CN108627559B - Immunoassay method based on the Prussian blue self energizing electrochemical sensor building of aluminium- - Google Patents
Immunoassay method based on the Prussian blue self energizing electrochemical sensor building of aluminium- Download PDFInfo
- Publication number
- CN108627559B CN108627559B CN201810438328.XA CN201810438328A CN108627559B CN 108627559 B CN108627559 B CN 108627559B CN 201810438328 A CN201810438328 A CN 201810438328A CN 108627559 B CN108627559 B CN 108627559B
- Authority
- CN
- China
- Prior art keywords
- prussian blue
- aluminium
- electrode
- electrochemical sensor
- self energizing
- Prior art date
- Legal status (The legal status 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 status listed.)
- Expired - Fee Related
Links
Classifications
-
- 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
- G01N27/36—Glass electrodes
-
- 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
-
- 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
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Nanotechnology (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of immunoassay methods based on the Prussian blue self energizing electrochemical sensor building of aluminium-, it is first to prepare Prussian blue modified electrode, again by itself and aluminium electrode, digital multimeter, reaction tank series connection is built into the self energizing electrochemical sensor, glucose oxidase and antibody mutual load are formed into enzyme mark nano-probe on nanogold particle again, it is formed into immuno-sandwich structure with sample to be tested on ELISA Plate again, glucose catalysis is set to generate a large amount of hydrogen peroxide using the immuno-sandwich structure, hydrogen peroxide is injected in self energizing electrochemical sensor again, the current signal of Prussian blue color and sensor can be significantly changed to Prussian blue oxidation by it, to by reading the current value generated, or Prussian blue color change is observed to measure the concentration of object.The method of the present invention equipment is simple, easy to carry, and can be used for the detection of Multiple Antibodies, provides a kind of simple, sensitive and stable method for portable inspectiont.
Description
Technical field
The invention belongs to biochemical analysis technical fields, and in particular to one kind is based on the Prussian blue self energizing electrification of aluminium-
Learn the immunoassay method of sensor building.
Background technique
With the development of society, environmental monitoring, food safety and medical diagnosis on disease are increasingly valued by people, therefore send out
Quick, easy, the sensitive detection method of exhibition is still one of hot spot studied now.In modern analysis detection architecture, big portion
The standard detecting method divided is based on foundation such as optics, chromatography, mass spectrums, these detection methods have accurate, sensitive, stable etc.
Advantage.But expensive equipment, complicated operation etc. limit its quick detection and in terms of further answer
With especially in remote districts.Therefore, design a kind of equipment it is simple, it is easily operated, stablize practical detection method have it is important
Meaning.
For self energizing electrochemical sensor as a kind of portable sensor, the electrode mainly modified by two forms one
A primary battery structure.The sensor itself can generate energy output, be not necessarily to other external energy, and the output of its energy and inspection
The concentration for surveying object in liquid has tight association.After this sensor is suggested, many researchs have been carried out.By in electricity
The different material of extremely upper modification, is successfully realized the detection to substances such as glucose, lactic acid, cholesterol.But it is traditional certainly at these
It energizes in electrochemical sensor, detection signal is often output power, and detection device is often also the electrochemical operation of profession
It stands, is unfavorable for its universal and portability.On the other hand, most of self energizing electrochemical sensor is typically split into cathode and anode
Two parts are detected, this is also unfavorable for its further integration.Therefore, simpler, small-sized self energizing is designed
Electrochemical sensor, and it is applied to quick, portable immunoassay field with important research meaning.
