Device, Chip, and Substrate

Abstract

A device (100) according to the present invention captures and senses a detected substance in a liquid specimen, the device comprising: a base material (101); a flow path (103) which is provided to one surface of the base material (101) and along which the liquid specimen is fed; and a sensing zone (110) provided to one section of the flow path (103). A gel-like pillar (105) constituted by a gel-like substance is provided to the sensing zone (110), a captured substance (107) which specifically binds to the detected substance is held in the gel-like pillar (105), and the flow path (103) is formed by a plurality of flow path forming pillars (109) or microgrooves upstream or downstream of the sensing zone (110).

Description

Technical Field

[0001] The present invention relates to devices, chips, and substrates.

Background Art

[0002] As technologies related to devices for detecting components in a sample, there are those described in Patent Documents 1 to 3. Patent Document 1 describes an immunoassay microchip provided with a flow path in which at least a part of a micro-structure in which fine beads having a primary antibody immobilized on the surface are homogeneously dispersed and held in a photocured hydrophilic resin is arranged.

[0003] Patent Document 2 describes a microfluidic device in which micro-structures in which a kind of specific binding reagent or a kind of sample mixed in a photocured hydrophilic resin is held by crosslinking are arranged in individual flow paths provided on a substrate.

[0004] Further, Patent Document 3 describes a diagnostic element including an inlet passage; a holding port embedding a diagnostic gel containing pores; and an outlet passage, wherein the inlet passage and the outlet passage are on either side of the holding port.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0006] The present invention provides a device that has excellent detection sensitivity and can be easily used even without using an additional device for liquid feeding.

Means for Solving the Problems

[0007] According to the present invention, the following devices, chips, and substrates are provided.

[0008] [1] A device for capturing and detecting a substance to be detected in a liquid sample, a base material, a flow path provided on one surface of the base material for transferring the liquid sample, a detection zone provided in a part of the flow path, and a gel-like pillar made of a gel-like substance is provided in the detection zone, and a capturing substance that specifically binds to the substance to be detected is held in the gel-like pillar, a device in which, on the upstream side or downstream side of the detection zone, the flow path is formed by a plurality of flow path forming pillars or fine grooves. [2] An induction zone for guiding the liquid sample to the detection zone by capillary action is provided on the upstream side of the detection zone, the device according to [1], wherein in the induction zone, the flow path is formed by a plurality of the flow path forming pillars or the fine grooves. [3] The device according to [1] or [2], wherein the shape of the flow path forming pillar is a cylinder, a cone, a frustum of a cone, a polygonal prism, a polygonal pyramid, or a frustum of a polygonal pyramid, and the shape of the gel-like pillar is a cylinder or a polygonal prism. [4] The device according to any one of [1] to [3], wherein the substance to be detected is an antigen and the capturing substance is an antibody against the antigen. [5] The device according to any one of [1] to [3], wherein the substance to be detected is a first antibody and the capturing substance is a second antibody specific to the first antibody. [6] The device according to any one of [1] to [5], wherein the capture substance is covalently bonded to the gel-like substance. [7] The device according to any one of [1] to [6], wherein a plurality of the gel-like pillars are provided in the detection zone in a direction orthogonal to the transfer direction of the liquid sample. [8] The device according to any one of [1] to [7], wherein a plurality of the gel-like pillars are provided in the detection zone in the transfer direction of the liquid sample in the flow path. [9] The device according to any one of [1] to [8], wherein the gel-like pillars are arranged in a lattice pattern when the detection zone is viewed from above.

[10] The device according to any one of [1] to [9], wherein the gel-like pillar is in a cylindrical shape with a diameter of 10 μm or more and 1000 μm or less.

[11] The device according to any one of [1] to

[10] , wherein the gel-like pillar is in a cylindrical shape with a height of 10 μm or more and 1000 μm or less.

[12] The device according to any one of [1] to

[11] , wherein a lid portion covering the flow path is provided.

[13] The device according to

[12] , wherein the material of the lid portion includes one or more selected from the group consisting of quartz glass, soda-lime glass, borosilicate glass, poly(meth)acrylate, polyester, polyolefin, polystyrene, polycarbonate, fluororesin, polyvinyl chloride, polyamide, and polyimide.

[14] The device according to any one of [1] to

[13] , wherein a reservoir portion or a weir portion of the liquid sample is provided on the downstream side of the detection zone.

[15] On the upstream side of the detection zone, an induction zone for guiding the liquid sample to the detection zone by capillary action is provided, The device according to any one of [1] to

[14] , wherein the bottom surface a of the detection zone is at a lower level than the bottom surface b of the induction zone.

[16] A device for capturing and detecting a substance to be detected in a liquid sample, comprising a flow path for transferring the liquid sample, The flow path has a detection zone in a part of the transfer direction of the liquid sample. In the detection zone, a gel-like first pillar is provided, which is composed of a gel substance and holds a capture substance that specifically binds to the substance to be detected. In a zone upstream or downstream of the detection zone of the flow path, a plurality of second pillars different from the first pillar are provided, or the zone is composed of microgrooves.

[17] The flow path has a guiding zone upstream of the detection zone that guides the liquid sample to the detection zone by capillary action. In the guiding zone, a plurality of second pillars are provided, or the guiding zone is composed of the microgrooves. The device according to

[16] .

[18] A chip having the device according to any one of [1] to

[17] .

[19] A substrate used for a device that captures and detects a substance to be detected in a liquid sample. A flow path for transferring the liquid sample provided on one surface of the substrate. A detection zone provided in a part of the flow path. Comprising: In the detection zone, a gel-like pillar composed of a gel substance is provided. In the gel-like pillar, a capture substance that specifically binds to the substance to be detected is held. On the upstream or downstream side of the detection zone, the flow path is formed by a plurality of flow path forming pillars or microgrooves.

[20] A long substrate. A first zone provided in a part of the longitudinal direction of the substrate, in which a gel-like first pillar composed of a gel substance and holding a capture substance that specifically binds to the substance to be detected can be arranged. Located on at least one side of the first zone in the longitudinal direction of the substrate, a plurality of second pillars different from the first pillar are provided, or a second zone composed of microgrooves. Having: [Advantages of the Invention]

[0009] According to the present invention, it is possible to provide a device that has excellent detection sensitivity and can be easily used even without using an additional device for liquid feeding.

