JP2007212215A - Porous carrier and method of manufacturing same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To cope with the problem that a process for assembling individual members and a process for inspecting whether they remain precisely assembled when an electrode and a porous film are necessary in the measurement of a material to be inspected in a liquid sample. <P>SOLUTION: This porous carrier is prepared by forming an electroconductive layer on the upper surface of an insulating substrate and using the electroconductive layer forming insulating substrate as a support to fix and form the porous film to the upper surface of the electroconductive layer. By this method, the porous carrier capable of performing various electrochemical analysises by using an electrode pattern as the electroconductive layer or forming the porous film to a part of the electroconductive layer is provided corresponding to the use of the porous carrier. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a porous carrier comprising fine pores having an electrically conductive layer formed on an insulating substrate, and a method for producing the carrier.

  Polymer membrane filters composed of porous membranes with fine pores are widely used as liquid separation filtration, separation of biological materials, adsorption fixation, and chromatographic substrates, contributing to the medical, food industry, general industry, semiconductor industry, etc. is doing. There are many uses for membrane filters, but the main usage is as follows.

  When used in filtration applications, fix the membrane filter with a jig, apply liquid to one side of the membrane and apply physical force to the membrane surface by hydrostatic pressure, pressurization, vacuum suction, etc. When the liquid is allowed to pass through the membrane filter, it is used for the purpose of separating the substance into a substance larger and smaller than the pores of the membrane filter. Alternatively, nucleic acid, protein, etc. can be transferred, adsorbed and fixed to the membrane and stained directly, such as Southern blotting, Northern blotting, Western blotting, and dot plotting, or with a specific binding substance labeled with RI or a fluorescent substance. It is used for the purpose of searching and testing for the presence or absence of interactions.

  Further, it is also used as a base material for electrophoresis in which a sample solution is supplied to one end of a membrane filter filled with an electrolyte solution, a voltage is applied to both ends of the membrane, and a substance is separated by interaction with the base material. Recently, an in vitro diagnostic agent that measures the presence or absence of substances in body fluids by directly donating urine, blood, etc., using immunochromatography, which takes advantage of both the chromatographic and blotting base materials of membrane filters. Has been developed and is widely used in applications such as pregnancy testing, fecal occult blood, and drug testing.

  In recent years, with the improvement of home medical care and regional medical care such as clinics and clinics, as well as the early diagnosis and the increase in urgent clinical tests, even if you are not a clinical laboratory specialist, There is a growing demand for analyzers that can perform high-accuracy measurement simply and quickly. For this reason, highly reliable measurement can be performed in a short time without complicated operations. Small analyzers for POCT (POINT OF CARE TESTING) are in the spotlight.

  POCT is a general term for tests performed in “close to patients” such as practitioners, specialists' examination rooms, wards, and clinics for outpatients. It is a method that has been attracting attention as a great help in improving the quality of medical care by performing various treatments and performing treatment processes and even prognostic monitoring. Compared to inspections in the central laboratory, such small analyzers can reduce the cost of sample transport, equipment, and unnecessary inspections, reducing total inspection costs. The POCT market is said to be possible, and is rapidly expanding in the US, where hospital management rationalization is progressing, and is expected to become a growth market in Japan and other countries.

  In the field of clinical examination, for the purpose of fractionating serum proteins, a large number of electrophoretic supports comprising a microporous membrane mainly composed of a polymer such as cellulose acetate are used. The characteristics of this electrophoresis are that it has a high resolution for charge differences, requires a very small amount of sample, has a very short migration time, and is suitable for protein analysis in various body fluids such as serum, urine, and cerebrospinal fluid. ing. However, although it is cellulose acetate membrane electrophoresis that has many advantages in terms of ease of use, careful handling is necessary for handling the cellulose acetate membrane used for electrophoresis, and electrophoresis that can be used more easily There has been a need for a substrate.

  In addition, dry analytical elements such as immunochromatographic sensors do not require the preparation of reagents, and only by simple operations such as dropping a liquid sample such as blood or urine to be measured onto the analytical element. Since it is possible to analyze the substance to be inspected in the liquid sample, and it is very useful for analyzing the substance to be inspected in the liquid sample simply and quickly, it is practically used today as a representative of POCT. It has become. Further, the market demands higher accuracy in analysis in addition to being able to measure anyone anytime and anywhere.

  This immunochromatographic sensor has a reaction part in which a reagent that specifically reacts with an analyte is immobilized on a porous carrier such as nitrocellulose or glass fiber filter paper, and specifically reacts with the analyte and the immobilized reagent. A labeling reagent holding part labeled with the reagent to be provided is provided, and a sample addition part for adding a liquid sample is provided on the labeling reagent holding part or upstream of the labeling reagent holding part in the reagent developing direction.

  The measurement principle is that a liquid sample is first added to the sample addition section. The added liquid sample develops on the porous carrier, reaches the labeling reagent holding site, develops further downstream, passes through the reaction layer, and the liquid sample develops on the porous carrier downstream. Here, when the analyte is contained in the liquid sample, the analyte first undergoes a specific binding reaction with the labeling reagent, develops to the reaction part, and further reaches the reaction part and then reacts specifically with the labeling reagent. The analyzed object causes a specific reaction with the immobilized reagent in the reaction part, and a color reaction according to the concentration of the analyzed object can be confirmed. Using this principle, many pregnancy diagnostic agents, cancer marker diagnostic agents, myocardial marker diagnostic agents and the like are commercially available.

  Many immunochromatographic sensors were mainly qualitative, but recently, as seen in myocardial marker diagnostics, there are products that read the color reaction at the reaction site and measure it with a quantitative device. It is appearing.

  However, when performing quantitative measurement with an immunochromatographic sensor, the information is only information obtained by optically reading the degree of coloration in the reaction part, and the amount of information is too small to carry out more accurate quantitative measurement. It was extremely difficult.

  Therefore, various methods have been taken for the purpose of incorporating electrochemical techniques as means for increasing the number of information. For example, in Patent Document 1 (Japanese Patent Application Laid-Open No. 11-76765), a porous film in which a porous region partitioned by providing an impermeable portion in the porous film is formed is reported. A porous film is described in which electrodes and wiring portions are formed on a cellulose membrane using a gold paste ink by screen printing.

  Further, in Patent Document 2 (International Publication No. 01/004614 pamphlet), in a lateral flow type immunochromatography sensor, pure cellulose chromatography paper is used as a printing substrate, and silver / silver chloride paste ink is used on the substrate. A biosensor composed of a chromatography matrix part having electrodes and an electrode part has been reported.

  In any of the above-described methods, the electrode portion is formed by screen printing on a porous film formed in advance. However, when screen printing is performed, most of the printing paste is adjusted to a paste by an organic solvent. In many porous membranes, when an organic solvent permeates, the porous shape is denatured, and a phenomenon occurs in which the porous shape is destroyed. Therefore, when forming electrodes by screen printing, the porous shape is destroyed and the role of the original porous film cannot be fulfilled, so when printing electrodes on the porous film, the choice of the solvent is very It was important.

  In addition, when the electrodes are formed by screen printing, there is a problem that the measurement electrode area varies due to the bleeding of the paste at the time of printing, and the response characteristics vary. This problem occurs very frequently in the case of Patent Documents 1 and 2 in which an electrode is formed on a porous membrane. Therefore, it is important to be solved in improving quantitative accuracy in a biosensor using these carriers. It was an issue. Moreover, since the yield deteriorates when only uniform items are selected, there is a problem that the cost is further increased.

  In order to solve these problems, the measurement of the electrode area after printing on the porous film is an indispensable inspection process. By incorporating this process, the measurement accuracy of the sensor is slightly improved, but by adding the inspection process The increase in man-hours is added to the cost of the sensor. Therefore, even if the sensor performance is improved, there is a problem that it is impossible to provide an inexpensive sensor. There has been a strong demand for the provision of inexpensive materials that can be subjected to dynamic analysis.

  Patent Document 3 (Japanese Patent Laid-Open No. 8-75748) reports an electrode type immune reaction analyzer using a flow-through method. This method has a structure in which an antibody-insolubilized porous film is placed on an electrode prepared by screen printing on PET without adhering, and a specific binding reaction occurs in the porous film. Quantitative measurement is performed by measuring the amount of a substance obtained from an enzyme-labeled antibody with an electrode.

  By adopting this configuration, the electrodes and the porous film are produced separately compared to the case where the electrodes are directly formed on the porous film as in Patent Documents 1 and 2, and printing is directly performed on the porous film. Since the paste is not applied, it is possible not only to prevent the porous film from being destroyed, but also to expand the choice of the printing paste for electrode printing, and the frequency of occurrence of variations in the electrode area due to printing bleeding is very low. Thus, by providing a more uniform material, the possibility of performance improvement is expected as compared with Patent Documents 1 and 2.

