for adsorbing a sample and a cover for covering the sample adsorption region, which are provided so as to cover one shaft.
　上記の構成によれば、本発明に係るサンプル反応装置に組み込むための吸着構造体であって、サンプルが吸着されたまま保存可能なサンプル保存用チップを提供することができる。 According to the above configuration, it is possible to provide a sample storage chip that is an adsorption structure to be incorporated into the sample reaction apparatus according to the present invention and can be stored while the sample is adsorbed.
　本発明に係る第２のサンプル反応装置は、サンプルを吸着するサンプル吸着領域を備えている吸着構造体と、反応液を貯め、該サンプル吸着領域が該反応液と接するように該吸着構造体を収納する複数の反応槽と、該吸着構造体を任意の該反応槽に移動させる駆動手段とを備えているものであってもよい。 A second sample reaction apparatus according to the present invention includes an adsorption structure having a sample adsorption region for adsorbing a sample, a reaction solution, and the adsorption structure so that the sample adsorption region is in contact with the reaction solution. It may be provided with a plurality of reaction tanks to be stored and a driving means for moving the adsorption structure to an arbitrary reaction tank.
　上記の構成によれば、複数の上記反応槽を備え、上記吸着構造体を該反応槽間で移動させる駆動手段を備えているので、一連の反応（例えば、抗体免疫反応における、ブロッキング反応、一次抗体反応、および二次抗体反応等）を連続して行うことができるという効果を奏する。 According to said structure, since it is equipped with the said several reaction tank and the drive means which moves the said adsorption | suction structure between this reaction tank, a series of reaction (For example, blocking reaction in an antibody immune reaction, primary reaction) Antibody reaction, secondary antibody reaction, etc.) can be performed continuously.
　本発明に係る第２のサンプル反応方法は、サンプルを吸着するサンプル吸着領域を備えている吸着構造体を、第１反応液が貯められた第１反応槽内に移動させる第１反応工程と、第１反応工程の後に、上記吸着構造体を、第２反応液が貯められた第２反応槽に移動させる第２反応工程とを包含することを特徴としている。 A second sample reaction method according to the present invention includes a first reaction step of moving an adsorption structure having a sample adsorption region for adsorbing a sample into a first reaction tank in which a first reaction liquid is stored; It is characterized by including the 2nd reaction process of moving the said adsorption structure to the 2nd reaction tank in which the 2nd reaction liquid was stored after the 1st reaction process.
　上記の構成によれば、本発明に係る第２のサンプル反応装置と同等の効果を奏することができる。 According to the above configuration, the same effects as those of the second sample reaction apparatus according to the present invention can be achieved.
　本発明に係るサンプル反応装置では、上記反応槽が、上記反応液の温度を調節する温度調節手段を備えていてもよい。 In the sample reaction apparatus according to the present invention, the reaction vessel may include a temperature adjusting means for adjusting the temperature of the reaction solution.
　上記の構成によれば、反応液の温度を適切な温度に調節することができる。 According to the above configuration, the temperature of the reaction solution can be adjusted to an appropriate temperature.
　本発明の他の目的、特徴、および優れた点は、以下に示す記載によって十分分かるであろう。また、本発明の利点は、添付図面を参照した次の説明で明白になるであろう。 Other objects, features, and superior points of the present invention will be fully understood from the following description. The advantages of the present invention will become apparent from the following description with reference to the accompanying drawings.
本発明の一実施形態に係るサンプル反応装置の概略構成を示す断面図である。It is sectional drawing which shows schematic structure of the sample reaction apparatus which concerns on one Embodiment of this invention. 本発明の一実施形態に係るサンプル反応装置の概略構成を示す斜視図である。It is a perspective view which shows schematic structure of the sample reaction apparatus which concerns on one Embodiment of this invention. 本発明に係る転写膜支持体の構造のバリエーションを示す斜視図であり、図３の（ａ）は、本発明の一実施形態に係る、棒状の転写膜支持体を備える吸着構造体を示し、図３の（ｂ）は、本発明の一実施形態に係る、パイプ状の転写膜支持体を備える吸着構造体を示し、図３の（ｃ）は、本発明の一実施形態に係る、スクリュー状の転写膜支持体を備える吸着構造体を示し、図３の（ｄ）は、本発明の一実施形態に係る、メッシュ状の転写膜支持体を備える吸着構造体を示す。It is a perspective view showing a variation of the structure of the transfer film support according to the present invention, FIG. 3 (a) shows an adsorption structure comprising a rod-shaped transfer film support according to an embodiment of the present invention, FIG. 3B shows an adsorption structure including a pipe-shaped transfer film support according to an embodiment of the present invention, and FIG. 3C shows a screw according to an embodiment of the present invention. FIG. 3D shows an adsorption structure provided with a mesh-like transfer film support according to an embodiment of the present invention. 本発明に係る転写膜支持体と転写膜との結合様式のバリエーションを示す断面図であり、図４の（ａ）は、本発明の一実施形態に係る、結合部がスリットである吸着構造体を示し、図４の（ｂ）は、本発明の一実施形態に係る、結合部が接着物である吸着構造体を示し、図４の（ｃ）は、本発明の一実施形態に係る、結合部が挟み込み分割部である吸着構造体を示す。FIG. 4A is a cross-sectional view showing a variation of the coupling mode between the transfer film support and the transfer film according to the present invention, and FIG. 4A is an adsorption structure in which the coupling portion is a slit according to an embodiment of the present invention. FIG. 4 (b) shows an adsorbing structure in which the connecting portion is an adhesive according to an embodiment of the present invention, and FIG. 4 (c) shows an embodiment of the present invention. The adsorption | suction structure whose coupling | bond part is an insertion division part is shown. 本発明の一実施形態に係るサンプル吸着領域が設けられる面を説明する模式図であり、図５の（ａ）は、本発明の一実施形態に係る円柱状の吸着構造体を示す斜視図を示し、図５の（ｂ）は、上記吸着構造体を備えたサンプル反応装置の動作を説明する断面図を示す。It is a schematic diagram explaining the surface in which the sample adsorption | suction area | region which concerns on one Embodiment of this invention is provided, (a) of FIG. 5 is a perspective view which shows the column-shaped adsorption | suction structure concerning one Embodiment of this invention. FIG. 5B shows a cross-sectional view for explaining the operation of the sample reaction apparatus provided with the adsorption structure. 本発明の一実施形態に係るサンプル吸着領域が設けられる面を説明する模式図であり、図６の（ａ）は、本発明の一実施形態に係る中心軸と回転軸とがずれている吸着構造体を示す斜視図を示し、図６の（ｂ）は、上記吸着構造体を備えたサンプル反応装置の動作を説明する断面図を示す。It is a schematic diagram explaining the surface in which the sample adsorption | suction area | region which concerns on one Embodiment of this invention is provided, (a) of FIG. 6 is the adsorption | suction with which the center axis | shaft and rotational axis which concern on one Embodiment of this invention have shifted | deviated. The perspective view which shows a structure is shown, (b) of FIG. 6 shows sectional drawing explaining operation | movement of the sample reaction apparatus provided with the said adsorption structure. 本発明の一実施形態に係るサンプル吸着領域が設けられる面を説明する模式図であり、図７の（ａ）は、本発明の一実施形態に係る楕円柱状の吸着構造体を示す斜視図を示し、図７の（ｂ）は、上記吸着構造体を備えたサンプル反応装置の動作を説明する断面図を示す。It is a schematic diagram explaining the surface in which the sample adsorption | suction area | region which concerns on one Embodiment of this invention is provided, (a) of FIG. 7 is a perspective view which shows the elliptical columnar adsorption structure which concerns on one Embodiment of this invention. FIG. 7B shows a cross-sectional view for explaining the operation of the sample reaction apparatus provided with the adsorption structure. 本発明の一実施形態に係る、カバーを備えた吸着構造体の概略構成を示す斜視図である。It is a perspective view showing a schematic structure of an adsorption structure provided with a cover concerning one embodiment of the present invention. 本発明の一実施形態に係る、カバーを備えた吸着構造体を備えたサンプル反応装置の概略構成を示す断面図である。It is sectional drawing which shows schematic structure of the sample reaction apparatus provided with the adsorption | suction structure provided with the cover based on one Embodiment of this invention. 本発明に係る反応槽のバリエーションを示す斜視図であり、図１０の（ａ）は、本発明の一実施形態に係る半円柱状の反応槽の概略構成を示し、図１０の（ｂ）は、本発明の一実施形態に係る切断された多角柱の壁面状の反応槽の概略構成を示し、図１０の（ｃ）は、本発明の一実施形態に係る斜めに溝が設けられた反応槽の概略構成を示す。It is a perspective view which shows the variation of the reaction tank which concerns on this invention, (a) of FIG. 10 shows schematic structure of the semi-columnar reaction tank which concerns on one Embodiment of this invention, (b) of FIG. FIG. 10 shows a schematic configuration of a cut-out polygonal wall-like reaction tank according to an embodiment of the present invention, and FIG. 10C shows a reaction with oblique grooves according to an embodiment of the present invention. The schematic structure of a tank is shown. 本発明のいくつかの実施形態に係る溝または突起が設けられた反応層の概略構成を示す上面図であり、図１１の（ａ）は、本発明の一実施形態に係る片側に斜めに溝が設けられた反応槽の概略構成を示し、図１１の（ｂ）は、本発明の一実施形態に係る両側に斜めに溝が設けられた反応槽の概略構成を示す。FIG. 12 is a top view illustrating a schematic configuration of a reaction layer provided with grooves or protrusions according to some embodiments of the present invention, and FIG. 11A illustrates a groove obliquely on one side according to an embodiment of the present invention. FIG. 11B shows a schematic configuration of a reaction tank in which grooves are obliquely provided on both sides according to an embodiment of the present invention. 本発明の一実施形態に係るサンプル反応装置の概略構成を示す模式図であり、図１２の（ａ）は、本発明の一実施形態に係る、複数の反応槽を備えたサンプル反応装置の概略構成を示す断面図を示し、図１２の（ｂ）は、上記サンプル反応装置の概略構成を示す上面図を示す。It is a schematic diagram which shows schematic structure of the sample reaction apparatus which concerns on one Embodiment of this invention, (a) of FIG. 12 is the outline of the sample reaction apparatus provided with the several reaction tank based on one Embodiment of this invention. FIG. 12B is a top view showing a schematic configuration of the sample reaction apparatus. 本発明の一実施形態に係るサンプル反応装置を示す写真である。It is a photograph which shows the sample reaction apparatus which concerns on one Embodiment of this invention. 本発明に係るサンプル反応装置または従来技術を用いてサンプルを反応液と接触させた結果を示す写真であり、図１４の（ａ）～（ｃ）は、本発明に係るサンプル反応装置を用いてサンプルを反応液と接触させた結果を示し、図１４の（ｄ）は、従来技術を用いてサンプルを反応液と反応させた結果を示す。FIG. 15 is a photograph showing a result of contacting a sample with a reaction solution using a sample reaction apparatus according to the present invention or a conventional technique, and FIGS. 14 (a) to 14 (c) are drawings using the sample reaction apparatus according to the present invention. The result of contacting the sample with the reaction solution is shown. FIG. 14D shows the result of reacting the sample with the reaction solution using the conventional technique.