Brief Description of the Drawings

[0010]

Figure 1

Figure 2

Figure 3

Figure 4

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Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Modes for Carrying Out the Invention

[0011] Hereinafter, embodiments of the present invention will be described with reference to the drawings. The figures are schematic and do not conform to actual dimensional ratios. In this specification, "~" indicating a numerical range represents "above" and "below", and includes both end values.

[0012] (Device) FIG. 1 is a perspective view showing an example of the configuration of the device in this embodiment. The device 100 shown in FIG. 1 is a device that captures and detects a substance to be detected in a liquid sample. The device 100 includes a substrate 101, a flow path 103 provided on one surface of the substrate 101 for transferring the liquid sample, and a detection zone 110 (first zone) provided in a part of the flow path 103. In the detection zone 110, a gel pillar (first pillar) 105 made of a gel-like substance is provided, and a capture substance 107 that specifically binds to the substance to be detected is held in the gel pillar 105. On the upstream side or the downstream side of the detection zone 110, the flow path 103 is formed by a plurality of flow path forming pillars (second pillars different from the first pillar) or fine grooves. FIG. 1 shows an example in which the flow path 103 is formed by a plurality of flow path forming pillars. In other words, a plurality of flow path forming pillars are provided in the zone on the upstream side or the downstream side of the detection zone 110 of the flow path 103, or such a zone is composed of fine grooves.

[0013] In the device 100, an induction zone 120 for guiding the liquid sample to the detection zone 110 by capillary action is provided on the upstream side of the detection zone 110. In the induction zone 120, the flow path 103 is formed by a plurality of flow path forming pillars 109. In other words, a plurality of flow path forming pillars 109 are provided in the induction zone 120 of the flow path 103. Also, in the device 100, a discharge zone 130 for discharging the liquid sample from the detection zone 110 by capillary action is provided on the downstream side of the detection zone 110. In the discharge zone 130, the flow path 103 is formed by a plurality of flow path forming pillars 111. In other words, a plurality of flow path forming pillars 111 are provided in the discharge zone 130 of the flow path 103.

[0014] (Base material) The base material 101 is specifically a substrate used as the base material of the device 100. Examples of the shape of the base material 101 include, for example, a sheet shape and a plate shape. In FIG. 1, an example of the device 100 having a sheet-shaped base material 101 is shown.

[0015] Specific examples of the planar shape of the base material 101 include polygons such as quadrilaterals, circles, and ellipses. When the base material 101 is a quadrilateral, the vertical width (length in the short side direction) of the base material 101 may be, for example, about 1 to 100 mm, and the horizontal width (length in the long side direction) of the base material 101 may be, for example, about 2 to 100 mm.

[0016] From the viewpoint of improving the strength of the device 100, the thickness of the base material 101 is, for example, 0.05 mm or more, preferably 0.1 mm or more. Also, from the viewpoint of thinning the device 100, the thickness of the base material 101 is, for example, 5 mm or less, preferably 3 mm or less.

[0017] Specific examples of the material of the base material 101 include glass such as quartz glass, soda-lime glass, and borosilicate glass; and one or more selected from the group consisting of resin materials such as poly(meth)acrylates such as polymethyl(meth)acrylate, polyesters, polyolefins, polystyrenes, polycarbonates, fluororesins, polyvinyl chlorides, polyamides, and polyimides. In terms of being able to integrally mold the base material 101 and the flow path forming pillar 109, it is preferable that the material of the base material 101 is a thermoplastic resin. Specifically, the thermoplastic resin includes one or more selected from the group consisting of polyester, polyolefin, polystyrene, polycarbonate, fluororesin, and (meth)acrylic resin. More specifically, one or more selected from the group consisting of polyethylene terephthalate (PET), cycloolefin polymer (COP), polypropylene (PP), polystyrene (PS), polycarbonate (PC), polyvinylidene fluoride (PVDF), polymethyl methacrylate (PMMA), and polyethylene (PE) are included.

[0018] In the device 100 shown in FIG. 1, an example is shown where the base material 101 is a flat plate and the gel-like pillar 105, the flow path forming pillar 109, and the flow path forming pillar 111 are arranged on its surface. However, the shape of the base material 101 is not limited to this. FIG. 13 is a perspective view showing a configuration example of a device using a base material having a recess. The basic configuration of the device 180 shown in FIG. 13 is the same as that of the device 100 shown in FIG. 1. However, in the device 180, a groove-like recess 129 with both longitudinal ends open is formed on the surface of the base material 127. In the example shown in FIG. 13, the recess 129 constitutes a flow path for moving the liquid sample. The detection zone 110 may be provided in a part of the longitudinal direction (the transfer direction of the liquid sample) of the recess 129. In the example shown in FIG. 13, the induction zone 120, the detection zone 110, and the discharge zone 130 are provided in this order from one end to the other end of the recess 129. At this time, the bottom surfaces of the gel-like pillar 105, the flow path forming pillar 109, and the flow path forming pillar 111 are located on the bottom surface of the recess 129 as shown in FIG. 13, for example. Also, the upper ends of the gel-like pillar 105, the flow path forming pillar 109, and the flow path forming pillar 111 may be located at the same level as the upper ends of the side walls defining the recess 129, that is, the upper surface of the base material 127, or may be at a different level, for example, below the upper surface of the base material 127. Also, in the guiding zone 120, the detection zone 110, and the discharge zone 130, the width of the recess 129 may be the same as shown in FIG. 13, or may be different. For example, the width of the recess 129 in the detection zone 110 may be larger than the widths of the recess 129 in the guiding zone 120 and the discharge zone 130.

[0019] (Detection zone) In the detection zone (first zone) 110, a gel pillar 105 is provided. On the other hand, as shown in FIG. 1, it is preferable that no flow path forming pillar is provided in the detection zone 110. The gel pillar 105 is composed of a gel material. Specifically, the gel material includes a polymer material having a three-dimensional network structure and becomes gel when a liquid is applied. Therefore, the gel pillar 105 may be in a state of swelling with a liquid, or may be in a state of not swelling with a liquid, that is, in a state of becoming gel when a liquid is applied to the detection zone 110 at a desired timing.

[0020] Specific examples of the material of the gel substance in the gel pillar 105 include polymer materials that form hydrogels. Examples of the polymer material include (meth)acrylates such as polyethylene glycol di(meth)acrylate, polypropylene glycol di(meth)acrylate, polyurethane (meth)acrylate, gelatin (meth)acrylate, and collagen (meth)acrylate; and one or more selected from the group consisting of poly(meth)acrylamide and collagen. Here, (meth)acrylate is at least one of acrylate and methacrylate.

