CN115266870A - Preparation method of wearable micro-fluidic quasi-solid biochip based on gel, product and application thereof - Google Patents
Preparation method of wearable micro-fluidic quasi-solid biochip based on gel, product and application thereof Download PDFInfo
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- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 claims abstract description 14
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229940116269 uric acid Drugs 0.000 claims abstract description 14
- 239000000017 hydrogel Substances 0.000 claims abstract description 13
- 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 claims abstract description 12
- 239000008103 glucose Substances 0.000 claims abstract description 12
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims description 12
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- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 3
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 230000003197 catalytic effect Effects 0.000 abstract description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 12
- 238000012360 testing method Methods 0.000 description 10
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- 239000003575 carbonaceous material Substances 0.000 description 5
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- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- -1 polyethylene terephthalate Polymers 0.000 description 2
- 229920000139 polyethylene terephthalate Polymers 0.000 description 2
- 239000005020 polyethylene terephthalate Substances 0.000 description 2
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- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical group [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003592 biomimetic effect Effects 0.000 description 1
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- 238000000970 chrono-amperometry Methods 0.000 description 1
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- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000840 electrochemical analysis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910021389 graphene Inorganic materials 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- DCYOBGZUOMKFPA-UHFFFAOYSA-N iron(2+);iron(3+);octadecacyanide Chemical compound [Fe+2].[Fe+2].[Fe+2].[Fe+3].[Fe+3].[Fe+3].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] DCYOBGZUOMKFPA-UHFFFAOYSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000010339 medical test Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229960003351 prussian blue Drugs 0.000 description 1
- 239000013225 prussian blue Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
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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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- 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
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- 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
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- 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
- B01L2300/12—Specific details about materials
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- Chemical Kinetics & Catalysis (AREA)
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Abstract
The invention discloses a preparation method of a wearable micro-fluidic quasi-solid biochip based on gel, a product and application thereof, wherein the biochip is prepared by printing a conductive hydrogel containing a nano bionic enzyme material with good conductivity on a flexible substrate after adjusting the viscosity; the chip construction method is simple, strong in operability and low in cost. The conductive gel in the chip contains the nano bionic enzyme material for specifically detecting the biomolecules, so that the micro-fluidic chip has good catalytic performance on the specific biomolecules, and the micro-channel containing different nano bionic enzyme materials can simultaneously carry out real-time high-sensitivity monitoring on various biomolecules in human sweat; the flexible wearable micro-fluidic quasi-solid biochip constructed by the full printing technology has good swelling property and water absorption property, and can quickly absorb human sweat to realize the simultaneous real-time high-sensitivity electrochemical monitoring of various biomolecules such as glucose, uric acid, lactic acid, amino acid, urea and the like in the sweat.
Description
Technical Field
The invention relates to the field of biological detection, in particular to a wearable micro-fluidic quasi-solid biochip based on gel, and also relates to a preparation method and application of the wearable micro-fluidic quasi-solid biochip.
Background
The point-of-care testing (POCT) technology is a novel testing technology with great potential in the field of modern medical testing. Compared with the traditional clinical examination, the method has the advantages of rapid detection, simple and convenient operation, low cost, small sample consumption and the like. With the wide application of various biosensing technologies such as screen printing electrodes, immunolabeling, enzyme-linked technology, biochip technology and the like in POCT, compared with the traditional biochemical analyzer, the analysis time is greatly shortened, the detection cost is reduced, and the convenience for monitoring the patient at any time is provided for the patient. At present, main development companies include Roche, germany Bayer, yapeh, japan Kyoto, and the like. The main products with large market capacity are blood sugar test strips and uric acid test strips, but most of the commercially available test strips for detecting blood sugar, uric acid and the like are manufactured by using an enzyme biosensor and a screen printing technology. The enzyme has high cost, complex fixation and poor stability, and the activity of the enzyme is easily influenced by factors such as pH, temperature, humidity, toxic chemicals and the like, so that the commercially available test strip has high cost and short shelf life, and is usually stored in a refrigerator at 4 ℃, particularly the shelf life of the test strip after unsealing is only three months. In addition, such test strips require a patient to perform a needle blood sampling test and cannot achieve continuous dynamic monitoring, which limits the application of the test strips in daily life.
Therefore, aiming at the problems and challenges existing in the practical application of the current enzyme electrochemical test strip, a new generation of bionic enzyme electrochemical sensing chip with high performance, low cost and non-needle blood sampling is urgently needed to be developed.
