CN105784819B - A kind of fit electrode of interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide and preparation method and application - Google Patents
A kind of fit electrode of interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide and preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of fit electrode of interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide and preparation method and application.Nitrogen-doped graphene/molybendum disulfide complexes (3D G N/MoS2) negatively charged in the solution, horseradish peroxidase (HRP) is positively charged in the solution, and horseradish peroxidase/nitrogen-doped graphene/molybendum disulfide complexes (HRP/3D G N/MoS are combined into by electrostatic attraction active force2).Interferon is fit, and ssDNA passes through electrostatic attraction and π π intermolecular forces and HRP/3D G N/MoS2With reference to formation ssDNA/HRP/3D G N/MoS2The aptamer modified electrodes of/GCE.The linear detection range for electrode pair interferon that this is fit is 10‑14‑10‑9M, lowest detection are limited to 3.3 × 10‑15M.The measurement result of actual sample shows that this method has good applicability.
Description
Background technology
Gamma interferon (IFN-γ), it is that one kind passes through phase also referred to as " immune interferon " or " immunocyte interferon "
Close the cell factor of the transcriptional control activation multi-signal pathway of immunogene.Gamma interferon is by the T cell (chest that activates
Gland derivation cell) and NK (NK cells) secretion.Gamma interferon has immunoregulation, antiviral, antiproliferative and resisted
The effect of tumour.Compared with other cell factors, gamma interferon (2-5d) before disease incidence just changes significantly, therefore
Gamma interferon is a kind of important marker of early stage disease detection.
Concentration abnormality of the gamma interferon in human serum and urine is related to many diseases, such as cancer, AIDS and bone
Marrow leukaemia.Due to the important physiological property of interferon, develop that new, fast and effectively detection of the method to interferon has
Significance.Up to the present, many detection methods have been reported, as electrochemistry, SERS, colorimetric method,
Fluorescence method etc..Wherein, the method for electrochemistry aptamer sensor detection interferon causes larger research interest.Fit identification member
The combination selection adapted of part and various chemotrons, decides that selection of the electrochemistry aptamer sensor to target analyte detection is special
One property, sensitivity and accuracy of detection.The selection of electrode material, the sensitivity for raising sensor are most important.
Prior art has been disclosed for a variety of electrochemistry aptamer sensors and detection method for gamma interferon.Liu etc.
(Ying Liu, Nazgul Tuleouva, Erlan Ramanculov, and Alexander Revzin, Anal.Chem.,
2010,82,8131-8136) report on gold electrode, pass through the aptamer sensor of golden sulfide linkage formation determination gamma interferon, inspection
The detection for surveying gamma interferon is limited to 0.06nM.(Genping Yan, Yonghong Wang, the Xiaoxiao He, Kemin such as Yan
Wang, Jinquan Liu, Yudan Du, Biosensors and Bioelectronics, 2013,44,57-63) report
It is used for the survey of gamma interferon in the label-free electrochemistry aptamer sensor of graphene interface Controllable assembly using rolling ring amplifying technique
Fixed, the range of linearity for detecting gamma interferon is 0.1-0.7pM, and detection is limited to 0.065pM.
Graphene can influence the spin density and distribution of charges of carbon atom after N doping, and band structure has adjustment, led
Graphenic surface is caused to produce " avtive spot ", these avtive spots can directly participate in catalytic reaction, pass through N doping compound
Electro-chemical activity be improved significantly.MoS2There is unique and similar two-dimensional layer nanostructured, Liang Zhetong with graphene
The new heterogeneous interlayer structure of compound composition is crossed, the structure has new interaction, helps to obtain new, more excellent property
Energy.In addition, the complementarity in matching and electric property based on both on crystal structure and microscopic appearance, passes through both
Novel nanocomposite materials prepared by Material cladding can farthest show cooperative effect therebetween.
Although prior art has been disclosed for a variety of electrochemistry aptamer sensors and corresponding method of detection, for γ-dry
The detection of element is disturbed, its sensitivity how is improved, is still the focus of attention of those skilled in the art.