[0021] Examples of the shape of the gel pillar 105 include columnar bodies such as cylinders and polygonal columns; conical bodies such as cones and polygonal pyramids; and frustum-shaped bodies such as frustums of cones and frustums of polygonal pyramids. These do not necessarily have geometrically accurate shapes, and may have shapes with rounded corners or shapes with fine irregularities on the surface. From the viewpoint of improving formability, the shape of the gel pillar 105 is preferably a cylinder or a polygonal prism.

[0022] When the gel pillar 105 is cylindrical, the diameter of the gel pillar 105 is preferably 10 μm or more, more preferably 50 μm or more, from the viewpoint of improving formability. Also, from the viewpoint of improving formability, the diameter of the gel pillar 105 is preferably 1000 μm or less, more preferably 500 μm or less. For the diameter of the gel pillar 105, for example, five arbitrary gel pillars 105 can be selected from the detection zone 110, and the average value of the diameters of the bottom or top surfaces of the five selected gel pillars 105 can be adopted.

[0023] When the gel pillar 105 is cylindrical, the height of the gel pillar 105 is preferably 10 μm or more, more preferably 30 μm or more, from the viewpoint of improving formability. Also, from the viewpoint of improving formability, the height of the gel pillar 105 is preferably 1000 μm or less, more preferably 500 μm or less. For the height of the gel pillar 105, for example, five arbitrary gel pillars 105 can be selected from the detection zone 110, and the average value of the heights of the five selected gel pillars 105 can be adopted.

[0024] In the detection zone 110, the number of gel pillars 105 may be one or two or more. In the detection zone 110, one gel pillar 105 may be provided in the direction orthogonal to the transfer direction X of the liquid sample (w in FIG. 1: the width direction of the flow path 103), or two or more gel pillars 105 may be provided. From the viewpoint of improving the detection sensitivity and measurement accuracy of the analyte, as shown in FIG. 1, in the detection zone 110, preferably a plurality of gel pillars 105 are provided in the direction orthogonal to the transfer direction X of the liquid sample. Also, in the detection zone 110, it is preferable that a plurality of gel pillars 105 are provided in the transfer direction X (the extending direction of the flow path 103) of the liquid sample.

[0025] The planar arrangement of the plurality of gel pillars 105 may be regular or irregular. From the viewpoint of improving the detection sensitivity and measurement accuracy of the substance to be detected, when the detection zone 110 is viewed from above, the gel pillars 105 are preferably arranged in a lattice pattern. More specifically, as the lattice pattern, a square lattice; an oblique lattice such as a hexagonal lattice can be mentioned.

[0026] When a plurality of gel pillars 105 are provided, the distance between adjacent gel pillars 105 in the detection zone 110, that is, the closest distance between the gel pillars 105, is 0 μm or more and is appropriately set according to the shape of the gel pillars 105. For example, when the gel pillar 105 is a columnar body, the closest distance is preferably more than 0 μm, and when the gel pillar 105 is a conical body or a frustum of a cone, the closest distance is 0 μm or more. From the viewpoint of more efficiently moving a liquid such as a liquid sample in the detection zone 110, the closest distance is, for example, 5 μm or more, preferably 10 μm or more. Further, from the viewpoints that the contact area between the liquid sample and the gel pillar 105 increases, and thus the capillary force increases, making it easier to move the liquid sample, and that the production is easier, the closest distance is preferably 1000 μm or less, more preferably 500 μm or less.

[0027] Here, the "distance between adjacent gel pillars 105" is the distance defined by their peripheral surfaces on the line segment connecting the center points in the top view of the two gel pillars 105 located closest to each other. Specifically, as the distance between adjacent gel pillars 105, five distances between any adjacent gel pillars 105 can be selected from the detection zone 110, and the average value of the five selected distances can be adopted. FIG. 3(a), FIG. 3(b) and FIG. 4 are top views showing arrangement examples of the gel pillars 105. The planar shape of the gel pillars 105 is circular in FIG. 3(a) and FIG. 4, and rectangular in FIG. 3(b). For example, in an array where cylinders are arranged in a square lattice, the above-mentioned nearest neighbor distance is the distance of the arrow in Fig. 3(a). In an array where cylinders are arranged in a hexagonal lattice, the above-mentioned nearest neighbor distance is the distance of the arrow in Fig. 4. For example, as shown in Fig. 3(a), the smaller the distance between adjacent gel-like pillars 105, the smaller the gap between adjacent gel-like pillars 105, the larger the number of gel-like pillars 105 per unit area in top view, the larger the contact area between the gel-like pillars 105 and the liquid sample, and the larger the amount of the detected substance bound by the capture substance 107. As a result, the detection sensitivity of the detected substance can be increased. Regarding the above-mentioned detection sensitivity, more specifically, when the area of the detection zone 110 used for detecting the detected substance captured by the gel-like pillar 105 is larger than the size of the gel-like pillar 105 in top view, by reducing the above-mentioned nearest neighbor distance, the number of gel-like pillars 105 per unit area increases, and the intensity of the detection signal per unit area can be increased. Also, in top view, the larger the number of gel-like pillars 105 per unit area, the higher the probability that the gel-like pillar 105 and the detected substance come into contact, and the detected substance can be captured by the gel-like pillar 105 more efficiently. As a result, the intensity of the detection signal per unit area can be increased. On the other hand, the larger the distance between adjacent gel-like pillars 105, the easier the molding.

[0028] The capture substance 107 is held on the gel-like pillar 105. The capture substance 107 may be chemically immobilized on the gel-like pillar 105 or physically immobilized. Also, the capture substance 107 may be encapsulated in the gel-like pillar 105 or supported on the surface of the gel-like pillar 105. From the viewpoint of more stably retaining the capture substance 107 in the gel pillar 105, the capture substance 107 is preferably covalently bonded to the gel substance in the gel pillar 105. At this time, the capture substance 107 may be directly bonded to the gel pillar 105, or may be bonded to an intervening molecule such as a spacer molecule bonded to the gel pillar 105. The intervening molecule preferably contains glycols such as polyethylene glycol, ethers, amines, esters, amides, alcohols, carboxylic acids, etc.

