WO2019173220A1 - Système et procédé de détection sans fil d'état de santé buccale - Google Patents

Système et procédé de détection sans fil d'état de santé buccale Download PDF

Info

Publication number
WO2019173220A1
WO2019173220A1 PCT/US2019/020559 US2019020559W WO2019173220A1 WO 2019173220 A1 WO2019173220 A1 WO 2019173220A1 US 2019020559 W US2019020559 W US 2019020559W WO 2019173220 A1 WO2019173220 A1 WO 2019173220A1
Authority
WO
WIPO (PCT)
Prior art keywords
cancer
acid sequence
amino acid
binding
electrical signal
Prior art date
Application number
PCT/US2019/020559
Other languages
English (en)
Inventor
Marcelo FREIRE
Original Assignee
J. Craig Venter Institute, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by J. Craig Venter Institute, Inc. filed Critical J. Craig Venter Institute, Inc.
Publication of WO2019173220A1 publication Critical patent/WO2019173220A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/14507—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue specially adapted for measuring characteristics of body fluids other than blood
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/1486—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using enzyme electrodes, e.g. with immobilised oxidase
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
    • A61B5/6801—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be attached to or worn on the body surface
    • A61B5/6813—Specially adapted to be attached to a specific body part
    • A61B5/6814—Head
    • A61B5/682—Mouth, e.g., oral cavity; tongue; Lips; Teeth
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B2562/00—Details of sensors; Constructional details of sensor housings or probes; Accessories for sensors
    • A61B2562/02—Details of sensors specially adapted for in-vivo measurements
    • A61B2562/0271—Thermal or temperature sensors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/14532—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue for measuring glucose, e.g. by tissue impedance measurement
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/1495—Calibrating or testing of in-vivo probes