The content of the invention
For technical problem present in prior art, the present invention provides one kind and is based on three-dimensional nitrogen-doped graphene/curing
Fit electrode of gamma interferon of molybdenum and preparation method and application.
In order to solve problem above, the technical scheme is that：
A kind of preparation method of the fit electrode of interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide, including following step
Suddenly：
1) three-dimensional nitrogen-doped graphene/molybdenum disulfide composite (3D G-N/MoS2) preparation；
2) horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite (HRP/3D G-N/MoS2) system
It is standby
Weigh appropriate three-dimensional nitrogen-doped graphene/molybdenum disulfide composite to add in secondary water, ultrasonic resonance is for a period of time
Afterwards, horseradish peroxidase is added, ice-water bath stirring, it is multiple to obtain horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide
Condensation material, after filtering, horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is dispersed in water preservation；
3) horseradish peroxidase/aptamer modified electrode (ssDNA/ of three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
HRP/3D G-N/MoS2/ GCE) preparation
After bare glassy carbon electrode pretreatment, by the sulphur of the horseradish peroxidase obtained in step 2)/three-dimensional nitrogen-doped graphene/bis-
Change molybdenum composite material drop coating in glass-carbon electrode (GCE) surface, obtained horseradish peroxidase/three-dimensional nitrogen-doped graphene/curing
Molybdenum/naked glass carbon (HRP/3D G-N/MoS2/ GCE) modified electrode；
By the HRP/3D G-N/MoS of preparation2/ GCE modified electrodes are immersed in gamma interferon fit (ssDNA) and trained
Educate, enclosed-electrode, the aptamer modified electrode of gamma interferon electrochemistry (ssDNA/HRP/3D G-N/MoS are made2/GCE)。
Preferably, in step 2), three-dimensional nitrogen-doped graphene/molybdenum disulfide composite, horseradish peroxidase and water adds
The weight ratio entered is：1-3:4-12:1-3.
Preferably, in step 2), time of ultrasonic resonance is 1-3h, and ultrasonic resonance causes the sulphur of three-dimensional nitrogen-doped graphene/bis-
Change molybdenum to be uniformly dispersed in water.
Preferably, in step 2), the concentration of the horseradish peroxidase is 2-6mg/mL.Preferably, in step 2), ice
The time of stirring in water bath is 4-10h.
Preferably, in step 2), the preservation of horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
Temperature is 4 DEG C；The concentration that horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is dispersed in water is 8-
12mg/mL。
Preferably, in step 3), horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is added drop-wise to
The volume of electrode surface is 5-9 μ L.
Preferably, in step 3), the concentration of the ssDNA solution is 0.08-0.12mM；
Or the time that electrode soaks in ssDNA solution is 1-3h.
Preferably, in step 3), the method for enclosed-electrode is：SsDNA electrode will be soaked in 0.8-1.5mM 6- mercaptos
Base -1- own alcohol solution for soaking 0.5-1.5h, avoid light place 1h, electrode is cleaned with secondary water and cleaning fluid again, and obtained γ -
Interferon electrochemistry aptamer sensor, the electrode closed.
It is further preferred that the method for cleaning, comprises the following steps：Successively with 100mM phosphate buffer solutions (PBS),
Secondary water and 100mM PBSs.
Preferably, in step 3), the preprocess method of bare glassy carbon electrode, comprise the following steps：Use Al2O3Powder is by electrode table
Face is polished into minute surface, after cleaning, the ultrasonic 30s in secondary water and ethanol solution in succession, by the electrode cleaned up 0.5M's
Cyclic voltammetric is to stabilization in sulfuric acid solution, and after secondary water is rinsed well, room temperature is dried.
The horseradish peroxidase that the above method is prepared/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is fit
Modified electrode.
Above-mentioned horseradish peroxidase/three-dimensional nitrogen-doped graphene/aptamer modified electrode of molybdenum disulfide composite is detecting
Application in gamma interferon.
