CN109085223B - Preparation method of implantable biosensor - Google Patents
Preparation method of implantable biosensor Download PDFInfo
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- CN109085223B CN109085223B CN201810968649.0A CN201810968649A CN109085223B CN 109085223 B CN109085223 B CN 109085223B CN 201810968649 A CN201810968649 A CN 201810968649A CN 109085223 B CN109085223 B CN 109085223B
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- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 claims description 66
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 54
- 108010022355 Fibroins Proteins 0.000 claims description 47
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 claims description 44
- 239000000243 solution Substances 0.000 claims description 33
- 239000011787 zinc oxide Substances 0.000 claims description 33
- 239000010931 gold Substances 0.000 claims description 31
- 229910052737 gold Inorganic materials 0.000 claims description 31
- 238000004070 electrodeposition Methods 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 29
- 239000002073 nanorod Substances 0.000 claims description 28
- 229960000890 hydrocortisone Drugs 0.000 claims description 22
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 20
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 20
- 239000004366 Glucose oxidase Substances 0.000 claims description 20
- 239000007864 aqueous solution Substances 0.000 claims description 20
- 239000001110 calcium chloride Substances 0.000 claims description 20
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 20
- 229940116332 glucose oxidase Drugs 0.000 claims description 20
- ONDPHDOFVYQSGI-UHFFFAOYSA-N zinc nitrate Chemical compound [Zn+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ONDPHDOFVYQSGI-UHFFFAOYSA-N 0.000 claims description 20
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 19
- 238000000151 deposition Methods 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 239000000758 substrate Substances 0.000 claims description 13
- 229920001577 copolymer Polymers 0.000 claims description 12
- 230000008021 deposition Effects 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 12
- 239000008367 deionised water Substances 0.000 claims description 11
- 229910021641 deionized water Inorganic materials 0.000 claims description 11
- 239000003792 electrolyte Substances 0.000 claims description 11
- 108010015776 Glucose oxidase Proteins 0.000 claims description 10
- WQZGKKKJIJFFOK-DVKNGEFBSA-N alpha-D-glucose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-DVKNGEFBSA-N 0.000 claims description 10
- 239000011521 glass Substances 0.000 claims description 10
- 235000019420 glucose oxidase Nutrition 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims description 9
- 230000008569 process Effects 0.000 claims description 9
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 8
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 claims description 7
- 239000000427 antigen Substances 0.000 claims description 7
- 102000036639 antigens Human genes 0.000 claims description 7
- 108091007433 antigens Proteins 0.000 claims description 7
- 238000004140 cleaning Methods 0.000 claims description 7
- 238000000502 dialysis Methods 0.000 claims description 7
- 230000004048 modification Effects 0.000 claims description 7
- 238000012986 modification Methods 0.000 claims description 7
- ZNNZYHKDIALBAK-UHFFFAOYSA-M potassium thiocyanate Chemical compound [K+].[S-]C#N ZNNZYHKDIALBAK-UHFFFAOYSA-M 0.000 claims description 7
- 229940116357 potassium thiocyanate Drugs 0.000 claims description 7
- 238000010894 electron beam technology Methods 0.000 claims description 6
- 238000001556 precipitation Methods 0.000 claims description 6
- AASBXERNXVFUEJ-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) propanoate Chemical compound CCC(=O)ON1C(=O)CCC1=O AASBXERNXVFUEJ-UHFFFAOYSA-N 0.000 claims description 5
- 230000001112 coagulating effect Effects 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 5
- 125000004119 disulfanediyl group Chemical group *SS* 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 239000012456 homogeneous solution Substances 0.000 claims description 5
- 238000000465 moulding Methods 0.000 claims description 5
- 238000007790 scraping Methods 0.000 claims description 5
- 238000007711 solidification Methods 0.000 claims description 5
- 230000008023 solidification Effects 0.000 claims description 5
- 238000005303 weighing Methods 0.000 claims description 5
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 4
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 238000009835 boiling Methods 0.000 claims description 3
- XHFLOLLMZOTPSM-UHFFFAOYSA-M sodium;hydrogen carbonate;hydrate Chemical compound [OH-].[Na+].OC(O)=O XHFLOLLMZOTPSM-UHFFFAOYSA-M 0.000 claims description 3
- 238000007605 air drying Methods 0.000 claims description 2
- 238000005520 cutting process Methods 0.000 claims description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 238000012544 monitoring process Methods 0.000 abstract description 3
- 230000036541 health Effects 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 description 15
- 241000282414 Homo sapiens Species 0.000 description 8
- 239000000126 substance Substances 0.000 description 6
- 239000000523 sample Substances 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 239000004744 fabric Substances 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
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- 229910052751 metal Inorganic materials 0.000 description 3
