Electrochemical sensor for non-invasive detection

Abstract

A device for sensing is provided. A sensing layer having a three dimensional dispersion of aminated graphene in a conductive polymer matrix is on a support layer. The matrix has cavities on the surface formed by self-assembled chemistry and which are tailored binding sites to a template molecule, and the cavities are surrounded by dopamine and aminated graphene.

Description

[0001] ELECTROCHEMICAL SENSOR FOR NON-INVASIVE DETECTION

[0002] CROSS-REFERENCE TO RELATED APPLICATION

[0003] (001) This application claims the benefit of U.S. Provisional Patent Application Serial No. 63 / 534,730 filed August 25, 2023, which is hereby incorporated by reference in its entirety.

[0004] GOVERNMENT SUPPORT.

[0005] (002) This invention was made with U.S. government support under 75A50121C00075 awarded by Biomedical Advanced Research and Development Authority (ASPR / BARDA). The government has certain rights in this invention.

[0006] FIELD

[0007] (003) This disclosure relates generally to molecular sensors. More specifically, this disclosure relates electrochemical sensors for sensing template molecules.

[0008] BACKGROUND

[0009] (004) Early and timely detection of such as disease biomarkers and other template molecules may prevent the spread of the diseases and drastically decrease death. Because conventional detection and diagnostic methods have limited application in low-resource settings due to bulky and expensive instrumentation, low-cost and simple point-of-care detection and diagnostic devices for timely and early biomarker diagnosis are the need of the hour. Most existing technologies are limited because of a lack of robustness, multiple assay steps and scalability.

[0010] SUMMARY

[0011] (005) According to one aspect of the disclosed subject matter, a device for sensing is provided. A sensing layer having a three dimensional dispersion of aminated graphene in a conductive polymer matrix is on a support layer. The matrix has cavities on the surface formed by self-assembled chemistry and which are tailored binding sites to a template molecule, and the cavities are surrounded by dopamine and aminated graphene. (006) Tn another embodiment, a device for sensing is provided. A sensing layer is comprised of molecularly imprinted polymer (MIP) on a supporting layer. The molecularly imprinted polymer (MIP) having a 3D matrix of graphene assembled from one or more layers of nanotextured reduced graphene with NH2, NH3, OH, O, and COOH, functionally forming a 3D matrix of graphene dispersed within the MIP. The one or more layers of reduced nanotextured graphene form a p-p isotype with the molecularly imprinted polymer (MIP).

[0012] (007) These and other aspects of the disclosed subject matter, as well as additional novel features, will be apparent from the description provided herein. The intent of this summary is not to be a comprehensive description of the claimed subject matter, but rather to provide a short overview of some of the subject matter's functionality. Other systems, methods, features and advantages here provided will become apparent to one with skill in the art upon examination of the following FIGUREs and detailed description. It is intended that all such additional systems, methods, features and advantages that are included within this description, be within the scope of any claims.

[0013] BRIEF DESCRIPTION OF THE FIGURES

[0014] (008) The features, natures, and advantages of the disclosed subject matter may become more apparent from the detailed description set forth below when taken in conjunction with the drawings in which like reference numerals indicate like features.

[0015] (009) Figure 1 is an expanded portion diagram of the sensor of Figure IB in accordance with the disclosed subject matter.

[0016] (0010) Figure IB is a diagram of a sensor in accordance with the disclosed subject matter.

[0017] (0011) Figure 1C is a diagram showing positive charge (hole) movement from graphene to polypyrrole (PPy) in accordance with the disclosed subject matter.

[0018] (0012) Figures 2A, 2B, and 2C are graphs showing the sensitivity testing results of the template molecule (CXCL-13) sensor in accordance with the disclosed subject matter.

[0019] (0013) Figure 3A is a diagram showing an enzyme-linked immunosorbent assay (ELISA). (0014) Figure 3B is a graph showing a comparison of the template molecule (CXCL-13) sensor in accordance with the disclosed subject matter and an enzyme-linked immunosorbent assay (ELISA).

[0020] (0015) Figure 4 is a graph showing the testing results of a template molecule (IL-6 cytokine) sensor in accordance with the disclosed subject matter.

[0021] DESCRIPTION

[0022] (0016) The following description is not to be taken in a limiting sense, but is made for the purpose of describing the general principles of the present disclosure. The scope of the present disclosure should be determined with reference to the claims. The features and advantages of the embodiments will be more readily appreciated when considered in conjunction with the accompanying figures and drawings. Exemplary embodiments of the present disclosure may be illustrated in the drawings, like elements being used to refer to like and corresponding parts of the various drawings. The dimensions of drawings provided are not shown to scale.

[0023] (0017) Molecular imprinting is a technique in which a thin polymer layer is electrochemically formed around the template molecules of interest (i.e., the template molecule is the molecule to be detected or the molecule of interest). The template molecules are then removed after polymerization and it leaves an imprint of the exact size and shape of the template molecule in the polymer matrix. This results in a high specificity lock (sensor cavity) and key (template molecule) relationship. In addition, the strong chemical bonding between the template molecules and the electrochemically plated polymer layer leads to further enhanced selectivity.

[0024] (0018) Three critical elements in molecular imprinting technique include: the target molecule (or template) which is what is being sensed; the functional monomer, which is a compound having chemical and shape complementarity to the template that will help the polymerization to form the polymer matrix; and the cross-linking agent (or cross-linker) that is a multifunctional molecule containing two or more reactive split ends able to interact via chemical bound.

[0025] Graphene is a 2D material known for its unique electrochemical and structural properties. One atom thick graphene has high thermal and electrical conductivity, chemical stability, high specific surface area, and excellent room temperature carrier mobility. Prussian blue (PB, ferric ferrocyanide), formula Fe4 [Fe(CN)6]3, is a microcrystalline blue powder. It is an inorganic conductive material that exhibits high electrocatalytic activity and improves electrochemical sensitivity. It has been extensively studied for its attractive electrochemical properties and wide applications in the field of sensors. In many cases, PB has been used as a mediator with graphene. The combination of graphene and PB leads to a synergistic effect that can help the electron transfer. MIPs are widely used in various fields such as purification, separation, and catalysis, and degradation processes but also they have become attractive in drug delivery, artificial antibodies, or biosensing. Past MIP-based sensors were created on rigid substrates. However, if a highly sensitive and specific sensor could be integrated into flexible materials, it could have great value for medical, industrial, and environmental sensing applications (e.g., artificial blood vessels, gas line valves, and wearable clothing with integrated chemical detection functionalities).

[0026] (0019) The innovations described provide an innovative solution for an advantageously robust sensor that is entirely non-invasive, super sensitive, and highly selective. The sensor has an immediate response (less than 10 seconds) and fast recovery as well as being reproducible, cost-effective, accurate, and reliable. As this sensor system enables rapid diagnosis with only minimal user skills, it may be useful for diagnosis at home and in pharmacies, ambulances, doctor offices, and hospitals. Compared to other available products and technologies, the sensor and sensing method disclosed may be fabricated on a large scale (up to at least 10,000 sensors) in one day.

[0027] (0020) Described herein is a new type of molecularly imprinted polymer (MlP)-based electrochemical sensor that utilizes a 3D nanocomposite with imprinted binding sites within and throughout its depth, notably in contrast to the layered method used in the past. In addition, as described herein, the application of this electrochemical sensor creates an elastomeric MIP-based electrochemical sensor that may be used in highly sensitive chemical detectors in wearable materials or in flexible materials for industrial or medical applications.

[0028] (0021) Structure Detail. Exemplary structural details are provided below, as described with reference to CXCL-13 as the template molecule. (0022) An electrochemical gas sensor for CXCL-13, a cytokine biomarker of inflammation, has been developed and tested. The electrochemical gas sensor is composed of a graphene / PB nanocomposite containing molecularly imprinted polymeric (polypyrrole) nanofibers immobilized on a silicon substrate that is used to detect template molecules such as biomarker molecules with high sensitivity and specificity based on their ability to bind into the imprinted molecular pocket. MIPS, often referred to as synthetic antibodies, offer an alternative approach to biomolecular and viral recognition.

