Methods, sensors, and wearable devices for rapid detection of myocardial infarction

Abstract

The present disclosure provides devices, systems incorporating the devices, methods for manufacturing the devices, and methods of using the devices for carrying out assays, such as chemical and / or biological assays. The systems include a wearable device including a biosensor having a plurality of electrodes. A functionalized coating is coupled to an electrode of the biosensor. A controller may sense an electrochemical response of the biosensor, and determine whether a health-related biomarker is present based on the electrochemical response. The functionalized coating may include a sulfonated block copolymer such as sulfonated styrene-isobutylene-styrene (SIBS). The health-related biomarker may include cardiac troponin I (cTnI).

Description

Docket No. 23-002-UPR / 71900-431712METHODS, SENSORS, AND WEARABLE DEVICES FOR RAPID DETECTION OF MYOCARDIAL INFARCTIONCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 USC § 119(e) of U.S. Provisional Application Serial No. 63 / 724,613, filed on November 24, 2024, the entire disclosure of which is incorporated herein by reference.TECHNICAL FIELD

[0002] The present disclosure relates to devices and methods for detection of health- related biomarkers. More particularly, the disclosure relates to methods and devices for detection of myocardial infarction using biosensors.BACKGROUND

[0003] In the United States, heart disease is the leading cause of death. Further, approximately 800,000 myocardial infarctions (heart attacks) occur every year. Early detection of myocardial infarction is crucial for patient survival, as late treatment can cause irreversible damage or complications that can lead to death.

[0004] Traditional electrocardiograms (ECG) alone cannot provide specific detection of a myocardial infarction. Cardiac troponin T (cTnl) is a regulatory protein unique to the myocardium that controls the calcium-mediated interaction between actin and myosin. This cardiac troponin can be detected after a myocardial infarction in the patient’s serum and saliva. Current cardiac troponin tests are invasive (requiring a blood sample), have complex preparation procedures, present high toxicity, and / or are expensive.BRIEF SUMMARY

[0005] The present disclosure provides devices, systems, and methods for detection of health-related biomarkers. In some embodiments, a wearable device includes a biosensor, a functionalized coating, a controller coupled to the biosensor, and a wearable feature coupled to the biosensor. The biosensor includes a plurality of electrodes, and the functionalized coating is coupled to a first electrode of the plurality of electrodes. The functionalized coating comprises a sulfonated block copolymer. The controller is configured to sense an electrochemical response of the biosensor, and to determine whether a health-related biomarker is present based on the electrochemical response.Docket No. 23-002-UPR / 71900-431712

[0006] In some embodiments, the sulfonated block copolymer comprises a sulfonated styrene -isobutylene-styrene (SIBS) block copolymer. In some embodiments, the health-related biomarker comprises cardiac troponin I (cTnl).

[0007] In some embodiments, to sense the electrochemical response of the biosensor may include to apply electrochemical impedance spectroscopy, cyclic voltammetry, linear sweep voltammetry, or differential pulse voltammetry to the biosensor.

[0008] In some embodiments, the plurality of electrodes may include a screen printed electrode. In some embodiments, the plurality of electrodes may include gold or silver. In some embodiments, the plurality of electrodes may include a conductive carbon-based polymer. In some embodiments, the plurality of electrodes may include laser-scribed electrodes.

[0009] In some embodiments, the wearable feature may include a strap, an adhesive patch, a glove, an article of clothing, or an implant.

[0010] According to another aspect, a method for manufacturing a biosensor includes providing a film substrate; scribing a plurality of electrodes on a surface of the film substrate with a laser, wherein the laser is configured with first manufacturing parameters; mounting the film substrate to a base; and positioning a cover on the film substrate after scribing the plurality of electrodes, wherein the cover comprises an aperture that when positioned on the film substrate exposes at least part of each of the plurality of electrodes.

[0011] In some embodiments, the first manufacturing parameters may include laser power, speed, laser focus, and / or laser dot pitch.

[0012] In some embodiments, the film substrate comprises a non-conductive polymer film substrate. In some embodiments, the film substrate comprises a polyimide (PI) film substrate.

[0013] In some embodiments, the base comprises a poly(methyl methacrylate) (PMMA) base. In some embodiments, the cover comprises a semi-rigid polymeric material. In some embodiments, the cover comprises a vinyl cover.

[0014] In some embodiments, the method may further include depositing a SIBS block copolymer on a first electrode of the plurality of electrodes.