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- 239000003381 stabilizer Substances 0.000 description 2
- 206010063385 Intellectualisation Diseases 0.000 description 1
- 238000005054 agglomeration Methods 0.000 description 1
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- 230000004075 alteration Effects 0.000 description 1
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- 239000003054 catalyst Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
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- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
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- 239000000284 extract Substances 0.000 description 1
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- 238000010438 heat treatment Methods 0.000 description 1
- -1 however Substances 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229910052761 rare earth metal Inorganic materials 0.000 description 1
- 150000002910 rare earth metals Chemical class 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000010897 surface acoustic wave method Methods 0.000 description 1
- 238000006557 surface reaction Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000005676 thermoelectric effect Effects 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
-
- 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
- A61B5/1486—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
- A61B5/14865—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 invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
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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
- G01N33/53—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
- G01N33/553—Metal or metal coated
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- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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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
- G01N33/66—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood sugars, e.g. galactose
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Abstract
The invention relates to the field of sensors and discloses a preparation method of an implantable biosensor. The sensor has good flexibility, and can be applied to the fields of medical biological monitoring, environmental and health monitoring and the like.
Description
Technical Field
The invention relates to the field of sensors, in particular to a preparation method of an implantable biosensor.
Background
The sensor is an important device which can be used for detecting various kinds of sensory information in the nature by human beings and converting different signals and the sensory information into digitalization and intellectualization. In the process of the rapid development of human civilization information technology, the sensor plays an increasingly important role, and is one of important means for human beings to explore unknown world and understand real world and observe surrounding information. The sensor is an adjustable component which can convert non-electrical signals such as electricity, light, temperature, chemical action and the like into electrical signals.
The piezoelectric properties of zinc oxide can be applied to surface acoustic wave devices, bulk acoustic wave devices, acousto-optic devices, and short-wave semiconductor diodes. When the zinc oxide is doped with transition metal or rare earth metal, the zinc oxide can show ferroelectric property, simultaneously, the zinc oxide also has thermoelectric effect and chemical sensing characteristic, and can be used for sensors and detectors.
Implantable medical electronics are increasingly in demand and are increasingly demanding in terms of their performance capabilities. The existing implantable medical electronic device can not meet the high requirements of long-term in vivo implantation on the volume, stability and biocompatibility of the implantable medical device, and the problems of heating, capacity reduction, internal degeneration and the like often occur in the practical use process of the implantable sensor. Once such a power source has reached its useful life, the patient has to undergo a secondary operation to remove it from the body, a process that places a significant burden on the patient's mind and economics. Therefore, there is an urgent need to develop a new power supply for supplying power to the implanted electronic device, so as to provide a feasible solution to the above problems.
Disclosure of Invention
In order to solve the above technical problems, the present invention provides a method for preparing an implantable biosensor. The preparation method comprises the steps of extracting silk fibroin in silk fabrics, purifying, blending and modifying the silk fibroin by using a polystyrene-maleic anhydride copolymer, taking the modified silk fibroin blend as a substrate, using a nanogold measuring electrode as a conductive electrode, depositing the silk fibroin blend on the substrate by using an electron beam cold precipitation evaporator, uniformly growing a zinc oxide nanorod on the substrate, and preparing the zinc oxide nanorod by adopting an electrochemical deposition method, wherein capture probes of the sensor are monoclonal glucose oxidase and a cortisol antibody which are respectively combined with a glucose oxidase molecule and a cortisol antigen molecule, and are respectively functionalized on the surface of zinc oxide to obtain the implantable biosensor, and the implantable biosensor has more excellent detection performance compared with a traditional sensor.