[0029] The capture substance 107 is selected from substances that specifically bind to the substance to be detected. For example, when the substance to be detected is an antigen, the capture substance 107 may be an antibody against the antigen or an antigen-binding fragment thereof, preferably an antibody. Here, the antigen may be a substance having immunogenicity alone or a hapten. Also, the substance to be detected may be a first antibody, and the capture substance may be a second antibody specific to the first antibody. When the capture substance 107 is an antibody or an antigen-binding fragment thereof, the antibody may be a polyclonal antibody or a monoclonal antibody. When the capture substance 107 is an antibody or an antigen-binding fragment thereof, the substance to be detected may be a substance capable of antigen-antibody reaction with the antibody, and examples thereof include various pathogens and various clinical markers. More specifically, the substances to be detected include virus antigens such as influenza virus, norovirus, adenovirus, respiratory syncytial virus (RSV), hepatitis A virus (HAV), hepatitis B surface antigen (HBs), and human immunodeficiency virus (HIV); bacterial antigens such as methicillin-resistant Staphylococcus aureus (MRSA), group A streptococcus, group B streptococcus, and Legionella bacteria; toxins produced by bacteria, etc.; hormones such as mycoplasma, Chlamydia trachomatis, and human chorionic gonadotropin; C-reactive protein, myoglobin, cardiac troponin, various tumor markers, pesticides, and environmental hormones. When the substances to be detected are items such as influenza virus, norovirus, C-reactive protein, myoglobin, and cardiac troponin that require urgent detection and treatment measures, their usefulness is even greater. The substance to be detected is usually in a suspended or dissolved state in a liquid sample. The liquid sample may be, for example, a sample in which the substance to be detected is suspended or dissolved in a buffer solution.

[0030] The substance to be detected is not limited to an antigen, and may be selected from the group consisting of, for example, proteins and peptides such as enzymes and antibodies; nucleic acids; polysaccharides; and glycoproteins. And the capture substance 107 may be any substance having specificity for these substances to be detected. For example, the capture substance 107 may be selected from the group consisting of proteins, nucleic acids, polysaccharides, and glycoproteins.

[0031] The device 100 is shown with the bottom surface a of the detection zone 110 at the same level as the bottom surface b of the induction zone 120. However, from the viewpoint of further enhancing the detection sensitivity of the substance to be detected, the bottom surface a of the detection zone 110 may be at a lower level than the bottom surface b of the induction zone 120. Also, the bottom surface a of the detection zone 110 may be at a lower level than the bottom surface c of the discharge zone 130.

[0032] (Induction zone) In the induction zone (second zone) 120, from the viewpoint of surely guiding the liquid sample to the detection zone 110 by capillary action, preferably a plurality of flow path forming pillars 109 are provided. The planar arrangement of the plurality of flow path forming pillars 109 may be regular or irregular. From the same viewpoint, when the induction zone 120 is viewed from above, the flow path forming pillars 109 are preferably arranged in a lattice pattern. As the lattice pattern, more specifically, a square lattice; an oblique lattice such as a hexagonal lattice can be mentioned.

[0033] Examples of the shape of the flow path forming pillar 109 include columnar bodies such as cylinders and polygonal columns; conical bodies such as cones and polygonal pyramids; and frustum shapes such as frustum of cones and frustum of polygonal pyramids. These do not necessarily have to be geometrically exact shapes, and may be shapes with rounded corners or shapes with fine irregularities on the surface. From the viewpoint of improving moldability, the shape of the flow path forming pillar 109 is preferably a cylinder, a cone, a frustum of a cone, a polygonal column, a polygonal pyramid or a frustum of a polygonal pyramid, more preferably a cylinder, a cone or a frustum of a cone, and even more preferably a frustum of a cone. The shapes of the plurality of flow path forming pillars 109 may be the same or different. From the viewpoint of more reproducibly producing a desired fine concavo-convex structure, the shapes of the plurality of flow path forming pillars 109 are preferably the same.

[0034] When the flow path forming pillar 109 is a frustum of a cone, the diameter of the bottom surface of the flow path forming pillar 109 is preferably 5 μm or more, more preferably 10 μm or more, from the viewpoint of improving moldability. Also, from the viewpoint of improving moldability, the diameter of the bottom surface of the flow path forming pillar 109 is preferably 1000 μm or less, more preferably 500 μm or less. The diameter of the bottom surface of the flow path forming pillar 109 can be, for example, by selecting 5 arbitrary flow path forming pillars 109 from the induction zone 120 and adopting the average value of the diameters of the bottom surfaces of the selected 5 flow path forming pillars 109.

[0035] When the flow path forming pillar 109 is a frustum of a cone, the height of the flow path forming pillar 109 is preferably 10 μm or more, more preferably 25 μm or more, from the viewpoint of improving moldability. Also, from the viewpoint of improving moldability, the height of the flow path forming pillar 109 is preferably 1000 μm or less, more preferably 300 μm or less. For the height of the flow path forming pillar 109, for example, five arbitrary flow path forming pillars 109 can be selected from the induction zone 120, and the average value of the heights of the selected five flow path forming pillars 109 can be adopted.

[0036] The distance between adjacent flow path forming pillars 109 in the induction zone 120, that is, the closest distance between the flow path forming pillars 109, is appropriately set according to the shape of the flow path forming pillar 109, and is, for example, about 0 to 500 μm. For example, when the flow path forming pillar 109 is a columnar body, the closest distance is preferably more than 0 μm, and when the flow path forming pillar 109 is a conical body or a frustum of a cone, the closest distance is 0 μm or more. Also, the closest distance may be, for example, 0.1 μm or more, or may be, for example, 2 μm or more. The upper limit of the closest distance is preferably 500 μm or less, more preferably 100 μm or less, and even more preferably 50 μm or less. Thereby, the contact area between the liquid sample and the base material 101 and the flow path forming pillar 109 increases, and the capillary force increases, so that it becomes easy to move the liquid sample.

[0037] Here, the "distance between adjacent flow path forming pillars 109" is the distance defined by their peripheral surfaces on the line segment connecting the center points in the top view of the two closest flow path forming pillars 109. As the distance between adjacent flow path forming pillars 109, specifically, five distances between arbitrary adjacent flow path forming pillars 109 can be selected from the induction zone 120, and the average value of the selected five distances can be adopted. For example, when the closest distance between the conical or frustum-shaped flow path forming pillars arranged in a hexagonal lattice is 0 μm, adjacent flow path forming pillars 109 can be arranged without gaps, as shown in FIG. 5 for example. FIG. 5 is a top view showing an arrangement example of the flow path forming pillars 109. By arranging the flow path forming pillars 109 without gaps, the number of flow path forming pillars 109 per unit area increases, and the capillary force further increases, making it easier to transfer the liquid sample.

[0038] Specific examples of the material of the flow path forming pillar 109 include those described above as the material of the base material 101. The material of the flow path forming pillar 109 may be the same as or different from the material of the base material 101. Also, for example, the flow path forming pillar 109 and the base material 101 may be composed of a single member. That is, the device 100 may not have an interface between the base material 101 and the flow path forming pillar 109.

[0039] (Discharge zone) In the discharge zone (second zone) 130, the flow path 103 is formed by a plurality of flow path forming pillars 111. In other words, a plurality of flow path forming pillars 111 are provided in the discharge zone 130. The configurations of the discharge zone 130 and the flow path forming pillars 111 can be, for example, in accordance with those of the induction zone 120 and the flow path forming pillars 109 respectively.

[0040] Although not shown in FIG. 1, from the viewpoint of promoting discharge in the discharge zone 130, the discharge zone 130 of the device 100 may be discharged by overlapping an absorption pad. Examples of the material of the absorption pad include paper and porous bodies. Regarding the arrangement of the absorption pad, for example, it may be arranged by overlapping the base material 101 in at least a part of the discharge zone 130, or may be arranged adjacent to the base material 101.

[0041] The device 100 shown in FIG. 1 is not provided with a lid, but the device may be provided with a lid that covers the flow path. Figure 2 is a cross-sectional view showing an example of another configuration of the device in the present embodiment. The basic configuration of the device 150 shown in FIG. 2 is the same as that of the device 100 described above with reference to FIG. 1, except that the lid portion 113 facing the base material 101 is provided so as to cover the flow path 103. By providing the lid portion 113, it is possible to suppress the drying of the gel-like pillars 105 in the detection zone 110. Also. In the induction zone 120 provided with the flow path forming pillar 109 and the discharge zone 130 provided with the flow path forming pillar 111, the flow of the liquid sample due to capillary action can be made more stable. The lid portion 113 may cover the entire base material 101 or may cover a part of the base material 101.

[0042] From the viewpoint of improving the strength of the device 150, the thickness of the lid portion 113 is, for example, 5 μm or more, preferably 10 μm or more. Also, from the viewpoint of thinning the device 150, the thickness of the lid portion 113 is, for example, 1000 μm or less, preferably 200 μm or less.

[0043] From the viewpoint of improving the visibility from the upper part of the device 150, the lid portion 113 is preferably made of a transparent material. More specifically, the material of the lid portion 113 includes glass such as quartz glass, soda lime glass, borosilicate glass; and one or more selected from the group consisting of resins such as poly(meth)acrylate, polyester, polyolefin, polystyrene, polycarbonate, fluororesin, polyvinyl chloride, polyamide, and polyimide.

[0044] In the above example, the configuration in which the induction zone 120, the detection zone 110, and the discharge zone 130 are provided in this order from the upstream side of the device, that is, the liquid sample introduction side, to the downstream side has been described as an example. However, for the device, at least one of the induction zone 120 and the discharge zone 130 may be provided. Also, the device may be provided with a region other than the above-described regions. For example, a sample introduction unit may be provided upstream of the detection zone 110, more specifically, upstream of the guiding zone 120, in the device. Also, FIGS. 6(a) to 6(d) are cross-sectional views showing arrangement examples of the device. As shown in FIG. 6(a), the device may be provided with a flat portion 115 where no flow path forming pillar is provided, between the detection zone 110 and the discharge zone 130. Also, as shown in FIG. 6(c), FIG. 6(d) or FIG. 6(b), the device may be provided with a liquid sample storage portion 119 or storage portion 121 or a weir portion 117 downstream of the detection zone 110, more specifically, upstream of the discharge zone 130. None of the storage portion 119 or storage portion 121 and the weir portion 117 are provided with flow path forming pillars.

[0045] In FIG. 6(b), in the weir portion 117, its height (the thickness of the base material 101) gradually increases from the detection zone 110 toward the discharge zone 130, and a step is formed at the boundary with the discharge zone 130. Due to the presence of this step, an effect of preventing the backflow of the liquid sample from the discharge zone 130 to the detection zone 110, that is, a weir effect, is exerted. Also, in the storage portion 119 of FIG. 6(c) and the storage portion 121 of FIG. 6(d), a region is provided where the bottom surface of the base material 101 is located at a level lower than the bottom surface of the base material 101 in the detection zone 110. Thereby, the moving speed of the liquid in the detection zone 110 can be adjusted. Also, the configurations of FIGS. 6(a) to 6(d) can be appropriately combined to form a desired device, and a more preferable flow path configuration can be obtained according to the type and amount of the liquid introduced into the flow path 103, and the moving speed of the liquid can be made more suitable.

[0046] In the above example, in the induction zone 120 and the discharge zone 130, an example is shown where the flow path 103 is constituted by a plurality of flow path forming pillars 109 and a plurality of flow path forming pillars 111 respectively (an example where the plurality of flow path forming pillars 109 and the plurality of flow path forming pillars 111 are provided in the flow path 103), but the flow path 103 may be constituted by microgrooves. FIG. 7 is a top view showing a configuration example of a device having microgrooves. In the device 140 shown in FIG. 7, it is an example where the flow paths in the induction zone 120 and the discharge zone 130 shown in FIG. 1 are constituted by the microgroove 123 and the microgroove 125 respectively. The widths of the microgroove 123 and the microgroove 125 in the direction orthogonal to the transfer direction of the liquid sample are preferably 2 μm or more, also preferably 2000 μm or less, and more preferably 1000 μm or less from the viewpoint that the contact area between the liquid sample and the flow path 103 increases, thereby increasing the capillary force and making it easier to move the liquid sample. The widths of the microgroove 123 and the microgroove 125 may be the same or different. Moreover, as a device having microgrooves, for example, a shape is exemplified in which the volumes of the microgroove 123 and the microgroove 125 are 1000 μm in width × 20 mm in length × 1000 μm in depth, and the volume of the detection zone 110 is 2000 μm in width × 10 mm in length × 1000 μm in depth, and the gel-like pillar 105 is arranged in the detection zone 110. In the example of FIG. 7, the detection zone 110 is a region wider than the microgroove 123 and the microgroove 125 in the direction orthogonal to the transfer direction of the liquid sample, and a plurality (four in the figure) of gel-like pillars 105 are regularly arranged in the detection zone 110.

[0047] In addition, in the above examples, the description focused on the example where a plurality of gel pillars 105 were provided in the detection zone 110. However, only one gel pillar 105 may be provided in the detection zone 110. Also, from the viewpoints of easier production, simpler detection system, and improved fluidity of the liquid, it is preferable to provide one gel pillar 105 in the detection zone 110. FIGS. 8 and 9 are perspective views showing configuration examples of devices in which one gel pillar 105 is provided in the detection zone 110. The basic configurations of the device 160 shown in FIG. 8 and the device 170 shown in FIG. 9 are both the same as the configuration of the device 100 described above with reference to FIG. 1, but the difference is that one columnar and one rectangular parallelepiped gel pillar 105 are provided in the detection zone 110, respectively. When one gel pillar 105 is provided in the detection zone 110, in a top view, for example, the area of the gel pillar 105 may be larger than the areas of the flow path forming pillars 109 and 111.

[0048] Next, taking a device having the flow path forming pillars 109 and 111 as an example, a method for manufacturing the device will be described. The device can be obtained, for example, by forming the gel pillar 105 and at least one of the flow path forming pillars 109 and 111 on one surface of the base material 101 in a predetermined order.

[0049] As methods for forming the flow path forming pillars 109 and 111, for example, imprinting such as thermal imprinting and UV imprinting; injection molding; pattern formation on a UV curable resin by photolithography, soft lithography using a UV curable resin pattern formed by photolithography as a mold, pattern formation by etching using a UV curable resin pattern formed by photolithography, etc.; mechanical cutting; and Laser processing and the like can be mentioned. Among these, hot embossing and injection molding for thermoplastic resins are suitable as methods for performing precise processing at low cost. Specific examples of the thermoplastic resin include those described above as the material of the base material 101. When the shapes of the flow path forming pillars 109 and 111 are cones, in the case of a processing method using a mold such as embossing or injection molding, since the upper part of the cone is thinner than the bottom surface, the volume to be cut out during mold production is less than that for producing a pillar with the same bottom surface, and the mold can be produced at low cost. In this case, it becomes possible to detect the analyte in the liquid sample at a lower cost.

[0050] As a method for forming the gel-like pillar 105, after applying a liquid containing a UV curable resin and a polymerization initiator between a pair of transparent materials (for example, glass) opposed via spacers at a predetermined interval, a photomask is provided in the formation region of the gel-like pillar 105, and by irradiating with UV light, the resin can be selectively cured in the region where the photomask is provided to form the pattern of the gel-like pillar 105. By arranging the obtained gel-like pillar 105 at a predetermined position on the base material 101 provided with the flow path forming pillars 109 and 111, a device can be obtained. Also, at this time, a liquid containing a gelling agent may be applied to a predetermined region on the surface of the base material 101 provided with at least one of the flow path forming pillars 109 and 111, and then the gel-like pillar 105 may be formed according to the above-described procedure, so that the gel-like pillar 105 may be directly formed on the base material 101.

[0051] (Detection method) In this embodiment, the device can be suitably used for immunoassay. Since the device in this embodiment is provided with at least one of a detection zone 110 having a gel pillar 105 and an induction zone 120 having a flow path forming pillar 109 or a discharge zone 130 having a flow path forming pillar 111, a substance to be detected in a liquid sample can be stably detected with excellent sensitivity. Further, since the device in this embodiment can be used even when no additional device such as a liquid feeding pump is used, the substance to be detected can be easily detected.

[0052] In this embodiment, the detection method using the device can be performed, for example, by the sandwich method. At this time, specifically, the detection method is A step of introducing a liquid sample containing a substance to be detected upstream of the detection zone 110, specifically, on or upstream of the induction zone 120, guiding it to the detection zone 110 by capillary action, and capturing the substance to be detected by the gel pillar 105 by specific interaction between the substance to be detected and the capture substance 107 (step 11); After the capturing step, a step of introducing a liquid containing a labeled antibody that specifically binds to the substance to be detected into the detection zone 110 and capturing the labeled antibody by the gel pillar 105 where the substance to be detected is captured (step 12); and A step of detecting or quantifying the substance to be detected in the liquid sample by detecting the labeled antibody captured by the gel pillar 105 (step 13). is included. Further, between at least one of step 11 and step 12 and between step 12 and step 13, a step of introducing a buffer solution or the like upstream of the detection zone 110, specifically, upstream of the induction zone 120, and flushing the detection zone 110 (step 14) may be further performed.

[0053] For the labeled antibody in step 12, labeled antibodies used in immunoassays such as enzyme-linked immunosorbent assay (ELISA) and fluorescence immunoassay can be used. Examples of the labeled antibody include a fluorescently labeled antibody and an enzyme-labeled antibody. In addition, in step 13, a detection method corresponding to the type of labeled antibody used in step 12 can be used. For example, when a fluorescently labeled antibody is used in step 12, in step 13, the substance to be detected can be detected or quantified by measuring the presence or absence or intensity of fluorescence in the gel pillar 105. When an enzyme-labeled antibody is used in step 12, in step 13, a substrate for the enzyme immobilized on the labeled antibody is introduced into the detection zone 110, and the substance to be detected can be detected or quantified by measuring the presence or absence of color development based on the substrate, absorbance, presence or absence of fluorescence, fluorescence intensity, presence or absence of chemiluminescence, chemiluminescence intensity, etc.

[0054] (Chip) In this embodiment, the chip has the device in the above-described embodiment. The chip may be composed of the device, or may further have other members. Specific examples of the other members include a housing that houses or holds the device.

[0055] (Substrate) In this embodiment, the substrate is used for the device in the above-described embodiment. For the substrate, the configuration described above for the base material 101 can be appropriately used. The substrate is long and has a first zone provided in a part of its longitudinal direction and a second zone located on at least one side (both sides in the case of the base material 101 shown in FIG. 1) of the first zone. By disposing the gel-like pillar 105 in the first zone, the detection zone 110 of the device 100 can be made to function. On the other hand, in the second zone, a plurality of flow path forming pillars (pillars different from the gel-like pillar 105), 109 and 111, are provided, or since the second zone is composed of fine grooves, the guiding zone 120 and the discharging zone 130 of the device 100 can be made to function.

[0056] As described above, the embodiments of the present invention have been described with reference to the drawings, but these are examples of the present invention, and various configurations other than the above can also be adopted.

Example

[0057] Hereinafter, the present embodiment will be specifically described with reference to examples and comparative examples, but the present embodiment is not limited to these examples.