Summary of the invention
Based on background above, the purpose of the present invention is to provide one kind to be based on the Prussian blue self energizing electrochemical sensing of aluminium-
The immunoassay method of device building.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of immunoassay method based on aluminium-Prussian blue self energizing electrochemical sensor building, specifically include as
Lower step:
(1) preparation of Prussian blue modified electrode: by clean FTO glass electrode, platinum electrode and saturated calomel electrode
Respectively as working electrode, to electrode and reference electrode, be placed in containing 0.01-0.5 mol/L HCl, 0.01-0.5 mol/L
KCl、0.1-10 mmol/L K3[Fe(CN6)]、0.1-10 mmol/L FeCl3Electrolyte in, 0.1-0.6 V perseverance electricity
Pressure electropolymerization 300-400 seconds, is then washed with water, dries, and Prussian blue modified electrode is made；
(2) building of the Prussian blue self energizing electrochemical sensor of aluminium-: by the electricity of Prussian blue modification made from step (1)
Pole, aluminium electrode, digital multimeter series connection and a two interelectrode small-sized detection cell series connection, are made the Prussian blue confession of the aluminium-
It can electrochemical sensor；
(3) preparation of the nano-probe of glucose oxidase and antibody mutual load: by glucose oxidase and antibody according to
It is supported on nanogold particle by electrostatic adsorption, the enzyme mark nano-probe is made；
(4) formation of antibody-antigen-antibody interlayer type immune complex: in 96 hole polystyrene micropores of high-affinity
30-80 μ L antibody is added in plate, closes overnight, sequentially adds 30-80 μ L object, 30-80 μ L enzyme mark nano-probe shape
At antibody-antigen-antibody interlayer type immune complex；
(5) it is detected using the Prussian blue self energizing electrochemical sensor of aluminium-: gained interlayer type immune complex is used
After phosphate buffer solution washing, phosphate buffer (pH=6) of the 50-150 μ L containing 50 mmol/L glucose is added and is reacted,
Then reaction solution is injected into small-sized detection cell, the current signal in digital multimeter is read after 25 s, or directly observe
The color change of Prussian blue electrode measures the concentration of object.
The size of FTO glass electrode described in step (1) is 1 × 5 cm2；The aluminium electrode is to be pasted onto FTO by aluminium foil
Foil electrode is formed in glass electrode；
Prussian blue film is the circle of 6 mm of diameter on gained Prussian blue modified electrode, and blue, UV, visible light is presented
Optical absorption peak is at about 700 nm.
Small-sized detection cell described in step (2) is prepared by dimethyl silicone polymer (PDMS), and cross section is 6 mm's of diameter
Circle, volume are 40 μ L.
The partial size of nanogold particle described in step (3) is 16 nm.
The present invention is immunoreacted with the single chamber primary battery of the Prussian blue bipolar electrode building of aluminium-in conjunction with antigen-antibody, and building is certainly
Energize electrochemical immunosensor.Its technical principle is to be catalyzed glucose by introducing glucose oxidase on immunological probe
It decomposes and generates hydrogen peroxide, and hydrogen peroxide is injected in detection cell.It is Prussian blue to be reduced in the presence of there is no object
Colourless Prussia is white, and the Prussian blue primary battery of aluminium-releases lower current signal；In the presence of object, due to peroxidating
The presence of hydrogen, Prussia is white can be Prussian blue by the reoxidized generation of hydrogen peroxide, and increases the current signal of detection significantly
By force, wherein the variation of current signal and Prussian blue color are positively correlated with target concentration in a certain range, therefore can
With by digital multimeter come quantitative detection, can also be according to Prussian blue color change come qualitative detection.
Advantages of the present invention is as follows:
(1) material feedstock required for the present invention is easy to get, and preparation process is simple, low in cost.
(2) it is compared with conventional detection devices, digital multimeter can be used to quantify as signal reader for the present invention
Detection carries out qualitative detection with Prussian blue color change, accesses without extra power, greatlies simplify detection device,
It is easy to operate, it is suitable for portable inspectiont.
(3) present invention makes signal enhancing by introducing glucose oxidase catalysis breakdown of glucose generation hydrogen peroxide,
It can be used for the detection of Multiple Antibodies object, it is versatile, it is with a wide range of applications.
(4) detection method has the characteristics that high sensitivity, stability are strong, selectivity is good, the range of linearity is wide, and can
Detection for Multiple Antibodies.
Detailed description of the invention
Fig. 1 is that the present invention is based on the signals of the immunoassay method of the Prussian blue self energizing electrochemical sensor building of aluminium-
Figure；
Fig. 2 is the component pictorial diagram for constructing self energizing electrochemical sensor of the present invention；
Fig. 3 is by the standard working curve that does in embodiment 1 to alpha-fetoprotein；
Fig. 4 is the corresponding Prussian blue electrode color of standard sample of various concentration in example example 1.
Specific embodiment
In order to make content of the present invention easily facilitate understanding, With reference to embodiment to of the present invention
Technical solution is described further, but the present invention is not limited only to this.
Embodiment 1
1. the preparation of Prussian blue modified electrode
Using clean FTO glass electrode, platinum electrode and saturated calomel electrode as working electrode, to electrode and ginseng
Than electrode, it is placed in containing 0.1 mol/L HCl, 0.1 mol/L KCl, 5 mmol/L K3[Fe(CN6)]、5 mmol/L FeCl3
Electrolyte in, then electropolymerization 360 seconds under the constant voltage of 0.4 V are washed with water, dry, Prussian blue modification electricity is made
Pole.
2. self energizing electrochemical sensor constructs
By the Prussian blue modified electrode prepared, aluminium electrode, digital multimeter series connection and a two interelectrode small-sized inspections
Pond series connection is surveyed, the Prussian blue self energizing electrochemical sensor of the aluminium-is made.
3. the preparation of enzyme labelled antibody nano-probe
Firstly, 5 mL nano-Au solutions sodium carbonate is adjusted pH to 8.5, the first tire of 50 μ L, 0.5 mg/mL are added
The glucose oxidase of protein antibodies and 200 μ L, 0.5 mg/mL, 4 DEG C of incubated overnights.Then by above-mentioned solution 4 DEG C,
It is centrifuged 20 minutes under the conditions of 14000 rmp, the precipitating that centrifugation obtains is dispersed in 2 mL again and contains 1.0 wt % BSA and 0.1
In the PBS (pH=7.4) of wt % sodium azide, be stored in 4 DEG C it is spare.
4. the preparation of interlayer type immune complex
Firstly, the alpha-fetoprotein antibody of 50 μ L, 10 μ g/mL are added in the high affine plate in 96 holes, hatching overnight at 4 DEG C,
Add 300 μ L, the BSA of 1.0 wt % is closed 1 hour.After being washed with PBS solution, it is sequentially added into 50 μ L alpha-fetoproteins
The enzyme mark nano-probe of the series of concentrations standard sample of specific antigen and 50 μ L reacts 30 minutes respectively, removes supernatant.
5. the detection pair alpha-fetoprotein specific antigen
The PBS buffer solution (pH=6) that 100 μ L contain 50 mM glucose is added in gained interlayer type immune complex, instead
It answers 30 minutes.Then, it in the detection cell for taking 40 μ L reaction solutions injection sensor, reacts 25 seconds, reads the electricity in digital multimeter
The variation of flow valuve or the Prussian blue electrode color of observation.Standard working curve of its gained electric current to alpha-fetoprotein specific antigen
As shown in figure 3, corresponding Prussian blue electrode color change is as shown in Figure 4.