Definitions

  • the present disclosure relates generally to the field of devices having biosensors, and particularly to devices having biosensors configured to detect, monitor and track health information of a subject, such as a human or animal, as presented by biomarkers available in biofluids, such as saliva and gingival crevicular fluid (GCF).
  • a subject such as a human or animal
  • biomarkers available in biofluids such as saliva and gingival crevicular fluid (GCF).
  • biofluid monitoring a device bonded to the tooth surface with continuous and in vivo oral biofluid monitoring has not yet been developed and/or publicly implemented. Inter individual differences are not captured in studies with average data. Democratization of such biofluid monitoring can facilitate personalized medicine, diagnostics, improve the molecular understanding of therapies and treatment protocols, and can improve health monitoring in the population. Accordingly, the need for improved diagnostic devices and systems for identifying biomarkers present in biofluids is manifest.
  • Mammalian saliva carries a precise developmental and biological program that defines the function of multiple organs including oral and periodontal tissues (tissues surrounding the tooth). Saliva is at the interface of dynamic interactions between microbial ecosystems and oral immune cells.
  • Elevated glucose or insulin in saliva for example, activates dysbiosis (disease) of tissues, accelerating systemic inflammation, leading to tissue loss and loss of function.
  • Saliva provides an optimal buffering system for microbial acidic metabolites maintaining microbial community homeostasis in a neutral pH and a constant supply of nutrients to the environment ( see Scannapieco, F. A. (1994), Saliva-bacterium interactions in oral microbial ecology. Critical Review s in Oral Biology & Medicine, 5(3), 203- 248).
  • microorganisms are sensitive to minor changes in the local environment, including pH, nutrients, oxygen, moisture, and host immune responses. Changes in saliva flow rate (i.e. in dry mouth patient) or pH (i.e.
  • salivary components are known to affect bacterial growth and lipopolysaccharide (LPS) production, e.g. pH modulating molecules such as bicarbonates, phosphates, and urea, salivary proteins contribute to processes associated with microbial metabolism, aggregation, attachment of biofilms ( see Kolenbrander, P. E.
  • Human salivary cells are comprised of white blood cells, epithelial cells. As the most abundant leukocytes in saliva, it is estimated that 30,000 polymorphonuclear neutrophils (PMNs) cells (neutrophils) transit through periodontal tissue every minute (see Tonetti, M. S., Imboden, M. A., & Lang, N. P. (1998). Neutrophil migration into the gingival sulcus is associated with transepithelial gradients of interleukin-8 and ICAM-l, Journal of periodontology, 69(10), 1139-1147). These cells can be detected in saliva, through their receptor ERV1 receptor.
  • PMNs polymorphonuclear neutrophils
  • PMNs are important source of inflammatory mediators of the host immune response against proliferating pathogenic microorganisms.
  • Inflammatory cytokines such as c-reactive protein (CRP), Interleukin (IL)-6, 1-1 and others can be monitored through saliva instead of the blood.
  • PMNs could form a barrier between the junctional epithelium and the subgingival biofilm (at the gingival crevice), preventing the apical migration of the bacteria, which is essential for control of bacterial invasion.
  • salivary PMNs are often adapted in evaluating oral and systemic health and treatment outcomes.
  • Salivary protein content is vast with total of 5700 quantified proteins, including
  • a device designed to attach to the tooth surface protect the internal sensing device from the oral external disturbances to sense salivary biological and molecular information.
  • Graphene based oral device or system is inserted into a subject’s body to continuously monitor biological changes. This innovation is useful for real-time data collection, live monitoring and improved patient care (e.g. diagnosis and monitoring patient response to therapies).
  • the present disclosure provides a method of binding one or more analytes from the biological fluid, comprising attaching a binding moiety to an electrically conductive surface, wherein said binding moiety interacts specifically with one or more biomarkers in the biological fluid, and wherein upon binding a detectable signal is generated, which corresponds to an amount and/or presence of the biomarker in a biological fluid such as saliva and GCF.
  • said electrically conductive surface is a graphene or graphene oxide-based material, which may be provided on a non-biodegradable support such as a non-biodegradable polymer, titanium, metal, composite resin, plastic or ceramic.
  • the non-biodegradable support may preferably be configured for placement in the gingival sulcus or periodontal pocket of the users’ oral cavity but placement can be in other locations in or on the body and said biological fluid can be saliva, crevicular fluid, lymphatic fluid, spinal fluid, or oral sweat.
  • the device is placed in the supra or sub-gingival space, preferably proximal to the cementoenamel-junction (CEJ) (for instance coronal to the cementoenamel-junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth). Accordingly, the placement of the device can provide molecular information from the supra and sub-gingival space.
  • CEJ cementoenamel-junction
  • the biomarker comprises one or more of an inflammatory molecule, a cytokine, a bacterial metabolite, a bacterial membrane component, a bacterial cell wall component, a host cell, DNA strands from host cells, immune cell wall, a virus, a nonviral pathogen, a polysaccharide, a lipid, or a nucleic acid, or any combination thereof, and in preferred embodiments, the biomarker comprises one or more of c-reactive protein (CRP), lipopolysaccharide (LPS), glucose, lactate, ERV-l (or chemR23, CMKLR1), or cortisol, or any combination thereof.
  • CRP c-reactive protein
  • LPS lipopolysaccharide
  • glucose lactate
  • ERV-l or chemR23, CMKLR1
  • cortisol cortisol
  • the methods of the present disclosure may further comprise transmitting information, preferably by wireless transmission, regarding the presence and/or concentration of said biomarkers to an external device and, preferably may be carried out in the oral cavity, especially from the oral mucosa, and in the periodontal space or in gingival sulcus.
  • the methods are carried out in the supra or sub-gingival space, preferably proximal to the cementoenamel-junction (CEJ) (for instance coronal to the cementoenamel-junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth).
  • CEJ cementoenamel-junction
  • the present disclosure further contemplates a system or device for detecting the presence of one or more biomarkers in a biological fluid
  • a biological fluid comprising an electrically conductive layer, which may be provided on a non-biodegradable support such as a non-biodegradable polymer, plastic, titanium, composite resin or ceramic and, which may preferably be configured for placement on the tooth surface at the gingival crevice or at the base of the periodontal pocket of a subject’s oral cavity and, which has attached thereto one or more binding moieties configured for the binding of one or more biomarkers, wherein binding of one or more biomarkers to said binding moieties generates a change in one or more electronic characteristics of said conductive layer; and further comprising one or more base layers in contact with said conductive surface, and an antenna, whereby said change in one or more electronic characteristics related to the binding of said one or more biomarkers may be communicated, preferably wirelessly, to an external device.
  • a non-biodegradable support such as a non-bio
  • the device is placed in the supra or sub-gingival space, preferably proximal to the cementoenamel- junction (CEJ) (for instance coronal to the cementoenamel-junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth).
  • CEJ cementoenamel- junction
  • said electrically conductive surface is a graphene or graphene oxide and said biological fluid may be saliva, crevicular fluid, lymphatic fluid, spinal fluid or oral sweat.
  • the biomarkers contemplated herein may comprise one or more of an inflammatory molecule, a cytokine, a bacterial metabolite, a bacterial membrane component, a bacterial cell wall component, a virus, a nonviral pathogen, a polysaccharide, a lipid or a nucleic acid, or any combination thereof, especially (1) derived from inflammation, c-reactive protein (CRP), Erv-l; (2) derived from the microbiome, bpopolysaccharide (LPS); (3) derived from metabolism glucose, lactate, or cortisol, or any combination thereof.
  • an inflammatory molecule especially (1) derived from inflammation, c-reactive protein (CRP), Erv-l; (2) derived from the microbiome, bpopolysaccharide (LPS); (3) derived from metabolism glucose, lactate, or cortisol, or any combination thereof.
  • the systems and devices of the present disclosure may further comprise a transmitter, which transmits information concerning the presence and/or concentration of said biomarkers to an external device, preferably wirelessly, and may be deployed within the oral cavity, especially within the gingival crevice or periodontal space or on other parts of the body e.g., in contact with the skin.
  • the device is placed in the supra or sub-gingival space, preferably proximal to the cementoenamel -junction (CEJ) (for instance coronal to the cementoenamel- junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth).
  • CEJ cementoenamel -junction
  • the systems and devices as described herein may further comprise an external device in the shape of a custom orthodontic device configured to receive a signal from said change in one or more electronic characteristics of said conductive layer, wherein said external device may comprise a processor and a memory configured to retain and/or analyze said signal from said change in one or more electronic characteristic of said conductive layer.
  • said system or device, as described herein further comprises a base layer, said base layer may incorporate one or more of a ceramic, composite resin, titanium, metal oxide, or a polymer layer.
  • a method of binding one or more analytes in a biological fluid from a subject comprising attaching a binding moiety to an electrically conductive surface, wherein said electrically conductive surface is joined to a support, such as a support comprising ceramic, silica, or a plastic or a polymer, wherein said support is, optionally configured for introduction into the oral cavity of said subject, preferably in the gingival crevice or periodontal space of said subject, wherein said binding moiety interacts specifically with one or more biomarkers, and, wherein binding of said binding moiety and said one or more biomarkers generates a detectable signal corresponding to the amount or presence of said biomarker in the biological fluid of said subject.
  • said electrically conductive surface is a graphene or a graphene oxide.
  • biomarker comprises one or more of an inflammatory molecule, a cytokine, a bacterial metabolite, a bacterial membrane component, a bacterial cell wall component, a virus, a nonviral pathogen, a polysaccharide, a lipid, or a nucleic acid, or any combination thereof.
  • biomarker comprises one or more of c-reactive protein, bacterial lipopolysaccharide, glucose, lactate, insulin, or cortisol, or any combination thereof.
  • a device for detecting the presence of one or more biomarkers in a biological fluid from a subject comprising:
  • a housing structure configured to prevent a flow of matter from an exterior of the housing structure to an interior of the housing structure, the housing structure comprising a base and a cover, wherein the base forms a cavity and comprises an aperture that provides a path from the exterior of the housing structure to the cavity, and, wherein the housing structure is configured to be exposed to the biological fluid;
  • a support structure comprising a sensor portion and a coupling portion, the sensor portion configured to extend through the aperture, wherein the coupling portion and the sensor portion prevent the flow of matter from the exterior of the housing structure into the cavity when the sensor portion is extended through the aperture;
  • an electrically conductive layer optionally comprising a binding moiety, disposed on the sensor portion of the support structure and configured to convey an electrical signal based on exposure of the sensor portion to the one or more biomarkers;
  • control circuit conductively coupled to the electrically conductive layer and configured to: detect the electrical signal
  • the electrically conductive layer has attached thereto one or more binding moieties, preferably one of a protein, peptide, antibody, aptamer, or binding fragment thereof, such as an scFv configured for the binding of the one or more molecular markers, wherein the binding of the one or more biomarkers to said binding moieties generates a change in one or more electronic characteristics of the electrically conductive layer and, wherein the electrical signal is a result of the generated change in the one or more electrical characteristics.
  • binding moieties preferably one of a protein, peptide, antibody, aptamer, or binding fragment thereof, such as an scFv configured for the binding of the one or more molecular markers, wherein the binding of the one or more biomarkers to said binding moieties generates a change in one or more electronic characteristics of the electrically conductive layer and, wherein the electrical signal is a result of the generated change in the one or more electrical characteristics.
  • the support structure comprises a plurality of additional sensor structures, each of the plurality of additional sensor portions having disposed thereon an electrically conductive layer configured to convey an electrical signal based on exposure of the respective additional sensor portion to the one or more biomarkers
  • the base comprises a plurality of additional apertures through which one of the plurality of additional sensor portions is configured to extend, and wherein the electrically conductive layer disposed on at least one of the plurality of additional sensor portions has attached thereto a different one or more binding moieties from the one or more binding moieties attached to the electrically conductive layer disposed on the sensor portion.
  • control circuit comprises one or more of a hardware processor configured to analyze the electrical signal, a memory configured to store the information derived from the electrical signal, and a transmission circuit configured to transmit the electrical signal or information derived therefrom.
  • the one or more biomarkers comprise one or more of an inflammatory molecule, a cytokine, a bacterial metabolite, a bacterial membrane component, a bacterial cell wall component, a virus, a nonviral pathogen, a polysaccharide, a lipid, or a nucleic acid, or any combination thereof.
  • biomarkers comprises one or more of c-reactive protein (CRP), bacterial lipopolysaccharide (LPS), glucose, lacatate, insulin, Erv-l, or cortisol, or any combination thereof.
  • CRP c-reactive protein
  • LPS bacterial lipopolysaccharide
  • control circuit is further configured to prepare the electrical signal or information derived therefrom for transmission to an external device configured to receive a wired or wireless transmission from the device.
  • control circuit is further configured to prepare the electrical signal or information derived therefrom for transmission to an external device configured to retain or analyze the electrical signal.
  • one or more of the housing structure and the support structure comprises a non- conductive material, preferably one or more of ceramic, silica, a polymer, polyethylene, metal, titanium, composite resin or plastic.
  • an additional change in one or more electronic characteristics of the electrically conductive layer disposed on the one of the plurality of additional sensor portions based on binding one of the one or more biomarkers to one or more binding moieties, preferably one of a protein, peptide, antibody, aptamer, or binding fragment thereof, such as an scFv attached to the electrically conductive layer of the one of the plurality of additional sensor portion; and conveying another electrical signal, via the electrically conductive layer disposed on the one of the plurality of additional sensor portions, to the control circuit,
  • the electrically conductive layer disposed on the one of the plurality of additional sensor portions has attached thereto a different one or more binding moieties from the one or more binding moieties attached to the electrically conductive layer disposed on the sensor portion and wherein the other electrical signal is a result of the generated change in the one or more electrical characteristics of the electrically conductive layer disposed on the one of the plurality of additional sensor portions.
  • biomarkers comprise one or more of an inflammatory molecule, a cytokine, a bacterial metabolite, a bacterial membrane component, a bacterial cell wall component, a virus, a nonviral pathogen, a polysaccharide, a lipid or a nucleic acid, or any combination thereof.
