CN102654475A - Bioelectrochemical sensor used for detecting hydrogen peroxide and manufacturing method thereof - Google Patents
Bioelectrochemical sensor used for detecting hydrogen peroxide and manufacturing method thereof Download PDFInfo
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- CN102654475A CN102654475A CN2012100793362A CN201210079336A CN102654475A CN 102654475 A CN102654475 A CN 102654475A CN 2012100793362 A CN2012100793362 A CN 2012100793362A CN 201210079336 A CN201210079336 A CN 201210079336A CN 102654475 A CN102654475 A CN 102654475A
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Abstract
The invention relates to a bioelectrochemical sensor used for detecting hydrogen peroxide and a manufacturing method of the bioelectrochemical sensor. The novel bioelectrochemical sensor is a sensor with a three-electrode system, wherein the counter electrode is a platinum electrode, the reference electrode is saturated calomel electrode and the working electrode is gold electrode. The sensor is based on orientation combination of a polypeptide horse radish peroxidase with characteristic sequence, the promotion effect of the polypeptide horse radish peroxidase on the activity of the enzyme and the characteristic of high sensitivity of electrochemical detection methods. The sensitivity of hydrogen peroxide sensors is effectively improved and satisfactory results are obtained.
Description
Technical field
The present invention relates to a kind of new bio electrochemical sensor and preparation method thereof, particularly a kind of new bio electrochemical sensor that detects hydrogen peroxide and preparation method thereof.
Technical background
Hydrogen peroxide (H
2O
2) all play an important role in many fields such as food, pharmacy, industry, biology, clinical practice and environmental engineerings, like food processing, paper-making industry, sterilization etc.Yet hydrogen peroxide is one of acid rain formative factor as a kind of atmosphere pollution.In addition, hydrogen peroxide is the accessory substance of cell ageing, death and some pathological state cellular metabolisms.Therefore, detecting hydrogen peroxide fast, delicately is the problem that scientific circles pay close attention to for a long time.At present; The method that detects hydrogen peroxide mainly contains titrimetry, AAS, chemoluminescence method, fluorometry and electrochemical method; Wherein electrochemical method is compared other analyzing detecting methods, has cheap device, advantage such as highly sensitive, simple and efficient.
The electrochemica biological sensor of developing based on various electrochemical method be one type with electrode as signal converter, the biology sensor of measuring with current potential or electric current.Electrochemical system is realized inputing or outputing of electric energy by electrode, thereby obtains the electric signal of electrode face finish material, and commonly used is three-electrode system.Three-electrode system comprises working electrode, auxiliary electrode (also claiming electrode) and contrast electrode, wherein be platinum electrode to electrode, contrast electrode is a saturated calomel electrode, working electrode is a gold electrode, flow through working electrode and to electrode of electric current.The measured current potential of working electrode is for contrast electrode.
In various types of electrochemica biological sensors, enzyme biologic sensor was because advantages such as the selectivity of enzyme self and sensitivity had obtained great development in recent years.The fixing means of enzyme has directly determined the height of enzyme activity in sensor, thereby affects the detection performance of enzyme biologic sensor significantly.As everyone knows, the catalytic center of most of enzymes is buried inner at zymoprotein, and owing to zymoprotein in reasons such as the irregular orientation of electrode surface and absorption sex change, the big molecule of zymoprotein is showing slower out-phase electron transport speed without the electrode surface of modifying.For solving this difficult problem; The researchist has been developed the modified electrode of a lot of types; Utilize various membrane materials to fix zymoprotein, and quicken the electron transport between enzyme and the electrode for zymoprotein provides a suitable microenvironment such as surfactant, polymkeric substance, lipid.Yet the distribution of enzyme in these decorative materials remains more unordered.Therefore, enzyme being fixed on the electrode oriented and orderedly, thereby realizing the optimization of enzymatic activity, is development novel enzyme biology sensor problem demanding prompt solution.
Summary of the invention
One of the object of the invention is to provide a kind of bioelectrochemical sensor that detects hydrogen peroxide; This sensor combines the specificity of horseradish peroxidase (HRP) based on the polypeptide matrix and to the facilitation of enzymatic activity; Realize that enzyme is directed in the specificity of electrode surface in the enzyme sensor, thereby reach the purpose that improves sensor sensitivity.
Two of the object of the invention is to provide the preparation method of this sensor.
For achieving the above object, the present invention adopts following mechanism: the polypeptide of some particular sequence can with some enzymes, combine like the specific site specificity of HRP, through the orientation of adjustment enzyme, make outside the activated centre of enzyme is exposed to, thereby promote the activity of enzyme.
According to above-mentioned mechanism, the present invention adopts following technical scheme:
A kind of bioelectrochemical sensor that detects hydrogen peroxide; Be the three-electrode system sensor; Be platinum electrode to electrode wherein, contrast electrode is a saturated calomel electrode, and working electrode is a gold electrode; It is characterized in that described gold electrode surfaces is built with the polypeptide matrix, horseradish peroxidase HRP is fixed on the electrode through polypeptide matrix orientation; The sequence of described polypeptide is: AHKVVPQRQIRHAYNRYGSC; The mol ratio of described polypeptide and horseradish peroxidase is 5:1～1:5.
A kind of method for preparing the bioelectrochemical sensor of above-mentioned detection hydrogen peroxide is characterized in that the concrete steps of this method are:
A. the gold electrode that will handle places 0.5 M H
2SO
4In, in 0-1.6 V voltage range, carry out cyclic voltammetry scan, sweep speed and be set to 100 mV/s, until reaching stable, at the nitrogen atmosphere inner drying;
B. step a gained gold electrode is immersed in the polypeptide solution, room temperature was placed 15～20 hours; Ultrapure water flushing then; To remove the not covalently bound polypeptide of gold surface; Subsequently, with electrode be immersed in contain 1 mM sulfydryl hexanol, pH is in N-(2-hydroxyethyl) piperazine-N'-2-ethane sulfonic acid HEPES solution of 6.0 0.5～1 hour, wash with ultrapure water; At the nitrogen atmosphere inner drying, promptly obtain peptide modified gold electrode; Described polypeptide solution contains 1 μ M-10 μ M polypeptide, 20 mM HEPES and 10 mM TCEP, and its pH is 6.0;
C. under air-tight state, the resulting gold electrode of step b is immersed in the HRP solution of 1 mg/mL-10 mg/mL incubated at room 2 .5～3.5h; With the ultrapure water washing, promptly obtain required working electrode again;
D. the resulting working electrode of step c is formed the bioelectrochemical sensor of three-electrode system with platinum electrode and saturated calomel electrode.
The disposal route of above-mentioned gold electrode is specially: pending gold electrode is placed Piranha solution, the i.e. concentrated sulphuric acid: 30%H
2O
2Volume ratio be 3: 1, middle immersion 2 minutes is adsorbed on the organism of electrode surface with removal, fully rinses well with ultrapure water; Subsequently gold electrode is polished with the thin emery paper of 5000 purposes; Be respectively 1 μ m containing granularity respectively then; 0.3 μ m is polished to minute surface successively on the silk of the alumina mortar of 0.05 μ m, the ultrasonic cleaning 5 minutes in the second alcohol and water respectively of the gold electrode after the polishing.
A kind of method that detects hydrogen peroxide; Adopt above-mentioned bioelectrochemical sensor to detect; The concrete steps that it is characterized in that this method are: under inert atmosphere, the working electrode immersion is contained in the damping fluid of liquid to be detected, adopt cyclic voltammetry or timing ampere method to carry out galvanochemistry scanning under the room temperature; It is 7.0 PBS damping fluid that described damping fluid adopts 100 mM, pH, wherein adds the o-phenylenediamine that 10 mM are arranged; Sweep speed when adopting cyclic voltammetry is 100 mV/s, and scanning voltage is-0.4 V～-0.8 V; Operating potential-583 mV when adopting timing ampere method.
The present invention chooses to be had the polypeptide of facilitation to the enzymatic activity of HRP (peptide sequence is: AHKVVPQRQIRHAYNRYGSC); Make up a matrix of this polypeptide in gold electrode surfaces; Utilize the specificity combination of polypeptide and HRP then, the HRP orientation is fixed to electrode surface.In testing process, in system to be detected, introduced a kind of electronic conductor---o-phenylenediamine, it plays the function served as bridge of electron transport between enzyme and electrode.Under the effect of HRP, o-phenylenediamine provides the electron reduction hydrogen peroxide, self is oxidized to 2,2 '-two amido azobenzenes with electrochemical activity, can be detected by electrochemical method, thereby obtain satisfactory detection result.
The present invention has made up a kind of novel hydrogen peroxide bioelectrochemical sensor that utilizes polypeptide matrix and horseradish peroxidase.The polypeptide that it is based on some specific sequence combines the orientation of horseradish peroxidase and to the facilitation of its enzymatic activity; Enzyme is fixed on electrode surface oriented and orderedly; Thereby solved the directionality problem of the enzyme that traditional enzyme electrochemical sensor cann't be solved; Promoted the electron transport of enzyme, thereby greatly improved the sensitivity that hydrogen peroxide detects at electrode surface.The present invention is significant in fixing this problem of the orientation of electrode surface for enzyme.
Description of drawings
Fig. 1 is the cyclic voltammogram at 100 mM PBS (pH 7.0) solution that contains 10 mM o-phenylenediamines, and wherein a is for only having modified the gold electrode of polypeptide; B is for modified the gold electrode of HRP and polypeptide simultaneously.
Fig. 2 is in containing 100 mM PBS (pH 7.0) solution of 10 mM o-phenylenediamines, the cyclic voltammogram of HRP and peptide modified gold electrode, and wherein a is not for containing H
2O
2B is for containing 1 mM H
2O
2Under the condition.
Fig. 3 adds H continuously in 100 mM PBS (pH 7.0) solution that contains 10 mM o-phenylenediamines
2O
2(concentration 1 * 10
-9～2 * 10
-4M) under the condition, the timing ampere curve of HRP and peptide modified gold electrode.Insertion figure is H
2O
2Concentration is 1 * 10
-9～1 * 10
-5Timing ampere curve in the M scope.
Fig. 4 is H in the damping fluid
2O
2The linear relationship of concentration and electric current.
Embodiment
Embodiment one: the preparation of peptide modified gold electrode
Pending gold electrode is placed the Piranha solution (concentrated sulphuric acid: 30%H
2O
2=3: soak 1) and be adsorbed on the organism of electrode surface with removal in 2 minutes, fully rinse well with ultrapure water.Subsequently gold electrode is polished with thin emery paper (5000 order); (granularity is respectively 1 μ m containing aluminium oxide respectively then; 0.3 μ m, 0.05 μ m) is polished to minute surface successively on the silk of mortar, the ultrasonic cleaning 5 minutes in the second alcohol and water respectively of the gold electrode after the polishing.At last electrode is placed 0.5 M H
2SO
4In, in 0-1.6 V potential range, carry out cyclic voltammetry scan to stable.At the nitrogen atmosphere inner drying, obtain the naked gold electrode of cleaning surfaces, can be used for the modification of polypeptide matrix.The Eppendorf pipe back-off that will contain 70 μ L polypeptide solutions (pH 6.0 for 10 μ M polypeptide, 20 mM HEPES and 10 mM TCEP) is on gold electrode; Guarantee that decorating liquid covers gold surface fully; Room temperature was placed after 16 hours, with the ultrapure water flushing, to remove the not covalently bound polypeptide of gold surface.Subsequently, electrode immersed in the 20 mM HEPES solution (pH 6.0) contain 1 mM sulfydryl hexanol 0.5 hour,,, promptly obtain peptide modified gold electrode at the nitrogen atmosphere inner drying with the ultrapure water flushing.
Embodiment two: horseradish peroxidase (HRP) is in the modification of gold electrode surfaces
The HRP solution for preparing 10 μ L are dropped to the gold electrode surfaces of successful modified polypeptide, and incubated at room 3 h are covered with the Eppendorf pipe at electrode surface, to prevent the enzyme solutions volatilization.Modifying good electrode rinses out with the material of ultrapure water with the weak absorption in surface before use.
Embodiment three: the obtaining of working electrode electrochemical signals
Because the electric conductivity of polypeptide is relatively poor, for HRP and interelectrode electron transport certain iris action is arranged, so modified electrode can't obtain the Direct Electrochemistry signal of HRP in 100 mM PBS damping fluids (pH7.0).Therefore, we have introduced a kind of electronic conductor---o-phenylenediamine in system.It can play the function served as bridge of electron transport between enzyme and electrode.Can see by Fig. 1; The gold electrode of only having modified polypeptide has no electrochemical response in containing 100 mM PBS damping fluids (pH7.0) of 10 mM o-phenylenediamines, and a pair of redox peak stable, the peak shape symmetry has appearred in the gold electrode of having modified simultaneously after the HRP.At 100 mVs
-1Sweep speed down, its reduction peak and oxidation peak current potential are respectively-583 mV and-547 mV, with the electrochemical parameter basically identical of 2,2 '-two amido azobenzenes of reporting in the document.Its apparent standard electrode potential (E
0 ') be-565 mV, peak separation (Δ E) is 36 mV.
Embodiment four: the catalytic performance research of working electrode
As shown in Figure 2, the H of adding 1 mM in 100 mM PBS damping fluid (pH7.0) systems that contain 10 mM o-phenylenediamines
2O
2After, HRP and peptide modified gold electrode obtain a pair of very significantly redox peak, and the catalytic activity owing to HRP is described, and body series can be to the H that exists in the solution
2O
2Produce sensitive current-responsive.
Embodiment five: the detection of variable concentrations hydrogen peroxide
In 100 mM PBS (pH 7.0) solution that contains 10 mM o-phenylenediamines, add H continuously
2O
2(concentration 1 * 10
-9～2 * 10
-4M) under the condition, HRP and peptide modified gold electrode timing ampere curve of gained under the operating potential of-583 mV are as shown in Figure 3.Along with H
2O
2The increase of concentration, the current-responsive that obtains also increases.Response current reaches stable state in 3 s, show the swift electron transmission has taken place on the electrode.Shown in Figure 4 is that HRP and peptide modified gold electrode are to variable concentrations H
2O
2The typical curve of response.At H
2O
2Concentration range is from 5.0 * 10
8MolL
-1To 1.0 * 10
4MolL
-1Between when changing, electric current and H
2O
2Concentration is linear, and the equation of linear regression formula is y=0.0087x+0.0374, R
2=0.9979, detect and be limited to 1.7 * 10
8MolL
-1(signal to noise ratio (S/N ratio) is 3).Therefore, the present invention can be used for the detection by quantitative of hydrogen peroxide.
Apparent Michaelis constant (
) combines a tolerance of tightness degree as enzyme and substrate; The affinity size that expressed enzyme and substrate combine can be used as and weighs the index that is fixed on the enzymatic activity on the electrode.
value is more little, and expression enzyme-to-substrate affinity is big more.According to the Lineweaver-Burk equation
Wherein, I
SsFor adding the steady current that obtains behind the substrate, C is for adding the concentration of substrate, I
MaxBe when adding the maximum current that substrate obtains when saturated, try to achieve apparent Michaelis constant (
) be 0.366 mmolL
-1For the electrochemical sensor of some non-directional immobilized enzymes, this numerical value is at 0.63 mmolL
-1To 23 mmolL
-1Between.Can explain that through more apparent Michaelis constant value the HRP among the present invention is because the directed fixation of polypeptide array and to H
2O
2Higher affinity is arranged, and this makes the present invention have clear superiority than the Traditional electrochemical sensor.
Among the present invention, it is 1.7 * 10 that the bioelectrochemical sensor of detection hydrogen peroxide can successfully be discerned least concentration
8MolL
-1Hydrogen peroxide, linear detection range is 5.0 * 10
8MolL
-1To 1.0 * 10
4MolL
-1, can be applicable to the Sensitive Detection of hydrogen peroxide.
Sequence table
< 110>Shanghai University
< 120>bioelectrochemical sensor of detection hydrogen peroxide and preparation method thereof
<160> 1
<210> 1
<211> 20
<212> PRT
< 213>artificial sequence
<400> 1
AHKVV PQRQI RHAYN RYGSC
Claims (4)
1. bioelectrochemical sensor that detects silver ion; Be the three-electrode system sensor; Be platinum electrode to electrode wherein, contrast electrode is a saturated calomel electrode, and working electrode is a gold electrode; It is characterized in that being modified with template DNA on the described gold electrode, the sequence of template DNA is: 5 '-CCTA CGACTGGATGACGATCCCTACGACTGAAAAAAAAAAAA-C
6-SH-3 ', this template DNA modification density on gold electrode is: 1.2 * 10
12Individual molecule/square centimeter～1.2 * 10
13Individual molecule/square centimeter.
2. method for preparing the biology sensor of detection silver ion according to claim 1, the concrete steps that it is characterized in that preparing the working electrode of this sensor are:
A. the gold electrode that will handle places 0.5 M H
2SO
4In, in 0～1.55 V voltage range, carry out cyclic voltammetry scan, sweep speed and be set to 100 mV/s, until reaching stable;
B. after drying up step a gained gold electrode with nitrogen, this gold electrode is immersed in the buffer solution, is inverted after 15～18 hours; Be immersed in again in the 1 mM sulfydryl hexanol WS and reacted 0.5～1.5 hour; Wash with ultrapure water, and dry up, promptly obtain the gold electrode that the template DNA chain is modified with nitrogen; Containing concentration in the described buffer solution is the template DNA chain of 1 μ M, the Tris-HCl of 10 mM, the TCEP of 10 mM and the NaCl of 0.1 M, and the pH of solution is 7.4.
3. method according to claim 2; The concrete steps that it is characterized in that the disposal route of described gold electrode are: drip 20 μ L Piranha solution in pending gold electrode surfaces; Be the concentrated sulphuric acid: the volume ratio of hydrogen peroxide is 3:1; React 2 min, rinse well with ultrapure water, nitrogen dries up; Gold electrode is polished on 5000 order sand paper after 5 min, be polished to minute surface successively containing on the silk of mortar of aluminium oxide that granularity is respectively 1 μ m, 0.3 μ m, 0.05 μ m, ultrasonic successively 5 min in ethanol, ultrapure water remove impurity then.
4. method that detects silver ion adopts the biology sensor of detection silver ion according to claim 1, it is characterized in that the concrete steps of this method are:
A. the working electrode gold electrode in the biology sensor is immersed in reaction buffer solution and the mixed liquor of solution to be measured by the volume ratio formation of 9:1; Described reaction buffer solution contains 100 nM primed DNAs, 0.5 U/ μ L BsaBI endonuclease, 0.05 U/ μ L Bst nucleic acid polymerase, 50 μ M dNTPs, 1 * NE buffering 4; Describedly be: 50 mM KAc, the Tris-HAc of 20 mMpH 7.9,10 mM Mg (Ac)
2, 55～65 degree reactions 5～10 minutes,, dry up with nitrogen with the ultrapure water flushing, obtain enzyme and cut the gold electrode of handling; Described primed DNA sequence is: 5 '-CAGTCCTAGG-3 ';
B. under inert atmosphere, the gold electrode of step a gained is put into the Electrochemical Detection damping fluid, described Electrochemical Detection damping fluid is for containing 50 μ M [Ru (NH
3)
6]
3+10 mM Tris-HCl, the detection damping fluid of pH=7.4 adopts cyclic voltammetry or timing coulometry to carry out galvanochemistry scanning, the reaction buffering is for adding 50 μ M [Ru (NH
3)
6]
3+10 mM pH, 7.4 Tris-HCl; Sweep speed when adopting cyclic voltammetry is 50 mV/s, and sweep limit is-0.45 V～0.05 V; Recurrence interval when adopting the timing coulometry is 250 ms, and initial potential 0.2 V stops current potential-0.5 V.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604772A (en) * | 2013-11-12 | 2014-02-26 | 浙江省计量科学研究院 | Method and device for measuring gas phase H2O2 concentration by using tunable laser absorption spectrum |
| CN108241013A (en) * | 2017-08-23 | 2018-07-03 | 河南大学 | The method that electrochemical sensing electrode based on polyaniline deposition quantitatively detects -1 activity of polyadenosine diphosphate ribose polymerase |
| CN109884154A (en) * | 2019-03-07 | 2019-06-14 | 贵州大学 | One kind being based on Fe2+-H2O2The electrochemical sensing assays method of o-phenylenediamine system |
| CN113008957A (en) * | 2021-01-20 | 2021-06-22 | 新乡医学院 | Method for manufacturing double microelectrodes capable of detecting hydrogen peroxide and nitric oxide in vivo synchronously |
| CN113514518A (en) * | 2021-03-30 | 2021-10-19 | 江苏大学 | Preparation method of hydrogen peroxide sensor based on polypeptide-enzyme composite material |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070231794A1 (en) * | 2005-09-21 | 2007-10-04 | Combimatrix Corporation | Process to detect binding events on an electrode microarray using enzymes |
| CN102128865A (en) * | 2010-11-28 | 2011-07-20 | 上海大学 | Method for detecting phenol compounds by using horseradish peroxidase-modified electrode |
| CN102608180A (en) * | 2012-03-21 | 2012-07-25 | 上海大学 | Bioelectrochemical sensor for detecting silver ion and preparation method thereof |
-
2012
- 2012-03-23 CN CN201210079336.2A patent/CN102654475B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070231794A1 (en) * | 2005-09-21 | 2007-10-04 | Combimatrix Corporation | Process to detect binding events on an electrode microarray using enzymes |
| CN102128865A (en) * | 2010-11-28 | 2011-07-20 | 上海大学 | Method for detecting phenol compounds by using horseradish peroxidase-modified electrode |
| CN102608180A (en) * | 2012-03-21 | 2012-07-25 | 上海大学 | Bioelectrochemical sensor for detecting silver ion and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| JING ZHAO,ET AL.: "An amperometric biosensor for the detection of hydrogen peroxide released from human breast cancer cells", 《BIOSENSORS AND BIOELECTRONICS》, vol. 41, 12 October 2012 (2012-10-12) * |
| JINGLIN FU,ET AL.: "Peptide-Modified Surfaces for Enzyme Immobilization", 《PLOS ONE》, vol. 6, no. 4, 8 April 2011 (2011-04-08) * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604772A (en) * | 2013-11-12 | 2014-02-26 | 浙江省计量科学研究院 | Method and device for measuring gas phase H2O2 concentration by using tunable laser absorption spectrum |
| CN108241013A (en) * | 2017-08-23 | 2018-07-03 | 河南大学 | The method that electrochemical sensing electrode based on polyaniline deposition quantitatively detects -1 activity of polyadenosine diphosphate ribose polymerase |
| CN109884154A (en) * | 2019-03-07 | 2019-06-14 | 贵州大学 | One kind being based on Fe2+-H2O2The electrochemical sensing assays method of o-phenylenediamine system |
| CN113008957A (en) * | 2021-01-20 | 2021-06-22 | 新乡医学院 | Method for manufacturing double microelectrodes capable of detecting hydrogen peroxide and nitric oxide in vivo synchronously |
| CN113514518A (en) * | 2021-03-30 | 2021-10-19 | 江苏大学 | Preparation method of hydrogen peroxide sensor based on polypeptide-enzyme composite material |
| CN113514518B (en) * | 2021-03-30 | 2023-05-09 | 江苏大学 | Preparation method of hydrogen peroxide sensor based on polypeptide-enzyme composite material |
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