Above-mentioned horseradish peroxidase/aptamer modified electrode pair the γ of three-dimensional nitrogen-doped graphene/molybdenum disulfide composite-dry
The detection method of element is disturbed, is comprised the following steps：
1) the gamma interferon standard liquid of various concentrations is prepared；
2) by the horseradish peroxidase of preparation/three-dimensional nitrogen-doped graphene/aptamer modified electrode of molybdenum disulfide composite
It is immersed in the gamma interferon solution prepared in step 1), 0.8-1.2h is stood at 36-38 DEG C, after cleaning, the work that is assembled
Electrode；
3) be connected to by reference electrode, to electrode and working electrode on electrochemical workstation, in a cell add PBS,
Methylene blue (MB) and H2O2, then with square wave voltammetry detect working electrode current-responsive, according to the current-responsive of gained with
Relation between the concentration of gamma interferon standard liquid, drawing curve；
4) detection of actual sample：Actual sample is detected, according to the current-responsive measured and the work in step 3)
Make curve, draw the gamma interferon concentration of actual sample.
Preferably, in step 3), PBS concentration is 100mM in electrolytic cell, and MB concentration is 0.1mM, H2O2Concentration be
2mM。
Further, the invention also discloses the renovation process of above-mentioned electrochemistry aptamer sensor, after specially detecting, pass
Sensor is regenerated by cultivating 2h method in 1mM gamma interferons aptamer solutions (ssDNA).After cleaning, the electrode of regeneration can be with
For detecting next time.
The principle of the present invention is as follows：
The present invention utilize intermolecular electrostatic attraction, be prepared for gamma interferon it is fit/horseradish peroxidase/N doping
The aptamer modified electrode of graphene/molybdenum disulfide (ssDNA/HRP/3D G-N/MoS2/GCE).Nitrogen-doped graphene/molybdenum disulfide
(3D G-N/MoS2) compound is negatively charged in the solution, horseradish peroxidase (HRP) is positively charged in the solution, 3D G-
N/MoS2It is multiple that horseradish peroxidase/nitrogen-doped graphene/molybdenum disulfide is combined into by electrostatic attraction active force between HRP
Compound (HRP/3D G-N/MoS2), the material possesses good biocompatibility.SsDNA is intermolecular by electrostatic attraction and π-π
Active force and horseradish peroxidase/nitrogen-doped graphene/molybdenum disulfide modified electrode (/HRP/3DG-N/MoS2/ GCE) electrode
Surface combines, and forms ssDNA/HRP/3DG-N/MoS2The aptamer modified electrodes of/GCE.In the presence of gamma interferon, electricity
The gamma interferon on pole surface is fit to be combined with gamma interferon so that gamma interferon is fit to leave electrode surface, and what is exposed is peppery
Root peroxidase HRP is in MB-H2O2Electrochemical signals are produced in system, so as to realize the measure to gamma interferon.
The present invention advantageous effects be：
Class graphite ene compound crystal of molybdenum disulfide is made up of a metal molybdenum layer and two sulphur layers, forms two sulphur layer folders
The interlayer structure of metal molybdenum layer, and is formed by Van der Waals force accumulation, each molybdenum atom is surrounded by six sulphur atoms, in three
Angle prism-shaped, expose many MoS2Faceted pebble, strong adsorption force.Because graphene has big specific surface area, good electric conductivity
And chemical stability, MoS can be used as2Carrier, can effectively improve MoS2Agglomeration traits, give full play to both in crystal knot
The complementarity and its cooperative effect in matching and electric property on structure and microscopic appearance.
Three-dimensional nitrogen-doped graphene/molybdenum disulfide composite prepared by this method, there is big specific surface area, good lead
Electrically and chemical stability, big specific surface area can increase transmission rate and the measure sensitivity of electronics；Graphene is mixed through nitrogen
The spin density and distribution of charges of carbon atom can be influenceed after miscellaneous, band structure has adjustment, causes graphenic surface to produce " activity
Site ", these avtive spots can directly participate in catalytic reaction, and the electro-chemical activity of N doping compound is improved significantly,
So as to can further improve the conductive capability of modified electrode and sensitivity.