As seen from Figure 3, the range of linearity is 5-200 ng/mL, and detection is limited to 1.67 ng/mL.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (3)
1. a kind of immunoassay method based on the Prussian blue self energizing electrochemical sensor building of aluminium-, it is characterised in that: described
Method includes the following steps:
(1) preparation of Prussian blue modified electrode: clean FTO glass electrode, platinum electrode and saturated calomel electrode are distinguished
As working electrode, to electrode and reference electrode, be placed in containing 0.01-0.5 mol/L HCl, 0.01-0.5 mol/L KCl,
0.1-10 mmol/L K3[Fe(CN6)]、0.1-10 mmol/L FeCl3Electrolyte in, under the constant voltage of 0.1-0.6 V
It electropolymerization 300-400 seconds, is then washed with water, dries, Prussian blue modified electrode is made；
(2) building of the Prussian blue self energizing electrochemical sensor of aluminium-: by Prussian blue modified electrode, aluminium made from step (1)
Electrode, digital multimeter series connection and a two interelectrode small-sized detection cell series connection, are made the Prussian blue self energizing electrification of the aluminium-
Learn sensor；
(3) preparation of the nano-probe of glucose oxidase and antibody mutual load: by glucose oxidase and antibody by quiet
Enzyme mark nano-probe is made on nanogold particle in Electro Sorb used load；
(4) formation of antibody-antigen-antibody interlayer type immune complex: in 96 porous polystyrene microporous plates of high-affinity
Be added 30-80 μ L antibody, overnight close, sequentially add 30-80 μ L object, 30-80 μ L enzyme mark nano-probe formed it is anti-
Body-Ag-Ab interlayer type immune complex；
(5) it is detected using the Prussian blue self energizing electrochemical sensor of aluminium-: gained antibody-antigen-antibody interlayer type is exempted from
After epidemic disease compound is washed with phosphate buffer solution, the phosphoric acid buffer containing 50 mmol/L glucose of 50-150 μ L, pH=6 is added
Liquid is reacted, and then reaction solution is injected into the Prussian blue self energizing electrochemical sensor of aluminium-, and number ten thousand is read after 25 s
With the current signal on table, or observe the color change of Prussian blue electrode directly to measure the concentration of object.
2. the immunoassay side according to claim 1 based on the Prussian blue self energizing electrochemical sensor building of aluminium-
Method, it is characterised in that: the circle that the Prussian blue film of gained is 6 mm of diameter in step (1).
3. the immunoassay side according to claim 1 based on the Prussian blue self energizing electrochemical sensor building of aluminium-
Method, it is characterised in that: the partial size of nanogold particle described in step (3) is 16 nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810438328.XA CN108627559B (en) | 2018-05-09 | 2018-05-09 | Immunoassay method based on the Prussian blue self energizing electrochemical sensor building of aluminium- |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810438328.XA CN108627559B (en) | 2018-05-09 | 2018-05-09 | Immunoassay method based on the Prussian blue self energizing electrochemical sensor building of aluminium- |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108627559A CN108627559A (en) | 2018-10-09 |
| CN108627559B true CN108627559B (en) | 2019-08-09 |
Family
ID=63692369
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810438328.XA Expired - Fee Related CN108627559B (en) | 2018-05-09 | 2018-05-09 | Immunoassay method based on the Prussian blue self energizing electrochemical sensor building of aluminium- |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108627559B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109239135B (en) * | 2018-10-24 | 2021-04-27 | 福州大学 | Portable biological immunoassay method constructed based on flexible air pressure sensor |
| CA3120917A1 (en) * | 2018-11-27 | 2020-06-04 | Module Innovations Private Limited | Method for testing of antibiotic susceptibility in microorganisms |
| CN110068570B (en) * | 2019-04-03 | 2021-05-25 | 济南大学 | Preparation of electrochemical sensor with instant visual colorimetric display |
| CN110261453A (en) * | 2019-05-20 | 2019-09-20 | 武汉中科志康生物科技有限公司 | A kind of avian influenza virus electrochemical sensor based on screen printing electrode |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102053161A (en) * | 2009-10-29 | 2011-05-11 | 南京大学 | Disposable multi-channel electrochemical immunosensor with high sensitivity |
