WO2021159074A1 - Graphene-based sensor for detecting hemoglobin in a biological sample - Google Patents

Graphene-based sensor for detecting hemoglobin in a biological sample Download PDF

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Publication number
WO2021159074A1
WO2021159074A1 PCT/US2021/017082 US2021017082W WO2021159074A1 WO 2021159074 A1 WO2021159074 A1 WO 2021159074A1 US 2021017082 W US2021017082 W US 2021017082W WO 2021159074 A1 WO2021159074 A1 WO 2021159074A1
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WIPO (PCT)
Prior art keywords
graphene layer
sample
sensor
hemoglobin
functionalized
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Ceased
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PCT/US2021/017082
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English (en)
French (fr)
Inventor
Namal NAWANA
Mehdi ABEDI
Reza MOLLAAGHABABA
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GrapheneDx Inc
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GrapheneDx Inc
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Priority to EP21708890.5A priority Critical patent/EP4100739A1/en
Priority to AU2021216478A priority patent/AU2021216478A1/en
Priority to CN202180024705.5A priority patent/CN115917318A/zh
Priority to CA3170353A priority patent/CA3170353A1/en
Priority to KR1020227030390A priority patent/KR20220136407A/ko
Priority to JP2022548190A priority patent/JP2023513543A/ja
Publication of WO2021159074A1 publication Critical patent/WO2021159074A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
    • G01N33/721—Haemoglobin
    • G01N33/726—Devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • G01N33/5438—Electrodes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Rigid containers without fluid transport within
    • B01L3/5082—Test tubes per se
    • B01L3/50825—Closing or opening means, corks, bungs
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
    • G01N33/545—Synthetic resin
    • G01N33/547—Synthetic resin with antigen or antibody attached to the carrier via a bridging agent
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57535—Immunoassay; Biospecific binding assay; Materials therefor for cancer of the large intestine, e.g. colon, rectum or anus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/5758—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/5758—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites
    • G01N33/57585—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites involving compounds identifiable in body fluids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
    • G01N33/721—Haemoglobin
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/16—Reagents, handling or storing thereof
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Rigid containers without fluid transport within
    • B01L3/5085—Rigid containers without fluid transport within for multiple samples, e.g. microtitration plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/795—Porphyrin- or corrin-ring-containing peptides
    • G01N2333/805—Haemoglobins; Myoglobins

Definitions

  • a method of detecting hemoglobin in a biological sample includes applying the sample to a graphene layer that is functionalized with a plurality of anti-hemoglobin antibodies (e.g., anti-human hemoglobin antibodies) and/or aptamers, measuring at least one electrical property of the functionalized graphene layer, and using the measured electrical property to determine whether hemoglobin is present in the sample.
  • the measured electrical property of the graphene layer can be, for example, electron mobility, electrical impedance (e.g., DC or AC electrical resistance or both), and electrical capacitance.
  • V 0uti _GLO the corresponding voltage
  • an analyzer 600 can be in communication with the voltage measuring circuitry 301 to receive the applied current and the measured voltage value and use these values to calculate the resistance of the antibody- functionalized graphene layer. The analyzer 600 can then employ the calculated resistance, e.g., a change in the resistance in response to exposure of the antibody-functionalized graphene layer to a sample under investigation, to determine, in accordance with the present teachings, whether the sample contains hemoglobin protein.
  • the sensor 700 can include electrically conductive pads 722a, 722b, 724a and 724b, similar to the embodiment shown in FIG. 2.
  • the sensor 700 further includes a reference electrode (G) 705 that is disposed on the same substrate 702 as that on which the graphene layer 701 is disposed (in other words, the reference electrode 705 is in substantially same plane as the graphene layer 701).
  • the reference electrode 705 can substantially surround the graphene layer 701.
  • the reference electrode 705 can be electrically connected to additional conductive pads 726 and 728 to allow application of an AC voltage as well as a DC ramp voltage to the reference electrode 705, e.g., in a manner discussed above.
  • the high sensitivity of a graphene-based sensor according to the present teachings e.g., characterized by an LOD of better than 10 pg-Hb/g ⁇ feces, for detecting hemoglobin protein in a stool sample can allow using only hemoglobin protein as a biomarker for the detection of colorectal cancer and pre-malignant colorectal neoplasia.
  • a sensor 1200 can include at least two sensing units 1210 and 1220, wherein the sensing unit 1210 is configured as a graphene-based sensor according to the present teachings to detect the presence of hemoglobin in a sample, such as a fecal sample.
  • the sensor unit 1220 is a graphene- based sensor that is configured to detect one or more mutations (e.g., deletions or additions) in the generic material(s) present in the sample under investigation.
  • the graphene-based sensing unit 1220 includes a graphene layer 1201 that is disposed over an S1O2 layer, which is in turn disposed on an underlying silicon layer 1202.
  • the graphene layer 1201 is functionalized with a catalytically deactivated Cas9 (dCas9) CRISPR complex 1203 (e.g., Cas9 (dCas9) CRISPR complex).
  • dCas9 CRISPR complex 1203 e.g., Cas9 (dCas9) CRISPR complex
  • a single measurement circuit 1240 is provided, which is coupled via a multiplexer 1250, to the sensing units for monitoring the voltages across the sensing units 1210/1220 in different time intervals (e.g., during successive time intervals).

