A biosensor and its manufacturing method

Abstract

The embodiment of the present application provides a biosensor and a manufacturing method thereof, which belongs to the field of electrochemical analysis technology, and includes a substrate; a power supply electrode and a plurality of working electrodes arranged on the substrate; the substrate has a reaction area, the reaction area includes a first area and a plurality of independent second areas, each second area is connected to the first area, the signal end of the power supply electrode is located in the first area, and the signal ends of the plurality of working electrodes are respectively located in different second areas; a hydrophilic layer, the hydrophilic layer is arranged on the power supply electrode and the plurality of working electrodes, and the hydrophilic layer covers the reaction area; a hydrophobic layer, the hydrophobic layer is arranged on the substrate, and the hydrophobic layer surrounds the hydrophilic layer. Through the biosensor and the manufacturing method thereof provided by the embodiment of the present application, the mutual interference between the working electrodes can be reduced, and the accuracy of the detection results can be improved.

Description

Technical Field

[0001] The embodiments of the present application relate to the field of electrochemical analysis technology, and more specifically, to a biosensor and a method for manufacturing the same. Background Art

[0002] Electrochemical enzyme sensors are enzyme biosensors, and their basic structural unit is composed of a substance recognition element (immobilized enzyme membrane) and a signal converter (substrate electrode). When an enzymatic reaction occurs on the enzyme membrane, the electroactive substances produced are responded to by the substrate electrode. The function of the substrate electrode is to convert chemical signals into electrical signals for detection.

[0003] At present, biosensors usually include multiple working electrodes, on which different specific enzymes are fixed respectively. Different specific enzymes can catalyze the corresponding analytes in the liquid to react and generate products to make the working electrodes work. Therefore, biosensors can detect multiple components in the liquid to be detected.

[0004] However, since the products of reactions of different analytes catalyzed by different specific enzymes may be the same, and all working electrodes are located on the same substrate, when the distances between different working electrodes are very close, mutual interference may occur between the different working electrodes, resulting in errors in the detection results of the components in the liquid. Summary of the invention

[0005] The embodiments of the present application provide a biosensor and a method for manufacturing the same, which are intended to reduce mutual interference between working electrodes and improve the accuracy of detection results.

[0006] A first aspect of an embodiment of the present application provides a biosensor, comprising a substrate;

[0007] A power supply electrode and a plurality of working electrodes are disposed on the substrate;

[0008] The substrate has a reaction area, the reaction area includes a first area and a plurality of mutually independent second areas, each of the second areas is connected to the first area, the signal end of the power electrode is located in the first area, and the signal ends of the plurality of working electrodes are respectively located in different second areas;

[0009] A hydrophilic layer, wherein the hydrophilic layer is disposed on the power electrode and the plurality of working electrodes, the hydrophilic layer covers the reaction area, and an edge of the hydrophilic layer extends to an edge of the substrate;

[0010] The hydrophobic layer is disposed on the substrate and surrounds the hydrophilic layer.

[0011] Optionally, the material of the hydrophilic layer includes: silane containing carboxyl groups.

[0012] Optionally, the material of the hydrophobic layer includes: silane containing fluorine or chlorine, and insulating paste.

[0013] Optionally, a protective layer is provided, wherein the protective layer covers the hydrophilic layer and the hydrophobic layer, and two sides of the protective layer are connected to the substrate.

[0014] Optionally, the plurality of working electrodes are located on one side or both sides of the power electrode;

[0015] Alternatively, the plurality of working electrodes are evenly spaced apart along the circumference of the power electrode.

[0016] Optionally, the power supply electrode includes: an active electrode.

[0017] Optionally, the power supply electrode includes: a counter electrode and a reference electrode.