[0029] (0023) The diagrams of Figures 1A, IB and 1C show the MIP cavities surrounded by dopamine molecules which are electron donors. Figure 1A is an expanded portion diagram of the sensor of Figure IB. Figure 1C is a diagram showing positive charge (hole) movement from graphene to polypyrrole (PPy). Template molecule CXCL13 is positively charged and once the template molecule CXCL13 is captured (i.e., trapped) in the cavity, it can bind with dopamine (hydrogen bonding) that can decrease the charge in polypyrrole (PPy). In operation, the graphene (Aminated Graphene in Figure 1 A) is mainly acting as a transducer while the main interaction is primarily located at the PPy. Indeed, the potential applied to the transducer element (graphene) is increasing the charge flow and in consequence favoring the hole movement from graphene to PPy over the barrier. Binding on the surface causes graphene to flow majority charge carriers (holes) toward PPy. This will decrease the conductivity of graphene leading to increase in resistance of the sensor. The p-p (positive-positive) isotype junction, the junction between graphene and polypyrrole (PPy) that are formed around each cavity, is at equilibrium under ideal conditions, and it shows greater discontinuity in the conduction bands (AEC) relative to the valence bands (AEV). Therefore, excitations of the sensitive layer translate into positive charges (holes) being emitted from the graphene over the barrier into the PPy. In consequence, near the boundary of the interface a depletion region can be formed, specifically located on the graphene surrounding PPy. Conversely, PPy likely increases carrier concentration, thus being considered accumulation regions. As a result, once the cytokine donates electrons to the PPy@Graphene layer, the Fermi level shifts upward. This shift in the Fermi level for graphene and PPy is associated with a band-bending readjustment at the time of exposure.

[0030] (0024) On the other hand, molecules other than CXCL13 (e.g., cytokines: CXCL10, MIP-1B,

[0031] IL-4) that are positively charged can get close to the sensing layer but they do not get captured (i.e. , trapped) by the cavity. In such cases, the molecule's positive charges would repel the excess positive charge of PPy toward graphene that would lead to lowering the graphene resistance. This helps explain the decrease in the resistance while exposed to other cytokines.

[0032] (0025) The diagrams of Figures 1A, IB and 1C show the mechanism of sensing. Notably, the technology works based on self-assembled chemistry. Once the template molecule such as biomarker, CXCL13 in this case, is trapped (i.e., captured) inside the cavity which is formed by polypyrrole and encapsulated by polydopamine and graphene, the Fermi level of graphene changes and the conductivity of the sensor decreases. In other words, as described with reference to the diagrams of Figures 1A, IB and 1C and the fabrication detail herein, the cavities are formed by self-assembled chemistry of the template molecule and dopamine on the three dimensional (3D) matrix of graphene and polypyrrole. By later removing the template molecule, these cavities will be the tailored binding sites for the template molecules. Thus, the cavities are formed by self-assembled chemistry. And a Sensing Layer which has 3D dispersion of aminated graphene (Aminated Graphene in Figure 1 A), in a polypyrrole matrix with cavities on the surface which are tailored binding sites to the template molecule — the molecule to be detected — is formed. Aminated graphene refers to amine functionalized graphene having some amine group(s) attached to the graphene, for example pentaminated graphene having five amine groups attached to the graphene. These Sensing Layer cavities are surrounded by dopamine (Dopamine as electron donor in Figure 1A) and aminated graphene. Thus, the molecular imprinted surfaces provide geometric similarity (i.e., the surfaces are tailored) to the template molecule (e.g., virus, etc.) while the self-assembled chemistry provides chemical attraction to the template molecule, thus providing the best of both worlds to attract and capture a template molecule (e.g., virus, etc.).

[0033] (0026) The conductivity of amine group attached to graphene varies widely based on type and number of amine group attached to graphene. However, aminated graphene, for example pentaminated graphene, has a lower electrical conductivity than pure graphene. And as used herein, the unexpectedly advantageous aminated graphene, pentaminated graphene, may provide improved chemical binding with the molecules of interest.