[0015] According to another aspect, a method for manufacturing a biosensor includes training, by a computing device, a generator model and a discriminator model with training data, the training data comprising electrical parameters for laser-scribed electrodes associated with manufacturing parameters; generating, by the computing device, a plurality of candidate manufacturing parameters with the generator model in response to training the generator model; selecting, by the computing device, a first candidate manufacturing parameter of the candidateDocket No. 23-002-UPR / 71900-431712 manufacturing parameters with the discriminator model in response to training the discriminator model; and manufacturing a test biosensor with the first candidate manufacturing parameter.

[0016] In some embodiments, the method may further include evaluating one or more electrical parameters of the test biosensor; and updating, by the computing device, the generator model and the discriminator model with the first candidate manufacturing parameter and the electrical parameters of the test biosensor.

[0017] In some embodiments, the manufacturing parameters may include laser power, speed, laser focus, and laser dot pitch.

[0018] The foregoing has outlined rather broadly the features and technical advantages of the present disclosure in order that the detailed description that follows may be better understood. Additional features and advantages of the disclosure will be described hereinafter that form the subject of the claims of this application. It should be appreciated by those skilled in the art that the conception and the specific embodiments disclosed may be readily utilized as a basis for modifying or designing other embodiments for carrying out the same purposes of the present disclosure. It should also be realized by those skilled in the art that such equivalent embodiments do not depart from the spirit and scope of the disclosure as set forth in the appended claims.BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0019] A detailed description of the invention is hereafter described with specific reference being made to the drawings in which:

[0020] FIG. 1 is a perspective view of a biosensor for detection of a health-related biomarker;

[0021] FIG. 2 is an exploded view of the biosensor of FIG. 1 :

[0022] FIG. 3 is a schematic view of a wearable device including a biosensor for detection of a health-related biomarker;

[0023] FIG. 4 is a simplified flow diagram of at least one method for detection of a health- related biomarker using a biosensor of FIGS. 1-3;

[0024] FIG. 5 is a chart illustrating experimental results that may be achieved with a biosensor of FIGS. 1-4;

[0025] FIG. 6 is a simplified flow diagram of at least one embodiment of a method for manufacturing a biosensor of FIGS. 1-4;

[0026] FIG. 7 is a simplified diagram of a reaction mechanism of a sulfonated block copolymer that may be used with the method of FIG. 6.Docket No. 23-002-UPR / 71900-431712

[0027] FIG. 8 is a simplified flow diagram of at least one embodiment of a method for manufacturing a laser-scribed electrode that may be used with a biosensor of FIGS. 1-4; and

[0028] FIG. 9 is a simplified flow diagram of at least one embodiment of a method for optimizing manufacturing parameters for a laser-scribed electrode manufactured according to the method of FIG. 8.DETAILED DESCRIPTION

[0029] Various embodiments are described below with reference to the drawings in which like elements generally are referred to by like numerals. The relationship and functioning of the various elements of the embodiments may better be understood by reference to the following detailed description. However, embodiments are not limited to those illustrated in the drawings or explicitly described below.

[0030] Referring now to FIGS. 1 and 2, an embodiment of a biosensor 100 of the present disclosure is depicted. The illustrative biosensor 100 includes multiple electrodes 102. Illustratively, the electrodes 102 include a counter electrode 104, a working electrode 106, and a reference electrode 108. Each electrode may be embodied as a metallic material, a carbon-based material, or another conductive material. For example, in some embodiments, the biosensor 100 may include commercially available carbon-based and / or gold electrodes 102. In an embodiment, the biomarker 100 uses a screen-printed electrode (SPE) that is commercially available from BVT Technologies, Strazek, Czech Republic. In this illustrative embodiment, the SPE has silver as a reference electrode, graphite as a working electrode, and a counter electrode. Additionally or alternatively, in some embodiments the biosensor 100 may include carbon or gold working electrodes. Gold electrodes may offer an order of magnitude greater electrical signal compared to carbon-based electrodes. However, carbon-based electrodes may be more sensitive when measuring electrolytes than gold electrodes, while gold electrodes may be more sensitive when measuring cTnl, and possibly proteins, in a more general sense. Commercially available SPEs may provide suitable characteristics, such as low cost, disposability, ease of use, and portability.