The specific technical scheme of the invention is as follows: a method of making an implantable biosensor, comprising the steps of:
(1) preparation of a bioelectrochemical substrate: immersing raw silk into sodium bicarbonate water solution, boiling for 30-60 minutes, and washing with deionized water; dissolving the obtained degummed silk in a calcium chloride/ethanol water solution system at the temperature of 80-100 ℃ under stirring; subsequently, the obtained homogeneous solution was filtered with a microporous filter, dialyzed with a dialysis tube, and freeze-dried at-40 to-80 ℃ for 24 to 36 hours to obtain a silk fibroin film; and dissolving the silk fibroin film in a hexafluoroisopropanol solution, and air-drying at room temperature for 12-18 hours to obtain the silk fibroin.
The silk fibroin is natural high molecular fiber extracted from silk, has good mechanical properties and physical and chemical properties such as good flexibility, tensile strength, air permeability, moisture permeability, slow release property and the like, is used as a biological substrate, can be automatically degraded and absorbed by organisms after a set task is finished, and avoids secondary treatment.
(2) Blending modification of the silk fibroin film: weighing polystyrene-maleic anhydride copolymer and silk fibroin according to the mass ratio of 7:80-100, dissolving the polystyrene-maleic anhydride copolymer and the silk fibroin in a potassium thiocyanate aqueous solution, adding 3-7wt% of sodium hydroxide solution, stirring and mixing at 80-100 ℃, cooling to room temperature, standing and defoaming at-35-40 ℃ for 4-6h, placing on a glass plate at room temperature, scraping a film, and placing the glass plate in a coagulating bath for solidification and molding to obtain the modified silk fibroin film.
The silk fibroin film has certain defects, such as compactness, no hole, poor support property and the like, and limits the application of the silk fibroin film.
(3) Preparing a nano gold conductive electrode: and depositing the nano-gold measuring electrode into the modified silk fibroin film by using an electron beam cold precipitation evaporator, and cutting to obtain the nano-gold conductive electrode.
The nano gold has the excellent characteristics of large specific surface area, high surface reaction activity, unique catalytic performance and the like. In these methods, in order to avoid agglomeration of nano gold particles, a polymeric surface reducing agent and a stabilizing agent are usually added in the preparation process for protection, and these protecting agents are adsorbed on the particle surfaces, so that direct contact between particles is avoided through electrostatic action or steric hindrance, and colloidal particles can stably exist in a solvent, however, surfactants and stabilizing agents are usually poor in conductivity, have a great influence on a catalyst, and even cause gold activity reduction and even loss of catalytic activity. The technical problems can be effectively solved by adopting the method of the invention.
(4) Preparing a zinc oxide nanorod array: and (2) adopting an electrochemical deposition method, adopting a two-electrode system, taking a zinc nitrate solution as an electrolyte, placing the electrolyte in a water bath kettle at a constant temperature of 70-80 ℃, respectively taking the cut zinc foil and the nano-gold conductive electrode as an anode and a cathode of the electrodeposition, connecting the anode and the cathode into a circuit for electrodeposition, taking out the nano-gold conductive electrode after the deposition is finished, cleaning and drying the nano-gold conductive electrode, and obtaining the zinc oxide nanorod array.
Electrodeposition refers to the process of electrochemical deposition of a metal or alloy from an aqueous, non-aqueous or molten salt of its compound. The ease of metal electrodeposition and the morphology of the deposit are related to the nature of the deposited metal and also depend on the composition of the electrolyte, pH, temperature, current density, and other factors.