　本発明は、サンプル反応装置を提供する。本明細書において、サンプル反応装置とは、サンプルを反応液と接触させるための装置が意図される。 The present invention provides a sample reaction apparatus. In this specification, the sample reaction apparatus is intended to be an apparatus for bringing a sample into contact with a reaction solution.
　用語「サンプル」は当該分野において標本、調製物と同義で用いられ、本明細書中で使用される場合、「生物学的サンプル」またはその等価物が意図される。「生物学的サンプル」は、供給源としての生物材料（例えば、個体、体液、細胞株、組織培養物もしくは組織切片）から得られる、任意の調製物が意図される。 The term “sample” is used interchangeably with specimens, preparations in the art, and as used herein, “biological sample” or equivalent thereof is intended. A “biological sample” is intended to be any preparation obtained from biological material as a source (eg, an individual, body fluid, cell line, tissue culture or tissue section).
　生物学的サンプルとしては、体液（例えば、血液、唾液、歯垢、血清、血漿、尿、滑液、および随液）および組織供給源が挙げられる。好ましい生物学的サンプルは、被験体サンプルである。好ましい被験体サンプルは、被験体から得た皮膚病変部、喀痰、咽頭粘液、鼻腔粘液、膿、または分泌物である。本明細書中で使用される場合、用語「組織サンプル」は、組織供給源より得られた生物学的サンプルが意図される。哺乳動物から組織生検および体液を得るための方法は当該分野で周知である。 Biological samples include body fluids (eg, blood, saliva, plaque, serum, plasma, urine, synovial fluid, and fluids) and tissue sources. A preferred biological sample is a subject sample. Preferred subject samples are skin lesions, sputum, pharyngeal mucus, nasal mucus, pus, or secretions obtained from the subject. As used herein, the term “tissue sample” intends a biological sample obtained from a tissue source. Methods for obtaining tissue biopsies and body fluids from mammals are well known in the art.
　本明細書中で使用される場合、用語「サンプル」としては、上記生物学的サンプルおよび上記組織サンプル以外に、上記生物学的サンプルおよび上記組織サンプルより抽出したタンパク質サンプル、ゲノムＤＮＡサンプルおよび／または総ＲＮＡサンプルも挙げられる。 As used herein, the term “sample” includes, in addition to the biological sample and the tissue sample, a protein sample, genomic DNA sample and / or extracted from the biological sample and the tissue sample. A total RNA sample is also included.
　本明細書において、用語「反応液」は、サンプルと反応させるための液状の試薬が意図される。本発明に係るサンプル反応装置に用いるための反応液としては、例えば、抗体溶液、酵素溶液、染色液、洗浄液、緩衝液等を用いることができるが、特に限定されず、用いるサンプルおよび目的に応じて当該分野において周知慣用の試薬を用いることができる。 In the present specification, the term “reaction solution” is intended to be a liquid reagent for reacting with a sample. As a reaction solution for use in the sample reaction apparatus according to the present invention, for example, an antibody solution, an enzyme solution, a staining solution, a washing solution, a buffer solution, and the like can be used, but it is not particularly limited, depending on the sample to be used and the purpose. Conventional reagents well known in the art can be used.
　一例を挙げれば、本発明に係るサンプル反応装置を、ウェスタンブロッティング法により転写膜に転写されたタンパク質を抗体によって検出するために用いる場合、反応液としては、ブロッキング緩衝液、上記タンパク質と特異的に結合する一次抗体、該一次抗体と特異的に結合する二次抗体、洗浄液等のいずれかを好適に用いることができる。 For example, when the sample reaction apparatus according to the present invention is used for detecting a protein transferred to a transfer membrane by a Western blotting method using an antibody, the reaction solution is specifically a blocking buffer, and the above protein. Any of a primary antibody that binds, a secondary antibody that specifically binds to the primary antibody, a washing solution, and the like can be suitably used.
　また、本発明に係るサンプル反応装置を、サザンブロッティング法により転写膜に転写されたＤＮＡをプローブＤＮＡによって検出するために用いる場合、プローブＤＮＡを含んだ溶液を反応液として用いることができる。 Further, when the sample reaction apparatus according to the present invention is used to detect DNA transferred to the transfer film by the Southern blotting method using the probe DNA, a solution containing the probe DNA can be used as a reaction solution.
　さらに、反応液として、染色液を用いることにより、本発明に係るサンプル反応装置を、タンパク質の全自動染色装置として利用することもできる。上記染色液としては、タンパク質そのものを染色するもの、タンパク質の糖残基、リン酸基、ニトロ基等を染色するもの等を用いることができる。 Furthermore, by using a staining solution as a reaction solution, the sample reaction apparatus according to the present invention can be used as a fully automatic protein staining apparatus. As the staining solution, one that stains a protein itself, one that stains a sugar residue, a phosphate group, a nitro group, or the like of a protein can be used.
　そのような染色液としては、これに限定されるものではないが、市販の、SYPRO Ruby（インビトロジェン株式会社）、MemCode Reversible Protein Stain Kit（タカラバイオ株式会社）、GelCode Glycoprotein Staining Kit（テクノケミカル（株））、Deep Purple Total Protein Stain（GEヘルスケア）等を用いることができる。 Such staining solutions include, but are not limited to, commercially available SYPRO Ruby (Invitrogen Corporation), MemCode Reversible Protein Stain Kit (TAKARA BIO INC.), GelCode Glycoprotein Staining Kit (Technochemical Co., Ltd.) )), Deep, Purple, Total, Protein, and Stain (GE Healthcare).
　図１は、本発明の一実施形態に係るサンプル反応装置１００の概略構成を示す断面図である。また、図２は、サンプル反応装置１００の概略構成を示す斜視図である。図１および２に示すように、本実施形態に係るサンプル反応装置１００は、吸着構造体１１０と、反応槽１２０と、回転駆動部（駆動手段）１３０とを備えている。 FIG. 1 is a cross-sectional view showing a schematic configuration of a sample reaction apparatus 100 according to an embodiment of the present invention. FIG. 2 is a perspective view showing a schematic configuration of the sample reaction apparatus 100. As shown in FIGS. 1 and 2, the sample reaction apparatus 100 according to this embodiment includes an adsorption structure 110, a reaction tank 120, and a rotation drive unit (drive means) 130.
　吸着構造体１１０は、転写膜１１５と、転写膜支持体１１４とを備えている。転写膜１１５上には、サンプル１１１を吸着するサンプル吸着領域１１２が設けられている。 The adsorption structure 110 includes a transfer film 115 and a transfer film support 114. A sample adsorption region 112 that adsorbs the sample 111 is provided on the transfer film 115.