[0034] (0027) Fabrication Detail. Exemplary fabrication details are provided below. (0028) Fabrication steps are as follows: a. A highly doped double side polished (DSP) premium silicon wafer was used. b. The wafer was then etched with hydrofluoric acid (HF) to remove the SiO2 layer altogether. c. A thick layer of photoresist was deposited on the surface using a spinner to prevent the formation of SiO2 on the surface. d. The wafer was then diced into 7X7 mm2 (0.27x0.27 inch2). Each diced piece is one sample. e. A sample was washed with acetone followed by isopropyl alcohol and water to remove the photoresist completely. f. Preparation of graphene / PB: 6 mg of pentaminated reduced graphene oxide (rGO) was added to 2 mL of deionized water and ultra-sonicated to obtain a homogenous black solution. The solution was then added into 5 mL of an aqueous solution containing 0.006 g FeC13.6H2O, 0.008 g K3Fe(CN)6, and 0.037 g KC1 by stirring at room temperature for 12 hours. The product was then centrifuged (2000 rpm for 20 min), washed with the deionized water several times, and dried overnight under a vacuum at 40°C. As a result, powder of rrGO / PB is obtained. g. 100 ml of phosphate buffer solution was poured into a beaker, and the pH was adjusted to 6.8 using pure sulfuric acid. h. 693 ul of pyrrole (monomer) was mixed with 150 ul of rGO / PB mixture and ultra-sonicated for 30 minutes. i. 18 ul of already purified CXCL-13 (template molecule) was mixed with 10 mg of dopamine hydrochloride in 1 ml of phosphate buffer solution and placed on the rotation table for 1 hour. Dopamine in a slightly basic solution (pH above 7) forms a thin layer of polydopamine around any object inside the solution. Dopamine was used as a functional monomer and the purpose was to prepolymerized before the polymerization step. j . The solutions prepared in steps 7 to 9 were mixed and stirred for 1 minute (electrolyte solution). k. The working electrode (silicon sample), the counter electrode (Platinum mesh), and the reference electrode (Ag / Agcl) were placed in the electrolyte solution and connected to a potentiostat. l. Sensor preparation: cyclic voltammetry in the range of -0.5 to +0.5 V, speed: 50 mV / s were performed for 20 cycles to form a thin layer of polypyrrole with template molecule to be formed on the surface of the sample (molecular imprinting polymer). m. The sensor was then washed with 70% ethanol to remove the template molecule and stored overnight.

[0035] (0029) Results. Testing results of the sensor fabricated above are provided below.

[0036] (0030) The CXCL-13 gas sensor was tested against both its template molecule (CXCL-13) and other similar cytokines as shown in the graphs of Figures 2A, 2B, and 2C. Figures 2A, 2B, and 2C are graphs showing the sensitivity testing results of the template molecule (CXCL-13) sensor. The testing procedures is briefly described below: a. In an Eppendorf tube, the cytokine of interest was diluted in Roswell Park Memorial Institute (RPMI) culture medium. b. The sensor was tested for 30 seconds, 10 seconds of stabilizing, 10 seconds of exposure, and another 10 seconds of post-exposure stabilizing. c. The Eppendorf tube was opened and placed just under the sensor surface without touching the sensor during the exposure. d. The sensor's resistance was monitored with a digital multimeter during the test that was connected to metal contacts on the sensing layer. This may also be accomplished by a four point probe method such that the probe tip is metallized for electrical contact.

[0037] (0031) Sensitivity: The sensor was tested with 5 samples with CXCL-13 concentration between 1 to 20 pg / ml. The sensor showed an almost linear behavior, or at least the sensor's resistance was correlated with the concentration. It should be taken into account that the concentration of the CXCL-13 gas that hits the sensor is much less than 1 pg / ml, considering the low evaporation rate of proteins. If this test is repeated in a vacuum chamber where the molecules can easily evaporate, the limit of detection of this sensor would be extensively improved. At a broader scale, the sensor underwent testing with spiked samples ranging from 1 pg / ml to 1 mg / ml. Notably, all biosensors possess inherent limitations in their detection range. However, this particular sensor exhibits an impressive detection range of 10A9 / ml, rendering it highly suitable for numerous applications beyond the capabilities of existing biosensors.