  However, since highly accurate measurement is impossible unless the antibody-insolubilized porous membrane and electrode prepared separately are accurately laminated, position control is required to assemble these members into sensors accurately. Manufacturing process, introduction of assembly equipment, and confirmation of assembly variation must be introduced. Although these problems can be solved moderately by these introductions, it has been impossible to provide an inexpensive sensor due to the increase in the number of parts and the cost increase due to the introduction of the inspection process. .

Materials having a porous membrane and an electrically conductive layer that can improve both measurement accuracy and reduce costs as described above, and that can realize electrochemical analysis have been widely studied.
JP-A-11-76765 International Publication No. 01/004614 Pamphlet JP-A-8-75748

  As described above, those disclosed in Patent Documents 1 to 3 are materials or sensors intended for electrochemical detection and quantification in a biosensor based on an immune reaction. By using these materials or sensors, it was devised to enable electrochemical measurement of immunosensors, which had previously only been able to perform qualitative determinations, to perform quantitative measurements with higher accuracy and accuracy. It was an invention.

  However, in any of the inventions, it is very difficult to provide a material having a uniform property, and even if the devised material or sensor shape is used, more accurate assembly accuracy is required, or the property of the material. Any number of inspection processes for confirmation must be introduced, and more stable materials must be provided, and it is far from accurate and highly accurate quantitative measurement. It has been extremely difficult to provide an inexpensive sensor that can be measured.

  In order to solve these problems and realize more accurate and highly accurate quantitative measurement, an inexpensive high-accuracy material that does not require built-in accuracy and has a small variation in the electrically conductive layer and electrode area is used. Providing was an important and essential element.

  The present invention has been made in view of the above problems, and as a result of intensive studies, in a biosensor based on an immune reaction, while improving measurement accuracy, it does not impair the convenience found in conventional immunosensors. The purpose is to provide a low-cost and high-accuracy material that can realize electrochemical analysis in order to perform more accurate and high-precision measurement that anyone can measure anytime, anywhere.

  In order to solve the above problems, the porous carrier according to claim 1 of the present invention is formed so that the porous film is fixed on the electrically conductive layer formed on one surface of the insulating substrate. It is characterized by this.

  A porous carrier according to a second aspect of the present invention is the porous carrier according to the first aspect, wherein the electrically conductive layer is formed on the entire upper surface or a part of the upper surface of the insulating substrate. The porous film is formed on the entire or part of the upper surface of the conductive layer.

  A porous carrier according to a third aspect of the present invention is the porous carrier according to the first or second aspect, wherein the electrically conductive layer is formed by vapor deposition or sputtering. Is.

  A porous carrier according to a fourth aspect of the present invention is the porous carrier according to the first or second aspect, wherein the electrically conductive layer is formed by printing.

  A porous carrier according to a fifth aspect of the present invention is the porous carrier according to the first to fourth aspects, wherein one or more electrodes are formed in the electrically conductive layer. It is.

  A porous carrier according to a sixth aspect of the present invention is the porous carrier according to any one of the first to fifth aspects, wherein the electrically conductive layer is divided by slits.

  A porous carrier according to a seventh aspect of the present invention is the porous carrier according to the sixth aspect, wherein the slit is formed by laser processing an electrically conductive layer. .

  A porous carrier according to an eighth aspect of the present invention is the porous carrier according to any one of the first to seventh aspects, wherein the porous film is a water-absorbing carrier or a water-absorbing carrier. To do.

  The porous carrier according to claim 9 of the present invention is the porous carrier according to any one of claims 1 to 8, wherein the porous membrane comprises nitrocellulose, regenerated cellulose, polyvinylidene fluoride (PVDF), nylon, polycarbonate, It is one type of pure film selected from the group consisting of cellulose acetate or a mixed film of two or more types.

  A porous carrier according to a tenth aspect of the present invention is the porous carrier according to any one of the first to ninth aspects, wherein the insulating substrate is made of a transparent liquid-impermeable material.

  A dry analytical element according to an eleventh aspect of the present invention uses the porous carrier according to any one of the first to tenth aspects.

  According to a twelfth aspect of the present invention, there is provided an immunological test substrate using the porous carrier according to any one of the first to tenth aspects.

  A support for electrophoresis according to claim 13 of the present invention is characterized in that the porous carrier according to any one of claims 1 to 10 is used.

  An immunochromatographic assay substrate according to a fourteenth aspect of the present invention is characterized by using the porous carrier according to any one of the first to tenth aspects.

  According to a fifteenth aspect of the present invention, there is provided a porous carrier manufacturing method, wherein an electrically conductive layer is formed on an upper surface of an insulating substrate, and a porous film is fixed on the upper surface of the previously formed electrically conductive layer. It is characterized by this.

  A porous carrier manufacturing method according to claim 16 of the present invention is the porous carrier manufacturing method according to claim 15, wherein an electrically conductive layer is formed on the upper surface of the insulating substrate by any method, Using the conductive layer-forming insulating substrate as a support, obtaining the step of applying a material for forming a porous film on the upper surface of the electrically conductive layer by any method to form the porous film To do.

  The porous carrier manufacturing method according to claim 17 of the present invention is the porous carrier manufacturing method according to claim 15, wherein an electrically conductive layer is formed on the upper surface of the insulating substrate by any method. A porous carrier is formed by laminating a porous film formed in advance on the upper surface of the electrically conductive layer of the formed insulating substrate.

  A porous carrier manufacturing method according to claim 18 of the present invention is the porous carrier manufacturing method according to claim 17, wherein the electrically conductive layer-forming insulating substrate and the porous film are bonded together. It is characterized by forming.

  A porous carrier manufacturing method according to a nineteenth aspect of the present invention is the porous carrier manufacturing method according to the eighteenth aspect, wherein the porous film and the electrically conductive layer-forming insulating substrate or the insulating substrate and the electric material are electrically connected. The conductive layer is bonded to the conductive layer by thermocompression bonding.

  A porous carrier manufacturing method according to a twentieth aspect of the present invention is the porous carrier manufacturing method according to the eighteenth aspect, wherein an ultrasonic wave is applied while the members are in contact with each other, The electrically conductive layer-forming insulating substrate or the insulating substrate and the electrically conductive layer are bonded together.

  The porous carrier manufacturing method according to claim 21 of the present invention is the porous carrier manufacturing method according to claim 15 to 20, wherein the electrically conductive layer is formed by vapor deposition or sputtering. Is.

  A porous carrier manufacturing method according to claim 22 of the present invention is the porous carrier manufacturing method according to claims 15 to 20, wherein the electrically conductive layer is formed by printing.

  A porous carrier manufacturing method according to claim 23 of the present invention is the porous carrier manufacturing method according to claim 15 to 22, wherein one or more electrodes are formed on the electrically conductive layer, and then the porous membrane is formed. It is characterized by forming.

  A porous carrier manufacturing method according to claim 24 of the present invention is the porous carrier manufacturing method according to claims 15 to 22, wherein the electrically conductive layer is divided by a slit and then a porous film is formed. It is characterized by.

  A porous carrier manufacturing method according to claim 25 of the present invention is the porous carrier manufacturing method according to claim 24, wherein the slit is formed by processing an electrically conductive layer with a laser. To do.

  A porous carrier manufacturing method according to claim 26 of the present invention is the porous carrier manufacturing method according to claim 15 to 22, wherein the insulating substrate is a transparent or translucent liquid-impermeable material. It is what.

  A porous carrier manufacturing method according to claim 27 of the present invention is the porous carrier manufacturing method according to claim 26, wherein after the electrically conductive layer and the porous film are formed, from the insulating substrate side. The electric conductive layer is divided by slits.

  A porous carrier manufacturing method according to a twenty-eighth aspect of the present invention is the porous carrier manufacturing method according to the twenty-seventh aspect, wherein the slit is processed by a YAG laser.

  Since the porous carrier according to claim 1 of the present invention is formed so as to fix the porous film on the electrically conductive layer formed on the insulating substrate, the electrically conductive layer prepared individually and Since it is possible to provide an integrated material without the need to combine with porous membranes, measurements that make use of the characteristics of porous membranes and measurements that make use of the characteristics of electrically conductive layers can be performed simultaneously. It becomes possible. In addition, since combination processing is not necessary, not only high-precision measurement can be performed, but also entry into a field where measurement has conventionally been difficult is possible. For example, an electrode pattern is formed with an electrically conductive layer on an insulating substrate, and a porous film is formed on a part of the electrically conductive layer, thereby providing an electrode-type biosensor equipped with a blood cell filter. It becomes possible. Further, it is possible to provide a chromatographic sensor capable of electrochemical analysis by utilizing the fact that the porous membrane support is an electrically conductive layer. Since the electrically conductive layer and the porous film are configured as the same carrier, there is no need to assemble the electrically conductive layer and the porous film with high precision when a sensor is produced using this carrier. By realizing this material, it is possible not only to reduce the cost by reducing the sensor manufacturing process, but also to use an electrode type with a blood cell filter, which has been considered to be impossible to measure with high accuracy because it is impossible to achieve high-precision assembly. It has the effect of enabling the realization of biosensors and electrode-type chromatographic sensors.