　また、反応槽１２０には、反応液１２１が貯められている。 Further, the reaction liquid 121 is stored in the reaction tank 120.
　回転駆動部１３０は、反応槽１２０側に設けられたモーター１３２およびギアセット１３３と、吸着構造体１１０に結合しているギア１３４とを備えている。 The rotation drive unit 130 includes a motor 132 and a gear set 133 provided on the reaction tank 120 side, and a gear 134 coupled to the adsorption structure 110.
　このように、サンプル反応装置１００は、サンプル１１１を吸着するサンプル吸着領域１１２を備えており、サンプル吸着領域１１２に吸着されたサンプル１１１と、反応液１２１とを接触させる。後述するように、本発明によれば、上記サンプル吸着領域の全域に渡って均一に、上記サンプルと上記反応液とを接触させることができる。そのため、本発明によれば、従来なし得なかった、サンプルを反応液とを均一に反応させることが可能である。 Thus, the sample reaction apparatus 100 includes the sample adsorption region 112 that adsorbs the sample 111, and the sample 111 adsorbed on the sample adsorption region 112 and the reaction solution 121 are brought into contact with each other. As will be described later, according to the present invention, the sample and the reaction liquid can be contacted uniformly over the entire area of the sample adsorption region. Therefore, according to the present invention, it is possible to uniformly react a sample with a reaction solution, which could not be achieved conventionally.
　サンプル吸着領域１１２は、サンプル１１１を吸着するものであればよく、サンプル１１１の吸着には、化学的、電気的、または物理的な特性等を用いることができる。例えば、当業者間において周知慣用されているように、転写膜１１５として、ナイロンメンブレン、ニトロセルロースメンブレン、ＰＶＤＦメンブレン等を用いる場合には、転写膜１１５は、その極性や、疎水性あるいはポアサイズによって、タンパク質やＤＮＡ等のサンプルを吸着させる。また、さらにベーキングや、ＵＶクロスリンク等の処理を行って、転写膜１１５とサンプル１１１とを強固に結合させてもよい。 The sample adsorption region 112 may be any as long as it adsorbs the sample 111, and chemical, electrical, physical characteristics, or the like can be used for adsorption of the sample 111. For example, as is well-known and commonly used by those skilled in the art, when a nylon membrane, a nitrocellulose membrane, a PVDF membrane, or the like is used as the transfer film 115, the transfer film 115 is determined depending on its polarity, hydrophobicity, or pore size. A sample such as protein or DNA is adsorbed. Furthermore, the transfer film 115 and the sample 111 may be firmly bonded by performing a process such as baking or UV cross-linking.
　本実施形態では、吸着構造体１１０は、転写膜１１５と、転写膜１１５を支持する転写膜支持体１１４とから構成されており、転写膜１１５上に、サンプル吸着領域１１２が設けられている。なお、吸着構造体１１０は、本実施形態のように、転写膜支持体１１４と、転写膜１１５とから構成されていてもよいが、例えば、吸着構造体１１０が、棒状、またはパイプ状であって、表面にサンプル吸着領域１１２が設けられている構成であってもよい。 In the present embodiment, the adsorption structure 110 includes a transfer film 115 and a transfer film support 114 that supports the transfer film 115, and a sample adsorption region 112 is provided on the transfer film 115. The adsorption structure 110 may be composed of a transfer film support 114 and a transfer film 115 as in this embodiment. For example, the adsorption structure 110 has a rod shape or a pipe shape. In addition, a configuration in which the sample adsorption region 112 is provided on the surface may be used.
　転写膜１１５上にサンプルを吸着させる方法は、特に限定されず、周知慣用の方法（例えば、ウェスタンブロッティング、細胞の溶解物等を滴下等）を用いることができる。また、吸着構造体１１０が棒状、またはパイプ状であって、表面にサンプル吸着領域１１２が設けられている構成である場合には、吸着構造体１１０を、サンプル１１１が吸着しているゲル等の上を転がすことにより、吸着構造体１１０上にサンプル１１１を吸着させてもよい。 The method for adsorbing the sample onto the transfer film 115 is not particularly limited, and a well-known and commonly used method (for example, western blotting, dropping of cell lysate, etc.) can be used. Further, when the adsorption structure 110 has a rod shape or a pipe shape and the sample adsorption region 112 is provided on the surface, the adsorption structure 110 is made of a gel or the like on which the sample 111 is adsorbed. The sample 111 may be adsorbed on the adsorption structure 110 by rolling up.
　反応槽１２０は、反応液１２１を貯め、サンプル吸着領域１１２が反応液１２１と接するように吸着構造体１１０を収納する。なお、サンプル１１１または反応液１２１の性質に応じて、反応槽１２０の底面にペルチェ素子などの温度調節部（温度調節手段）１２３を配置して、温度調節部１２３により反応液１２１の温度コントロールをおこなってもよい。 The reaction tank 120 stores the reaction solution 121 and stores the adsorption structure 110 so that the sample adsorption region 112 is in contact with the reaction solution 121. Depending on the properties of the sample 111 or the reaction liquid 121, a temperature adjustment unit (temperature adjustment means) 123 such as a Peltier element is disposed on the bottom surface of the reaction tank 120, and the temperature adjustment unit 123 controls the temperature of the reaction liquid 121. You may do it.
　回転駆動部（駆動手段）１３０は、吸着構造体１１０を、反応槽１２０内において、回転軸１３１の周りに回転させるものであればよい。本実施形態では、モーター１３２の回転がギアセット１３３に伝えられ、ギアセット１３３とギア１３４とがかみ合うことによって、吸着構造体１１０を回転させることができる。このとき、回転軸１３１は、反応槽１２０内に設定されている。ギアセット１３３は、ギア１３４の中心が回転軸１３１に一致する位置で、ギアセット１３３とギア１３４とがかみ合うように設けられている。 The rotation drive unit (drive means) 130 may be anything that rotates the adsorption structure 110 around the rotation axis 131 in the reaction tank 120. In the present embodiment, the rotation of the motor 132 is transmitted to the gear set 133, and the gear set 133 and the gear 134 are engaged with each other, whereby the adsorption structure 110 can be rotated. At this time, the rotating shaft 131 is set in the reaction tank 120. The gear set 133 is provided so that the gear set 133 and the gear 134 are engaged at a position where the center of the gear 134 coincides with the rotation shaft 131.
　なお、モーター１３２は、吸着構造体１１０の内部に設けられていてもよく、その場合は、ギア１３４が、処理槽１２０に固定されたギアセット１３３にかみ合うことによって、吸着構造体１１０を回転させる。 The motor 132 may be provided inside the adsorption structure 110. In that case, the gear 134 is engaged with a gear set 133 fixed to the processing tank 120, thereby rotating the adsorption structure 110. .
　ここで、図１および２に示すように、サンプル吸着領域１１２は、回転軸１３１を覆う面上に設けられている。それゆえ、吸着構造体１１０が回転することにより、サンプル吸着領域１１２と反応液１２１とを良好に接触させることができる。なお、回転軸１３１を覆う面とは、回転軸１３１を取り囲むように設けられた面の外面である。 Here, as shown in FIGS. 1 and 2, the sample adsorption region 112 is provided on a surface covering the rotating shaft 131. Therefore, when the adsorption structure 110 rotates, the sample adsorption region 112 and the reaction liquid 121 can be brought into good contact with each other. Note that the surface covering the rotation shaft 131 is an outer surface of a surface provided so as to surround the rotation shaft 131.
　図３は、転写膜支持体１１４の構造のバリエーションを示す斜視図である。転写膜支持体１１４は、図３の（ａ）に示すような棒状であってもよく、図３の（ｂ）に示すようなパイプ状の構造体であってもよく、図３の（ｃ）に示すようなスクリュー状の構造体であってもよく、図３の（ｄ）に示すようなメッシュ状の構造体であってもよい。 FIG. 3 is a perspective view showing a variation of the structure of the transfer film support 114. The transfer film support 114 may have a rod shape as shown in FIG. 3 (a) or a pipe-like structure as shown in FIG. 3 (b). A screw-like structure as shown in FIG. 3 or a mesh-like structure as shown in FIG.
　なお、転写膜支持体１１４を、図３の（ｃ）に示すようなスクリュー状の構造体、または、図３の（ｄ）に示すようなメッシュ状の構造体とすることにより、転写膜１１５との間に空隙を有するように転写膜１１５を支持することができる。転写膜支持体１１４が、転写膜１１５との間に空隙を有するように転写膜１１５を支持する場合、上記回転の際に、転写膜１１５の裏側にも反応液１２１が流動することになる。それゆえ、反応液１２１が滞留することがなく、サンプル１１１と、反応液１２１とをより好適に接触させることができる。 The transfer film support 114 is formed into a screw-like structure as shown in FIG. 3C or a mesh-like structure as shown in FIG. The transfer film 115 can be supported so as to have a gap between the two. When the transfer film support 114 supports the transfer film 115 so as to have a gap between the transfer film 115 and the transfer film 115, the reaction liquid 121 also flows on the back side of the transfer film 115 during the rotation. Therefore, the reaction liquid 121 does not stay, and the sample 111 and the reaction liquid 121 can be brought into contact more suitably.