Disclosure of Invention
In view of the above, an object of the present invention is to provide a method for preparing a gel-based wearable microfluidic quasi-solid biochip; the second purpose of the invention is to provide a wearable micro-fluidic quasi-solid biochip prepared by the method; the invention also aims to provide application of the wearable micro-fluidic quasi-solid biochip in detection of sweat biomolecules discharged from a human body.
In order to achieve the purpose, the invention provides the following technical scheme:
1. the preparation method of the wearable micro-fluidic quasi-solid biochip based on the gel comprises the following specific steps: and (3) adjusting the viscosity of the conductive hydrogel containing the nano bionic enzyme material with good conductivity, printing the conductive hydrogel on a flexible substrate to prepare a working electrode, and then obtaining the biochip together with a counter electrode.
Preferably, the nano bionic enzyme material is bionic enzyme for detecting biomolecules in sweat discharged from a human body.
In the invention, the biomolecule in the sweat discharged from the human body is glucose, uric acid, lactic acid, amino acid or urea.
Preferably, the conductive hydrogel is a hydrogel made of gelatin.
Preferably, the concentration of the nano bionic enzyme material with good conductivity is 1-10mg/mL.
Preferably, the flexible substrate is a PET film or a PE film.
2. The wearable micro-fluidic quasi-solid biochip prepared by the preparation method of the gel-based wearable micro-fluidic quasi-solid biochip.
3. The wearable micro-fluidic quasi-solid biochip is applied to detecting the sweat biomolecules discharged from a human body.
Preferably, the biomolecule in sweat discharged from the human body is glucose, uric acid, lactic acid, amino acid or urea.
The invention has the beneficial effects that: the invention develops novel conductive gel based on hydrogel with good biocompatibility and nano bionic enzyme material with good conductivity, and constructs a wearable micro-fluidic quasi-solid biochip on a flexible substrate by a full printing technology. Different microchannels of the wearable micro-fluidic quasi-solid biochip contain different nano bionic enzyme materials, and the wearable micro-fluidic quasi-solid biochip has good swelling property and water absorption property, so that the wearable micro-fluidic quasi-solid biochip can be applied to skin to realize real-time high-sensitivity electrochemical monitoring of various biomolecules such as glucose, uric acid, lactic acid, amino acid and urea in human sweat by rapidly absorbing the human sweat. The wearable micro-fluidic chip based on the conductive gel has high sensitivity, strong specificity and good stability, can be stored for 1 to 2 years under the room temperature non-sealing condition, still keeps higher sensitivity, and can greatly reduce the production cost. In addition, the wearable microfluidic quasi-solid biochip can effectively avoid needle blood sampling and infection risks brought by the needle blood sampling, has stronger practicability and applicability to daily detection of patients, and highlights potential application of the wearable microfluidic quasi-solid biochip in clinical diagnosis and daily disease monitoring.
Drawings
In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
fig. 1 is a schematic diagram of a wearable micro-fluidic quasi-solid biochip modified with different biomimetic enzyme nanomaterials (the central black point of the diagram is a counter electrode, and the peripheral black points are working electrodes).
FIG. 2 shows the results of biomolecule detection (a: glucose, b: uric acid, c: cholesterol, d: chip structure).
Detailed Description
The present invention is further described with reference to the following drawings and specific examples so that those skilled in the art can better understand the present invention and can practice the present invention, but the examples are not intended to limit the present invention.
Example 1 construction of a gel-based wearable microfluidic quasi-solid biochip
The preparation method of the wearable micro-fluidic quasi-solid biochip based on the gel comprises the following specific steps:
(1) Materials with good biocompatibility such as gelatin and the like are prepared into hydrogel;
(2) Adding a nano bionic enzyme material with good conductivity into the prepared hydrogel to enable the final concentration of the nano bionic enzyme to be 1-10mg/mL to form conductive gel, adjusting the viscosity, then constructing on a flexible substrate such as a PET (polyethylene terephthalate) film or a PE (polyethylene) film by printing and other technologies) to obtain a working electrode, and then obtaining the wearable micro-fluidic quasi-solid biochip together with a counter electrode.
Further carrying out freeze drying on the mixture to obtain the wearable micro-fluidic quasi-solid biochip, wherein the structure is shown in figure 1.
Respectively containing different nanometer bionic enzyme materials in different microchannels, wherein the nanometer bionic enzyme material of glucose is selected from copper nanometer flower composite carbon material or platinum nanometer flower composite carbon material, etc.; the nano bionic enzyme material of the uric acid is selected from a Prussian blue composite nitrogen-doped carbon material, a monoatomic cobalt-modified nitrogen-doped carbon material and the like; the nano bionic enzyme material of cholesterol selects nano gold composite carbon nano material to wrap cholesterol enzyme.