[0031] In the illustrative embodiment, the electrodes 102 are laser-scribed electrodes (LSEs). As described further below, the electrodes 102 of the LSEs may be formed from a graphene- or graphite-like carbon material that is formed on the surface of the biosensor using a laser, such as a laser cutter or laser engraver. Potential embodiments of methods for manufacturing laser-scribed electrodes 102 are described further below in connection with FIGS. 8-9. Additionally or alternatively, the electrodes 102 may be embodied as screen-printed electrodes (SPE), which may be commercially available. For example, as described above, theDocket No. 23-002-UPR / 71900-431712 biosensor 100 may include an SPE that is commercially available from BVT Technologies of the Czech Republic.

[0032] One or more of the electrodes 102 is coated with a functionalized coating 110 including a sulfonated block copolymer. For example, in the illustrative embodiment, the working electrode 106 is coated with the functionalized coating 110. As described further below, the functionalized coating 110 interacts with a particular biomarker, and an electrochemical response of the electrodes 102 thus may be used to detect the biomarker. In the illustrative embodiment, the sulfonated block copolymer of the functionalized coating 110 is sulfonated styrene-isobutylene-styrene (SIBS) block copolymer. The nano-scale structure of SIBS interacts with cardiac troponin I (cTnl), which is a biomarker related to myocardial infarction. Additionally or alternatively, in some embodiments the sulfonated block copolymer may be embodied as sulfonated poly(arylene ether sulfone) (SPAES), sulfonated poly(arylene ether ketone) (SPAEK), and / or sulfonated poly(arylene ether ketone sulfone) (SPAKES). In some embodiments, those sulfonated block copolymer may have increased elastomeric domains with improved membrane properties, while maintaining high conductivity, stability, and other parameters.

[0033] Block copolymers (BCPs) are a type of polymer with potential adsorption sites for biomarkers. Some advantages of BCPs include the possible functionalization of specific functional groups, that may improve the physical and chemical properties of the polymer. This may allow the control over the molecular structure design and surface selectivity. The incorporation of hydrophilic domains may increase the BCPs selectivity, enhance the phase segregation and facilitate transport properties. The addition of hydrophobic domains may improve the mechanical properties of the polymers. The BCPs may then be placed on electrodes. The proposed polymers are conductive polymers with no interference to the Raman signal of the biomarker, providing various detection alternatives to the biomarkers.

[0034] Accordingly, the biosensor 100 may provide a low-cost, portable, and fast diagnostic test for heart attacks. Compared to existing tests, the biosensor 100 may be noninvasive, for example sensing cTnl in sweat or saliva. Further, the biosensor 100 may be more accurate than current tests for myocardial infarction. For example, certain existing tests detect cardiac troponin T, which is expressed in small amounts in skeletal muscle and thus raises risk of incorrect diagnosis. In contrast, the biosensor detects cTnl, which is unique to the myocardium. Additionally, the biosensor 100 may be manufactured from inexpensive materials or otherwise have reduced costs compared to existing tests. For example, unlike certain existing tests, the disclosed biosensor 100 does not require refrigeration to prolong test viability. Further,Docket No. 23-002-UPR / 71900-431712 as described below, the biosensor 100 may be integrated into a wearable device, which may allow for continual monitoring for myocardial infarction. Continual monitoring may provide for earlier detection. Improving detection speed may improve patient outcomes by allowing for faster treatment.

[0035] As best shown in FIG. 2, the illustrative sensor 100 includes a base 1 12, which may be formed from poly(methyl methacrylate) (PMMA) or another appropriate structural material. The base 112 is covered with a non-conductive polymer film substrate 114, which is illustratively formed from polyimide (PI). Additionally or alternatively, in some embodiments the substrate 114 may be formed from another material, such as poly(styrene-isobutylene- styrene), poly(styrene-ethylene-butylene-styrene), poly((styrene-ethylene-styrene), and / or graphene / graphene oxide. The electrodes 102 are laser-scribed electrodes 116 formed on the surface of the substrate 114. The laser-scribed electrodes 116 are formed from a conductive carbon-based material which may include or be similar to graphite, graphene, or another carbonbased material.

[0036] The film substrate 114 and electrodes 116 are covered by an electrode cover 118. As shown, the cover 118 includes one or more openings 120 that, when the cover 118 is positioned on the substrate 114, expose part or all of the electrodes 116. Similarly, the cover 118 may be covered by a cap 122 that includes an opening 124 that aligns with the openings 120. Each of the cover 118 and the cap 122 may be formed from a semi-rigid polymeric material such as vinyl. Additionally or alternatively, in some embodiments the cover 118 and / or the cap 122 may be formed from any thin polymer that may be cut, such as acrylic, polystyrene, polycarbonate, polypropylene films or thin sheets. The cover 118 and the cap 122 may encapsulate and protect the components of the sensor 100 while allowing part or all of the electrodes 116 to be exposed to the environment for sensing. Both the cover 118 and the cap 122 help to control the electrolyte (e.g., ferricyanide) volume deposited on the electrodes 116, which may be used to sense and measure Electrochemical Active Surface Area (EASA) and Heterogeneous Electron Transfer (HET) in different manufacturing conditions.