  Note that the adhesion shown here is an insulating substrate and an electrically conductive layer and an electrically conductive and porous film or an insulating substrate and a porous film that are closely stacked without moving individually. It is in a state where it cannot be separated or peeled into three layers without applying a strong force such as scraping, and each member can be stored in a stacked case or left using a clip or pin etc. It is completely different from what you are doing.

  The insulating substrate is made of an insulating liquid-impermeable material, and may be transparent, translucent, or opaque. The material is not only synthetic resin materials such as PET, ABS, polystyrene, and polyvinyl chloride. It is possible to use an insulating liquid-impermeable material such as glass.

  Further, the porous membrane refers to a thin film having a shape in which micropores are continuously connected, preferably a polymer filter generally called a membrane filter, and having a shape in which pores having the same pore diameter are continuously connected. What you are doing is good. The polymer material constituting the porous membrane is not particularly limited, and examples thereof include nitrocellulose, regenerated cellulose, polyvinylidene fluoride (PVDF), nylon, polycarbonate, and cellulose acetate. The formation of these porous membranes is not particularly limited, and there is no problem even if they are produced by any of the methods used for producing commercially available membrane filters. The preferable pore diameter of the porous membrane that can be used in the present invention is 0.02 to 20 μM as the average pore diameter, and the particularly preferable average pore diameter is 0.1 to 10 μM. In addition to the average pore size, the membrane filter used in the present invention preferably has a uniform pore size. Further, the preferred film thickness of the porous film is 10 to 150 μM, but these are only examples, and there is no problem even if the pore diameter or film thickness is outside this range. Moreover, although it is not a membrane filter, it does not matter at all to form filter agents, such as a cellulose filter paper and a glass filter.

  The electrically conductive layer refers to a layer made of a substance capable of transferring electric charge, for example, an electrically conductive substance such as noble metal such as gold or palladium, or carbon, and they are deposited or sputtered. Or it is the metal film formed by adhere | attaching metal foil, or it is formed by arbitrary construction methods, such as screen printing, using various paste ink. These are only examples, and there is no problem even if the conductive layer is made of a material or method other than those described above.

  A porous carrier according to a second aspect of the present invention is the porous carrier according to the first aspect, wherein the electrically conductive layer is formed on the whole or a part of the upper surface of an insulating substrate. Since the porous film is formed on the whole or part of the upper surface of the conductive layer, the shapes of the electrically conductive layer and the porous film can be freely changed according to the use of the porous carrier. By doing so, it has the effect of being able to develop applications in a wide range of fields.

  For example, if you want to create an electrode-type biosensor equipped with a blood cell filter, you only need to form a porous membrane only at the site that corresponds to the sample introduction part. If you want to create a biosensor that has a measurement site on which protein is immobilized, What is necessary is just to form a porous film only in the location corresponded to a site | part. Further, an electrophoretic membrane can be realized by forming an electroconductive layer on the lower surface of the porous membrane to be chromatographed so as to correspond to a positive electrode and a negative electrode. Note that these are only examples, and there is no problem with the usage methods other than those described above, and the formation patterns of the electrically conductive layer and the porous film can be freely combined depending on the purpose. If conventional materials are used, assembly accuracy is important in order to place a porous membrane at a specific location on the electrode. The assembly process, jigs for more accurate assembly, introduction of equipment, and inspection after assembly Although the process has been considered indispensable, if the porous carrier according to claim 2 of the present invention is used, it is not necessary at all, and therefore it is possible to provide a material that can realize further cost reduction.

  The porous carrier according to claim 3 of the present invention is the porous carrier according to claim 1 or 2, wherein the electrically conductive layer is formed by vapor deposition or sputtering. Since a uniform thin film with high film thickness accuracy can be formed and a highly accurate electroconductive layer can be formed, the use of this porous material has the effect of greatly improving measurement accuracy. The vapor deposition shown here refers to heating and evaporating a metal or a compound in a vacuum and applying the vapor to the surface of the object in a thin film. Sputtering blows an inert gas onto the metal and blows it away. It is preferable to form an electrically conductive layer with an apparatus capable of realizing these methods by attaching the molecules to the surface of the object.

  A porous carrier according to a fourth aspect of the present invention is the porous carrier according to the first or second aspect, wherein the electrically conductive layer is formed by printing. By forming a single layer, or printing and laminating silver paste, carbon paste, etc., there is an effect that the electric conductivity can be controlled by a combination of materials of the electric conductive layer. The printing shown here has no problem with any method such as thick film printing such as screen printing, non-impact printing such as inkjet, and patterning by roll coating such as gravure printing.

  A porous carrier according to a fifth aspect of the present invention is the porous carrier according to any one of the first to fourth aspects, wherein one or more electrodes are formed on the electrically conductive layer. Since an electrode has already been formed under the membrane, it is not necessary to add electrode printing in consideration of the structural deformation of the membrane in post-processing, and it is possible to provide a porous membrane that can use electrical control It becomes. By using this porous carrier, there is an effect that a biosensor capable of an electric control function and electrochemical analysis can be realized.

  A porous carrier according to a sixth aspect of the present invention is the porous carrier according to any one of the first to fifth aspects, wherein the electrically conductive layer is divided by a slit, and is generated during printing. There is no variation in the electrode area due to bleeding or dripping of the printing paste, and the area of each electrode can be defined with higher accuracy, and a porous carrier with more uniform performance can be provided. Have The slits shown here indicate that the electrically conductive layer is divided. As a method of dividing, any method such as cutting a part of the electrically conductive layer with a laser or a jig having a sharp tip or a printing pattern is used. Because it is formed by, there is no problem.

  The porous carrier according to claim 7 of the present invention is the porous carrier according to claim 6, wherein the slit is formed by laser processing of an electrically conductive layer. When slits can be processed and electrodes are formed, the area of each electrode can be controlled with high accuracy, and a porous carrier having an electrically conductive layer that can assume a minute electrode can be provided. . The laser processing shown here indicates that processing is performed using a gas laser, a solid laser, a semiconductor laser, a dye laser, or the like. Specific examples of these lasers include CO2 laser, YAG laser, and excimer laser. However, there is no problem even if other than these lasers are used. These lasers may be selected in accordance with the processing. For example, when the insulating substrate is transparent, if the YAG laser is used, slit processing can be performed not only from the electrically conductive layer side but also from the insulating substrate side. It is possible to use a YAG laser when the metal film having a thickness of several μM is an electrically conductive layer. For a metal film having a thickness of several NM, any of CO2, YAG, and excimer laser can be used. It may be used. The combination and selection of lasers are not limited to these.

  The porous carrier according to claim 8 of the present invention is the porous carrier according to any one of claims 1 to 7, wherein the porous film is a water-absorbing carrier or a carrier that has been treated to absorb water. The porous carrier is rich in liquid permeability, and if this is used, there is an effect that it is possible to provide a material capable of realizing a biosensor capable of analyzing a liquid sample.

  The porous carrier according to claim 9 of the present invention is the porous carrier according to any one of claims 1 to 8, wherein the porous membrane comprises nitrocellulose, regenerated cellulose, polyvinylidene fluoride (PVDF), nylon, polycarbonate, Since it is one type of pure membrane selected from the group consisting of cellulose acetate or a mixed membrane of two or more types, it is possible to provide a porous membrane useful as a substrate for hybridization or immunoassay. Has an effect. In addition, since a porous film having micropores is formed and the pore diameter can be freely selected, a material that can be used as a membrane-holding electrically conductive layer having a function as a filter can be provided.

  A porous carrier according to a tenth aspect of the present invention is the porous carrier according to any one of the first to ninth aspects, wherein the insulating substrate is made of a transparent liquid-impermeable material. In the case of using, since it is not possible to confirm slits or electrodes of the electrically conductive layer after forming the porous film, it is necessary to perform some positioning before forming the porous film. If it exists, it has the effect that it becomes possible to confirm the position of a slit or an electrode from the insulating substrate side. In the case of using this porous carrier, more accurate positioning is possible, and higher accuracy can be achieved.

