Patent Description:
Embodiments of the subject matter described herein relate generally to sensors for monitoring analyte levels in patients and to methods for fabricated such sensors. More particularly, embodiments of the subject matter relate to glucose sensors, such as for monitoring blood glucose level continuously, or substantially continuously.

The pancreas of a normal healthy person produces and releases insulin into the blood stream in response to elevated blood plasma glucose levels. Beta cells (β-cells), which reside in the pancreas, produce and secrete insulin into the blood stream as it is needed. If β-cells become incapacitated or die, a condition known as Type <NUM> diabetes mellitus (or in some cases, if β-cells produce insufficient quantities of insulin, a condition known as Type <NUM> diabetes), then insulin may be provided to a body from another source to maintain life or health.

Traditionally, because insulin cannot be taken orally, insulin has been injected with a syringe. More recently, the use of infusion pump therapy has been increasing in a number of medical situations, including for delivering insulin to diabetic individuals. For example, external infusion pumps may be worn on a belt, in a pocket, or the like, and they can deliver insulin into a body via an infusion tube with a percutaneous needle or a cannula placed in subcutaneous tissue.

As of <NUM>, less than <NUM>% of Type <NUM> diabetic individuals in the United States were using infusion pump therapy. Currently, over <NUM>% of the more than <NUM>,<NUM> Type <NUM> diabetic individuals in the U. are using infusion pump therapy. The percentage of Type <NUM> diabetic individuals that use an infusion pump is growing at a rate of over <NUM>% each year. Moreover, the number of Type <NUM> diabetic individuals is growing at <NUM>% or more per year, and growing numbers of insulin-using Type <NUM> diabetic individuals are also adopting infusion pumps. Additionally, physicians have recognized that continuous infusion can provide greater control of a diabetic individual's condition, so they too are increasingly prescribing it for patients.

An infusion pump system may include an infusion pump that is automatically and/or semi-automatically controlled to infuse insulin into a patient. The infusion of insulin may be controlled to occur at times and in amounts that are based, for example, on blood glucose measurements obtained from an embedded analyte sensor, such as a glucose sensor, in real-time.

There are two main types of blood glucose monitoring systems used by patients: single point or non-continuous and continuous. Non-continuous systems consist of meters and tests strips and require blood samples to be drawn from fingertips or alternate sites, such as forearms and legs. These systems rely on lancing and manipulation of the fingers or alternate blood draw sites, which can be extremely painful and inconvenient, particularly for children.

Continuous monitoring sensors are generally implanted subcutaneously and measure glucose levels in the interstitial fluid at various periods throughout the day, providing data that shows trends in glucose measurements over a short period of time. These sensors are painful during insertion and usually require the assistance of a health care professional. Further, these sensors are intended for use during only a short duration (e.g., monitoring for a matter of days to determine a blood sugar pattern). Subcutaneously implanted sensors may lead to infection and immune response complications. Another major drawback of currently available continuous monitoring devices is that they require frequent, often daily, calibration using blood glucose results that must be obtained from painful finger-sticks using traditional meters and test strips. This calibration, and recalibration, is required to maintain sensor accuracy and sensitivity, but it can be cumbersome as well as painful.

A typical glucose sensor works according to the following chemical reactions:
<MAT>
<MAT>.

In equation <NUM>, the glucose oxidase is used to catalyze the reaction between glucose and oxygen to yield gluconic acid and hydrogen peroxide (H<NUM>O<NUM>). The hydrogen peroxide reacts electrochemically as shown in Equation <NUM> and the resulting current can be measured by a potentiostat. These reactions, which occur in a variety of oxidoreductases known in the art, are used in a number of sensor designs.

In the case of three electrode design (working, counter and reference electrode), the reference electrode is not consumed and the as produced O<NUM> (Equation <NUM>) is reduced at the counter electrode as per reaction (<NUM>).

O<NUM> + <NUM><NUM>O + 4e- → 4OH+     (<NUM>).

In the case of two electrode design (working and reference electrode only), the following reaction occurs at the reference electrode.

Here it is worth discussing the relative merits of the <NUM>-electrode and <NUM>-electrode designs. In the case of <NUM>-electrode design, oxygen is consumed at the counter electrode which is also needed for the glucose oxidase (Equation <NUM>). However, the Ag/AgCl reference remains stable; albeit sensor dependence on oxygen increases. In the case of <NUM>-electrode design, sensor dependence of oxygen is lowered; albeit the AgCl is consumed over time (as per Equation <NUM>) and thus one must provide enough amount of AgCl adequate for lifetime of the sensor.

Planar flexible analyte sensor are known i. from <CIT>, <CIT>, <CIT>, <CIT>, and <CIT>.

As analyte sensor technology matures and new applications for sensor technology are developed, there is a need for improved analyte sensors, such as for continuous monitoring sensors for use over longer durations. Also, there is a need to develop advanced processes for sensor fabrication that will yield a factory calibrated analyte sensor without need for further external calibrations.

Further, there is a need for low cost, large scale production of analyte sensors, such as glucose sensors. Conventional batch processing is not amenable to high volume production or to significant cost reductions.

Accordingly, it is desirable to have an improved analyte sensor and related fabrication method that address the shortcomings of traditional sensor systems. Furthermore, other desirable features and characteristics will become apparent from the subsequent detailed description and the appended claims, taken in conjunction with the accompanying drawings and the foregoing technical field and background.

Analyte sensors and methods for fabricating analyte sensors are provided. In an exemplary embodiment, a planar flexible analyte sensor includes a flexible base layer and a first electrode formed from a layer of sputtered platinum on the base layer. Also, the analyte sensor includes an insulating dielectric layer over the base layer, wherein the insulating dielectric layer leaves a portion of the first electrode exposed. Further, the analyte sensor includes an electrochemical sensing stack over the exposed portion of the first electrode, including a glucose oxidase layer over the layer of sputtered platinum and a glucose limiting membrane over the glucose oxidase layer.

In another embodiment, an analyte sensor includes a polyester substrate, a layer of platinum on the polyester substrate, a protection layer over the layer of platinum, and an electrochemical sensing stack over the layer of platinum.

Another embodiment provides a planar flexible analyte sensor including a polyester base layer and a working electrode and a reference electrode formed from a layer of sputtered platinum on the base layer. Further, the planar flexible analyte sensor includes an insulating dielectric layer over the polyester base layer and layer of sputtered platinum. The insulating dielectric layer leaves a portion of the first electrode exposed and a portion of the second electrode exposed. Also, the planar flexible analyte sensor includes silver/silver chloride ink on the exposed portion of the reference electrode. The planar flexible analyte sensor further includes a glucose oxidase layer over the exposed portion of the working electrode and a glucose limiting membrane over the glucose oxidase layer.

In another exemplary embodiment, a method for fabricating a planar flexible analyte sensor includes sputtering platinum onto a polyester base layer to form a layer of platinum. The method includes patterning the layer of platinum to form working electrodes and additional electrodes. Further, the method includes forming an insulating dielectric layer over the base layer, wherein the insulating dielectric layer is formed with openings exposing portions of the working electrodes and portions of the additional electrodes. Also, the method includes partially singulating individual sensors from the base layer, wherein each individual sensor is connected to the base layer by a tab. The method further includes depositing an enzyme layer over the exposed portions of the working electrodes and coating the working electrodes with a glucose limiting membrane.

Another exemplary embodiment provides a method for fabricating a planar flexible analyte sensor. The method includes providing a polyester base layer with a first side sputtered with a layer of platinum and with a second side opposite the first side. The method includes patterning the layer of platinum to form working electrodes. Also, the method includes forming an insulating dielectric layer over the first side of the base layer, wherein the insulating dielectric layer is formed with openings exposing portions of the working electrodes. The method further includes printing silver/silver chloride ink over the second side of the base layer. The method also includes partially singulating individual sensors from the base layer, wherein each individual sensor is connected to the base layer by a tab. The method includes depositing an enzyme layer over the exposed portions of the working electrodes and coating the working electrodes with a glucose limiting membrane.

In another embodiment, a method for fabricating analyte sensors in a roll-to-roll process includes providing a roll of a polyester substrate having a first side coated with a layer of platinum. The method feeds the polyester substrate from the roll to an electrode patterning stage. Further, the method includes patterning the layer of platinum to form working electrodes and reference electrodes. The method feeds the polyester substrate to an insulation stage. The method includes forming an insulating dielectric layer over the polyester substrate. The method feeds the polyester substrate to an insulation curing stage. The method includes curing the insulating dielectric layer. The method feeds the polyester substrate to an ink printing stage. The method includes depositing silver/silver chloride ink over the reference electrodes. The method feeds the polyester substrate to a drying stage. The method includes drying the silver/silver chloride ink. The method feeds the polyester substrate to a punching stage. The method includes punching the polyester substrate to form ribbons, wherein each ribbon is connected to a remaining polyester substrate web by a tab, and wherein each sensor includes a working electrode and a reference electrode. The method feeds the remaining polyester substrate to an enzyme deposition stage. The method includes depositing an enzyme layer over the working electrodes. The method feeds the remaining polyester substrate to an enzyme curing stage. The method includes curing the enzyme layer. The method feeds the remaining polyester substrate to a membrane formation stage. The method includes coating the working electrodes with a glucose limiting membrane.

Also, while the preceding background discusses glucose sensing and exemplary analyte sensors are described as glucose sensors herein, such description is for convenience and is not limiting. The claimed subject matter may include any type of analyte sensor utilizing an embodiment of the sensor electrodes described herein.

Blood-glucose measurements may be employed in infusion systems for regulating a rate of fluid infusion into a body. In particular circumstances, a control system may be adapted to regulate a rate of insulin, glucagon, and/or glucose infusion into a body of a patient based, at least in part, on a glucose concentration measurement taken from a body (e.g., from an analyte sensor such as a glucose sensor).

According to certain embodiments, examples of analyte sensors as described herein may be implemented in a hospital environment to monitor levels of glucose in a patient. Alternatively, according to certain embodiments, examples of analyte sensors as described herein may be implemented in non-hospital environments to monitor levels of glucose in a patient. Here, a patient or other non-medical professional may be responsible for interacting with analyte sensors.

To maintain healthy glucose levels, a person with type <NUM> diabetes may manage their glycemia by monitoring blood glucose levels, controlling diet, exercise, and self-administering appropriate amounts of insulin at appropriate times. Deviations from such glycemic management, such as skipping an insulin bolus at meal time or underestimating the carbohydrate content of a meal may bring about prolonged hyperglycemia. Likewise, receiving too much insulin (e.g., by over-bolusing) for a given blood glucose level and/or meal may bring about severe hypoglycemia. Other external factors, such as exercise or stress, may also contribute to glycemic deviations.

Errors in reading glucose levels may contribute to providing too much or too little insulin. Therefore, sensor accuracy is of utmost concern. Further, sensor accuracy must be maintained during the lifetime of a continuous glucose monitoring device. There is desire for longer life continuous glucose monitoring devices, i.e., continuous glucose monitoring devices that are implanted for a longer duration, such as for seven to fourteen days, or longer. Therefore, in the future, sensor accuracy must be maintained in vivo for seven to fourteen days, or longer. Continuous glucose monitoring sensors provide the ability to continuously track glucose levels in a patient, and to correlate them to his or her physical activity and diet, thereby providing for therapy decisions as well as for adjustments, as necessary.

By more accurately monitoring a patient's glucose level and maintaining appropriate infusion rates, extreme glycemic variations may be reduced or avoided altogether. This may provide a patient with improved glycemic control in circumstances in which they would otherwise be exposed to undesirable extremes of glycemia.

Embodiments herein provide for improved accuracy as compared to current commercialized sensors, and will reduce sensor cost through new manufacturing processes. For example, sensors herein may utilize excimer-patterned electrodes formed from platinum-sputtered layers on polyester film substrates.

<FIG> is a cross sectional view of an example analyte sensor <NUM> for use with a glucose control system in accordance with an embodiment. Particular embodiments of the analyte sensor <NUM> include a planar flexible polymeric base layer <NUM>, such as a polyester film or substrate. As shown, the base layer <NUM> includes a first side <NUM> and an opposite second side <NUM> and an end <NUM>. An exemplary base layer <NUM> has a thickness of from about <NUM> micrometers (µm) (about <NUM> mil) to about <NUM> (about <NUM> mil), such as about <NUM> (about <NUM> mil) or about <NUM> (about <NUM> mil).

In certain embodiments, the base layer <NUM> has a surface roughness of from about <NUM> to about <NUM>, for example, from <NUM> to about <NUM>, such as from about <NUM> to <NUM>, or from about <NUM> to <NUM>, or from about <NUM> to <NUM>, or from about <NUM> to <NUM>, or from about <NUM> to <NUM>.

Further, a platinum layer <NUM> is formed on the first side <NUM> of the base layer <NUM>. In exemplary embodiments, the platinum layer <NUM> is formed by sputtering platinum onto the first side <NUM> of the base layer <NUM>. An exemplary platinum layer <NUM> may have a thickness of from about <NUM> to about <NUM>, such as from about <NUM> to about <NUM>.

During in vivo use, platinum is challenged by the immune response that the body elicits upon sensor implantation. For example, the constant mechanical forces exerted by the body tissue around the implant may compromise the adhesion of platinum to polyester substrates. Therefore, the first side <NUM> of the base layer <NUM> may undergo surface modification to modulate adhesion between the platinum layer <NUM> and the base layer <NUM>. Such surface modification may allow for controlled and reproducible tuning of electrode surface area to improve in vivo sensor performance. Also, surface modification may provide for grafting functional moieties to the first side <NUM> of the base layer <NUM> to increase adhesion. Surface modification may be performed by plasma pretreatment of the base layer <NUM>. Other processes may be performed to roughen the surface of the base layer <NUM>.

Also, the platinum layer <NUM> may be formed with an exterior surface <NUM> having a selected surface roughness. Sputtering of platinum on polyester substrates is a well-known process that yields high uniformity and reproducibility in the surface roughness of the exterior surface <NUM>. Surface roughness of the exterior surface <NUM> of platinum layer <NUM> is a controlling factor for electrode surface area. Through adjusting the process parameters of the platinum sputtering process, the surface roughness of the exterior surface <NUM> of platinum layer <NUM>, and electrodes, may be controlled.

As shown, the analyte sensor <NUM> is formed with electrodes necessary for sensor operation. For example, the analyte sensor <NUM> may be formed with working, reference, and counter electrodes. In <FIG>, two electrodes <NUM> and <NUM> are illustrated for simplicity; however, the analyte sensor <NUM> may include one or more working, reference, and counter electrodes. In certain embodiments, the analyte sensor <NUM> may include pairs of working electrodes and counter electrodes.

The electrodes <NUM> and <NUM> may be formed by patterning the platinum layer <NUM>. For example, a laser ablation process may be performed to pattern the platinum layer <NUM>. In an exemplary embodiment, an excimer laser, such as a <NUM> nanometer excimer laser, is utilized to pattern the platinum layer <NUM> with ultraviolet light to form the electrodes <NUM> and <NUM>. Laser ablation provides for high throughput during sensor fabrication processing and is highly reproducibly. For example, in certain embodiments, electrodes for eighteen analyte sensors <NUM> can be patterned per second by the laser ablation process.

As shown, the analyte sensor <NUM> further includes an insulating dielectric layer <NUM> over the first side <NUM> of the base layer <NUM>. An exemplary insulating dielectric layer <NUM> may be a polymer that is crosslinked by ultra violet radiation or by a thermal process such that after crosslinking the insulation dielectric is impermeable to solvents and water and other electrochemically active constituents in the analyte containing fluid. The insulating dielectric layer <NUM> is provided to prevent diffusion of the electrochemically active constituent to the electrode's electrochemically active surface for the purpose of accurately controlling the electrode signal level.

The electrode signal is proportional to the surface area that is exposed to the analyte containing fluid. An insulating dielectric layer <NUM> that is impermeable to electrochemically active constituents in the analyte containing fluid can be applied and patterned to define the electrode's electrochemically active surface area. Methods of applying and patterning the dielectric layer include screen printing, digital drop on demand, transfer pad printing, gravure coating or other photolithography patterned coating method known to those skilled in the art.

An exemplary insulating dielectric layer <NUM> can be an acrylic polymer that is thermally crosslinked. An exemplary insulating dielectric layer <NUM> has a thickness of from about <NUM> to about <NUM>, such as about <NUM>.

In exemplary embodiments, the insulating dielectric layer <NUM> is formed by screen printing or rotary printing the insulating dielectric material. In certain embodiments, the insulating dielectric layer <NUM> is patterned or otherwise formed with openings <NUM> that leave a portion <NUM> of electrode <NUM> exposed and a portion <NUM> of electrode <NUM> exposed. In other words, after formation of the insulating dielectric layer <NUM>, the portion <NUM> of the electrode <NUM> and the portion <NUM> of the electrode <NUM> are not covered by the insulating dielectric layer <NUM>. The openings <NUM> define the geometric surface area of the electrodes <NUM> and <NUM>. In other words, the exposed portions <NUM> and <NUM> of the electrodes <NUM> and <NUM> are the geometric surface areas of the electrodes <NUM> and <NUM>.

In exemplary embodiment, electrode <NUM> is a working electrode, electrode <NUM> is a counter electrode, and the width of the insulating dielectric layer <NUM> is modulated so that the end-to-end distance <NUM> between the openings <NUM> of platinum working electrode <NUM> and platinum counter electrode <NUM> is changed to improve overall sensor sensitivity and to lower sensor dependence on oxygen concentration. The end-to-end distance between the openings can be from about <NUM> microns to about <NUM> microns, such as from about <NUM> microns to about <NUM> microns, for example about <NUM> microns.

Thus, <FIG> may be considered to illustrate a single pair of working and counter electrodes <NUM> and <NUM> with a selected intrapair distance <NUM>. It is contemplated that the sensor <NUM> include multiple working electrodes (WE) and counter electrodes (CE), wherein each WE/CE pair has a specific, independently selected end-to-end distance <NUM> between the openings <NUM> of the working and counter electrodes. In other words, the sensor <NUM> may be provided with a plurality of WE/CE pairs, each having an independently selected intrapair distance. While WE/CE pairs may be provided with different intrapair distances from one another, certain WE/CE pairs may have a same intrapair distance.

The differential response from these multiple WE/CE pairs can provide insights on sensor dependence on oxygen which can used to calibrate the sensor performance against sensitivity drifts. For example, sensitivity can drift due to sensor degradation caused by exposure to various electroanalytical species such as oxygen radicals. In addition, post-implantation effects such as biofouling and foreign body response also contribute to passivation of the electro catalytic activity of the electrode.

In <FIG>, further processing may be performed on the analyte sensor <NUM>. In <FIG>, electrode <NUM> is processed to form a working electrode and electrode <NUM> is processed to form a reference electrode (as noted above, multiple electrodes are not illustrated simply for purposes of clarity). For a reference electrode <NUM>, a silver/silver chloride (Ag/AgCl) ink layer <NUM> is formed over the electrode <NUM>. The silver/silver chloride layer <NUM> may be selectively deposited by screen printing or rotary printing. Unlike conventional electro-deposition or electro-oxidation processes, silver chloride loading is not limited by the surface area of the electrode when screen printing or rotary printing silver/silver chloride ink. In an exemplary embodiment, the silver/silver chloride layer <NUM> is overloaded such that the amount of AgCl is always in excess as compared to what is needed for sensor operation over <NUM> days. In some cases the ratio of AgCl to Ag is greater than <NUM> and can be from about <NUM> to about <NUM>, such as about <NUM>. It is noted that the silver/silver chloride (Ag/AgCl) ink layer <NUM> may be formed before or after formation of the insulating dielectric layer <NUM> but a preferred embodiment includes depositing the silver/silver chloride ink before the dielectric material.

In an exemplary embodiment, the silver/silver chloride ink has a formulation of micro and nano particles of silver and silver chloride in a polymeric binder to enhance overloading of the silver/silver chloride ink during screen printing or rotary printing. In an exemplary embodiment, the silver/silver chloride layer has a thickness (or height) of from about <NUM> to about <NUM>, such as about <NUM>. In an exemplary embodiment, the reference electrode is made up of metal oxide micro and nanoparticles loaded within a polymeric binder. An exemplary metal oxide is Iridium Oxide.

As noted above, during in vivo use, platinum is challenged by the immune response that the body elicits upon sensor implantation. For example, degradation of the platinum may be induced by biofouling and the body's immune response. Unless provided for, the platinum may not withstand the degradation induced by biofouling and the body's immune response. Therefore, the analyte sensor <NUM> may be provided with a protection layer <NUM> over the first side <NUM> of the base layer <NUM> for protecting the platinum from biofouling and other immune response degradation. An exemplary protection layer <NUM> may be a hydrophilic hydrogel layer and can be made from a wide variety of materials known to be suitable for such purposes, e.g., polyvinyl alcohol, poly (N-isopropylacrylamide), poly (N-vinylpyrrolidone), polyethylene glycol, polyurethane, poly acrylic acid, cellulose acetates, Nafion, polyester sulfonic acids hydrogels or any other suitable hydrophilic membranes known to those skilled in the art.

An exemplary protection layer <NUM> has a thickness of from about <NUM> to about <NUM>, such as about <NUM>. As shown in <FIG>, the protection layer <NUM> covers the entire top side <NUM> of the base layer <NUM> and completely encapsulates the platinum layers of the electrodes <NUM> and <NUM>.

In exemplary embodiments, the protection layer <NUM> is formed by screen printing, rotary printing, spray coating, dip coating, spin coating or brush coating. In certain embodiments, the diffusion properties of the protection layer <NUM> are carefully controlled so that the magnitude of the sensor signal is not compromised due to smaller diffusion coefficients that will not let hydrogen peroxide pass through, leading to minimal or no signal as per Equation <NUM> above. In such embodiment, hydrophobic moieties such as acrylate polymers or surfactants or oxygen loading species (such as fluorocarbon compounds) or oxygen bearing enzymes such as myoglobin or hemoglobin or oxygen generating enzymes such as catalase are incorporated within the hydrogel layer to improve adhesion and tune permeability to hydrogen peroxide and oxygen.

<FIG> illustrates the silver/silver chloride layer <NUM> formed before the protection layer <NUM> such that the silver/silver chloride layer <NUM> is disposed under the protection layer <NUM>. However, it is noted that the order of formation may be reversed, such that the protection layer <NUM> lies under the silver/silver chloride layer <NUM>.

In <FIG>, the analyte sensor <NUM> further includes an electrochemical sensing stack <NUM> over the exposed portion <NUM> of the working electrode <NUM>. The electrochemical sensing stack <NUM> may include a plurality of layers that are not individually illustrated in <FIG>. In an exemplary embodiment, the electrochemical sensing stack <NUM> includes an analyte sensing layer, such as an enzyme layer, for example a glucose oxidase layer. An exemplary glucose oxidase layer has an activity of from about <NUM> KU/mL to about <NUM> KU/mL, such as from about <NUM> to about <NUM> KU/mL, for example about <NUM> KU/mL. Further, an exemplary glucose oxidase layer has a thickness of from about <NUM> to about <NUM> micrometers (µm), such as from about <NUM> to about <NUM>, for example from about <NUM> to about <NUM>, such as about <NUM>. In an exemplary embodiment, the enzyme layer is deposited over the working electrode <NUM> by rotary screen printing. In another embodiment, the enzyme layer is deposited over the working electrode <NUM> by an aerosol-based drop on demand technology. In another embodiment, the enzyme layer is deposited over the working electrode <NUM> by spin coating or spray coating. In another embodiment, the enzyme layer is deposited over the working electrode <NUM> by one of the above mentioned techniques and further crosslinked by ultra violet radiation or exposure to vapors of crosslinking agents such as glutaraldehyde.

In certain embodiments, the electrochemical sensing stack <NUM> may include additional layers, such as a protein layer. Typically, a protein layer includes a protein such as human serum albumin, bovine serum albumin or the like.

In certain embodiments, the electrochemical sensing stack <NUM> may include an adhesion promoter layer disposed over the analyte sensing or enzyme layer in order to facilitate contact and/or adhesion between the analyte sensing layer and another overlying layer. The adhesion promoter layer can be made from any one of a wide variety of materials known in the art to facilitate the bonding between such layers. Typically, the adhesion promoter layer includes a silane compound. In alternative embodiments, protein or like molecules in the analyte sensing layer can be sufficiently crosslinked or otherwise prepared to allow the analyte modulating membrane layer to be disposed in direct contact with the analyte sensing layer in the absence of an adhesion promoter layer. In certain embodiments, additional layers such as an interference rejection layer may be included in the electrochemical sensing stack <NUM>. Such layers may be formed by rotary or screen printing or spin coating or spray coating or through chemical vapor deposition. In another embodiment, the adhesion promoter layer is deposited over the working electrode <NUM> by one of the above mentioned techniques and further crosslinked by ultra violet radiation or exposure to vapors of crosslinking agents such as glutaraldehyde.

Thus, in certain embodiments, the stack <NUM> includes a protein layer, such as a human serum albumin (HSA) layer, overlying the glucose oxidase layer, and an adhesion promoting layer over the protein layer. In other embodiments, no protein layer or adhesion promoting layer are included in the electrochemical sensing stack <NUM>.

In <FIG>, the electrochemical sensing stack <NUM> is shown to further include an analyte modulating layer <NUM>, such as a glucose limiting membrane (GLM) <NUM>, over the enzyme layer. The analyte modulating layer <NUM> is provided to regulate analyte contact with the analyte sensing layer or enzyme layer. For example, the analyte modulating membrane layer can be a glucose limiting membrane, which regulates the amount of glucose that contacts an enzyme such as glucose oxidase that is present in the analyte sensing layer. Such glucose limiting membranes can be made from a wide variety of materials known to be suitable for such purposes, e.g., silicone compounds such as polydimethyl siloxanes, polyurethanes, polyurea cellulose acetates, Nafion, polyester sulfonic acids (e.g. Kodak AQ), hydrogels or any other suitable membranes known to those skilled in the art. In an exemplary embodiment, the thickness of the glucose limiting membrane <NUM> is from about <NUM> to about <NUM> micrometers (µm), such as from about <NUM> to about <NUM>, for example from about <NUM> to about <NUM>, such as about <NUM>.

As shown, the glucose limiting membrane <NUM> may be formed around the entire end <NUM> of the analyte sensor <NUM>, i.e., over both the first side <NUM> and the second side <NUM> of the base layer <NUM>, including over reference electrodes and counter electrodes. In an exemplary embodiment, the glucose limiting membrane <NUM> may be formed by dip coating the end <NUM> of the analyte sensor <NUM>, such that the glucose limiting membrane <NUM> encapsulates the analyte sensor <NUM> for insertion into a patient's interstitial fluid.

In addition to the glucose limiting membrane <NUM>, other membranes may be formed over the end <NUM> of the analyte sensor <NUM> to suppress the foreign body response upon implantation. For example, a foreign body response (FBR) membrane may be formed over the glucose limiting membrane <NUM>, such as by dip coating the end <NUM> of the analyte sensor <NUM>.

<FIG> illustrates an alternate or additional embodiment of analyte sensor <NUM>. In <FIG>, the reference electrode <NUM> is not formed from the platinum layer <NUM> on the first side <NUM> of the base layer <NUM>. Rather, the reference electrode <NUM> is formed on the second side <NUM> of the base layer <NUM>. Specifically, the reference electrode <NUM> is formed by a silver/silver chloride ink layer <NUM> deposited over the second side <NUM> of the base layer <NUM>. The silver/silver chloride ink may be selectively deposited onto the second side <NUM> of the base layer <NUM> by screen printing or rotary printing. In such embodiments, the entire second side <NUM> could be overprinted or overloaded with silver/silver chloride ink to allow more loading of silver chloride. Further, such an embodiment eliminates positional registration process capability to reduce overall fabrication process error. As shown in <FIG>, the glucose limiting membrane <NUM> completely covers the reference electrode <NUM> formed by the silver/silver chloride ink layer <NUM> on the second side <NUM> of the base layer <NUM>.

Referring now to <FIG>, a system <NUM> and method for fabricating analyte sensors <NUM> is described. As shown, the system <NUM> fabricates analyte sensors in a roll-to-roll process by processing a roll <NUM> of a flexible substrate, such a polyester film, that includes a layer of sputtered platinum as described above. In the system <NUM>, the polyester substrate <NUM> is fed from the roll <NUM> to an electrode patterning stage <NUM>. At the electrode patterning stage <NUM>, the layer of platinum is patterned, such as by laser ablation, to form working electrodes and counter electrodes, and optionally reference electrodes, depending on the desired analyte sensors. Then, the polyester substrate <NUM> is fed from the electrode patterning stage <NUM> to an insulation stage <NUM>. At the insulation stage <NUM>, an insulating dielectric material is selectively deposited over the polyester substrate <NUM>. For example, an insulating dielectric material may be screen printed or rotary printed over the polyester substrate.

The polyester substrate <NUM> is then fed from the insulation stage <NUM> to an insulation curing stage <NUM> where the insulating dielectric material is cured to form the insulating dielectric layer. For example, an ultraviolet light curing process may be performed. Then, the polyester substrate <NUM> is fed from the insulation curing stage <NUM> to an ink printing stage <NUM>. At the ink printing stage <NUM>, silver/silver chloride ink is deposited over the reference electrodes, if such reference electrodes have been formed from the sputtered platinum. Alternatively, the silver/silver chloride ink is deposited over the second side or backside of the polyester substrate to form the reference electrodes. The silver/silver chloride ink may be selectively deposited by screen printing or by rotary printing.

As shown, the polyester substrate <NUM> is then fed from the ink printing stage <NUM> to a drying stage <NUM>. At the drying stage <NUM>, the silver/silver chloride ink is dried to form the silver/silver chloride ink layer on the polyester substrate. After drying, the polyester substrate <NUM> is then fed to a protection layer stage <NUM> where a protection material is deposited over the polyester substrate <NUM>. Specifically, the protection material is deposited over the entire side of the polyester substrate <NUM> on which the sputtered platinum electrodes are located. The protection material may be deposited by screen printing or by rotary printing. The polyester substrate <NUM> is then fed to the curing stage <NUM> where the protection material is cured to form a protection layer.

The polyester substrate <NUM> is then fed to a punching stage <NUM>. At the punching stage <NUM>, the polyester substrate is punched, or otherwise cut, to form partially singulated ribbons for forming singulated analyte sensors. For example, the polyester substrate may be laser cut to form the partially singulated ribbons. As shown, the polyester substrate <NUM> may be passed under a vision alignment device <NUM> to ensure that the polyester substrate <NUM> is properly aligned for punching. After punching, each ribbon is still connected to a remaining polyester substrate web by a tab. In exemplary embodiments, each ribbon or analyte sensor includes a working electrode and a reference electrode. Specifically, each ribbon or analyte sensor includes the desired number of working electrodes, reference electrodes and counter electrodes without further electrode formation.

A punched out or scrap portion <NUM> of the polyester substrate <NUM> may be wound up into a roll <NUM> from the punching stage <NUM>. The remaining polyester substrate <NUM> (including the singulated analyte sensors in the form of ribbons) is fed to an enzyme deposition stage <NUM>. At the enzyme deposition stage <NUM>, an enzyme, such as glucose oxidase, is deposited over the working electrodes. For example, the enzyme may be screen printed or rotary printed onto the polyester substrate. Alternatively, the enzyme may be deposited by an aerosol-based drop on demand technology. In addition to the enzyme, other materials for forming an electrochemical sensing stack may be deposited over the polyester substrate at the enzyme deposition stage <NUM>.

Then, the polyester substrate <NUM> is fed to an enzyme curing stage <NUM>. At the enzyme curing stage <NUM>, the enzyme (and other deposited materials) is cured, such as by an ultraviolet light photo-initiated curing process. During curing, the enzyme crosslinks and becomes immobilized. The polyester substrate <NUM> may then be fed to a membrane formation stage <NUM>. At the membrane formation stage <NUM>, the end of each ribbon may be coated with a glucose limiting membrane and other desired membranes. For example, the end of each ribbon may be dip coated or slot coated to form the desired membranes.

In certain embodiments, the system <NUM> may terminate the roll-to-roll processing at this stage and produce a polyester substrate <NUM> including individual analyte sensors, in the form of ribbons connected to the polyester substrate web, ready for further integration with production of a glucose monitoring system. Alternatively, roll-to-roll processing may continue with processing the individual analyte sensors on polyester substrate <NUM>. Specifically, the individual analyte sensors on polyester substrate <NUM> may be processed through a functionality check stage <NUM>. At the functionality check stage <NUM>, each analyte sensor is exposed to a buffer solution containing glucose, and the sensor's signal-time profile is recorded and evaluated.

Further, the polyester substrate <NUM> may be removed from the functionality check stage <NUM> and may be fed to a singulation stage <NUM>. At the singulation stage <NUM>, the individual analyte sensors, in the form of ribbons connected to the polyester substrate web, may be singulated through complete separation from the polyester substrate web. The singulation stage <NUM> may be part of a final system assembly and packaging system separate from the roll-to-roll processing system <NUM> described here.

It is noted that the system <NUM> of <FIG> may include additional elements, such as idler wheels, driven wheels, vacuum boxes, and additional, alternative, or repetitive process stages, for the fabrication of the analyte sensors.

<FIG> is a top view of a partially singulated substrate <NUM>, as fabricated according to the method of <FIG>. Specifically, the partially singulated substrate <NUM> is illustrated as formed after being punched out, after enzyme layer formation, and after membrane formation, such as after membrane formation stage <NUM> or after functionality check stage <NUM> of <FIG>. Further, the partially singulated substrate <NUM> is illustrated as before complete separation of analyte sensors from the polyester substrate at singulation stage <NUM> of <FIG>.

As shown in <FIG>, the partially singulated substrate <NUM> includes individual analyte sensors <NUM>, in the form of ribbons. Each individual analyte sensor <NUM> is connected to the polyester substrate web <NUM> by a tab at end <NUM>. Complete singulation and separation of the individual analyte sensors <NUM> may occur by severing at each tab at end <NUM>. As shown, each analyte sensor <NUM> terminates at an end <NUM>. End <NUM> may be dipped or otherwise coated with the membrane-forming materials at membrane formation stage <NUM> of <FIG>. The gaps <NUM> between adjacent analyte sensors <NUM> are formed during the punching out process by removal of portions of the polyester substrate, such as the punched out or scrap portion <NUM> in <FIG>.

<FIG> illustrates a system <NUM> and method for fabricating the platinum-sputtered polyester substrate that is introduced in <FIG> on roll <NUM>. In <FIG>, a polyester substrate <NUM> is provided. The polyester substrate <NUM> may be introduced to a polyester surface modification stage <NUM>. At the polyester surface modification stage <NUM>, the surface of a side or both sides of the polyester substrate <NUM> is modified. Specifically, the polyester surface may be roughened to improve adhesion between the polyester and the platinum layer to be formed thereon. Also, polyester surface modification may provide for grafting functional moieties to the surface to increase adhesion. Surface modification may be performed by plasma pretreatment of the polyester substrate <NUM>. Other processes for surface modification may include electric discharge, surface grafting, flame treatment, UV irradiation or wet chemical etching.

Other processes may be performed to roughen the surface of the polyester substrate <NUM> and include wet chemical etching, hydrolysis in a basic solution, acid washing or laser irradiation.

As shown, the polyester substrate <NUM> is then fed to a platinum deposition stage <NUM>. At the platinum deposition stage <NUM>, platinum is deposited onto a side of the polyester substrate <NUM>. For example, platinum may be sputtered onto the polyester substrate. Sputtering is a physical vapor deposition method. Sputtering can be carried out using commercially available sputtering reactors using an RF (radio frequency). Magnetron sputtering can also be used. Magnetron sputtering uses a magnetic field to concentrate electrons near the target surface to increase the deposition rate. The thickness of an exemplary sputtered platinum layer is from about <NUM> to about <NUM>, for example, from about <NUM> to about <NUM>. When multiple layers are deposited, the total thickness of the layers may have a thickness within the foregoing ranges.

The characteristics of the thin film of platinum that is produced through sputtering vary based on the process parameters, such as characteristics of the platinum sputtering target material, including purity and microstructure, sputtering rate, the energy with which the sputtered platinum atoms reach the polyester substrate, temperature, and/or other parameters.

Control of the process parameters may produce a platinum-sputtered polyester substrate <NUM> having desired properties. For example, through adjusting the process parameters of the platinum sputtering process, the surface roughness of the exterior surface of the platinum layer may be controlled. Alternatively, the platinum-sputtered polyester substrate <NUM> may undergo further processing. For example, the platinum-sputtered polyester substrate <NUM> may be introduced to a platinum surface modification stage <NUM>. At the platinum surface modification stage <NUM>, a plasma surface modification process or other surface modification process may be performed to adjust the surface characteristics of the platinum layer as desired. Thereafter, the platinum-sputtered polyester layer <NUM> may be wound on a roll <NUM> for use in the fabrication method <NUM> of <FIG>.

It should be noted that although aspects of the above methods, systems and sensors have been described in particular orders and in particular arrangements, such specific orders and arrangements are merely examples and claimed subject matter is not limited to the orders and arrangements as described.

Claim 1:
A planar flexible analyte sensor (<NUM>) comprising:
a polyester base layer (<NUM>) having a layer of sputtered platinum formed directly on the polyester base layer (<NUM>);
a working electrode (<NUM>) and a reference electrode (<NUM>) formed by patterning the layer of sputtered platinum on the base layer by laser ablation;
an insulating dielectric layer (<NUM>) over the polyester base layer and the layer of sputtered platinum, wherein the insulating dielectric layer leaves a portion (<NUM>) of the working electrode exposed and a portion (<NUM>) of the reference electrode exposed;
silver/silver chloride ink (<NUM>) on the exposed portion of the reference electrode;
a glucose oxidase layer over the exposed portion of the working electrode; and
a glucose limiting membrane over the glucose oxidase layer.