Flux-restricted polymer film

Abstract

The present invention relates generally to flux-limiting polymer membranes for analyte sensors and analyte sensors including flux-limiting polymer membranes.

Description

[Technical Field]

[0001] The present invention generally relates to flux-restricted polymer membranes for sample sensors, and sample sensors comprising flux-restricted polymer membranes. [Background technology]

[0002] Monitoring specific bodily functions, more specifically monitoring the concentration of one or more specific samples, plays a crucial role in the prevention and treatment of various diseases.

[0003] Along with so-called point measurements, which involve specifically collecting bodily fluid samples from users and investigating sample concentrations, continuous measurements are becoming increasingly available. Therefore, there is a growing demand for accurate sample sensors that enable reliable and cost-effective sample detection from bodily fluids or other samples. Sample sensors for determining sample concentrations under in vivo conditions are known from International Publication 2010 / 028708A1. Another example of such a sensor is disclosed in International Publication 2012 / 130841A1. Furthermore, International Publication 2007 / 147475A1 discloses an amperometric sensor implanted in the body to measure the concentration of a sample in bodily fluids. Alternative sensor elements are disclosed in International Publication 2014 / 001382A1.

[0004] International Publication No. 03 / 085372 relates to biosensor membranes composed of heterocyclic nitrogen-containing polymers. While these polymers have been found useful as flux (or diffusion) limiting membranes for sample sensors, they often exhibit undesirable long run-in times and / or significant changes in sensitivity throughout their lifetime. This is problematic, especially when sensors are stored for extended periods before use.

[0005] The problem that the present invention aims to solve is to provide a sample sensor comprising a working electrode and a flux-restricting polymer film disposed on the working electrode, which avoids the aforementioned drawbacks. In particular, the present invention aims to provide a sample sensor that avoids its lifespan.Improved stability over time and / or short run-in time has The objective is to provide sensors.

[0006] Therefore, it is desirable to provide a sensor that addresses the aforementioned technical challenges. [Overview of the project]

[0007] The above-mentioned problems are addressed by a sample sensor comprising a working electrode and a flux-restricting polymer film disposed on the working electrode, having the features of an independent claim. Advantageous embodiments that can be realized individually or in any combination are enumerated in the dependent claims and throughout this specification.

[0008] The sample sensor according to the present invention maintains sufficient sensitivity while having a long lifespan. Improved stability over time and / or during short run-ins interval Having it is advantageous.

[0009] According to a first aspect of the present invention, a sample sensor is provided comprising at least one working electrode and a flux-restricting polymer membrane disposed on the at least one working electrode, wherein the flux-restricting polymer membrane is given by the following formula (I): TIFF0007871259000001.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. It contains a poly(vinylpyridine) polymer having the following properties.

[0010] A further aspect of the present invention relates to a polymer membrane for use in a sample sensor, and more particularly for use as a flux-limiting polymer membrane in a sample sensor, the polymer membrane comprising a polymer having the above formula (I).

[0011] Further aspects of the present invention include a polymer having the above formula (I) and Crosslinking agent and Solvent and, This relates to a liquid composition containing [a specific compound / substance].

[0012] Yet another aspect of the present invention is a method for manufacturing a sample sensor, a) A step of providing a substrate having a first side surface and a second side surface, and at least one working electrode disposed on the first side surface of the substrate, b) A step of forming a layer of flux-restricting polymer film on at least one working electrode, wherein the flux-restricting polymer film comprises a polymer having the above formula (I), Regarding methods including

[0013] In a particular embodiment, x is approximately 5 mol%, y is approximately 85 mol%, and z is approximately 10 mol%.

[0014] definition As used below, the terms “have,” “comprise,” or “include,” or any grammatical variations thereof, are used non-exclusively. Therefore, these terms may refer to both situations in which the entity described in this context has no further features beyond those introduced by these terms, and situations in which one or more additional features exist. For example, the expressions “A has B,” “A comprises B,” and “A includes B” may refer to both situations in which A has no other elements besides B (i.e., A consists solely of B and exclusively of B), and situations in which entity A has one or more additional elements besides B, such as element C, elements C and D, and further elements.

[0015] Furthermore, it should be noted that terms such as "at least one", "one or more" or similar expressions indicating that a feature or element may be present one or more times are typically only used once when introducing each feature or element. In the following, in most cases, when referring to each feature or element, the expressions "at least one" or "one or more" are not repeated, despite the fact that each feature or element may be present one or more times.

[0016] Furthermore, when used hereinafter, terms such as "preferably", "more preferably", "particularly", "more particularly", "specifically", "more specifically" or similar terms are used in conjunction with any feature without limiting the possibility of alternatives. Accordingly, the features introduced by these terms are any features and are not intended to limit the claims in any way. As will be appreciated by those skilled in the art, the present invention may be implemented by using alternative features. Similarly, features introduced by "in an embodiment of the present invention" or similar expressions are any features without any limitation regarding alternative embodiments of the present invention, without any limitation regarding the scope of the present invention, and without any limitation regarding the possibility of combining such features introduced in such a way with any other or non - arbitrary features of the present invention.

Embodiments for Carrying out the Invention

[0017] The present invention relates to a specimen sensor comprising at least one working electrode and a flux - limiting polymer film disposed on the at least one working electrode, a polymer film for use in a specimen sensor, a liquid composition comprising a polymer, a cross - linking agent and a solvent, and a method for manufacturing a specimen sensor.

[0018] As used herein, the term "analyte sensor" is a broad term and should be given its ordinary and customary meaning to those of ordinary skill in the art and should not be limited to a special or customized meaning. Specifically, but not limited to, this term can refer to any element or device configured to detect or measure the concentration of at least one analyte. The analyte sensor can specifically be an analyte sensor suitable for at least partial implantation into a user's body tissue, more specifically, an analyte sensor for continuous monitoring of an analyte.

[0019] In certain embodiments, the analyte sensor of the present invention is an electrochemical sensor that includes a working electrode, at least one additional electrode, and respective circuitry. More specifically, the sensor is an amperometric electrochemical sensor that includes at least one working electrode. Typically, the analyte sensor includes at least one additional electrode, particularly a counter electrode and / or a reference electrode or a counter / reference composite electrode.

[0020] In certain embodiments, the analyte sensor is a two-electrode sensor that particularly includes one working electrode and one counter / reference composite electrode.

[0021] As used herein, the term “working electrode” is a broad term and should be given its usual and customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. Specifically, the term may refer to an electrode of a sample sensor that is sensitive to a sample, but is not limited to this. The working electrode may be located on at least one first side surface of at least one substrate. In particular, the working electrode comprises at least one conductive material and at least one sensing material, the at least one sensing material being coated on a coated area on the conductive material on the first side surface of the sensor substrate. The working electrode is sensitive to a sample that can be measured by a polarization voltage applied between the working electrode and a reference electrode and which can be adjusted by a potentiostat. The measurement signal may be provided as a current between the counter electrode and the working electrode. A separate counter electrode may not be present, and a pseudo-reference electrode may be present which can also function as a counter electrode. Thus, a sample sensor may typically comprise at least two sets of electrodes, and in one embodiment, three sets of electrodes. In particular, the sensing material is present only on the working electrode.

[0022] The layer of sensing material may be present only on the working electrode and typically not on any further electrodes; for example, the counter electrode and / or reference electrode may not contain a layer of sensing material.

[0023] In particular, the specimen sensor according to the present invention may be fully or partially implanted and thus adapted to perform detection of specimens in body fluids, particularly interstitial fluid, within subcutaneous tissue. Other parts or components may remain outside the body tissue. For example, as used herein, the terms “implantable” or “subcutaneous” mean being fully or at least partially positioned within the user’s body tissue. For this purpose, the specimen sensor may comprise an insertable portion, and the term “insertable portion” may generally refer to a part or component of an element configured to be insertable into any body tissue. The insertable portion may include a working electrode and typically at least one further electrode, e.g., a counter electrode, a reference electrode and / or a pair / reference composite electrode. In certain embodiments, the working electrode is positioned on a first side of the substrate, at least one further electrode is positioned on a second side of the substrate, and all electrodes are positioned in the insertable portion. The uninserted portion of the sensor is the top of the sensor and may include contacts for connecting the sensor to an electronic unit.

[0024] Furthermore, as used herein, the term “specimen” is a broad term and should be given its usual customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. Specifically, the term may refer to any element, component, or compound that may be present in body fluids and whose concentration may be of interest to the user, but is not limited to this. Specifically, a specimen may be, or may contain, any chemical substance or chemical compound that may be involved in the user’s metabolism, such as at least one metabolite. As an example, at least one metabolite may be selected from the group consisting of glucose, cholesterol, triglycerides, and lactate, and more specifically, the specimen may be glucose. However, additionally or alternatively, other types of specimens and / or any combination of specimens may be determined.

[0025] The sample sensor may be configured for at least partial implantation into the user's body tissue, specifically percutaneous insertion; more specifically, the sample sensor may be configured for continuous monitoring of a sample; and even more specifically, the sample sensor may be configured for continuous glucose monitoring.

[0026] The sample sensor of the present invention comprises at least one flux-restricted polymer membrane, the at least one flux-restricted polymer membrane positioned on the working electrode, meaning that the at least one flux-restricted polymer membrane at least partially covers the working electrode. In one embodiment of the present invention, the flux-restricted membrane completely covers the working electrode. The flux-restricted polymer membrane can generally allow one or more molecules and / or compounds to pass through selectively, while other molecules and / or compounds are stopped by the membrane. Specifically, the flux-restricted polymer membrane is permeable to at least one sample to be detected. For example, the membrane can be permeable to one or more of glucose, lactate, cholesterol, or other types of samples. Thus, the at least one flux-restricted polymer membrane can function as a diffusion barrier controlling the diffusion of the sample from the outside, for example, the bodily fluids surrounding the sample sensor, to the detection material, i.e., enzyme molecules in the detection material.

[0027] In certain embodiments, the flux-restricted polymer membrane is glucose-permeable. For example, the flux-restricted polymer membrane is at least about 1 × 10⁻⁶ -6 cm 2 / s, preferably about 1 × 10 -6 cm 2 / s~approx. 1×10 -10 cm 2 It may have a glucose diffusion coefficient of / s. The glucose diffusion coefficient can be determined by diffusion cells and flux-limiting film foils manufactured by the doctor blade method on polypropylene.

[0028] The flux-restricted polymer film of the present invention is represented by the following formula (I): TIFF0007871259000002.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. It contains a poly(vinylpyridine) polymer having the following properties.

[0029] The polymer of formula (I) comprises two or three different monomer units, a 4-sulfonatopropyl-vinylpyridinium unit, a 4-vinylpyridine unit, and optionally a styrene unit. In certain embodiments, the polymer consists of the two or three different monomer units described above. The parameters x, y, and z in formula (I) represent the relative molar amount (i.e., mol%) of each monomer unit in the polymer.

[0030] In the present invention, parameter x corresponds to the relative molar amount of 4-sulfonatopropyl-vinylpyridinium units and is in the range of about 2 to about 8 mol%, about 3 to about 7 mol%, about 4 to about 6 mol%, and particularly about 5 mol%.

[0031] Parameter y corresponds to the relative molar amount of 4-vinylpyridine units and ranges from approximately 72 to approximately 98 mol%, 82 to approximately 88 mol%, approximately 83 to approximately 87 mol%, and approximately 84 to approximately 86 mol%, particularly approximately 85 mol%.

[0032] The parameter z corresponds to the relative amount of styrene units and ranges from 0 to about 20 mol%, about 7 to about 13 mol%, about 8 to about 12 mol%, about 9 to about 11 mol%, and especially about 10 mol%.

[0033] The sum of the mole percentages of parameters x, y, and z is usually 100 mole percent.

[0034] In certain embodiments, the polymer of formula (I) is a statistical copolymer, i.e., individual monomer units are randomly incorporated into the polymer chain.

[0035] In certain embodiments, the polymer has a weight-average molecular weight of about 60 kDa to about 200 kDa, about 80 kDa to about 160 kDa, about 100 kDa to about 140 kDa, and particularly about 120 kDa. In certain embodiments, the polymer has a number-average molecular weight of about 40 kDa to about 90 kDa, about 50 kDa to about 70 kDa, and particularly about 60 kDa. In certain embodiments, the polymer has a polydispersity index of about 1.4 to about 3, and particularly about 2.

[0036] In certain embodiments, the polymer has the weight-average molecular weight, number-average molecular weight, and optionally a polydispersity index.

[0037] The molecular weights mentioned above pertain to the uncrosslinked polymer of formula (I). Weight-average and number-average molecular weights are measured by GPC using PMMA (polymethyl methacrylate) as a standard. Polydispersity is the ratio of weight-average molecular weight to number-average molecular weight.

[0038] The polymer of formula (I) can be produced by known methods, for example, as described in International Publication No. 03 / 085372, by reacting a poly(4-vinylpyridine-co-styrene) copolymer, particularly a statistical poly(4-vinylpyridine-co-styrene) copolymer, containing, for example, a relative amount of x+y mol% of 4-vinylpyridine units and a relative amount of z mol% of styrene units (wherein x, y, and z are defined above), with 1,3-propanesultone under conditions that 4-sulfonatopropyl-vinylpyridinium units are formed in a relative amount of x mol%.

[0039] In certain embodiments, the polymer of formula (I) in a flux-restricted polymer film is crosslinked by a crosslinking agent. The crosslinking agent is a molecule containing at least two functional groups that can link at least two molecules together or at least two parts of the same molecule together. Linking at least two molecules is called intermolecular crosslinking, and linking at least two parts of the same molecule is called intramolecular crosslinking. A crosslinking agent having three or more functional groups may perform both intermolecular and intramolecular crosslinking simultaneously.

[0040] The crosslinking agent can react with functional groups on the polymer, particularly alkyl-sulfonate groups and / or pyridine groups. The polymer in the flux-restricted polymer film of the present invention is crosslinked with a certain amount of crosslinking agent to provide sufficient crosslinking of the polymer. For example, the weight ratio of polymer to crosslinking agent in the flux-restricted polymer film is about 8:1 (w / w) to about 16:1 (w / w), about 10:1 (w / w) to about 14:1 (w / w), and particularly about 12:1 (w / w).

[0041] Specifically, the flux-restricted polymer film may contain at least one crosslinking agent selected from UV-curable crosslinking agents and chemical crosslinking agents. More specifically, the detection material contains a chemical crosslinking agent.

[0042] As used herein, the term “chemical crosslinking agent” is a broad term and should be given its usual customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. Specifically, the term may refer to a crosslinking agent that, when exposed to heat, can initiate a chemical reaction that produces a crosslinked molecular network and / or crosslinked polymer. “Exposed to heat” may refer to exposure to a temperature above 15°C, specifically above 20°C, more specifically to a temperature in the range of 20°C to 50°C, and even more specifically to a temperature in the range of 20°C to 25°C. More specifically, a chemical crosslinking agent may initiate crosslinking of layers of the sensing material when exposed to heat.

[0043] Suitable chemical crosslinking agents according to the present invention include epoxide-based crosslinking agents, such as diglycidyl ethers including poly(ethylene glycol) diglycidyl ether (PEG-DGE) and poly(propylene glycol) diglycidyl ether; trifunctional short-chain epoxides; anhydrides; resorcinol diglycidyl ether; bisphenols, such as bisphenol A diglycidyl ether, diglycidyl 1,2-cyclohexane dicarboxylate, poly(ethylene glycol) diglycidyl ether, glycerol diglycidyl ether, 1,4-butanediol diglycidyl ether, and poly(propylene glycol) diglycidyl Diglycidyl ethers such as diglycidyl ethers, poly(dimethylsiloxane), diglycidyl ether, neopentyl glycol diglycidyl ether, 1,2,7,8-diepoxyoctane, and 1,3-glycidoxypropyl-1,1,3,3-tetramethyldisiloxane; triglycidyl ethers such as N,N-diglycidyl-4-glycidyloxyaniline and trimethylolpropane triglycidyl ether; and tetraglycidyl ethers, selected from, for example, tetrakis epoxycyclosiloxane, pentaerythritol tetraglycidyl ether, and tetraglycidyl-4,4'-methylenebisbenzeneamine.

[0044] In certain embodiments, the chemical crosslinking agent is PEG-DGE having a number average molecular weight of about 200 Da or more, for example, PEG-DGE having a number average molecular weight of about 200 Da. In further embodiments, the crosslinking agent is N,N-diglycidyl-4-glycidyloxyaniline. For example, the crosslinking agent may be selected from the group consisting of PEG-DGE and N,N-diglycidyl-4-glycidyloxyaniline having a number average molecular weight of 200 Da.

[0045] As used herein, the term “UV-curable” is a broad term and should be given its usual and customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. Specifically, the term may refer to the ability of a chemical substance, e.g., a crosslinking agent, to initiate a photochemical reaction that generates a crosslinked molecular network and / or crosslinked polymer when irradiated with light in the UV spectral range, but not limited to this. More specifically, a UV-curable crosslinking agent may initiate crosslinking of layers of a sensing material when irradiated with UV light. Crosslinking may be initiated in particular as described below herein.

[0046] Suitable UV-curable crosslinking agents according to the present invention include benzophenone, diaziline, and azide. Particularly suitable UV-curable crosslinking agents are selected from the group consisting of reaction products of the reaction of benzophenone containing a crosslinking agent, poly(di(2-hydroxy3-aminobenzophenone propylene) glycol), dibenzophenone 1,2-cyclohexane dicarboxylate, bis[2-(4-azidosalicylamido)ethyl] disulfide, 4-aminobenzophenone with any one of the diglycidyl crosslinking agents, triglycidyl crosslinking agents, and tetraglycidyl crosslinking agents mentioned above. An example of such a reaction product is the reaction product of 2,4,6,8-tetramethyl-2,4,6,8-tetrakis(2-hydroxy3-aminopropylbenzophenone)-cyclotetrasiloxane, 4-benzoylbenzoate N-succinimidyl ester with a diamine or jefamine.

[0047] In certain embodiments, the crosslinking agent comprises two, three or more functional groups. A specific example of a functional group is an epoxide group. Preferably, the crosslinking agent comprises at least two functional epoxide groups, more preferably three functional epoxide groups. More preferably, the crosslinking agent is a triglycidyl ether. A particularly preferred example of the crosslinking agent is N,N-diglycidyl-4-glycidyloxyaniline.

[0048] The flux limiting polymer membrane can have a thickness sufficient to provide mechanical stability. Specifically, the flux limiting polymer membrane can have a thickness of from about 1 μm to about 150 μm.

[0049] In addition to the polymers and crosslinkers described above, the flux limiting polymer membrane may contain further components, for example polymer components and / or non-polymer components, and the non-polymer components may be dispersed and / or dissolved in the polymer. Examples of non-polymer components include plasticizers, particularly biocompatible plasticizers such as tri-(2-ethylhexyl) trimellitate and / or glycerol.

[0050] Further components optionally present in the polymer membrane are salts containing pharmaceuticals, corticosteroids, heparin and ions such as Na + , Cl - and / or Br - and the like.

[0051] In certain embodiments, the membrane comprises the poly(vinylpyridine)-based polymer of formula (I) as the sole polymer component.

[0052] The flux limiting polymer membrane of the present invention may be applied to the analyte sensor as a liquid composition by a coating process, particularly a wet coating process.

[0053] As used herein, the term “coating process” may refer to any process for applying at least one layer to at least one surface of any object. The applied layer may completely cover the object, for example, a working electrode, or it may cover only a portion of the object. The layer may be applied by a coating process in which the material is provided, for example, in liquid form, exemplary as a suspension or solution, and can be distributed onto the surface. Specifically, a coating process may include a wet coating process selected from the group consisting of: spin coating; spray coating; doctor blade method; printing; distribution; slot coating; and dipping coating. Preferred wet coating processes are dipping coating or spray coating.

[0054] After application, the liquid composition may be subjected to at least one curing step, in which at least a portion of the polymer is crosslinked. The terms “crosslinking” and “curing” are used interchangeably herein. A suitable method for initiating crosslinking depends on the type of crosslinking agent and is known to those skilled in the art. Since preferred crosslinking agents are chemical crosslinking agents, curing is preferably carried out without UV light, at essentially ambient temperature or up to about 90°C. As used herein, the term “ambient temperature” is understood specifically as a temperature of 15°C to 30°C, more specifically 20°C to 25°C. Curing with UV-curable crosslinking agents is generally induced by irradiation with UV light. As used herein, the term “UV light” generally refers to electromagnetic radiation in the ultraviolet spectral range. The term “ultraviolet spectral range” usually refers to electromagnetic radiation in the range of 1 nm to 380 nm, preferably light in the range of 100 nm to 380 nm. Curing can usually be carried out at room temperature.

[0055] Furthermore, after application, the liquid composition may be subjected to at least one drying step. Specifically, especially when using a chemical crosslinking agent, the curing step and drying may be performed simultaneously. Alternatively, the drying step may be performed after the curing step. Specifically, the composition may be dried at ambient temperature or up to about 50°C for about 10 minutes or less, or about 5 minutes or less, for example, about 0.5 to about 10 minutes.

[0056] The sample sensor of the present invention includes at least one working electrode. In a particular embodiment, the sample sensor is (i) A substrate, The first aspect and the second aspect, A substrate comprising at least one conductive material disposed on a first side surface of the substrate, (ii) A working electrode comprising a sensing material that at least partially covers a first side surface of a substrate, wherein the sensing material comprises at least one enzyme, (iii) A polymer film comprising a polyvinylpyridine polymer of formula (I), wherein the film at least partially covers the working electrode, and Includes.

[0057] As used herein, the term “substrate” is a broad term and should be given its usual and customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. The term “substrate” is used synonymously with the term “sensor substrate” and may specifically refer to any type of material or combination of materials suitable for forming a polymer layer covering the working electrodes described herein, but not limited to such materials. In particular, as understood herein, “sensor substrate” may include electrical insulating materials.

[0058] As used herein, the term “layer” is a broad term and should be given its usual customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. The term may, in particular, but not limited to, elements of the layer configuration of a sample sensor. Specifically, the term “layer” may refer to any substrate, specifically any coating of a flat substrate. Specifically, a layer may have an outer extension exceeding at least twice, at least five times, at least ten times, and even at least twenty times or more in thickness. Specifically, a sample sensor may have a layer configuration. A sample sensor may include multiple layers, such as at least one conductive material, at least one layer of at least one sensing material, and optionally at least one film layer. One or more layers of a sample sensor may include sublayers. For example, a layer containing a conductive material may include at least one further layer.

[0059] As used herein, the term “electrical insulating material” is a broad term and should be given its usual customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. “Electrical insulating material” may also refer to dielectric materials. Specifically, the term may refer to materials or combinations of materials that prevent the transfer of electric charge and do not sustain large currents, but are not limited to these. Specifically, without limiting other possibilities, at least one electrical insulating material may be, or include, at least one insulating resin, such as insulating epoxy resin used in the manufacture of electronic printed circuit boards. In particular, the electrical insulating material may include, or may include, thermoplastic materials, polycarbonate, polyester such as polyethylene terephthalate (PET), polyvinyl chloride (PVC), polyurethane, polyether, polyamide, polyimide or copolymers thereof, such as glycol-modified polyethylene terephthalate, polyethylene naphthalate, polytetrafluoroethylene (PTFE) or alumina.

[0060] In the method and sample sensor according to the present invention, the sensor substrate may include two opposing sides, at least a first side and at least a second side facing the first side.

[0061] Specifically, the sample sensor, and more specifically the sensor substrate, may further include at least one additional electrode, the at least one of a reference electrode and a counter electrode. In one embodiment, the at least one additional electrode includes a pair / reference composite electrode. In particular, the reference electrode may include at least one reference electrode conductive material, and / or the counter electrode may include at least one counter electrode conductive material. More specifically, the at least one additional electrode may be located on at least one of a first side surface and a second side surface facing the first side surface of the sensor substrate.

[0062] As used herein, the term “conductive material” is a broad term and should be given its usual customary meaning to those skilled in the art, and should not be limited to any special or customized meaning. Specifically, the term may refer to, but not limited to, a conductive strip, layer, wire, or other kind of elongated conductor. More specifically, the term “conductive material” may refer to, but not limited to, a material that is conductive and therefore capable of sustaining an electric current, for example, a conductive material may include at least one material selected from the group consisting of carbon; carbon paste; gold; copper; silver; nickel; platinum; and palladium. Specifically, a conductive material may be, or include, one or more metals such as gold, copper, silver, nickel, palladium, or platinum. Additionally or alternatively, at least one conductive material may be, or include, at least one conductive compound, such as at least one conductive organic or inorganic compound. Additionally or alternatively, at least one conductive material may be at least one nonmetallic conductive material, such as polyaniline, poly-3,4-ethylenedioxythiophene (PEDOT), carbon, or carbon paste, or may contain these. Specifically, carbon paste may relate to a material comprising carbon, a solvent such as diethylene glycol butyl ether, and at least a binder such as vinyl chloride copolymer and ternary copolymer. Preferably, the conductive material according to the present invention may contain gold and / or carbon, and more preferably, the conductive material may consist of gold and / or carbon and / or carbon paste. Specifically, the conductive material may contain gold and further materials, such as carbon.

[0063] Furthermore, the conductive material may comprise at least one further layer of at least one additional material, specifically, the further layer may comprise a further conductive material. More specifically, the further layer of conductive material may comprise or consist of carbon. The further material may be arranged on the first side surface. The use of the further layer, particularly carbon, may contribute to efficient electron transfer by the conductive material.

[0064] The conductive material may have a thickness of at least about 0.1 μm, preferably at least about 0.5 μm, more preferably at least about 5 μm, specifically at least about 7 μm, or at least about 10 μm. If the conductive material contains or is carbon, the conductive material may have a thickness of at least about 7 μm, more specifically at least about 10 μm, for example, about 10 μm to 15 μm. Specifically, if the conductive material is gold, the conductive material may have a thickness of at least about 100 nm, more specifically at least about 500 nm.

[0065] The minimum thicknesses described above may be advantageous in ensuring proper electron transport. Thicknesses below the specified values ​​are generally insufficient for reliable electron transport. More specifically, they should not exceed approximately 30 μm for carbon and approximately 5 μm for gold. If the thickness is too large, the overall thickness and therefore the size of the sample sensor may increase. Larger sample sensor sizes are generally undesirable as they can cause difficulties during implantation. Furthermore, they may be less flexible, especially in the case of carbon, and / or expensive, especially in the case of gold.

[0066] The conductive material may also be hydrophobic. For example, the contact angle between the conductive material and water can be in the range of 60° to 140°, particularly about 100°, for a water droplet volume of, say, 5 μl, as determined by microscopy using a Keyence VHX-100.

[0067] The conductive material may further have a rough surface. A rough surface usually increases the efficiency of electron transfer. Furthermore, it increases hydrophobicity. A rough surface means that the surface may contain irregularities. The depth of these irregularities may be in the range of 1 μm to 6 μm, for example about 3 μm, as determined by, for example, an optical scanning microscope, especially a laser scanning microscope. The distance between two raised areas on the rough surface may be in the range of 20 μm to 80 μm, for example about 40 μm, as determined by, for example, an optical scanning microscope, especially a laser scanning microscope.

[0068] As used herein, the terms “reference electrode conductive material” and “counter electrode conductive material” are broad terms, given their ordinary and customary meanings to those skilled in the art, and should not be limited to any special or customized meanings. Specifically, these terms may refer to, but not limited to, conductive strips, layers, wires or other types of elongated electrical conductors present on the reference electrode or counter electrode, respectively. More specifically, these terms may refer to, but not limited to, materials that are conductive and therefore capable of sustaining an electric current, for example, the reference electrode conductive material and / or counter electrode conductive material may include at least one of the materials described herein with respect to conductive materials. In addition to the materials described above, the reference electrode conductive material and / or counter electrode conductive material may specifically include Ag / AgCl.

[0069] As used herein, the term “detection material” is a broad term and should be given its usual and customary meaning to those skilled in the art, and should not be limited to any special or customized meaning.

[0070] The detection material comprises at least one enzyme, specifically, the enzyme capable of catalyzing a chemical reaction that consumes at least the sample. Specifically, the enzyme is an H2O2-producing and / or consuming enzyme; more specifically, glucose oxidase (EC 1.1.3.4), hexose oxidase (EC 1.1.3.5), (S)-2-hydroxy acid oxidase (EC 1.1.3.15), cholesterol oxidase (EC 1.1.3.6), glucose dehydrogenase (EC 1.1.1.47), galactose oxidase (EC 1.1.3.9), alcohol oxidase (EC 1.1.3.13), L-glutamate oxidase (EC 1.4.3.11), or L-aspartate oxidase (EC 1.4.3.16); and more specifically, glucose oxidase (GOx) including any modifications thereof.

[0071] Furthermore, the detection material may also contain a crosslinking agent, such as a chemical crosslinking agent or a UV crosslinking agent, as described above.

[0072] Furthermore, the detection material may contain at least one polymer transition metal complex. The term “polymer transition metal complex” specifically refers to a material that may be at least one polymer material, or may contain at least one polymer material, and more specifically, it may be at least one polymer material and at least one metal-containing complex, or may contain both. The metal-containing complex may be selected from the group of transition metal element complexes, specifically, the metal-containing complex may be selected from osmium complexes, ruthenium complexes, vanadium complexes, cobalt complexes, and iron complexes, e.g., ferrocene, e.g., 2-aminoethylferrocene. More specifically, the detection material may contain polymer transition metal complexes, e.g., those described in International Publication No. 01 / 36660A2, and the content thereof is included by reference. In particular, the detection material may contain a modified poly(vinylpyridine) skeleton supporting a poly(vimidyl)Os complex covalently bonded via bidentate bonds. Suitable detection materials are further described in Feldmann et al, Diabetes Technology & Therapeutics, 5(5), 2003, 769-779, and are included by reference. Suitable detection materials may further include ferrocene-containing polyacrylamide-based viologen-modified redox polymers, pyrrole-2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)(ABTS)-pyrene, and naphthoquinone-LPEI. Polymer transition metal complexes may represent redox mediators incorporated into the crosslinked redox polymer network. This is advantageous because it can facilitate electron transfer between at least one enzyme or sample and the conductive material. To avoid sensor drift, the redox mediators and enzymes may be covalently incorporated into the polymer structure.

[0073] In certain embodiments, the detection material comprises at least an enzyme capable of catalyzing a chemical reaction that consumes a sample, particularly an H2O2-producing and / or consuming enzyme, a crosslinking agent, and a polymer transition metal complex. Specifically, the detection material may comprise at least a polymer transition metal complex and GOx, as well as a chemical crosslinking agent. More specifically, the detection material may comprise a modified poly(vinylpyridine) skeleton supporting a poly(biimididyl)Os complex, GOx, and a chemical crosslinking agent such as poly(ethylene glycol) diglycidyl ether (PEG-DGE) covalently bonded via bidentates. Further suitable detection materials are known to those skilled in the art.

[0074] In one embodiment, the detection material may include a polymer material and MnO2 particles.

[0075] The detection material according to the present invention may comprise, for example, about 40-60% by weight of a polymer transition metal complex, about 30-40% by weight of an enzyme capable of catalyzing a chemical reaction that consumes at least the sample, particularly an enzyme that produces and / or consumes H2O2, and about 0.5-25% by weight of a crosslinking agent based on the total dry weight of the detection material.

[0076] After coating the substrate, the detection material may be subjected to at least one curing step, in which at least a portion of the detection material is crosslinked. The curing step may be carried out as described above. Specifically, the curing step may be carried out after coating and before drying. Furthermore, the curing step may be carried out at least partially before any laser irradiation, or alternatively, after laser irradiation.

[0077] In certain embodiments, the sample sensor further includes at least one biocompatible membrane disposed on a flux-restricted polymer membrane. The term “biocompatible membrane” refers to a polymer membrane different from the polyvinylpyridine-based flux-restricted polymer membrane described above.

[0078] For example, the biocompatible membrane may be a gel membrane, which may be permeable to body fluids or at least the sample contained therein, and impermeable to the compound contained in the sample sensor, particularly the working electrode, thereby preventing its movement into body tissue.

[0079] The biocompatible layer may have a thickness of approximately 1 μm to approximately 10 μm, and in one embodiment, approximately 3 μm to approximately 6 μm. More specifically, the biocompatible layer covers the sample sensor at least partially or completely. Even more specifically, the biocompatible layer may be the outermost layer of the sample sensor. The biocompatible membrane layer may be or may contain the following materials: biodegradable polysaccharides such as methacrylate polymers and copolymers, acrylamide-methacrylate copolymers, hyaluronic acid (HA), agarose, dextran, chitosan, and poly(vinylpyridine) polymers. If the biocompatible membrane layer contains a poly(vinylpyridine) polymer, this poly(vinylpyridine) polymer may be the same as or different from the poly(vinylpyridine) polymer of the flux-limiting membrane. Preferably, it is different from the poly(vinylpyridine) polymer of the flux-limiting membrane.

[0080] The biocompatible film layer can be applied by techniques known to those skilled in the art, using at least one coating process, specifically a wet coating process, as described above.

[0081] In certain embodiments, the sample sensor does not include a biocompatible membrane placed on a flux-restricted polymer membrane. In these embodiments, the polyvinylpyridine polymer of the present invention may be the outermost layer of the sample sensor. Therefore, the flux-restricted polymer membrane can also function as a biocompatible membrane.

[0082] The present invention further relates to a polymer membrane for use in a sample sensor, wherein the polymer membrane is of formula (I): TIFF0007871259000003.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. It contains a poly(vinylpyridine) polymer having the following properties.

[0083] Preferably, the characteristics of the polymer film are as described above for the flux-restricted polymer film.

[0084] The present invention further includes the following formula (I): TIFF0007871259000004.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. A polyvinylpyridine polymer having, Crosslinking agent and solvent and The present invention relates to a liquid composition suitable for coating a flux-restricted polymer film onto a sample sensor, which includes [a specific component].

[0085] Preferably, the polyvinylpyridine polymer and the crosslinking agent have the characteristics described above.

[0086] The solvent may include a non-aqueous organic solvent, particularly a water-miscible solvent, such as methanol, ethanol, propanol, or any combination thereof, or a mixture of a non-aqueous organic solvent and water. Preferably, the non-aqueous organic solvent is ethanol. Preferably, the composition contains about 50% to about 90% (v / v) of a non-aqueous organic solvent, such as ethanol, and about 10% to about 50% (v / v) of water, preferably about 70% to about 90% (v / v) of a non-aqueous organic solvent, and about 10% to about 30% (v / v) of water, particularly about 80% (v / v) of a non-aqueous organic solvent, such as ethanol, and about 20% (v / v) of water. In further embodiments, the solvent consists of a non-aqueous solvent, particularly ethanol.

[0087] In certain embodiments, the liquid composition may contain about 100 mg / ml to about 140 mg / ml of polymer and about 8 mg / ml to about 12 mg / ml of crosslinking agent, particularly about 120 mg / ml of polymer and about 10 mg / ml of crosslinking agent. The weight ratio of polymer to crosslinking agent is preferably about 12:1.

[0088] The present invention further relates to a method for manufacturing a sample sensor, a) A step of providing a substrate having a first side surface and a second side surface, and at least one working electrode disposed on the first side surface of the substrate, b) A step of forming a flux-restricting polymer film on at least one working electrode, wherein the flux-restricting polymer film is defined by the following formula (I): TIFF0007871259000005.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. The present invention relates to a method comprising a poly(vinylpyridine) polymer having the following properties.

[0089] Preferably, the characteristics of the polymer film are as described above.

[0090] In certain embodiments, step b) includes applying a liquid composition comprising a polymer, a crosslinking agent, and a solvent onto a working electrode. In particular, step b) includes coating at least one working electrode with a liquid composition comprising a polymer, a crosslinking agent, and a solvent, curing the polymer, and drying it. Preferably, the characteristics of the liquid composition, the application step, the curing step, and the drying step are as described above.

[0091] In certain embodiments of the method according to the present invention, at least a second film layer may be applied in addition to at least one flux-restricted polymer film layer. The second film layer may be a biocompatible film layer, preferably the biocompatible film layer described above.

[0092] The method according to the present invention may further include at least one diffusion step, in which the crosslinking agent contained in the flux-restricted polymer film can be at least partially diffused into the detection material. Diffusion may occur while the film layer is applied to the detection material. Diffusion of the crosslinking agent into the detection material may enable at least partial crosslinking of the detection material when the detection material is applied to the substrate, regardless of the amount of crosslinking agent in the detection material.

[0093] The diffusion step may further include swelling of at least a portion of the detection material. As used herein, the term “swelling” is a broad term, and its usual, customary meaning should be given to those skilled in the art, and should not be limited to any special or customized meaning. Specifically, the term may refer, but not limited to, the binding of water and / or water-soluble solvents such as ethanol, methanol, or acetone to the material, more specifically, the binding of water and / or water-soluble solvents to the detection material. The uptake of water and / or water-soluble solvents into the detection material may favorably enable the diffusion of the crosslinking agent into the detection material, which may be necessary for efficient crosslinking. Swelling may further refer to the uptake of water from the membrane layer.

[0094] In order to enable sufficient swelling in the method according to the present invention, the polymer material in the sensing material may be able to absorb at least 10% by weight, more specifically at least 20% by weight, even more specifically at least 30% by weight, and even more specifically up to 90% by weight of water and / or solvent from the film layer, based on the dry weight of the polymer material, within a time frame of several minutes, for example, 1 to 15 minutes.

[0095] This swelling and / or incorporation of water and / or solvent is advantageous because it may thereby allow the diffusion of the crosslinking agent from the film layer to the sensing material.

[0096] Furthermore, the present invention relates to the use of the above-described sample sensor for detecting at least one sample in a sample, specifically in a sample of body fluids. More specifically, the sample sensor is a sensor for continuous glucose measurement.

[0097] As used herein, the term “body fluid” refers to all body fluids of the subject known to or suspected to contain the specimen of the present invention, including interstitial fluid, blood, plasma, tears, urine, lymph, cerebrospinal fluid, bile, feces, sweat, and saliva. In general, any type of body fluid may be used. Preferably, the body fluid is a fluid present in the user’s body tissue, such as interstitial tissue. Thus, as an example, the body fluid may be selected from the group consisting of blood and interstitial fluid. However, one or more other types of body fluid may be used additionally or alternatively. Body fluids can generally be contained in body tissue. Thus, generally, the detection of at least one specimen in a body fluid can be determined, preferably in vivo.

[0098] The term "sample" will be understood by those skilled in the art and refers to any lower part of a body fluid. Samples can be obtained by well-known techniques, including, for example, vein or arterial puncture, epidermal puncture, etc.

[0099] The terms “User” and “Subject” are used interchangeably herein and are broad terms, to be given the ordinary and customary meanings to those skilled in the art, and should not be limited to any special or customized meanings. Specifically, these terms may refer to, but are not limited to, a human or an animal, regardless of whether they may be in a healthy state or may be suffering from one or more diseases, respectively. For example, the subject may be a human or an animal suffering from diabetes. However, additionally or alternatively, the present invention may be applied to other types of subjects.

[0100] Furthermore, the present invention relates to a method for measuring a sample in a sample including the sample sensor described herein.

[0101] The method for measuring specimens according to the present invention may, in particular, be an in vivo method. Alternatively, the method of the present invention may also include the measurement of specimens in samples of bodily fluids obtained, for example, from subjects, particularly human subjects, under in vitro conditions. Specifically, the method does not necessarily include the diagnosis of a disease based on the measurement.

[0102] Further aspects of the present invention are given by the following formula (Ia): TIFF0007871259000006.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, (z is approximately 0 to approximately 20 mol%) Regarding polymers having the following characteristics: The polymer is (i) Statistical polymers; (ii) Polymers having a weight-average molecular weight of approximately 60 kDa to approximately 200 kDa, and / or (iii) A polymer that is crosslinked with a crosslinking agent which is N,N-diglycidyl-4-glycidyloxyaniline, and in particular, the weight ratio of polymer to crosslinking agent is about 8:1 (w / w) to about 16:1 (w / w), about 10:1 (w / w) to about 14:1 (w / w), and in particular about 12:1 (w / w).

[0103] Further embodiments include a sample sensor according to this specification comprising the polymer of formula (Ia), a polymer membrane according to this specification comprising the polymer of formula (Ia), a liquid composition according to this specification comprising the polymer of formula (Ia), and a method for producing a sample sensor according to this specification, comprising using the polymer of formula (Ia).

[0104] The preferred characteristics of the sample sensor, polymer membrane, liquid composition, and manufacturing method are as described herein for the polymer of formula (I).

[0105] Further optional features and embodiments are disclosed in more detail in subsequent descriptions of embodiments, preferably in conjunction with dependent claims. Herein, each optional feature may be implemented independently and in any viable combination, as will be understood by those skilled in the art. The scope of the present invention is not limited by preferred embodiments.

[0106] In summary, without excluding further possible embodiments, the following embodiments can be envisioned.

[0107] 1. A sample sensor comprising at least one working electrode and a flux-restricting polymer membrane disposed on the at least one working electrode, wherein the flux-restricting polymer membrane comprises the following formula (I): TIFF0007871259000007.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. A sample sensor containing a polymer having the properties of a polymer.

[0108] 2.x is approximately 2 to 8 mol%, y is approximately 82 to 88 mol%, A sample sensor as described in item 1, wherein z is approximately 7 to approximately 13 mol%.

[0109] 3.x is approximately 3 to 7 mol%, y is approximately 83 to 87 mol%, A sample sensor according to item 1 or 2, wherein z is approximately 8 to approximately 12 mol%.

[0110] 4.x is approximately 4 to 6 mol%, y is approximately 84 to 86 mol%, A sample sensor according to any one of items 1 to 3, wherein z is approximately 9 to approximately 11 mol%.

[0111] 5.x is approximately 5 mol%, y is approximately 85 mol%, A sample sensor according to any one of items 1 to 4, wherein z is approximately 10 mol%.

[0112] 6. A sample sensor according to any one of items 1 to 5, wherein the polymer is a statistical copolymer.

[0113] 7. The sample sensor according to any one of claims 1 to 6, wherein the polymer has a weight-average molecular weight of approximately 60 kDa to approximately 200 kDa, approximately 80 kDa to approximately 160 kDa, approximately 100 kDa to approximately 140 kDa, and particularly approximately 120 kDa.

[0114] 8. The sample sensor according to any one of claims 1 to 7, wherein the polymer has a number average molecular weight of about 4 kDa to about 90 kDa, about 40 kDa to about 70 kDa, and particularly about 60 kDa.

[0115] 9. The sample sensor according to any one of claims 1 to 8, wherein the polymer has a polydispersity index of about 1.4 to about 3, particularly about 2.

[0116] 10. A sample sensor according to any one of items 1 to 9, wherein the polymer of formula (I) in the flux-restricting polymer film is crosslinked with a crosslinking agent.

[0117] 11. The sample sensor according to item 10, wherein the weight ratio of polymer to crosslinking agent in the flux-restricting polymer membrane is approximately 8:1 (w / w) to approximately 16:1 (w / w), approximately 10:1 (w / w) to approximately 14:1 (w / w), and particularly approximately 12:1 (w / w).

[0118] 12. The sample sensor according to item 10 or 11, wherein the crosslinking agent comprises at least one functional epoxide group.

[0119] 13. The sample sensor according to any one of claims 10 to 12, wherein the crosslinking agent comprises at least two, particularly three, functional epoxide groups.

[0120] 14. The sample sensor according to any one of items 10 to 13, wherein the crosslinking agent is N,N-diglycidyl-4-glycidyloxyaniline.

[0121] 15. (i) A substrate, The first aspect and the second aspect, At least one conductive material disposed on the first side surface of the substrate and A substrate comprising, (ii) A working electrode comprising a sensing material that at least partially covers a first side surface of a substrate, wherein the sensing material comprises at least one enzyme, (iii) A flux-restricting polymer film disposed on the at least one working electrode, wherein the flux-restricting polymer film comprises a polymer of formula (I), A sample sensor as described in any one of items 1 to 14, including the following:

[0122] 16. The sample sensor, (i) A substrate, The first aspect and the second aspect, At least one conductive material disposed on the first side surface of the substrate and A substrate comprising, (ii) The working electrode comprising a sensing material, wherein the working electrode at least partially covers the first side surface of the substrate and is at least partially disposed on the at least one conductive material, Equipped with, A sample sensor according to any one of claims 1 to 14, wherein the detection material comprises at least one enzyme.

[0123] 17. The specimen sensor according to item 15 or 16, wherein the at least one conductive material disposed on the first side surface of the substrate is selected from gold, carbon, carbon paste and any combination thereof.

[0124] 18. A sample sensor according to any one of items 15 to 17, wherein the detection material comprises the enzyme glucose oxidase (GOx).

[0125] 19. A sample sensor according to any one of claims 15 to 18, wherein the detection material further comprises at least one crosslinking agent.

[0126] 20. The sample sensor according to any one of claims 15 to 19, wherein the detection material further comprises at least one polymer metal-containing complex.

[0127] 21. The sample sensor according to item 20, wherein the at least one polymer metal-containing complex is selected from the group of polymer transition metal-containing complexes.

[0128] 22. The sample sensor according to item 21, wherein the at least one polymer transition metal-containing complex is selected from osmium complexes, ruthenium complexes, vanadium complexes, cobalt complexes, and iron complexes.

[0129] 23. A sample sensor according to any one of items 1 to 22, comprising at least one additional electrode.

[0130] 24. The specimen sensor according to item 23, wherein the at least one further electrode is selected from a counter electrode, a reference electrode, and a pair / reference composite electrode.

[0131] 25. The specimen sensor according to item 23 or 24, wherein the at least one further electrode is a pair / reference composite electrode.

[0132] 26. A specimen sensor according to any one of items 1 to 25, which is a two-electrode sensor comprising one working electrode and one pair / reference composite electrode.

[0133] 27. A sample sensor according to any one of claims 1 to 26, further comprising at least one biocompatible membrane disposed on the flux-restricting polymer membrane.

[0134] 28. A sample sensor according to any one of claims 1 to 27, which does not include a biocompatible membrane disposed on the flux-restricting polymer membrane.

[0135] 29. The sample sensor according to any one of items 1 to 28, wherein the flux-restricting polymer membrane is glucose permeable.

[0136] 30. The flux-restricted polymer film is at least about 1 × 10 -6 cm 2 / s, preferably about 1 × 10 -6 cm 2 / s~approx. 1×10 -10 cm 2 A sample sensor according to any one of items 1 to 29, having a glucose diffusion coefficient of / s.

[0137] 31. Use of a sample sensor as described in any one of items 1 to 30 for detecting at least one sample in a sample.

[0138] 32. A method for determining a sample in a sample, comprising using a sample sensor described in any one of items 1 to 30.

[0139] 33. A polymer film for use in a sample sensor, wherein the polymer film comprises the following formula (I): TIFF0007871259000008.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. A polymer film containing a polymer having the following properties.

[0140] 34. A polymer film according to item 33, comprising at least one feature defined in any one of items 2 to 15.

[0141] 35. A liquid composition, the following formula (I): TIFF0007871259000009.tif4554(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, A polymer having z (0 to approximately 20 mol%), Crosslinking agent and solvent and A liquid composition containing the following:

[0142] 36. The liquid composition according to item 35, comprising at least one feature defined in any one of items 2 to 15.

[0143] 37. The liquid composition according to item 35 or 36, wherein the solvent comprises ethanol and water, particularly about 70% to about 90% (v / v) ethanol and about 10% to about 30% (v / v) water, particularly about 80% (v / v) ethanol and about 20% (v / v) water.

[0144] 38. A liquid composition according to any one of claims 35 to 37, comprising about 100 mg / ml to about 140 mg / ml of polymer and about 8 mg / ml to about 12 mg / ml of crosslinking agent, particularly about 120 mg / ml of polymer and about 10 mg / ml of crosslinking agent.

[0145] 39. A sample sensor, in particular a method for manufacturing a sample sensor according to any one of items 1 to 30, a) A step of providing a substrate having a first side surface and a second side surface, and at least one working electrode disposed on the first side surface of the substrate, b) A step of forming a polymer film layer on at least one working electrode, wherein the polymer film is of the following formula (I): TIFF0007871259000010.tif4560(In the formula, x is approximately 2 to approximately 8 mol%, y is approximately 72 to 98 mol%, z is between 0 and approximately 20 mol%. A method comprising a polymer having the following properties.

[0146] 40. The method described in paragraph 39, comprising at least one feature defined in any one of paragraphs 2 to 15.

[0147] 41. The method according to claim 39 or 40, wherein step b) comprises coating at least one working electrode with a liquid composition comprising a polymer, a crosslinking agent, and a solvent, curing the polymer, and drying.

[0148] 42. The method according to claim 41, wherein the solvent comprises ethanol and water, in particular about 70% to about 90% (v / v) ethanol and about 10% to about 30% (v / v) water, in particular about 80% (v / v) ethanol and about 20% (v / v) water.

[0149] 43. The method according to item 41 or 42, wherein the liquid composition comprises about 100 mg / ml to about 140 mg / ml of polymer and about 8 mg / ml to about 12 mg / ml of crosslinking agent, particularly about 120 mg / ml of polymer and about 10 mg / ml of crosslinking agent.

[0150] 44. A sample sensor comprising at least one working electrode and a flux-restricting polymer membrane disposed on the at least one working electrode, wherein the flux-restricting polymer membrane is provided for the following formula (Ia): TIFF0007871259000011.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, z is approximately 0 to 20 mol%. A sample sensor comprising a statistical polymer having the following properties.

[0151] 45. A sample sensor according to item 44, comprising at least one feature defined in any one of items 1 to 30.

[0152] 46. ​​A polymer membrane for use in a sample sensor, wherein the polymer membrane comprises the following formula (Ia): TIFF0007871259000012.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, z is approximately 0 to 20 mol%. A polymer film comprising a statistical polymer having the properties of a polymer.

[0153] 47. A polymer film according to clause 46, comprising at least one feature defined in any one of clauses 33 to 34.

[0154] 48. A liquid composition, the following formula (Ia): TIFF0007871259000013.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, z is approximately 0 to 20 mol%. A statistical polymer having, Crosslinking agent and solvent and A liquid composition containing the following:

[0155] 49. The liquid composition according to item 48, comprising at least one feature defined in any one of items 35 to 38.

[0156] 50. A sample sensor, in particular a method for manufacturing a sample sensor as described in any one of items 44 to 45, a) A step of providing a substrate having a first side surface and a second side surface, and at least one working electrode disposed on the first side surface of the substrate, b) A step of forming a polymer film layer on at least one working electrode, wherein the polymer film is defined by the following formula (Ia): TIFF0007871259000014.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, z is approximately 0 to 20 mol%. A method comprising a statistical polymer having the properties of a statistical polymer.

[0157] 51. The method described in paragraph 50, comprising at least one feature defined in any one of paragraphs 35 to 38.

[0158] 52. A sample sensor comprising at least one working electrode and a flux-restricting polymer membrane disposed on the at least one working electrode, wherein the flux-restricting polymer membrane comprises a polymer having a weight-average molecular weight of about 60 kDa to about 200 kDa, and the polymer is given by the following formula (Ia): TIFF0007871259000015.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, z is approximately 0 to 20 mol%. A specimen sensor having the following features.

[0159] 53. A sample sensor according to item 52, comprising at least one feature defined in any one of items 1 to 30.

[0160] 54. The polymer film has a weight-average molecular weight of approximately 60 kDa to approximately 200 kDa, and is given by the following formula (Ia): TIFF0007871259000016.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, z is approximately 0 to 20 mol%. A polymer membrane for use in a sample sensor, comprising a polymer having the properties of a polymer.

[0161] 55. A polymer film according to clause 54, comprising at least one feature defined in any one of clauses 33 to 34.

[0162] 56. Having a weight-average molecular weight of approximately 60 kDa to approximately 200 kDa, as shown in formula (Ia): TIFF0007871259000017.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, A polymer having z (approximately 0 to approximately 20 mol%), Crosslinking agent and solvent and A liquid composition containing the following:

[0163] 57. The liquid composition according to item 56, comprising at least one feature defined in any one of items 35 to 38.

[0164] 58. A sample sensor, in particular a method for manufacturing a sample sensor as described in any one of items 52 to 53, a) A step of providing a substrate having a first side surface and a second side surface, and at least one working electrode disposed on the first side surface of the substrate, b) A step of forming a polymer film layer on at least one working electrode, wherein the polymer film has a weight-average molecular weight of about 60 kDa to about 200 kDa, and the following formula (Ia): TIFF0007871259000018.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, z is approximately 0 to 20 mol%. A process, a method, comprising a polymer having a polymer.

[0165] 59. The method described in paragraph 58, comprising at least one feature defined in any one of paragraphs 35-38.

[0166] 60. A sample sensor comprising at least one working electrode and a flux-restricting polymer film disposed on the at least one working electrode, wherein the flux-restricting polymer film comprises the following formula (Ia): TIFF0007871259000019.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, The polymer contains z (approximately 0 to approximately 20 mol%) A sample sensor in which the polymer is crosslinked with a crosslinking agent, N,N-diglycidyl-4-glycidyloxyaniline, and in particular, the weight ratio of the polymer to the crosslinking agent is about 8:1 (w / w) to about 16:1 (w / w), about 10:1 (w / w) to about 14:1 (w / w), and especially about 12:1 (w / w).

[0167] 61. A sample sensor according to item 60, comprising at least one feature defined in any one of items 1 to 30.

[0168] 62. A polymer film for use in a sample sensor, wherein the polymer film comprises the following formula (Ia): TIFF0007871259000020.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, The polymer contains z (approximately 0 to approximately 20 mol%) A polymer film in which the polymer is crosslinked with a crosslinking agent which is N,N-diglycidyl-4-glycidyloxyaniline, and in particular the weight ratio of polymer to crosslinking agent is about 8:1 (w / w) to about 16:1 (w / w), about 10:1 (w / w) to about 14:1 (w / w), and in particular about 12:1 (w / w).

[0169] 63. The method described in paragraph 62, comprising at least one feature defined in any one of paragraphs 33 to 34.

[0170] 64. A liquid composition, the following formula (Ia): TIFF0007871259000021.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, The polymer contains z (approximately 0 to approximately 20 mol%) Crosslinking agent and A solvent is included, A method wherein the crosslinking agent is N,N-diglycidyl-4-glycidyloxyaniline, and in particular, the weight ratio of polymer to crosslinking agent is about 8:1 (w / w) to about 16:1 (w / w), about 10:1 (w / w) to about 14:1 (w / w), and in particular about 12:1 (w / w).

[0171] 65. The liquid composition according to claim 64, comprising at least one feature defined in any one of claims 35 to 38.

[0172] 66. A sample sensor, in particular a method for manufacturing a sample sensor as described in any one of items 60 to 61, a) A step of providing a substrate having a first side surface and a second side surface, and at least one working electrode disposed on the first side surface of the substrate, b) A step of forming a polymer film layer on at least one working electrode, wherein the polymer film is defined by the following formula (Ia): TIFF0007871259000022.tif4560(In the formula, x is approximately 2 to approximately 20 mol%, y is approximately 60 to 98 mol%, The process includes a polymer having z (where z is approximately 0 to approximately 20 mol%), Step (b) comprises crosslinking the polymer with a crosslinking agent which is N,N-diglycidyl-4-glycidyloxyaniline, in particular the weight ratio of polymer to crosslinking agent which is about 8:1 (w / w) to about 16:1 (w / w), about 10:1 (w / w) to about 14:1 (w / w), and in particular about 12:1 (w / w), in a method.

[0173] 67. The method described in paragraph 66, comprising at least one feature defined in any one of paragraphs 35-38. [Brief explanation of the drawing]

[0174] [Figure 1] Figure 1 shows the run-in currents of six different sample sensors. [Figure 2] Figure 2 shows the currents of six different sample sensors over approximately 10 days. [Figure 3] Figure 3 shows the long-term changes in sensitivity of sample sensors containing different flux-limiting polymer membranes at a glucose concentration of 180 mg / dL.

[0175] The present invention is not limited to any of the embodiments described above and can be modified in a wide variety of ways. Those skilled in the art will recognize that embodiments of the present invention can be easily adapted without departing from the scope of the invention. Thus, simple adaptations can be conceived for the preparation of sample sensors. The present invention enables the preparation of samples with reproducible sensor sensitivity while reducing production costs. Further features, details, and advantages of the present invention are derived from the following description of embodiments based on the claims and drawings.

[0176] The contents of all references cited in this patent application are incorporated herein by reference to their respective specific disclosures and their entirety. [Examples]

[0177] The following examples serve to illustrate the present invention. They should not be construed as limiting the scope of protection.

[0178] Example 1: Preparation and testing of sample sensors containing different flux-limiting polymer film layers Sensor substrates based on polyethylene terephthalate and gold thin layers were coated with carbon paste using the doctor blade method. Suitable carbon conductive inks are available from Ercon, Inc. (Wareham, Massachusetts), EIdu Pont de Nemours and Co. (Wilmington, Delaware), Emca-Remex Products (Montgomeryville, Pennsylvania), or TEKRA, A Division of EIS, Inc. (New Berlin, Wisconsin). The carbon paste was then dried at 50°C for 12 hours.

[0179] A layer of detection material was applied to the sensor substrate by cannula coating, and then dried at ambient temperature, for example, about 25°C, for 10 minutes.

[0180] The detection material in each case contained 57 wt% polymer transition metal complex (modified poly(vinylpyridine) skeleton supporting a poly(vimidyl)Os complex covalently bonded via bidentate), 33 wt% glucose oxidase, and 10 wt% PEG-DGE (poly(ethylene glycol)-diglycidyl ether), based on the total weight percentages of the polymer transition metal complex, glucose oxidase, and PEG-DGE. Water was used as the solvent. The total concentration of the polymer transition metal complex, glucose oxidase, and PEG-DGE in water was 50 mg / ml.

[0181] A working electrode with dimensions of 0.5 mm × 0.6 mm and a layer thickness of 3 μm was prepared on the sensor substrate by laser ablation.

[0182] As shown in Table 1, the working electrode of each sensor was coated with a different liquid composition containing a poly(vinylpyridine) polymer and a crosslinking agent. In all cases, a mixture of 80% (v / v) ethanol and 20% (v / v) water was used as the solvent. In the table, Oxi-Ani refers to N,N-diglycidyl-4-glycidyloxyaniline, and PEG-DGE 200 refers to poly(ethylene glycol)-diglycidyl ether with a number-average molecular weight of 200 Da.

[0183] [Table 1]

[0184] After coating, the sensor was dried, and the polymer was crosslinked at room temperature to obtain a flux-limiting film on the working electrode. Silver / silver chloride was used as the pair / reference composite electrode.

[0185] Four sensors were prepared for each of the different flux-limiting membranes. The measurements described below were performed for each of the four sensors, and the median of the obtained values ​​was calculated. All measurements were performed in vitro. All descriptions below refer to the median values ​​of the measured values.

[0186] For all sensors, current was measured over approximately 10 days using various amounts of glucose. Measurements were performed in phosphate buffer. After a run-in period of approximately 6 hours, glucose was added. The glucose was periodically replaced by flushing with phosphate buffer and phosphate-glucose buffer in a fixed ratio for each step using an LC system (Jasco LC-4000 series). Each step had a duration of 90 minutes in the pyramidal phase. The following glucose concentrations were used: 0 mg / dl, 36 mg / dl, 72 mg / dl, 108 mg / dl, 144 mg / dl, 216 mg / dl, 306 mg / dl, 414 mg / dl, 468 mg / dl, 360 mg / dl, 270 mg / dl, 180 mg / dl, 126 mg / dl, 90 mg / dl, 54 mg / dl, and 14.4 mg / dl. Sensitivity was expressed at a glucose concentration of 180 mg / dl.

[0187] Figure 1 shows the median run-in time t in minutes for different sensors. Sensors containing poly(ethylene glycol)-diglycidyl ether as a crosslinking agent exhibit shorter run-in times compared to sensors containing N,N-diglycidyl-4-glycidyloxyaniline as a crosslinking agent. However, these run-in times are still sufficiently short to enable use as continuous glucose sensors.

[0188] Figure 2 shows the current I over time t for different sensors, measured in minutes. The sensor containing 10 mol% sulfonate-vinylpyridinium in a poly(vinylpyridine) polymer shows a rapid decrease in current over time, while the sensor containing 5 mol% sulfonate-vinylpyridinium maintains a nearly constant current throughout the measurement period.

[0189] Therefore, sensors B3, B5, and B6 exhibit significantly increased long-term stability compared to sensors V1, V2, and V4.

[0190] Figure 3 shows the sensitivity of different sensors over time t in days, expressed in nA / mg / dl. It can be seen that the sensitivity degradation of sensors B3, B5, and B6 is lower than that of sensors V1, V2, and V4. This is advantageous as it indicates higher sensor stability over longer periods.

[0191] Example 2: Sample sensor containing different flux-limiting polymer film layers A polyethylene terephthalate-based sensor substrate was coated with carbon paste using the doctor blade method. Suitable carbon conductive inks are available from Ercon, Inc. (Wareham, Massachusetts), EIdu Pont de Nemours and Co. (Wilmington, Delaware), Emca-Remex Products (Montgomeryville, Pennsylvania), or TEKRA, A Division of EIS, Inc. (New Berlin, Wisconsin). The carbon paste was then dried at 50°C for 12 hours.

[0192] A layer of detection material was applied to the sensor substrate by cannula coating, and then dried at ambient temperature, for example, about 25°C, for 10 minutes.

[0193] The detection material in each case contained 57 wt% polymer transition metal complex (modified poly(vinylpyridine) skeleton supporting a poly(vimidyl)Os complex covalently bonded via bidentate), 33 wt% glucose oxidase, and 10 wt% PEG-DGE (poly(ethylene glycol)-diglycidyl ether), based on the total weight percentages of the polymer transition metal complex, glucose oxidase, and PEG-DGE. Water was used as the solvent. The total concentration of the polymer transition metal complex, glucose oxidase, and PEG-DGE in water was 50 mg / ml.

[0194] A working electrode with dimensions of 0.5 mm × 0.6 mm and a layer thickness of 4 μm was prepared on the sensor substrate by laser ablation.

[0195] As shown in Table 2, the working electrode of each sensor was coated with a different liquid composition containing a poly(vinylpyridine) polymer and a crosslinking agent (immersion coating, 3 times). In all cases, a mixture of 80% (v / v) ethanol and 20% (v / v) water was used as the solvent. In the table, Oxi-Ani means N,N-diglycidyl-4-glycidyloxyaniline, and PEG-DGE 200 means poly(ethylene glycol)-diglycidyl ether having a number-average molecular weight of 200 Da.

[0196] Sensors V1, V2, and B3 in Table 2 were prepared in the same manner as in Example 1, and V1, V2, and B3 in Table 2 correspond to V1, V2, and B3 in Table 1.

[0197] In Table 2, Mn is the number-average molecular weight of the polyvinylpyridine polymer. PDI is the polydispersity index of the polyvinylpyridine polymer.

[0198] [Table 2]

[0199] After coating, the sensor was dried to obtain a flux-limiting film on the working electrode. If a crosslinking agent was present, the polymer was crosslinked at room temperature.

[0200] Silver / silver chloride was used as the pair / reference composite electrode.

[0201] Four sensors were prepared for each of the different flux-limiting membranes. The measurements described below were performed for each of the four sensors, and the median of the obtained values ​​was calculated. All measurements were performed in vitro. All descriptions below refer to the median values ​​of the measured values.

[0202] For all sensors, current was measured over approximately 7 days using varying amounts of glucose. Measurements were performed in phosphate buffer.

[0203] Sensitivity was measured on the first day at a glucose concentration of 180 mg / dl. Average drift was calculated at glucose concentrations of 180 mg / dl and 468 mg / dl.

[0204] As can be seen from Table 2, the sensors containing the flux-limiting film of the present invention exhibit reduced drift compared to the comparative sensor, but still have a sufficiently short run-in time. At the same time, they still have good sensitivity.

Claims

1. A sample sensor comprising at least one working electrode and a flux-restricting polymer film disposed on the at least one working electrode, wherein the flux-restricting polymer film comprises the following formula (I): (In the formula, x is between 4 and 6 mol%, y is 82-88 mol%, z is 8 to 12 mol%, and (The sum of the mole percentages of x, y, and z is 100 mol%) A sample sensor containing a polymer having the properties of a polymer.

2. The sample sensor according to claim 1, wherein the polymer is a statistical copolymer.

3. The sensor according to claim 1 or 2, wherein the polymer has a weight-average molecular weight in the range of 60 kDa to 200 kDa.

4. The sample sensor according to any one of claims 1 to 3, wherein the polymer of formula (I) in the flux-limiting polymer film is crosslinked with a crosslinking agent.

5. The sample sensor according to claim 4, wherein the weight ratio of polymer to crosslinking agent in the flux-limiting polymer film is 8:1 (w / w) to 16:1 (w / w).

6. The sample sensor according to claim 5, wherein the weight ratio of polymer to crosslinking agent in the flux-limiting polymer film is 10:1 (w / w) to 14:1 (w / w).

7. The sample sensor according to claim 6, wherein the weight ratio of polymer to crosslinking agent in the flux-restricting polymer film is 12:1 (w / w).

8. The sample sensor according to any one of claims 4 to 7, wherein the crosslinking agent is N,N-diglycidyl-4-glycidyloxyaniline.

9. The aforementioned sample sensor, (i) A substrate, The first aspect and the second aspect, At least one conductive material disposed on the first side surface of the substrate, A substrate comprising, (ii) The working electrode comprising a sensing material, wherein the working electrode at least partially covers the first side surface of the substrate and is at least partially disposed on the at least one conductive material, Equipped with, The sample sensor according to any one of claims 1 to 8, wherein the detection material comprises at least one enzyme.

10. The at least one conductive material disposed on the first side surface of the substrate is selected from gold, carbon, carbon paste and any combination thereof, and / or The sample sensor according to claim 9, wherein the detection material comprises the enzyme glucose oxidase (GOx).

11. The specimen sensor according to any one of claims 1 to 10, which is a two-electrode sensor including the working electrode and a pair / reference composite electrode.

12. The method further includes at least one biocompatible membrane disposed on the flux-restricting polymer membrane, or A sample sensor according to any one of claims 1 to 11, wherein it does not include a biocompatible membrane disposed on the flux-restricting polymer membrane.

13. Use of a sample sensor according to any one of claims 1 to 12 for detecting at least one sample in a sample.

14. A method for determining a sample in a sample, comprising using a sample sensor according to any one of claims 1 to 12.

15. A polymer film for use in a sample sensor, wherein the polymer film comprises the following formula (I): (In the formula, x is between 4 and 6 mol%, y is 82-88 mol%, z is 8 to 12 mol%, and (The sum of the mole percentages of x, y, and z is 100 mol%) A polymer film containing a polymer having the properties of a polymer.

16. A liquid composition, the following formula (I): (In the formula, x is between 4 and 6 mol%, y is 82-88 mol%, z is 8 to 12 mol%, and A polymer having a total molar percentage of x, y, and z of 100 mol%, Crosslinking agent, and solvent, A liquid composition containing the following:

17. The liquid composition according to claim 16, wherein the solvent comprises ethanol and water.

18. The liquid composition according to claim 17, wherein the solvent comprises 70% to 90% (v / v) ethanol and 10% to 30% (v / v) water.

19. The liquid composition according to claim 18, wherein the solvent comprises 80% (v / v) ethanol and 20% (v / v) water.

20. A liquid composition according to any one of claims 16 to 19, comprising 100 mg / ml to 140 mg / ml of polymer and 8 mg / ml to 12 mg / ml of crosslinking agent.

21. The liquid composition according to claim 20, comprising 120 mg / ml of polymer and 10 mg / ml of crosslinking agent.

22. A method for manufacturing a sample sensor according to any one of claims 1 to 12, a) A step of providing a substrate having a first side surface and a second side surface, and at least one working electrode disposed on the first side surface of the substrate, b) A step of forming a flux-restricting polymer film layer on at least one working electrode, wherein the flux-restricting polymer film comprises the following formula (I): (In the formula, x is between 4 and 6 mol%, y is 82-88 mol%, z is 8 to 12 mol%, and (The sum of the mole percentages of x, y, and z is 100 mol%) A process comprising a polymer having, A method that includes this.

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