  Since the dry analytical element according to the eleventh aspect of the present invention uses the porous carrier according to any one of the first to tenth aspects, the electrically conductive layer and the porous film are integrated. The number of assembly steps can be reduced, and the electroconductive layer is very homogeneous, so the accuracy in electrical analysis is very good, and it is possible to realize a dry analytical element with high accuracy and low cost. It has the effect to become.

  Since the immunological test substrate according to claim 12 of the present invention uses the porous carrier according to any one of claims 1 to 10, not only the electrochemical analysis is realized, but also the electrical conductivity. Since the layer and the porous membrane are integrated, the number of assembly steps can be reduced, and since the electrically conductive layer is very homogeneous, the accuracy in the electrical analysis is very good, high accuracy and low cost It has the effect of making it possible to realize a simple immunological test substrate.

  Since the support for electrophoresis according to claim 13 of the present invention uses the porous carrier according to any one of claims 1 to 10, the support for electrophoresis is not only stronger and less likely to be damaged. In addition, since the electrically conductive layer and the porous membrane are integrated, an electrophoretic support that is very easy to handle and that is simply connected to the electrode is provided. It has the effect that becomes possible.

  Since the immunochromatographic assay substrate according to claim 14 of the present invention uses the porous carrier according to any one of claims 1 to 10, not only realizes electrochemical analysis in immunochromatographic assay, Since the electrically conductive layer and the porous membrane are integrated, the number of sensor assembly steps can be reduced, and the sensor can be manufactured more easily, and the electrically conductive layer is very homogeneous. This has the effect of realizing an immunochromatographic assay substrate with extremely high accuracy in analysis and high accuracy and low cost.

  According to a fifteenth aspect of the present invention, there is provided a porous carrier manufacturing method in which an electrically conductive layer is formed on an upper surface of an insulating substrate, and a porous film is fixed on the upper surface of the previously formed electrically conductive layer. Therefore, there is no need to prepare and assemble individual objects, and there is no need for the bonding accuracy or the assembly accuracy, and it is possible to produce a more homogeneous porous carrier.

  Note that the adhesion shown here is an insulating substrate and an electrically conductive layer and an electrically conductive and porous film or an insulating substrate and a porous film that are closely stacked without moving individually. It is in a state where it cannot be separated or peeled into three layers without applying a strong force such as scraping, and each member can be stored in a stacked case or left using a clip or pin etc. It is completely different from what you are doing.

  The insulating substrate is made of an insulating liquid-impermeable material, and may be transparent, translucent, or opaque. The material is not only synthetic resin materials such as PET, ABS, polystyrene, and polyvinyl chloride. It is possible to use an insulating liquid-impermeable material such as glass.

  Further, the porous membrane refers to a thin film having a shape in which micropores are continuously connected, preferably a polymer filter generally called a membrane filter, and having a shape in which pores having the same pore diameter are continuously connected. What you are doing is good. The polymer material constituting the porous membrane is not particularly limited, and examples thereof include nitrocellulose, regenerated cellulose, polyvinylidene fluoride (PVDF), nylon, polycarbonate, and cellulose acetate. The formation of these porous membranes is not particularly limited, and there is no problem even if they are produced by any of the methods used for producing commercially available membrane filters. The preferable pore diameter of the porous membrane that can be used in the present invention is 0.02 to 20 μM as the average pore diameter, and the particularly preferable average pore diameter is 0.1 to 10 μM. In addition to the average pore size, the membrane filter used in the present invention preferably has a uniform pore size. Further, the preferred film thickness of the porous film is 10 to 150 μM, but these are only examples, and there is no problem even if the pore diameter or film thickness is outside this range. Moreover, although it is not a membrane filter, it does not matter at all to form filter agents, such as a cellulose filter paper and a glass filter.

  The electrically conductive layer refers to a layer made of a substance capable of transferring electric charge, for example, an electrically conductive substance such as noble metal such as gold or palladium, or carbon, and they are deposited or sputtered. Or it is the metal film formed by adhere | attaching metal foil, or it is formed using arbitrary construction methods, such as screen printing, using various paste ink. These are only examples, and there is no problem even if the conductive layer is made of a material or method other than those described above.

  A porous carrier manufacturing method according to claim 16 of the present invention is the porous carrier manufacturing method according to claim 15, wherein an electrically conductive layer is formed on the upper surface of the insulating substrate by any method, Since the conductive layer forming insulating substrate is used as a support, the porous film is formed by obtaining a step of applying a material for forming the porous film on the upper surface of the electrically conductive layer by an arbitrary method. There is no need to bond individual members, and it is not necessary to pursue the bonding accuracy, and it is possible to produce a lower cost and homogeneous porous carrier.

  The porous carrier manufacturing method according to claim 17 of the present invention is the porous carrier manufacturing method according to claim 15, wherein an electrically conductive layer is formed on the upper surface of the insulating substrate by any method. Since the porous carrier is formed by laminating a porous film formed in advance on the upper surface of the electrically conductive layer of the formed insulating substrate, it is impossible to directly produce the porous carrier on the electrically conductive layer. Even in the porous membrane, it is possible to realize a porous carrier integrated with the electrically conductive layer, and it is possible to produce a more homogeneous porous carrier.

  A porous carrier manufacturing method according to claim 18 of the present invention is the porous carrier manufacturing method according to claim 17, wherein the electrically conductive layer-forming insulating substrate and the porous film are bonded together. As a result, a porous carrier integrated with the electrically conductive layer can be realized even in a porous film that cannot be directly formed on the electrically conductive layer, and the carrier is more electrically conductive. This has the effect of producing a more homogeneous porous carrier in which the layer and the porous membrane are in close contact.

  A porous carrier manufacturing method according to a nineteenth aspect of the present invention is the porous carrier manufacturing method according to the eighteenth aspect, wherein the porous film and the electrically conductive layer-forming insulating substrate or the insulating substrate and the electric material are electrically connected. Since the conductive layer is bonded by thermocompression bonding, it is possible to make the porous layer that does not like organic solvents adhere more firmly to the conductive layer when bonding the conductive layer and the porous film. Thus, the carrier has the effect of making it possible to produce a more homogeneous porous carrier in which the electrically conductive layer and the porous membrane are tightly adhered. In addition, the thermocompression bonding shown here is a method in which an electrically conductive layer is placed on an insulating substrate coated with a paste that melts at a constant temperature, and the paste is melted at a temperature equal to or higher than the melting point of the paste. Each material is placed in a temperature environment of room temperature or higher, not limited to these methods, such as laminating a conductive substrate, or crimping at a temperature higher than the melting point of the polymer resin constituting the porous film or insulating substrate. , Indicates pasting each other.

  A porous carrier manufacturing method according to a twentieth aspect of the present invention is the porous carrier manufacturing method according to the eighteenth aspect, wherein an ultrasonic wave is applied while the members are in contact with each other, Since the electrically conductive layer-forming insulating substrate or the insulating substrate and the electrically conductive layer are bonded together, a porous film that does not like organic solvents or heat is more firmly adhered to the electrically conductive layer. The carrier has the effect of making it possible to produce a more homogeneous porous carrier in which the electrically conductive layer and the porous membrane are in close contact with each other. In addition, the bonding by applying the ultrasonic wave shown here applies ultrasonic waves such as applying ultrasonic waves in a state where the respective members are in contact with each other to generate frictional heat and melting and bonding. The method of bonding together is shown.

  Since the porous carrier manufacturing method according to claim 21 of the present invention is the porous carrier manufacturing method according to claims 15 to 20, the electrically conductive layer is formed by vapor deposition or sputtering. A uniform thin film with high film thickness accuracy can be formed, and a highly accurate electroconductive layer can be formed. Therefore, it is possible to manufacture a homogeneous porous carrier capable of measuring with higher accuracy. It has the effect to become. The vapor deposition shown here refers to heating and evaporating a metal or a compound in a vacuum and applying the vapor to the surface of the object in a thin film. Sputtering blows an inert gas onto the metal and blows it away. It is preferable to form an electrically conductive layer with an apparatus capable of realizing these methods by attaching the molecules to the surface of the object.

  The porous carrier manufacturing method according to claim 22 of the present invention is the porous carrier manufacturing method according to claims 15 to 20, wherein the electrically conductive layer is formed by printing. Thus, it is possible to produce a porous carrier adjusted to have an electric conductivity more suitable for the application. The printing shown here has no problem with any method such as thick film printing such as screen printing, non-impact printing such as inkjet, and patterning by roll coating such as gravure printing.