　図４は、転写膜支持体１１４と転写膜１１５との結合様式のバリエーションを示す断面図である。転写膜１１５は、転写膜支持体１１４に設けられた結合部１１６に結合されている。結合部１１６の構造は、例えば、図４の（ａ）に示すような転写膜１１５を挟むスリットであってもよいし、図４の（ｂ）に示すような転写膜１１５を接着する接着物（磁石、両面テープ等）であってもよいし、図４の（ｃ）に示すような転写膜支持体１１４全体で転写膜１１５を挟み込む分割部であってもよく、転写膜１１５を支持することができる構造であれば特に限定されない。 FIG. 4 is a cross-sectional view showing variations in the coupling mode between the transfer film support 114 and the transfer film 115. The transfer film 115 is coupled to a coupling portion 116 provided on the transfer film support 114. The structure of the coupling portion 116 may be, for example, a slit that sandwiches the transfer film 115 as shown in FIG. 4A, or an adhesive that adheres the transfer film 115 as shown in FIG. 4B. (A magnet, a double-sided tape, or the like) or a divided portion that sandwiches the transfer film 115 with the entire transfer film support 114 as shown in FIG. 4C, and supports the transfer film 115. The structure is not particularly limited as long as it can be structured.
　図５～７は、サンプル吸着領域１１２が設けられる面１１３を説明する模式図である。面１１３は、回転軸１３１を覆うように設けられていればよいが、回転軸１３１と平行な面であることが好ましい。面１１３が、回転軸１３１と平行な面であれば、サンプル吸着領域１１２は、上記回転による移動の方向に直交する線から構成される面１１３上に設けられる。そのため、上記各線ごとにサンプル１１１の上記回転に伴う移動速度が等しくなる。よって、均一にサンプル１１１と反応液１２１とを接触させることができる。なお、特定の直線に平行な面とは、該直線とは交わらない面を指す。 5 to 7 are schematic diagrams for explaining the surface 113 on which the sample adsorption region 112 is provided. The surface 113 may be provided so as to cover the rotation shaft 131, but is preferably a surface parallel to the rotation shaft 131. If the surface 113 is a surface parallel to the rotation axis 131, the sample adsorption region 112 is provided on the surface 113 configured by a line orthogonal to the direction of movement by the rotation. Therefore, the moving speed accompanying the rotation of the sample 111 is equal for each line. Therefore, the sample 111 and the reaction liquid 121 can be contacted uniformly. Note that a plane parallel to a specific straight line refers to a plane that does not intersect the straight line.
　一つの局面において、面１１３は、図５の（ａ）に示すように、回転軸１３１を中心軸１１９とする円柱面である。この場合、図５の（ｂ）に示すように、面１１３上の各点は、反応液１２１内を同条件かつ滑らかに移動する。そのため、より均一にサンプル１１１と反応液１２１とを接触させることができる。 In one aspect, the surface 113 is a cylindrical surface having a rotation axis 131 as a central axis 119, as shown in FIG. In this case, as shown in FIG. 5B, each point on the surface 113 moves smoothly in the reaction solution 121 under the same conditions. Therefore, the sample 111 and the reaction liquid 121 can be contacted more uniformly.
　他の一つの局面において、面１１３は、図６の（ａ）に示すように、回転軸１３１に平行な軸を中心軸１１９とする円柱面である。この場合、図６の（ｂ）に示すように、面１１３上の各点は、反応液１２１内を滑らかに移動する。さらに、面１１３の回転により、反応液１２１が攪拌され、より好適にサンプル１１１と反応液１２１とを接触させることができる。 In another aspect, the surface 113 is a cylindrical surface having a central axis 119 as an axis parallel to the rotation axis 131 as shown in FIG. In this case, as shown in FIG. 6B, each point on the surface 113 moves smoothly in the reaction solution 121. Furthermore, the reaction liquid 121 is agitated by the rotation of the surface 113, and the sample 111 and the reaction liquid 121 can be brought into contact more suitably.
　なお、回転軸１３１と、面１１３の中心軸１１９との距離は、面１１３の半径の５０％以下であることが好ましい。上記距離が、上記半径の０％を超えれば、反応液１２１を攪拌することができる。上記距離が、上記半径の５０％以下であれば、反応液１２１とサンプル１１１とをより確実に接触させることができる。 Note that the distance between the rotation shaft 131 and the central axis 119 of the surface 113 is preferably 50% or less of the radius of the surface 113. When the distance exceeds 0% of the radius, the reaction liquid 121 can be stirred. When the distance is 50% or less of the radius, the reaction solution 121 and the sample 111 can be more reliably brought into contact with each other.
　さらに他の局面において、面１１３は、図７の（ａ）に示すように、回転軸１３１を中心軸１１９とする楕円柱面である。この場合、図７の（ｂ）に示すように、面１１３上の各点は、反応液１２１内を滑らかに移動する。さらに、面１１３の回転により、反応液１２１が攪拌され、より好適にサンプル１１１と反応液１２１とを接触させることができる。 In yet another aspect, the surface 113 is an elliptical cylinder surface having a rotation axis 131 as a central axis 119, as shown in FIG. In this case, as shown in FIG. 7B, each point on the surface 113 moves smoothly in the reaction solution 121. Furthermore, the reaction liquid 121 is agitated by the rotation of the surface 113, and the sample 111 and the reaction liquid 121 can be brought into contact more suitably.
　なお、面１１３の扁平率（長軸の長さ（ａ）－短軸の長さ（ｂ）／長軸の長さ（ａ））は、０．５以下であることが好ましい。上記扁平率が０を超えれば、反応液１２１を攪拌することができる。上記扁平率が０．５以下であれば、反応液１２１とサンプル１１１とをより均一に接触させることができる。 The flatness of the surface 113 (major axis length (a) −minor axis length (b) / major axis length (a)) is preferably 0.5 or less. When the flatness exceeds 0, the reaction liquid 121 can be stirred. When the flatness is 0.5 or less, the reaction solution 121 and the sample 111 can be contacted more uniformly.
　また、吸着構造体１１０は、サンプル吸着領域１１２を保護するための構造を備えていてもよい。図８は、カバー１１７を備えた吸着構造体１１０の概略構成を示す斜視図である。図９は、カバー１１７を備えた吸着構造体１１０の概略構成を示す断面図である。図８および９に示すように、カバー１１７は、転写膜１１５の外側に設けられ、サンプル吸着領域１１２を覆っている。そのため、サンプル吸着領域１１２に吸着したサンプル１１１が、他の物品と接触することを防ぐことができる。それゆえ、吸着構造体１１０を、サンプル１１１を吸着させたまま保存することが可能となる。 Further, the adsorption structure 110 may have a structure for protecting the sample adsorption region 112. FIG. 8 is a perspective view illustrating a schematic configuration of the adsorption structure 110 including the cover 117. FIG. 9 is a cross-sectional view illustrating a schematic configuration of the adsorption structure 110 including the cover 117. As shown in FIGS. 8 and 9, the cover 117 is provided outside the transfer film 115 and covers the sample adsorption region 112. Therefore, the sample 111 adsorbed on the sample adsorption region 112 can be prevented from coming into contact with other articles. Therefore, the adsorption structure 110 can be stored while the sample 111 is adsorbed.
　このような、カバー１１７を備えた吸着構造体１１０は、２ＤＥチップとして予め冷蔵保存しておき、病気になった時に、装置にセットすることによって病気の診断などに再利用することが可能である。すなわち、本発明はまた、上述したような構成を有するサンプル保存用チップを提供する。 Such an adsorption structure 110 provided with the cover 117 can be reused for diagnosing a disease by storing it in a refrigerated state as a 2DE chip in advance and setting it in a device when it becomes ill. . That is, the present invention also provides a sample storage chip having the above-described configuration.
　カバー１１７はまた、反応液１２１を透過させることが好ましい。そのようなカバー１１７としては、これに限定されるものではないが、開口部１１８を有し、開口部１１８から反応液１２１が入出可能であってもよい。開口部１１８は、図８に示すような切れ込み（すなわち、カバー１１７の一部が切り取られている）であってもよく、メッシュ状（すなわち、カバー１１７の一部がメッシュ状である）であってもよい。 It is preferable that the cover 117 also allows the reaction solution 121 to pass therethrough. Such a cover 117 is not limited to this, but may have an opening 118 so that the reaction liquid 121 can enter and exit from the opening 118. The opening 118 may be cut as shown in FIG. 8 (that is, a part of the cover 117 is cut out), and is mesh-shaped (that is, a part of the cover 117 is mesh-shaped). May be.