  • biomarkers comprises one or more of c-reactive protein, bacterial lipopolysaccharide, glucose, lactate, insulin, Erv-l or cortisol, or any combination thereof.
  • one or more of the housing structure and the support structure comprises a non-conductive material, preferably one or more of ceramic, silica, polyethylene, titanium, composite resin, a polymer, metal, or plastic.
  • a method of binding an analyte or biomarker comprising using a device of any of alternatives 9-27.
  • the device comprises an antibody or binding fragment thereof, such as an scFv which is specific for c-reactive protein, bacterial lipopoly saccharide, glucose, lactate, insulin, Erv-l or cortisol, or any combination thereof and the device is configured for placement within the periodontal space or gingival crevice of a subject or in or near the supra or sub-gingival space, preferably proximal to the cementoenamel-j unction (CEJ) or coronal to the cementoenamel-junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth.
  • an antibody or binding fragment thereof such as an scFv which is specific for c-reactive protein, bacterial lipopoly saccharide, glucose, lactate, insulin, Erv-l or cortisol, or any combination thereof and the device is configured for placement within the periodontal space or gingival crevice of a subject or
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, and/or 16, or a biologically active variant or binding fragment thereof, or any combination thereof.
  • scFv comprising an amino acid sequence that comprises at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, and/or 16, or a biologically active variant or binding fragment thereof, or any combination thereof.
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises a variable heavy chain (VH) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 6, 10, and 14 or a biologically active variant or binding fragment thereof, or any combination thereof.
  • VH variable heavy chain
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises a variable heavy chain (VH) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 6, 10, and 14 or a biologically active variant or binding fragment thereof, or any combination thereof.
  • VH variable heavy chain
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises a variable light chain (VK) sequence comprising an amino acid sequence that comprises at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 4, 8, 12, and 16, or a biologically active variant or binding fragment thereof, or any combination thereof.
  • VK variable light chain
  • [0071] 54 The device of anyone of Alternatives 9-26, wherein the binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises a variable heavy chain (VH) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 2 and a variable light chain (VK) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 4.
  • VH variable heavy chain
  • VK variable light chain
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises a variable heavy chain (VH) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 6 and a variable light chain (VK) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 8.
  • VH variable heavy chain
  • VK variable light chain
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises a variable heavy chain (VH) sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 10 and a variable light chain (VK) sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the ammo acid sequence to the amino acid sequence set forth in SEQ ID NO: 12.
  • VH variable heavy chain
  • VK variable light chain
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, comprising an amino acid sequence that comprises a variable heavy chain (VH) sequence comprising an amino acid that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 14 and a variable light chain (VK) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 16.
  • VH variable heavy chain
  • VK variable light chain
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, designated by, derived from, or obtainable fromlGl2.G8-2.C3, 5G4.F8-1.G5, 5G4.F8-1B7, 5G4.F8-1.C10, clone ab65333, ab6533 l, clone 5A9, 2D7/1, or clone ab2000l l .
  • a biomarker for identifying the presence or amount or concentration of a biomarker in a subject, such as a human or animal, for example a cow, a pig, a sheep, a horse, a dog, or a cat.
  • binding moiety is an antibody or binding fragment thereof, such as an scFv, designated by, derived from, or obtainable from 1G12.G8-2.C3, 5G4.F8-1.G5, 5G4.F8-1B7, 5G4.F8-1.C10, clone ab65333, ab6533 l, clone 5A9, 2D7/1, or clone ab2000l l.
  • an scFv designated by, derived from, or obtainable from 1G12.G8-2.C3, 5G4.F8-1.G5, 5G4.F8-1B7, 5G4.F8-1.C10, clone ab65333, ab6533 l, clone 5A9, 2D7/1, or clone ab2000l l.
  • 48, 50, 52, 54, 56, 58, 60, or 62 for identifying the presence or amount or concentration of a biomarker in a subject, such as a human or animal, for example a cow, a pig, a sheep, a horse, a dog, or a cat comprising contacting the device of anyone of Alternatives 9-26 or Alternatives 48, 50, 52, 54, 56, 58, 60, or 62 with said subject and determining the presence or concentration of said biomarker from the signal provided by the device.
  • biomarker is an inflammatory molecule, a cytokine, a bacterial metabolite, a bacterial membrane component, a bacterial cell wall component, a virus, a nonviral pathogen, a polysaccharide, a lipid or a nucleic acid, or any combination thereof, such as c-reactive protein, bacterial lipopolysaccharide, glucose, lactate, insulin, Erv-l or cortisol, or any combination thereof.
  • a disease or condition associated with a disease or for monitoring the progression of a disease or a therapy for a disease in a subject for example a cow, a pig, a sheep, a horse, a dog, or a cat.
  • 48, 50, 52, 54, 56, 58, 60, or 62 for identifying the presence of a disease or condition associated with a disease or for monitoring the progression of a disease or a therapy for a disease in a subject, for example a cow, a pig, a sheep, a horse, a dog, or a cat comprising contacting the device of anyone of Alternatives 9-26 or Alternatives 48, 50, 52, 54, 56, 58, 60, or 62 with said subject and determining the presence or concentration of said biomarker from the signal provided by the device.
  • Figure 1 shows schematic representations of one embodiment of the systems and methods of the present disclosure, according to an exemplary embodiment.
  • Figure 2 shows a breakdown of information available from an analysis of saliva biofluid, in accordance with an exemplary embodiment.
  • Figure 3A shows a perspective view of a device used to input and analyze biofluids, in accordance with an exemplary embodiment.
  • Figure 3B shows a front view of the device of Figure 3 A, in accordance with an exemplary embodiment.
  • Figure 4 A shows a front schematic view of the cover of the device of Figure
  • Figure 4B shows a side perspective view of the cover of Figure 4A, in accordance with an exemplary embodiment.
  • Figure 4C shows a series of three cross-section views of the cover of Figure 4A, detailing how a gasket in the groove adapts as the cover of Figure 4A is attached to the device of Figure 3 A, in accordance with an exemplary embodiment.
  • Figure 5 shows a perspective view of the base housing of the device 300 of
  • FIG. 3 A in accordance with an exemplary embodiment.
  • Figures 6A and 6B are schematics showing exemplary dimensions of the base housing of Figure 5, in accordance with an exemplary embodiment.
  • Figures 7A-7C show different views of sensor housing components and sensor channels of the device of Figure 3 A, in accordance with an exemplary embodiment.
  • Figure 8 shows a perspective view of the base housing of Figure 5 combined with the sensor housing component of Figures 7A-7C.
  • Figures 9A and 9B show perspective views of a housing attachment that attaches the device of Figure 3A to a surface of a subject’s body, in accordance with an exemplary embodiment.
  • Figure 10 illustrates a block diagram of an aspect of a circuit coupled to the sensor channels of the device of Figure 3 A, in accordance with an exemplary embodiment.
  • Whole saliva is a composite biofluid providing information from crevicular fluids, serum, interstitial fluids, host cell secretions, microbial contents, water, and oral sweat.
  • saliva and blood have similar compositions.
  • various information can be obtained from saliva, highlighting the diagnostic potential of saliva to reveal physiological and pathological changes.
  • High-throughput screening techniques applied to saliva may provide identification and quantification of a host and microbial DNA, RNA, proteins, and metabolites. For example, elevated pro-inflammatory signals such as acute phase proteins produced by the liver-C-reactive protein (CRP) are shown to be elevated in the serum and saliva of uncontrolled diabetic individuals.
  • CRP liver-C-reactive protein
  • Saliva may include the body’s systemic molecular information. Saliva is an essential biofluid responsible for shaping oral and systemic health. When the body’s saliva physiology is in dysregulation, the systemic system may also be disturbed. At the molecular level, secretomes from microbes and human proteomes of saliva overlap with other fluids such as blood serum and gingival crevicular fluid (GCF), as evidenced by various studies of saliva with regards to providing systemic information. Mapping of overlapping proteins from plasma and saliva through high resolution mass spectrometry for protein identification has resulted in identification of 1,505 common proteins and 690 proteins that are shared among saliva, plasma, and gingival fluid.
  • GCF gingival crevicular fluid
  • Figure 1 shows schematic representations of one embodiment of the systems and methods of the present disclosure, according to an exemplary embodiment.
  • Figure 1 shows a derivatizable graphene layer, which is then printed through a process of printing graphene layers into a graphene layer structure (as described in more detail below).
  • Functionalization of one or more graphene layers may involve integrating one or more antibodies or binding fragments thereof e.g., scFvs, enzymes, nucleic acids, and/or peptides with the one or more graphene layers.
  • the integrated graphene layer(s) may then be used for in vitro or in vivo sensing.
  • the in vitro and/or in vivo sensing may involve the use of exemplary systems and methods for single analyte detection, multiplex analyte detection, and incorporation of the systems/methods into a biosensor, and transmission of data from the detection events to external devices for analysis, in accordance with an exemplary embodiment.
  • Figure 2 shows a breakdown of information available from an analysis of saliva biofluid, in accordance with an exemplary embodiment.
  • the breakdown shows that saliva biofluid provides an array of host-microbial molecular information.
  • Figure 2 also shows schematics depicting current technologies to assay salivary molecules (i.e., DNA, and 16S sequencing shotgun metagenomics sequencing (SMS), host mRNA or microbial mRNA expression), proteomics, comparative metabolomics analysis, and mass spectrometry (MS) based network infrastructures, e.g., the Global Natural Products Social (GNPS).
  • SMS DNA, and 16S sequencing shotgun metagenomics sequencing
  • MS mass spectrometry
  • an understanding that the host’s physiological landscape is shaped by dynamic molecular changes that are temporally regulated introduces a new need to develop novel devices, systems, methods, and apparatus for in vivo” (in body) monitoring.
  • the device described herein may be designed and manufactured for in vivo testing and monitoring. For example, aspects of the device described herein continuously monitor molecular information and detect multiplex changes in vivo, in real time.
  • the device is able to, when placed in an oral cavity, for example: (1) detect saliva biomarkers related to systemic health and a systemic condition; (2) monitor functional changes of the saliva to understand unknown host-microbial-metabolic patterns at the individual level; (3) assist in investigating, in real-time, predictors of phenotypic changes; (4) move beyond assessing only limited number of biomarkers for monitoring, and (5) address multiplex capacity for in vivo detection of saliva.
  • the device described herein may provide a framework for noninvasive monitoring through saliva molecular detection, provide for improved functional host-microbiome-metabolic studies, and enable the capture of host-microbial-metabolism interactions.
  • Exemplary benefits of utilizing or focusing on saliva in experimental and clinical assessment over other biofluids, such as blood, include the non-invasive collection of saliva, ease of access of the oral cavity, minimal risk to the donor, and low-cost processing, among others.
  • the device may be tailored to present high sensitivities for continuous molecular detection.
  • the device described herein may monitor a biofluid (for example, saliva or gingival fluid or oral sweat) that is readily accessible in a minimally invasive manner.
  • a biofluid for example, saliva or gingival fluid or oral sweat
  • FIG. 3A shows a perspective view of a device 300 used to input and analyze biofluids, in accordance with an exemplary embodiment.
  • the device 300 includes various components or parts.
  • the device 300 includes a cover 302 and a base housing 304.
  • the base housing includes a plurality of apertures 305.
  • the apertures 305 are positioned in one or more rows along one or more sides of the base housing 304.
  • the device 300 may be generally shaped to conform to a surface to which it is attached or coupled.
  • the device 300 when the device 300 is attached to a tooth of a host, the device 300 may be generally shaped to conform to the shape of the tooth and, thus, may be generally shaped like a tooth surface and anatomical features, protecting or not harming the adjacent structures.
  • Figure 3B shows a front view of the device 300 of Figure 3A, in accordance with an exemplary embodiment.
  • the device 300 includes a plurality of apertures 305 through which a plurality of sensor channels 306 extend.
  • the sensor channels 306 comprise or are formed from one or more graphene or graphene oxide layers, formed as described above though a graphene or graphene oxide printing process or similar methodology.
  • the entire structure of the sensor channels 306 are formed from printed or otherwise disposed graphene or graphene oxide layers.
  • the sensor channels 306 comprise a base material (for example, an insulating and/or medically safe material, such as a plastic, polymer, titanium, composite resin, or ceramic material) on which the one or more graphene or graphene oxide layers are disposed.
  • the graphene or graphene oxide layers are replaced with one or more other material layers having similar properties to the graphene or to the graphene oxide layers. Accordingly, at least an outer surface of the sensor channels 306 is generally low-cost, flexible, highly conductive, and water repellant.
  • the sensor channels 306 may extend through the one or more rows of apertures 305 along the one or more sides of the base housing 304.
  • the device 300 includes at least one row of apertures on each side of the device 300, where the sides of the device 300 are curved and, wherein the rows of apertures 305 and the sensor channels 306 that extended there between are generally aligned horizontally across the device 300 in rows.
  • Figure 4A shows a front schematic view of the cover 302 of the device 300 of
  • the front of the cover 302 includes one or more portions 402 that extend or protrude away from a surface 401 of the front of the cover 302 in horizontal and vertical directions across the cover 302.
  • the portion 402 begins protruding away from the surface 401 in the vertical direction at a first point (for example, approximately l/5 th or l/4 th down a height of the cover 302 from a top edge of the cover 302) and be tapered such that as the portion 402 extends down the height of the cover 302, a distance by which the portion 402 protrudes from the surface 401 increases until the portion 402 reaches a bottom edge of the cover 302.
  • the portion 402 also begins to protrude away from the surface
  • the portion 402 protrudes from the surface 401 increases until the portion 402 reaches the sides of the cover 302.
  • the portion 402 is generally shaped like a triangle or an exterior surface of the user’s tooth to which the device 300 is attached.
  • the cover 302 may be a simple cover that provides a flat or slightly rounded surface to protect adjacent tissues.
  • the cover 302 may be shaped to accommodate components within the device 300.
  • Figure 4B shows a side perspective view of the cover 302 of Figure 4A, in accordance with an exemplary embodiment.
  • the view shows that the cover 302 has a depth or lip portion that may be inserted into the base housing 304 of the device, along which is a groove or channel 403 that runs around a perimeter and/or all edges or sides of the cover 302.
  • the groove 403 may provide a location in which a gasket or similar component is disposed and, which provides a waterproof and/or airtight seal around the device 300 such that no liquids, gasses, or solids are able to penetrate the housing of the device 300, as described in more detail below with regard to Figure 4C.
  • Figure 4C shows a series of three cross-section views of the cover 302 of Figure
  • FIG. 4A detailing how a gasket 405 in the groove 403 adapts as the cover 302 of Figure 4A is attached to the device 300 of Figure 3A, in accordance with an exemplary embodiment.
  • the gasket 405 changes shape and position as the cover 302 and its lip or depth portion extends into the base housing 304.
  • the gasket 405 appears generally disposed in a static or slightly compressed position as the cover 302 is merely positioned against the base housing 304.
  • the gasket 405 is shown compressed under a pressure P in a horizontal direction from left to right as the lip portion of the cover 302 is pressed into the base housing 304 such that the cover 302 and the base housing 304 are coupled to each other in an intermediate position.
  • the gasket 405 is compressed and extends into a gap 407 between the cover 302 and the base housing 304, thereby providing a seal between the cover 302 and the base housing 304. Accordingly, the gasket 405 prevents any flow of liquids, gases, and/or solids to pass from outside the device 300 inside the device 300 when the cover 302 and the base housing 304 are coupled together with the gasket 405 between them.
  • Figure 5 shows a perspective view of the base housing 304 of the device 300 of
  • the base housing 304 includes at least one row of apertures 305 along the sides of the base housing 304. As shown in Figure 5, the base housing 304 includes two rows of apertures 305 along each vertical side of the base housing 304, where the rows of apertures along each side are substantially aligned in two columns and fifteen rows.
  • the base housing 304 may include a cavity or volume 502 that generally contains one or more structures that support and/or otherwise include the sensor channels 306 (for example, the sensor housing structures described in more detail below with reference to Figures 7A-7C).
  • the cavity 502 within the base housing 304 may also house an electronic circuit or other circuit components configured to sense a signal from the sensor channels 306 and process, store, and/or convey the signals and/or information determined based on the signals. Additional details regarding the electronic circuit and other components is provided below with reference to Figure 10.
  • the base housing 304 may be constructed from any material that is safe to be inserted into a body.
  • the base housing 304 may be constructed from plastic, polyethylene, a polymer, ceramic, titanium, composite resin, steel, graphene or graphene oxide.
  • the base housing 304 may not be constructed from graphene or graphene oxide but may include one or more layers of graphene or graphene oxide disposed on one or more surfaces of the base housing 304, for example, inside the cavity 502.
  • the base housing 304 may be formed from and/or covered by an insulating material. By making at least the surfaces of the base housing 304 insulating, the sensor channels 306 may remain electrically isolated from each other.
  • FIGS 6A and 6B are schematics showing exemplary dimensions of the base housing 304 of Figure 5, in accordance with an exemplary embodiment.
  • the displayed dimensions exemplify the potential compactness of the device 300, which includes the cavity 502 of the base housing 304, within which the sensor channel 306 and the circuit or components that process, store, and/or convey information and/or the signals received from the sensor channels 306 are disposed.
  • the cavity 502 has a height of 2.8mm, a width of l .8lmm, and a depth of 0.55mm while the base housing 304 has a height of 3. lmm, a width of 2.3mm, and a depth of 0.7mm.
  • Figures 6A and 6B also show that the apertures 305 are substantially square shaped with sides of length 0.1 mm. Therefore, the sensor channels 306 that extend through the apertures 305 (as shown in Figure 3B) are substantially square shaped with sides of just less than 0.1 mm. In some embodiments, the apertures 305 and the sensor channels 306 may be formed in any matching shape (circular, oval, elliptical, rectangular, polygonal, etc.) so long as the apertures 305 and the sensor channels 306 are the same shape.
  • the sensor channels 306 and the apertures 305 are sized such that when the sensor channels 306 are retracted, they form a seal with the apertures such that no matter flows from outside the device 300 into the device 300 (for example, into the cavity 502). Additionally, when the sensor channels 306 are extended out of the device 306, the sensor housing components 702 and the sensor channels 306 are configured to form a seal with the apertures 305 and the cavity 502 to prevent matter from flowing from outside the device 300 into the device 300.
  • Figures 7A-7C show different views of sensor housing components 702 and sensor channels 306 of the device 300 of Figure 3A, in accordance with an exemplary embodiment.
  • Figure 7A is a first perspective view
  • Figure 7B is a second perspective view
  • Figure 7C is a side view.
  • the sensor housing components 702 couple together re each of the sensor channels 306 that may protrude through the apertures 305 on one side of the base housing 304.
  • the sensor housing components 702 include a plurality of apertures 703.
  • the sensor channels 306 may generally be hollow or have a hollow portion, and the hollow portions align with the respective apertures 703.
  • each of the sensor channels 306 include a slot or slit 704 at an end of the sensor channel 306 that protrudes through the apertures 305.
  • the slot 704 may be configured to hold the one or more binding moieties, which detect biomarkers in or on the user’s body.
  • the slots 704 house or contain the binding moieties, such as a protein, antibody, peptide, aptamer, or binding fragment thereof e.g., an scFvs, which generate a signal that is conveyed through the highly conductive graphene layers of the sensor channel 306 and/or the sensor housing components 702.
  • the hollow portion of the sensor channels 306 couple the slot 704 to the apertures
  • each sensor channel 306 is coupled to a circuit (for example, the circuit housed within the cavity 502, described below with reference to Figure 10) through the apertures 703, for example using nanowires or similar conductive or conducting structures.
  • the signals generated by the binding moieties in the slots 704 may be conveyed to the circuit.
  • the spatially resolvable electronic signal may manifest as a change in impedance or capacitance at the location of the bound biomarker.
  • each sensor housing component 702 couples thirty
  • sensor housing component 702 may couple together anywhere between 1 and 50 sensor channels 306. That is 1, 5, 10, 15, 20, 25, 30, 35, 40, or 50 sensor channels 306 may be coupled or an amount of sensor channels that is within a range defined by any two of the aforementioned number of sensor channels.
  • the sensor channels 306 are each formed from one or more graphene or graphene oxide layers, formed or printed as described herein or generated by similar processes.
  • the sensor channels 306 may be formed from some conductive or non-conductive material and have disposed on its surfaces the one or more graphene or graphene oxide layers.
  • the sensor housing components 702 may also comprise or be formed from one or more graphene or graphene oxide layers, formed or printed as described herein or generated by similar processes.
  • the binding moieties such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv
  • the binding moieties may be disposed within the slots 704 of the one or more sensor channels 306 as described herein.
  • the binding moieties such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv
  • the circuit is coupled directly to each of the sensor channels 306.
  • each sensor channel 306 coupled to the sensor housing components 702 may provide an individual signal to the circuit, thereby allowing for detection and/or monitoring of multiple biomarkers, etc., using a single device 300.
  • each of the sensor channels 306 may be isolated from each other sensor channel.
  • a first binding moiety such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv
  • a second binding moiety such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv
  • disposed with the slot 704b may generate a second signal in response to a second biomarker, thereby allowing the device 300 to specifically respond to or detect a plurality of different biomarkers within the single device 300.
  • Figure 8 shows a perspective view of the base housing 304 of Figure 5 combined with the sensor housing component 702 of Figures 7A-7C.
  • Figure 8 shows how the sensor channels 306 coupled to the sensor housing component 702 pass through the apertures 305 of the base housing 304.
  • the slots 704 are exposed at an exterior of the base housing 304 so that the slots 704 (including any binding moieties, etc.) are exposed to biofluids in the vicinity of the installed device 300.
  • the sensor housing components 702 move within the cavity 502, thereby allowing the sensor channels 306 to retract into or extend out of the base housing 304.
  • the sensor housing component 702 automatically retracts the sensor channels 306 when the device 300 is removed e.g., from the oral cavity or the tooth and may automatically extend the sensor channels 306 when the device 300 is e.g., attached to the oral cavity or the tooth.
  • the sensor housing component 702 and the sensor channels 306 are configured to move such that the sensor channels 306 extend from the device 302 such that the slots 704 are exposed to biofluids.
  • the sensor channels may retract back into the device 302.
  • the sensitive areas where the binding moieties, such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, are coupled to the slots 704 are protected from damage and/or contamination when not installed in, for example, the oral cavity, such as in the supra or sub-gingival space, preferably proximal to the cementoenamel-junction (CEJ), for instance coronal to the cementoenamel- junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth).
  • CEJ cementoenamel-junction
  • FIGS 9A and 9B show perspective views of a housing attachment 900 that attaches the device 300 of Figure 3A to a surface of a subject’s body, in accordance with an exemplary embodiment.
  • the housing attachment 900 comprises three surfaces as shown: a mounting base 902, a retaining member 904, and a cover 906.
  • the housing attachment 900 may generally allow the device 300 to be coupled to, for example, a tooth of the subject or in the supra or sub-gingival space, preferably proximal to the cementoenamel-junction (CEJ) (for instance coronal to the cementoenamel-junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth), in a removable fashion but ensure that the device 300, when installed on the tooth or in the supra or sub-gingival space, preferably proximal to the cementoenamel- junction (CEJ) (for instance coronal to the cementoenamel-j unction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth), it does not impact or impede mastication, speech, normal oral function etc., of the subject and ensures that the device 300 stays in place until the subject wants to remove the
  • the mounting base 902 couples the housing attachment 900 to the surface of the subject’s body, for example a surface of a tooth (similar to an individual tine of braces) or in the supra or sub-gingival space, preferably proximal to the cement-enamel-junction (CEJ).
  • the mounting base 902 is attached or otherwise coupled to the tooth via an adhesive or other similar compound or bonding agent.
  • the mounting base 902 includes a plurality of raised portions to which the device 300 may be coupled.
  • the raised portions of the mounting base 902 includes two levels, where a second level is taller or further raised than the first level, which presents as a step-like structure on the mounting base 902.
  • the second level may act as a physical stop or impediment to a structure or object moving along a surface of the first level.
  • a mounting structure of the device 300 moves along the surface of the first level until it butts against a side of the second level.
  • the retaining member 904 is a structure that allows the mounting structure of the device 300 to pass over the retaining member 904 but then generally keep the mounting structure of the device 300 in the passed over (or retained) location. Accordingly, the retaining member 904 is tapered to allow the mounting structure of the device 300 to slide into the retained location but prevent the mounting structure from easily moving out of the retained location. When the mounting structure of the device 300 is in the retained location, the device 300 may be held on the tooth with minimal movement until the user actively removes the device 300 from the housing attachment. This prevents the device 300 from moving and impeding mastication, speaking, etc., and keeps the device 300 safe from damage.
  • the cover 906 may cover the retaining member 904 and at least a portion of the raised portion(s) of the mounting base 902. Accordingly, as shown in Figures 9A and 9B, when assembled into the housing attachment 900, the mounting base 902, the retaining member 904, and the cover 906 create a channel within which the mounting structure of the device 300 rests when the device 300 is installed. In some embodiments, the cover 906 is coupled and held onto the raised portion of the mounting base 902 via one or more screws that pass-through holes 908 in the cover and couple to holes 910 in the mounting base 902.
  • FIG. 10 illustrates a block diagram of an aspect of a circuit 1000 coupled to the sensor channels 306 of the device 300 of Figure 3A, in accordance with an exemplary embodiment.
  • the circuit 1000 is housed within the device 300 such that it is in direct physical connection with the sensor channels 306 of the device.
  • the circuit 1000 is an example of a device that may be configured to implement the various methods described herein.
  • the circuit 1000 may receive or otherwise obtain signals generated by the binding moieties, such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, of each sensor channel 306 (for example, disposed within the slot 704 of each sensor channel 306).
  • the binding moieties such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv
  • the circuit 1000 may then analyze, store, or transmit the signal or information based on the signal. For example, when the device 300 is positioned within the subject’s mouth, the circuit 1000 may transmit the signals received from the binding moieties, such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, or information based on the signals to the subject’s cell phone, computer, tablet, or other electronic device for display or other consumption.
  • the binding moieties such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, or information based on the signals to the subject’s cell phone, computer, tablet, or other electronic device for display or other consumption.
  • the circuit 1000 may include a processor 1004, which controls operation of the circuit 1000.
  • the processor 1004 may also be referred to as a central processing unit (CPU).
  • Memory 1006 which may include both read-only memory (ROM) and random access memory (RAM), may provide instructions and data to the processor 1004.
  • a portion of the memory 1006 may also include non-volatile random access memory (NVRAM).
  • the processor 1004 typically performs logical and arithmetic operations based on program instructions stored within the memory 1006.
  • the instructions in the memory 1006 may be executable to implement the methods described herein.
  • the processor 1004 may comprise or be a component of a processing system implemented with one or more processors.
  • the one or more processors may be implemented with any combination of general-purpose microprocessors, microcontrollers, digital signal processors (DSPs), field programmable gate array (FPGAs), programmable logic devices (PLDs), controllers, state machines, gated logic, discrete hardware components, dedicated hardware finite state machines, or any other suitable entities that can perform calculations or other manipulations of information.
  • the processing system may also include machine-readable media for storing software.
  • Software shall be construed broadly to mean any type of instructions, whether referred to as software, firmware, middleware, microcode, hardware description language, or otherwise. Instructions may include code (e.g., in source code format, binary code format, executable code format, machine learning, or any other suitable format of code). The instructions, when executed by the one or more processors, cause the processing system to perform the various functions described herein. Accordingly, the processing system may include, e.g., hardware, firmware, and software, or any combination therein.
  • the instructions may cause the processing system to monitor the signals received from the binding moieties, such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, and store the signals in the memory 1006 or transmit the signals received.
  • the instructions may cause the processing system to process the signals received to identify and/or determine information from the signals received.