  A porous carrier manufacturing method according to claim 23 of the present invention is the porous carrier manufacturing method according to claim 15 to 22, wherein one or more electrodes are formed on the electrically conductive layer, and then the porous membrane is formed. Therefore, there is no need to produce an electrode in the subsequent process on the porous membrane, and a porous membrane having an electrically conductive layer can be easily produced, producing a more uniform porous carrier. It has the effect that becomes possible.

  A porous carrier manufacturing method according to claim 24 of the present invention is the porous carrier manufacturing method according to claims 15 to 22, wherein the electrically conductive layer is divided by a slit and then a porous film is formed. Therefore, it is possible to divide the electrically conductive layer with high accuracy, and there is no variation in the electrode area due to bleeding or dripping of the printing paste that has occurred in the case of printing. This has the effect of making it possible to produce a more homogeneous porous carrier that can define the area with higher accuracy. In addition, the slit shown here shows dividing | segmenting an electroconductive layer, As a method of dividing | segmenting, a part of an electroconductive layer is shaved with a laser, a jig | tool with a sharp tip, or a printing pattern etc. Since it is formed by an arbitrary method, there is no problem.

  The porous carrier manufacturing method according to claim 25 of the present invention is the porous carrier manufacturing method according to claim 24, wherein the slit is formed by processing the electrically conductive layer with a laser. The effect of being able to process a highly accurate slit that can make the electrode area uniform with high precision in the case of forming an electrode, etc., and producing a more homogeneous porous carrier that enables highly accurate measurement Have In addition, finer electrode processing is possible. The laser processing shown here indicates that processing is performed using a gas laser, a solid laser, a semiconductor laser, a dye laser, or the like. Specific examples of these lasers include CO2 laser, YAG laser, and excimer laser. However, there is no problem even if other than these lasers are used. These lasers may be selected in accordance with the processing. For example, when the insulating substrate is transparent, if the YAG laser is used, slit processing can be performed not only from the electrically conductive layer side but also from the insulating substrate side. It is possible to use a YAG laser when the metal film having a thickness of several μM is an electrically conductive layer. For a metal film having a thickness of several NM, any of CO2, YAG, and excimer laser is used. May be. The combination and selection of lasers are not limited to these.

  The porous carrier production method according to claim 26 of the present invention is the porous carrier production method according to claim 15 to 22, wherein the insulating substrate is a transparent or translucent liquid-impermeable material. Therefore, when an opaque insulating substrate is used, slits and electrodes of the electrically conductive layer cannot be confirmed after the porous film is formed. Therefore, it is necessary to perform some positioning before forming the porous film. However, if it is transparent or translucent, the position of the slit and the electrode can be confirmed from the insulating substrate side. In the case of using this porous carrier, more accurate positioning is possible, and higher accuracy can be achieved.

  A porous carrier manufacturing method according to claim 27 of the present invention is the porous carrier manufacturing method according to claim 26, wherein after the electrically conductive layer and the porous film are formed, from the insulating substrate side. Since the electrically conductive layer is divided by the slit, there is an effect that the slit can be more accurately divided into an arbitrary position. In addition, when slit division is performed after the formation of the electrically conductive layer, when a slit different from the formed slit division pattern is formed, it is necessary to newly prepare a porous carrier, but the porous carrier is formed. Since arbitrary slit division can be performed later, it is possible to process a necessary amount of the porous carrier as required.

  The porous carrier manufacturing method according to claim 28 of the present invention is the porous carrier manufacturing method according to claim 26, wherein the slit is processed by a YAG laser, and therefore, the transparent carrier is made transparent by taking advantage of the characteristics of the YAG laser. Alternatively, there is an effect that the electrically conductive layer can be divided into slits without damaging or modifying the translucent insulating substrate and the porous film.

  As described above, according to the porous carrier and the porous carrier production method of the present invention, when producing a biosensor that requires a porous membrane or requires electrochemical analysis, an electrode is attached to the porous membrane. There is no need to fabricate or assemble the electrode and porous membrane, and since the porous membrane has an electrically conductive layer, it not only reduces the man-hours of the sensor production process, but also pursues assembly accuracy. This eliminates the need for accuracy inspection processes such as assembly accuracy and assembly jigs, so that it is possible to drastically reduce sensor production costs and to provide materials that can produce low-cost and high-precision biosensors. By using this porous carrier, it is possible to realize a more accurate and highly accurate measurement method that can be easily and quickly measured by anyone at any time and anywhere.

  Hereinafter, embodiments of the porous carrier of the present invention will be described in detail with reference to the drawings. In addition, embodiment shown here is an example to the last, Comprising: It is not necessarily limited to this embodiment.

(Embodiment 1)
FIG. 1 is a perspective view showing the configuration of the porous carrier according to the first embodiment, and FIG. 2 is a perspective view showing an example of the method for producing the porous carrier according to the first embodiment. In the porous carrier of the first embodiment, 1 is an insulating substrate made of polyethylene terephthalate or the like, 2 is a conductive material mainly composed of carbon or carbon formed on the entire upper surface of the insulating substrate 1, or Reference numeral 3 denotes an electrically conductive layer made of a noble metal such as gold or palladium, and 3 denotes a porous film made of nitrocellulose or the like formed on the entire upper surface of the electrically conductive layer 2.

  The insulating substrate 1 is made of an insulating liquid-impermeable material such as a PET film, and may be transparent, translucent, or opaque. The material is a synthetic resin such as ABS, polystyrene, or polyvinyl chloride. In addition to the material, it is possible to use an insulating liquid-impermeable material such as glass.

  In addition, the electrically conductive layer 2 is a metal layer formed by vapor deposition, sputtering, or adhesion of a metal foil, or a conductive layer made of paste ink such as silver or carbon formed by using any method such as screen printing. It consists of

  The porous membrane used in the present invention is a polymer filter generally called a membrane filter, and has a shape in which pores having the same pore diameter are continuously connected. The polymer material constituting the porous membrane is not particularly limited, and examples thereof include nitrocellulose, regenerated cellulose, polyvinylidene fluoride (PVDF), nylon, polycarbonate, and cellulose acetate. The formation of these porous membranes is not particularly limited, and may be produced by any of the methods used for producing commercially available membrane filters. The preferable pore diameter of the porous membrane that can be used in the present invention is 0.02 to 20 μM as the average pore diameter, and the particularly preferable average pore diameter is 0.1 to 10 μM. In addition to the average pore size, the membrane filter used in the present invention preferably has a uniform pore size. Moreover, although the suitable film thickness of a porous film is 10-150 micromol, there is no problem at the film thickness outside this range. Further, although it is not a membrane filter, it is not a problem to form a filter medium such as cellulose filter paper or glass filter.

  An example of this porous carrier manufacturing method will be described with reference to FIG. First, a palladium thin film that could become the electrically conductive layer 2 was formed on the entire upper surface of the insulating substrate 1 using a sputtering apparatus. Next, a porous thin film 3 having a porous film 3 having a film thickness of 150 μM and a pore diameter of 5 μM was applied by uniformly applying and drying a nitrocellulose resin dissolved in an organic solvent on the entire upper surface of the palladium thin film using the insulating substrate on which the palladium thin film was formed as a support. A quality carrier was formed. In the porous carrier formed in this way, the porous film and the electrically conductive layer are firmly adhered, and the insulating substrate, the porous film, and the electrically conductive layer exist as an integral solid body.

  Next, FIG. 3 is a perspective view showing a porous carrier in which a porous film is formed on a part of the electrically conductive layer, and FIGS. 4 and 5 are electrical conductors formed on a part of the upper surface of the insulating substrate. It is an example of the perspective view which shows the porous support | carrier provided with the porous film formed in the porous layer and the whole surface. FIG. 1 shows a porous carrier comprising an electrically conductive layer formed on the entire upper surface of an insulating substrate and a porous film formed on the entire upper surface of the electrically conductive layer, and FIG. However, as shown in FIG. 3, a state where a porous film is formed at an arbitrary position which is a part of the upper surface of the electrically conductive layer 2 formed on the entire upper surface of the insulating substrate, FIG. As shown in FIG. 5, there is no problem even if an electrically conductive layer is provided on a part of the upper surface of the insulating substrate and a porous film is formed on the entire upper surface of the electrically conductive layer.

  For example, the porous carrier shown in FIG. 4 is porous on the entire upper surface of the electrically conductive layer 2 in which slits 5 are formed on the palladium thin film (metal film 4) using a laser on the entire upper surface of the insulating substrate 1 using a sputtering apparatus. In FIG. 5, the insulating substrate 1 has an electrically conductive layer 2 provided with a printing electrode 6 having a silver electrode pattern printed on a part of its upper surface by screen printing. The porous film is formed on the entire upper surface of the conductive layer 2. An example of a method for producing the porous carrier of FIG. 4 is shown in FIG. 6, and an example of a method of producing the porous carrier of FIG. 5 is shown in FIG.

  In the method of FIG. 6, a palladium thin film (metal film 4) having a thickness of 10 NM was formed on the entire upper surface of a PET sheet having a thickness of 100 μM, which is the insulating substrate 1, using a sputtering apparatus. Next, the slit 5 was formed in the palladium thin film (metal film 4) surface using the YAG laser. Using the insulating substrate on which the electrically conductive layer 2 is formed as a support, a nitrocellulose resin dissolved in an organic solvent is applied to the entire surface of the electrically conductive layer 2 made of a slit-formed palladium thin film formed on the upper surface of the insulating substrate. Drying to form a porous carrier provided with the porous membrane 3 having a film thickness of 150 μM and a pore diameter of 5 μM.

  Further, in the method of FIG. 7, the silver paste was printed on the PET sheet having a thickness of 100 μM, which is the insulating substrate 1, using the screen printing method, and the printed electrode 6 was formed. Applying and drying a nitrocellulose resin dissolved in an organic solvent on the entire surface of the electroconductive layer 2 made of a silver printed electrode, using the insulating substrate provided with the electroconductive layer 2 made of the printed electrode 6 as a support, A porous carrier provided with the porous membrane 3 having a film thickness of 150 μM and a pore diameter of 5 μM was formed.

  FIG. 8 is an example of a process flow diagram of the porous carrier manufacturing method according to the first embodiment. As shown in the above-described production flow example 1 represented by FIG. 4 and FIG. 5, the porous carrier is produced by forming an electrically conductive layer directly on the upper surface of the insulating substrate, and forming an electrically conductive layer insulating property. A porous film is formed separately as in a method of forming a porous carrier by directly forming a porous film on the upper surface of the substrate, or a manufacturing flow example 2, and an electrically conductive layer is directly formed on the upper surface of the insulating substrate. A method for producing a porous carrier by adhering to the upper surface of the electrically conductive layer of the formed electrically conductive layer forming insulating substrate by an arbitrary method, and producing an electrically conductive layer separately as in Production Flow Example 3 An electrically conductive layer is formed on the upper surface of the insulating substrate by an arbitrary method to produce an electrically conductive layer forming insulating substrate, and a porous film is directly formed on the electrically conductive layer surface of the electrically conductive layer forming insulating substrate. Method for producing porous carrier, electrically conductive layer as in production flow example 4 Porous membranes are prepared individually, and an electrically conductive layer is applied to the upper surface of the insulating substrate by an arbitrary method, an electrically conductive layer forming insulating substrate is formed, and an electrically conductive layer surface is formed by an arbitrary method. Examples include, but are not limited to, a method of attaching a porous film and preparing a porous carrier.

  In the first embodiment, the porous carrier preparation method including the formation of a metal film by sputtering, the production of a printed electrode by screen printing, the formation of a slit using a YAG laser, etc. has been described, but this is not restrictive. There is no problem even if a metal film forming method, a printing method, or a slit forming method other than the above method is used.

  In addition, the arbitrary sticking method is a method using an adhesive, thermocompression bonding, or sticking by applying ultrasonic waves, but is not limited thereto, and a method of sticking individual members If so, there is no problem.

(Embodiment 2)
FIG. 9 is a conceptual diagram of an electrode-type biosensor using the porous carrier of the second embodiment. In FIG. 9, 11 is a space forming part for sucking a certain amount of liquid sample, 12 is an electrode type biosensor, and 16 is a processing line when processing a porous carrier as a sensor. The porous carrier of Embodiment 2 includes an electrically conductive layer 2 on the upper surface of an insulating substrate 1, and the electrically conductive layer 2 forms a slit 5 by laser processing, and the slit 5 forms an electrically conductive layer. 2 is configured to include a measurement electrode and a counter electrode. In addition, a porous film 3 is formed on a part of the upper surface of the electrically conductive layer 2 and a portion corresponding to the bottom surface of the space forming portion 11 provided for adding a certain amount of sample.

  The porous membrane 3 is impregnated with a reagent for measuring an enzyme reaction. For example, when the electrode-type biosensor measures glucose concentration in blood, the porous membrane 3 contains an electron mediator composed of potassium ferricyanide and an enzyme for degrading glucose, glucose dehydrogenase (GDH). ing. A certain amount of blood is sucked into the space forming part 11 from the opening 12, and the glucose in the sucked blood reacts with GDH in the porous film. Electron mediator mediates enzyme reaction and electrode reaction to cause the movement of electrons as a signal. Electrons generated by both reactions move through the electron mediator and measure the amount of electrons as a current value. Is used to measure the current value through the electrically conductive layer, whereby the glucose concentration can be measured.

  Here, since the porous membrane 3 is provided in advance under the space forming portion and on the upper surface of the electrically conductive layer, it is not necessary to install the porous membrane on the electrode at the time of assembling the sensor. An inspection process for confirming the installation position of the porous membrane is not necessary. Furthermore, there is no need to worry about deterioration in accuracy due to the displacement of the combination position. Also, by using this porous carrier, blood cell components in blood can be filtered and prevented from adhering to the electrode, and blood cells do not adhere to the electrically conductive layer, so that electron transfer in cell membranes such as erythrocytes Therefore, it is possible to provide a highly accurate and highly sensitive electrode-type biosensor that is not affected by blood cell components that have hindered accurate measurement.

  In addition, although glucose measurement was mentioned here as an example of a biosensor, and it demonstrated using potassium ferricyanide, GDH, etc. as an example of the reagent of a sensor structure, for example, electron mediators, such as ferrocene, enzymes, such as glucose oxidase, etc. Including other mediators, enzymes or other reagents, there is no problem, not only for the measurement of blood glucose concentration, but also for blood samples such as cholesterol and lactic acid, liquid samples other than blood, and analytes other than glucose There is no problem even if the measurement is intended. The configuration of the electrode-type biosensor shown in FIG. 9 is only an example, and the configuration of the biosensor is not limited to this. The processing line 16 shows a shape where it can be said that the porous carrier has been processed into a grid pattern. However, the processing line 16 is not limited to this, and there is no problem even if it is a processing line punched out with a mold or the like. .

(Embodiment 3)
FIG. 10 is a conceptual diagram of an immunosensor using the porous carrier of the third embodiment. In FIG. 10, 14 is a labeled reagent impregnated portion that is held so that a labeled reagent labeled with a specific protein that specifically reacts with an analyte in the liquid sample can be dissolved, and 15 is an analyzed object in the liquid sample. A specific protein immobilization part in which a specific protein that specifically reacts with a substance is immobilized on the porous membrane 3, and 7 is a configuration example of an immunosensor comprising the porous carrier of the present invention and the above-described components 16. Indicates a processing line when processing a porous carrier as a sensor.

  The porous carrier of the third embodiment includes an electrically conductive layer 2 on the upper surface of the insulating substrate 1, and the electrically conductive layer 2 forms a slit 5 by laser processing, and the slit 5 is electrically conductive. Layer 2 is configured to include a measurement electrode and a counter electrode. A porous film 3 made of nitrocellulose is formed on the entire upper surface of the electrically conductive layer 2.

  The porous membrane 3 is provided with a specific protein immobilization section 14 and a labeling reagent impregnation section 15 capable of performing a specific reaction with an analysis object. When the liquid sample sucked from the space forming part 11 reaches the labeling reagent impregnation part 14, the labeling reagent is dissolved. Both the liquid sample and the labeling reagent perform a specific reaction with the analyte in the liquid sample while developing the porous film 3, and reach the specific protein immobilization unit 15. In the specific protein immobilization unit 15, a specific reaction is performed between the analyte in the liquid sample, the labeling reagent, and the immobilized protein, and some color reaction occurs. Excess liquid sample develops downstream of the specific protein immobilization unit 15 in the chromatographic direction. The color reaction at the specific protein immobilization unit 15 is optically read using a measuring device, and the analyte concentration in the liquid sample is qualitatively or quantitatively measured.

  Here, since the porous film 3 is provided on the upper surface of the electrically conductive layer in advance, it is not necessary to install the porous film on the electrode at the time of assembling the sensor. An inspection process for confirming the position is not necessary. Furthermore, there is no need to worry about deterioration in accuracy due to the displacement of the combination position. Further, by using this porous carrier, the electroconductive layer 2 is provided on the lower surface of the porous membrane, which is a chromatographic carrier, so that an enzyme is used as a labeling reagent and a specific reaction in a specific protein immobilization part. It is also possible to measure the current value resulting from the above using a measuring device. For example, when a peroxidase labeling reagent is used as a labeling reagent, phenylenediamine is used as a redox reagent, and hydrogen peroxide is used as a substrate, the color reaction derived from the redox reaction is optically measured, and the electrons derived from the enzyme reaction are electrochemically measured. By measuring automatically, it becomes possible to realize more accurate and accurate quantitative measurement, and provide highly accurate and very inexpensive measurement.

  Here, a method is used in which a porous film is formed on the electrically conductive layer provided with the slit 5, but the electrically conductive layer is formed on the entire upper surface of the insulating substrate, and the entire upper surface of the electrically conductive layer is porous. There is no problem even if a method of forming a slit by irradiating a porous carrier provided with a film with a laser from the insulating substrate side. In addition, the specific reaction indicates an antigen-antibody reaction, and an antigen-antibody reaction such as a sandwich reaction using a different specific protein that exhibits an antigen-antibody reaction against an analyte as a labeling reagent and an immobilizing reagent, or a competitive reaction is used Indicates any reaction. Peroxidase and phenylenediamine are only examples, and there is no problem using any enzyme, substrate, and coloring dye.

  The configuration of the electrode-type biosensor shown in FIG. 10 is only an example, and the configuration of the biosensor is not limited to this. The processing line 16 shows a shape where it can be said that the porous carrier has been processed into a grid pattern. However, the processing line 16 is not limited to this, and there is no problem even if it is a processing line punched out with a mold or the like. .

(Embodiment 4)
FIG. 11 is an electrophoretic support using the porous carrier according to the fourth embodiment, and FIG. 12 is a schematic diagram illustrating an electrophoretic method. In FIG. 11, 17 represents an electrophoretic support, 9 in FIG. 12 represents a negative pole, and 10 represents a positive pole.

  The porous carrier according to the fourth embodiment includes an electrically conductive layer 2 formed by printing on a part of the insulating substrate 1, and the porous film 3 made of cellulose acetate on the entire upper surface of the electrically conductive layer 2. Is configured to form. After the protein is applied to the porous carrier and impregnated in, for example, 0.5 M Tris-1.5 M glycine (PH 8.8) electrophoresis buffer, the electrically conductive layer 2 provided at both ends of the porous carrier. One is connected to the positive electrode and the other is connected to the negative electrode, allowing a constant current to flow. When the porous carrier after electrophoresis is reacted with the protein staining solution, an electrophoresis pattern corresponding to the molecular weight of the protein is obtained on the porous carrier 8.

  In this example, 0.5 M Tris-1.5 M glycine (PH 8.8) was used as the electrophoresis buffer. However, the present invention is not limited to this, and there is no problem even if other buffers are used. There is no problem even if a porous membrane is used.

  The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

(Preparation of porous carrier)
A palladium thin film having a thickness of 10 NM was formed on a transparent PET (polyethylene terephthalate) sheet having a thickness of 100 μM, a width of 1 M, and a length of 50 M using a sputtering apparatus. Using a palladium thin film PET sheet as a support, a nitrocellulose resin dissolved in an organic solvent is applied to the upper surface of the palladium thin film layer side, and a nitrocellulose membrane having a pore diameter of 5 μM and a thickness of 150 μM is formed into a roll having a width of 1 M and a length of 30 M. Formed (FIG. 2).

(Slit formation of porous carrier)
From the transparent insulating substrate side of the porous carrier, slitting is completed by dividing the palladium thin film layer with slits by laser trimming using a YAG laser to form measurement electrodes, counter electrodes, lead portions, etc. A porous carrier was produced.

(Quantification of CRP concentration in whole blood)
A) Adjustment of immunosensor FIG. 13 is a schematic view showing a procedure for producing the immunosensor of this example. This figure mainly shows a method for assembling each member, and details of the manufacturing method of this embodiment will be described below based on this manufacturing procedure.

  First, a strip having a width of 50 MM and a length of 30 CM was cut from the slit-formed porous carrier roll.

  An anti-CRP antibody A solution was prepared by diluting with a phosphate buffer solution to adjust the concentration. This antibody solution was applied to a position of 25 MM from the end of the width 50 MM direction on the porous film of the slit-formed porous carrier produced above using a solution discharge device. Thereby, the immobilized antibody part I15 which is a specific protein immobilization part was obtained on the porous membrane which consists of nitrocellulose. Next, in the same manner, the anti-CRP antibody B solution was applied to a part 2 mm away from the sample addition part on the downstream side. After drying this porous membrane, it was immersed in a TRIS-HCL buffer solution containing 1% skim milk and gently shaken for 30 minutes. After 30 minutes, the membrane was moved to the TRIS-HCL buffer solution tank, gently shaken for 10 minutes, and then gently shaken for another 10 minutes in another TRIS-HCL buffer solution tank to wash the membrane. Next, after immersing in a 0.1 M TRIS-HCL (PH 8.9) buffer solution containing 0.05% sucrose monolaurate and gently shaking for 10 minutes, the membrane was removed from the liquid bath and allowed to cool at room temperature. Dried. As a result, a specific protein immobilization part 15 consisting of an immobilized antibody part I and an immobilized antibody part II was obtained on the nitrocellulose membrane.

  The enzyme-labeled anti-CRP antibody labeled with peroxidase 1: 1 was dissolved in 0.1 M phosphate buffer PH 6.0 containing 1% sucrose. To this enzyme-labeled antibody solution, P-phenylenediamine was added as an electron mediator to a final concentration of 15 MM to prepare a labeling reagent solution containing the electron mediator. A liquid separated from the immobilization part I and the immobilization part II on the dry porous membrane coated with the immobilized anti-CRP antibody A and the immobilized anti-CRP antibody B by setting the labeling reagent solution in a solution discharge device In order from the sample addition start direction, coating was performed so that the labeling reagent, the immobilization part I, and the immobilization part II were in a positional relationship, and then the porous membrane was vacuum freeze-dried. As a result, a porous carrier provided with a specific protein immobilization part and an enzyme-labeled antibody labeling reagent impregnation part was obtained.

  Next, a transparent tape was affixed from the specific protein immobilization part and the labeling reagent impregnation part 14 of the porous carrier containing the prepared labeling reagent to the terminal part. Then, it cut | disconnected to the sensor size of width 5.0MM and length 50.0MM using CO2 laser. After cutting, a space forming part 11 prepared by laminating transparent PET having a thickness of 100 μM is pasted on the starting end part where the transparent tape is not pasted, and a fine space (width 5.0 MM × length) having an opening 12 is pasted. 7.0MM × 0.3MM in height) 6 was formed. Thus, the immunosensor 7 was manufactured.

B) Preparation of liquid sample CRP concentrations of 0MG / DL, 0.3MG / DL, 5MG / DL are added to human blood (hematocrit 40%) with heparin added as an anticoagulant by adding a CRP solution of known concentration. The blood was adjusted.

C) Measurement of reaction on sensor A sample solution to which hydrogen peroxide was added to 1.0 M with respect to 50 μL of whole blood containing CRP prepared in B) above was spotted from opening 12. Five minutes after arrival, the absorbance at a wavelength of 490 NM in the specific protein immobilization part and the current value at an applied voltage of 150 MV were measured. The result is shown in FIG. In FIG. 13, the horizontal axis represents the CRP concentration of the added blood, and the vertical axis represents the absorbance and current value obtained from the specific protein immobilization part. Referring to this figure, both the absorbance and current value in the immobilization part I tend to increase according to the CRP concentration in the blood. On the other hand, the absorbance and the current value in the immobilization part II change as the CRP concentration increases. It should be noted here that the absorbance is 0 at 0 MG / DL and 0.3 MG / DL, whereas the current value is slightly increased at 0.3 MG / DL. From this result, while it is not possible to measure the reaction in the immobilized part II with a concentration of 0.3 MG / DL in terms of absorbance, it is possible to sufficiently measure the difference in the three concentrations with respect to the current value. Then, the detection sensitivity is increased, that is, by using the porous carrier of the present invention, electrochemical analysis in an immunochromatographic sensor can be performed, and high-sensitivity measurement can be performed as compared with conventional optical detection. I can ask.

  Further, according to the production procedure of FIG. 14, since the porous carrier has the porous membrane and the electrode part fixed to each other, the porous film in which the antibody is immobilized in advance and the electrode sheet on which the electrode part is formed are provided. It is possible to omit the assembly process. Furthermore, when assembling both the porous membrane and the electrode part, in addition to the position control for specific protein immobilization in the porous film and the position control for electrode part preparation, the assembly of the porous film and the electrode part in advance. Sometimes the position of the specific protein immobilization part and the position control of the electrode part are very important, but if the porous carrier in this example is used, the position is controlled only with respect to the setting of the porous carrier, and the work starting point is set. If it is constant, the position control of all parts can be performed precisely, the manufacturing procedure can be simplified, and the inspection process for position control can be omitted.

  As for the assembly position of this porous membrane and electrode part, if the position of specific protein immobilization and the electrode part for reading the reaction are shifted, naturally the amount of current derived from the reaction will decrease, so the sensitivity will only decrease. In addition, if the deviation width is different among the individual sensors, the degree of reduction in sensitivity is also different, so the measurement accuracy is also deteriorated. However, if the above porous carrier is used, no displacement occurs, so that the current is accurately captured by the electrode section and sufficient sensitivity is obtained, and the displacement width between individual sensors is not different, so high accuracy Measurement can be realized.

  In the above examples, a transparent PET sheet was used as the insulating substrate, and a process flow in which slit formation was performed by laser processing after formation of the porous carrier was used. However, there is no problem even if laser processing is performed before formation of the porous film. Not limited to transparent, a translucent or opaque PET sheet may be used.

  In addition, the measurement of CRP concentration in whole blood has been described, but not limited thereto, antibodies, immunoglobulins, hormones, enzymes in any test such as blood test, urine test, water test, stool test, soil analysis, food analysis, etc. And any liquids, including proteins and protein derivatives such as peptides, infectious substances such as bacteria, viruses, fungi, mycoplasma, parasites and their products and components, drugs and tumor markers such as drugs of abuse and abuse Measurement with any analyte in the sample is possible. Further, the configuration of the sensor is not limited to this, and there is no problem as long as the immune reaction and the electrochemical analysis can be performed.

  According to the porous carrier and the porous carrier production method of the present invention, when producing a biosensor that requires a porous membrane or requires electrochemical analysis, an electrode is produced on the porous membrane, There is no need to assemble the electrode and porous membrane, and since the porous membrane has an electrically conductive layer, it is not only possible to reduce the man-hour of the sensor manufacturing process but also to pursue assembly accuracy. This eliminates the need for accuracy inspection processes such as assembly accuracy, assembly jigs, and the like, so that it is possible to drastically reduce sensor production costs and to provide materials that can produce low-cost and high-precision biosensors. . On the other hand, if this porous carrier is used, it is possible to realize a more accurate, high-accuracy, and high-sensitivity measurement method that anyone can easily and quickly measure anytime, anywhere.

1 is a perspective view of a porous carrier according to a first embodiment of the present invention. The perspective view which shows the preparation methods of the porous carrier by Embodiment 1 of this invention The perspective view which shows the porous support | carrier which formed the porous film in 1 part of the electrically conductive layer concerning Embodiment 1 of this invention The perspective view which shows the porous support | carrier provided with the electroconductive layer formed in a part of upper surface of the example insulating board | substrate concerning Embodiment 1 of this invention, and the porous film | membrane formed in the whole surface. The perspective view which shows the porous support | carrier provided with the electroconductive layer formed in a part of upper surface of the example insulating board | substrate concerning Embodiment 1 of this invention, and the porous film | membrane formed in the whole surface. The perspective view which shows the preparation methods of the porous support | carrier shown in FIG. The perspective view which shows the preparation methods of the porous support | carrier shown in FIG. Process flowchart which shows an example of the production process of the porous support | carrier concerning Embodiment 1 of this invention. The perspective view which shows the electrode-type biosensor using the porous support | carrier concerning Embodiment 2 of this invention. A perspective view showing an immunosensor using a porous carrier according to a third embodiment of the present invention. The perspective view which shows the porous support | carrier concerning Embodiment 4 of this invention. The perspective view which shows the usage example of the porous support | carrier concerning Embodiment 4 of this invention. Explanatory drawing which shows the preparation procedure of the immunosensor in the Example of this invention. The figure which shows the measurement result using the immunosensor in the Example of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Insulating substrate 2 Electrically conductive layer 3 Porous film 4 Metal film 5 Slit 6 Print electrode 7 Immunosensor 8 Electrophoresis support 9 Negative electrode 10 Positive electrode 11 Space formation part 12 Opening part 13 Electrode type biosensor 14 Label Reagent impregnation part 15 Specific protein immobilization part 16 Processing line 17 Transparent tape

Claims (28)

A porous carrier comprising a porous film fixed on an electrically conductive layer formed on at least one surface of an insulating substrate. The porous carrier according to claim 1, wherein
The electrically conductive layer is formed on the whole or part of the upper surface of the insulating substrate,
A porous carrier comprising the porous film formed on the entire upper surface or a part of the upper surface of the electrically conductive layer. The porous carrier according to claim 1 or 2,
The porous carrier, wherein the electrically conductive layer is formed by vapor deposition or sputtering. The porous carrier according to claim 1 or 2,
The porous carrier is characterized in that the electrically conductive layer is formed by printing. The porous carrier according to any one of claims 1 to 4,
A porous carrier comprising one or more electrodes formed on the electrically conductive layer. The porous carrier according to any one of claims 1 to 5,
The porous carrier, wherein the electrically conductive layer is divided by slits. The porous carrier according to claim 6, wherein
The porous carrier, wherein the slit is an electrically conductive layer formed by laser processing. The porous carrier according to any one of claims 1 to 7,
A porous carrier, wherein the porous membrane is a water-absorbing carrier or a carrier treated so as to have water absorption. The porous carrier according to any one of claims 1 to 8,
The porous film is one type of pure film selected from the group consisting of nitrocellulose, regenerated cellulose, polyvinylidene fluoride (PVDF), nylon, polycarbonate, and cellulose acetate, or a mixed film of two or more types. A porous carrier. The porous carrier according to any one of claims 1 to 9,
A porous carrier, wherein the insulating substrate is made of a transparent liquid-impermeable material. A dry analytical element using the porous carrier according to any one of claims 1 to 10. An immunological test substrate using the porous carrier according to any one of claims 1 to 10. 11. A support for electrophoresis, wherein the porous carrier according to claim 1 is used. An immunochromatographic assay substrate using the porous carrier according to any one of claims 1 to 10. A method for producing a porous carrier, characterized in that a porous film is fixed on an electrically conductive layer formed on at least one surface of an insulating substrate. In the porous carrier manufacturing method according to claim 15,
An electrically conductive layer is formed by an arbitrary method on the upper surface of the insulating substrate, and a material for forming a porous film on the electrically conductive layer is formed using the electrically conductive layer forming insulating substrate as a support. A method for producing a porous carrier, comprising obtaining the step of coating by a method to form the porous film. In the porous carrier manufacturing method according to claim 15,
An electrically conductive layer is formed on the upper surface of the insulative substrate by an arbitrary method. An electrically conductive layer is formed by laminating a pre-formed porous film on the upper surface of the electrically conductive layer of the insulating substrate. A method for producing a porous carrier, comprising forming a carrier. The method for producing a porous carrier according to claim 17,
A method for producing a porous carrier comprising forming a porous carrier by laminating an electrically conductive layer-forming insulating substrate and a porous film. The method for producing a porous carrier according to claim 18,
A method for producing a porous carrier, wherein the porous film and the electrically conductive layer-forming insulating substrate, or the insulating substrate and the electrically conductive layer are bonded together by thermocompression bonding. The method for producing a porous carrier according to claim 18,
The porous film and the electrically conductive layer forming insulating substrate, or the insulating substrate and the electrically conductive layer are bonded together by applying ultrasonic waves in a state where the members are in contact with each other. A method for producing a porous carrier. The method for producing a porous carrier according to claim 15 to 20, wherein
The method for producing a porous carrier, wherein the electrically conductive layer is formed by vapor deposition or sputtering. The method for producing a porous carrier according to claim 15 to 20, wherein
The method for producing a porous carrier, wherein the electrically conductive layer is formed by printing. The method for producing a porous carrier according to claims 15 to 22,
A method for producing a porous carrier, comprising forming a porous film after forming one or more electrodes on the electrically conductive layer. The method for producing a porous carrier according to claims 15 to 22,
A porous carrier production method comprising forming a porous film after dividing the electrically conductive layer by slits. The method for producing a porous carrier according to claim 24,
The said slit forms an electroconductive layer by laser processing, The porous carrier manufacturing method characterized by the above-mentioned. The method for producing a porous carrier according to claims 15 to 22,
A method for producing a porous carrier, wherein the insulating substrate is a transparent or translucent liquid-impermeable material. In the porous carrier manufacturing method according to claim 26,
After forming the said electroconductive layer and a porous membrane, the said electroconductive layer is divided | segmented by a slit from the said insulating substrate side, The porous carrier manufacturing method characterized by the above-mentioned. The method for producing a porous carrier according to claim 27,
A method for producing a porous carrier, wherein the slit is processed with a YAG laser.

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