　このとき、図９に示すように、カバー１１７がセットされた状態で吸着構造体１１０を、反応槽１２０内に収納して回転させた場合であっても、開口部１１８を介して反応液１２１がサンプル吸着領域１１２に到達するため、サンプル１１１と、反応液１２１とを反応させることができる。なお、カバー１１７が、反応液１２１を透過しない場合は、吸着構造体１１０を、反応槽１２０に収納する前に、カバー１１７を除去すればよい。 At this time, as shown in FIG. 9, even when the adsorption structure 110 is housed in the reaction tank 120 and rotated with the cover 117 set, the reaction solution 121 is passed through the opening 118. Reaches the sample adsorption region 112, so that the sample 111 and the reaction liquid 121 can be reacted. If the cover 117 does not pass through the reaction solution 121, the cover 117 may be removed before the adsorption structure 110 is stored in the reaction tank 120.
　図１０は、反応槽１２０のバリエーションを示す斜視図である。図１０の（ａ）および（ｂ）に示すように、反応槽１２０の内面は、半円柱面、または半円柱面状、または六角柱以上の多角柱を長さ方向に対して平行な中心軸を通る面で切断したときの壁面状（但し、切断面は除く。すなわち、半円柱面に近似する形状）であることが好ましい。 FIG. 10 is a perspective view showing a variation of the reaction tank 120. As shown in (a) and (b) of FIG. 10, the inner surface of the reaction tank 120 is a semi-cylindrical surface, a semi-cylindrical surface shape, or a central axis parallel to the length direction of a hexagonal column or more polygonal column. It is preferable that the shape is a wall surface when cut by a plane passing through (excluding the cut surface, ie, a shape approximating a semi-cylindrical surface).
　反応槽１２０の内面が半円柱面であれば、該半円柱面の中心軸と、吸着構造体１１０の回転軸を同じ位置とすることにより、吸着構造体１１０を好適に収納することができる。特に、吸着構造体１１０の直径と、反応槽１２０の内面の直径がほぼ等しい場合、反応槽１２０内に、吸着構造体１１０をわずかな隙間で収納することができる。このわずかな隙間に反応液１２１を充填することにより、わずかな液量で、反応液１２１を、吸着構造体１１０の広い範囲に接触させることができ、反応液１２１の必要量を低減させることができる。なお、反応槽１２０の内面が、半円柱面ではなく、上記壁面状であっても、半円柱面に近い効果を得ることができる。 If the inner surface of the reaction tank 120 is a semi-cylindrical surface, the adsorption structure 110 can be suitably accommodated by setting the central axis of the semi-cylindrical surface and the rotation axis of the adsorption structure 110 to the same position. In particular, when the diameter of the adsorption structure 110 and the diameter of the inner surface of the reaction tank 120 are substantially equal, the adsorption structure 110 can be accommodated in the reaction tank 120 with a slight gap. By filling the reaction liquid 121 in this slight gap, the reaction liquid 121 can be brought into contact with a wide range of the adsorption structure 110 with a small liquid volume, and the required amount of the reaction liquid 121 can be reduced. it can. In addition, even if the inner surface of the reaction tank 120 is not a semi-cylindrical surface but the above wall surface shape, an effect close to the semi-cylindrical surface can be obtained.
　反応槽１２０の内面にはまた、図１０の（ｃ）に示すように回転軸１３１に平行な直線に斜めに交わる方向に沿った溝または突起が設けられていることが好ましい。図１１は、そのような溝または突起が設けられた反応槽１２０の上面図である。図１１の（ａ）は、反応槽１２０の片側に溝または突起が設けられた構成を示し、図１１の（ｂ）は、反応槽１２０の両側に溝または突起が設けられた構成を示す。 It is preferable that the inner surface of the reaction tank 120 is also provided with grooves or protrusions along a direction obliquely intersecting with a straight line parallel to the rotation shaft 131 as shown in FIG. FIG. 11 is a top view of the reaction vessel 120 provided with such grooves or protrusions. FIG. 11A shows a configuration in which grooves or protrusions are provided on one side of the reaction tank 120, and FIG. 11B shows a configuration in which grooves or protrusions are provided on both sides of the reaction tank 120.
　このとき、吸着構造体１１０の回転によって、反応液１２１が、該回転の方向に沿って流動するが、その流動方向は、上記の溝または突起に沿って斜めに方向転換され得る（図１１中矢印の方向）。それゆえ、反応液１２１の流動方向が一方向にならずに、反応液１２１が攪拌されるため、サンプル１１１と反応液１２１とをより均一に接触させることができる。 At this time, the reaction liquid 121 flows along the direction of the rotation due to the rotation of the adsorption structure 110, but the direction of the flow can be changed obliquely along the grooves or protrusions (in FIG. 11). Arrow direction). Therefore, since the reaction liquid 121 is stirred without the flow direction of the reaction liquid 121 being unidirectional, the sample 111 and the reaction liquid 121 can be more uniformly brought into contact with each other.
　本実施形態に係るサンプル反応装置１００は、例えば、二次元電気泳動によって分離されたタンパク質が転写された転写膜１１５について使用することができる。そのような転写膜は、面積が大きいため、従来技術では、全体を均一に反応液１２１と反応させることが困難であった。また、二次元電気泳動の解析のためには、特に、各部の濃度が均一であることが重要であるため、本実施形態に係るサンプル反応装置１００を特に好適に用いることができる。 The sample reaction apparatus 100 according to the present embodiment can be used for the transfer film 115 to which the protein separated by two-dimensional electrophoresis is transferred, for example. Since such a transfer film has a large area, it has been difficult for the transfer film to uniformly react with the reaction liquid 121 by the conventional technique. For the analysis of two-dimensional electrophoresis, it is particularly important that the concentration of each part is uniform, so that the sample reaction apparatus 100 according to the present embodiment can be used particularly suitably.
　本発明に係るサンプル反応装置はまた、反応槽を複数備えている構成であってもよい。図１２は、本発明の他の実施形態に係るサンプル反応装置２００の概略構成を示す模式図であり、図１２の（ａ）は断面図を示し、図１２の（ｂ）は上面図を示す。 The sample reaction apparatus according to the present invention may be configured to include a plurality of reaction vessels. FIG. 12 is a schematic diagram showing a schematic configuration of a sample reaction apparatus 200 according to another embodiment of the present invention, in which (a) of FIG. 12 shows a sectional view and (b) of FIG. 12 shows a top view. .
　図１２に示すように、本実施形態に係るサンプル反応装置２００は、吸着構造体２１０と、反応槽２２０、２２２、２２４、および２２６と、廃液槽２２８と、駆動部（駆動手段）２３０とを備えている。反応槽２２０、２２２、２２４、および２２６には、それぞれ反応液２２１、２２３、２２５、および２２７が貯められている。駆動部２３０は、モーター２３２、ギアセット２３３およびアーム２３４を備えている。 As shown in FIG. 12, the sample reaction apparatus 200 according to this embodiment includes an adsorption structure 210, reaction tanks 220, 222, 224, and 226, a waste liquid tank 228, and a drive unit (drive means) 230. I have. Reaction liquids 221, 223, 225, and 227 are stored in the reaction tanks 220, 222, 224, and 226, respectively. The drive unit 230 includes a motor 232, a gear set 233, and an arm 234.
　吸着構造体２１０としては、上述したサンプル反応装置１００における吸着構造体１１０と同様のものを用いることができる。アーム２３４は、吸着構造体２１０を、任意の反応槽に移動させる。アーム２３４は、例えば、周知慣用のロボットアーム技術を用いて構成すればよい。 As the adsorption structure 210, the same one as the adsorption structure 110 in the sample reaction apparatus 100 described above can be used. The arm 234 moves the adsorption structure 210 to an arbitrary reaction tank. The arm 234 may be configured using, for example, a well-known and common robot arm technique.
　したがって、本実施形態に係るサンプル反応装置２００を用いれば、所定の反応槽に吸着構造体２１０を移動させることにより、一連の反応を順次行うことができる。例えば、サンプル反応装置２００を、（ｉ）ブロッキング処理、（ｉｉ）一次抗体との反応、（ｉｉｉ）洗浄、および（ｉｖ）二次抗体との反応の一連の反応に用いることができる。この場合、反応液２２１としては、ブロッキング緩衝液、反応液２２３としては、一次抗体を含んだブロッキング緩衝液、反応液２２５としては、二次抗体を含んだブロッキング反応液、反応液２２７としては洗浄液とすることができる。 Therefore, by using the sample reaction apparatus 200 according to this embodiment, a series of reactions can be performed sequentially by moving the adsorption structure 210 to a predetermined reaction tank. For example, the sample reaction apparatus 200 can be used for a series of reactions of (i) blocking treatment, (ii) reaction with a primary antibody, (iii) washing, and (iv) reaction with a secondary antibody. In this case, the reaction solution 221 is a blocking buffer solution, the reaction solution 223 is a blocking buffer solution containing a primary antibody, the reaction solution 225 is a blocking reaction solution containing a secondary antibody, and the reaction solution 227 is a washing solution. It can be.
　また、反応槽２２６には、反応液２２７としての洗浄液を、例えば、被添加部２２９に対して、逐次添加してもよい。その場合、あふれ出た反応液２２７を回収する廃液槽２２８を、図４に示すように、反応槽２２６に隣接させて設ければよい。反応液２２７の添加手段２３０としては、例えば、当業者間において周知慣用されているバルブ付の管等を用いることができる。 In addition, a cleaning liquid as the reaction liquid 227 may be sequentially added to the reaction tank 226, for example, to the part to be added 229. In that case, a waste liquid tank 228 for collecting the overflowing reaction liquid 227 may be provided adjacent to the reaction tank 226 as shown in FIG. As the addition means 230 of the reaction liquid 227, for example, a tube with a valve well known and commonly used by those skilled in the art can be used.
　上述したように、本実施形態に係るサンプル反応装置２００を用いれば、一連の反応を順次行うことができるため、ウェスタンブロッティング結果の検出等の煩雑な工程を自動化することができる。サンプル反応装置２００の一実施例の写真を図１３に示す。 As described above, if the sample reaction apparatus 200 according to this embodiment is used, a series of reactions can be performed sequentially, so that complicated processes such as detection of Western blotting results can be automated. A photograph of an example of the sample reactor 200 is shown in FIG.
　すなわち、他の観点から言えば、サンプル反応装置２００は、複数の反応槽を備え、サンプルを吸着する吸着構造体を、各反応槽を順次移動させることにより、一連の反応を自動化できることを目的の１つとしている。 That is, from another point of view, the sample reaction apparatus 200 includes a plurality of reaction vessels, and an object of the present invention is to automate a series of reactions by sequentially moving an adsorption structure that adsorbs a sample in each reaction vessel. One.
　本発明は上述した各実施形態に限定されるものではなく、請求項に示した範囲で種々の変更が可能であり、異なる実施形態にそれぞれ開示された技術的手段を適宜組み合わせて得られる実施形態についても本発明の技術的範囲に含まれる。 The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope shown in the claims, and embodiments obtained by appropriately combining technical means disclosed in different embodiments. Is also included in the technical scope of the present invention.
　また、本明細書中に記載された学術文献および特許文献の全てが、本明細書中において参考として援用される。 In addition, all the academic literatures and patent literatures described in this specification are incorporated herein by reference.
　（サンプルが吸着された転写膜の調製）
　転写膜としては、ＰＶＤＦ膜（Ｉｍｍｏｂｉｒｏｎ－ｐＳＱ、Ｍｉｌｌｉｐｏｒｅ）を用いた。サンプルとしては、Ｃａｒｂｏｎｉｃ　ａｎｈｙｄｒａｓｅ　ｆｒｏｍ　Ｂｏｖｉｎｅ　Ｅｒｙｔｈｒｏｃｙｔｅｓ（Ｃ－３９３４、Ｓｉｇｍａ　Ｃｏ．）の希釈系列（＃１：５．６３ｎｍｏｌｅ、＃２：５６３ｐｍｏｌｅ、＃３：５６．３ｐｍｏｌｅ、＃４：５．６３ｐｍｏｌｅ、＃５：５６３ｆｍｏｌｅ、＃６：５６．３ｆｍｏｌｅ）を用いた。希釈には蒸留水を用いた。 (Preparation of transfer film with sample adsorbed)
As the transfer film, a PVDF film (Immobilon-pSQ, Millipore) was used. As a sample, dilution series (# 1: 5.63 nmole, # 2: 563 pmole, # 3: 56.3 pmole, # 4: 5.63 pmole, # 5 of Carbon anhydrase from Bovine Erythrocytes (C-3934, Sigma Co.) : 563 fmol, # 6: 56.3 fmol). Distilled water was used for dilution.
　上記転写膜を、スクリーナーブロッター（サンプラテック社）に設置し、上記サンプルをそれぞれ８５μｌずつアプライし、４０分間静置して、該転写膜上に該サンプルを吸着させた。 The transfer film was placed on a screener blotter (Samplatech), 85 μl of each sample was applied, and allowed to stand for 40 minutes to adsorb the sample onto the transfer film.
　（ブロッキング工程）
　次に、上記転写膜をブロッキング緩衝液（１．５％スキムミルクを含むＴＢＳ－Ｔ（０．０５％Ｔｗｅｅｎ２０、５０ｍＭ　Ｔｒｉｓ（２４．２ｇ／４Ｌ）、１５０ｍＭ　ＮａＣｌ（３５ｇ／４Ｌ）））と３０分間反応させた。 (Blocking process)
Next, the transfer membrane is treated with a blocking buffer (TBS-T containing 1.5% skim milk (0.05% Tween 20, 50 mM Tris (24.2 g / 4 L), 150 mM NaCl (35 g / 4 L))) for 30 minutes. Reacted.
　（一次抗体との反応工程）
　ブロッキング処理後、上記転写膜を、ブロッキング緩衝液で希釈した一次抗体（抗Ｃａｒｂｏｎｉｃ　ａｎｈｙｄｒａｓｅ　ｆｒｏｍ　Ｂｏｖｉｎｅ　Ｅｒｙｔｈｒｏｃｙｔｅｓ　ウサギ抗体（ＮＥ０２３／７Ｓ、Ｎｏｒｄｉｃ　Ｉｍｍｕｎｏｌｏｇｉｃａｌ　Ｌａｂ．））と反応させた。 (Reaction process with primary antibody)
After the blocking treatment, the transfer membrane was reacted with a primary antibody diluted with a blocking buffer (anti-carbonic anhydrobrom from Bovine Erythrocytes rabbit antibody (NE023 / 7S, Nordic Immunological Lab.)).
　（洗浄工程－１）
　一次抗体との反応後、上記転写膜を、洗浄液（ＴＢＳ－Ｔ）により洗浄した。洗浄は３回５分ずつ行った。 (Washing process-1)
After the reaction with the primary antibody, the transfer film was washed with a washing solution (TBS-T). Washing was performed three times for 5 minutes each.
　（二次抗体との反応工程）
　洗浄後、上記転写膜を、ブロッキング緩衝液で希釈した二次抗体（Ａｌｅｘａ　Ｆｌｕｏｒ　６４７　ｇｏａｔ　ａｎｔｉ－ｒａｂｂｉｔ　ＩｇＧ（Ｈ＋Ｌ）（Ａ２１２４４、Ｉｎｖｉｔｒｏｇｅｎ　Ｃｏ．））と反応させた。 (Reaction process with secondary antibody)
After washing, the transfer membrane was reacted with a secondary antibody (Alexa Fluor 647 goat anti-rabbit IgG (H + L) (A21244, Invitrogen Co.)) diluted with a blocking buffer.
　（洗浄工程－２）
　一次抗体との反応後、上記転写膜を、洗浄液（ＴＢＳ－Ｔ）により５分ずつ３回洗浄した。その後さらに、洗浄液（ＴＢＳ（５０ｍＭ　Ｔｒｉｓ（２４．２ｇ／４Ｌ）、１５０ｍＭ　ＮａＣｌ（３５ｇ／４Ｌ））により５分間洗浄した。 (Washing process-2)
After the reaction with the primary antibody, the transfer film was washed with a washing solution (TBS-T) three times for 5 minutes each. Thereafter, it was further washed with a washing solution (TBS (50 mM Tris (24.2 g / 4 L), 150 mM NaCl (35 g / 4 L)) for 5 minutes.
　得られた転写膜上のバンドを、蛍光スキャナー（Ｔｙｐｈｏｏｎ　ＴＲＩＯ＋、ＧＥヘルスケア社）により検出した。 The band on the obtained transfer film was detected by a fluorescence scanner (Typhoon TRIO +, GE Healthcare).
　〔実施例１〕
　上述した、ブロッキング処理、一次抗体との反応、洗浄、および二次抗体との反応を、図１２に示すような、ブロッキング処理、一次抗体との反応、洗浄、および二次抗体との反応の、それぞれの反応のための反応槽を備えた、本発明に係るサンプル反応装置を用いて行った。また、転写膜支持体として、直径１９ｍｍの、図３の（ｃ）に示すようなスクリュー形状のものを用いた。また、ブロッキング処理、一次抗体との反応、および二次抗体との反応のための反応槽として、直径２０ｍｍの半円柱形状のものを用い、洗浄のための反応槽として、直径２２ｍｍの半円柱形状のものを用いた。すべての反応は、２０℃、湿度７０％の条件下で行った。 [Example 1]
The blocking treatment, the reaction with the primary antibody, the washing, and the reaction with the secondary antibody described above are shown in FIG. 12 in the blocking treatment, the reaction with the primary antibody, the washing, and the reaction with the secondary antibody. The reaction was carried out using a sample reaction apparatus according to the present invention equipped with a reaction tank for each reaction. A transfer film support having a diameter of 19 mm and having a screw shape as shown in FIG. 3C was used. In addition, as a reaction vessel for blocking treatment, reaction with the primary antibody, and reaction with the secondary antibody, a semi-cylindrical shape having a diameter of 20 mm is used, and as a reaction vessel for washing, a semi-cylindrical shape having a diameter of 22 mm is used. The thing of was used. All reactions were performed under conditions of 20 ° C. and 70% humidity.