[0037] Referring now to FIG. 3, an illustrative embodiment of a wearable device 300 of the present disclosure is depicted. As shown, the illustrative wearable device 300 is a smart watch. The illustrative smart watch 300 includes a body case 302 and two wrist straps 304, 306. FIG. 3 illustrates a bottom surface 308 of the case 302. A biosensor 100 including exposed electrodes 102 as shown in FIGS. 1-2 is positioned on the bottom surface 308 of the case 302. Accordingly, when the watch 300 is attached to a user’s wrist, the electrodes 102 of the biosensor 100 are placed in contact with the user's skin. Accordingly, the sensor 100 may detect biomarkersDocket No. 23-002-UPR / 71900-431712 present in the user’s sweat or other biomarkers present on the user’s skin. The device 300 includes a controller 310 which may be coupled to the biosensor 100. The controller 310 may execute one or more control routines or other algorithms for detecting biomarkers using the biosensor 100. One potential embodiment of a method for detecting the biomarker is described further below in connection with FIG. 4.

[0038] The controller 310 may be embodied as any type of microcontroller, microprocessor, or compute engine capable of performing the functions described herein. For example, the controller may be embodied as a single or multi-core processor(s), digital signal processor, microcontroller, or other processor or processing / controlling circuit. The wearable device 300 may further include other or additional components, such as those commonly found in a smartwatch, smartphone, or other smart device, (e.g., a memory, data storage, various input / output devices, and other components).

[0039] Although illustrated as a smartwatch 300, it should be understood that in other embodiments the sensor 100 may be included in other types of wearable devices, wearable biosensors, and / or other devices include wearable features, such as a wristband, a watch, a bracelet, gloves, clothing, an adhesive patch, or other devices. Additionally or alternatively, in some embodiments the sensor 100 may be included in an implant or other implantable device.

[0040] Referring now to FIG. 4, in some embodiments the biosensor 100 and / or the wearable device 300 may be used to perform a method 400 for detection of a health-related biomarker. The method 400 begins in block 402, in which the biosensor 100 is placed in contact with a specimen. The specimen may be embodied as a material that may include the health- related biomarker, such as sweat or saliva of a user. In some embodiments, the biosensor 100 may be included in or otherwise coupled to a wearable device 300 that is attached to the user. Thus, the biosensor 100 may be in contact with a specimen on the user’s skin. In some embodiments, the specimen may be a blood sample from the user.

[0041] In block 404, an electrochemical response of the biosensor 100 is detected. For example, a controller 310 may control the biosensor 100 and detect the electrochemical response of the biosensor 100. Additionally or alternatively, in some embodiments, the biosensor 100 may be coupled to a potentiostat or other instrument capable of detecting the electrochemical response of the biosensor 100. In some embodiments, in block 406 cyclic voltammetry may be applied to the biosensor 100. In some embodiments, in block 408 linear sweep voltammetry may be applied to the biosensor 100. In some embodiments, in block 410 differential pulse voltammetry may be applied to the biosensor 100.Docket No. 23-002-UPR / 71900-431712

[0042] In block 412 it is determined whether a peak indicative of a health-related biomarker was detected in the electrochemical response of the biosensor 100. For example, the controller 310 or other device may determine whether the voltammetry data generated as described above in connection with block 404 includes a current peak at a particular predetermined voltage or range of voltages. If no peak is detected, the method 100 loops back to block 402 to continue monitoring for the biomarker. If a peak was detected, the method 400 advances to block 414.

[0043] In block 414, detection of the health-related biomarker is indicated. For example, a controller 310 of a wearable device 300 may display an indication that the health-related biomarker is detected, sound an alarm, generate haptic feedback, or otherwise indicate detection of the health-related biomarker. After indicating detection of the biomarker, the method 400 loops back to block 402 to continue monitoring for the biomarker.

[0044] Referring now to FIG. 5, chart 500 illustrates experimental results that may be achieved with a biosensor 100 according to FIGS. 1 -4. In an experiment, a biosensor 100 including screen-printed electrodes 102 with SIBS block copolymer was placed into contact with a solution of 10 pL of cTnl diluted in 30 pL of commercial PBSxl. Cyclic voltammetry data for the biosensor 100 was recorded for multiple scan rates. Cyclic Voltammetry measurements were taken at potential sweeps from -500 to 500 mV at different scan rates (illustratively, 10, 20, 50, 75, 100, 150, 200, 500 mV / s) to calculate the Electrochemical Active Surface Area (EASA) and Heterogeneous Electron Transfer (HET) rate using 50 pl of potassium ferricyanide 3.3 mM in 0.1 M KC1. In the illustrative experiment, electrode outputs were measured using Cyclic Voltammetry with a Gamry Series G300 potentiostat (Gamry Instruments, Warminster, PA, USA).

[0045] Accordingly, each curve shown in the chart 500 represents voltammetry data recorded at a particular scan rate. As shown, a peak 502 is present in the voltammogram for each scan rate. This peak 502, which illustrative occurs at 0.05 ± 0.005 V in all scan rates, represents the presence of cTnl. Accordingly, when cTnl is not present, the peak 502 is not detected in voltammetry data.

[0046] Referring now to FIG. 6, in some embodiments a method 600 for manufacturing a biosensor 100 may be performed. The method 600 begins in block 602, in which one or more electrodes 102 are provided. In some embodiments, in block 604 a screen-printed electrode (SPE) may be provided. In some embodiments, in block 606 a laser-scribed electrode (LSE) may be provided. The SPE may be a commercially available SPE, such as an SPE available from BVTDocket No. 23-002-UPR / 71900-431712Technologies, Strazek, Czech Republic. Potential embodiments of methods for manufacturing a LSE are described further below in connection with FIGS. 8 and 9.

[0047] In block 608, a sulfonated block copolymer is created. As discussed above, block copolymers (BCPs) are a type of polymer with potential adsorption sites for biomarkers. The proposed polymers are conductive polymers with no interference to the Raman signal of the biomarker, providing various detection alternatives to the biomarkers. Some advantages of BCPs include the possible functionalization of specific functional groups, that may improve the physical and chemical properties of the polymer. This may allow the control over the molecular structure design and surface selectivity. The incorporation of hydrophilic domains may increase the BCPs selectivity, enhance the phase segregation and facilitate transport properties. The addition of hydrophobic domains may improve the mechanical properties of the polymers.

[0048] In some embodiments, in block 610 the sulfonated block copolymer is sulfonated styrene-isobutylene-styrene (SIBS) block copolymer. In an embodiment, a sulfonating agent was prepared. The sulfonating agent with a molar ratio of 3:1 of acetyl sulfate to polystyrene was prepared by the addition of 60.00 mL of 1,2-dichloromethane (DCM) in an Erlenmeyer flask. The solution was cooled below 5°C. 6.87 mL of acetic anhydride was added in a round flask with DCM and 3.86 mL of 98% sulfuric acid was added slowly keeping temperature below 5°C. A clear solution resulted. The acetyl sulphate reaction mechanism is described below.

[0049] In a three neck round flask, 10 g of pre-dried (i.e., 60 °C for 24 h) SIBS were added and dissolved in methylene chloride to obtain a 10% (wt. / v) solution. Acetyl sulfate (sulfonating agent) was produced from the reaction of sulfuric acid and acetic anhydride in methylene chloride at 0 °C. The sulfonating agent was slowly added to the polymer solution to begin the sulfonation and stopped after 24 h with methanol. The solvents were allowed to completely evaporate at room temperature, 2-3 days. The functionalized polymer was washed with (DI) water until the solution reached a pH close to 7. The mole ratio of acetyl sulfate to polystyrene was 3:1 for this study, respectively. Referring quickly to FIG. 7, diagram 700 presents the sulfonation synthesis mechanism of SIBS block copolymer. The sulfonation mole percent of the block copolymers was determined from Elemental Analysis (EA). The sulfonation levels obtained were 85 mol% for a mole ratio of 3: 1 (acetyl sulfate: polystyrene).

[0050] In block 612 of FIG. 6, the sulfonated block copolymer 110 is deposited on one or more of the electrodes 102. For example, the sulfonated block copolymer 110 may be deposited on the working (W) electrode 106 of a three-electrode sensor as illustrated in FIGS. 1Docket No. 23-002-UPR / 71900-431712 and 2. In an embodiment, after the sulfonating block copolymer characterization, a STBS solution was prepared with a concentration of 20 mg / mL dissolved in chloroform (98%). Then, 1 pL of the polymer solution was added to the SPE carbon surface (center) and dried at 60 °C for 3 hours. After depositing the sulfonated block copolymer 110, the biosensor 100 is completed. The biosensor 100 may be used as described above in connection with the method 400 shown in FIG. 4 to detect a health-related biomarker such as cTnl. Additionally or alternatively, in some embodiments, the biosensor 100 may be incorporated into a wearable device 300 or other device to perfonu detection of the health-related biomarker. The electrode may be characterized

[0051] Referring now to FIG. 8, in some embodiments a method 800 for manufacturing a laser-scribed electrode (LSE) may be performed. The method 800 begins in block 802, in which a nonconductive polymer film substrate 114 is provided. In some embodiments, in block 804 the film substrate 114 may be formed from polyimide. As described above, in other embodiments the substrate 114 may be formed from other materials, such as such as poly(styrene-isobutylene- styrene), poly(styrene-ethylene-butylene-styrene), poly((styrene-ethylene-styrene), and / or graphene / graphene oxide.

[0052] In block 806, one or more electrodes 116 are scribed on the surface of the film substrate 114 with a laser. In an embodiment, laser-scribed carbon-based electrodes (LSEs) were manufactured using an Epilog Mini 12x24 60- Watt CO2 laser cutter (Epilog Corporation, CO, USA) inscribed on Polyimide film (Polyimide Kapton Film, 5 mil thickness). The illustrative LSEs had dimensions of 2 cm in length, and 1.4 mm (4 pt.) or 0.7 mm (2 pt.) width. After applying the laser to the surface of the film 114, a carbon-based electrode layer 116 is formed on the surface of the film 1 14. In an experiment, Raman spectrometry was used to investigate the material structure of the laser scribed carbon-based electrodes 116. In the illustrative embodiment, the carbon-based electrodes 116 had a structure more similar to graphite than to graphene. In block 808, one or more parameters of the laser may be configured prior to scribing the electrodes. The laser uses different manufacturing parameters; illustratively, the manufacturing parameters may include Speed, Power, Focus, and Dots per Inch (DPT). In an embodiment, the LSE was manufactured using the best identified settings: speed 25%, power 12% and width 4 pt. Changing the laser parameters may change the structure of the carbon-based electrodes 116. For example, higher speed and lower power for the laser may result in less thermal ablation, which may be used to create carbon structures with fewer defects or disorders in order to yield a material closer to graphene. One potential embodiment of a method for optimizing the parameters of the laser is described below in connection with FIG. 9.Docket No. 23-002-UPR / 71900-431712

[0053] In block 810, the film substrate 1 14 is mounted to a base 1 12. As described above, the base 112 may be formed from a structural material such as PMMA. In block 812, the film substrate 114 is covered with a cover 118 and / or a cap 122. Each of the cover 118 and the cap 122 includes one or more respective openings 120, 124. When positioned on the film substrate 1 14, those openings 120, 124 are aligned with part or all of the laser-scribed electrodes 116. Accordingly, the laser-scribed electrodes 116 may contact a specimen through those openings 120, 124. After covering the film substrate 114, manufacturing of the electrodes 102 may be completed. As described above in connection with FIG. 6, after the laser-scribed electrode 102 is manufactured, a sulfonated block copolymer 110 may be deposited on one or more of the electrodes 102. Although illustrated separately, in some embodiments the operations of the methods 600, 800 may be performed together and / or in a different sequence. For example, in some embodiments, the sulfonated block copolymer 110 may be deposited onto the laser-scribed electrodes 116 prior to the cover 118 and / or the cap 122 being attached over the film substrate 1 14. Therefore, it may be important to identify the capability of stable equilibrium potential in the reference electrode in both since a drifting potential can have many undesired effects.

[0054] Referring now to FIG. 9, in some embodiments a computing device or other computing system may perform a method 900 for optimizing laser manufacturing parameters for a laser-scribed electrode (LSE). The method 900 begins in block 902, in which the computing device trains a generator model and a discriminator model with electrical parameters for laser- scribed electrodes that were produced with various manufacturing parameters. Generative Adversarial Networks (GAN) is a deep learning method used to automatically identify and learn the patterns in the input data so that the model can produce new examples that could have been reasonably derived from the original dataset. GANs comprise two sub-models: the generator model, which is trained to create new instances, and the discriminator model, which tries to categorize examples as either real or fake. This methodology is being used successfully in scenarios that suffer from data scarcity. In this disclosure, GAN is used to generate new manufacturing settings that can be investigated further experimentally. This approach may reduce the cost and time required to explore the whole domain of potential manufacturing settings. For graphene-based electrode manufacturing (e.g., laser-scribed electrodes (LSEs)), the GAN generator produces experimental manufacturing settings where the laser cutter power and speed parameters are variables and seeks to maximize the Electrochemical Active Surface Area (EASA) and the Heterogeneous Electron Transfer (HET) as a function of the cyclic voltammetry oxidation. This conceptualization is used to design scores for the potential experimental manufacturing setting to achieve the best electrical recordings while achieving as close to singleDocket No. 23-002-UPR / 71900-431712 layer graphene material as possible. Tn an embodiment, the data may be represented as RGB images containing EASA, HET, and speed parameters.

[0055] Training data may be based on measurements of manufactured laser-scribed electrodes. For example, electrochemical active surface area (EASA) and heterogenous electron transfer (HET) may be determined based on voltammetry measurements of manufactured laser- scribed electrodes. Continuing that example, in an experiment, cyclic voltammetry measurements were taken at potential sweeps from -500 to 500 mV at different scan rates (10, 20, 50, 75, 100, 150, 200, 500 mV / s) to calculate the Electrochemical Active Surface Area (EASA) and Heterogeneous Electron Transfer (HET) rate using 50pl of potassium ferricyanide 3.3 mM in 0.1 M KC1. Approximations of the EASA were obtained from the Randles-Sevcik equation:Ip= 26.86xl04n3l2Dll2v CA (1) where Ip (Amperes) is the reduction current, n is the number of electrons contributing to the redox reaction (n=l), D (cm2 / s) is the diffusion coefficient (6.7xl0-6cm2 / s for K3[Fe(CN)6]4-), v (V / s) is the scan rate, C (mol / cm3) is the concentration of the probe molecule and A is the estimated EASA. The HET rate (kO) was estimated via Nicholson’s treatment: ip = k° [— (2) where R is the universal gas constant (8.314 J / mol*K), F is the Faraday constant (96485 C / mol), T is the absolute temperature and p is the dimensionless kinetic parameter. Since this experiment entails a one electron process (n=l), ip depends on the peak-to-peak separation (AEp) and can be obtained from the following equation:where X represents AEp x n. Through the integration of Equations 2 and 3, can be determined from the slop ' e of ip versus

[0056] In block 904, the computing device uses the generator model to generate candidate manufacturing parameters for laser-scribed electrodes. In some embodiments, in block 906 the manufacturing parameters may include laser cutter power, speed, focus, dot pitch, or other control parameters.

[0057] In block 908, the computing device uses the discriminator model to select manufacturing parameters from the candidate parameters to achieve maximum electrical properties. In some embodiments, in block 910 the discriminator model may be used to maximize electrochemical active surface area (EASA) and heterogenous electron transfer (HET).Docket No. 23-002-UPR / 71900-431712

[0058] In block 912, the selected manufacturing parameters may be tested, and training data for the generator model and the discriminator model may be updated. For example, in an embodiment, one or more LSEs may be manufactured using the selected manufacturing parameters, and the electrochemical response of the manufactured LSEs may be measured. The training data may be updated with the measured data for the manufactured LSEs, given the selected manufacturing parameters. After updating training data, the method 900 loops back to block 902 to continue optimizing the manufacturing parameters using generative adversarial networks as described above.

[0059] All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While this invention may be embodied in many different forms, there are described in detail herein specific preferred embodiments of the invention. The present disclosure is an exemplification of the principles of the invention and is not intended to limit the invention to the particular embodiments illustrated. In addition, unless expressly stated to the contrary, use of the term “a” is intended to include “at least one” or “one or more.” For example, “a sulfonated block copolymer” is intended to include “at least one sulfonated block copolymer” or “one or more sulfonated block copolymers.”

[0060] Any ranges given either in absolute terms or in approximate terms are intended to encompass both, and any definitions used herein are intended to be clarifying and not limiting. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Moreover, all ranges disclosed herein are to be understood to encompass any and all subranges (including all fractional and whole values) subsumed therein.

[0061] Any composition disclosed herein may comprise, consist of, or consist essentially of any element, component and / or ingredient disclosed herein or any combination of two or more of the elements, components or ingredients disclosed herein.

[0062] Any method disclosed herein may comprise, consist of, or consist essentially of any method step disclosed herein or any combination of two or more of the method steps disclosed herein.

[0063] The transitional phrase “comprising,” which is synonymous with “including,” “containing,” or “characterized by,” is inclusive or open-ended and does not exclude additional, un-recited elements, components, ingredients and / or method steps.Docket No. 23-002-UPR / 71900-431712

[0064] The transitional phrase “consisting of’ excludes any element, component, ingredient, and / or method step not specified in the claim.

[0065] The transitional phrase “consisting essentially of’ limits the scope of a claim to the specified elements, components, ingredients and / or steps, as well as those that do not materially affect the basic and novel characteristic(s) of the claimed invention.

[0066] As used herein, the term “about” refers to the cited value being within the errors arising from the standard deviation found in their respective testing measurements, and if those errors cannot be determined, then “about” may refer to, for example, within 5% of the cited value.

[0067] Furthermore, the invention encompasses any and all possible combinations of some or all of the various embodiments described herein. It should also be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the invention and without diminishing its intended advantages. It is therefore intended that such changes and modifications be covered by the appended claims.

Claims

Docket No. 23-002-UPR / 71900-431712WHAT IS CLAIMED IS:

1. A wearable device comprising: a biosensor comprising a plurality of electrodes; a functionalized coating coupled to a first electrode of the plurality of electrodes, wherein the functionalized coating comprises a sulfonated block copolymer; a controller coupled to the biosensor; and a wearable feature coupled to the biosensor; wherein the controller is configured to (i) sense an electrochemical response of the biosensor and (ii) determine whether a health-related biomarker is present based on the electrochemical response.

2. The device of claim 1, wherein the sulfonated block copolymer comprises a sulfonated styrene-isobutylene-styrene (SIBS) block copolymer.

3. The device of claim 1 or claim 2, wherein the health-related biomarker comprises cardiac troponin I (cTnl).

4. The device of any one of the preceding claims, wherein to sense the electrochemical response of the biosensor comprises to apply electrochemical impedance spectroscopy, cyclic voltammetry, linear sweep voltammetry, or differential pulse voltammetry to the biosensor.

5. The device of any one of the preceding claims, wherein the plurality of electrodes comprises a screen printed electrode.

6. The device of any one of the preceding claims, wherein the plurality of electrodes comprises gold or silver.Docket No. 23-002-UPR / 71900-4317127. The device of any one of the preceding claims, wherein the plurality of electrodes comprises a conductive carbon-based polymer.

8. The device of any one of the preceding claims, wherein the plurality of electrodes comprises laser-scribed electrodes.

9. The device of any one of the preceding claims, wherein the wearable feature comprises a strap, an adhesive patch, a glove, an article of clothing, or an implant.

10. A method for manufacturing a biosensor, the method comprising: providing a film substrate: scribing a plurality of electrodes on a surface of the film substrate with a laser, wherein the laser is configured with first manufacturing parameters: mounting the film substrate to a base; and positioning a cover on the film substrate after scribing the plurality of electrodes, wherein the cover comprises an aperture that when positioned on the film substrate exposes at least part of each of the plurality of electrodes.

11. The method of claim 10, wherein the first manufacturing parameters comprise laser power, speed, laser focus, and / or laser dot pitch.

12. The method of claim 10 or claim 11, wherein the film substrate comprises a non- conductive polymer film substrate.

13. The method of any one of the preceding claims, wherein the film substrate comprises a polyimide (PI) film substrate.

14. The method of any one of the preceding claims, wherein the base comprises a poly(methyl methacrylate) (PMMA) base.Docket No. 23-002-UPR / 71900-43171215. The method of any one of the preceding claims, wherein the cover comprises a semi-rigid polymeric material.

16. The method of any one of the preceding claims, wherein the cover comprises a vinyl cover.

17. The method of any one of the preceding claims, further comprising: depositing a SIBS block copolymer on a first electrode of the plurality of electrodes.

18. A method for manufacturing a biosensor, the method comprising: training, by a computing device, a generator model and a discriminator model with training data, the training data comprising electrical parameters for laser-scribed electrodes associated with manufacturing parameters; generating, by the computing device, a plurality of candidate manufacturing parameters with the generator model in response to training the generator model: selecting, by the computing device, a first candidate manufacturing parameter of the candidate manufacturing parameters with the discriminator model in response to training the discriminator model; and manufacturing a test biosensor with the first candidate manufacturing parameter.

19. The method of claim 18, further comprising: evaluating one or more electrical parameters of the test biosensor; and updating, by the computing device, the generator model and the discriminator model with the first candidate manufacturing parameter and the electrical parameters of the test biosensor.

20. The method of claim 18 or claim 19, wherein the manufacturing parameters comprise laser power, speed, laser focus, and laser dot pitch.

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