[0018] A second aspect of the embodiments of the present application provides a method for manufacturing a biosensor, the method comprising: providing a substrate;

[0019] forming a power electrode and a plurality of working electrodes on the substrate by a printing process and drying the electrodes, wherein the signal end of the power electrode and the signal ends of the plurality of working electrodes are located in a reaction area of ​​the substrate;

[0020] Covering the power electrode and the leads of the plurality of working electrodes with an insulating paste to form an insulating layer, and drying the layers;

[0021] Printing reagent enzymes on the plurality of working electrodes by a printing process and drying them;

[0022] forming a hydrophilic layer on the substrate, wherein the hydrophilic layer covers the reaction area;

[0023] A hydrophobic layer is formed on the substrate, wherein the hydrophobic layer surrounds the hydrophilic layer.

[0024] Optionally, forming the hydrophilic layer on the substrate comprises:

[0025] The substrate is treated with oxygen plasma to generate OH groups, thereby forming the hydrophilic layer.

[0026] Beneficial effects:

[0027] The present application provides a biosensor and a method for manufacturing the same, wherein a power electrode and a plurality of working electrodes are arranged on a substrate, and the power electrode is located in a first region on the substrate, and each working electrode is located in a different second region, and then a hydrophilic layer and a hydrophobic layer are arranged on the substrate, and the hydrophilic layer covers the reaction region; when the biosensor is used for detection, the liquid to be detected is allowed to enter the hydrophilic layer in the first region, and due to the hydrophilicity of the hydrophilic layer, the liquid to be detected will diffuse on the hydrophilic layer and eventually diffuse to each second region, at which time the reagent enzyme on the working electrode will react with the corresponding component in the liquid to be detected; the hydrophobic layer can also prevent the liquid to be detected from diffusing to other locations.

[0028] Since each working electrode is located in a different second area, and the second areas are independent of each other, and a hydrophobic layer is arranged between the second areas, the product generated by the reagent enzyme reaction cannot move directly to the position of other working electrodes, thereby reducing the mutual interference between different working electrodes and improving the accuracy of biosensor detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the description of the embodiments of the present application will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0030] Figure 1 is a schematic structural diagram of a biosensor proposed in one embodiment of the present application;

[0031] Figure 2 is a schematic diagram of the side structure of a biosensor proposed in one embodiment of the present application;

[0032] Figure 3 is a schematic diagram of the structure of another biosensor proposed in one embodiment of the present application;

[0033] Figure 4 is a schematic diagram of the side structure of a biosensor proposed in one embodiment of the present application;

[0034] Figure 5 This is a schematic diagram of a protective layer structure of a biosensor proposed in one embodiment of the present application;

[0035] Figure 6 is a schematic structural diagram of a biosensor proposed in one embodiment of the present application;

[0036] Figure 7 is a schematic diagram of the structure of another biosensor proposed in one embodiment of the present application;

[0037] Figure 8 is a schematic diagram of the structure of another biosensor proposed in one embodiment of the present application;

[0038] Fig. 9 is a schematic diagram of the structure of another biosensor proposed in one embodiment of the present application;

[0039] Fig.10 is a schematic diagram of the structure of another biosensor proposed in one embodiment of the present application;

[0040] Fig.11 is a schematic diagram of the structure of another biosensor proposed in one embodiment of the present application;

[0041] Fig.12 is a flowchart of a method for manufacturing a biosensor according to an embodiment of the present application;

[0042] Fig.13 1 is a flowchart of a method for manufacturing a protective layer according to an embodiment of the present application.

[0043] Explanation of the accompanying drawings: 1. Substrate; 2. Power electrode; 21. Counter electrode; 22. Reference electrode; 3. Working electrode; 4. Hydrophilic layer; 5. Hydrophobic layer; 6. Protective layer; 61. Hydrophilic area; 62. Hydrophobic area; A. Reaction area; A1. First area; A2. Second area. DETAILED DESCRIPTION

[0044] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0045] Embodiment 1

[0046] Reference Figure 1 As shown, a biosensor disclosed in an embodiment of the present application includes a substrate 1 and electrodes arranged on the substrate 1. The biosensor disclosed in the embodiment of the present application is mainly used for detecting various components in liquids.

[0047] Specifically, the substrate 1 can be made of polyester resin or polyimide.

[0048] Reference Figure 1 As shown, the electrode includes a power supply electrode 2 and a plurality of working electrodes 3. The power supply electrode 2 serves to connect the circuit and stabilize the voltage. Both the power supply electrode 2 and the working electrode 3 have a signal terminal ( Figure 1 The circular area in the Figure 1The signal end of the working electrode 3 has a reagent enzyme on its surface, which can react with the corresponding analyte in the liquid to be detected, and produce a product through the reagent enzyme reaction. The product will undergo oxidation or reduction reaction on the working electrode 3 on the corresponding sensor to produce a change in the electrical signal; at the same time, the reagent enzymes on each working electrode 3 are different, so that a biosensor can be used to detect multiple components in the liquid to be detected, which can effectively improve the detection efficiency of the biosensor. Of course, a biosensor can also be used to detect a component, so that the sensors of multiple working electrodes 3 will produce changes in electrical signals, so that the detection results after calculation and statistics are more accurate.

[0049] Reference Figure 1 As shown, the substrate 1 has a reaction area A, and the signal end of the power electrode 2 and the signal ends of the plurality of working electrodes 3 are in the reaction area A on the substrate 1. The reaction area A includes a first area A1 and a plurality of mutually independent second areas A2, the number of the second areas A2 being the same as the number of the working electrodes 3, and each second area A2 is connected to the first area A1.

[0050] Specifically, the signal end of the power electrode 2 is located in the first area A1, and the signal ends of the plurality of working electrodes 3 are located in different second areas A2. It should be noted that the first area A1 and the second area A2 are not a specific space formed on the substrate 1, but a division of the substrate 1 itself, and the sizes of the first area A1 and the second area A2 are set according to actual needs. And the sizes of the second areas A2 can be the same or different.

[0051] Reference Figure 1 and Figure 2 As shown, the biosensor further includes a hydrophilic layer 4 and a hydrophobic layer 5. The hydrophilic layer 4 is disposed on the substrate 1 and completely covers the reaction area A, that is, the overall shape of the hydrophilic layer 4 is the same as the shape of the reaction area A, and the area of ​​the hydrophilic layer 4 also corresponds to the area included in the reaction area A. The hydrophilic layer 4 is hydrophilic, and after the liquid to be detected is dropped onto the hydrophilic layer 4, the liquid to be detected will diffuse on the hydrophilic layer 4.

[0052] The hydrophobic layer 5 is also disposed on the substrate 1, and the hydrophobic layer 5 surrounds the hydrophilic layer 4, that is, the periphery of the reaction area A is covered by the hydrophobic layer 5, and the hydrophobic layer 5 is hydrophobic and can prevent the flow of the liquid to be detected. Figure 3 As shown, the positions between each second area A2 are filled with the hydrophobic layer 5 .

[0053] When using the biosensor for detection, the liquid to be detected enters the hydrophilic layer 4 located in the first area A1 from the edge where the hydrophilic layer 4 overlaps with the substrate 1. Due to the hydrophilicity of the hydrophilic layer 4, the liquid to be detected will diffuse on the hydrophilic layer 4 and eventually diffuse to each second area A2. At this time, when the liquid to be detected diffuses to the position where the signal end of each working electrode 3 is located, the corresponding components in the liquid to be detected will react with different reagent enzymes on each working electrode 3. The products generated by the reagent enzyme and the corresponding components in the liquid to be detected will cause the working electrode 3 to generate an electrical signal, thereby realizing the detection of various components in the liquid to be detected. Of course, if there is no component in the liquid to be detected that reacts with a certain reagent enzyme, then the corresponding working electrode 3 will not generate an electrical signal.

[0054] Since each working electrode 3 is located in a different second area A2, and the second areas A2 are independent of each other, and a hydrophobic layer 5 is provided between the second areas A2, the product generated by the reagent enzyme reaction cannot directly move to the position of other working electrodes 3, thereby reducing the mutual interference between different working electrodes 3 and improving the accuracy of biosensor detection.

[0055] In one embodiment, the material of the hydrophilic layer 4 can be silane with a carboxyl group, and can be directly formed on the substrate 1 through a printing process.

[0056] In other embodiments, the surface of the substrate 1 may also be treated with oxygen plasma to generate OH groups on the surface of the substrate 1 , thereby forming a hydrophilic layer 4 on the surface of the substrate 1 .

[0057] In one embodiment, the material of the hydrophobic layer 5 can be selected from silane containing fluorine or chlorine, and can be directly formed on the substrate 1 through a printing process.

[0058] In other embodiments, the hydrophobic layer 5 may also be made of insulating paste, such as epoxy resin or polyvinylidene fluoride, and printed on the substrate 1 by screen printing.

[0059] In one embodiment, referring to Figure 3 and Figure 4 As shown, a protective layer 6 is disposed on the substrate 1, the protective layer 6 is parallel to the substrate 1, the spacing between the protective layer 6 and the substrate 1 is 0.2mm-3mm, and the protective layer 6 covers the hydrophilic layer 4 and the hydrophobic layer 5, and the protective layer 6 can play an isolating and protective role for the entire biosensor. The protective layer 6 can be made of PET material or PI material.

[0060] The connection between the protective layer 6 and the substrate 1 can be achieved by bonding with adhesive strips. When connecting, it is only necessary to set adhesive strips on both sides of the protective layer 6. The adhesive strips can be on the left and right sides of the protective layer, or on the upper and lower sides of the protective layer.

[0061] Reference Figure 5 As shown, the protective layer 6 has a hydrophilic area 61 with the same pattern as the hydrophilic layer, and a hydrophobic area 62 with the same pattern as the hydrophobic layer. The hydrophilic area 61 can be made of hydrophilic ink or slurry, such as silane with carboxyl groups, and is directly formed on the protective layer 6 by printing or screen printing. The surface of the protective layer 1 can also be treated with oxygen plasma to form the hydrophilic area 61 on the surface of the protective layer 6; the hydrophobic area 62 can be made of hydrophobic ink or insulating slurry, such as silane or epoxy resin containing fluorine or chlorine, and is directly formed on the substrate 1 by printing or screen printing.

[0062] The power supply electrode 2 includes two cases.

[0063] Reference Figure 6 As shown, in the first case, the power electrode 2 only includes the working electrode 21, and the working electrode 21 can simultaneously play the role of connecting the circuit and stabilizing the voltage. At this time, the material of the working electrode 21 is Ag / AgCl, in which the ratio of silver to silver chloride is 1:1, 6:4 or 7:3.

[0064] Reference Fig. 9 As shown, in the second case, the power electrode 2 includes a counter electrode 21 and a reference electrode 22, wherein the counter electrode 21 serves to connect the circuit, and the reference electrode 22 serves to stabilize the voltage. At this time, the material of the counter electrode 21 is the same as that of the working electrode 3.

[0065] The electrode material of the reference electrode 22 is Ag / AgCl, wherein the ratio of silver to silver chloride is 1:1, 6:4 or 7:3.

[0066] The plurality of working electrodes 3 are located on one side or both sides of the power electrode 2 ; or, the plurality of working electrodes 3 are evenly spaced along the circumference of the power electrode 2 .

[0067] In one embodiment, referring to Figure 6 As shown, the power electrode 2 only includes an active electrode 21; there are three working electrodes 3 in total, and the electrode detection ends of the three working electrodes 3 are on the same straight line on the substrate 1. Correspondingly, three sub-regions 121 are formed on the substrate 1, each sub-region 121 is in a long strip shape, and the three long strip sub-regions 121 are parallel to each other.

[0068] In one embodiment, referring to Figure 7 As shown, the power electrode 2 only includes an active electrode 21 ; a total of four working electrodes 3 are provided, and the four working electrodes 3 are respectively located on both sides of the counter electrode 21 .

[0069] In one embodiment, referring to Figure 8As shown, the power electrode 2 only includes the working electrode 21 ; there are four working electrodes 3 in total, and the four working electrodes 3 are evenly distributed along the circumference of the counter electrode 21 .

[0070] In one embodiment, referring to Fig. 9 As shown, the power electrode 2 includes a counter electrode 21 and a reference electrode 22, and there are three working electrodes 3 in total. The electrode detection ends of the three working electrodes 3 are on the same straight line on the substrate 1. Correspondingly, three sub-regions 121 are formed on the substrate 1, each sub-region 121 is in a long strip shape, and the three long strip sub-regions 121 are parallel to each other.

[0071] In one embodiment, referring to Fig.10 As shown, the power electrode 2 includes a counter electrode 21 and a reference electrode 22 ; a total of four working electrodes 3 are provided, and the four working electrodes 3 are respectively located on both sides of the counter electrode 21 and the reference electrode 22 .

[0072] In one embodiment, referring to Fig.11 As shown, the power electrode 2 includes a counter electrode 21 and a reference electrode 22 ; a total of four working electrodes 3 are provided, and the four working electrodes 3 are evenly distributed along the circumference of the counter electrode 21 and the reference electrode 22 .

[0073] Embodiment 2

[0074] Fig.12 A method for manufacturing a biosensor is shown, referring to Fig.12 As shown, an embodiment of the present application provides a method for manufacturing a biosensor, the method comprising:

[0075] Step 201: Provide a substrate 1.

[0076] Step 202: forming a power electrode 2 and a plurality of working electrodes 3 on a substrate 1 by a printing process and drying the same. The signal end of the power electrode 1 and the signal ends of the plurality of working electrodes 3 are located in a reaction area A of the substrate.

[0077] Specifically, the printing process can be screen printing. When printing the power electrode 2 and the working electrode 3, the arrangement and position of the power electrode 2 and the working electrode 3 can be set as needed, and hot air drying can be used during drying. The reaction area A includes a first area A1 and a plurality of mutually independent second areas A2. The signal end of the power electrode 2 is located in the first area A1, and the signal ends of the plurality of working electrodes 3 are respectively located in different second areas A2.

[0078] Step 203: Cover the power electrode 2 and the leads of the plurality of working electrodes 3 with an insulating paste to form an insulating layer 5, and dry the insulating layer.

[0079] Specifically, the insulating paste may be epoxy resin or polyvinylidene fluoride, and the insulating layer 5 plays a role in protecting the electrode.

[0080] Step 204 : forming a hydrophilic layer 4 on the substrate 1 , wherein the hydrophilic layer 4 covers the reaction area A.

[0081] Specifically, the hydrophilic layer 4 is made of silane containing a carboxyl group and is formed on the substrate 1 through a printing process, and the hydrophilic layer 4 completely covers the reaction area A of the substrate.

[0082] The hydrophilic layer 4 can also be formed on the surface of the substrate 1 by treating the surface of the substrate 1 with oxygen plasma, thereby generating OH groups on the surface of the substrate 1 .

[0083] Step 205 : forming a hydrophobic layer 5 on the substrate 1 , wherein the hydrophobic layer 5 surrounds the hydrophilic layer 4 .

[0084] Specifically, the hydrophobic layer 5 can be made of hydrophobic ink or insulating paste, such as silane or epoxy resin containing fluorine or chlorine, and is formed on the substrate 1 through a printing process or a screen printing process, and the hydrophobic layer 5 surrounds the hydrophilic layer 4, that is, the hydrophobic layer 5 surrounds the reaction area A.

[0085] Step 206: Print the reagent enzyme on the multiple working electrodes 3 by a printing process and dry them.

[0086] In one embodiment, referring to Fig.13 As shown, after forming the hydrophobic layer 5 on the substrate 1, the method further comprises:

[0087] Step 301 : Setting adhesive strips on both sides of the substrate 1 .

[0088] Specifically, the length of the rubber strip is 5 mm, and the rubber strips on both sides are parallel to each other.

[0089] Step 302: Cover the substrate 1 with a protective layer 6 , and connect the protective layer 6 to the substrate 1 via a rubber strip.

[0090] Specifically, the protective layer 6 is made of plastic material, such as PET film or PI film.

[0091] It should be noted that the various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0092] It should also be noted that, in this article, the orientation or position relationship indicated by the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc. is based on the orientation or position relationship shown in the drawings, which is only for the convenience of describing the present application and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application. In addition, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply that there is any such actual relationship or order between these entities or operations, nor can they be understood as indicating or implying relative importance. Moreover, the terms "include", "comprise" or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article or terminal device including a series of elements includes not only those elements, but also includes other elements not explicitly listed, or also includes elements inherent to such process, method, article or terminal device. Without more restrictions, an element defined by the phrase "comprising a ..." does not exclude the existence of other identical elements in the process, method, article or terminal device comprising the element.

[0093] The technical solution provided by the present application is described in detail above. The principle and implementation method of the present application are described in this article using specific examples. The description of the above embodiments is only used to help understand the present application, and the content of this specification should not be understood as limiting the present application. At the same time, for those of ordinary skill in the art, according to the present application, there will be different forms of changes in the specific implementation methods and application scopes. It is not necessary and impossible to list all the implementation methods here, and the obvious changes or modifications derived therefrom are still within the scope of protection of the present application.

Claims

1. A biosensor, characterized in that: include: substrate; A power supply electrode and a plurality of working electrodes are disposed on the substrate; The substrate has a reaction area, the reaction area includes a first area and a plurality of mutually independent second areas, each of the second areas is connected to the first area, the signal end of the power electrode is located in the first area, and the signal ends of the plurality of working electrodes are respectively located in different second areas; A hydrophilic layer, wherein the hydrophilic layer is disposed on the substrate, the hydrophilic layer covers the reaction area, and one edge of the hydrophilic layer extends to the edge of the substrate; the shape of the hydrophilic layer is the same as the shape of the reaction area; A hydrophobic layer is disposed on the substrate and surrounds the hydrophilic layer; and a position between each of the second regions is filled with the hydrophobic layer.

2. The biosensor according to claim 1, characterized in that: The material of the hydrophilic layer includes: silane containing carboxyl group.

3. The biosensor according to claim 1, characterized in that: The material of the hydrophobic layer includes: silane containing fluorine or chlorine, and insulating paste.

4. The biosensor according to claim 1, characterized in that: A protective layer is provided, wherein the protective layer covers the hydrophilic layer and the hydrophobic layer, and two sides of the protective layer are connected to the substrate.

5. The biosensor according to claim 1, characterized in that: The plurality of working electrodes are located on one side or both sides of the power electrode; Alternatively, the plurality of working electrodes are evenly spaced along the circumference of the power electrode.

6. The biosensor according to claim 1, characterized in that: The power supply electrode includes: an active electrode.

7. The biosensor according to claim 1, characterized in that: The power supply electrode includes: a counter electrode and a reference electrode.

8. A method for manufacturing a biosensor, characterized in that: The method comprises: providing a substrate; A power electrode and a plurality of working electrodes are formed on the substrate by a printing process and dried, wherein the signal end of the power electrode and the signal ends of the plurality of working electrodes are located in a reaction area of ​​the substrate; the substrate has a reaction area, wherein the reaction area includes a first area and a plurality of mutually independent second areas, each of the second areas is connected to the first area, the signal end of the power electrode is located in the first area, and the signal ends of the plurality of working electrodes are respectively located in different second areas; Covering the power electrode and the leads of the plurality of working electrodes with an insulating paste to form an insulating layer, and drying the layers; forming a hydrophilic layer on the substrate, wherein the hydrophilic layer covers the reaction area, and one edge of the hydrophilic layer extends to the edge of the substrate; the shape of the hydrophilic layer is the same as that of the reaction area; forming a hydrophobic layer on the substrate, wherein the hydrophobic layer surrounds the hydrophilic layer; and the position between each of the second regions is filled with the hydrophobic layer; The reagent enzyme is printed on the plurality of working electrodes by a printing process and then dried.

9. The method for manufacturing a biosensor according to claim 8, characterized in that: Forming the hydrophilic layer on the substrate comprises: The substrate is treated with oxygen plasma to generate OH groups, thereby forming the hydrophilic layer.

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