(5) The biosensor is prepared by diluting 10-15mmol of dithio [ succinimidyl propionate ] in 3-5 μ L of dimethyl sulfoxide, incubating for 2-3 hours, functionalizing on the surface of a zinc oxide nanorod array, preparing 3-5 μ L of α -glucose oxidase antibody with the concentration of 10-15 μ g/mL, incubating for 15-20 minutes, fixing glucose oxidase molecules on α -glucose oxidase antibody, functionalizing on the surface of the zinc oxide nanorod array, preparing 3-5 μ L of cortisol antibody with the concentration of 10-15 μ g/mL, incubating for 15-20 minutes, fixing cortisol antigen molecules on the cortisol antibody, and functionalizing on the surface of the zinc oxide nanorod array to obtain the implantable biosensor.
The invention takes monoclonal glucose oxidase combined with glucose oxidase molecules and cortisol antibody combined with cortisol antigen molecules as capture probes. The capture probe refers to a type of labeled molecules used for indicating the properties or physical states of specific substances (such as nucleic acids, proteins, cell structures and the like) in molecular biology and biochemical experiments, in the invention, glucose oxidase and cortisol antibodies serving as the probes can detect glucose molecules and cortisol molecules in a human body, and the information is transmitted to an external intelligent device for information conversion so as to judge the concentration of glucose in the human body, so that the implantable biosensor can be used for monitoring the health of the human body.
Preferably, in step (1), the concentration of the aqueous sodium bicarbonate solution is 5 to 20 wt%.
Preferably, in the step (1), the molar ratio of the calcium chloride to the ethanol in the calcium chloride/ethanol aqueous solution system is 1:2, and the total molar concentration of the calcium chloride/ethanol aqueous solution system is 1.8-2 mol/L.
Preferably, in step (1), the size of the microporous filter is 0.22-0.24 μm, and the size of the dialysis tubing is 3.5k molecular weight cut-off.
Preferably, in the step (1), the concentration of the hexafluoroisopropanol solution is 1-2 g/mL.
Preferably, in the step (2), the concentration of the potassium thiocyanate aqueous solution is 35-40 wt%.
Preferably, in step (3), the nano-gold conductive measuring electrode has a cut size of 2 × 8mm, and an undeposited portion thereof after deposition has a size of 2 × 2 mm.
Preferably, in the step (4), the concentration of the zinc nitrate solution is 0.002-0.2M.
Preferably, in the step (4), the current is controlled to be 0.9-1.1mA during the electrochemical deposition process, and the electrochemical deposition is finished after 4-6h after the deposition is started.
Preferably, in the step (5), the concentration of the dimethyl sulfoxide is 5 to 6. mu.g/mL.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention takes the biological material as the substrate, extracts and purifies the silk fibroin in the raw silk, has good mechanical property and physical and chemical properties, such as good flexibility, tensile strength, air permeability, moisture permeability, slow release property and the like, and uses the polystyrene-maleic anhydride copolymer to carry out blending modification, so that the biocompatibility is improved, the film forming property is more excellent, and the silk fibroin can be automatically degraded and absorbed by organisms after the biological material is used as a biological substrate and a given task is finished, thereby avoiding secondary treatment.
2. The nano gold is used as a conductive electrode, and the zinc oxide nano rod is deposited by an electrochemical deposition method, so that the nano gold has excellent ferroelectric property and chemical sensing characteristic, and has wide prospect in the field of sensors.
3. The biosensor can be implanted into a human body, and has two capture probes which can capture glucose molecules and cortisol molecules respectively so as to detect the glucose concentration in the human body.
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1
(1) Preparation of a bioelectrochemical substrate:
extracting silk fibroin in silk fabric, immersing raw silk into 5wt% sodium bicarbonate water solution, boiling for 30 minutes, and thoroughly washing with 200-400mL deionized water. Dissolving the obtained degummed silk in a calcium chloride/ethanol aqueous solution system under the condition of vigorous stirring at 80 ℃, wherein the molar ratio of calcium chloride to ethanol in the calcium chloride/ethanol aqueous solution system is 1:2, and the total molar concentration of the calcium chloride/ethanol aqueous solution system is 1.8 mol/L. Subsequently, the obtained homogeneous solution was filtered with a microporous filter, dialyzed using a dialysis tube having a specification of 0.22 μm and a specification of 3.5k molecular weight cut-off, and further freeze-dried at-40 ℃ for 24 hours to obtain a silk fibroin film. Then, the silk fibroin film was dissolved in 1 g/mL hexafluoroisopropanol solution, placed in a hood, and air-dried at room temperature for 12 hours.