| CN102411022A (en) * | 2011-08-03 | 2012-04-11 | 中国科学院化学研究所 | Electrochemical sensor based on primary battery and preparation method thereof |
| CN106104264A (en) * | 2014-01-21 | 2016-11-09 | 加利福尼亚大学董事会 | Saliva biosensor and biological fuel cell |
-
2018
- 2018-05-09 CN CN201810438328.XA patent/CN108627559B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102053161A (en) * | 2009-10-29 | 2011-05-11 | 南京大学 | Disposable multi-channel electrochemical immunosensor with high sensitivity |
| CN102411022A (en) * | 2011-08-03 | 2012-04-11 | 中国科学院化学研究所 | Electrochemical sensor based on primary battery and preparation method thereof |
| CN106104264A (en) * | 2014-01-21 | 2016-11-09 | 加利福尼亚大学董事会 | Saliva biosensor and biological fuel cell |
Non-Patent Citations (2)
| Title |
|---|
| A bi-functional device for self-powered electrochromic window and self-rechargeable transparent battery applications;Jinmin Wang et al.;《NATURE COMMUNICATIONS》;20140923;第5卷 * |
| Self-powering amperometric sensor and biosensor;Lai Peng Wong et al.;《Journal of Electroanalytical Chemistry》;20120306;第671卷;80-84 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108627559A (en) | 2018-10-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108627559B (en) | Immunoassay method based on the Prussian blue self energizing electrochemical sensor building of aluminium- | |
| Ji et al. | Smartphone-based integrated voltammetry system for simultaneous detection of ascorbic acid, dopamine, and uric acid with graphene and gold nanoparticles modified screen-printed electrodes | |
| Ji et al. | Smartphone-based cyclic voltammetry system with graphene modified screen printed electrodes for glucose detection | |
| Yu et al. | A new enzyme immunoassay for alpha-fetoprotein in a separate setup coupling an aluminium/Prussian blue-based self-powered electrochromic display with a digital multimeter readout | |
| Tang et al. | Sensitive enzymatic glucose detection by TiO 2 nanowire photoelectrochemical biosensors | |
| Sun et al. | Nanoelectrochemistry of mammalian cells | |
| WO2021087945A1 (en) | Method for preparing bifunctional hybrid film for tumor exosome self-calibration detection | |
| Shi et al. | Self-powered dual-mode sensing strategy based on graphdiyne and DNA nanoring for sensitive detection of tumor biomarker | |
| Rawal et al. | A comprehensive review of bilirubin determination methods with special emphasis on biosensors | |
| CN109187708A (en) | A kind of electrochemical method detecting protein acetyltransferase activity | |
| Chen et al. | A self-powered aptasensor using the capacitor-amplified signal of a photofuel cell and a portable digital multimeter readout | |
| CN110441295A (en) | One kind is based on ferritin encapsulation Ir (ppy)3Biosensor preparation method | |
| CN109254063A (en) | A kind of preparation method of the Prussian blue electrochemica biological sensor marker of support type | |
| CN105044179B (en) | A kind of three-dimensional grapheme modified electrode detecting tumor markers and preparation method thereof | |
| CN110018210A (en) | A method of directly quickly detecting uric acid | |
| Deng et al. | Lab on smartphone with interfaced electrochemical chips for on-site gender verification | |
| CN101532980B (en) | Enzyme immunosensor for detecting Shigella species and its preparation method and application | |
| Ji et al. | Smartphone-based electrochemical system for biosensors and biodetection | |
| Tian et al. | A separated type cathode photoelectrochemical aptasensor for thrombin detection based on novel organic polymer heterojunction photoelectric material | |
| Lai et al. | A dry chemistry-based electrochemiluminescence device for point-of-care testing of alanine transaminase | |
| Jia et al. | Visual analysis of Alzheimer disease biomarker via low-potential driven bipolar electrode | |
| CN110082403A (en) | Based on gold-palladium nano flower/graphene composite material histone acetyltransferase timing-current sensor and its application | |
| Gao et al. | A dual functional analytical device for self-powered point of care testing and electric energy storage | |
| CN108760861A (en) | A kind of Ni-MOF based electrochemical sensors for detecting uric acid | |
| Song et al. | A signal amplifying photoelectrochemical immunosensor based on the synergism of Au@ CoFe2O4 and CdS/NiCo2O4 for the sensitive detection of neuron-specific enolases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190809 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |