CN107843631B - Protease detection electrochemical sensor and preparation method and detection method - Google Patents
Protease detection electrochemical sensor and preparation method and detection method Download PDFInfo
- Publication number
- CN107843631B CN107843631B CN201711421008.5A CN201711421008A CN107843631B CN 107843631 B CN107843631 B CN 107843631B CN 201711421008 A CN201711421008 A CN 201711421008A CN 107843631 B CN107843631 B CN 107843631B
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- protease
- detection
- electrochemical sensor
- preparation
- 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
- G01N27/327—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
-
- 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
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Hematology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Protease detection electrochemical sensor, the electrochemical sensor, which is used, modifies polypeptide-graphene complex of one layer of pyrene fourth label in glassy carbon electrode surface, the sequence signature of the polypeptide is: the polypeptide can be cut by corresponding protease, the third amino acid in cleavage site toward carbon teminal direction is histidine, and the 4th amino acid is the lysine that pyrene butyl group has been modified on side chain.The detection method of protease detection, comprising the following steps: 1: the preparation of electrochemical sensor, including following sub-step: 1.1: polypeptide-graphene complex preparation: 1.2: the preparation of electrochemical sensor: 2: the detection of protease.This method belongs to the high-throughput electrochemical sensing method of " signal enhancing ", has many advantages, such as that background current is low, strong antijamming capability.
Description
Technical field
The present invention relates to a kind of electrochemical sensor for protease detection and preparation method and detection method, belonging to
Learn detection technique field.
Background technique
Protease being capable of aminosal or peptide bonds in polypeptides.It is in the side such as digestion and absorption, Wound healing, function be immune
Face has decisive role.The generation of some major diseases and the content of protease and activity change are closely related, for example, cancer
Disease, Apoptosis, Alzheimer disease, AIDS etc..Therefore, diagnosing and treating of the protease detection in these major diseases
Research etc. is of great significance.Method currently used for detecting protease mainly has high performance liquid chromatography, gel chromatography, matter
Spectrum and fluorescence spectrum etc., but high performance liquid chromatography, gel chromatography, mass spectrum need that special instrument, sensitivity be lower, analysis cost
It is higher.Fluorescent spectrometry generally requires that enzyme substrate (usually polypeptide) both ends are marked, takes time and effort, in addition, polypeptide two
The efficiency of protease catalysis cutting is likely to affect after end is labeled.Between Protein cleavage reaction in research vital movement and disease
Disease prevention and treatment in significant role, establish it is a kind of it is inexpensive, simple and quick, suitable for monitoring protease in common laboratory or clinic
The method of content or activity change is one of hot spot of current research.
Electrochemical sensing technology is to convert change in electric for the concentration variation of test substance according to electrochemical principle
A kind of sensing technology.The outstanding feature of the technology is research information on the surface“It focuses”With the requirement pole of enhancing, sample
It is micro-, to isolate and purify simplicity, high sensitivity, selectivity good etc..The main of electrochemical sensor currently used for protease detection is set
Meter theory is: by the one of substrate polypeptide it is terminal modified on can generate group (such as ferrocene, methylene blue, the enzyme of electrochemical signals
Marker etc.), for generating electric signal, another terminal modified special amino acid (usually cysteine) is used for polypeptide
Fixed to electrode surface (usually gold electrode).After polypeptide is cut by protease, electric signal group will be from electrode surface
It falls off, leads to current reduction.But existing main problem is this kind of electrochemical sensing method in practical applications: first is that electricity
Signal group itself is easy degradation or rotten, and the long-time for being unfavorable for electrode chip saves；Second is that the inspection of this " signal decaying "
Survey mode has biggish background current, and sensitivity is lower, error is larger.Therefore, a kind of simple and sensitive, " signal enhancing " are developed
High-throughput electrochemical sensing method, be of great significance for the detection and the monitoring of some major diseases of protease.
Summary of the invention
It is an object of the invention to overcome the above problem present in current protease detection, a kind of protease inspection is provided
It surveys with electrochemical sensor and preparation method and detection method.
To achieve the purpose of the present invention, using following technical solutions: protease detection electrochemical sensor, it is described
Electrochemical sensor use glassy carbon electrode surface modify one layer of pyrene fourth label polypeptide-graphene complex, described is more
The sequence signature of peptide is: the polypeptide can be cut by corresponding protease, third ammonia of the cleavage site toward carbon teminal direction
Base acid is histidine, and the 4th amino acid is the lysine that pyrene butyl group has been modified on side chain.
The protease detection preparation method of electrochemical sensor, comprising the following steps:
A: polypeptide-graphene complex preparation: by the polypeptide solution ultrasonic mixing of graphene nanometer sheet and pyrene fourth label
30 minutes, the sequence signature of the polypeptide was: the polypeptide can be cut by corresponding protease, and cleavage site is toward carbon
The third amino acid of extreme direction is histidine, and the 4th amino acid is the lysine that pyrene butyl group has been modified on side chain, centrifugation
The remaining polypeptide in reject upper layer, then with phosphate buffer solution by obtained polypeptide-graphene complex centrifuge washing 3 times,
It is saved backup at 4 DEG C；
B: the preparation of electrochemical sensor: polypeptide-graphene complex that the pyrene fourth being prepared in step A is marked is more
It is shaken and is dispersed with phosphate buffer solution, taken out 5 microlitres of mixed liquors and be added drop-wise to the glassy carbon electrode surface that diameter is 3 mm, dry in the air naturally
It is dry, obtain electrochemical sensor.
The detection method of protease detection, comprising the following steps:
1: the preparation of electrochemical sensor, including following sub-step:
1.1: the preparation of polypeptide-graphene complex: the polypeptide solution ultrasound that graphene nanometer sheet is marked with pyrene fourth being mixed
It closes 30 minutes, the sequence signature of the polypeptide is: the polypeptide can be cut by corresponding protease, and cleavage site is past
The third amino acid in carbon teminal direction is histidine, and the 4th amino acid is the lysine that pyrene butyl group has been modified on side chain, from
The remaining polypeptide in heart reject upper layer, then with phosphate buffer solution by obtained polypeptide-graphene complex centrifuge washing 3
It is secondary, it is saved backup at 4 DEG C；
1.2: the preparation of electrochemical sensor: polypeptide-graphene complex that the pyrene fourth being prepared in step A is marked
Multi-purpose phosphate buffer solution concussion dispersion takes out 5 microlitres of mixed liquors and is added drop-wise to the glassy carbon electrode surface that diameter is 3 mm, dries in the air naturally
It is dry, obtain electrochemical sensor；
2: the detection of protease:
The electrode that step 1 is obtained is soaked in protein enzyme solution to be detected, is rinsed well after taking-up with secondary water, then
Cyclic voltammetry scan test, the phosphate buffer solution concentration are carried out in the phosphate buffer solution comprising 50 μM of copper sulphate
For 0.2 M, pH value 7.4；
Further, cyclic voltammetry scan uses three-electrode system, and the electrochemical sensor that step 1 obtains is work electricity
Pole, the Ag/AgCl electrode of saturation are reference electrode, and Pt electrode is auxiliary electrode.
The advantages of this technology is invented and effect are as follows: (1) preparation process of sensing electrode is simple, cost is relatively low；(2) it is not required to
Telecommunications labelled notation is carried out to protease-based bottom in advance, electric signal directly can produce by the oxidation of electro-catalysis hydrone, have
Have that error is small, advantages of environment protection；(3) polypeptide fragment after digestion is cut can be complexed with copper ion, and formation can be with catalytic water
The elctro-catalyst of oxidation has high sensitivity, advantages of environment protection；(4) this method belongs to the high-flux electric of " signal enhancing "
Chemical sensitisation method, with the increase of protease concentration, electric signal is gradually increased, the low, strong antijamming capability with background current
The advantages that, the Electrochemical Detection for succeeding in developing achievable multiple types protease of the technology.
Detailed description of the invention
Fig. 1 is cyclic voltammogram of the sensor when impregnating by and without caspase-3 solution.
Fig. 2 is the linearity curve to caspase-3 detection.
Fig. 3 is the selectivity to caspase-3 detection.
Fig. 4 is the linearity curve to trypsase detection.
Specific embodiment
In order to more fully explain implementation of the invention, embodiment of the invention is provided.These embodiments are only
Elaboration to the technique, does not limit the scope of the invention, and is illustrated in the present invention with following embodiment, but be not limited to following implementations
Example, any change are included in technical scope of the invention.
The sequence signature of polypeptide used in the present invention is: the polypeptide can be cut by corresponding protease, and
The third amino acid in cleavage site toward carbon teminal direction is histidine, and the 4th amino acid is that pyrene butyl group has been modified on side chain
Lysine, corresponding protease, that is, protease to be detected.In Fig. 1, curve a is polypeptide (Ac-GDEVDSGHK-Pyrene fourth)-stone
The circulation volt curve that the electrode of black alkene compound modification obtains when impregnating not in caspase-3 solution, curve b is the modification
Electrode impregnates after a certain period of time in caspase-3 solution, then the circulation scanned in the phosphate buffer solution comprising copper ion
Volt-ampere curve, the concentration of caspase-3 are 50 ng/mL.Fig. 2 is the oxidation current value and caspase-3 concentration at 0.8 V
Linearity curve, the concentration of caspase-3 is successively 0.05,0.5,5,10,20 ng/mL.Fig. 3 is the selection of sensor
Property.It is corresponding in turn to from 1 to 7: blank control experiment, 20 ng/mL caspase-3,20 ng/mL bovine serum albumins, 20
Ng/mL fibrin ferment, 20 ng/mL trypsase, 20 ng/mL beta-secretases, 10% serum.Fig. 4 is polypeptide (Ac-GKGGHK-
Pyrene fourth)-graphene complex modified electrode linearity curve that trypsase is detected.Polypeptide (Ac-GDEVDSGHK-Pyrene fourth) and
Polypeptide A c-GKGGHK-Pyrene fourth is the production of Shanghai Qiang Yao biotechnology company.
Embodiment 1:
1: the preparation of electrochemical sensor:
1.1: the preparation of polypeptide-graphene complex: it includes 0.1 mM that 1 mL is added into 0.5 mg graphene nanometer sheet
Polypeptide (Ac-GDEVDSGHK-Pyrene fourth) phosphate buffer solution (10 mM, pH 7.4), ultrasonic mixing 30 minutes, be centrifuged reject
The remaining polypeptide in upper layer, then with phosphate buffer solution by obtained polypeptide-graphene complex centrifuge washing 3 times, in 4
It is saved backup at DEG C；
The preparation of 1.2 electrochemical sensors: the polypeptide being prepared in step 1.1-graphene complex phosphoric acid is delayed
Solution concussion dispersion is rushed, 5 microlitres of mixed liquors are taken out, the glassy carbon electrode surface that diameter is 3 mm is added drop-wise to, dries in the air naturally at room temperature
It is dry, obtain polypeptide (Ac-GDEVDSGHK-Pyrene fourth)-graphene complex modification electrode, i.e., electrochemical sensor of the invention；
The detection of 2:caspase-3
By polypeptide (Ac-GDEVDSGHK obtained in 1-Pyrene fourth)-graphene complex modification electrode respectively comprising not
It is impregnated 20 minutes in the phosphate buffer solution of concentration c aspase-3, gently rinses electrode surface with secondary water, then electrode is soaked
It steeps and carries out cyclic voltammetry scan in the phosphate buffer solution (0.2 M, pH 7.4) comprising 50 μM of copper sulphate, scanning speed is
50 mV/s, scanning range are 0.3 ~ 1.0V.It will be seen from figure 1 that polypeptide (Ac-GDEVDSGHK-Pyrene fourth)-graphene is compound
The electrode of object modification does not have redox peaks in electrolyte solution, and (curve a) shows that the modified electrode cannot be catalyzed hydrone
Oxidation when the electrode impregnates after a certain period of time in caspase-3 solution, it is (bent still to there is an apparent oxidation peak
Line b), after showing that the polypeptide of graphene surface is cut, the polypeptide fragment (SGHK- pyrene fourth) for remaining in electrode surface can be with copper
Ion forming complex, to be catalyzed the electroxidation of hydrone.Fig. 2 is the line between oxidation current value and caspase-3 concentration
Sexual intercourse, it can be seen from the figure that current value increases with the increase of caspase-3 concentration.Therefore, which can be with
For the detection of caspase-3, detection is limited to 0.01 ng/mL, and the range of linearity is 0.05 ~ 20 ng/mL.
Embodiment 2: to the selectivity of caspase-3 detection:
By embodiment: the caspase-3 in 1 changes substance to be tested into, and the condition of other steps does not change, is prepared
Each working electrode is tested.Experimental result is as shown in figure 3,1 to 7 are corresponding in turn to: blank control experiment, 20 ng/mL
Caspase-3,20 ng/mL bovine serum albumins, 20 ng/mL fibrin ferments, 20 ng/mL trypsase, 20 ng/mL beta-secretases
Enzyme, 10% serum.It can be seen from the figure that basic using the electric signal of 3 ~ 7 protein electric signal generated and blank control
Equally, illustrate the method property of can choose detection caspase-3 of the invention.
Embodiment 3: the detection of trypsase
By polypeptide (Ac-GDEVDSGHK in embodiment 1-Pyrene fourth) it is changed to polypeptide (Ac-GKGGHK-Pyrene fourth), obtain polypeptide
(Ac-GKGGHK-Pyrene fourth)-graphene modified electrode, it is then molten in the phosphoric acid buffer comprising various concentration trypsase respectively
It is impregnated 20 minutes in liquid.Other steps and experiment condition are consistent with the detection of caspase-3 in embodiment 1.Fig. 4 be current value with
The linear relationship of trypsinase concentration.It can be seen from the figure that current value increases with the increase of trypsinase concentration, linearly
Range is 1 ~ 500 ng/mL, and detection is limited to 0.5 ng/mL.Caspase-3 in use embodiment 3, bovine serum albumin,
Fibrin ferment, beta-secretase, when 10% serum does control experiment, the current value of resulting electric signal and blank control experiment is basic
Equally, illustrate that the modified electrode has selectivity well to the detection of trypsase.Therefore, if by the more of graphene surface
Peptide changes the polypeptide fragment specifically responded to a certain protease into, and this method can be used for the Electrochemical Detection of corresponding protease.
Sequence table
<110>Anyang Teachers College
<120>protease detection electrochemical sensor and preparation method and detection method
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 9
<212> PRT
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
Gly Asp Glu Val Asp Ser Gly His Lys
1 5
<210> 2
<211> 6
<212> PRT
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 2
Gly Lys Gly Gly His Lys
1 5
Claims (3)
1. the preparation method that electrochemical sensor is used in protease detection, it is characterised in that the following steps are included:
A: the preparation of polypeptide-graphene complex: the polypeptide solution ultrasonic mixing 30 that graphene nanometer sheet is marked with pyrene fourth is divided
The sequence signature of clock, the polypeptide is: the polypeptide can be cut by corresponding protease, and cleavage site is toward carbon teminal direction
Third amino acid be histidine, the 4th amino acid be modified on side chain pyrene butyl group lysine, be centrifuged reject on
The remaining polypeptide of layer, then with phosphate buffer solution by obtained polypeptide-graphene complex centrifuge washing 3 times, in 4 DEG C
Under save backup；
B: polypeptide-graphene complex of the pyrene fourth being prepared in step A label the preparation of electrochemical sensor: is mostly used phosphorus
The concussion dispersion of acid buffering solution takes out 5 microlitres of mixed liquors and is added drop-wise to the glassy carbon electrode surface that diameter is 3 mm, and naturally dry obtains
To electrochemical sensor.
2. the detection method of protease detection, it is characterised in that the following steps are included:
1: the preparation of electrochemical sensor, including following sub-step:
1.1: polypeptide-graphene complex preparation: by the polypeptide solution ultrasonic mixing 30 of graphene nanometer sheet and pyrene fourth label
Minute, the sequence signature of the polypeptide is: the polypeptide can be cut by corresponding protease, and cleavage site is toward carbon teminal side
To third amino acid be histidine, the 4th amino acid be modified on side chain pyrene butyl group lysine, be centrifuged reject
The remaining polypeptide in upper layer, then with phosphate buffer solution by obtained polypeptide-graphene complex centrifuge washing 3 times, in 4
It is saved backup at DEG C；
1.2: the preparation of electrochemical sensor: polypeptide-graphene complex that the pyrene fourth being prepared in step A marks being mostly used
Phosphate buffer solution concussion dispersion, take out 5 microlitres of mixed liquors be added drop-wise to diameter be 3 mm glassy carbon electrode surface, naturally dry,
Obtain electrochemical sensor；
2: the detection of protease:
The electrode that step 1 is obtained is soaked in protein enzyme solution to be detected, is rinsed well after taking-up with secondary water, then is being wrapped
Cyclic voltammetry scan test is carried out in phosphate buffer solution containing 50 μM of copper sulphate, the phosphate buffer solution concentration is 0.2
M, pH value 7.4.
3. the detection method of protease detection according to claim 2, it is characterised in that: cyclic voltammetry scan is using three electricity
Polar body system, the electrochemical sensor that step 1 obtains are working electrode, and the Ag/AgCl electrode of saturation is reference electrode, and Pt electrode is
Auxiliary electrode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711421008.5A CN107843631B (en) | 2017-12-25 | 2017-12-25 | Protease detection electrochemical sensor and preparation method and detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711421008.5A CN107843631B (en) | 2017-12-25 | 2017-12-25 | Protease detection electrochemical sensor and preparation method and detection method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107843631A CN107843631A (en) | 2018-03-27 |
| CN107843631B true CN107843631B (en) | 2019-08-23 |
Family
ID=61684421
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711421008.5A Expired - Fee Related CN107843631B (en) | 2017-12-25 | 2017-12-25 | Protease detection electrochemical sensor and preparation method and detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107843631B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109470757B (en) * | 2019-01-04 | 2020-04-03 | 安阳师范学院 | Electrochemical detection method for prostate specific antigen detection |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104049007A (en) * | 2014-04-15 | 2014-09-17 | 南昌大学 | Trypsin-chymotrypsin electrochemical synchronous detection method based on enzyme digestion |
| CN105274000A (en) * | 2014-07-15 | 2016-01-27 | 中国科学院大连化学物理研究所 | Immobilized enzyme reactor and preparation method and application |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103149185B (en) * | 2013-02-05 | 2014-12-10 | 苏州大学 | Novel high-efficiency protease activity detecting method |
| CN106520913B (en) * | 2016-09-22 | 2020-02-21 | 江苏大学 | Preparation method of graphene oxide-DNA sensor based on enzyme digestion cycle amplification and application of graphene oxide-DNA sensor in thrombin detection |
-
2017
- 2017-12-25 CN CN201711421008.5A patent/CN107843631B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104049007A (en) * | 2014-04-15 | 2014-09-17 | 南昌大学 | Trypsin-chymotrypsin electrochemical synchronous detection method based on enzyme digestion |
| CN105274000A (en) * | 2014-07-15 | 2016-01-27 | 中国科学院大连化学物理研究所 | Immobilized enzyme reactor and preparation method and application |