[0038] (0032) Selectivity: The same sensor was also tested against other similar cytokines. The main sensor response to other cytokines was a minor decrease in the resistance. Among the cytokines, CXCL-10 is most similar in structure to CXCL-13 and yet it was not detected, which shows an excellent selectivity of this sensor.

[0039] (0033) The graphs of Figures 2A, 2B, and 2C show the sensitivity test of the CXCL-13 gas sensor in a wide range of 1 pg / ml to 1 mg / m, selectivity test of the same sensor against other similar cytokines.

[0040] (0034) Comparison with ELISA: The diagrams and graphs of Figures 3A and 3B shows the comparison of the sensor reading with ELISA. Figure 3A is a diagram showing an enzyme-linked immunosorbent assay (ELISA). Figure 3B is a graph showing a comparison of the template molecule (CXCL-13) sensor and an enzyme-linked immunosorbent assay (ELISA). Owing to the processing time requirements of the utilized ELISA method, the entire ELISA procedure spanned two days to complete. In contrast, each measurement conducted by the sensor was remarkably swift, taking a mere 10 seconds. Despite the disparity in timeframes, the sensor readings were found to be in excellent agreement with the ELISA results with 85%. (0035) The diagrams and graphs of Figures 3A and 3B show a comparison of the sensor reading with enzyme-linked immunosorbent assay (ELISA). Although the sensor results were collected two days prior to ELISA, the sensor reading is still matching the ELISA result with 85%.

[0041] (0036) Application - Immobilization of the MIP sensor on a flexible, stretchable, and self healing substrate:

[0042] (0037) In a sensor embodiment that might be of value for wearable sensors, the same MIP sensor fabrication procedure was carried out using an elastomeric alginate polymer gel as the substrate for immobilization instead of silicon as the substrate. The alginate gel was mixed with the same rGo / PB mixture and then polymerized with IL-6 cytokine. The sensor was stabilized for 20 seconds, exposed to the 1 mg / ml of IL-6 from the second 20 to 40, and stabilized for another 20 seconds — as shown in the graph of Figure 4. Figure 4 is a graph showing the testing results of the template molecule (IL-6 cytokine) sensor. By using a wearable self-healed sensor, human skin is mimicked to develop artificial tissue-like electronic devices. In order to mimic human skin, a sensor needs to be stretchable and self-healed (be able to self recover after a cut). These skin-like electronic devices would be able to attach to the skin and directly detect biomarkers from breath, sweat, etc. These sensors are also capable of being connected to smart phones / watches and showing the testing results in real time.

[0043] (0038) The graph of Figure 4 shows a flexible sensor that detects IL-6 cytokine. The sensor reaction to the cytokine is evaluated by the resistance change. Only when the sensor is exposed to the positive sample (containing the specific template) does the resistance increase. Based on the graph, the sensor was able to detect IL-6 from an Eppendurf tube containing IL-6 as the resistance was increased from ~90KOhms to ~115KOhms.

[0044] (0039) Variations and Optional Features a. Conductive polymers other than polypyrrole (e.g., aniline, nordihydroguaiaretic acid) may be utilized to prepare the electrodes including combination of polymers (co-polymerisation). b. Integration of dissolving polymers may also be utilized to increase the porosity of the MIPs (e.g., polymers that dissolve in water for example salt crystals); degradation removal (e.g., proteins or protein aggregates); temperature dependent removal (e.g., poly(N -isopropyl acrylamide) (PNIPAAm)). c. Different transduction methods may be used as a mode of detection other than electrochemical sensing, including surface plasmon resonance, localized surface plasmon resonance, and others. d. Various nanomaterials may be utilized for the preparation of the nanocomposite, such as (carbon fibers, carbon nanotubes, metal nanoparticles, MXene). e. Different redox-active substances may be used for the preparation of nanocomposites (e.g., poly luminol, ferrocene, methylene blue). f. Although electrochemical polymerization has been used to prepare the sensors, other techniques such as electrospinning in-situ polymerization may be utilized. g. Polymerization techniques such as UV-cross linking auto-polymerization may be utilized based on the polymer of choice. h. Other stretchable polymers (e.g., poly dimethylsiloxane, SEBS, polyurethanes, etc.) may be used to prepare the sensors for wearable sensing.

[0045] (0040) Additional Advantageous Features:

[0046] (0041) As innovatively provided herein, the emulsion of rGo / PB and pyrrole is used to develop MIPs. And while Gr / PB and polypyrrole may have been used previously to fabricate sensors as separate layers, an innovation of the sensor provided mixes a conductive polymer with graphene, which has excellent electron mobility. Thus, instead of two separate layers where interference between the layers acts as the sensing mechanism such as that used in past MIPs, the innovations described (for example the described advantageous MIPs) provide graphene dispersed uniformly in the conductive polymer matrix. This gives the sensor the ability to use its whole volume as a sensing platform and not just the surface. Additionally, adjacency of graphene and pyrrole form a p-p isotype that builds the unique and innovative 3D arrangement of graphene resulting in the Fermi energy effect. (0042) The innovative self assembled and MIP formulation described also has important commercial value. Because the adhesion of graphene to a silicon substrate is poor, the graphene layer will often detach from the silicon surface once placed inside the electrolyte solution. However, with this method, a combination of graphene and the conductive polymer will be deposited directly on a silicon substrate. Notably, the methods provided herein help with consistency, fabrication time, and therefore may be more useful for large-scale production. The same concept was applied to a stretchable material — alginate — for the first time. The sensor was then successfully tested against the specific template molecule demonstrating the breadth of applications could include wearable materials or flexible sensors for industrial applications (e.g., on valves). Due to the high possibility of losing the sensors during the electrochemical sensing procedure, deposition of rGo / PB needed one extra day to dry on the silicon surface.

[0047] (0043) Specifically, in operation the present application provides and the sensing device includes:

[0048] • A device for sensing comprising: a supporting layer; and a sensing layer comprised of molecularly imprinted polymer (MIP) on the supporting layer, the MIP having a 3D matrix of graphene assembled from one or more layers of nanotextured reduced graphene with NH2, NH3, OH, O, and COOH, functionally forming a 3D matrix of graphene dispersed within the MIP, wherein the one or more layers of reduced nanotextured graphene forms a p-p isotype with the molecularly imprinted polymer. o The structure above wherein the graphene component is made of of aminated nanotextured reduced graphene with NH2 or NH3 functionalities of higher catalytic activities. o The structure above wherein the supporting layer is a silicon layer having an etched surface and the sensing layer is on the etched surface. The structure above wherein the 3D matrix of graphene comprises graphene and potassium ferrocyanide. The structure above wherein the MIP monomer is pyrrole or aniline or a conductive polymer. The structure above wherein the functional monomer is methacrylic acid with methacrylamide or dopamine, or a salt thereof, or a combination of any of the foregoing. The structure above wherein the graphene component has a thickness from 1 nm to 10 microns. The structure above wherein the graphene component is composed of nanoparticles, nanofibers, nanotriangles, or any nanodimension of reduced aminated graphene. The structure above wherein the graphene component is composed of nanotextured reduced aminated graphene with any dimension of nanotexturing including nanobumps, nanogrooves, nanopores, nanoedges, and nanoindents. The structure above wherein the graphene component has a surface roughness from 1 nm to 1 micron. The structure above wherein the graphene component is made of reduced aminated graphene with pore sizes from 1 nm to 1 micron. The structure above wherein the graphene component is made of reduced aminated graphene with a percent porosity from 1 - 99%. o The structure above wherein the sensing layer has a thickness in the range of 1 nanometer to 1 micron. o The structure above wherein the polymer is polypyrrole or a conductive polymer. o The structure above wherein the graphene component is made of of pentaminated nanotextured reduced graphene. o The structure above wherein the device is used to detect bacteria, viruses, cytokines, or any molecule useful for detecting and diagnosing diseases such as cancer, infection, inflammation, and neurological diseases. o The structure above wherein the graphene is uniformly dispersed within the MIP. o The structure above further comprising one or more electrical contacts. o The structure above wherein the support layer is made of silicon. o The structure above wherein the support layer is made of alginate.

[0049] • A device for sensing comprising: a support layer; and, a sensing layer on said support layer, said sensing layer having a three dimensional dispersion of aminated graphene in conductive polymer matrix, said matrix with cavities on the surface formed by self-assembled chemistry and which are tailored binding sites to a template molecule, said cavities surrounded by dopamine and aminated graphene. o The structure above further comprising one or more electrical contacts. The structure above wherein said cavities are formed by self-assembled chemistry of the template molecule and dopamine on the three dimensional matrix of graphene and conductive polymer matrix such as a polypyrrole matrix. The structure above wherein the graphene component is made of reduced aminated nanotextured graphene with NH2 and NH3 functionalities of higher conductivity. The structure above wherein the support layer is made of silicon having an etched surface and the sensing layer is on the etched surface. The structure above wherein the three dimensional dispersion of aminated graphene in the conductive polymer matrix such as a polypyrrole matrix comprises graphene and potassium ferrocyanide. The structure above wherein the aminated graphene is pentaminated graphene. The structure above wherein the functional monomer is methacrylic acid with methacrylamide or dopamine, or a salt thereof, or a combination of any of the foregoing. The structure above wherein the graphene component is from 1 nm to 10 microns. The structure above wherein the graphene component is composed of nanoparticles, nanofibers, nanotriangles, or any nanodimension of reduced aminated graphene. The structure above wherein the graphene component is composed of nanotextured reduced aminated graphene with any dimension of nanotexturing including nanobumps, nanogrooves, nanopores, nanoedges, and nanoindents. The structure above wherein the graphene component has a surface roughness from 1 nm to 1 micron. The structure above wherein the graphene component is made of reduced aminated graphene with pore sizes from 1 nm to 1 micron. The structure above wherein the graphene component is made of reduced aminated graphene with a percent porosity from 1 - 99%. The structure above wherein the sensing layer has a thickness in the range of 1 nanometer to 1 micron. The structure above wherein the conductive polymer is polypyrrole. The structure above wherein the device is used to detect bacteria, viruses, cytokines, or any molecule useful for detecting and diagnosing diseases such as cancer, infection, inflammation, and neurological diseases. The structure above wherein said sensing layer has a three dimensional uniform dispersion of aminated graphene in conductive polymer matrix such as a polypyrrole matrix. The structure above wherein the support layer is made of silicon. The structure above wherein the support layer is made of alginate. (0044) The foregoing description of the exemplary embodiments is provided to enable any person skilled in the art to make or use the claimed subject matter. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without the use of the innovative faculty. Thus, the claimed subject matter is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

CLAIMSWhat is Claimed is:

1. A device for sensing comprising: a supporting layer; and a sensing layer comprised of molecularly imprinted polymer (MIP) on said supporting layer, said MIP having a 3D matrix of graphene assembled from one or more layers of nanotextured reduced graphene with NH2, NH3, OH, O, and COOH, functionally forming a 3D matrix of graphene dispersed within said MIP, wherein the one or more layers of reduced nanotextured graphene forms a p-p isotype with the molecularly imprinted polymer (MIP).

2. The device in Claim 1, wherein the graphene component is made of of aminated nanotextured reduced graphene with NH2 or NH3 functionalities of higher catalytic activities.

3. The device of claim 1, wherein the supporting layer is a silicon layer having an etched surface and the sensing layer is on the etched surface.

4. The device of claims 1 or 2, wherein the 3D matrix of graphene comprises graphene and potassium ferrocyanide.

5. The device of claims 1-4, wherein the MIP monomer is pyrrole or aniline or a conductive polymer.

6. The device of claims 1-5, wherein the functional monomer is methacrylic acid with methacrylamide or dopamine, or a salt thereof, or a combination of any of the foregoing.

7. The device in Claim 1, wherein the graphene component has a thickness from 1 nm to 10 microns.

8. The device in Claim 1, wherein the graphene component is composed of nanoparticles, nanofibers, nanotriangles, or any nanodimension of reduced aminated graphene.

9. The device in Claim 1, wherein the graphene component is composed of nanotextured reduced aminated graphene with any dimension of nanotexturing including nanobumps, nanogrooves, nanopores, nanoedges, and nanoindents.

10. The device in Claim 1, wherein the graphene component has a surface roughness from 1 nm to 1 micron.

11. The device in Claim 1, wherein the graphene component is made of reduced aminated graphene with pore sizes from 1 nm to 1 micron.

12. The device in Claim 1, wherein the graphene component is made of reduced aminated graphene with a percent porosity from 1 - 99%.

13. The device of Claim 1, wherein the sensing layer has a thickness in the range of 1 nanometer to 1 micron.

14. The device of Claim 1, wherein the polymer is polypyrrole or a conductive polymer.

15. The device in Claim 1, wherein the graphene component is made of of pentaminated nanotextured reduced graphene.

16. The device of claim 1, wherein the device is used to detect bacteria, viruses, cytokines, or any molecule useful for detecting and diagnosing diseases such as cancer, infection, inflammation, and neurological diseases.

17. The device of claim 1, wherein said graphene is uniformly dispersed within said MIP.

18. The device of claim 1, further comprising one or more electrical contacts.

19. The device of claim 1, wherein the support layer is made of silicon.

20. The device of claim 1, wherein the support layer is made of alginate.

21. A device for sensing comprising: a support layer; and,a sensing layer on said support layer, said sensing layer having a three dimensional dispersion of aminated graphene in conductive polymer matrix, said matrix with cavities on the surface formed by self-assembled chemistry and which are tailored binding sites to a template molecule, said cavities surrounded by dopamine and aminated graphene.

22. The devices of claim 21, further comprising one or more electrical contacts.

23. The devices of claim 21, wherein said cavities are formed by self-assembled chemistry of the template molecule and dopamine on the three dimensional matrix of graphene and conductive polymer.

24. The device in Claim 21, wherein the graphene component is made of reduced aminated nanotextured graphene with NH2 and NH3 functionalities of higher conductivity.

25. The device of claim 21, wherein the support layer is made of silicon having an etched surface and the sensing layer is on the etched surface.

26. The device of claims 21 or 24, wherein the three dimensional dispersion of aminated graphene in conductive polymer matrix comprises graphene and potassium ferrocyanide.

27. The device of any one of claims 21-26, wherein the aminated graphene is pentaminated graphene.

28. The device of any one of claims 21-27, wherein the functional monomer is methacrylic acid with methacrylamide or dopamine, or a salt thereof, or a combination of any of the foregoing.

29. The device in Claim 21, wherein the graphene component is from 1 nm to 10 microns.

30. The device in Claim 21, wherein the graphene component is composed of nanoparticles, nanofibers, nanotriangles, or any nanodimension of reduced aminated graphene.

31. The device in Claim 21, wherein the graphene component is composed of nanotextured reduced aminated graphene with any dimension of nanotexturing including nanobumps, nanogrooves, nanopores, nanoedges, and nanoindents.

32. The device in Claim 21, wherein the graphene component has a surface roughness from 1 nm to 1 micron.

33. The device in Claim 21, wherein the graphene component is made of reduced aminated graphene with pore sizes from 1 nm to 1 micron.

34. The device in Claim 21, wherein the graphene component is made of reduced aminated graphene with a percent porosity from 1 - 99%.

35. The device of Claim 21, wherein the sensing layer has a thickness in the range of 1 nanometer to 1 micron.

36. The device of Claim 21, wherein the conductive polymer is polypyrrole.

37. The device of claim 21, wherein the device is used to detect bacteria, viruses, cytokines, or any molecule useful for detecting and diagnosing diseases such as cancer, infection, inflammation, and neurological diseases.

38. The device of claim 21, wherein said sensing layer has a three dimensional uniform dispersion of aminated graphene in the conductive polymer matrix.

39. The device of claim 21, wherein the support layer is made of silicon.

40. The device of claim 21, wherein the support layer is made of alginate.

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