(2) Blending modification of the silk fibroin film:
weighing 7g of polystyrene-maleic anhydride copolymer and 90g of silk fibroin obtained in the step (1), dissolving the polystyrene-maleic anhydride copolymer and the silk fibroin in 38wt% of potassium thiocyanate aqueous solution, adding 5mL of 5% sodium hydroxide solution, stirring and mixing at 80 ℃, cooling to room temperature, standing and defoaming at minus 35 ℃ for 4h, scraping a film on a glass plate at room temperature, and then placing the glass plate in a coagulating bath for solidification and molding.
(3) Preparing a nano gold conductive electrode:
and (3) depositing a nanogold measuring electrode into the modified silk fibroin film in the step (2) by using an electron beam cold precipitation evaporator. The nano-gold conductive measuring electrode is 2 x 8mm in size after being cut, and is 2 x 2mm in size after being covered.
(4) Preparing a zinc oxide nano rod:
and (3) adopting an electrochemical deposition method, adopting a two-electrode system, taking a zinc nitrate solution as an electrolyte, placing the electrolyte in a water bath kettle at a constant temperature of 70 ℃, enabling the concentration of the zinc nitrate solution to be 0.002M, respectively connecting the cut zinc foil and the nano gold conductive electrode obtained in the step (3) as an anode and a cathode of the electrodeposition to be connected into a circuit for electrodeposition, enabling the size of the zinc foil to be 4 x 4mm, and repeatedly cleaning with deionized water and absolute ethyl alcohol by ultrasonic waves for 3 times. And controlling the current to be 0.9mA in the electrochemical deposition process, starting the deposition, finishing the electrochemical deposition after 4 hours, taking out the nano gold conductive electrode after the deposition is finished, and washing and drying the nano gold conductive electrode by using deionized water to obtain the zinc oxide nano rod array.
(5) Preparation of the biosensor:
diluting 10mmol of dithio [ succinimidyl propionate ] in 3 mu L of dimethyl sulfoxide, wherein the concentration of the dimethyl sulfoxide is 5 mu g/mL, incubating for 2 hours, functionalizing on the surface of a zinc oxide nanorod array, preparing 3 mu L of α -glucose oxidase antibody with the concentration of 10 mu g/mL, incubating for 15 minutes, fixing glucose oxidase molecules on a α -glucose oxidase antibody, functionalizing on the surface of the zinc oxide nanorod array, preparing 4 mu L of cortisol antibody with the concentration of 12 mu g/mL, incubating for 18 minutes, fixing cortisol antigen molecules on the cortisol antibody, and functionalizing on the surface of the zinc oxide nanorod array to obtain the implantable biosensor.
Example 2
(1) Preparation of a bioelectrochemical substrate:
silk fibroin was extracted from silk fabrics, and raw silk was immersed in a 15wt% aqueous sodium bicarbonate solution, boiled for 70 minutes, and thoroughly rinsed with 150mL of deionized water. Dissolving the obtained degummed silk in a calcium chloride/ethanol aqueous solution system under the condition of vigorous stirring at 100 ℃, wherein the molar ratio of calcium chloride to ethanol in the calcium chloride/ethanol aqueous solution system is 1:2, and the total molar concentration of the calcium chloride/ethanol aqueous solution system is 2 mol/L. Subsequently, the obtained homogeneous solution was filtered with a microporous filter, dialyzed using a dialysis tube having a specification of 0.23 μm and a specification of 3.5k molecular weight cut-off, and further freeze-dried at-80 ℃ for 30 hours to obtain a silk fibroin film. Then, a certain amount of the silk fibroin film was dissolved in hexafluoroisopropanol solution at a concentration of 1 g/mL, placed in a hood, and air-dried at room temperature for 16 hours.