[0058] (Examples 1 and 2) In this example, a device in which an anti-CRP antibody was immobilized on a gel-like pillar was prepared, and CRP (C-reactive protein) in a liquid sample was detected.

[0059] (Preparation of Device) A device having the schematic structure shown in FIG. 2 was prepared. 1. Preparation of the guiding zone 120 and the discharging zone 130 A substrate 101 having flow path forming pillars 109 and flow path forming pillars 111 was prepared by the following procedure. The shapes and arrangements of the flow path forming pillars in the guiding zone 120 and the discharging zone 130 were both the same. A polymethyl methacrylate sheet (manufactured by Sumitomo Chemical Acrylic Sales Co., Ltd., film thickness: approximately 200 μm) was subjected to hot embossing under the conditions of pressing a mold against it, a heating temperature of 160 °C, a pressure of 5.5 MPa, and a pressurization time of 3 minutes, to produce a base material 101 in which frustum-shaped protrusions with a bottom diameter of the pillar (hereinafter, also referred to as "diameter") of 60 μm and a height of the pillar (hereinafter, also referred to as "height") of 90 μm are arranged in a hexagonal lattice arrangement with an average distance between the centers of the protrusions of 62 μm, as shown in FIGS. 10(a) and 10(b), and the gap between the induction zone 120 and the discharge zone 130 is 4.5 mm. Here, for the mold used in the hot embossing, frustum-shaped holes with a diameter of the hole entrance of 60 μm and a depth of 90 μm are arranged in a hexagonal lattice arrangement with an average distance between the centers of the holes of 62 μm, and a nickel mold with a gap corresponding to that between the induction zone 120 and the discharge zone 130 of the base material 101 being 4.5 mm was used. The holes were produced by machining. The planar shape of the base material 101 was set to a width of 5 mm × a length of 30 mm, the planar shape of the induction zone 120 was set to a width of 5 mm × a length of 4.5 mm, and the planar shape of the discharge zone 130 was set to a width of 5 mm × a length of 21 mm. FIGS. 10(a) and 10(b) are, respectively, a top view showing an optical microscope of the flow path forming pillars 109 and 111 in the obtained base material 101 and a perspective view showing an SEM image. In FIGS. 10(a) and 10(b), the flow path forming pillars adjacent to each other in the horizontal direction are arranged without a gap, and the flow path forming pillars adjacent to each other in the diagonal direction in FIGS. 10(a) and 10(b) are arranged almost without a gap.

[0060] 2. Preparation of Gel Pillars 105 Gel pillars 105 were formed on the base material 101 obtained in 1. by the following procedure. The planar shape of the detection zone 110 was set to a width of 5 mm × a length of 4.5 mm. 2.1 Preparation of Polymer Solution For Examples 1 and 2, the following polymer solutions were prepared respectively. (a) Example 1 A solution obtained by diluting polyethylene glycol diacrylate (PEGDA, number average molecular weight 575, manufactured by Aldrich) with phosphate buffered saline (PBS) was added with 2-hydroxy-2-methylpropiophenone as a photoinitiator and anti-CRP antibody as an antibody to prepare a polymer solution (antibody concentration of about 100 μg / mL, 20% PEGDA, 0.55% initiator). (b) Example 2 A solution obtained by diluting polyethylene glycol diacrylate (PEGDA, number average molecular weight 575, manufactured by Aldrich) with phosphate buffered saline (PBS) was added with 2-hydroxy-2-methylpropiophenone as a photoinitiator and an anti-CRP antibody modified with a PEG-Ac linker (PEG: molecular weight 2000) (PEG acrylated CRP antibody, Ac-PEG-Ab) as an antibody to prepare a polymer solution (antibody concentration of about 300 μg / mL, 20% PEGDA, 0.54% initiator). Ac: Acrylic group PEG: Polyethylene glycol group Ab: Antibody molecule

[0061] 2.2 Gelation The polymer solution of each example was supplied to the gap between the induction zone 120 and the discharge zone 130 in the base material 101 obtained in 1., and after placing a cover glass (thickness of about 0.15 mm), a photomask (circular holes with a diameter of 400 μm arranged in a square lattice with a center-to-center distance of 800 μm) was placed on the cover glass, and using an exposure device (UIV270, manufactured by USHIO Inc.), the polymer solution of Example 1 was irradiated with light for 90 seconds, and the polymer solution of Example 2 was irradiated with light for 60 seconds to prepare gel-like pillars 105. Then, the uncured polymer solution was washed away with 2% Triton x-100 / PBS.

[0062] 3. Disposal of absorption pad At a position 10 mm from the end of the formation region (discharge zone 130) of the flow path forming pillar 111 on the base material 101 on which the gel-like pillar 105 is formed, an absorption pad (PVA sponge, A-3150HP, manufactured by Ion Co., thickness approximately 0.6 mm) cut to a size of 5 mm in width × 40 mm in length was overlaid and fixed with tape.

[0063] (Immunoassay) Using the devices obtained in each example, an immunoassay was performed according to the following procedure. 1. A 10 μL aqueous solution (CRP, concentration 10 μg / mL) obtained by diluting an antigen with 2% TritonX100 / PBS was dropped onto the induction zone 120 on the base material 101, and waited for 3 minutes. 2. A 10 μL aqueous solution (FITC (fluorescein isothiocyanate)-labeled anti-CRP antibody, 20 μg / mL) obtained by diluting a fluorescently labeled antibody with 2% TritonX100 / PBS was dropped onto the induction zone 120 on the base material 101, and waited for 1 minute. This operation was repeated 2 more times (3 times in total). 3. 10 μL of 2% TritonX100 / PBS was dropped onto the induction zone 120 on the base material 101, and waited for 1 minute. This operation was repeated 1 more time (2 times in total). 4. After confirming that the aqueous solution had generally flowed off the base material 101 and was absorbed by the absorption pad, the absorption pad was removed, and the region where the gel-like pillar 105 was formed on the base material 101 was observed using a fluorescence microscope (IX71, manufactured by Olympus). The evaluation results of Example 1 and Example 2 are shown in FIGS. 11(a) and 11(b) respectively. FIGS. 11(a) and 11(b), and FIGS. 12(a) and 12(b) described later show fluorescence microscope images of the gel-like pillar 105.

[0064] (Comparative Example 1) In Example 1, in the procedure of 1. of the immunoassay, evaluation was carried out according to Example 1 except that 2% TritonX100 / PBS was used instead of the aqueous solution in which the antigen was diluted. The results are shown in FIG. 12(a).