  • the instructions may cause the processing system to identify the host- microbial molecular information described above with reference to Figure 2.
  • the instructions may cause the processing system to monitor the signals received and determine whether the signals indicate any issues or concerns within the host’s body, for example whether the host is at risk to develop a disease, or the host is sick, has a particular disease or a condition associated with a disease or can allow monitoring of the state of the user during or after a therapy.
  • the instructions may cause the processing system to store monitored or determined information in the memory 1006.
  • the circuit 1000 may also include a housing 1008 that may include a transmitter
  • the circuit 1000 may include an antenna diversity system or array 1016 attached to the housing 1008 and electrically coupled to the transceiver 1014.
  • the antenna diversity system or array 1016 may include two antennas as an example, although more than two antennas or less than two antennas are also envisioned.
  • the circuit 1000 may also include multiple (wired or wireless) transmitters, receivers, transceivers, and/or antennas diversity systems.
  • the transmitter 1010 can be configured to transmit (via a wired or wireless connection) messages.
  • the processor 1004 may process messages and data to be transmitted via the transmitter 1010.
  • the receiver 1012 can be configured to receive messages (via the wired or wireless connection).
  • the processor 1004 may further process messages and data received via the receiver 1012.
  • the transmitter 1010 and the receiver 1012 may be configured to communicate via one or more communication protocols, including wired and wireless communications protocols (for example, LTE, 5G, Wi-Fi, Bluetooth, ZigBee, Z-Wave, ultrasound, or infrared, etc., protocols).
  • the circuit 1000 may also include a signal detector 1018 and/or a digital signal processor (DSP) that may be used in an effort to detect and quantify the level of signals or process the signals received by the transceiver 1014 or received from the sensor channels 306.
  • the signal detector and/or DSP 1018 may detect such signals as total energy, energy per subcarrier per symbol, power spectral density and other signals.
  • the signal detector and/or DSP 1018 may also be used to detect the signals received from the binding moieties, such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, in the sensor channels 306.
  • binding moieties such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, in the sensor channels 306.
  • details regarding the received signals, as provided by the signal detector and/or DSP 1018 are indicative of conditions within the body of the subject, etc.
  • the circuit 1000 may further comprise a user interface 1020 in some aspects.
  • the user interface 1020 may comprise one or more of a pushbutton interface, a microphone, a speaker, or light emitting diodes (FEDs, e.g., as a display), among others.
  • the user interface 1020 may include any element or component that conveys information to a user of the circuit 1000 and/or receives input from the user.
  • the circuit 1000 may also comprise one or more power sources 1022 and/or internal sensors 1024.
  • the power source 1022 may be configured to receive power wirelessly to charge an internal energy storage device or to power the circuit 1000.
  • the power source 1022 comprises the internal energy storage device and is configured to be charged when the device 300 is removed from the user’s mouth.
  • one or more devices 300 are coupled to the object in the shape of orthodontic bracket; when the orthodontic bracket is removed from the user’s mouth, the user may place or connect the orthodontic bracket on/to a charger that provides power to the power source 1022 or charges the power source 1022.
  • the bracket body has gingival base on the buccal and/or lingual surfaces.
  • the mesial, distal, occlusal surfaces are designed to prevent interference with adjacent anatomical hard and soft tissue.
  • the circuit 1000 of each of the one or more devices 300 is further configured to convey the received signals or information based on the signals to an external device (e.g., the user’s phone or other electronic device).
  • the power source 1022 may be shared among multiple circuits 1000 of multiple devices 300. For example, when multiple devices 300 are all coupled to a single object, such as the orthodontic bracket, the power source 1022 may be external to each device 300 but coupled to the circuit 1000 in each device. For example, the power source 1022 may be contained within the wire of the bracket or another retaining portion of the bracket (or object).
  • the circuit 1000 may also comprise one or more internal sensors 1024. In some aspects, the one or more internal sensors 1024 may be configured to provide information to the processor 1004 or any other component of the circuit 1000.
  • the various components of the circuit 1000 may be coupled together by a bus system 1026.
  • the bus system 1026 may include a data bus, for example, as well as, a power bus, a control signal bus, and a status signal bus in addition to the data bus.
  • a data bus for example, as well as, a power bus, a control signal bus, and a status signal bus in addition to the data bus.
  • Those of skill in the art will appreciate that the components of the circuit 1000 may be coupled together or accept or provide inputs to each other using some other mechanism.
  • processor 1004 may be used to implement not only the functionality described above with respect to the processor 1004, but also to implement the functionality described above with respect to the signal detector and/or DSP 1018. Further, each of the components illustrated in FIG. 10 may be implemented using a plurality of separate elements.
  • the various components of the device 300 may be formed from various materials and coated with one or more graphene or graphene oxide layers (for portions desired to be conductive) or an insulating material (for portions desired to be insulating or non- conductive).
  • the graphene or graphene oxide layers are disposed using a printing process, for example via a graphene or graphene oxide printer such as the SonoPlot MicroplotterTM.
  • the graphene or graphene oxide layers are disposed using a chemical process or an etching process or some similar method of disposing graphene or graphene oxide on a conductive or non-conductive surface.
  • the components of the device 300 are not formed from graphene or graphene oxide, they may be formed by any known method of manufacturing similar components and devices, such as via a 3D printer, molding, etc.
  • the base housing 304 is formed to have the apertures 305 and the sensor channels 306 are formed to include graphene or graphene oxide layers on at least the surfaces, the sensor channels 306 coupled to the sensor housing components 702.
  • the sensor channels 306 coupled to the sensor housing components 702 are placed within the cavity along with the circuit 1000.
  • the circuit 1000 is electrically and physically coupled to each of the sensor channels 306.
  • the circuit 1000 (and its corresponding components) is coupled to each of the binding moieties, such as a protein, antibody, peptide, aptamer, or binding fragment thereof e.g., an scFv, within the slots 704 of the sensor channels 306 such that any signals generated by the binding moieties, such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, are conveyed to the circuit 1000.
  • the cover 302 is then placed on the base housing 304 and pushed onto/into the base housing 304 such that the gasket 405 creates an impervious barrier between the cover 302 and the base housing 304 to keep everything out of the device 300.
  • the device 300 has one or more components constructed, at least in part, or coated based on graphene or graphene oxide technology that performs real time sensing and stores or communicates (for example, wirelessly) information received and/or determined based on one or more biofluids.
  • the information is stored and/or communicated according to an open platform or structure. For example, nanoprinting of graphene or graphene oxide on a surface of the object in the shape similar to orthodontic bracket, or tooth crown, or gingival implant is utilized as the basis of the sensing technology for the device on the tooth surface.
  • the device is configured for placement in the supra or sub-gingival space, preferably proximal to the cementoenamel- junction (CEJ) or coronal to the cementoenamel-junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth.
  • the graphene or graphene oxide printed structure may provide a low-cost, flexible, highly conductive, and water repellant structure that conveys “signals” between a sensing medium (e.g., the binding moieties) and a circuit.
  • the circuit 1000 may interpret received signals as detected biomarkers.
  • the circuit 1000 may be removable from the device 300 (for example, the orthodontic bracket) to transfer information regarding and/or based on the detection of the biomarkers.
  • the printing of graphene or graphene oxide monolayers is known in the art, see, for example, Mannoor et al., Graphene-Based Wireless Bacteria Detection on Tooth Enamel, Nature Communications, March 27, 2012, doi: 10.1038/ncomms 1767, which is hereby expressly incorporated by reference in its entirety and especially with respect to its disclosure of methods of printing graphene or graphene oxide layers.
  • binding moieties such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv
  • binding moieties such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv
  • binding moieties may comprise any binding moiety, such as a protein, peptide, antibody, aptamer, or binding fragment thereof e.g., an scFv, as is known in the art, which is capable of attaching to any biomarker.
  • binding moieties include but are not limited to receptors, ligand binding fragments, antibodies and binding fragments thereof including polyclonal antibodies or monoclonal antibodies or binding fragments thereof, single chain variable regions (scFv’s), T- cell receptors and fragments thereof, peptides, antimicrobial or surface-active peptides, lectins, polysaccharides, nucleic acids, aptamers, modified nucleic acids, peptide-nucleic acids, 5’-0- methyl ribonucleic acids (RNAs), 5’-fluoro-RNAs, or 5’-0-ethyl RNAs and DNA oligonucleotides.
  • scFv single chain variable regions
  • Derivatization of said derivatizable graphene or graphene oxide surface may be carried out by many approaches, such as, for example, chemical modification of one or both surfaces of said graphene as with hydroxyl, epoxy, thiol, amino, or ester groups, or other chemical moieties known to be useful in the attachment of peptides, proteins, nucleic acids, or small molecule ligands to surfaces.
  • Said derivatization may further be carried out by utilizing graphene or graphene oxide binding peptides as described in Mannoor et al., Nature Communications, volume 3, Article number: 763 (2012), which is hereby expressly incorporated by reference in its entirety, especially with respect to its disclosure of the generation of graphene-based electrosensors and of the use of graphene binding peptides therein.
  • Said derivatizable graphene or graphene oxide surfaces may be attached to a substrate, wherein said substrate may comprise a ceramic layer, a plastic layer, a glass layer, a metallic layer, a metal-oxide layer, a titanium layer, a composite resin layer, and/or one or more polymer layers.
  • a coil antenna coupled to the graphene or graphene oxide layer may transmit signals and/or information representative of the binding to the specific antigens through wireless transmission; alternatively, the signals and/or information representative of the binding to the specific antigens may be transmitted via wired communication.
  • Proteins, antibodies, peptides, aptamers, or binding fragments thereof e.g., scFvs can be incorporated, integrated, associated, or embedded into graphene or graphene oxide using supramolecular chemistry or supramolecular bonding and other methods known in the art.
  • Binding of a protein, antibody, peptide, aptamer, or binding fragment thereof e.g., an scFv to graphene or graphene oxide can be covalent or non-covalent.
  • Proteins, antibodies, peptides, aptamers, or binding fragments thereof e.g., an scFv can be coupled to the graphene or graphene oxide through disulfide, maleimide chemistry, NHS chemistry using N-hydroxysuccinimide esters, for example, or other amine-reactive groups, for example.
  • Exemplary chemical groups that bond with amines include, but are not limited to, isothiocyanates, isocyanates, acyl azides, NHS, sulfonyl chlorides, aldehydes, glyoxals, epoxides, oxiranes, carbonates, aryl halides, imidoesters, carbodiimides, anhydrides, or fluorophenyl esters.
  • Conjugation to amines can be by acylation or alkylation.
  • Functional groups that condense amines include carbonyl groups, for example.
  • Functionalization of graphene or graphene oxide can be performed using carboxylates, aminopropyltriethoxysilane (APTS), or 3-methylimidazolium bromide, for example.
  • Pyrene-NHS ester (Pyr-NHS) molecules can be used as linkers to incorporate, integrate, or associate proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFvs to the graphene or graphene oxide support provided in the devices and systems set forth in this disclosure.
  • the linker can retain a succinimidyl ester group, which interacts with the amino groups provided on the proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFvs to generate a stable, amide bond.
  • the Pyr-NHS can have an aromatic pyrenyl group, which can strongly interact with the graphene surface, via non- covalent p-p stacking. Noncovalent interaction can also include hydrophobic, hydrophilic, electrostatic and van der Waals interactions.
  • the Pyr-NHS ester can cause the pyrene group at one end of the linker to strongly bind to the graphene surface via p-p interactions and the succinimidyl ester group can covalently react with the amino group (NH2) of the proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFvs.
  • Pyr-NHS molecules can be applied to cover the complete surface of the graphene, a region of the surface of the graphene, or a portion of the surface of the graphene.
  • Proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains can be incubated at low temperature with the graphene under conditions supporting binding for at least 15 minutes, 30 minutes, 1 hour, two hours, three hours, four hours, five ours, six hours, seven hours, eight hours, nine hours, ten hours, eleven hours, twelve hours, 18 hours, 24 hours or longer or for a time that is within a range defined by any two of the aforementioned time points, for example, followed by rinsing with a buffer such as PBS, HEPES, or Tris and dried to generate the graphene support having the proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains.
  • Graphene oxide can be used in some embodiments as the support in the devices and systems described herein for the proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains. Proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains can be attached to the graphene oxide using approaches similar to that described above.
  • graphene oxide surfaces can be incorporated, integrated, or associated with the proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains described herein using“click” chemistry.“Click” chemistry can be used in combination with a sortase-mediated transpeptidation reaction.
  • “click” chemistry.“Click” chemistry can be used in combination with a sortase-mediated transpeptidation reaction.
  • graphene oxide can be functionalized with carboxylic acids to facilitate incorporation, integration, or association of the proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains.
  • the carboxylic group on graphene oxide can be activated with N- hydroxysuccinimide (NHS) in a reaction catalyzed by l-Ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride (EDC).
  • EDC l-Ethyl-3-(3- dimethylaminopropyl)carbodiimide hydrochloride
  • a diamino-polyethylane glycol polyethylene glycol (PEG) linker can be introduced, with one end reacting with the N- hydroxysuccinimide (NHS) moiety.
  • the other end of the PEG linker is further functionalized with an NHS-activated dibenzocycolctyne (DBCO), for example.
  • DBCO NHS-activated dibenzocycolctyne
  • Graphene or graphene oxide sheets can be prepared by any method, including bioprinting. Any printer suitable for bioprinting or three-dimensional (3D) printing can be used. As an example, a sonoplot Microplottor Proto printer can be used.
  • the graphene or graphene oxide sheets can range in thickness from 10mM to 0.55mm.
  • the graphene or graphene oxide sheet can have a thickness that is at least 10mM, 20mM, 30mM, 40mM, 50mM, 60mM, 70 mM, 80mM, 90mM, 100 mM, I IOmM, 120mM, 130mM, 140mM, 150mM, 160mM, 170mM, 180mM, 190 mM, 200mM, 210mM, 220mM, 230mM, 240mM, 250mM, 260mM, 270mM, 280mM, 290mM, 300mM, 310mM, 320mM, 330mM, 340mM, 350mM, 360mM,370mM,380mM,390mM, 400mM, 410mM, 420mM, 430mM, 440mM, 450mM, 460mM, 470 mM, 480mM, 490mM, 500mM, 510mM,
  • the present disclosure contemplates a mesh-like graphene structure with an ability to evaluate multi-parametric properties, including the concentrations and amounts of biomarkers in bodily fluids.
  • multi-parametric properties including the concentrations and amounts of biomarkers in bodily fluids.
  • Such evaluation of multi- parametric properties may result from both detecting individual binding and detecting the unique signatures derived from multiplex binding of multiple biomarkers.
  • Raman spectroscopic evaluation of functionalized graphene may be used to determine changes in graphene surfaces with continuous exposure of multiplex antigens.
  • the methods, devices and systems of the present disclosure may be applied in conjunction with an in vitro system with capabilities to sense the molecular changes of one marker perturbation within a wet microenvironment.
  • a derivatizable, electrically conductive surface is provided, wherein binding moieties, such as proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains, for biomarkers indicative of oral and/or general health are provided in a spatially resolved manner.
  • said derivatizable, electrically conductive surface may comprise a metal surface, a semiconductive surface, ceramic, a polymer, or a graphene or graphene oxide layer.
  • the device 300 described herein may attach to a tooth or tooth structure of a host and transmit or store signals while housing (and protecting) an internal component (such as the sensor channels 306 and circuit 1000).
  • the device 300 is similar to an orthodontic bracket component that is easy to access and use by the host while providing continuous detection and transmission of information based on oral biofluids while avoiding disturbances to and from mastication and speaking.
  • multiple devices 300 can be inserted into the host’s mouth for detection of multiple of the same parameters.
  • the device is configured for placement in the supra or sub-gingival space, preferably proximal to the cementoenamel-j unction (CEJ) or coronal to the cementoenamel-junction following the normal contour of the gingival tissue at the CEJ, and respecting but not invading the biological width between the gingiva, alveolar bone and tooth.
  • CEJ cementoenamel-j unction
  • the integrative sensing tools described herein capture the complexity of the human saliva.
  • the device 300 and methods described herein enable individual and population detection in real-time. With a combination of the graphene based sensing components of the device 300, and functionalization of detection moieties, the device 300 is able to communication information regarding the human saliva to external devices, for example via wireless signals. In real-time, the device 300 will simultaneously and selectively measure metabolites (e.g., glucose and lactate) and electrolytes (e.g., sodium and potassium ions), as well as skin temperature for signal calibration. Such model bridges existing technological gaps among signal transduction, conditioning, processing and wireless transmission in in vivo biosensors.
  • metabolites e.g., glucose and lactate
  • electrolytes e.g., sodium and potassium ions
  • the device 300 and corresponding methods described herein have the potential to change science (data collection) and patient care (diagnosis, response to treatment) relative to in vivo sensing.
  • the device 300 may measure five different markers from the host’s immune response; from the microbiome where the device 300 is placed, and from the host’s or device 300 position’s metabolism.
  • the sensing structures of the device 300 may be highly sensitive, stable and repeatable for saliva (or other biofluid) analysis.
  • the device 300 may operate or be used in singleplex or in multiplex modes to monitor dynamic changes in real-time.
  • the device 300 is able to overcome major challenges in in vivo biosensors for monitoring of salivary biomarkers due to its ability to be sensitive and selective and obtain accurate salivary profiles.
  • Given complexities of saliva secretion, simultaneous, multiplexed screening of target biomarkers by the device 300 is a major benefit over the art and allows for seamless full system integration.
  • Such sensitivity and selectivity as described herein provide benefits for at least the following:
  • Glucose and lactate levels Their detection can be achieved using the device 300 utilizing highly sensitive and selective graphene-based electrochemical enzymatic biosensors. Considering that the glucose levels in human saliva are very low (tens of mM), commercially available gold and carbon electrodes cannot be used for direct saliva analysis due to the lack of sensitivity.
  • the device 300 may provide the highly sensitive and selective and glucose and lactate sensors through the combination of the high-performance carbon nanomaterials (e.g., laser scribed graphene and enzymatic reactions).
  • LPS Lipopolisacharide
  • the analysis of LPS and insulin is performed using the enzyme linked immunosorbent assays (ELISA) in both research and clinical settings. While these assays offer benefits including high sensitivity, selectivity and a wide concentration detection range, these assays are inherently discrete in terms of both implementation as well as obtained data.
  • the device 300 may combine graphene or graphene oxide electrodes with molecularly imprinted polymers (MIPs) for dynamic biosensing. MIPs may be tailor- made recognition materials that can mimic biological receptors.
  • MIPs molecularly imprinted polymers
  • recognition units for biosensor fabrication they may outperform natural receptors in their durability, chemical stability, and low production costs. More importantly, they can be easily recovered and are suitable for dynamic measurements.
  • An epitope approach - a representative fragment of target molecule - is applied to prepare templates for MIPs.
  • a binding capacity of surface imprinted artificial receptors may be maximized by increasing the effective surface area through the additional graphene films before the formation of the MIPs.
  • Functional monomers and cross-linkers that also be optimized to achieve higher sensitivity.
  • a redox probe (such as Prussian blue) may be used between the MIP and graphene or graphene oxide structure.
  • the bonding of target molecules with MIP and blocking of the pathway of solution to the redox probe then result in a decreased redox signal from the biosensor of the device 300.
  • the sensing platform utilizing the device 300 can be an excellent candidate for ultra-low-power and miniaturized biosensors.
  • CRP C-reactive protein
  • Acute phase c-reactive protein is regulated by most known inflammatory cytokines including interleukin- 1,-6 and tumor necrosis alpha.
  • the device 300 may operate to detect levels of CRP.
  • MIP based approach could be ideal for detecting small molecules such as LPS and insulin, due to the thick morphology of bulk imprints, big template molecules such as CRP could be difficult to detect via conventional methods as they are embedded in the matrices too deeply that lead to restriction or no access for the target molecules to bind to sensors, which may be resolved by the biosensors of the device 300. In this case, in order to achieve label- free and highly sensitive detection of the key inflammatory biomarker CRP.
  • the continuous data obtained via the in vivo devices 300 may be analyzed using machine learning, artificial intelligence, statistical, analytical, or visualization methods or models.
  • the device 300 may various problems, including how to monitor multiplexed biomarkers in whole saliva simultaneously via a single integrated device and how to obtain/implement a reliable integrated system that could be potentially used for further in vivo applications by providing for an in vivo sensor array that can accurately and consistently measure the concentrations of biomarkers in saliva that is able to determine whether the concentrations measured in a subject’s saliva provide accuracy when in combinatorial analysis in real-time.
  • the device 300 may be used alone to monitor individual molecular changes of biomarkers, for example, in saliva. Such individual monitoring may provide information regarding crosstalk between immune-microbiome-metabolism in the context of various diseases and/or conditions.
  • the device 300 accordingly, may be configured to sense molecular changes of one marker perturbation in various microenvironments (for example, the mouth).
  • the device 300 may be used in combination with other devices 300 and/or components to create a system to monitor multiplex molecular changes of biomarkers. By allowing the devices 300 to transmit information while functioning, the devices 300 may perform multiparametric sensing in vitro.
  • the device 300 may provide for detection potential to improve individual and populational oral monitoring and may provide for the capture of host-microbial metabolites that are modulated by human microbiome and metabolism, with important health implications for the host immune response.
  • the device 300 and corresponding methods may provide benefits in the maintenance of long-term health, pre-disease disease transitions, and the development of new therapeutics.
  • the device 300 may also provide for detection of oral markers related to a systemic condition with high incidence; monitoring of functional changes of saliva to understand unknown host-microbial-metabolic patterns at the individual level; investigating in real-time predictors of disease state; moving beyond assessing only sugar levels of chronic inflammatory conditions; and addressing multiplex capacity of saliva detection.
  • the biosensor used in the device 300 may be capable of functioning with and detecting changes and biomarkers in small amounts of biofluids, for example less than 10 pL.
  • the biomarkers which are detectable by this system may comprise any biomarker which is present in a biofluid for which a binding moiety such as, a protein, antibody, aptamer, or binding fragment thereof e.g., scFv domain, is known or could readily be developed by one of skill in the art.
  • Said biomarkers may include but are not limited to immune or inflammatory biomarkers such as inflammatory c-reactive protein (CRP), microbiome-derived biomarkers such as lipopolysaccharides (LPS), glucose, lactate Erv-l , and metabolically derived biomarkers or hormones, such as cortisol hormone.
  • CRP inflammatory c-reactive protein
  • LPS lipopolysaccharides
  • glucose lactate Erv-l
  • metabolically derived biomarkers or hormones such as cortisol hormone.
  • Methods of using the devices described herein are useful for biomarker characterization e.g., to detect or monitor temporal release and uptake rates of molecules or physical, chemical, or functional changes in biomarker presence or concentration in bodily fluids.
  • Fluorescently labelled compounds may also be used to indicate or calibrate electrical resistance changes on the devices or systems described herein so as evaluate the binding and unbinding of molecules from the graphene or graphene oxide surface on the device or system.
  • the devices or systems described herein are configured, e.g., by virtue of incorporation, integration or association of the proteins, peptides, antibodies, aptamers, or binding fragments thereof e.g., scFv domains into said graphene or graphene oxide components of said devices or systems, so as to detect biomarkers for inflammation, diseases associated with inflammation, or disease states or conditions associated with inflammation including, but not limited to, allergic reactions, anaphylactic reactions, arthritis, asthma, atherosclerosis, bone diseases, breast cancer, cancer, cardiovascular diseases, colon cancer, degenerative neurologic disorders, dementia, diabetes mellitus, eye diseases, gastrointestinal disorders, genitourinary disorders, hematologic disorders, hepatobiliary disorders, hypertension, infectious diseases, leukemia/lymphoma, lung cancer, metabolic disorders, neurological disorders, neuromuscular disorders, obesity /eating disorders, parasitic diseases, perinatal disorders, pregnancy, prostate cancer, psychiatric disorders, pulmonary disorders, renal disorders,
  • Some embodiments of the devices or systems described herein incorporate, integrate, or associate into the graphene or graphene oxide component of said devices or systems at least one ERV-l detecting antibody, such as that set forth in a molecule comprising a sequence that comprises at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, and/or 16, or a biologically active variant or binding fragment thereof, or any combination thereof.
  • the systems or devices described herein comprise at least one ERV-l detecting antibody (e.g., resolvimab) or a binding fragment thereof e.g., an scFv incorporated, integrated or associated into the graphene or graphene oxide component of said devices or systems.
  • the systems or devices described herein comprise a ERV-l detecting antibody or fragment thereof, which is a biomimetic of the resolvin class of lipid molecules.
  • the resolvimab antibodies or binding fragments thereof specifically recognize, bind to, or associate with ERV-l.
  • the term “ERVl” is used interchangeably with ChemR23, ChemerinR, and RVER1.
  • EVR1 is a G protein-coupled receptor Chemokine like receptor 1 expressed in innate immune cells (neutrophils and monocytes) and most cells of the human body.
  • the lipid mediator resolvin El (RvEl) is known to activate cell signaling through the ERVl receptor (Arita et al. (2005) J. Exp Med 201 (5):7l 3-722).
  • the resolvimab incorporated, integrated, or associated with the graphene or graphene oxide component of said systems or devices described herein comprises a variable heavy chain (VH) sequence comprising an amino acid sequence that comprises at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 6, 10, and 14, or biologically active variant or binding fragment thereof, or any combination thereof.
  • VH variable heavy chain
  • the resolvimab incorporated, integrated, or associated with the graphene or graphene oxide of said system or device described herein comprises a variable light chain (VK) sequence comprising an amino acid sequence that comprises at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 4, 8, 12, and 16, or a biologically active variant or binding fragment thereof, or any combination thereof.
  • VK variable light chain
  • the resolvimab (e.g ., 1G12.G8-2.C3) incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises a variable heavy chain (VH) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 2 and a variable light chain (VK) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 4.
  • VH variable heavy chain
  • VK variable light chain
  • the resolvimab (e.g., 1G12.G8- 2.C3) incorporated, integrated, or associated with the graphene or graphene oxide component of a system or device described herein comprises a variable heavy chain (VH) sequence obtained from a nucleotide sequence comprising a nucleic sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequence set forth in SEQ ID NO: 1, which encodes a polypeptide sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 2, and a variable light chain (VK) sequence obtained from a nucleotide sequence comprising a nucleic sequence that is at least 70%, 75%
  • the resolvimab (e.g ., 5G4.F8-1.G5) incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises a variable heavy chain (VH) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 6 and a variable light chain (VK) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 8.
  • VH variable heavy chain
  • VK variable light chain
  • the resolvimab (e.g., 5G4.F8-1.G5) incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises a variable heavy chain (VH) sequence obtained from a nucleotide sequence comprising a nucleic sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequence set forth in SEQ ID NO: 5, which encodes a polypeptide sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 6, and a variable light chain (VK) sequence obtained from a nucleotide sequence comprising a nucleic sequence that is at least 70%, 75%,
  • the resolvimab (e.g., 5G4.F8-1B7) incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises a variable heavy chain (VH) sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 10 and a variable light chain (VK) sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence to the amino acid sequence set forth in SEQ ID NO: 12.
  • VH variable heavy chain
  • VK variable light chain
  • the resolvimab (e.g., 5G4.F8-1B7) incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises a variable heavy chain (VH) sequence obtained from a nucleotide sequence comprising a nucleic sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequence set forth in SEQ ID NO: 9, which encodes a polypeptide sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 10, and a variable light chain (VK) sequence obtained from a nucleotide sequence comprising a nucleic acid sequence that is at least 70%, 75%, 80%, 85%, 90%,
  • the resolvimab (e.g ., 5G4.F8-1.C10) incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises a variable heavy chain (VH) sequence comprising an amino acid that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 14 and a variable light chain (VK) sequence comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 16.
  • VH variable heavy chain
  • VK variable light chain
  • the resolvimab (e.g., 5G4.F8-1.C10) incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises a variable heavy chain (VH) sequence obtained from a nucleotide sequence comprising a nucleic sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the nucleic acid sequence set forth in SEQ ID NO: 13, which encodes a polypeptide that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 14, and a variable light chain (VK) sequence obtained from a nucleotide sequence comprising a nucleic acid sequence that is at least 70%, 75%, 80%, 85%, 90%,
  • ERV-l detecting antibody e.g., resolvimab
  • binding fragments thereof which can be incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein are described in WO 2018/213592, which is hereby expressly incorporated by reference herein in its entirety.
  • Table 1 provides the sequences for SEQ ID Nos. 1-16.
  • the antibody or binding fragment thereof e.g., scFv incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises an antibody or binding fragment thereof e.g., scFv specific for or that binds to or associates with glucose, lactate, c-reactive protein (CRP), or microbiome-derived biomarkers such as lipopolysaccharides (LPS), or insulin.
  • an antibody or binding fragment thereof e.g., scFv specific for or that binds to or associates with glucose, lactate, c-reactive protein (CRP), or microbiome-derived biomarkers such as lipopolysaccharides (LPS), or insulin.
  • CRP c-reactive protein
  • LPS lipopolysaccharides
  • the antibody or binding fragment thereof e.g., scFv incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises an antibody or binding fragment thereof e.g., scFv specific for or that binds to or associates with glucose and such an antibody for the detection of glucose is commercially available and can be obtained from a Glucose Assay Kit (Abeam, clone ab65333), as described in Jiang S et al. Let-7 Suppresses B Cell Activation through Restricting the Availability of Necessary Nutrients; Cell Metab 27:393-403. e4 (2018).
  • Additional antibodies for detecting glucose which can be incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein is a Mouse Glucose ELISA Kit (MyBioSource.com, MBS7200879). Binding fragments and scFv fragments of said antibodies can be generated by conventional approaches.
  • the antibody or binding fragment thereof e.g., scFv incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises an antibody or binding fragment thereof e.g., scFv specific for or that binds to or associates with lactate are commercially available and can be obtained from a L-Lactate Assay Kit (Colorimetric) (ab6533l) from Abeam, as described in Jeong JH et al. Intracellular glycolysis in brown adipose tissue is essential for optogenetically induced nonshivering thermogenesis in mice. Sci Rep 8:6672 (2018) and Menk AV etal. Early TCR Signaling Induces Rapid Aerobic Glycolysis Enabling Distinct Acute T Cell Effector Functions; Cell Rep 22: 1509-1521 (2016). Binding fragments and scFv fragments of said antibodies can be generated by conventional approaches.
  • the antibody or binding fragment thereof e.g., scFv incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises an antibody or binding fragment thereof e.g., scFv specific for or that binds to or associates with CRP, e.g., clone 5A9, which can be obtained from Novus Biologicals, as described in Kreutz RP et al., C-reactive protein and fibrin clot strength measured by thrombelastography after coronary stenting; Blood Coagul Fibrinolysis 2013 Apr; 24(3): 321-326. Binding fragments and scFv fragments of said antibodies can be generated by conventional approaches.
  • the antibody or binding fragment thereof e.g., scFv incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises an antibody or binding fragment thereof e.g., scFv specific for or that binds to or associates with LPS, e.g., clone 2D7/1, can be obtained from Novus Biologicals, as described in McCallus DE and Norcross NL, Antibody specific for Escherichia coli J5 cross-reacts to various degrees with an Escherichia coli clinical isolate grown for different lengths of time; Infect Immun. 1987 May; 55(5): 1042-1046.
  • Binding fragments and scFv fragments of said antibodies can be generated by conventional approaches.
  • the antibody or binding fragment thereof e.g., scFv incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein comprises an antibody or binding fragment thereof e.g., scFv specific for or that binds to or associates with insulin, e.g., clone ab2000l 1, can be obtained from Abeam, as described in Lijie Zhao, Tingli Sun, Lina Wan, Chitosan oligosaccharide improves the therapeutic efficacy of sitagliptin for the therapy of Chinese elderly patients with type 2 diabetes mellitus; Therapeutics and Clinical Risk Management June 27, 2017. Binding fragments and scFv fragments of said antibodies can be generated by conventional approaches.
  • the systems and devices described herein can be used in a variety of methods to detect one or more biomarkers available in a biofluid, preferably an oral biofluid of a subject and, in some embodiments, provide for a non-surgical, non-invasive and a safe approach to detect, diagnose, or monitor a disease in a subject, such as a human or an animal (e.g., cattle, pigs, sheep, horses, dogs, or cats), preferably inflammation, such as chronic inflammation, or a disease state or condition associated with inflammation or cancer.
  • a subject such as a human or an animal (e.g., cattle, pigs, sheep, horses, dogs, or cats), preferably inflammation, such as chronic inflammation, or a disease state or condition associated with inflammation or cancer.
  • the methods are practiced by contacting a subject, such as a human or animal (e.g., cattle, pigs, sheep, horses, dogs, or cats) with a system or device, as described herein, e.g., by contacting the subject (e.g., the flesh or oral mucosa) with said system or device.
  • a system or device as described herein is contacted or placed in the gingival crevice or periodontal pocket of the subjects’ oral cavity in a position where the system or device can come into contact with an oral biofluid such as saliva, crevicular fluid, gingival fluid, or oral sweat.
  • the device is placed in contact with the subject by placing the device at or near the supra or sub-gingival space, preferably proximal to the cement-enamel-junction (CEJ).
  • Biomarkers present in the oral biofluid such as Erv-l, glucose, lactate, CRP, LPS, or insulin are then detected by the antibody or binding fragment thereof e.g., scFv incorporated, integrated, or associated with the graphene or graphene oxide component of the system or device described herein and a signal is generated and transmitted to a receiver, preferably wirelessly.
  • biomarkers present a biofluid such as saliva, crevicular fluid, gingival fluid, or sweat of a subject
  • these methods of detection and diagnosis can be performed before or during traditional therapy of a subject having an inflammatory disease or disorder or cancer.
  • the methods of using the device described herein can also be continued periodically in a subject showing signs of recurrence of a disease or disorder.
  • the methods can be performed during a therapy for a disease or disorder or the methods can be practiced in a prophylactic context.
  • diagnosis using the device will be performed at a period of time effective in bringing about a desired result or outcome, be it prophylactic or initiation or continuation of therapeutic treatment of an inflammatory disorder or a neoplastic disease and/or symptoms associated therewith.
  • the methods of using the device can be practiced subsequent to, preceding, or contemporaneously with administration of therapies including therapies that elicit an immune response in the subject.
  • the subject may have previously been given or presently received chemotherapy, radiation therapy, CAR T cell therapy, or immunotherapy.
  • diagnosis is performed after an initial therapy or treatment, and the subject monitored for an immunological and/or clinical response.
  • diagnosis is performed after one or more administration of therapy or treatment subsequent to the initial therapy or treatment as appropriate, typically on a monthly, semimonthly, or preferably a weekly basis, until the desired result is observed. Thereafter, additional diagnoses can be performed as required, particularly when the immunological or clinical benefit from therapy or treatment appears to subside.
  • the system or device described herein is used to identify or monitor health, or a disease or condition associated with a disease detectable by identifying the presence or concentration of a biomarker indicative of the presence or severity of the disease or condition associated with the disease in a biofluid such as saliva, crevicular fluid, gingival fluid, or oral sweat, preferably an oral biofluid.
  • a biofluid such as saliva, crevicular fluid, gingival fluid, or oral sweat, preferably an oral biofluid.
  • Such diseases or conditions associated with a disease detectable using a system or device described herein include allergic reactions, anaphylactic reactions, arthritis, asthma, atherosclerosis, bone diseases, breast cancer, cancer, cardiovascular diseases, colon cancer, degenerative neurologic disorders, dementia, diabetes mellitus, eye diseases, gastrointestinal disorders, genitourinary disorders, hematologic disorders, hepatobiliary disorders, hypertension, infectious diseases, leukemia/lymphoma, lung cancer, metabolic disorders, neurological disorders, neuromuscular disorders, obesity /eating disorders, parasitic diseases, perinatal disorders, pregnancy, prostate cancer, psychiatric disorders, pulmonary disorders, renal disorders, rheumatic diseases, stroke, wound healing, oral infections, periodontal disease, brain injury, trauma or neuronal inflammation.
  • the systems and devices described herein are used in methods to detect or monitor a cancer or the progress of a therapy for cancer, wherein said cancer can be acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, anal cancer, appendix cancer, astrocytomas, atypical teratoid/rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer (osteosarcoma and malignant fibrous histiocytoma), brain stem glioma, brain tumors, brain and spinal cord tumors, breast cancer, bronchial tumors, Burkitt lymphoma, cervical cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-Cell lymphoma, embryonal tumors, endometrial cancer, ependymoblastoma, ependymo
  • the systems and devices described herein are used in methods to detect glucose, lactate, C-reactive protein (CRP), LPS, insulin, or any combinations thereof. In some embodiments, the systems and devices described herein are used in methods to detect ERV-l .
  • the molecular content of saliva is derived from combination of fluids present in the oral tissues: GCF, serum, interstitial fluids, water and oral sweat.
  • GCF fluids present in the oral tissues
  • serum fluids present in the oral tissues
  • interstitial fluids water and oral sweat.
  • continuous salivary monitoring has not been developed yet.
  • Nanowires and graphene technology presents mechanical, electrical and sensing properties in wet environments.
  • a graphene based oral device is first designed to continuously monitor biological changes. See Figure 1.
  • an interface may refer to hardware or software configured to connect two or more devices together.
  • an interface may be a part of a processor or a bus and may be configured to allow communication of information or data between the devices.
  • the interface may be integrated into a chip or other device.
  • an interface may comprise a receiver configured to receive information or communications from a device at another device.
  • the interface e.g., of a processor or a bus
  • an interface may comprise a transmitter configured to transmit or communicate information or data to another device.
  • the interface may transmit information or data or may prepare information or data for outputting for transmission (e.g., via a bus).
  • determining may include calculating, computing, processing, deriving, investigating, looking up (e.g., looking up in a table, a database or another data structure), ascertaining and the like. Also, “determining” may include receiving (e.g., receiving information), accessing (e.g., accessing data in a memory) and the like. Also,“determining” may include resolving, selecting, choosing, establishing and the like. Further, a“channel width” as used herein may encompass or may also be referred to as a bandwidth in certain aspects.
  • a phrase referring to“at least one of’ a list of items refers to any combination of those items, including single members.
  • “at least one of: a, b, or c” is intended to cover: a, b, c, a-b, a-c, b-c, aa, bb, cc, and a-b-c.
  • DSP digital signal processor
  • ASIC application specific integrated circuit
  • FPGA field programmable gate array
  • PLD programmable logic device
  • IC integrated circuit
  • a processing system may be implemented using one or more ICs or may be implemented within an IC (e.g., as part of a system on a chip).
  • the IC may comprise a general purpose processor, a DSP, an ASIC, an FPGA or other PLD, discrete gate or transistor logic, discrete hardware components, electrical components, optical components, mechanical components, or any combination thereof designed to perform the functions described herein, and may execute codes or instructions that reside within the IC, outside of the IC, or both.
  • a general purpose processor may be a microprocessor, but in the alternative, the processor may be any commercially available processor, controller, microcontroller or state machine.
  • a processor may also be implemented as a combination of computing devices, e.g., a combination of a DSP and a microprocessor, a plurality of microprocessors, one or more microprocessors in conjunction with a DSP core, or any other such configuration.
  • the functions described may be implemented in hardware, software, firmware, or any combination thereof. If implemented in software, the functions may be stored on or transmitted over as one or more instructions or code on a computer-readable medium.
  • Computer-readable media includes both computer storage media and communication media including any medium that facilitates transfer of a computer program from one place to another.
  • a storage media may be any available media that can be accessed by a computer.
  • such computer-readable media can comprise RAM, ROM, EEPROM, CD-ROM or other optical disk storage, magnetic disk storage or other magnetic storage devices, or any other medium that can be used to carry or store desired program code in the form of instructions or data structures and that can be accessed by a computer.
  • any connection is properly termed a computer-readable medium.
  • the software is transmitted from a website, server, or other remote source using a coaxial cable, fiber optic cable, twisted pair, digital subscriber line (DSL), or wireless technologies such as infrared, radio, and microwave
  • the coaxial cable, fiber optic cable, twisted pair, DSL, or wireless technologies such as infrared, radio, and microwave are included in the definition of medium.
  • Disk and disc includes compact disc (CD), laser disc, optical disc, digital versatile disc (DVD), floppy disk, and Blu-ray® disc where disks usually reproduce data magnetically, while discs reproduce data optically with lasers.
  • computer readable medium may comprise non-transitory computer readable medium (e.g., tangible media).
  • computer readable medium may comprise transitory computer readable medium (e.g., a signal). Combinations of the above should also be included within the scope of computer-readable media.
  • certain aspects may comprise a computer software product for performing the operations presented herein.
  • a computer program product may comprise a computer readable medium having instructions stored (and/or encoded) thereon, the instructions being executable by one or more processors to perform the operations described herein.
  • the computer software product may include packaging material.
  • the methods disclosed herein comprise one or more steps or actions for achieving the described method.
  • the method steps and/or actions may be interchanged with one another without departing from the scope of the claims.
  • the order and/or use of specific steps and/or actions may be modified without departing from the scope of the claims.
  • modules and/or other appropriate means for performing the methods and techniques described herein can be downloaded and/or otherwise obtained by a user terminal and/or base station as applicable.
  • a user terminal and/or base station can be coupled to a server to facilitate the transfer of means for performing the methods described herein.
  • various methods described herein can be provided via storage means (e.g., RAM, ROM, a physical storage medium such as a compact disc (CD) or floppy disk, etc.), such that a user terminal and/or base station can obtain the various methods upon coupling or providing the storage means to the device.
  • storage means e.g., RAM, ROM, a physical storage medium such as a compact disc (CD) or floppy disk, etc.
  • CD compact disc
  • floppy disk etc.
  • any other suitable technique for providing the methods and techniques described herein to a device can be utilized.