　ブロッキング工程、一次抗体との反応工程、洗浄工程、二次抗体との反応工程は、それぞれのための反応槽内において、上記転写膜を支持する支持体を１８回転／分の速度で回転させることにより行った。 In the blocking step, the reaction step with the primary antibody, the washing step, and the reaction step with the secondary antibody, the support for supporting the transfer film is rotated at a speed of 18 revolutions / minute in the reaction tank for each. It went by.
　ブロッキング工程における、ブロッキング緩衝液の液量は５ｍｌとした。一次抗体または二次抗体との反応工程における反応液は、１μｌの上記抗体を全量５ｍｌとなるように希釈したものを用い、反応時間は６０分間とした。 The amount of blocking buffer in the blocking step was 5 ml. The reaction solution in the reaction step with the primary antibody or the secondary antibody was prepared by diluting 1 μl of the above antibody so that the total amount became 5 ml, and the reaction time was 60 minutes.
　結果を図１４の（ａ）に示す。 The results are shown in FIG.
　〔実施例２〕
　一次抗体または二次抗体との反応工程における反応時間を１５分間とした以外は、実施例１と同様に反応を行った。結果を図１４の（ｂ）に示す。 [Example 2]
The reaction was carried out in the same manner as in Example 1 except that the reaction time in the reaction step with the primary antibody or the secondary antibody was 15 minutes. The result is shown in FIG.
　〔実施例３〕
　一次抗体または二次抗体との反応工程における反応液を、１μｌの上記抗体を全量１．２３ｍｌとなるように希釈したものを用い、反応時間を１５分間とした以外は、実施例１と同様に反応を行った。結果を図１４の（ｃ）に示す。 Example 3
As in Example 1, except that the reaction solution in the reaction step with the primary antibody or the secondary antibody was prepared by diluting 1 μl of the above antibody to a total volume of 1.23 ml, and setting the reaction time to 15 minutes. Reaction was performed. The results are shown in FIG.
　〔比較例１〕
　上述した、ブロッキング処理、一次抗体との反応、洗浄、および二次抗体との反応を、従来法により行った。具体的には、袋中に、上記転写膜と、上記各反応液とを投入した後、該袋を密封して袋ごと１８回転／分の速度で回転させた。反応はすべて室温で行った。 [Comparative Example 1]
The above-described blocking treatment, reaction with the primary antibody, washing, and reaction with the secondary antibody were performed by conventional methods. Specifically, after the transfer film and each of the reaction liquids were put into a bag, the bag was sealed and rotated at a speed of 18 rotations / minute together with the bag. All reactions were performed at room temperature.
　ブロッキング工程における、ブロッキング緩衝液の液量は２５ｍｌとした。一次抗体または二次抗体との反応工程における反応液は、５μｌの上記抗体を全量５ｍｌとなるように希釈したものを用い、反応時間は６０分間とした。洗浄工程における洗浄液の液量は２５ｍｌとした。結果を図１４の（ｄ）に示す。 The amount of blocking buffer in the blocking step was 25 ml. The reaction solution in the reaction step with the primary antibody or the secondary antibody was prepared by diluting 5 μl of the above antibody so that the total amount became 5 ml, and the reaction time was 60 minutes. The amount of the cleaning liquid in the cleaning process was 25 ml. The result is shown in FIG.
　図１４の（ａ）～（ｃ）と図１４の（ｄ）とを比較すれば判るように、従来法により得られたバンドは、中央部が薄くなっている（図１４の（ｄ）参照）のに対し、本発明に係るサンプル反応装置を用いた場合は、全体が均一に発色している（図１４の（ａ）～（ｃ）参照）。 As can be seen by comparing (a) to (c) of FIG. 14 and (d) of FIG. 14, the band obtained by the conventional method has a thin central portion (see (d) of FIG. 14). In contrast, when the sample reaction apparatus according to the present invention is used, the whole color is uniformly developed (see FIGS. 14A to 14C).
　また、本発明に係るサンプル反応装置を用いることにより、従来法により得られたバンド（図１４の（ｄ）参照）とほぼ同等の発色が、一次抗体および二次抗体の濃度を薄くした場合（図１４の（ａ）参照）、該濃度を薄くし、さらに一次抗体および二次抗体との反応時間を短くした場合（図１４の（ｂ）参照）、ならびに、一次抗体および二次抗体との反応工程において、反応液の液量を少なくし、さらに反応時間を短くした場合（図１４の（ｃ）参照）においても得られた。 In addition, when the sample reaction apparatus according to the present invention is used, color development substantially equivalent to the band obtained by the conventional method (see (d) of FIG. 14) is obtained when the concentrations of the primary antibody and the secondary antibody are reduced ( 14 (a)), when the concentration is reduced and the reaction time with the primary antibody and the secondary antibody is shortened (see FIG. 14 (b)), and with the primary antibody and the secondary antibody In the reaction step, it was also obtained when the amount of the reaction solution was reduced and the reaction time was further shortened (see FIG. 14 (c)).
　本発明に係るサンプル反応装置は、サンプルを吸着するサンプル吸着領域を備えている吸着構造体を、反応液を貯める反応槽内で回転させるので、該サンプルを該反応液と均一に反応させることができる。 In the sample reaction apparatus according to the present invention, the adsorption structure having the sample adsorption region for adsorbing the sample is rotated in the reaction tank for storing the reaction solution, so that the sample can be reacted uniformly with the reaction solution. it can.
　発明の詳細な説明の項においてなされた具体的な実施形態または実施例は、あくまでも、本発明の技術内容を明らかにするものであって、そのような具体例にのみ限定して狭義に解釈されるべきものではなく、本発明の精神と次に記載する請求の範囲内で、いろいろと変更して実施することができるものである。 The specific embodiments or examples made in the detailed description section of the invention are merely to clarify the technical contents of the present invention, and are limited to such specific examples and are interpreted in a narrow sense. It should be understood that various modifications may be made within the spirit of the invention and the scope of the following claims.
　本発明は、各種実験装置、検査装置、および測定装置等の分野において利用可能である。 The present invention can be used in the fields of various experimental devices, inspection devices, measuring devices, and the like.
　１００　　　　　サンプル反応装置
　１１０　　　　　吸着構造体
　１１１　　　　　サンプル
　１１２　　　　　サンプル吸着領域
　１１３　　　　　面
　１１４　　　　　転写膜支持体
　１１５　　　　　転写膜
　１１６　　　　　結合部
　１１７　　　　　カバー
　１１８　　　　　開口部
　１１９　　　　　中心軸
　１２０　　　　　反応槽
　１２１　　　　　反応液
　１２２　　　　　溝
　１２３　　　　　温度調節部（温度調節手段）
　１３０　　　　　回転駆動部（駆動手段）
　１３１　　　　　回転軸
　１３２　　　　　モーター
　１３３　　　　　ギアセット
　１３４　　　　　ギア
　２００　　　　　サンプル反応装置
　２１０　　　　　吸着構造体
　２２０　　　　　第１反応槽
　２２１　　　　　第１反応液
　２２２　　　　　第２反応槽
　２２３　　　　　第２反応液
　２２４　　　　　第３反応槽
　２２５　　　　　第３反応液
　２２６　　　　　第４反応槽
　２２７　　　　　第５反応液
　２２８　　　　　廃液槽
　２２９　　　　　被添加部
　２３０　　　　　添加手段
　２３２　　　　　モーター
　２３３　　　　　ギアセット
　２３４　　　　　アーム DESCRIPTION OF SYMBOLS 100 Sample reaction apparatus 110 Adsorption structure 111 Sample 112 Sample adsorption area 113 Surface 114 Transfer film support body 115 Transfer film 116 Coupling part 117 Cover 118 Opening part 119 Central axis 120 Reaction tank 121 Reaction liquid 122 Groove 123 Temperature control part (Temperature control part) means)
130 Rotation drive part (drive means)
131 Rotating shaft 132 Motor 133 Gear set 134 Gear 200 Sample reaction device 210 Adsorption structure 220 First reaction tank 221 First reaction liquid 222 Second reaction tank 223 Second reaction liquid 224 Third reaction tank 225 Third reaction liquid 226 First 4 reaction tank 227 fifth reaction liquid 228 waste liquid tank 229 added part 230 addition means 232 motor 233 gear set 234 arm

Claims (20)

  1. 　サンプルを吸着するサンプル吸着領域を備えている吸着構造体と、
    　反応液を貯め、該サンプル吸着領域が該反応液と接するように該吸着構造体を収納する反応槽と、
    　該吸着構造体を該反応槽内で回転させる駆動手段とを備えており、
    　該サンプル吸着領域は、該回転の回転軸を覆う面上に設けられていることを特徴とするサンプル反応装置。     An adsorption structure having a sample adsorption area for adsorbing a sample;
    A reaction tank for storing the reaction solution and storing the adsorption structure so that the sample adsorption region is in contact with the reaction solution;
    Drive means for rotating the adsorption structure in the reaction vessel,
    The sample reaction apparatus, wherein the sample adsorption region is provided on a surface covering the rotation axis of the rotation.
  2. 　上記回転軸を覆う面が、上記回転軸と平行な面であることを特徴とする請求の範囲第１項に記載のサンプル反応装置。 The sample reaction apparatus according to claim 1, wherein a surface covering the rotation axis is a plane parallel to the rotation axis.
  3. 　上記回転軸を覆う面が、円柱面、または楕円柱面であることを特徴とする請求の範囲第１項または第２項に記載のサンプル反応装置。 3. The sample reaction apparatus according to claim 1, wherein the surface covering the rotation axis is a cylindrical surface or an elliptical cylindrical surface.
  4. 　上記吸着構造体が、上記サンプル吸着領域が設けられている転写膜と、該転写膜を支持する転写膜支持体とをさらに備えていることを特徴とする請求の範囲第１項～第３項のいずれか一項に記載のサンプル反応装置。 The first to third claims, wherein the adsorption structure further comprises a transfer film provided with the sample adsorption region, and a transfer film support for supporting the transfer film. The sample reactor according to any one of the above.
  5. 　上記転写膜支持体と上記転写膜との間に空隙が設けられていることを特徴とする請求の範囲第４項に記載のサンプル反応装置。 The sample reaction apparatus according to claim 4, wherein a gap is provided between the transfer film support and the transfer film.
  6. 　上記転写膜支持体に、上記回転軸に平行な直線に斜めに交わる方向に沿った溝が設けられていることを特徴とする請求の範囲第４項または第５項に記載のサンプル反応装置。 The sample reaction apparatus according to claim 4 or 5, wherein the transfer film support is provided with a groove along a direction obliquely intersecting a straight line parallel to the rotation axis.
  7. 　上記吸着構造体が、上記サンプル吸着領域を覆うカバーをさらに備えていることを特徴とする請求の範囲第１項～第６項の何れか一項に記載のサンプル反応装置。 The sample reaction apparatus according to any one of claims 1 to 6, wherein the adsorption structure further includes a cover that covers the sample adsorption region.
  8. 　上記カバーが、上記反応液を透過させることを特徴とする請求の範囲第７項に記載のサンプル反応装置。 The sample reaction apparatus according to claim 7, wherein the cover allows the reaction solution to pass therethrough.
  9. 　上記反応槽の内面が、半円柱面状、または六角柱以上の多角柱を長さ方向に対して平行な中心軸を通る面で切断したときの壁面状であることを特徴とする請求の範囲第１項～第８項のいずれか一項に記載のサンプル反応装置。 The inner surface of the reaction tank is a semi-cylindrical surface shape or a wall surface shape when a polygonal column of hexagonal column or higher is cut by a plane passing through a central axis parallel to the length direction. 9. The sample reaction apparatus according to any one of items 1 to 8.
  10. 　上記反応槽の内面に、上記回転軸に平行な直線に斜めに交わる方向に沿った溝または突起が設けられていることを特徴とする請求の範囲第１項～第９項のいずれか一項に記載のサンプル反応装置。 The groove or protrusion along a direction obliquely intersecting with a straight line parallel to the rotation axis is provided on the inner surface of the reaction vessel. The sample reactor according to 1.
  11. 　上記反応槽に上記反応液を添加する添加手段と、該反応槽から流出する該反応液を受ける廃液槽とをさらに備えていることを特徴とする請求の範囲第１項～第１０項のいずれか一項に記載のサンプル反応装置。 11. The apparatus according to claim 1, further comprising addition means for adding the reaction liquid to the reaction tank and a waste liquid tank for receiving the reaction liquid flowing out from the reaction tank. The sample reaction apparatus according to claim 1.
  12. 　複数の上記反応槽を備えており、上記駆動手段が、上記吸着構造体を任意の該反応槽に移動させることを特徴とする請求の範囲第１項～第１１項のいずれか一項に記載のサンプル反応装置。 12. The apparatus according to any one of claims 1 to 11, further comprising a plurality of the reaction vessels, wherein the driving unit moves the adsorption structure to an arbitrary reaction vessel. Sample reactor.
  13. 　上記サンプルがタンパク質であり、上記反応液が、該タンパク質の抗体を含んでいることを特徴とする請求の範囲第１項～第１２項のいずれか一項に記載のサンプル反応装置。 The sample reaction apparatus according to any one of claims 1 to 12, wherein the sample is a protein, and the reaction solution contains an antibody of the protein.
  14. 　上記サンプルがタンパク質であり、上記複数の反応槽が、ブロッキング緩衝液を貯める第１反応槽、該タンパク質に結合する一次抗体を含む反応液を貯める第２反応槽、該一次抗体に結合する二次抗体を含む反応液を貯める第３反応槽、および洗浄液を貯める第４反応槽を含んでいることを特徴とする請求の範囲第１２項に記載のサンプル反応装置。 The sample is a protein, and the plurality of reaction tanks store a first reaction tank that stores a blocking buffer, a second reaction tank that stores a reaction liquid containing a primary antibody that binds to the protein, and a secondary that binds to the primary antibody. 13. The sample reaction apparatus according to claim 12, further comprising a third reaction tank for storing a reaction liquid containing an antibody and a fourth reaction tank for storing a washing liquid.
  15. 　サンプルを吸着する転写膜を支持する転写膜支持体と、
    　反応液を貯め、該転写膜支持体が支持する転写膜が該反応液と接するように該転写膜支持体を収納する反応槽と、
    　該転写膜支持体を該反応槽内で回転させる駆動手段とを備えており、
    　該転写膜支持体は、該転写膜を、該回転の回転軸を覆う面上に支持することを特徴とするサンプル反応装置。     A transfer film support that supports the transfer film that adsorbs the sample;
    A reaction tank for storing the reaction liquid and storing the transfer film support so that the transfer film supported by the transfer film support is in contact with the reaction liquid;
    Drive means for rotating the transfer film support in the reaction vessel,
    The sample reaction apparatus, wherein the transfer film support supports the transfer film on a surface covering a rotation axis of the rotation.
  16. 　サンプルを吸着するサンプル吸着領域を備えている吸着構造体を、反応液が貯められた反応槽内で回転させる工程を包含しており、
    　該サンプル吸着領域は、該回転の回転軸を覆う面上に設けられていることを特徴とするサンプル反応方法。     Including a step of rotating an adsorption structure having a sample adsorption region for adsorbing a sample in a reaction tank in which a reaction solution is stored;
    The sample reaction method, wherein the sample adsorption region is provided on a surface covering a rotation axis of the rotation.
  17. 　一つの軸を覆うように設けられている、サンプルを吸着するサンプル吸着領域と、該サンプル吸着領域を覆うカバーとを備えていることを特徴とするサンプル保存用チップ。 A sample storage chip comprising a sample adsorption region for adsorbing a sample and a cover for covering the sample adsorption region, which are provided so as to cover one axis.
  18. 　サンプルを吸着するサンプル吸着領域を備えている吸着構造体と、
    　反応液を貯め、該サンプル吸着領域が該反応液と接するように該吸着構造体を収納する複数の反応槽と、
    　該吸着構造体を任意の該反応槽に移動させる駆動手段とを備えていることを特徴とするサンプル反応装置。     An adsorption structure having a sample adsorption area for adsorbing a sample;
    A plurality of reaction tanks for storing the reaction solution and storing the adsorption structure so that the sample adsorption region is in contact with the reaction solution;
    A sample reaction apparatus comprising: a drive unit that moves the adsorption structure to an arbitrary reaction tank.
  19. 　サンプルを吸着するサンプル吸着領域を備えている吸着構造体を、第１反応液が貯められた第１反応槽内に移動させる第１反応工程と、
    　第１反応工程の後に、上記吸着構造体を、第２反応液が貯められた第２反応槽に移動させる第２反応工程とを包含することを特徴とするサンプル反応方法。     A first reaction step of moving an adsorption structure having a sample adsorption region for adsorbing a sample into a first reaction tank in which a first reaction liquid is stored;
    A sample reaction method comprising a second reaction step of moving the adsorption structure to a second reaction tank in which a second reaction solution is stored after the first reaction step.
  20. 　上記反応槽が、上記反応液の温度を調節する温度調節手段を備えていることを特徴とする請求の範囲第１項～第１５項および第１８項のいずれか一項に記載のサンプル反応装置。 The sample reaction apparatus according to any one of claims 1 to 15, wherein the reaction tank includes a temperature adjusting means for adjusting the temperature of the reaction solution. .
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