The present invention is based on nitrogen-doped graphene/molybendum disulfide complexes (3D G-N/MoS2) composite combines fit knowledge
Other technology and HRP-MB-H2O2System, make sensor that there is high sensitivity and selectivity.Use sensor of the present invention
Detected, the detectable concentration range of linearity of gamma interferon is 10-14-10-8M, lowest detection are limited to 3.3 × 10-15M。
Brief description of the drawings
Fig. 1 is the schematic diagram of preparation and the detection of aptamer sensor of the present invention；
Fig. 2 is the scanning electron microscope diagram of three-dimensional nitrogen-doped graphene/molybdenum disulfide composite；
HRP, ssDNA, 3D G-N/MoS in Fig. 3 present invention2And ssDNA/HRP/3DG-N/MoS2UV, visible light light splitting light
Spectrogram；
The testing result of Fig. 4 interferon：A) for various concentrations gamma interferon detection SWV curves, B) for peak current with
The linear relationship of the negative logarithm of gamma interferon concentration.
Embodiment
With reference to specific embodiments and the drawings, the invention will be further described.
Embodiment 1
Test reagent
Gamma interferon is fit (ssDNA)：5’-GGGGTTGGTTGTGTTGGGTGTTGTGTC CAA CCC C-3’；Interference
The nucleotide sequence that element is fit is purchased from raw work biology Co., Ltd with horseradish peroxidase (HRP).
Phosphate buffer solution (PBS), 6- sulfydryl -1- hexanols, methylene blue (MB), hydrogen peroxide (H2O2) it is purchased from Shanghai
AIBI chemicals Co., Ltd.Graphite powder, thioacetamide, calcium chloride, oxalic acid, sodium molybdate (Na2MoO4·2H2) and salt O
Acid, purchased from Solution on Chemical Reagents in Shanghai company.
Every other reagent is that analysis is pure.
Test apparatus
CHI 660C electrochemical workstations (Shanghai Chen Hua Instrument Ltd.), (Shanghai section leads ultrasound to ultrasonic cleaner
Instrument Ltd.), assay balance (Beijing Sai Duolisi balances Co., Ltd)；Reference electrode, auxiliary in three-electrode system
Electrode and working electrode are respectively saturated calomel electrode (SCE), platinum electrode and modified electrode.
The current potential that the present invention is previously mentioned is relative to SCE electrode potential, and experiment condition is all room temperature.
The preparation of the fit electrode of gamma interferon of three-dimensional nitrogen-doped graphene/molybdenum disulfide
The preparation method of the fit electrode of gamma interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide, including following step
Suddenly：
1) preparation of three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
Graphene oxide is prepared using improved Hummer methods.Three-dimensional nitrating is prepared using patent (201510289262.9)
Graphene/molybdenum disulfide composite (3D G-N/MoS2), taking 100mg GO to be scattered in the 100mL aqueous solution, (mass volume ratio is
1:1) in, 1.0g calcium chloride and 0.82g oxalic acid powder are added after 30min is stirred by ultrasonic, at the same by 1.0g thioacetamides and
0.32g sodium molybdates are dissolved in 50ml secondary waters, ultrasonic 30min, two kinds of solution are mixed, ultrasonic 2h.It is transferred to 1000W electromagnetism
When 80 DEG C are heated on stove, NaOH (0.5M) is added dropwise dropwise and adjusts the pH of solution to 10, while continue to heat, after reacting 8h,
Take precipitation, with secondary water washing for several times after, be dried in vacuo 8h at 60 DEG C, it is compound to produce nitrogen-doped graphene/molybdenum disulfide/calcium oxalate
Thing.Obtained nitrogen-doped graphene/molybdenum disulfide/calcium oxalate is dissolved in 5M aqueous hydrochloric acid solution, ultrasonic vibration 1h, repetition is washed
Wash three times, then neutrality is arrived for several times with secondary water washing, filter, be then dried in vacuo 8h under conditions of 60 DEG C, obtain three-dimensional and mix
Nitrogen graphene/molybdenum disulfide composite.
2) preparation of horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
Take 2mg three-dimensionals nitrogen-doped graphene/molybdenum disulfide composite, add 2mL ultra-pure waters, after ultrasonic resonance 1h, add
2mL HRP (4mg/mL), stir 6h in ice-water bath, filtering, add water cleaning three times, and horseradish peroxidase/three-dimensional is made and mixes
Nitrogen graphene/molybdenum disulfide composite (HRP/3DG-N/MoS2) be distributed in 1mL secondary water.Protected under conditions of 4 DEG C
Deposit.
3) prepared by the aptamer modified electrode of horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
(1) 0.3 μm of Al of bare glassy carbon electrode2O3Powder is polished into minute surface, after secondary water cleaning, in succession in secondary water and second
Ultrasonic 30s in alcoholic solution；By the electrode cleaned up, cyclic voltammetric to stabilization, secondary water is rinsed dry in 0.5M sulfuric acid solution
After net, room temperature is dried.
(2) 7.00 μ L HRP/3D G-N/MoS are taken2Drop coating is dried in electrode surface, lucifuge low temperature, and HRP/3D G- are made
N/MoS2/GCE。
(3) electrode by (2) processing is immersed in into ssDNA (0.1mM) to cultivate, room temperature 2h, cleaning.Containing 50 μ L 1mM
1h in MCH (6- sulfydryl -1- hexanols), with enclosed-electrode, (100mM PBS, pH7.4 are cleaned afterwards；Secondary water is cleaned；100mM
PBS, pH7.4), horseradish peroxidase/three-dimensional nitrogen-doped graphene/aptamer modified electrode of molybdenum disulfide composite is made
(ssDNA/HRP/3DG-N/MoS2/GCE)。
4) the aptamer modified electrode pair γ-interference of horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
The measure of element
(1) preparation of the gamma interferon detection solution containing various concentrations：Gamma interferon is in solution (100mM PBS, pH
7.4,100mM NaCl, 1mM MgCl2) in be configured to concentration be respectively 10-14-10-9M solution (100 μ L), is stored at 4 DEG C
It is standby；
(2) the aptamer modified electrode prepared is immersed in the gamma interferon solution of 100 μ L concentration knowns, and 1h is stood at 37 DEG C,
Cleaned afterwards with secondary water and cleaning fluid (100mM PBS, pH 7.4), the working electrode assembled；
(3) it is connected to by reference electrode, to electrode and working electrode on electrochemical workstation, adds 10mL in a cell
Solution (100mM PBS, pH 7.4,0.1mM MB, 2mM H2O2), then detect aptamer modified electrode with square wave voltammetry
Current-responsive, according to gained current-responsive (Δ I=I-I0, I0It is HRP-MB-H respectively with I2O2It is aptamer modified in gamma interferon
Peak current before and after electrode and gamma interferon effect) relation between the concentration of standard solution of gamma interferon, paint
Working curve processed.
Cleaning Principle schematic diagram
The Cleaning Principle of gamma interferon is as shown in Figure 1：Nitrogen-doped graphene/molybdenum disulfide (3D G-N/MoS2) compound
Solution is negatively charged, and horseradish peroxidase (HRP) solution is positively charged, 3D G-N/MoS2Pass through electrostatic attraction between HRP
Active force is combined into HRP/3D G-N/MoS2Compound, ssDNA can be intermolecular by electrostatic attraction and π-π in electrode surface
Active force, it is incorporated in HRP/3D G-N/MoS2Electrode surface, form ssDNA/HRP/3D G-N/MoS2/ GCE modified electrodes.
In the presence of gamma interferon, gamma interferon and the fit combination of gamma interferon, gamma interferon is fit to leave electrode surface,
Electrode is containing MB-H2O2Electrochemical signals change in solution, increase with the increase electrochemical signals of gamma interferon concentration,
So as to realize the electrochemical gaging of aptamer modified electrode pair gamma interferon indirectly.
Experimental result
1st, characterize：The scanning electron microscope diagram of three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
As shown in Fig. 2 three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is cellular three-dimensional net structure, have big
Specific surface area, good electric conductivity and chemical stability, big specific surface area can increase the transmission rate and measure of electronics
Sensitivity；Graphene can influence the spin density and distribution of charges of carbon atom after N doping, and band structure has adjustment, caused
Graphenic surface produces " avtive spot ", and these avtive spots can directly participate in catalytic reaction, pass through N doping compound
Electro-chemical activity is improved significantly, so as to can further improve the conductive capability of fit electrode and sensitivity.
As shown in Fig. 3 ultraviolet-visible absorption spectroscopy figures, HRP absorption peak position in 402nm, compound at 402nm still
This peak so be present, illustrate HRP and 3D-GN/MoS2It is combined by electrostatic interaction；Gamma interferon is fit, and the ultraviolet of ssDNA can
See absworption peak in 260nm, ssDNA and HRP/3D-GN/MoS2SsDNA peak and 3D-GN/MoS after compound2Peak coexist and occur
It is overlapping.
3rd, the detection of gamma interferon：As shown in figure 4, the change of the SWV of gamma interferon electrochemical signals reflect γ-
The concentration of interferon, detection range are about 10-14-10-8M, test limit are about 3.3 × 10-15M, realize the Gao Ling of gamma interferon
Sensitivity detects.
4th, the gamma interferon measure of actual sample：Gamma interferon in human serum sample is determined, as a result seen
Table 1.The rate of recovery of gamma interferon is respectively 92.80-102.2%.In experiment, each sample Parallel testing five times, relative mark
Quasi- deviation is less than 5%.Result above shows that this method has good applicability.
Detection of the table 1 in 300 times of human serum is diluted to gamma interferon
Above-mentioned although the embodiment of the present invention is described with reference to accompanying drawing, not to invention protection domain
Limitation, one of ordinary skill in the art should be understood that on the basis of technical scheme those skilled in the art are not required to
Pay various modifications that creative work can make or deformation is still within the scope of the present invention.
Claims (10)
1. a kind of preparation method of the fit electrode of gamma interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide, its feature exist
In：Comprise the following steps：
1) preparation of three-dimensional nitrogen-doped graphene/molybdenum disulfide composite；
2) preparation of horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite
Appropriate three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is weighed to add in secondary water, ultrasonic resonance for a period of time after, add
Enter horseradish peroxidase, ice-water bath stirring, obtain horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite wood
Material, after filtering, preservation is dispersed in water by horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite；
3) preparation of horseradish peroxidase/three-dimensional nitrogen-doped graphene/aptamer modified electrode of molybdenum disulfide composite
It is after pretreatment of glassy carbon electrode, the horseradish peroxidase obtained in step 2)/three-dimensional nitrogen-doped graphene/molybdenum disulfide is multiple
Condensation material drop coating is repaiied in glassy carbon electrode surface, obtained horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide/naked glass carbon
Adorn electrode；
The horseradish peroxidase of preparation/three-dimensional nitrogen-doped graphene/molybdenum disulfide/naked glass carbon modified electrode is immersed in γ-interference
Cultivated in plain fit ssDNA, enclosed-electrode, the aptamer modified electrode of gamma interferon electrochemistry is made.
2. preparation method according to claim 1, it is characterised in that：In step 2), three-dimensional nitrogen-doped graphene/molybdenum disulfide
The weight ratio that composite, horseradish peroxidase and water add is：1-3:4-12:1-3；
Or in step 2), the time of ultrasonic resonance is 1-3h；
Or in step 2), the concentration of the horseradish peroxidase is 2-6mg/mL；
Or in step 2), the time of ice-water bath stirring is 4-10h；
Or in step 2), the storage temperature of horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is 4 DEG C；
The concentration that horseradish peroxidase/three-dimensional nitrogen-doped graphene/molybdenum disulfide composite is dispersed in water is 8-12mg/mL.
3. preparation method according to claim 1, it is characterised in that：In step 3), horseradish peroxidase/three-dimensional nitrating
The volume that graphene/molybdenum disulfide composite is added drop-wise to electrode surface is 5-9 μ L；
Or in step 3), the concentration of the ssDNA solution is 0.08-0.12mM；
Or the time that electrode soaks in ssDNA solution is 1-3h.
4. preparation method according to claim 1, it is characterised in that：In step 3), the method for enclosed-electrode is：Will immersion
SsDNA electrode is crossed in 0.8-1.5mM 6- sulfydryls -1- own alcohol solution for soaking 0.5-1.5h, avoid light place 1h, by electrode again
It is secondary to use secondary water and PBS, the fit electrode of gamma interferon electrochemistry is made.
5. preparation method according to claim 4, it is characterised in that：The method of cleaning, comprises the following steps：Use successively
100mM PBS, secondary water and 100mM PBSs.
6. preparation method according to claim 1, it is characterised in that：In step 3), the preprocess method of bare glassy carbon electrode,
Comprise the following steps：Use Al2O3Electrode surface is polished into minute surface by powder, in succession ultrasonic in secondary water and ethanol solution after cleaning
30s, by the electrode cleaned up, cyclic voltammetric is to stabilization in 0.5M sulfuric acid solution, and after secondary water is rinsed well, room temperature is dried in the air
It is dry.
7. the horseradish peroxidase that any methods describeds of claim 1-6 are prepared/three-dimensional nitrogen-doped graphene/molybdenum disulfide
The aptamer modified electrode of composite.
8. horseradish peroxidase described in the claim 7/three-dimensional nitrogen-doped graphene/aptamer modified electrode of molybdenum disulfide composite
Application in gamma interferon is detected.
9. horseradish peroxidase described in the claim 7/three-dimensional nitrogen-doped graphene/aptamer modified electrode of molybdenum disulfide composite
To the detection method of gamma interferon, comprise the following steps：
1) the interferon standard liquid of various concentrations is prepared；
2) horseradish peroxidase of preparation/three-dimensional nitrogen-doped graphene/aptamer modified electrode of molybdenum disulfide composite is immersed in
In the solution of the gamma interferon prepared in step 1), 0.8-1.2h is stood at 36-38 DEG C, after cleaning, the work assembled is electric
Pole；
3) drawing curve；
4) detection of actual sample.
10. detection method according to claim 9, it is characterised in that：In step 3), PBS concentration is in electrolytic cell
100mM, the concentration of methylene blue is 0.1mM, H2O2Concentration be 2mM.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610137834.6A CN105784819B (en) | 2016-03-11 | 2016-03-11 | A kind of fit electrode of interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide and preparation method and application |
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| CN201610137834.6A CN105784819B (en) | 2016-03-11 | 2016-03-11 | A kind of fit electrode of interferon based on three-dimensional nitrogen-doped graphene/molybdenum disulfide and preparation method and application |
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| CN103940872A (en) * | 2014-04-30 | 2014-07-23 | 青岛大学 | Preparation method and application of electrochemical sensor capable of simultaneously detecting two acute leukemia markers |
| CN104090116A (en) * | 2014-07-21 | 2014-10-08 | 扬州大学 | Preparation method for bovine gamma interferon impedance type immunosensor based on zinc oxide nano-materials |
| CN104835654A (en) * | 2015-05-29 | 2015-08-12 | 青岛大学 | 3D nitrogen-doped graphene/molybdenum disulfide compound and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103940872A (en) * | 2014-04-30 | 2014-07-23 | 青岛大学 | Preparation method and application of electrochemical sensor capable of simultaneously detecting two acute leukemia markers |
| CN104090116A (en) * | 2014-07-21 | 2014-10-08 | 扬州大学 | Preparation method for bovine gamma interferon impedance type immunosensor based on zinc oxide nano-materials |
| CN104835654A (en) * | 2015-05-29 | 2015-08-12 | 青岛大学 | 3D nitrogen-doped graphene/molybdenum disulfide compound and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| A highly sensitive label-free electrochemical aptasensor for interferon-gamma detection based on graphene controlled assembly and nuclease cleavage-assisted target recycling amplification;Genping Yan等;《BiosensorsandBioelectronics》;20130116;第44卷;第57-63页 * |
| Electrochemical immunosensor for interferon-γ based on disposable ITO detector and HRP-antibody-conjugated nano gold as signal tag;Yaru Zhang等;《Materials Science and Engineering C》;20151021;第59卷;第577–584页 * |
| Investigations of an electrochemical platform based on the layered MoS2–graphene and horseradish peroxidase nanocomposite for direct electrochemistry and electrocatalysis;HaiyanSong等;《Biosensors andBioelectronics》;20140111;第56卷;第137-143页 * |
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