Abstract

La présente invention concerne une pluralité de dispositifs ayant des biocapteurs à base d'oxyde de graphène et de graphène, qui sont configurés pour détecter, surveiller et suivre des informations de santé d'un sujet, tel qu'un être humain ou un animal, telles que déterminées par la détection de biomarqueurs hôtes-microbiens-métaboliques disponibles dans des fluides biologiques, tels que la salive et le fluide créviculaire gingival.
PCT/US2019/020559 2018-03-05 2019-03-04 Système et procédé de détection sans fil d'état de santé buccale WO2019173220A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201862638793P 2018-03-05 2018-03-05
US62/638,793 2018-03-05

Publications (1)

Publication Number Publication Date
WO2019173220A1 true WO2019173220A1 (fr) 2019-09-12

Family

ID=67847432

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2019/020559 WO2019173220A1 (fr) 2018-03-05 2019-03-04 Système et procédé de détection sans fil d'état de santé buccale

Country Status (1)

Country Link
WO (1) WO2019173220A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20210293776A1 (en) * 2020-03-18 2021-09-23 Nanodx, Inc. Measurement systems and associated techniques for sensing electrical characteristics of a sensor
WO2022232373A1 (fr) * 2021-04-28 2022-11-03 Freire Marcelo Système et procédé de biodétection immunitaire et moléculaire par l'intermédiaire d'une brosse à dents
RU220817U1 (ru) * 2023-07-10 2023-10-04 Федеральное государственное автономное образовательное учреждение высшего образования "Омский государственный технический университет" (ОмГТУ) Комбинированный электрод экг-метаболиты

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100179402A1 (en) * 2005-03-10 2010-07-15 Dexcom, Inc. System and methods for processing analyte sensor data for sensor calibration
US20140290054A1 (en) * 2013-03-27 2014-10-02 Google Inc. Systems and Methods for Encapsulating Electronics in a Mountable Device
US20140323819A1 (en) * 2013-04-29 2014-10-30 Elwha LLC, a limited liability company of the State of Delaware Multi-parameter test units for initial indication of medical symptoms
US20170102358A1 (en) * 2014-12-18 2017-04-13 Agilome, Inc. Chemically-sensitive field effect transistors, systems, and methods for manufacturing and using the same

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100179402A1 (en) * 2005-03-10 2010-07-15 Dexcom, Inc. System and methods for processing analyte sensor data for sensor calibration
US20140290054A1 (en) * 2013-03-27 2014-10-02 Google Inc. Systems and Methods for Encapsulating Electronics in a Mountable Device
US20140323819A1 (en) * 2013-04-29 2014-10-30 Elwha LLC, a limited liability company of the State of Delaware Multi-parameter test units for initial indication of medical symptoms
US20170102358A1 (en) * 2014-12-18 2017-04-13 Agilome, Inc. Chemically-sensitive field effect transistors, systems, and methods for manufacturing and using the same

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
KUMAR ET AL.: "Highly Sensitive Protein Functionalized Nanostructured Hafnium Oxide Based Biosensing Platform for Non-Invasive Oral Cancer Detection", SENSORS AND ACTUATORS B, vol. 235, 10 May 2016 (2016-05-10), pages 1 - 10, XP029660422, doi:10.1016/j.snb.2016.05.047 *
MANNOOR ET AL.: "Graphene-Based Wireless Bacteria Detection on Tooth Enamel", NATURE COMMUNICATIONS, vol. 3, no. 763, 27 March 2012 (2012-03-27), pages 1 - 8, XP055237293, doi:10.1038/ncomms1767 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20210293776A1 (en) * 2020-03-18 2021-09-23 Nanodx, Inc. Measurement systems and associated techniques for sensing electrical characteristics of a sensor
WO2022232373A1 (fr) * 2021-04-28 2022-11-03 Freire Marcelo Système et procédé de biodétection immunitaire et moléculaire par l'intermédiaire d'une brosse à dents
RU220817U1 (ru) * 2023-07-10 2023-10-04 Федеральное государственное автономное образовательное учреждение высшего образования "Омский государственный технический университет" (ОмГТУ) Комбинированный электрод экг-метаболиты

Similar Documents

Publication Publication Date Title
Li et al. Pattern recognition receptors in health and diseases
Walsh et al. Recommendations for childhood cancer screening and surveillance in DNA repair disorders
Gutiérrez-Sanz et al. Direct, label-free, and rapid transistor-based immunodetection in whole serum
Wang et al. Clinically relevant detection of Streptococcus pneumoniae with DNA-antibody nanostructures
CN106537130B (zh) 用于分析物检测的电化学传感器
Hasseb et al. Application of molecularly imprinted polymers for electrochemical detection of some important biomedical markers and pathogens
US9029168B2 (en) Use and making of biosensors utilizing antimicrobial peptides for highly sensitive biological monitoring
CN103069270B (zh) 通过使用半抗体生物传感器形成稳态环用于检测和调节血管内皮生长因子（vegf）的方法和装置
CN102279270B (zh) 利用聚集诱导发光监测β淀粉样蛋白聚集过程的方法
Lim et al. New sensors for extracellular vesicles: insights on constituent and associated biomarkers
WO2019173220A1 (fr) Système et procédé de détection sans fil d'état de santé buccale
Liu et al. On-chip structure-switching aptamer-modified magnetic nanobeads for the continuous monitoring of interferon-gamma ex vivo
JP2013178260A5 (fr)
CN102782501A (zh) 在无症状患者中基于il-6检测的全身炎性应答综合征和败血症的早期诊断和预测
Vásquez et al. Detection of COVID-19-related biomarkers by electrochemical biosensors and potential for diagnosis, prognosis, and prediction of the course of the disease in the context of personalized medicine
CA3075908A1 (fr) Dosages immunologiques pour la detection de proteines ran
Soares et al. Diagnostics of SARS-CoV-2 infection using electrical impedance spectroscopy with an immunosensor to detect the spike protein
US10925529B2 (en) System, method, and device for detecting postoperative complications
Costa et al. Molecularly imprinted polymer as a synthetic antibody for the biorecognition of hazelnut Cor a 14-allergen
Stefan-van Staden et al. Molecular recognition of IL-8, IL-10, IL-12, and IL-15 in biological fluids using phthalocyanine-based stochastic sensors
Oguzhan Caglayan Electrochemical aptasensors for early cancer diagnosis: a review
Liu et al. Electrochemical biosensors based on saliva electrolytes for rapid detection and diagnosis
Chavan et al. Recombinant histidine-tagged nano-protein-based highly sensitive electro-sensing device for salivary cortisol
Cardinell et al. Multi-biomarker detection following traumatic brain injury
Altıntaş et al. Exploring the versatility of exosomes: a review on isolation, characterization, detection methods, and diverse applications

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19763468

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19763468

Country of ref document: EP

Kind code of ref document: A1