[0065] (Comparative Example 2) In Example 2, in the procedure of 1. of the immunoassay, evaluation was carried out according to Example 2, except that 2% TritonX100 / PBS was used instead of the aqueous solution in which the antigen was diluted. The results are shown in Fig. 12(b).

[0066] From Figs. 11(a), 11(b), 12(a) and 12(b), it can be seen that in each example, CRP is captured by the gel pillar 105 through specific binding with the anti-CRP antibody, and this can be stably detected.

[0067] This application claims the priority based on Japanese Patent Application No. 2021-025325 filed on February 19, 2021, and incorporates all of its disclosures herein.

Explanation of Signs

[0068] 100 Device 101 Substrate 103 Flow path 105 Gel pillar (first pillar) 107 Capture substance 109 Flow path forming pillar (second pillar) 110 Detection zone (first zone) 111 Flow path forming pillar 113 Lid part 115 Flat part 117 Weir part 119 Storage part 120 Induction zone (second zone) 121 Storage part 123 Microgroove 125 Microgroove 127 Substrate 129 Concave part 130 Discharge zone 140 Device 150 Device 160 Device 170 Device 180 Device

Claims

1. A device for capturing and detecting a substance to be detected in a liquid sample, comprising: a substrate; a flow path provided on one surface of the substrate for transferring the liquid sample; a detection zone provided in a part of the flow path; wherein gel-like pillars composed of a gel substance are provided in the detection zone, and in the gel-like pillars, a capture substance that specifically binds to the substance to be detected is covalently bound and immobilized to a spacer molecule covalently bound to the gel substance and held therein, when the detection zone of one of the flow paths is viewed from above, a plurality of the gel-like pillars are arranged in a lattice pattern, and on the upstream side or downstream side of the detection zone, the flow path is formed by a plurality of flow path forming pillars or fine grooves (excluding a device in which the capture substance is an antibody and polystyrene beads coated with the antibody are contained in the gel-like pillars).

2. An induction zone for guiding the liquid sample to the detection zone by capillary action is provided on the upstream side of the detection zone, and in the induction zone, the flow path is formed by a plurality of the flow path forming pillars or the fine grooves. The device according to claim 1.

3. The shape of the flow path forming pillar is a cylinder, a cone, a frustum of a cone, a prism, a pyramid or a frustum of a pyramid, and the shape of the gel-like pillar is a cylinder or a prism. The device according to claim 1 or 2.

4. The device according to any one of claims 1 to 3, wherein the substance to be detected is an antigen and the capture substance is an antibody against the antigen.

5. The device according to any one of claims 1 to 3, wherein the substance to be detected is a first antibody and the capture substance is a second antibody specific to the first antibody.

6. The device according to any one of claims 1 to 5, wherein the gel-like pillar is cylindrical with a diameter of 10 μm or more and 1000 μm or less.

7. The device according to any one of claims 1 to 6, wherein the gel-like pillar is cylindrical with a height of 10 μm or more and 1000 μm or less.

8. The device according to any one of claims 1 to 7, wherein a lid portion covering the flow path is provided.

9. The device according to claim 8, wherein the material of the lid portion comprises one or more selected from the group consisting of quartz glass, soda-lime glass, borosilicate glass, poly(meth)acrylate, polyester, polyolefin, polystyrene, polycarbonate, fluororesin, polyvinyl chloride, polyamide, and polyimide.

10. The device according to any one of claims 1 to 9, wherein a reservoir or a weir for the liquid sample is provided on the downstream side of the detection zone.

11. An induction zone for guiding the liquid sample to the detection zone by capillary action is provided on the upstream side of the detection zone, The device according to any one of claims 1 to 10, wherein the bottom surface a of the detection zone is at a lower level than the bottom surface b of the induction zone.

12. A device for capturing and detecting a substance to be detected in a liquid sample, Comprising a flow path for transferring the liquid sample, The flow path has a detection zone in a part of the transfer direction of the liquid sample, In the detection zone, a gel-like first pillar is provided, which is composed of a gel substance and in which a capture substance that specifically binds to the substance to be detected is covalently bonded to a spacer molecule covalently bonded and immobilized in the gel substance, and a plurality of second pillars different from the first pillar are provided in a zone upstream or downstream of the detection zone of the flow path, or the zone upstream or downstream of the detection zone of the flow path is composed of microgrooves. A device in which when the detection zone of one of the flow paths is viewed from above, a plurality of the first pillars are arranged in a lattice pattern (excluding a device in which the capture substance is an antibody and polystyrene beads coated with the antibody are contained in the gel-like pillar).

13. The flow path has an induction zone for guiding the liquid sample to the detection zone by capillary action on the upstream side of the detection zone, The device according to claim 12, wherein the plurality of second pillars are provided in the induction zone, or the induction zone is composed of the microgrooves.

14. A chip having the device according to any one of claims 1 to 13.

15. A substrate used for a device for capturing and detecting a substance to be detected in a liquid sample, A flow path for transferring the liquid sample provided on one surface of the substrate, A detection zone provided in a part of the flow path, Comprising The detection zone is provided with gel pillars made of a gel-like substance, and in the gel pillars, a capture substance that specifically binds to the substance to be detected is covalently bonded and immobilized to a spacer molecule that is covalently bonded to the gel-like substance and is held therein, when the detection zone of one of the flow paths is viewed from above, a plurality of the gel pillars are arranged in a lattice pattern, on the upstream side or the downstream side of the detection zone, the flow path is formed by a plurality of flow path forming pillars or fine grooves (excluding a device in which the capture substance is an antibody and polystyrene beads coated with the antibody are included in the gel pillars).

16. A long substrate used for a device that captures and detects a substance to be detected in a liquid sample, a first zone provided in a part of the longitudinal direction of the substrate, configured of a gel-like substance, and in which a gel-like first pillar in which a capture substance that specifically binds to the substance to be detected is covalently bonded and immobilized to a spacer molecule that is covalently bonded to the gel-like substance can be arranged, and a second zone that is located on at least one side of the first zone in the longitudinal direction of the substrate and is provided with a plurality of second pillars different from the first pillar or is configured of fine grooves, having a substrate (excluding a device in which the capture substance is an antibody and polystyrene beads coated with the antibody are included in the gel pillars), wherein when one of the first zones is viewed from above, a plurality of the first pillars are arranged in a lattice pattern.

Images