(2) Blending modification of the silk fibroin film:
weighing 7g of polystyrene-maleic anhydride copolymer and 90g of silk fibroin obtained in the step (1), dissolving the polystyrene-maleic anhydride copolymer and the silk fibroin in 38% potassium thiocyanate aqueous solution, adding 8mL of 5% sodium hydroxide solution, stirring and mixing at 90 ℃, cooling to room temperature, standing and defoaming at minus 40 ℃ for 4h, scraping a film on a glass plate at room temperature, and then placing the glass plate in a coagulating bath for solidification and molding.
(3) Preparing a nano gold conductive electrode:
and (3) depositing a nanogold measuring electrode into the modified silk fibroin film in the step (2) by using an electron beam cold precipitation evaporator. The nano-gold conductive measuring electrode is 2 x 8mm in size after being cut, and is 2 x 2mm in size after being covered.
(4) Preparing a zinc oxide nano rod:
and (3) adopting an electrochemical deposition method, adopting a two-electrode system, taking a zinc nitrate solution as an electrolyte, placing the electrolyte in a water bath kettle at a constant temperature of 70 ℃, enabling the concentration of the zinc nitrate solution to be 0.008M, respectively taking the cut zinc foil and the nano gold conductive electrode obtained in the step (3) as an anode and a cathode of the electrodeposition to be connected into a circuit for electrodeposition, enabling the size of the zinc foil to be 4mm, and repeatedly cleaning with deionized water and absolute ethyl alcohol for 3 times by ultrasonic. And controlling the current to be 1.1mA in the electrochemical deposition process, starting the deposition, finishing the electrochemical deposition after 4 hours, taking out the nano gold conductive electrode after the deposition is finished, and cleaning and drying the nano gold conductive electrode by using deionized water to obtain the zinc oxide nano rod array.
(5) Preparation of the biosensor:
diluting 10mmol of dithio [ succinimidyl propionate ] in 3 mu L of dimethyl sulfoxide, wherein the concentration of the dimethyl sulfoxide is 5 mu g/mL, incubating for 2 hours, functionalizing on the surface of a zinc oxide nanorod array, preparing 3 mu L of α -glucose oxidase antibody with the concentration of 15 mu g/mL, incubating for 17 minutes, fixing glucose oxidase molecules on a α -glucose oxidase antibody, functionalizing on the surface of the zinc oxide nanorod array, preparing 3 mu L of cortisol antibody with the concentration of 15 mu g/mL, incubating for 15 minutes, fixing cortisol antigen molecules on the cortisol antibody, and functionalizing on the surface of the zinc oxide nanorod array to obtain the implantable biosensor.
Example 3
(1) Preparation of a bioelectrochemical substrate:
silk fibroin was extracted from silk fabrics, and raw silk was immersed in a 20wt% aqueous sodium bicarbonate solution, boiled for 60 minutes, and thoroughly rinsed with 400mL of deionized water. Dissolving the obtained degummed silk in a calcium chloride/ethanol aqueous solution system under the condition of vigorous stirring at 100 ℃, wherein the molar ratio of calcium chloride to ethanol in the calcium chloride/ethanol aqueous solution system is 1:2, and the total molar concentration of the calcium chloride/ethanol aqueous solution system is 2 mol/L. Subsequently, the obtained homogeneous solution was filtered with a microporous filter, dialyzed using a dialysis tube having a specification of 0.24 μm and a specification of 3.5k molecular weight cut-off, and further freeze-dried at-80 ℃ for 36 hours to obtain a silk fibroin film. Then, a certain amount of the silk fibroin film was dissolved in hexafluoroisopropanol solution having a concentration of 2 g/mL, placed in a hood, and air-dried at room temperature for 18 hours.
(2) Blending modification of the silk fibroin film:
weighing 7g of polystyrene-maleic anhydride copolymer and 90g of silk fibroin obtained in the step (1), dissolving the polystyrene-maleic anhydride copolymer and the silk fibroin in 38% potassium thiocyanate aqueous solution, adding 10mL of 5% sodium hydroxide solution, stirring and mixing at 100 ℃, cooling to room temperature, standing and defoaming at minus 38 ℃ for 5 hours, scraping a film on a glass plate at room temperature, and then placing the glass plate in a coagulating bath for solidification and molding.
(3) Preparing a nano gold conductive electrode:
and (3) depositing a nanogold measuring electrode into the modified silk fibroin film in the step (2) by using an electron beam cold precipitation evaporator. The nano-gold conductive measuring electrode is 2 x 8mm in size after being cut, and is 2 x 2mm in size after being covered.
(4) Preparing a zinc oxide nano rod:
and (3) adopting an electrochemical deposition method, adopting a two-electrode system, taking a zinc nitrate solution as an electrolyte, placing the electrolyte in a water bath kettle at a constant temperature of 80 ℃, enabling the concentration of the zinc nitrate solution to be 0.1M, respectively connecting the cut zinc foil and the nano gold conductive electrode obtained in the step (3) as an anode and a cathode of the electrodeposition to be connected into a circuit for electrodeposition, enabling the size of the zinc foil to be 4mm, and repeatedly cleaning the zinc foil for 3 times by using deionized water and absolute ethyl alcohol in an ultrasonic mode. And controlling the current to be 0.9mA in the electrochemical deposition process, starting the deposition, finishing the electrochemical deposition after 6 hours, taking out the nano gold conductive electrode after the deposition is finished, and cleaning and drying the nano gold conductive electrode by using deionized water to obtain the zinc oxide nano rod array.
(5) Preparation of the biosensor:
diluting 15mmol of dithio [ succinimidyl propionate ] in 5 mu L of dimethyl sulfoxide, wherein the concentration of the dimethyl sulfoxide is 6 mu g/mL, incubating for 3 hours, functionalizing on the surface of a zinc oxide nanorod array, preparing 5 mu L of α -glucose oxidase antibody with the concentration of 15 mu g/mL, incubating for 20 minutes, fixing glucose oxidase molecules on a α -glucose oxidase antibody, functionalizing on the surface of the zinc oxide nanorod array, preparing 5 mu L of cortisol antibody with the concentration of 10 mu g/mL, incubating for 20 minutes, fixing cortisol antigen molecules on the cortisol antibody, and functionalizing on the surface of the zinc oxide nanorod array to obtain the implantable biosensor.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (10)
1. A method of making an implantable biosensor, comprising the steps of:
(1) preparation of a bioelectrochemical substrate: immersing raw silk into sodium bicarbonate water solution, boiling for 30-60 minutes, and washing with deionized water; dissolving the obtained degummed silk in a calcium chloride/ethanol water solution system at the temperature of 80-100 ℃ under stirring; subsequently, the obtained homogeneous solution was filtered with a microporous filter, dialyzed with a dialysis tube, and freeze-dried at-40 to-80 ℃ for 24 to 36 hours to obtain a silk fibroin film; dissolving the silk fibroin film in a hexafluoroisopropanol solution, and air-drying at room temperature for 12-18 hours to obtain silk fibroin;
(2) blending modification of the silk fibroin film: weighing polystyrene-maleic anhydride copolymer and silk fibroin according to a mass ratio of 7:80-100, dissolving the polystyrene-maleic anhydride copolymer and the silk fibroin in a potassium thiocyanate aqueous solution, adding 3-7wt% of sodium hydroxide solution, stirring and mixing at 80-100 ℃, cooling to room temperature, standing and defoaming at-35-40 ℃ for 4-6h, placing on a glass plate at room temperature, scraping a film, and placing the glass plate in a coagulating bath for solidification and molding to obtain a modified silk fibroin film;
(3) preparing a nano gold conductive electrode: the nanogold measuring electrode uses an electron beam cold precipitation evaporator to deposit nanogold into the modified silk fibroin film substrate, and the deposited silk fibroin film substrate is cut to obtain a nanogold conducting electrode;
(4) preparing a zinc oxide nanorod array: using an electrochemical deposition method, adopting a two-electrode system, taking a zinc nitrate solution as an electrolyte, placing the electrolyte in a water bath kettle at a constant temperature of 70-80 ℃, respectively taking the cut zinc foil and the nano-gold conductive electrode as an anode and a cathode of electrodeposition, connecting the anode and the cathode into a circuit for electrodeposition, and after the deposition is finished, taking the nano-gold conductive electrode out, cleaning and drying to obtain a zinc oxide nano-rod array;
(5) the biosensor is prepared by diluting 10-15mmol of dithio [ succinimidyl propionate ] in 3-5 mu L of dimethyl sulfoxide, incubating for 2-3 hours, functionalizing on the surface of a zinc oxide nanorod array, preparing 3-5 mu L of α -glucose oxidase antibody with the concentration of 10-15 mu g/mL, incubating for 15-20 minutes, fixing glucose oxidase molecules on α -glucose oxidase antibody, further fixing and modifying on the surface of the zinc oxide nanorod array, preparing 3-5 mu L of cortisol antibody with the concentration of 10-15 mu g/mL, incubating for 15-20 minutes, fixing cortisol antigen molecules on the cortisol antibody, further fixing and modifying on the surface of the zinc oxide nanorod array, and obtaining the implantable biosensor.
2. The method of claim 1, wherein the concentration of the aqueous sodium bicarbonate solution in step (1) is 5 to 20 wt%.
3. The method of claim 1, wherein in step (1), the molar ratio of calcium chloride to ethanol in the calcium chloride/ethanol aqueous solution system is 1:2, and the total molar concentration of the calcium chloride/ethanol aqueous solution system is 1.8-2 mol/L.
4. The method of claim 1, wherein in step (1), the microporous filter has a size of 0.22-0.24 μm and the dialysis tubing has a size of 3.5k molecular weight cut-off.
5. The method of claim 1, wherein in step (1), the hexafluoroisopropanol solution is present at a concentration of 1-2 g/mL.
6. The method of claim 1, wherein in step (2), the concentration of the aqueous solution of potassium thiocyanate is 35-40 wt%.
7. The method according to claim 1, wherein in step (3), the nano-gold conductive electrode obtained after cutting has a size of 2 x 8mm, and the non-deposited part thereof after deposition has a size of 2 x 2 mm.
8. The method of claim 1, wherein in the step (4), the concentration of the zinc nitrate solution is 0.002-0.2M.
9. The method of claim 1, wherein in the step (4), the current is controlled to be 0.9 to 1.1mA during the electrochemical deposition process, and the electrochemical deposition is finished 4 to 6 hours after the deposition is started.
10. The method of claim 1, wherein the concentration of the dimethylsulfoxide in step (5) is 5 to 6 μ g/mL.
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Non-Patent Citations (4)
| Title |
|---|
| Conducting polymer-silk biocomposites for flexible and biodegradable electrochemical sensors;Ramendra K.Pal et.al;《Biosensors and Bioelectronics》;20160308;第81卷;第294-302页 * |
| Development of a disposable potentiometric antibody immobilized ZnO nanotubes based sensor for the detection of C-reactive protein;Z.H. Ibupoto et.al;《Sensors and Actuators B: Chemical》;20120406;第166卷;第809-814页 * |
| Lancet-free and label-free diagnostics of glucose in sweat using Zinc Oxide based flexible bioelectronics;Rujuta D. Munje et.al;《Sensors and Actuators B: Chemical》;20160718;第238卷;第482-490页 * |
| 丝素蛋白材料生物降解行为的研究;徐亚梅;《中国优秀硕士学位论文全文数据库（电子期刊） 医药卫生科技辑》;20120615(第06期);第10-15页 * |
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