Non-Patent Citations (4)
| Title |
|---|
| A graphene oxide platform for energy transfer-based detection of protease activity;Juan Li;《Biosensors and Bioelectronics》;20110308;第26卷;全文 |
| A signal-on electrochemical strategy for protease detection based onthe formation of ATCUN-Cu(II);Ning Xia等;《Sensors and Actuators B: Chemical》;20160326;第232卷;第557页abstract，第558页Experimental |
| Electronic π‑Delocalization Boosts Catalytic Water Oxidation by Cu(II) Molecular Catalysts Heterogenized on Graphene Sheets;Pablo Garrido-Barros等;《Journal of the American Chemical Society》;20170830;第139卷;第12907-12910页 |
| Graphene Oxide–Peptide Conjugate as an Intracellular Protease Sensor for Caspase-3 Activation Imaging in Live Cells;Haibo Wang等;《Angewandte Chemie International Edition》;20110616;第50卷;第7065-7068页，图1 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107843631A (en) | 2018-03-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10605761B2 (en) | Electrochemical biosensor based on aptamer/nano silver probe and EXO I enzyme | |
| Pundir et al. | Determination of glycated hemoglobin with special emphasis on biosensing methods | |
| Li et al. | Development of a 3D origami multiplex electrochemical immunodevice using a nanoporous silver-paper electrode and metal ion functionalized nanoporous gold–chitosan | |
| CN101655473B (en) | Preparation method of nanogold immunoelectrode | |
| Wang et al. | A supersandwich multienzyme–DNA label based electrochemical immunosensor | |
| CN110441295A (en) | One kind is based on ferritin encapsulation Ir (ppy)3Biosensor preparation method | |
| CN104502583A (en) | Carbon nano tube/nanogold composite membrane electrochemical immunosensor and application thereof | |
| Zatloukalová et al. | Electrochemical Determination of Transmembrane Protein Na+/K+‐ATPase and Its Cytoplasmic Loop C45 | |
| CN104614417B (en) | A kind of electrochemical method for glycoprotein detection | |
| CN101532980B (en) | Enzyme immunosensor for detecting Shigella species and its preparation method and application | |
| CN107843631B (en) | Protease detection electrochemical sensor and preparation method and detection method | |
| CN108195913B (en) | A kind of biosensor and its construction method for Electrochemical Detection HER2 | |
| ANZAI et al. | Urea sensor based on an ion-sensitive field effect transistor. IV. Determination of urea in human blood | |
| CN104407138B (en) | A kind of preparation method of the biology sensor based on pine branch shape nm of gold copper and application | |
| EP4318694A2 (en) | Membraneless fuel cell | |
| CN105758912A (en) | Preparation and application of nano TiO2-MoS2 photoelectric Saos-2 cell sensor | |
| Deng et al. | Determination of selenomethionine in selenium-enriched yeast using capillary electrophoresis on-line coupled with electrochemiluminescence detection | |
| JP2638877B2 (en) | Immunochemical assay | |
| CN109470757B (en) | Electrochemical detection method for prostate specific antigen detection | |
| CN106053570B (en) | A kind of microcapsule algae toxin electrochemical detection method of graphene signal amplification | |
| WO2013039455A1 (en) | Amperometric sensor | |
| Zhang et al. | “Covalent biosensing” enables a one-step, reagent-less, low-cost and highly robust assay of SARS-CoV-2 | |
| Chen et al. | Ultrasensitive sensing urinary cystatin C via an interface-engineered graphene extended-gate field-effect transistor for non-invasive diagnosis of chronic kidney disease | |
| CN108007989B (en) | Detection method based on split polypeptide as recognition molecule | |
| Wang et al. | Amplified voltammetric strategy for the detection of proteases using dopamine-modified gold nanoparticles |
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: 20190823 Termination date: 20201225 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |