CN112014448A - Biosensor, method for manufacturing the same, and polymer film layer for biosensor - Google Patents

Biosensor, method for manufacturing the same, and polymer film layer for biosensor Download PDF

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CN112014448A
CN112014448A CN202010906747.9A CN202010906747A CN112014448A CN 112014448 A CN112014448 A CN 112014448A CN 202010906747 A CN202010906747 A CN 202010906747A CN 112014448 A CN112014448 A CN 112014448A
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layer
polymer
biosensor
monomer
polymer film
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CN112014448B (en
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刘敏
方骏飞
许文强
黄豪
韩明松
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Shenzhen Guiji Sensing Technology Co ltd
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
    • G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage

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Abstract

The present disclosure relates to a biosensor for detecting an object in a body, including: the polymer film layer is used for controlling the diffusion of a detected object and comprises a base layer, an adhesive layer formed on the base layer and an outer surface layer which is formed on the adhesive layer and has biocompatibility, the base layer is formed by a first polymer, the adhesive layer is formed by a second polymer, the outer surface layer is formed by a third polymer, and the second polymer is a copolymer formed by a first monomer which is the same as or similar to the monomer of the first polymer and a second monomer which is the same as or similar to the monomer of the third polymer. Thus, a biosensor having a wide response linear range and having biocompatibility can be provided.

Description

Biosensor, method for manufacturing the same, and polymer film layer for biosensor
Technical Field
The present disclosure relates to a biosensor, a method of manufacturing the same, and a polymer film layer for the biosensor.
Background
Biosensors are analytical devices that combine biological, biologically derived, or biomimetic materials with optical, electrochemical, temperature, piezoelectric, magnetic, or micromechanical physicochemical sensors or sensor microsystems, and can be used to rapidly detect certain specific chemical substances in the human body, such as glucose, urea, uric acid, and a series of amino acid compounds.
In the case of an implantable in vivo current sensor, the working electrode typically includes a sensing layer in direct contact with an electrode conductive layer, when the chemical substances reach the surface of the sensing layer and are consumed, a certain linear relation exists between the detected current value and the chemical substance concentration, when the consumption capacity of the chemical substances is limited to the electrochemistry of the sensing layer, the output current value and the concentration of the chemical substances reaching the surface of the sensing layer are not in a linear relation any more, it is therefore desirable to control the concentration of the chemical species reaching the sensing layer, to extend the linear range of the response of the implanted current sensor to the chemical species, thereby enabling the implanted current sensor to detect higher glucose concentration, and because the implanted current sensor needs to be partially implanted in the body, in direct contact with the tissue, and therefore, a portion in contact with the tissue is required to have very good biocompatibility.
Disclosure of Invention
The present disclosure has been made in view of the above circumstances, and an object thereof is to provide a biosensor having a biocompatibility with an extended response linearity range, a method for manufacturing the same, and a polymer film layer for the biosensor.
To this end, a first aspect of the present disclosure provides a biosensor for detecting an object in a body, including: the biosensor comprises a substrate, an electrode arranged on the substrate, a sensing enzyme layer arranged on the electrode, and a polymer film layer arranged on the sensing enzyme layer, wherein the polymer film layer is used for controlling the diffusion of the detected object, the polymer film layer comprises a base layer, an adhesive layer formed on the base layer, and an outer surface layer which is formed on the adhesive layer and has biocompatibility, the base layer is formed by a first polymer, the adhesive layer is formed by a second polymer, the outer surface layer is formed by a third polymer, the first polymer is a homopolymer with a benzene ring or a heterocyclic ring, the second polymer is a copolymer formed by a first monomer which is the same as or similar to a monomer of the first polymer and a second monomer which is the same as or similar to a monomer of the third polymer, and the mass ratio of the first monomer to the second monomer is 3: 7 to 7: 3, in the polymer film layer, the adhesive layer has a thickness of 40 to 50% of the thickness of the polymer film layer.
In the biosensor according to the first aspect of the present disclosure, the polymeric film layer for controlling diffusion of the analyte is provided on the sensor enzyme layer of the biosensor, so that the response linearity range of the biosensor can be extended, and the outer surface layer of the polymeric film layer having biocompatibility can provide the biosensor with biocompatibility.
Further, in the biosensor according to the first aspect of the present disclosure, optionally, the first polymer is a water-swellable homopolymer, the second polymer is a water-swellable copolymer, and the third polymer is a water-soluble polymer. Therefore, the diffusion control performance and the biocompatibility of the polymer film layer can be improved, the response linear range of the biosensor can be enlarged, and the biocompatibility of the biosensor can be improved.
Further, in the biosensor according to the first aspect of the present disclosure, optionally, the water-swellable homopolymer is one selected from polystyrene, polyurethane, polyethoxyethyl acrylate, polyethoxypropyl acrylate, poly-2-vinylpyridine, poly-4-vinylpyridine, polyhydroxyethyl methacrylate, and polyhydroxyethyl acrylate, and the water-soluble polymer is one selected from polyvinylpyrrolidone, polyvinyl alcohol, chitosan, carboxymethyl chitosan, chitosan salts, alginic acid, alginate salts, hyaluronic acid, hyaluronate salts, cellulose ethers, cellulose esters, polyvinylpyrrolidone, polyacrylamide, polyacrylic acid, polyallylene alcohol, sodium polystyrene sulfonate, polyethylene glycol, and polyethylene glycol polypropylene glycol copolymer. Therefore, a polymer film layer with diffusion control performance and biocompatibility can be formed, so that the response linear range of the biosensor can be enlarged, and the biocompatibility of the biosensor can be improved.
Further, in the biosensor according to the first aspect of the present disclosure, optionally, the water-swellable copolymer is selected from the group consisting of polyethylene glycol-block-polystyrene, polyacrylic acid-co-polystyrene, polyacrylamide-block-polystyrene, polyacrylamide-co-polystyrene, poly-2-vinylpyridine-block-polystyrene, poly-4-vinylpyridine-co-polyvinylpyrrolidone, poly-2-vinylpyridine-co-polystyrene, poly-4-vinylpyridine-block-polystyrene, poly-4-vinylpyridine-co-polyacrylamide, poly-2-vinylpyridine-co-polystyrene, poly-2-vinylpyridine-co-block-polystyrene, poly-4-vinylpyridine-co-polyacrylamide, and mixtures thereof, One of polyacrylic acid ethoxy ethyl ester-copolymerization-polyhydroxyethyl acrylate and polyacrylic acid ethoxy propyl ester-copolymerization-polyvinyl alcohol. In this case, an adhesive layer having an enhanced adhesive effect with the substrate layer and the outer surface layer can be formed, which can facilitate the formation of the polymer film layer, and thus can contribute to the expansion of the response linearity range of the biosensor and the improvement of the biocompatibility of the biosensor.