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)
  • Inorganic Chemistry (AREA)
  • Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Peptides Or Proteins (AREA)
PCT/US2021/017082 2020-02-06 2021-02-08 Graphene-based sensor for detecting hemoglobin in a biological sample Ceased WO2021159074A1 (en)

Priority Applications (6)

Application Number Priority Date Filing Date Title
EP21708890.5A EP4100739A1 (en) 2020-02-06 2021-02-08 Graphene-based sensor for detecting hemoglobin in a biological sample
AU2021216478A AU2021216478A1 (en) 2020-02-06 2021-02-08 Graphene-based sensor for detecting hemoglobin in a biological sample
CN202180024705.5A CN115917318A (zh) 2020-02-06 2021-02-08 用于检测生物样品中的血红蛋白的石墨烯基传感器
CA3170353A CA3170353A1 (en) 2020-02-06 2021-02-08 Graphene-based sensor for detecting hemoglobin in a biological sample
KR1020227030390A KR20220136407A (ko) 2020-02-06 2021-02-08 생물학적 샘플 내 헤모글로빈을 검출하기 위한 그래핀-기반 센서
JP2022548190A JP2023513543A (ja) 2020-02-06 2021-02-08 生体試料中のヘモグロビンを検出するためのグラフェンベースのセンサ

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202062970919P 2020-02-06 2020-02-06
US62/970,919 2020-02-06

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WO2021159074A1 true WO2021159074A1 (en) 2021-08-12

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PCT/US2021/017082 Ceased WO2021159074A1 (en) 2020-02-06 2021-02-08 Graphene-based sensor for detecting hemoglobin in a biological sample

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US (1) US20210247409A1 (https=)
EP (1) EP4100739A1 (https=)
JP (1) JP2023513543A (https=)
KR (1) KR20220136407A (https=)
CN (1) CN115917318A (https=)
AU (1) AU2021216478A1 (https=)
CA (1) CA3170353A1 (https=)
WO (1) WO2021159074A1 (https=)

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US12546746B2 (en) * 2021-05-05 2026-02-10 Ekidna Sensing Inc. Systems and devices for estimating substance concentration
US20230182133A1 (en) * 2021-12-01 2023-06-15 Graphene-Dx. Inc. Systems and methods for processing and testing biological samples
WO2025024598A1 (en) * 2023-07-24 2025-01-30 The Regents Of The University Of California Biomarker detection using graphene field effect transistor sensor arrays and multichannel data acquisition

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KR20220136407A (ko) 2022-10-07
EP4100739A1 (en) 2022-12-14
CA3170353A1 (en) 2021-08-12
US20210247409A1 (en) 2021-08-12
JP2023513543A (ja) 2023-03-31
AU2021216478A1 (en) 2022-09-08
CN115917318A (zh) 2023-04-04

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