Wherein the carbon material is but not limited to carbon nano-materials with good conductivity such as graphene, carbon nano-tubes, mxene, biochar and the like);
the shape and size, the channel thickness, the number of microelectrodes and the like of the microfluidic chip in the embodiment can be designed according to the requirements of specific experiments.
Example 2 application of gel-based wearable microfluidic quasi-solid biochip in biomolecule detection
(1) The prepared flexible wearable micro-fluidic chip is pasted on a position of a human body, which is easy to sweat;
(2) Glucose, uric acid and cholesterol in sweat discharged by diabetics and gout patients are treated by a chronoamperometry.
The results are shown in FIG. 2. The results showed that glucose, uric acid and cholesterol in sweat could be detected, and the lowest detected concentration of glucose was 5mM, the lowest detected concentration of uric acid was 0.3mM and the lowest detected concentration of cholesterol was 3mM.
According to the same principle, different nano bionic enzyme materials are used for detecting other biomolecules such as lactic acid, amino acid, urea and the like.
In conclusion, the wearable micro-fluidic quasi-solid biochip based on the gel can be obtained by the construction method 1, and the construction method is simple, strong in operability and low in cost. In addition, the conductive gel contains a nano bionic enzyme material for specifically detecting biomolecules, so that the micro-fluidic chip has good catalytic performance on specific biomolecules, and the micro-channel containing different nano bionic enzyme materials can simultaneously carry out real-time high-sensitivity monitoring on various biomolecules in human sweat. Compared with the traditional test strip, the novel conductive gel developed by combining the hydrogel with good biocompatibility and the nano bionic enzyme material with good conductivity has the advantages of good specificity, high sensitivity, easiness in storage and low cost. The flexible wearable micro-fluidic quasi-solid biochip constructed by the full printing technology has good swelling property and water absorption property, and can quickly absorb human sweat to realize the simultaneous real-time high-sensitivity electrochemical monitoring of various biomolecules such as glucose, uric acid, lactic acid, amino acid, urea and the like in the sweat.
The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitutions or changes made by the person skilled in the art on the basis of the present invention are all within the protection scope of the present invention. The protection scope of the invention is subject to the claims.
Claims (9)
1. The preparation method of the gel-based wearable micro-fluidic quasi-solid biochip is characterized by comprising the following specific steps: and (3) adjusting the viscosity of the conductive hydrogel containing the nano bionic enzyme material with good conductivity, printing the conductive hydrogel on a flexible substrate to prepare a working electrode, and then obtaining the biochip together with a counter electrode.
2. The method of preparing a wearable microfluidic quasi-solid biochip based on gel according to claim 1, wherein the method comprises the steps of: the nano bionic enzyme material is bionic enzyme for detecting biomolecules in sweat discharged by a human body.
3. The method of preparing a wearable microfluidic quasi-solid biochip based on gel according to claim 2, wherein the method comprises the steps of: the biomolecule in sweat discharged by human body is glucose, uric acid, lactic acid, amino acid or urea.
4. The method for preparing a wearable micro-fluidic quasi-solid biochip based on gel according to claim 2, characterized in that: the conductive hydrogel is hydrogel prepared from gelatin.
5. The method of preparing a wearable microfluidic quasi-solid biochip based on gel according to claim 2, wherein the method comprises the steps of: the concentration of the nano bionic enzyme material with good conductivity is 1-10mg/mL.
6. The method for preparing a wearable micro-fluidic quasi-solid biochip based on gel according to claim 2, characterized in that: the flexible substrate is a PET film or a PE film.
7. A wearable microfluidic quasi-solid biochip prepared by the method for preparing a wearable microfluidic quasi-solid biochip based on gel according to any one of claims 1 to 6.
8. Use of the wearable microfluidic quasi-solid biochip of claim 7 for detecting sweat biomolecules in a human body.
9. Use according to claim 8, characterized in that: the biological molecules in the sweat discharged by the human body are glucose, uric acid, lactic acid, amino acid or urea.
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| CN111855778A (en) * | 2020-07-14 | 2020-10-30 | 天津大学 | Electrochemical biosensor based on zwitter-ion hydrogel porous enzyme membrane modification |
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2021
- 2021-04-30 CN CN202110486443.6A patent/CN115266870A/en active Pending
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| CN104474980A (en) * | 2014-11-07 | 2015-04-01 | 中南大学 | Supermolecule hybridized hydrogel, graphene aerogel, preparation methods and application of two |
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