Further, in the biosensor according to the first aspect of the present disclosure, optionally, the water-swellable homopolymer has a molecular weight of 50000 to 500000Da, the water-swellable copolymer has a molecular weight of 10000 to 50000Da, and the water-soluble polymer has a molecular weight of 2000 to 50000 Da. Thereby, the formation of the polymer film layer can be facilitated, thereby contributing to the expansion of the response linear range of the biosensor.
Further, in the biosensor according to the first aspect of the present disclosure, optionally, in the polymer film layer, the thickness of the substrate layer accounts for 30% to 40% of the thickness of the polymer film layer, and the thickness of the outer surface layer accounts for 20% to 30% of the thickness of the polymer film layer. In this case, the cooperation of the substrate layer, the adhesive layer and the outer skin layer can further enhance the diffusion controlling properties of the polymer film layer.
Further, in the biosensor according to the first aspect of the present disclosure, the adhesive layer is optionally adhered to the substrate layer and the outer surface layer by at least one of a conjugation effect, a similar phase dissolution, a hydrogen bonding interaction, and a cross-linking. Therefore, the bonding of the bonding layer with the substrate layer and the outer surface layer can be enhanced, so that the diffusion control performance of the polymer film layer can be improved, and the response linear range of the biosensor can be enlarged.
In the biosensor according to the first aspect of the present disclosure, the substrate layer, the adhesive layer, and the outer surface layer may be crosslinked by the same crosslinking agent, and the crosslinking agent may be at least one of an active ester, an epoxide, and a sulfate. In this case, the substrate layer, the adhesive layer, and the outer surface layer can be bonded by crosslinking, which can be advantageous in improving the diffusion control performance of the polymer film layer, and thus can be advantageous in expanding the response linearity range of the biosensor.
A second aspect of the present disclosure provides a method of manufacturing a biosensor, comprising: preparing a substrate, and arranging an electrode on the substrate; arranging a sensing enzyme layer on the electrode; and disposing a polymer film layer on the sensor enzyme layer, wherein the polymer film layer is prepared by: preparing a substrate layer agent comprising a first polymer, an adhesive layer agent comprising a second polymer and an outer surface layer agent comprising a third polymer, wherein the first polymer is a homopolymer with a benzene ring or heterocyclic ring structure, the second polymer is a copolymer formed by a first monomer which is the same as or similar to a monomer of the first polymer and a second monomer which is the same as or similar to a monomer of the third polymer, and the mass ratio of the first monomer to the second monomer is 3: 7 to 7: 3; and sequentially forming a substrate layer for controlling diffusion of chemical substances, an adhesive layer on the substrate layer and having an adhesive effect, and an outer surface layer on the adhesive layer and having biocompatibility, wherein in the polymer film layer, a thickness of the adhesive layer accounts for 40 to 50% of a thickness of the polymer film layer.
In the second aspect of the present disclosure, a polymer film layer for controlling diffusion of a test object and having biocompatibility in an outer surface layer is formed on a sensing enzyme layer of a biosensor, thereby enabling formation of a biosensor having an extended response linear range and having biocompatibility.
A third aspect of the present disclosure provides a polymer film layer for a biosensor, comprising: the adhesive layer comprises a substrate layer, an adhesive layer formed on the substrate layer and an outer surface layer which is formed on the adhesive layer and has biocompatibility, wherein the substrate layer is formed by a first polymer, the adhesive layer is formed by a second polymer, the outer surface layer is formed by a third polymer, the first polymer is a homopolymer with a benzene ring or a heterocyclic ring, the second polymer is a copolymer formed by a first monomer which is the same as or similar to a monomer of the first polymer and a second monomer which is the same as or similar to a monomer of the third polymer, the mass ratio of the first monomer to the second monomer is 3: 7-7: 3, and the thickness of the adhesive layer in the polymer film layer accounts for 40-50% of the thickness of the polymer film layer.
In the third aspect of the present disclosure, the adhesive layer is formed of the second polymer type formed by the first monomer that is the same as or similar to the monomer of the first polymer type and the second monomer that is the same as or similar to the monomer of the third polymer type, so that the firmness of the adhesive layer in adhering the substrate layer and the outer surface layer can be improved, the stability of the polymer film layer can be improved, the diffusion control performance of the polymer film layer can be improved, the concentration ratio of the test object on both sides of the polymer film layer can be fixed, and the outer surface layer with biocompatibility can improve the biocompatibility of the polymer film layer.
According to the present disclosure, a biosensor having a biocompatibility with an extended response linear range, a method of manufacturing the same, and a polymer film layer for the biosensor can be provided.
Drawings
Fig. 1 is a diagram illustrating an application scenario of a biosensor to which an example of the present disclosure relates.
Fig. 2 is a partial schematic view illustrating a probe of a biosensor according to an example of the present disclosure.
Fig. 3 is a schematic diagram illustrating the structure of a polymer film layer to which examples of the present disclosure relate.
Fig. 4 is a flowchart illustrating a method of manufacturing a biosensor according to an example of the present disclosure.
Fig. 5 is a flow chart illustrating a method of making a polymer film layer according to examples of the present disclosure.
Fig. 6 is a current curve showing the glucose biosensor measurement of example 1 of the present disclosure.
Fig. 7 is a graph showing the current versus glucose concentration in fig. 6 in a linear manner.
Fig. 8 is a section staining diagram showing the polymer film layer of example 1 of the present disclosure.
Detailed Description
Hereinafter, preferred embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. In the following description, the same components are denoted by the same reference numerals, and redundant description thereof is omitted. The drawings are schematic and the ratio of the dimensions of the components and the shapes of the components may be different from the actual ones.
Fig. 1 is a diagram showing an application scenario of a biosensor 1 according to an example of the present disclosure.
The biosensor 1 according to the present disclosure can be applied to detection of small molecular chemical substances in vivo tissues and physiological environments, for example, can be applied to detection of blood glucose (such as a glucose sensor), detection of uric acid (such as a uric acid sensor), detection of cholesterol (such as a cholesterol sensor), and the like.
In the present embodiment, the biosensor 1 may be used to detect a test object in vivo. In other words, the biosensor 1 may be for detecting an object in a body fluid. The test substance may be a chemical substance in a body fluid. For example, the test substance may be glucose, acetylcholine, amylase, bilirubin, cholesterol, chorionic gonadotropin, creatine kinase, creatine, creatinine, DNA, fructosamine, glucose, glutamine, growth hormone, ketone body, lactate, oxygen, peroxide, prostate specific antigen, prothrombin, RNA, thyroid stimulating hormone, or troponin. In addition, the test substance may be a drug in a body fluid, for example, digoxigenin, digoxin, theophylline, warfarin, or an antibiotic (such as gentamicin, vancomycin, or the like).
In the present embodiment, the biosensor 1 may include a probe P and an electronic system connected to the probe P. Wherein a part of the probe P (particularly the sensing part) can be implanted, for example, on the body surface of the human body to be in contact with the tissue fluid in the body. In addition, another part of the probe P is also connected with an electronic system positioned outside the body surface. When the biosensor 1 operates, the probe P reacts with tissue fluid or blood in the body to generate a sensing signal (e.g., a current signal), and transmits the sensing signal to an electronic system of the body surface, which processes the sensing signal to obtain the concentration of the test object. In addition, although fig. 1 shows a position where the biosensor 1 can be arranged on an arm, the present embodiment is not limited to this, and the biosensor 1 may be arranged on, for example, the abdomen, the waist, the legs, and the like.
Fig. 2 is a partial schematic view showing a probe P of the biosensor 1 according to an example of the present disclosure.
In some examples, the probe P may include a substrate 10 and an electrode 20. In other words, the biosensor 1 may include the substrate 10 and the electrode 20. Wherein the electrode 20 may be disposed on the substrate 10. In addition, the electrode 20 may serve as a sensing portion of the probe P.
In some examples, the substrate 10 may be a flexible substrate. The flexible substrate may be substantially made of at least one of Polyethylene (PE), polypropylene (PP), Polyimide (PI), Polystyrene (PS), polyethylene terephthalate (PET), polyethylene naphthalate (PEN). In addition, in other examples, the flexible substrate may also be made of substantially metal foil, ultra-thin glass, a single-layer inorganic thin film, a multi-layer organic thin film, a multi-layer inorganic thin film, or the like.
In some examples, substrate 10 may also be a non-flexible substrate. The non-flexible substrate may generally comprise a less conductive ceramic, alumina, silica, or the like. This enables implantation into a body surface (e.g., a superficial skin layer) without requiring an auxiliary implantation device (e.g., a puncture needle).
In some examples, electrode 20 may comprise a working electrode. The working electrode may be used to acquire a current signal. In addition, the electrode 20 may include a counter electrode. In other examples, the electrode 20 may also include a reference electrode, thereby enabling a three-electrode 20 sensor to be formed.
In some examples, the electrode 20 may be made of platinum, silver chloride, palladium, titanium, or iridium. This makes it possible to provide good conductivity without affecting the electrochemical reaction of the electrode 20. However, the present embodiment is not limited thereto, and in other examples, the electrode 20 may be made of at least one selected from gold, glassy carbon, graphite, silver chloride, palladium, titanium, or iridium. This can reduce the influence on the electrode 20 while having good conductivity.
In some examples, the biosensor 1 may include a sensing enzyme layer. In addition, a sensing enzyme layer may be disposed on the electrode 20 (which may be referred to as a working electrode). Further, in the biosensor 1, the electrode 20 may be wrapped or covered with a sensing enzyme layer. In some examples, the sensor enzyme layer may be formed on the electrode 20 (which may be referred to as a working electrode) by spin coating, dip-coating, drop-coating, spray-coating, or the like.
In some examples, the sensor enzyme layer may have a reactive enzyme. In some examples, the reaction enzyme in the sensor enzyme layer may be selected according to the test substance. For example, if the test substance is glucose, the reaction enzyme may be glucose oxidase or glucose dehydrogenase.
In some examples, the reaction enzyme may serve as a detection substrate for the test substance. In some examples, the reactive enzyme may chemically react with the test substance.
Hereinafter, a glucose sensor is exemplified, and GO is usedX(FAD) as glucose oxidase, illustrates the chemical reaction of glucose oxidase with glucose.
In the sensor enzyme layer, when GOX(FAD) when it encounters glucose in tissue, the following reaction occurs：
Glucose + GOx (FAD) → gluconolactone + GOx (FADH)2) … … reaction formula (I)
GOx(FADH2)+O2→GOx(FAD)+H2O2… … formula (II).
In some examples, the electrode 20 is implanted in the skin of a human body, and the reactive enzyme can continuously chemically react with the object and convert into a corresponding current signal to be transmitted to an electronic system outside the body.
In some examples, the sensor enzyme layer may have a specific cross-linking agent. This allows the reaction enzyme to be immobilized on the electrode 20 (working electrode). For example, the specific crosslinking agent may be pentanediol, bovine serum albumin-glutaraldehyde, polyepoxy compounds, 1, 4-butanediol diglycidyl ether, glutaraldehyde, reactive esters, acid anhydrides, azines, isocyanates, or acridines.
In some examples, the sensor enzyme layer may include at least one component of a metal polymer, a carbon nanotube, graphene, porous titanium dioxide, and a conductive organic salt.
In some examples, the thickness of the sensing enzyme layer may be 0.1 μm to 100 μm. In other examples, the thickness of the sensing enzyme layer may be preferably 2 μm to 10 μm. Under the condition, the thickness of the reaction enzyme is controlled within a certain degree, so that the problems that the adhesion force is reduced due to excessive reaction enzyme, the material falls off in vivo, the reaction is insufficient due to the reaction enzyme, and normal glucose concentration information cannot be fed back are solved.
In some examples, the thickness of the sensing enzyme layer may be 0.1 μm, 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 20 μm, 50 μm, 80 μm, or 100 μm.
In some examples, the biosensor 1 may include a polymer film layer 30. Specifically, in the biosensor 1, the polymer film layer 30 may be disposed on the sensor enzyme layer. In addition, the polymer film layer 30 may serve as a diffusion limiting film. In other words, the polymer film layer 30 can be used to control the diffusion of the test object.
In some examples, in the biosensor 1, the sensing enzyme layer may be wrapped or covered by the polymer film layer 30. This enables the polymer film layer 30 to control the concentration of the sample reaching the sensor enzyme layer.
In some examples, polymer film layer 30 may include a base layer 31. Wherein the base layer 31 may be disposed on the sensor enzyme layer. Additionally, in some examples, substrate layer 31 may encapsulate or cover the sensing enzyme layer.
In some examples, base layer 31 may be formed from a first type of polymer. In other examples, base layer 31 may be crosslinked from a first type of polymer. Base layer 31 may be formed from a first type of polymer that is crosslinked using a crosslinking agent.
In some examples, the first type of polymer may be a homopolymer having a benzene ring or a heterocycle. This enables formation of the base layer 31 having pores, and the object can pass through the base layer 31. In addition, the first class of polymers having a benzene ring or heterocyclic ring can facilitate the generation of a conjugation effect (e.g., pi-pi conjugation).
In some examples, base layer 31 may have diffusion-controlling properties. In other examples, substrate layer 31 may be water swellable. This improves the diffusion control performance of the underlying layer 31.
In some examples, the first type of polymer may be a water swellable homopolymer. In this case, the base layer 31 having improved diffusion control properties can be formed, so that the diffusion control properties of the polymer film layer 30 can be advantageously improved, and thus the response linearity range of the biosensor 1 can be advantageously expanded.
In some examples, the water-swellable homopolymer may be one selected from the group consisting of polystyrene, polyurethane, polyethoxyethyl acrylate, polyethoxypropyl acrylate, poly-2-vinylpyridine, poly-4-vinylpyridine, polyhydroxyethylmethacrylate, and polyhydroxyethylacrylate. This can form the base layer 31 having excellent diffusion controllability, which can contribute to improvement of the diffusion controllability of the polymer film layer 30, and can contribute to expansion of the response linearity range of the biosensor 1.
In some examples, the water-swellable homopolymer may have a molecular weight of 50000 to 500000 Da. This can facilitate the formation of the base layer 31, the formation of the polymer film layer 30, and the expansion of the response linearity range of the biosensor 1.
In some examples, the water-swellable homopolymer may have a molecular weight of 50000Da, 60000Da, 70000Da, 100000Da, 150000Da, 200000Da, 300000Da, 400000Da, or 500000 Da.
In some examples, the thickness of base layer 31 is not particularly limited. For example, base layer 31 may have a thickness of 5 μm to 20 μm. This can facilitate the incorporation of other components (e.g., the sensor enzyme layer and the adhesive layer 32).
In some examples, base layer 31 may have a thickness of 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 12 μm, 14 μm, 16 μm, 18 μm, or 20 μm.
In some examples, the polymer film layer 30 may include an adhesive layer 32. Among them, the adhesive layer 32 may be formed on the base layer 31. In addition, the adhesive layer 32 may have an adhesive function. Adhesive layer 32 may be bonded to base layer 31. In other examples, bonding layer 32 may be bonded to base layer 31 by at least one of a conjugation effect, a similar phase, hydrogen bonding interactions, and cross-linking.
In some examples, the adhesive layer 32 may be formed from a second type of polymer. The adhesive layer 32 may be formed by crosslinking the second polymer. The adhesive layer 32 may be formed by crosslinking the second polymer with a crosslinking agent.
In some examples, the second type of polymer can have a similar structure to the first type of polymer and the third type of polymer (described below), respectively. In this case, the firmness of bonding the adhesive layer 32 to the base layer 31 and the outer skin layer 33 can be improved, the stability of the polymer film layer 30 can be improved, and the base layer 31 and the outer skin layer 33 can be prevented from falling off from the adhesive layer 32, which is advantageous for improving the diffusion control performance of the polymer film layer 30.
Specifically, the second type of polymer may be a copolymer formed from a first monomer that is the same as or similar to the monomer of the first type of polymer and a second monomer that is the same as or similar to the monomer of the third type of polymer. In other words, the structure of the first monomer may be the same as or similar to the structure of the monomer of the first type of polymer, and the structure of the second monomer may be the same as or similar to the structure of the monomer of the third type of polymer.
In some examples, the mass ratio of the first monomer to the second monomer in the second polymer may be 3: 7 to 7: 3. In this case, the diffusion controlling performance of the adhesive layer 32 can be improved, and the diffusion controlling performance of the polymer film layer 30 can be advantageously improved. In other words, in the monomers forming the second type of polymer, the mass fraction of the first monomer may be 30% to 70%, and the mass fraction of the second monomer may be 30% to 70%.
In some examples, the mass ratio of the first monomer to the second monomer may be 3: 7, 7: 13, 2: 3, 9: 11, 1: 1, 11: 9, 3: 2, 13: 7, or 7: 3.
In some examples, the bonding layer 32 may have diffusion control properties. Additionally, in some examples, the adhesive layer 32 may have some water-swelling properties. This improves the diffusion control performance of the adhesive layer 32. In other examples, the second type of polymer can be a water swellable copolymer.
In some examples, the water-swellable copolymer can be selected from the group consisting of polyethylene glycol-block-polystyrene, polyacrylic acid-co-polystyrene, polyacrylamide-block-polystyrene, polyacrylamide-co-polystyrene, poly-2-vinylpyridine-block-polystyrene, poly-4-vinylpyridine-co-polyvinylpyrrolidone, poly-2-vinylpyridine-co-polystyrene, poly-4-vinylpyridine-block-polystyrene, poly-4-vinylpyridine-co-polyacrylamide, polyethyloxyethyl acrylate-co-polyhydroxyethyl acrylate, polyethyl, One of polyacrylic acid ethoxy propyl ester-copolymerization-polyvinyl alcohol. In this case, the adhesive layer 32 having an enhanced adhesive effect with the substrate layer 31 and the outer skin layer 33 can be formed, so that the formation of the polymer film layer 30 can be facilitated, and thus the range of response linearity of the biosensor 1 can be extended.
In some examples, the water-swellable copolymer may have a molecular weight of 10000 to 50000 Da. This facilitates the formation of the adhesive layer 32, the polymer film layer 30, and the expansion of the response linearity range of the biosensor 1.
In some examples, the water-swellable copolymer may have a molecular weight of 10000Da, 12000Da, 15000Da, 20000Da, 25000Da, 30000Da, 35000Da, 40000Da, or 50000 Da.
In some examples, the thickness of the adhesive layer 32 is not particularly limited. For example, the adhesive layer 32 may have a thickness of 5 μm to 20 μm. This can facilitate the fitting with other members (e.g., the base layer 31 and the outer skin layer 33).
In some examples, the adhesive layer 32 may have a thickness of 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 12 μm, 14 μm, 16 μm, 18 μm, or 20 μm.
In some examples, the polymer film layer 30 may include an outer skin layer 33. Wherein the outer skin 33 may be formed on the adhesive layer 32. In addition, the outer surface layer 33 may be biocompatible. In other examples, outer skin layer 33 may be bonded to adhesive layer 32 by at least one of a conjugation effect, a phase-similarity, a hydrogen bonding interaction, and cross-linking.
In some examples, the adhesive layer 32 may be adhered to the substrate layer 31 and the outer skin layer 33 by at least one of a conjugation effect (e.g., pi-pi conjugation), miscibility, hydrogen bonding interaction, and cross-linking. This can enhance the bonding between the adhesive layer 32 and the base layer 31 and the outer skin layer 33, which can contribute to the improvement of the diffusion control performance of the polymer film layer 30, and can contribute to the expansion of the response linearity range of the biosensor 1.
In some examples, outer skin 33 may be formed from a third type of polymer. In addition, the outer skin 33 may be crosslinked from a third type of polymer. The outer skin layer 33 may be formed by crosslinking a third polymer with a crosslinking agent.
In some examples, adhesive layer 32 and outer skin layer 33 may each be crosslinked via the same crosslinking agent. In other examples, adhesive layer 32 and substrate layer 31 may each be crosslinked via the same crosslinking agent.
In some examples, substrate layer 31, adhesive layer 32, and outer skin layer 33 may each be crosslinked via the same crosslinking agent. In this case, the substrate layer 31, the adhesive layer 32, and the outer surface layer 33 can be bonded by means of cross-linking, so that the diffusion control performance of the polymer film layer 30 can be advantageously improved, and the response linearity range of the biosensor 1 can be advantageously expanded.
In some examples, the crosslinking agent can be at least one of an active ester, an epoxide, a sulfate. For example, the crosslinking agent can be a polyisocyanate, a polyethylene glycol active ester, a glycidyl ester, a maleimide PEG active ester, polyethylene glycol ethylene oxide, 1, 4-butanediol diglycidyl ether, glutaric anhydride, 1, 4-diazide, bisacridine, or sodium sulfate.
In some examples, the third class of polymers can be biocompatible. Additionally, the third type of polymer can be a water soluble polymer. This can contribute to improving the biocompatibility of the outer surface layer 33, which can contribute to improving the biocompatibility of the polymer film layer 30 and thus the biocompatibility of the biosensor 1.
In some examples, the water-soluble polymer may be one selected from polyvinylpyrrolidone, polyvinyl alcohol, chitosan, carboxymethyl chitosan, chitosan salt, alginic acid, alginate, hyaluronic acid, hyaluronate, cellulose ethers, cellulose esters, polyvinylpyrrolidone, polyacrylamide, polyacrylic acid, polyallylamine, sodium polystyrene sulfonate, polyethylene glycol polypropylene glycol copolymer. Thereby, it is possible to facilitate the formation of the outer surface layer 33 having biocompatibility, so that the polymer film layer 30 having biocompatibility can be formed, and thus the biocompatibility of the biosensor 1 can be improved.
In some examples, the water-soluble polymer may have a molecular weight of 2000 to 50000 Da. This facilitates the film formation of the outer surface layer 33, and thus the formation of the polymer film layer 30, which in turn facilitates the improvement of the biocompatibility of the biosensor 1.
In some examples, the molecular weight of the water-soluble polymer may be 2000Da, 5000Da, 10000Da, 15000Da, 20000Da, 25000Da, 30000Da, 35000Da, 40000Da, or 50000 Da.
In some examples, the thickness of outer skin 33 is not particularly limited. For example, the thickness of the outer skin 33 may be 5 μm to 20 μm. This can facilitate the fitting with another member (for example, the adhesive layer 32). In some examples, the thickness of outer skin 33 may be 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 12 μm, 14 μm, 16 μm, 18 μm, or 20 μm.
In some examples, the thickness of the polymer film layer 30 is not particularly limited. Additionally, in some examples, the thickness of the polymer film layer 30 may be no greater than 100 μm. This can facilitate the incorporation of the polymer film layer 30 into the sensor enzyme layer. For example, the thickness of the polymer film layer 30 may be 10 μm, 15 μm, 20 μm, 30 μm, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, or 100 μm.
In some examples, the first type of polymer, the second type of polymer, and the third type of polymer may be matched to one another. For example, if the first polymer type is poly-4-vinylpyridine and the third polymer type is polyvinylpyrrolidone, the third polymer type may be poly-4-vinylpyridine-co-polyvinylpyrrolidone.
In some examples, adhesive layer 32 may wrap or cover base layer 31 in polymer film layer 30. Additionally, in some examples, in the polymer film layer 30, the outer skin layer 33 may wrap or cover the adhesive layer 32.
In the present embodiment, the diffusion controlling performance of the polymer film layer 30 can be adjusted by adjusting the thickness ratio of the substrate layer 31, the adhesive layer 32, and the outer skin layer 33. In some examples, in polymer film layer 30, the thickness of adhesive layer 32 may be no less than the thickness of substrate layer 31, and the thickness of adhesive layer 32 may be greater than the thickness of outer skin layer 33.
In some examples, in the polymer film layer 30, the adhesive layer 32 may have a thickness that is 40% to 50% of the thickness of the polymer film layer 30. This can contribute to the improvement of the diffusion control performance of the polymer film layer 30. For example, the thickness of the adhesive layer 32 may comprise 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% of the thickness of the polymer film layer 30.
In some examples, in polymer film layer 30, base layer 31 may have a thickness that is 30% to 40% of the thickness of polymer film layer 30. In this case, the combination of substrate layer 31, adhesive layer 32, and outer skin layer 33 can further enhance the diffusion controlling properties of polymer film layer 30. For example, the thickness of base layer 31 may comprise 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% of the thickness of polymer film layer 30.
In some examples, in polymer film layer 30, the thickness of outer skin layer 32 may be 20% to 30% of the thickness of polymer film layer 30. In this case, the combination of substrate layer 31, adhesive layer 32, and outer skin layer 33 can further enhance the diffusion controlling properties of polymer film layer 30. For example, the thickness of the outer skin layer 32 may account for 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% of the thickness of the polymer film layer 30.
In the present embodiment, the substrate layer 31, the adhesive layer 32, and the outer skin layer 33 may all have diffusion controlling properties. That is, the base layer 31, the adhesive layer 32, and the outer skin layer 33 can control the passage rate of the test object.
In some examples, the diffusion controlling properties of adhesive layer 32 may be matched to the diffusion controlling properties of substrate layer 31 and outer skin layer 33 in polymer film layer 30.
In some examples, the diffusion controlling properties of adhesive layer 32 may be between substrate layer 31 and outer skin layer 33. In other words, the passage rate of the test object through the adhesive layer 32 may be not less than the passage rate of the test object through the base layer 31 and not more than the passage rate of the test object through the outer skin layer 33. In this case, the adhesive layer 32 can perform a transition function to reduce the difference in diffusion control performance between the interface of the substrate layer 31 and the outer skin layer 33, thereby improving the diffusion control performance of the polymer film layer 30.
In the polymer film layer 30 according to the present embodiment, the adhesive layer 32 is formed of the second polymer type formed of the first monomer that is the same as or similar to the monomer of the first polymer type and the second monomer that is the same as or similar to the monomer of the third polymer type, so that the firmness of adhesion of the adhesive layer 32 to the substrate layer 31 and the outer surface layer 33 can be improved, the stability of the polymer film layer 30 can be improved, the diffusion control performance of the polymer film layer 30 can be advantageously improved, the concentration ratio of the test substance on both sides of the polymer film layer 30 can be fixed, and the biocompatibility of the polymer film layer 30 can be improved by the outer surface layer 33 having biocompatibility.
In the present embodiment, by providing the polymer film layer 30 for controlling the diffusion of the test object on the sensor enzyme layer of the biosensor 1, the response linearity range of the biosensor 1 can be extended, and the outer surface layer 33 having biocompatibility of the polymer film layer 30 can make the biosensor 1 biocompatible.
In the present embodiment, the upper limit of the detectable concentration of the biosensor 1 can be increased by expanding the response linearity range of the biosensor 1 using the polymer film layer 30. In some examples, the upper detectable concentration limit may be raised to 40 mM.
Hereinafter, the method of manufacturing the biosensor 1 will be described in detail with reference to the accompanying drawings. Fig. 4 is a flowchart illustrating a method of manufacturing the biosensor 1 according to an example of the present disclosure. Fig. 5 is a flow chart illustrating a method of making a polymer film layer 30 in accordance with examples of the present disclosure.
In the present embodiment, as shown in fig. 4, the method of manufacturing the biosensor 1 may include preparing the substrate 10, and disposing the electrodes 20 on the substrate 10 (step S100). In addition, the above description may be referred to with respect to the substrate 10 and the electrode 20.
In some examples, in step S100, a working electrode may be formed on the substrate 10. In addition, in step S100, a counter electrode may be formed on the substrate 10. Further, in step S100, a reference electrode may be formed on the substrate 10.
In some examples, in step S100, the electrode 20 may be formed on the substrate 10 by electroplating, evaporation, printing, extrusion, or the like.
In the present embodiment, as shown in fig. 4, the method of manufacturing the biosensor 1 may include providing a sensor enzyme layer on the electrode 20 (step S200). Specifically, in step S200, a sensor enzyme layer may be provided on the working electrode. In addition, reference may be made to the description above regarding the sensor enzyme layer.
In the present embodiment, as shown in fig. 4, the method of manufacturing the biosensor 1 may include disposing a polymer film layer 30 on the sensor enzyme layer (step S300). Reference may be made to the description above with respect to the polymer film layer 30.
In some examples, step S300 may include preparation of the polymer film layer 30 (step S310). Additionally, in some examples, as shown in fig. 5, step S310 may include preparing a base layer reagent, an adhesive layer reagent, and an outer layer reagent (step S311).
In some examples, the base layer agent may comprise a first polymer type, the adhesive layer agent may comprise a second polymer type, and the outer layer agent may comprise a third polymer type.
In some examples, the base layer reagent may comprise a cross-linking agent. In other examples, the adhesive layer reagent may include a crosslinking agent. Additionally, in some examples, the outer surface layer agent may comprise a cross-linking agent.
In some examples, in step S311, the base layer reagent may be formed by dissolving the first type of polymer in a first solvent. Specifically, in step S311, the first type polymer and the first solvent may be mixed, and subjected to ultrasonic treatment, shaking treatment, stirring treatment, and the like to obtain the basal layer reagent.
In some examples, in step S311, the first solvent may have volatility. For example, the first solvent may be an aqueous solution of ethanol, tetrahydrofuran, isopropanol, methanol, dimethylformamide, dimethylsulfoxide, or sulfolane.
In some examples, in step S311, the concentration of the first type of polymer in the base layer reagent may be 30 to 120 mg/ml. For example, the concentration of the first type of polymer may be 30mg/ml, 40mg/ml, 50mg/ml, 60mg/ml, 70mg/ml, 80mg/ml, 90mg/ml, 100mg/ml, 110mg/ml or 120 mg/ml.
In some examples, in step S311, the base layer reagent may be formed by dissolving the first type of polymer and the cross-linking agent in a first solvent. In other examples, in step S311, the mass ratio of the first polymer to the crosslinking agent may be 8: 1, 10: 1, 15: 1, 20: 1, 30: 1, 40: 1, 60: 1, 80: 1, 100: 1, 120: 1, or 128: 1.
In some examples, in step S311, the adhesive layer agent may be formed by dissolving the second type of polymer in a second solvent. Specifically, in step S311, the second polymer and the second solvent may be mixed, and subjected to ultrasonic treatment, shaking treatment, stirring treatment, or the like to obtain the adhesive layer reagent.
In some examples, in step S311, the second solvent may have volatility. For example, the second solvent may be an aqueous solution of ethanol, tetrahydrofuran, isopropanol, methanol, dimethylformamide, dimethylsulfoxide, or sulfolane. In addition, the second solvent may be the same as the first solvent.
In some examples, in step S311, the concentration of the second type of polymer in the adhesive layer reagent may be 30 to 120 mg/ml. For example, the concentration of the second type of polymer may be 30mg/ml, 40mg/ml, 50mg/ml, 60mg/ml, 70mg/ml, 80mg/ml, 90mg/ml, 100mg/ml, 110mg/ml or 120 mg/ml.
In some examples, in step S311, the base layer reagent may be formed by dissolving the second type of polymer and the cross-linking agent in a second solvent. In other examples, in step S311, the mass ratio of the second polymer to the crosslinking agent may be 8: 1, 10: 1, 15: 1, 20: 1, 30: 1, 40: 1, 60: 1, 80: 1, 100: 1, 120: 1, or 128: 1.
In some examples, in step S311, an outer skin agent may be formed by dissolving a third type of polymer in a third solvent. Specifically, in step S311, the third polymer and the third solvent may be mixed, and subjected to ultrasonic treatment, shaking treatment, stirring treatment, etc. to obtain the outer surface layer agent.
In some examples, in step S311, the third solvent may have volatility. For example, the third solvent may be an aqueous solution of ethanol, tetrahydrofuran, isopropanol, methanol, dimethylformamide, dimethylsulfoxide, or sulfolane. Additionally, in some examples, the third solvent may be the same as the first solvent. In other examples, the third solvent may be the same as the second solvent. Further, in some examples, the first solvent, the second solvent, and the third solvent may be the same.
In some examples, in step S311, the concentration of the third type of polymer in the outer skin agent may be 30 to 120 mg/ml. For example, the concentration of the third type of polymer may be 30mg/ml, 40mg/ml, 50mg/ml, 60mg/ml, 70mg/ml, 80mg/ml, 90mg/ml, 100mg/ml, 110mg/ml or 120 mg/ml.
In some examples, in step S311, an outer skin agent may be formed by dissolving a third type of polymer and a crosslinker in a third solvent. In other examples, in step S311, the mass ratio of the third type polymer to the crosslinking agent may be 8: 1, 10: 1, 15: 1, 20: 1, 30: 1, 40: 1, 60: 1, 80: 1, 100: 1, 120: 1, or 128: 1.
In some examples, as shown in fig. 5, step S310 may include sequentially forming a substrate layer 31, an adhesive layer 32, and an outer skin layer 33 (step S312).
In some examples, in step S312, the substrate layer 31, the adhesive layer 32, and the outer surface layer 33 may be sequentially formed by spin coating, dip drawing, drop coating, spray coating, etc. to form the polymer film layer 30. In addition, the polymer film layer 30 may be dried in a nitrogen atmosphere.
In some examples, in step S300, the electrode 20 (which may be referred to as a working electrode) may be sequentially subjected to a pull-up dipping process through a base layer reagent, an adhesive layer reagent, and an outer surface layer reagent to form the polymer film layer 30 on the electrode 20.
In the present embodiment, the polymer film layer 30 for controlling the diffusion of the test object and having biocompatibility in the outer surface layer 33 is formed on the sensor enzyme layer of the biosensor 1, thereby enabling the formation of the biosensor 1 having an extended response linear range and biocompatibility.
According to the present disclosure, it is possible to provide a biosensor 1 having a biocompatibility with an extended response linear range, a method of manufacturing the same, and a polymer film layer 30 for the biosensor 1.
Hereinafter, embodiments of the present invention will be described in further detail with reference to specific examples. Fig. 6 is a current curve showing the glucose biosensor measurement of example 1 of the present disclosure. Fig. 7 is a graph showing the current versus glucose concentration in fig. 6 in a linear manner. Fig. 8 is a section staining diagram showing the polymer film layer of example 1 of the present disclosure.
[ examples ] A method for producing a compound
In the present embodiment, a glucose biosensor of a three-electrode system with glucose oxidase is used as the biosensor. In the raw materials, the molecular weight of poly-4-vinylpyridine is 50000Da, the molecular weight of polyvinylpyrrolidone is 5000Da, the molecular weight of poly-4-vinylpyridine-co-polyvinylpyrrolidone is 10000Da, the molecular weight of polyethoxyethyl acrylate is 15000Da, the molecular weight of polyethoxyethyl acrylate-co-polyhydroxyethyl acrylate is 10000Da, the molecular weight of polyacrylamide is 8000Da, and the molecular weight of poly-4-vinylpyridine-co-polyacrylamide is 12000 Da.
Firstly, preparing a base layer reagent raw material of each example (example 1 to example 3) according to table 1, dissolving a first polymer in a first solvent, and performing ultrasonic treatment and oscillation until the first polymer is completely dissolved to obtain a base layer reagent; next, adhesive layer reagent raw materials of the respective examples (examples 1 to 3) were prepared according to table 1, and the second polymer was dissolved in the second solvent, and ultrasonically vibrated until completely dissolved to obtain an adhesive layer reagent; then, the raw materials of the outer surface layer reagents of the respective examples (examples 1 to 3) were prepared according to table 1, and the third type of polymer was dissolved in the third solvent, and then sonicated and shaken until completely dissolved, thereby obtaining the outer surface layer reagents.
Then, the working electrode of the glucose biosensor of each example was sequentially dipped in the basal layer reagent, the adhesive layer reagent, and the outer layer reagent by pulling for 5 seconds at intervals of 10 minutes, and each reagent was dipped 12 times to prepare a polymer film 30, which was then dried in a nitrogen atmosphere for 48 hours to complete the preparation.
Finally, the following two tests were performed: testing the performance test of the glucose biosensor obtained in each example; the firmness test for bonding the substrate layer, the bonding layer and the outer surface layer in the polymer film layers in each example was carried out by the following specific steps:
(1) the glucose biosensor was immersed in standard PBS buffer (pH 7.4, 150mM NaCl) followed by an initial pulse of 360s of 1.1 volts. Each glucose biosensor was subjected to the remaining measurement at 0.05V, waiting 30 minutes for the glucose biosensor to reach a constant background, and glucose was added to the solution at concentrations of 0mM, 5mM, 10mM, 15mM, 20mM, 25mM, 30mM, 35mM, 40mM to measure the linearity of the reaction, the solution was allowed to equilibrate for 3 minutes after each addition of glucose, and the solution was continuously stirred during the measurement to make its concentration uniform. The glucose biosensor of each example showed good linearity of the response to glucose, and specifically, as can be seen from fig. 6 and 7, the glucose sensor of example 1 showed good linear correlation of the glucose concentration and the response current in the range of 0mM to 40 mM.
(2) Fixing a working electrode of the glucose biosensor in an epoxy resin solution by using a clamp, curing and molding, and grinding and cutting in a grinding machine after 24 hours, wherein the grinding and cutting depth is one half of the total width of the working electrode; then washing the mixture by using deionized water, and then baking the mixture in a 45-degree oven for 60 minutes; then immersing the dried working electrode into 0.1% nile red ethanol solution for 5s, washing with ethanol and drying at room temperature; finally, the section is observed under an optical microscope with a magnification of 500. The base layer, the adhesive layer and the outer surface layer of the polymer film layer in each example are firmly bonded, specifically, as can be seen from fig. 8, the dye adsorption degree of each layer in the polymer film layer in example 1 is different, the layers can be clearly distinguished from each other in terms of color, and after being ground and cut by a grinding machine, no obvious crack exists among the base layer, the adhesive layer and the outer surface layer, and the bonding among the layers is firmly bonded.
TABLE 1
Figure BDA0002661744980000181
Figure BDA0002661744980000191
While the present disclosure has been described in detail in connection with the drawings and examples, it should be understood that the above description is not intended to limit the disclosure in any way. Those skilled in the art can make modifications and variations to the present disclosure as needed without departing from the true spirit and scope of the disclosure, which fall within the scope of the disclosure.

Claims (10)

1. A biosensor for detecting a subject in a body, comprising: the biosensor comprises a substrate, an electrode arranged on the substrate, a sensing enzyme layer arranged on the electrode, and a polymer film layer arranged on the sensing enzyme layer, wherein the polymer film layer is used for controlling the diffusion of the detected object, the polymer film layer comprises a base layer, an adhesive layer formed on the base layer, and an outer surface layer which is formed on the adhesive layer and has biocompatibility, the base layer is formed by a first polymer, the adhesive layer is formed by a second polymer, the outer surface layer is formed by a third polymer, the first polymer is a homopolymer with a benzene ring or a heterocyclic ring, the second polymer is a copolymer formed by a first monomer which is the same as or similar to a monomer of the first polymer and a second monomer which is the same as or similar to a monomer of the third polymer, and the mass ratio of the first monomer to the second monomer is 3: 7 to 7: 3, in the polymer film layer, the adhesive layer has a thickness of 40 to 50% of the thickness of the polymer film layer.
2. The biosensor of claim 1,
the first polymer is a water-swellable homopolymer, the second polymer is a water-swellable copolymer, and the third polymer is a water-soluble polymer.
3. The biosensor of claim 2,
the water swelling homopolymer is one selected from polystyrene, polyurethane, polyethoxyethyl acrylate, polyethoxypropyl acrylate, poly-2-vinylpyridine, poly-4-vinylpyridine, polyhydroxyethyl methacrylate and polyhydroxyethyl acrylate, and the water soluble polymer is one selected from polyvinylpyrrolidone, polyvinyl alcohol, chitosan, carboxymethyl chitosan, chitosan salt, alginic acid, alginate, hyaluronic acid, hyaluronate, cellulose ethers, cellulose esters, polyacrylamide, polyacrylic acid, polypropylene alcohol, polystyrene sodium sulfonate, polyethylene glycol and polyethylene glycol polypropylene glycol copolymer.
4. The biosensor according to claim 2 or 3,
the water swelling copolymer is selected from polyethylene glycol-block-polystyrene, polyacrylic acid-copolymerization-polystyrene, polyacrylamide-block-polystyrene, polyacrylamide-copolymerization-polystyrene, poly 2-vinylpyridine-block-polystyrene, poly 4-vinylpyridine-copolymerization-polyvinylpyrrolidone, poly 2-vinylpyridine-copolymerization-polystyrene, poly 4-vinylpyridine-block-polystyrene, poly 4-vinylpyridine-copolymerization-polyacrylamide, polyethoxyethyl acrylate-copolymerization-polyhydroxyethyl acrylate, polyethylene glycol-co-polystyrene, polyethylene glycol-co-, One of polyacrylic acid ethoxy propyl ester-copolymerization-polyvinyl alcohol.
5. The biosensor of claim 2,
the molecular weight of the water swelling homopolymer is 50000-500000 Da, the molecular weight of the water swelling copolymer is 10000-50000 Da, and the molecular weight of the water soluble polymer is 2000-50000 Da.
6. The biosensor of claim 1,
in the polymer film layer, the thickness of the substrate layer accounts for 30% to 40% of the thickness of the polymer film layer, and the thickness of the outer skin layer accounts for 20% to 30% of the thickness of the polymer film layer.
7. The biosensor of claim 1,
the adhesive layer is adhered to the substrate layer and the outer skin layer by at least one of a conjugation effect, a similar phase dissolution, a hydrogen bonding interaction, and a cross-linking.
8. The biosensor of claim 1,
the substrate layer, the bonding layer and the outer surface layer are respectively formed by cross-linking the same cross-linking agent, and the cross-linking agent is at least one of active esters, epoxides and sulfates.
9. A method for manufacturing a biosensor is provided,
the method comprises the following steps:
preparing a substrate, and arranging an electrode on the substrate;
arranging a sensing enzyme layer on the electrode; and is
A polymer film layer is arranged on the sensing enzyme layer,
wherein the polymer film layer is prepared by the following steps:
preparing a substrate layer agent comprising a first polymer, an adhesive layer agent comprising a second polymer and an outer surface layer agent comprising a third polymer, wherein the first polymer is a homopolymer with a benzene ring or heterocyclic ring structure, the second polymer is a copolymer formed by a first monomer which is the same as or similar to a monomer of the first polymer and a second monomer which is the same as or similar to a monomer of the third polymer, and the mass ratio of the first monomer to the second monomer is 3: 7 to 7: 3; and is
Sequentially forming a substrate layer, an adhesive layer on the substrate layer and having an adhesive effect, and an outer surface layer on the adhesive layer and having biocompatibility, wherein in the polymer film layer, the thickness of the adhesive layer accounts for 40% to 50% of the thickness of the polymer film layer.
10. A polymer film layer for a biosensor, comprising,
the method comprises the following steps: the adhesive layer comprises a substrate layer, an adhesive layer formed on the substrate layer and an outer surface layer which is formed on the adhesive layer and has biocompatibility, wherein the substrate layer is formed by a first polymer, the adhesive layer is formed by a second polymer, the outer surface layer is formed by a third polymer, the first polymer is a homopolymer with a benzene ring or a heterocyclic ring, the second polymer is a copolymer formed by a first monomer which is the same as or similar to a monomer of the first polymer and a second monomer which is the same as or similar to a monomer of the third polymer, the mass ratio of the first monomer to the second monomer is 3: 7-7: 3, and the thickness of the adhesive layer in the polymer film layer accounts for 40-50% of the thickness of the polymer film layer.
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CN116359311A (en) * 2023-02-15 2023-06-30 重庆联芯致康生物科技有限公司 Dynamic lactic acid sensor film and preparation method thereof
CN116359311B (en) * 2023-02-15 2024-05-28 重庆联芯致康生物科技有限公司 Dynamic lactic acid sensor film and preparation method thereof
CN116735885A (en) * 2023-05-25 2023-09-12 深圳硅基传感科技有限公司 Multi-analyte monitoring sensor
CN117686564A (en) * 2024-02-01 2024-03-12 深圳硅基传感科技有限公司 Biosensor and isolation layer and analyte monitoring device thereof

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